Journal articles on the topic 'Budderoo National Park (N.S.W.)'

Based on a study of 1,100 specimens of the genus Agra (Coleoptera: Carabidae: Lebiini: Agrina) from Costa Rica, twenty-nine new species were discovered and are here validated and described: A. catie, n. sp. (type locality: Limón, Tortuguero National Park, Estacíon Cuatro Esquinas, sea level, LN 280000,590500); A. catbellae, n. sp. (type locality: Cartago, Turrialba, 600m, 09º53'N 083º38'W); A. dable, n. sp. (type locality: Heredia, Estacíon Magsasay, Parque Nacional Braulio Carrillo, 200m, LN 264600,531100); A. delgadoi, n. sp. (type locality: Cartago, Turrialba, CATIE, 600m, 09º53'N 083º38'W); A. fugax, n. sp. (type locality: Heredia, Estacíon La Selva, 10º27'N 083º59'W; A. giesberti, n. sp. (type locality: Cartago, 15km NE Turrialba, 10º00'N 083º30'W); A. granodeoro, n. sp. (type locality: Cartago, Turrialba, Chirripo, Grano de Oro, 1120m, LN 200250,595900); A. ichabod, n. sp. (type locality: Alajuela, Atenas, 9º58'N 084º23'W); A. jimwappes, n. sp. (type locality: Guanacaste, La Pacifica, 10º28'N 085º07'W); A. julie, n. sp. (type locality: Cartago, Turrialba, 600m, 09º53'N 083º38'W); A. katewinsletae, n. sp. (type locality: Puntarenas, Monteverde, 1380m, 10º50'N 085º37'W); A. liv, n. sp. (type locality: Puntarenas, Manual Antonio National Park, Quepos, 80m, 09°23'N 84°09'W); A. monteverde, n. sp. (type locality: Puntarenas, Monteverde, 1380m, 10º50'N 085º37'W); A. not, n. sp. (type locality: Puntarenas, Carara Biological Reserve, Estacíon Bonita, 50m, LN 194500,469850); A. notcatie, n. sp. (type locality: Limón, Tortuguero National Park, Estacíon Cuatro Esquinas, sea level, LN 280000,590500, November (R. Delgado)(INBio: CRI000-298655); A. pitilla, n. sp. (type locality: Guanacaste, Guanacaste National Park, Estacíon Pitilla, 9 km S Santa Cecilia, 700m, LN 330200,380200); A. phallica, n. sp. (type locality: Cartago, Tucurrique, 09º51'N 083º43'W); A. quesada, n. sp. (type locality: Limón, Manzanillo, RNFS Gandoca y Manzanillo, 0-10 sea level, LS 398100,610600); A. santarosa, n. sp. (type locality: Guanacaste, Santa Rosa National Park, 280m, 10º50'N 085º37'W); A. schwarzeneggeri, n. sp. (type locality: Cartago, Turrialba, 650m, 09º53'N 083º38'W); A. sirena, n. sp. (type locality: Heredia, Estacíon La Selva, 10º27'N 083º59'W); A. solanoi, n. sp. (type locality: ); A. solisi, n. sp. (type locality: Limón, Tortuguero National Park, Cerro Tortuguero, 119m, LN 285000,588000); A. turrialba, n. sp. (type locality: Cartago, Turrialba, 600m, 09º53'N 083º38'W);A. ubicki, n. sp. (type locality: Puntarenas, 3 km NE Golfito, 8º39'N 083º10'W); A. winnie, n. sp. (type locality: Guanacaste, Guanacaste National Park, Estacíon Santa Rosa, 800m, LN 313000,359800); A. zumbado, n. sp. (type locality: Guanacaste, Guanacaste National Park, Estacíon Patilla, 9 km S Santa Cecilia, 700m, LN 330200,380200); A. zuniga, n. sp. (type locality: Puntarenas, Manual Antonio National Park, Quepos, 80m, LS 370900,448800). Six additional species are recorded for the first time in Costa Rica: Agra castaneipes Bates, A. campana Erwin, A. fortuna Erwin, A. guatemalena Csiki, A. incisa Liebke, and A. rufiventris Bates. The presence of a Panamanian species, A. championi Bates, in Costa Rica, as noted by Max Liebke has been confirmed (Agra danjanzeni Erwin = A. championi Bates, new synonymy). Also included: Neotype designation for Agra pia Liebke 1940 and apparent rediscovery of this species in Costa Rica; Agra aurifera Liebke 1940 description translated from the German and reproduced here with comments.

Two new species of anilline ground beetles are described from the Appalachian Mountains of eastern United States. The description of Serranillus septentrionis n. sp. is based on specimens collected in montane areas of western Virginia (37°25.33’N, 79°45.43’W). This species extends the range of the genus approximately 200 km north of its closest known congeners, S. dunavani (Jeannel) and S. jeanneli Barr, and differs from them mainly in characters of the male genitalia. A key is provided that will allow separation of these three species without dissection. The description of Anillinus cherokee n. sp. is based on specimens collected in Great Smoky Mountains National Park and nearby areas of Nantahala National Forest, western North Carolina (35°21.33’N, 83°56.05’W). The species is externally similar to A. loweae Sokolov and Carlton and A. steevesi Barr, and differs from those species in characters of the male genitalia.

The Australian species of the water beetle genus Hydraena Kugelann, 1794, are revised, based on the study of 7,654 specimens. The 29 previously named species are redescribed, and 56 new species are described. The species are placed in 24 species groups. High resolution digital images of all primary types are presented (online version in color), and geographic distributions are mapped. Male genitalia, representative female terminal abdominal segments and representative spermathecae are illustrated. Australian Hydraena are typically found in sandy/gravelly stream margins, often in association with streamside litter; some species are primarily pond dwelling, a few species are humicolous, and one species may be subterranean. The areas of endemicity and species richness coincide quite closely with the Bassian, Torresian, and Timorian biogeographic subregions. Eleven species are shared between the Bassian and Torresian subregions, and twelve are shared between the Torresian and Timorian subregions. Only one species, H. impercepta Zwick, is known to be found in both Australia and Papua New Guinea. One Australian species, H. ambiflagellata, is also known from New Zealand. New species of Hydraena are: H. affirmata (Queensland, Palmerston National Park, Learmouth Creek), H. ambiosina (Queensland, 7 km NE of Tolga), H. antaria (New South Wales, Bruxner Flora Reserve), H. appetita (New South Wales, 14 km W Delagate), H. arcta (Western Australia, Synnot Creek), H. ascensa (Queensland, Rocky Creek, Kennedy Hwy.), H. athertonica (Queensland, Davies Creek), H. australula (Western Australia, Synnot Creek), H. bidefensa (New South Wales, Bruxner Flora Reserve), H. biimpressa (Queensland, 19.5 km ESE Mareeba), H. capacis (New South Wales, Unumgar State Forest, near Grevillia), H. capetribensis (Queensland, Cape Tribulation area), H. converga (Northern Territory, Roderick Creek, Gregory National Park), H. cubista (Western Australia, Mining Camp, Mitchell Plateau), H. cultrata (New South Wales, Bruxner Flora Reserve), H. cunninghamensis (Queensland, Main Range National Park, Cunningham's Gap, Gap Creek), H. darwini (Northern Territory, Darwin), H. deliquesca (Queensland, 5 km E Wallaman Falls), H. disparamera (Queensland, Cape Hillsborough), H. dorrigoensis (New South Wales, Dorrigo National Park, Rosewood Creek, upstream from Coachwood Falls), H. ferethula (Northern Territory, Cooper Creek, 19 km E by S of Mt. Borradaile), H. finniganensis (Queensland, Gap Creek, 5 km ESE Mt. Finnigan), H. forticollis (Western Australia, 4 km W of King Cascade), H. fundaequalis (Victoria, Simpson Creek, 12 km SW Orbost), H. fundata (Queensland, Hann Tableland, 13 km WNW Mareeba), H. hypipamee (Queensland, Mt. Hypipamee National Park, 14 km SW Malanda), H. inancala (Queensland, Girraween National Park, Bald Rock Creek at "Under-ground Creek"), H. innuda (Western Australia, Mitchell Plateau, 16 mi. N Amax Camp), H. intraangulata (Queensland, Leo Creek Mine, McIlwrath Range, E of Coen), H. invicta (New South Wales, Sydney), H. kakadu (Northern Territory, Kakadu National Park, Gubara), H. larsoni (Queensland, Windsor Tablelands), H. latisoror (Queensland, Lamington National Park, stream at head of Moran's Falls), H. luminicollis (Queensland, Lamington National Park, stream at head of Moran's Falls), H. metzeni (Queensland, 15 km NE Mareeba), H. millerorum (Victoria, Traralgon Creek, 0.2 km N 'Hogg Bridge', 5.0 km NNW Balook), H. miniretia (Queensland, Mt. Hypipamee National Park, 14 km SW Malanda), H. mitchellensis (Western Australia, 4 km SbyW Mining Camp, Mitchell Plateau), H. monteithi (Queensland, Thornton Peak, 11 km NE Daintree), H. parciplumea (Northern Territory, McArthur River, 80 km SW of Borroloola), H. porchi (Victoria, Kangaroo Creek on Springhill Rd., 5.8 km E Glenlyon), H. pugillista (Queensland, 7 km N Mt. Spurgeon), H. queenslandica (Queensland, Laceys Creek, 10 km SE El Arish), H. reticuloides (Queensland, 3 km ENE of Mt. Tozer), H. reticulositis (Western Australia, Mining Camp, Mitchell Plateau), H. revelovela (Northern Territory, Kakadu National Park, GungurulLookout), H. spinissima (Queensland, Main Range National Park, Cunningham's Gap, Gap Creek), H. storeyi (Queensland, Cow Bay, N of Daintree River), H. tenuisella (Queensland, 3 km W of Batavia Downs), H. tenuisoror (Australian Capital Territory, Wombat Creek, 6 km NE of Piccadilly Circus), H. textila (Queensland, Laceys Creek, 10 km SE El Arish), H. tridisca (Queensland, Mt. Hemmant), H. triloba (Queensland, Mulgrave River, Goldsborough Road Crossing), H. wattsi (Northern Territory, Holmes Jungle, 11 km NE by E of Darwin), H. weiri (Western Australia, 14 km SbyE Kalumburu Mission), H. zwicki (Queensland, Clacherty Road, via Julatten).

ABSTRACT The Aquificales are widespread in marine and terrestrial hydrothermal environments. Here, we report the complete and draft genome sequences of six new members of the Aquificales: two marine species, Persephonella marina strain EX-H1 and Hydrogenivirga strain 128-5-R1 (from the East Pacific Rise, 9°50.3′N, 104°17.5′W, and the Eastern Lau Spreading Center, 176°11.5′W, 20°45.8′S, respectively), and four terrestrial isolates, Sulfurihydrogenibium azorense strain Az-Fu1, Sulfurihydrogenibium yellowstonense strain SS-5, and Sulfurihydrogenibium strain Y03AOP1 (from Furnas, Azores, Portugal, and Calcite Springs and Obsidian Pool in Yellowstone National Park, United States, respectively), and the only thermoacidophilic isolate, Hydrogenobaculum strain Y04AAS1 (from a stream adjacent to Obsidian Pool). Significant differences among the different species exist that include nitrogen metabolism, hydrogen utilization, chemotaxis, and signal transduction, providing insights into their ecological niche adaptations.

<p>The Scalloped hammerhead shark, <em>Sphyrna lewini</em> is a coastal and pelagic circumglobal species that resides within coastal warm temperate and tropical seas. <em>Sphyrna lewini</em> exhibits strong intraspecific segregation: neonates and young-of-the-year spend the first part of life in coastal inshore waters (nursery grounds), while adults migrate offshore, returning to protected nursery habitats for mating and pupping. On December 3, 2014, at approximately 19:00 hr, four young-of-the-year <em>S. lewini</em> were caught with hand line in Wafer Bay, Isla del Coco, Costa Rica (5°32’42.4” N - 87°03’45.3” W). A total of three males (total length (TL): 73, 73, 76 cm) and one female (TL: 75 cm) were recorded. The presence of these individuals at Isla del Coco suggests that a pregnant female gave birth in or near Wafer Bay, which may be a nursery ground for <em>S. lewini</em>. We recommend further study to evaluate the presence and movements of young-of-the-year and juvenile <em>S. lewini</em> in Wafer Bay to determine if this was an isolated incident or if the bay is a nursery ground for <em>S. lewini</em>.</p><div> </div>

Relief and geological structure of Hutsulshchyna and Verkhovynskyi national natural parks (NNP), located in the south-eastern part of the Ukrainian Carpathians, present four geomorphological regions of the Ukrainian Carpathians. Hutsulshchyna NNP is located in Skybovi Carpathians and Precarpathian Upland. Verkhovynskyi NNP is located in Marmaroski Carpathians and Polonynsko-Chornohirski Carpathians. The analysis of the morphostructure and morphosculpture of national natural parks is carried out taking into account the longitudinal (N-W – S-E) and transverse divisions of the Ukrainian Carpathians. The longitudinal division is associated with morphostructures of higher orders, such as the second and third, and the transverse is associated with the fourth and fifth orders of morphostructure. In the analysis of morphosculpture of NNPs, the types which are characteristic of the Carpathian Flysch and Сrystalline Carpathians are allocated. Mountain ranges and ridges such as Sloboda-Rungurska, Pokuttia med-mountains, Hryniava-Losova and Chyvchyn mountains are characterized by an asymmetrical structure – steep northeastern slopes and declivous southwestern slopes. The relic morphosculpture is represented by: 1) fragments of denudation surfaces of different ages such as Polonynska within Verkhovynskyi NPP as well as Karmaturska (analogue of Pidbeskid) within Hutsulshchyna NPP, and riparian; 2) extra and ancient glacial within Verkhovynskyi NPP and extra glacial within Hutsulshchyna NPP; 3) areas of ancient longitudinal valleys. The presence of relict ("dead") river valleys is characteristic of the premountain part of Hutsulshchyna NNP. Modern morphodynamic processes represent by height (tier) differentiation. In the tiers of strongly dissected mеd-mountain and low-mountain relief, the processes of planar erosion, deflux, and linear erosion play an important role in the modelling of the relief. The lower tier of the terraced and non-terraced bottoms of the valleys are associated with the processes of leaching and erosion as well as a significant accumulation of erosion products and mudflows. Among gravitational processes and block motions, stabilized and active displacements are the most recorded. Key words: National natural park; Ukrainian Carpathians; relief; morphostructure; morphosculpture.

It has been challenging to synthesize p-type SnOx (1≤x<2) and engineer the electrical properties such as carrier density and mobility due to the narrow processing window and the localized oxygen 2p orbitals near the valence band. We recently reported on the processing of p-type SnOx and an oxide-based p-n heterostructures, demonstrating high on/off rectification ratio (>103), small turn-on voltage (<0.5 V), and low saturation current (~1×10-10 A)1. In order to further understand the p-type oxide and engineer the properties for various electronic device applications, it is important to identify (or establish) the dominating doping and transport mechanisms. The low dopability in p-type SnOx, of which the causation is also closely related to the narrow processing window, needs to be mitigated so that the electrical properties of the material are to be adequately engineered 2, 3. Herein, we report on the multifunctional encapsulation of p-SnOx to limit the surface adsorption of oxygen and selectively permeate hydrogen into the p-SnOx channel for thin film transistor (TFT) applications. Time-of-flight secondary ion mass spectrometry measurements identified that ultra-thin SiO2 as a multifunctional encapsulation layer effectively suppressed the oxygen adsorption on the back channel surface of p-SnOx and augmented hydrogen density across the entire thickness of the channel. Encapsulated p-SnOx-based TFTs demonstrated much-enhanced channel conductance modulation in response to the gate bias applied, featuring higher on-state current and lower off-state current. The relevance between the TFT performance and the effects of oxygen suppression and hydrogen permeation is discussed in regard to the intrinsic and extrinsic doping mechanisms. These results are supported by density-functional-theory calculations. Acknowledgement This work was supported by the U.S. National Science Foundation (NSF) Award No. ECCS-1931088. S.L. and H.W.S. acknowledge the support from the Improvement of Measurement Standards and Technology for Mechanical Metrology (Grant No. 20011028) by KRISS. K.N. was supported by Basic Science Research Program (NRF-2021R11A1A01051246) through the NRF Korea funded by the Ministry of Education. References Lee, D. H.; Park, H.; Clevenger, M.; Kim, H.; Kim, C. S.; Liu, M.; Kim, G.; Song, H. W.; No, K.; Kim, S. Y.; Ko, D.-K.; Lucietto, A.; Park, H.; Lee, S., High-Performance Oxide-Based p–n Heterojunctions Integrating p-SnOx and n-InGaZnO. ACS Applied Materials & Interfaces 2021, 13 (46), 55676-55686. Hautier, G.; Miglio, A.; Ceder, G.; Rignanese, G.-M.; Gonze, X., Identification and design principles of low hole effective mass p-type transparent conducting oxides. Nat Commun 2013, 4. Yim, K.; Youn, Y.; Lee, M.; Yoo, D.; Lee, J.; Cho, S. H.; Han, S., Computational discovery of p-type transparent oxide semiconductors using hydrogen descriptor. npj Computational Materials 2018, 4 (1), 17. Figure 1

In the stydy an analysis of the geological structure and relief of Uzhansky, “Skolivsky Beskydy”, and “Boykivshchyna” national natural parks (NNP), located in the north-western part of the Ukrainian Carpathians, is presented. Uzhansky NNP is located within the Polonynsko-Chornohirska and Vododilno-Verkhovyna geomorphological regions, “Skolivski Beskydy” NNP is situated in the Skibovy Carpathians, and the newly created “Boykivshchyna” NNP is located within the Vododilno-Verkhovyna and Skybovi Carpathians. The analysis of the morphostructure and morphosculpture of national parks is carried out taking into account the longitudinal (N-W–S-E) and transverse divisions of the Ukrainian Carpathians. The longitudinal division is associated with higher morphostructures of higher orders – the second and third, with the transverse is associated with the fourth and fifth morphostructures. In the analysis of morphosculpture of national parks, the types which are characteristic of the Carpathian Flysch belt are allocated. All mountain ranges and ridges are characterized by an asymmetrical structure – steep northeastern slopes and declivous southwestern slopes. The relic morphosculpture is represented by: 1) fragments of denudation surfaces of different ages such as Beskid, Pidbeskid, and riparian; 2) extra glacial and firn glaciations; 3) areas of ancient longitudinal valleys. Inherited morphosculpture is represented by river valleys with a complex of terraces of different ages. Modern morphodynamic processes represent by height (tier) differentiation. In the tiers of strongly dissected mid-mountain and low-mountain relief, the processes of planar erosion, deflux, and linear erosion play an important role in the modeling of the relief. The lower tier of the terraced and non-terraced bottoms of the valleys are associated with the processes of leaching and erosion as well as a significant accumulation of erosion products and mudflows. Among gravitational processes and block motions, stabilized and active displacements are the most recorded. Keywords: National natural park; Ukrainian Carpathians; relief; morphostructure; morphosculpture.

The hummingbird-visited plant community located on the open-habitat mountaintop of the Espinhaço Range was studied for two years (from August 2007 to July 2009) in Serra do Cipó National Park, Southeastern Brazil (19° 15′ S and 43° 31′ W). The floral characteristics and flowering period of the hummingbird-visited plants was monthly recorded along trails located in three vegetation types: (1) typical campos rupestres (TCR), (2) open fields (OPF), and (3) capões de mata(CAM). Hummingbird visitation was observed in 51 plant species, 22 ornithophilous and 29 non-ornithophilous species. The TCR showed the greatest number of species visited (N = 38), followed by the OPF (N = 18) and CAM (N = 17). Six species of hummingbirds were recorded visiting flowers: Augastes scutatus, Campylopterus largipennis, Colibri serrirostris, Chlorostilbon lucidus, Eupetomena macroura and Phaethornis pretrei. This study demonstrates that the species richness and the number of ornithophilous species visited by the hummingbirds at the study site are more similar to hummingbird-plant communities of the Atlantic Forest than to those of the Cerrado communities and other Brazilian highland open-habitat communities. The plant families most visited by hummingbirds were Bromeliaceae and Asteraceae. Although the Asteraceae family is rarely used as a food resource for hummingbirds in other high and lowland communities, in the study site this family is used mainly by the endemic hummingbird Augastes scutatus. We found a large overlap of flowering throughout the year among the species visited by the hummingbirds. Thus, the nectar availability supports these resident hummingbirds. The present study also showed that the studied hummingbird-plant community is composed of many species endemic to the campos rupestres of the Espinhaço Range, some of which are considered to be in danger of extinction, thus constituting a unique and threatened community. Thus, understanding hummingbird-plant pollination dynamics becomes fundamental to the conservation of the campos rupestres.

The studies were carried out in 12 beech stands, growing on mountain fresh forest site in Magurski National Park. Stands differed in respect of exposition (northern and southern) and age - lower age classes (21-60 years old) and higher age classes (81-120 years old). This paper presents evaluation of frequency of disease symptoms occurrence in the crowns and on the trunks of beeches. The presence and the place of occurrence of fungus <i>Ascodichaena rugosa</i> and insects <i>Cryptococcus fagisuga</i> and <i>Ectoedemia liebwerdella</i>, which could be connected with beech bark necrosis on the trunks were noted down. In each stand 50 trees, growing side by side in center of the stand, were analyzed. Among 600 analyzed trees of <i>Fagus sylvatica</i>, 2.6% had a dead top. Crown thinning were present on 51.8% of trees. It appeared frequently in higher age class stands and didn't cover more than ¼ of the crown. Moreover, in crowns of 4.1% of trees the leaf chlorosis appeared. The most frequent symptom on trunks of beeches were local necroses (98% of trees), that were sometimes connected with slime fluxing (3.3%). Necroses were the most number in stands growing on the southern exposition and on south parts of the trunks. <i>Cryptococcus fagisuga</i> appeared mostly in high age class stands. Intensity of its appearance was similar on the trees growing on the southern and northern exposition (respectively, 48.3% and 53.3%). Particular parts of the trunks were colonized with similar frequency by <i>C. fagisuga</i> irrespective of cardinal points (N-29.3%; S-21.7%; W-22.7%; E-26.2%) and number of necroses on the trunks. <i>Ascodichaena rugosa</i> appeared on 8.6% of trees, more frequently on their north parts of the trunks (7.3%). In the places colonized by fungi <i>A. rugosa</i>, insect <i>C. fagisuga</i> was absent. Occurrence of larvae of <i>Ectoedemia liebwerdella</i> was mostly found on trunks of beeches growing on the southern exposition (43.0%), on 35.3% of trees they colonized south parts of trunks. There wasn't observed any connection between the occurrence of these larvae and symptoms of local necroses.

<p>This is Part II of a two-part review about climate and climate variability focused on the Eastern Tropical Pacific (ETP) and the Caribbean Sea (CS). Both parts are aimed at providing oceanographers, marine biologists, and other ocean scientists, a guiding base for ocean-atmosphere interaction processes affecting the CS, the ETP, and the waters of Isla del Coco. Isla del Coco National Park is a Costa Rican World Heritage site. Part I analyzed the mean fields for both basins and a larger region covering 25º S - 35º N, 20º W - 130º W. Here we focus on a smaller area (65º W - 95º W, 0º - 20º N), as a complement to Part 1. Incoming solar radiation and surface energy fluxes reveal the complex nature of the ETP and CS for convective activity and precipitation on seasonal and intraseasonal time scales. Both regions are relevant as sources of evaporation and the associated moisture transport processes. The American Monsoon System influences the climate and climate variability of the ETP and CS, however, the precise way systems affect regional precipitation and transport of moisture, within the Intra Americas Sea (IAS) are not clear. Although the Caribbean Low-Level Jet (CLLJ) is known to act as a conveyor belt for moisture transport, intraseasonal and seasonal modes of the CLLJ and their interactions with other IAS systems, have to be further investigated. Trans-isthmic jets, exert a variable seasonal wind stress force over the ocean surface co-generating regions of great marine productivity. Isolated convection, the seasonal migration of the Intertropical Convergence Zone, the hurricane season, the Mid-Summer Drought, the seasonal and intraseasonal behavior of low-level jets and their interactions with transients, and the southward incursion of cold fronts contribute to regional seasonal precipitation. Many large-scale systems, such as El Niño-Southern Oscillation, the Atlantic Multidecadal Oscillation and the Madden-Julian Oscillation (MJO, also influence the variability of precipitation by modulating regional features associated with convection and precipitation. Monthly tropical storm (TS) activity in the CS and ETP basins is restricted to the period May-November, with very few cases in December. The CS presents TS peak activity during August, as well as for the number of hurricanes and major hurricanes, in contrast to the ETP that shows the same features during September.</p><div> </div>

Aim of the study: to analyze, within a Pinus contorta plantation, the effects of artificially created small rectangular and small medium circular canopy gaps on: i) photosynthetic active radiation (PAR), and soil temperature and moisture, and ii) survival and growth of planted Austrocedrus chilensis and Nothofagus dombeyi seedlings, species which formerly composed the natural forest of the area.Study area: A 2 ha stand of a Pinus contorta stand in Los Alerces National Park, Argentina (42°43’S, 71°43’W, 490 m.a.s.l.). Material and methods: The Pinus contorta stand was 25 yr old, 22 m height and 26 cm DBH, presenting 1000 trees ha-1 of density and 53 m2 ha-1 of basal area. In 2009, rectangular and circular gaps were created within the stand and then seedlings were planted. During two growing seasons (2010-2011 and 2011-2012), PAR, soil temperature and moisture were measured in gaps and understory (control), and seedling survival and growth in gaps.Main results: During both seasons, soil temperature did not differ among gaps and control, whereas PAR and soil moisture were lower in control than in gaps. Seedling survival was high in all gaps regardless of species and season. Seedlings showed higher diameter growth in rectangular than in circular gaps.Research highlights: Austrocedrus chilensis and N. dombeyi seedlings survival is high and their growth slightly affected, when planted in differently-sized canopy gaps within a Pinus contorta plantation in Patagonia. However, other gap sizes and stand densities should be tested before recommending which one shows better results for reconverting monocultures into former native forests.Keywords: planted seedlings; gap structure; photosynthetic active radiation; soil temperature; soil moisture.Abbreviations used: PAR (Photosynthetic Active Radiation); DBH (Diameter at Breast Height); INTA (Argentinean Institute of Agricultural Technology); IFONA (Argentinean Forest Institute).

ABSTRACT A culture-independent molecular phylogenetic survey was carried out for the bacterial community in Obsidian Pool (OP), a Yellowstone National Park hot spring previously shown to contain remarkable archaeal diversity (S. M. Barns, R. E. Fundyga, M. W. Jeffries, and N. R. Page, Proc. Natl. Acad. Sci. USA 91:1609–1613, 1994). Small-subunit rRNA genes (rDNA) were amplified directly from OP sediment DNA by PCR with universally conserved orBacteria-specific rDNA primers and cloned. Unique rDNA types among >300 clones were identified by restriction fragment length polymorphism, and 122 representative rDNA sequences were determined. These were found to represent 54 distinct bacterial sequence types or clusters (≥98% identity) of sequences. A majority (70%) of the sequence types were affiliated with 14 previously recognized bacterial divisions (main phyla; kingdoms); 30% were unaffiliated with recognized bacterial divisions. The unaffiliated sequence types (represented by 38 sequences) nominally comprise 12 novel, division level lineages termed candidate divisions. Several OP sequences were nearly identical to those of cultivated chemolithotrophic thermophiles, including the hydrogen-oxidizing Calderobacterium and the sulfate reducers Thermodesulfovibrio andThermodesulfobacterium, or belonged to monophyletic assemblages recognized for a particular type of metabolism, such as the hydrogen-oxidizing Aquificales and the sulfate-reducing δ-Proteobacteria. The occurrence of such organisms is consistent with the chemical composition of OP (high in reduced iron and sulfur) and suggests a lithotrophic base for primary productivity in this hot spring, through hydrogen oxidation and sulfate reduction. Unexpectedly, no archaeal sequences were encountered in OP clone libraries made with universal primers. Hybridization analysis of amplified OP DNA with domain-specific probes confirmed that the analyzed community rDNA from OP sediment was predominantly bacterial. These results expand substantially our knowledge of the extent of bacterial diversity and call into question the commonly held notion that Archaea dominate hydrothermal environments. Finally, the currently known extent of division level bacterial phylogenetic diversity is collated and summarized.

In recent years, STEAM (Science, Technology, Engineering, Art, and Mathematics) has received wide attention. STEAM complements early childhood learning needs in honing 2nd century skills. This study aims to introduce a loose section in early childhood learning to pre-service teachers and then to explore their perceptions of how to use loose parts in supporting STEAM. The study design uses qualitative phenomenological methods. FGDs (Focus Group Discussions) are used as data collection instruments. The findings point to two main themes that emerged from the discussion: a loose section that supports freedom of creation and problem solving. Freedom clearly supports science, mathematics and arts education while problem solving significantly supports engineering and technology education. Keywords: Early Childhood Educators, Loose-part, STEAM References: Allen, A. (2016). Don’t Fear STEM: You Already Teach It! Exchange, (231), 56–59. Ansberry, B. K., & Morgan, E. (2019). Seven Myths of STEM. 56(6), 64–67. Bagiati, A., & Evangelou, D. (2015). Engineering curriculum in the preschool classroom: the teacher’s experience. European Early Childhood Education Research Journal, 23(1), 112–128. https://doi.org/10.1080/1350293X.2014.991099 Becker, K., & Park, K. (2011). Effects of integrative approaches among science , technology , engineering , and mathematics ( STEM ) subjects on students ’ learning : A preliminary meta-analysis. 12(5), 23–38. Berk, L. E. (2009). Child Development (8th ed.). Boston: Pearson Education. Can, B., Yildiz-Demirtas, V., & Altun, E. (2017). The Effect of Project-based Science Education Programme on Scientific Process Skills and Conception of Kindergargen Students. 16(3), 395–413. Casey, T., Robertson, J., Abel, J., Cairns, M., Caldwell, L., Campbell, K., … Robertson, T. (2016). Loose Parts Play. Edinburgh. Cheung, R. H. P. (2017). Teacher-directed versus child-centred : the challenge of promoting creativity in Chinese preschool classrooms. Pedagogy, Culture & Society, 1366(January), 1–14. https://doi.org/10.1080/14681366.2016.1217253 Clements, D. H., & Sarama, J. (2016). Math, Science, and Technology in the Early Grades. The Future of Children, 26(2), 75–94. Cloward Drown, K. (2014). Dramatic lay affordances of natural and manufactured outdoor settings for preschoolaged children. Dejarnette, N. K. (2018). Early Childhood Steam: Reflections From a Year of Steam Initiatives Implemented in a High-Needs Primary School. Education, 139(2), 96–112. DiGironimo, N. (2011). What is technology? Investigating student conceptions about the nature of technology. International Journal of Science Education, 33(10), 1337–1352. https://doi.org/10.1080/09500693.2010.495400 Dugger, W. E., & Naik, N. (2001). Clarifying Misconceptions between Technology Education and Educational Technology. The Technology Teacher, 61(1), 31–35. Eeuwijk, P. Van, & Zuzana, A. (2017). How to Conduct a Focus Group Discussion ( FGD ) Methodological Manual. Flannigan, C., & Dietze, B. (2018). Children, Outdoor Play, and Loose Parts. Journal of Childhood Studies, 42(4), 53–60. https://doi.org/10.18357/jcs.v42i4.18103 Fleer, M. (1998). The Preparation of Australian Teachers in Technology Education : Developing The Preparation of Australian Teachers in Technology Education : Developing Professionals Not Technicians. Asia-Pacific Journal of Teacher Education & Development, 1(2), 25–31. Freitas, H., Oliveira, M., Jenkins, M., & Popjoy, O. (1998). The focus group, a qualitative research method: Reviewing the theory, and providing guidelines to its planning. In ISRC, Merrick School of Business, University of Baltimore (MD, EUA)(Vol. 1). Gomes, J., & Fleer, M. (2019). The Development of a Scientific Motive : How Preschool Science and Home Play Reciprocally Contribute to Science Learning. Research in Science Education, 49(2), 613–634. https://doi.org/10.1007/s11165-017-9631-5 Goris, T., & Dyrenfurth, M. (n.d.). Students ’ Misconceptions in Science , Technology , and Engineering . Gull, C., Bogunovich, J., Goldstein, S. L., & Rosengarten, T. (2019). Definitions of Loose Parts in Early Childhood Outdoor Classrooms: A Scoping Review. The International Journal of Early Childhood Environmental Education, 6(3), 37. Hui, A. N. N., He, M. W. J., & Ye, S. S. (2015). Arts education and creativity enhancement in young children in Hong Kong. Educational Psychology, 35(3), 315–327. https://doi.org/10.1080/01443410.2013.875518 Jarvis, T., & Rennie, L. J. (1996). Perceptions about Technology Held by Primary Teachers in England. Research in Science & Technological Education, 14(1), 43–54. https://doi.org/10.1080/0263514960140104 Jeffers, O. (2004). How to Catch a Star. New York: Philomel Books. Kiewra, C., & Veselack, E. (2016). Playing with nature: Supporting preschoolers’ creativity in natural outdoor classrooms. International Journal of Early Childhood Environmental Education, 4(1), 70–95. Kuh, L., Ponte, I., & Chau, C. (2013). The impact of a natural playscape installation on young children’s play behaviors. Children, Youth and Environments, 23(2), 49–77. Lachapelle, C. P., Cunningham, C. M., & Oh, Y. (2019). What is technology? Development and evaluation of a simple instrument for measuring children’s conceptions of technology. International Journal of Science Education, 41(2), 188–209. https://doi.org/10.1080/09500693.2018.1545101 Liamputtong. (2010). Focus Group Methodology : Introduction and History. In Focus Group MethodoloGy (pp. 1–14). Liao, C. (2016). From Interdisciplinary to Transdisciplinary: An Arts-Integrated Approach to STEAM Education. 69(6), 44–49. https://doi.org/10.1080/00043125.2016.1224873 Lindeman, K. W., & Anderson, E. M. (2015). Using Blocks to Develop 21st Century Skills. Young Children, 70(1), 36–43. Maxwell, L., Mitchell, M., and Evans, G. (2008). Effects of play equipment and loose parts on preschool children’s outdoor play behavior: An observational study and design intervention. Children, Youth and Environments, 18(2), 36–63. McClure, E., Guernsey, L., Clements, D., Bales, S., Nichols, J., Kendall-Taylor, N., & Levine, M. (2017). How to Integrate STEM Into Early Childhood Education. Science and Children, 055(02), 8–11. https://doi.org/10.2505/4/sc17_055_02_8 McClure, M., Tarr, P., Thompson, C. M., & Eckhoff, A. (2017). Defining quality in visual art education for young children: Building on the position statement of the early childhood art educators. Arts Education Policy Review, 118(3), 154–163. https://doi.org/10.1080/10632913.2016.1245167 Mishra, L. (2016). Focus Group Discussion in Qualitative Research. TechnoLearn: An International Journal of Educational Technology, 6(1), 1. https://doi.org/10.5958/2249-5223.2016.00001.2 Monhardt, L., & Monhardt, R. (2006). Creating a context for the learning of science process skills through picture books. Early Childhood Education Journal, 34(1), 67–71. https://doi.org/10.1007/s10643-006-0108-9 Monsalvatge, L., Long, K., & DiBello, L. (2013). Turning our world of learning inside out! Dimensions of Early Childhood, 41(3), 23–30. Moomaw, S. (2012). STEM begins in the early years. School Science & Mathematics, 112(2), 57–58. Moomaw, S. (2016). Move Back the Clock, Educators: STEM Begins at Birth. School Science & Mathematics, 116(5), 237–238. Moomaw, S., & Davis, J. A. (2010). STEM Comes to Preschool. Young Cihildren, 12–18(September), 12–18. Munawar, M., Roshayanti, F., & Sugiyanti. (2019). Implementation of STEAM (Science, Technology, Engineering, Art, Mathematics)-Based Early Childhood Education Learning in Semarang City. Jurnal CERIA, 2(5), 276–285. National Research Council. (1996). National Science Education Standards. Washington, DC: National Academy of Sciences. Nicholson, S. (1972). The Theory of Loose Parts: An important principle for design methodology. Studies in Design Education Craft & Technology, 4(2), 5–12. O.Nyumba, T., Wilson, K., Derrick, C. J., & Mukherjee, N. (2018). The use of focus group discussion methodology: Insights from two decades of application in conservation. Methods in Ecology and Evolution, 9(1), 20–32. https://doi.org/10.1111/2041-210X.12860 Padilla-Diaz, M. (2015). Phenomenology in Educational Qualitative Research : Philosophy as Science or Philosophical Science ? International Journal of Educational Excellence, 1(2), 101–110. Padilla, M. J. (1990). The Science Process Skills. Research Matters - to the Science Teacher, 1(March), 1–3. Park, D. Y., Park, M. H., & Bates, A. B. (2018). Exploring Young Children’s Understanding About the Concept of Volume Through Engineering Design in a STEM Activity: A Case Study. International Journal of Science and Mathematics Education, 16(2), 275–294. https://doi.org/10.1007/s10763-016-9776-0 Rahardjo, M. M. (2019). Implementasi Pendekatan Saintifik Sebagai Pembentuk Keterampilan Proses Sains Anak Usia Dini. Scholaria: Jurnal Pendidikan Dan Kebudayaan, 9(2), 148–159. https://doi.org/10.24246/j.js.2019.v9.i2.p148-159 Robison, T. (2016). Male Elementary General Music Teachers : A Phenomenological Study. Journal of Music Teacher Education, 26(2), 77–89. https://doi.org/10.1177/1057083715622019 Rocha Fernandes, G. W., Rodrigues, A. M., & Ferreira, C. A. (2018). Conceptions of the Nature of Science and Technology: a Study with Children and Youths in a Non-Formal Science and Technology Education Setting. Research in Science Education, 48(5), 1071–1106. https://doi.org/10.1007/s11165-016-9599-6 Sawyer, R. K. (2006). Educating for innovation. 1(2006), 41–48. https://doi.org/10.1016/j.tsc.2005.08.001 Sharapan, H. (2012). ERIC - From STEM to STEAM: How Early Childhood Educators Can Apply Fred Rogers’ Approach, Young Children, 2012-Jan. Young Children, 67(1), 36–40. Siantayani, Y. (2018). STEAM: Science-Technology-Engineering-Art-Mathematics. Semarang: SINAU Teachers Development Center. Sikder, S., & Fleer, M. (2015). Small Science : Infants and Toddlers Experiencing Science in Everyday Family Life. Research in Science Education, 45(3), 445–464. https://doi.org/10.1007/s11165-014-9431-0 Smith-gilman, S. (2018). The Arts, Loose Parts and Conversations. Journal of the Canadian Association for Curriculum Studies, 16(1), 90–103. Sohn, B. K., Thomas, S. P., Greenberg, K. H., & Pollio, H. R. (2017). Hearing the Voices of Students and Teachers : A Phenomenological Approach to Educational Research. 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Oxide semiconductors have opened a new era for large-area, flexible and transparent applications. Despite the progresses, a bottleneck issue of oxide thin-film transistors (TFTs) is the instability either under bias stress or when used as a current source. Furthermore, the carrier mobility and current driving capability need to be improved for high-spec displays. It is still hugely challenging to overcome both issues using the conventional device structure and oxide semiconductor materials. Here, we review our recent work on novel oxide TFTs that show a few desirable properties. Rather than using an ohmic metal contact as the source electrode, a high work-function Schottky source contact enables depletion around the TFT source region, which results in intrinsic immunity to the negative bias illumination stress, no obvious short channel effect, and superb current saturation over a wide range of drain voltage1. The flat saturation current gives rise to an extremely high voltage gain reaching 23,000, which is, to the best of our knowledge, the highest gain ever achieved by a solid-state transistor to date. The threshold voltage is also found to remain stable under different drain voltages, in contrast to standard TFTs, which may be useful in larger-area displays where the drain voltages of the drive TFTs can differ2. Furthermore, the depletion provided by the Schottky source electrode allows utilizing semi-metal ITO to replace IGZO as the TFT channel layer, which significantly enhances the carrier mobility and current driving capability. Other related work may also be discussed in the talk including oxide Schottky diodes operating beyond 10 GHz3, oxide TFTs operating beyond 1 GHz4, significantly enhanced carrier mobility by self-assembled monolayer treatment5,6, and CMOS-like oxide-based logic circuits7,8. References Extremely high-gain source-gated transistors, J Zhang, J Wilson, G Auton, Y Wang, M Xu, Q Xin, A Song, Proceedings of the National Academy of Sciences 116 (11), 4843-4848 (2019) Comparative Study of Short-Channel Effects Between Source-Gated Transistors and Standard Thin-Film Transistors, Zhenze Wang, Li Luo, Yiming Wang, Jiawei Zhang, and Aimin Song, IEEE Transactions on Electron Devices, 69(2), 561 - 566 (2022). Flexible indium–gallium–zinc–oxide Schottky diode operating beyond 2.45 GHz, J Zhang, Y Li, B Zhang, H Wang, Q Xin, A Song, Nature communications 6 (1), 1-7 (2015). Amorphous-InGaZnO thin-film transistors operating beyond 1 GHz achieved by optimizing the channel and gate dimensions, Y Wang, J Yang, H Wang, J Zhang, H Li, G Zhu, Y Shi, Y Li, Q Wang, Qian Xin, Zhongchao Fan, Fuhua Yang, Aimin Song, IEEE Transactions on Electron Devices 65 (4), 1377-1382 (2018). Significant Performance Improvement of Oxide Thin‐Film Transistors by a Self‐Assembled Monolayer Treatment, W Cai, J Zhang, J Wilson, J Brownless, S Park, L Majewski, A Song, Advanced Electronic Materials 6 (5), 1901421 (2020). Significant performance enhancement of very thin InGaZnO thin-film transistors by a self-assembled monolayer treatment, W Cai, J Wilson, J Zhang, J Brownless, X Zhang, LA Majewski, A Song, ACS Applied Electronic Materials 2 (1), 301-308 (2020). Complementary integrated circuits based on p-type SnO and n-type IGZO thin-film transistors, Y Li, J Yang, Y Wang, P Ma, Y Yuan, J Zhang, Z Lin, L Zhou, Q Xin, Aimin Song, IEEE Electron Device Letters 39 (2), 208-211 (2017). Thin Film Sequential Circuits: Flip-Flops and a Counter Based on p-SnO and n-InGaZnO, Y Yuan, J Yang, Y Wang, Z Hu, L Zhou, Q Xin, A Song, IEEE Electron Device Letters 42 (1), 62-65 (2020).

BackgroundMyeloid-derived suppressor cells and tumor-associated macrophages inhibit anti-cancer immune responses systemically and in the tumor microenvironment, respectively, thereby limiting the efficacy of immune checkpoint blockers.1–5 However, the plasticity of myeloid cells may enable therapeutic intervention. The inhibitory receptor LILRB2/ILT4, which is expressed primarily in myeloid cells (monocytes, macrophages, dendritic cells and neutrophils), has emerged as a key immune checkpoint mediating the tolerogenic activity of myeloid cells associated with cancer.6–8 LILRB2/ILT4 has several ligands (classical and non-classical MHC-I, ANGPTL2/5, SEMA4A and CD1) and most of these are known to contribute to immune suppression in the tumor microenvironment.9–14 Thus, LILRB2/ILT4 is a promising target to overcome protumor myeloid cell activity. IO-108 is a fully human IgG4 therapeutic candidate that binds LILRB2/ILT4 with high affinity and specificity, thereby blocking its ligand interactions.MethodsWe used computational approaches to evaluate LILRB2/ILT4 expression in solid tumors from TCGA. IO-108 was discovered from a phage-displayed human single chain variable fragment antibody library. IO-108 contains the S228P mutation in the hinge region to prevent Fab-arm exchange. The binding affinity of IO-108 was measured using biolayer interferometry. The specificity of IO-108 was confirmed by two methods: 1) ELISA using recombinant LILR family members; 2) flow cytometry using cell lines engineered to express the extracellular domain of every LILR on the cell surface. Reporter and ligand binding assays were used to demonstrate LILRB2/ILT4 blocking activity of IO-108. Functional studies using primary immune cells from healthy donors and solid tumor cancer patients were performed to characterize IO-108 activity and mechanism of action. The in vivo efficacy of IO-108 is currently being evaluated in mouse models.ResultsWe found high LILRB2/ILT4 expression associated with macrophage infiltration in many solid tumor types from TCGA. IO-108 binds to LILRB2 with high affinity and specificity and blocks LILRB2/ILT4 ligand binding and activation. IO-108 enhanced the production of multiple proinflammatory cytokines in LPS- and anti-CD3- stimulated PBMC cultures from healthy donors and potentiated DC maturation/activation in response to LPS. Moreover, IO-108 polarized primary CD14+ cells isolated from solid tumor patient PBMC and ovarian cancer-associated ascites towards a proinflammatory phenotype and attenuated their suppressive effect on autologous T-cell proliferation and production of tumoricidal cytokines.ConclusionsThe preclinical characterization of IO-108, a novel LILRB2/ILT4 antagonistic antibody, demonstrates its ability to polarize tumor-associated myeloid cells towards a proinflammatory phenotype and suggests potential therapeutic benefit in tumors unresponsive to immune checkpoint blockade.AcknowledgementsWe acknowledge the funding support from National Cancer Institute (1R01 CA248736 and 2P30 CA142543), the Welch Foundation (AU-0042-20030616) and the Cancer Prevention and Research Institute of Texas (RP150551 and RP190561).Ethics ApprovalPBMCs were isolated from buffy coats of healthy donors (Interstate Blood Bank). Hematopoietic samples from cancer patients were obtained through the services of the Simmons Cancer Center‘s Tissue Management Shared Resource with IRB approved protocol (STU 102010-051). All animal work was approved and conducted under the oversight of the UT Southwestern Institutional Animal Care and Use Committee (IACUC).ReferencesHanahan D, Coussens LM. Accessories to the crime: functions of cells recruited to the tumor microenvironment. Cancer Cell 2012;21:309–22.Galluzzi L, Chan TA, Kroemer G, et al. The hallmarks of successful anticancer immunotherapy. Sci Transl Med 2018;10, DOI: 10.1126/scitranslmed.aat7807.3. Talmadge JE, Gabrilovich DI. History of myeloid-derived suppressor cells. Nat Rev Cancer 2013;13:739–52.Mantovani A, Marchesi F, Malesci A, et al. Tumour-associated macrophages as treatment targets in oncology. Nat Rev Clin Oncol 2017;14:399–416.Cassetta L, Pollard JW. Targeting macrophages: therapeutic approaches in cancer. 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Arsenic and Protein Expression: It might help to know the mechanism of As toxicity is described Introduction One of the largest public health problems at present is the drinking of water containing levels of Inorg-As that are known to be carcinogenic. The chronic ingestion of Inorg-As can results in skin cancer, urinary bladder cancer, lungs cancer, kidneys cancer, liver cancer, and cancer of other human organs 1-6. The molecular mechanisms of the carcinogenicity and toxicity of inorganic arsenic are not well understood 7–9. Many mechanisms of arsenic toxicity and carcinogenicity have been suggested 1, 7, 10 including chromosome abnormalities 11, oxidative stress 12, 13, altered growth factors 14, cell proliferation 15, altered DNA repair 16, altered DNA methylation patterns 17, inhibition of several key enzymes 18, gene amplification 19 etc. Some of these mechanisms result in alterations in protein expression. Proteomics is a powerful tool developed to enhance the study of complex biological system 20. This technique has been extensively employed to investigate the proteome response of cells to drugs and other diseases 21, 22. A proteome analysis of the Na-As (III) response in cultured lung cells found in vitro oxidative stress-induced apoptosis 23. In one of the study, hamsters were exposed to sodium arsenite (173 mg As/L) in drinking water for 6 days and several protein spots were over expressed and several were under expressed in the livers and urinary bladders of hamsters (Fig.) 24, 25. Hamsters were exposed to sodium arsenite (173 mg As/L) in drinking water for 6 days. The control hamsters were given tap water. The spot pairs of (A) equally expressed, (B) overexpressed, and (C) under expressed proteins in the liver tissues were shown. The amount of the protein is proportional to the volume of the protein peak. Transgelin was down-regulated, and GST-pi was up-regulated in the urinary bladder tissues of hamsters. In the liver tissues ornithine aminotransferase (OAT) was up-regulated, and senescence marker protein 30 (SMP 30), and fatty acid binding protein (FABP) were down-regulated. Down-regulation of transgelin has been noted in the urinary bladders of rats having bladder outlet obstruction 26. Ras-dependent and Ras-independent mechanisms can cause the down regulation of transgelin in human breast and colon carcinoma cell lines and patient-derived tumor samples 27. The loss of transgelin expression has been found in prostate cancer cells 28 and in human colonic neoplasms 29. It has been suggested that the loss of transgelin expression may be an important early event in tumor progression and a diagnostic marker for cancer development 26-29. Figure. Three-dimentional simulation of over-and under expressed protein spots in the livers of hamsters using Decyder software. Over-expression of GST-pi has been found in colon cancer tissues 30. Strong expression of GST-pi also has been found in gastric cancer 31, malignant melanoma 32, lung cancer 33, breast cancer 34 and a range of other human tumors 35. GST-pi has been up-regulated in transitional cell carcinoma of human urinary bladder 36. OAT has a role in regulating mitotic cell division and it is required for proper spindle assembly in human cancer cell 37. Ornithine amino transferase knockdown in human cervical carcinoma and osteosarcoma cells by RNA interference blocks cell division and causes cell death 37. It has been suggested that ornithine amino transferase has a role in regulating mitotic cell division and it is required for proper spindle assembly in human cancer cells 37. SMP 30 expressed mostly in the liver. By stimulating membrane calcium-pump activity it protects cells against various injuries 38. High levels of saturated, branched chain fatty acids are deleterious to cells and resulting in lipid accumulation and cytotoxicity. FABP expression has protected the cells against branched chain saturated fatty acid 39. Proteomics would be a powerful tool to know the unknown cellular mechanisms of arsenic toxicity in humans. References. NRC (National Research Council). (2001). Arsenic in Drinking Water. Update to the 1999 Arsenic in Drinking Water Report. National Academy Press, Washington, DC. Chen, C. J., Chen, C. , Wu, M. M., Kuo, T. L. (1992). Cancer potential in liver, lung, bladder, and kidney due to ingested inorganic arsenic in drinking water. Br. J. Cancer 66, 888-892. Hopenhayn-Rich, C., M.L. Biggs, A. Fuchs, et al. 1996. Bladder cancer mortality with arsenic in drinking water in Argentina. Epidemiology 7: 117–124. International Agency for Research on Cancer. (1987). In IARC Monograph on the Evaluation of Carcinogenicity Risk to Humans. 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This study aim to attempt mapping out the Land Use or Land Cover (LULC) status of Regional Project Coordination Committee (RPCC) between 2009-2019 with a view of detecting the land consumption rate and the changes that has taken place using RS and GIS techniques; serving as a precursor to the further study on urban induced variations or change in weather pattern of the cityn Rangpur City Corporation(RCC) is the main administrative functional area for both of Rangpur City and Rangpur division and experiencing a rapid changes in the field of urban sprawl, cultural and physical landscape,city growth. These agents of Land use or Land cover (LULC) varieties are responsible for multi-dimensional problems such as traffic congestion, waterlogging, and solid waste disposal, loss of agricultural land. In this regard, this study fulfills LULC changes by using Geographical Information Systems (GIS) and Remote Sensing (RS) as well as field survey was conducted for the measurement of change detection. The sources of data were Landsat 7 ETM and landsat 8 OLI/TIRS of both C1 level 1. Then after correcting the data, geometrically and radiometrically change detection and combined classification (supervised & unsupervised) were used. The study finds LULC changes built-up area, water source, agricultural land, bare soil in a change of percentage is 17.23, 2.58, -9.94, -10.19 respectively between 2009 and 2019. Among these changes, bare soil is changed to a great extent, which indicates the expansion of urban areas is utilizing the land to a proper extent. Keywords: Urban expansion; land use; land cover; remote sensing; geographic information system (GIS); Rangpur City Corporation(RCC). References Al Rifat, S. A., & Liu, W. (2019). Quantifying spatiotemporal patterns and major explanatory factors of urban expansion in miami metropolitan area during 1992-2016. Remote Sensing, 11(21) doi:10.3390/rs11212493 Arimoro AO, Fagbeja MA, Eedy W. (2002). The Need and Use of Geographic Information Systems for Environmental Impact Assessment in Africa: With Example from Ten Years Experience in Nigeria. AJEAM/RAGEE, 4(2), 16-27. Belal, A.A. and Moghanm, F.S. (2011).Detecting Urban Growth Using Remote Sensing and GIS Techniques in Al Gharbiya Governorate, Egypt.The Egyptian Journal of Remote Sensing and Space Science, 14, 73-79. http://dx.doi.org/10.1016/j.ejrs.2011.09.001 Dewan, A.M. and Yamaguchi, Y. (2009). Using Remote Sensing and GIS to Detect and Monitor and Use and Land Cover Change in Dhaka Metropolitan of Bangladesh during 1960-2005. Environmental Monitor Assessment, 150, 237- 249. Retrieved from http://dx.doi.org/10.1007/s10661-008-0226-5 Djimadoumngar, K.-N., & Adegoke, J. (2018). Satellite-Based Assessment of Land Use and Land Cover (LULC) Changes around Lake Fitri, Republic of Chad. Journal of Sustainable Development, 11(5), 71. doi:10.5539/jsd.v11n5p71 Edwards, B., Frasch, T., & Jeyacheya, J. (2019). Evaluating the effectiveness of land-use zoning for the protection of built heritage in the bagan archaeological zone, Myanmar—A satellite remote-sensing approach. Land use Policy, 88 doi:10.1016/j.landusepol.2019.104174 Fallati, L., Savini, A., Sterlacchini, S., & Galli, P. (2017). Land use and land cover (LULC) of the Republic of the Maldives: first national map and LULC change analysis using remote-sensing data. Environmental Monitoring and Assessment, 189(8). doi:10.1007/s10661-017-6120-2 Fučík, P., Novák, P., & Žížala, D. (2014). A combined statistical approach for evaluation of the effects of land use, agricultural and urban activities on stream water chemistry in small tile-drained catchments of south bohemia, czech republic. Environmental Earth Sciences, 72(6), 2195-2216. doi:10.1007/s12665-014-3131-y Elbeih, S. F., & El-Zeiny, A. M. (2018). Qualitative assessment of groundwater quality based on land use spectral retrieved indices: Case study sohag governorate, egypt. Remote Sensing Applications: Society and Environment, 10, 82-92. doi:10.1016/j.rsase.2018.03.001 Fasal, S. (2000). Urban expansion and loss of agricultural land – A GIS based study of Saharanpur City, India. Environment and Urbanization, 12(2), 133 – 149 He, S., Wang, X., Dong, J., Wei, B., Duan, H., Jiao, J., & Xie, Y. (2019). Three-dimensional urban expansion analysis of valley-type cities: A case study of chengguan district, lanzhou, china. Sustainability (Switzerland), 11(20) doi:10.3390/su11205663 Heimlich, R.E and W.D. Anderson. (2001). 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Journal of Poverty, Investment and Development, 8. Mondal, I., Srivastava, V. K., Roy, P. S., & Talukdar, G. (2014). Using logit model to identify the drivers of landuse landcover change in the lower gangetic basin, india. Paper presented at the International Archives of the Photogrammetry, Remote Sensing and Spatial Information Sciences - ISPRS Archives, , XL-8(1) 853-859. doi:10.5194/isprsarchives-XL-8-853-2014 Navale, V. B., & Mhaske, S. Y. (2019). Land use/land cover changes in sangamner city by using remote sensing and GIS. International Journal of Recent Technology and Engineering, 8(2), 4614-4621. doi:10.35940/ijrte.B3386.078219 Nicolson, L.D. (1987). The Greening of the cities; Routledge and Kegan Paul, London Nong, D., Fox, J., Miura, T., & Saksena, S. (2015). Built-up Area Change Analysis in Hanoi Using Support Vector Machine Classification of Landsat Multi-Temporal Image Stacks and Population Data. 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(2008). Impact of land use and land cover change on the water balance of a large agricultural watershed: Historical effects and future directions. Water Resources Research, 44(7). doi:10.1029/2007wr006644 Copyright (c) 2019 Geosfera Indonesia Journal and Department of Geography Education, University of Jember This work is licensed under a Creative Commons Attribution-Share A like 4.0 International License

BackgroundSystemic lupus erythematosus (SLE) is an inflammatory autoimmune disease, involving the development of autoreactive cells and autoantibodies. Natural Killer (NK) cells are innate immune cells that mediate the interaction between the innate and adaptive immune system, however their role in SLE is incompletely understood. SLE NK cells are decreased in peripheral blood, exhibit reduced cytotoxicity, and impaired cytokine production (1, 2). Furthermore, SLE NK cells present phenotypic alterations: increased expression of CD38 and altered upregulation of SLAMF7 after activation (3). To date, few studies evaluated the molecular mechanisms underlying NK cell dysfunction in SLE.ObjectivesWe examined immunometabolic alterations of SLE NK cells. First, we characterized the cellular metabolism of SLE NK cells by assessing glycolysis and oxidative phosphorylation (OXPHOS) at basal level. Then, we evaluated how cellular metabolism can be manipulated to enhance NK cell function. In this perspective, we examined how the ligation of CD38 with daratumumab (DARA) and SLAMF7 with elotuzumab (ELO) modulate glycolysis and OXPHOS.MethodsNK cells of cryopreserved PBMC from SLE patients were isolated. Glycolysis and OXPHOS were studied using XFe96 Seahorse. Expression of metabolic receptors (CD71, GLUT-1, CD98), mitochondrial function (mitochondrial membrane potential, mass) and calcium influx were investigated by FACS. Mitochondrial structure was evaluated by electron and confocal microscopy.ResultsFirst, we examined the cellular metabolism of SLE NK cells compared to healthy cells. We observed that OXPHOS is significantly increased in SLE NK cells (Figure 1A), whereas glycolysis was normal (Figure 1B). Furthermore, the mitochondrial mass and membrane potential (by FACS (Figure 1C) and confocal microscopy) were increased in SLE. Electron microscope imaging showed profound alterations in SLE NK cell mitochondrial ultrastructure (Figure 1D). No significant differences in the expression of key metabolite transporters involved in mitochondrial fueling (CD71, GLUT-1, CD98) was observed in SLE NK cells compared to healthy controls.Second, we examined how ligation of DARA and ELO influences the metabolism of healthy NK cells. Our data showed that ELO primarily enhances NK cell OXPHOS (Figure 1E), whereas DARA mainly increases glycolysis. Consistently, ELO also increases mitochondrial membrane potential and expression of metabolite transporters CD71, GLUT-1 and CD98. Next, we examined the effect of DARA and ELO on SLE NK cells. While stimulation with DARA adequately increases glycolysis in SLE NK cells, engagement with ELO fails to properly increase OXPHOS (Figure 1F), expression of cell surface transporters, mitochondrial membrane potential and mass.ConclusionOur data suggest that SLE NK cells exhibit alterations in cellular metabolism, primarily involving mitochondrial respiration. In contrast, glucose metabolism is similar to that of healthy NK cells. Additionally, ELO and DARA mediate the activation of healthy NK cells through the engagement of different metabolic pathways: OXPHOS and glycolysis, respectively. Therefore, priming SLE NK cells with ELO is unable to adequately engage their dysfunctional mitochondrial respiration. These findings provide important insights on the alteration present in SLE NK cells and contribute to a better understanding of the pathogenesis of the disease.References[1]Spada R, Rojas JM, Barber DF. Recent findings on the role of natural killer cells in the pathogenesis of systemic lupus erythematosus. J Leukoc Biol. 2015;98(4):479-87.[2]Park Y-W, Kee S-J, Cho Y-N, Lee E-H, Lee H-Y, Kim E-M, et al. Impaired differentiation and cytotoxicity of natural killer cells in systemic lupus erythematosus. Arthritis & Rheumatism. 2009;60(6):1753-63.[3]Humbel M, Bellanger F, Fluder N, Horisberger A, Suffiotti M, Fenwick C, et al. Restoration of NK Cell Cytotoxic Function With Elotuzumab and Daratumumab Promotes Elimination of Circulating Plasma Cells in Patients With SLE. Front Immunol. 2021;12:645478.AcknowledgementsThis study received funding from the Swiss National Science Foundation (Ambizione PZ00P3_173950 to DC).Disclosure of InterestsNone declared

Eating habits develop during the first years of a child's life, children learn what, when, and how much to eat through direct experience with food and by observing the eating habits of others. The aim of this study is to get a clear picture of the Eating program Healthy, starting from the planning, implementation, supervision, and evaluation as a case study of nutrition education; to get information about the advantages, disadvantages and effects of implementing a healthy eating program for children. This research was conducted through a case study with qualitative data analysed using Miles and Huberman techniques. Sample of children in Ananda Islāmic School Kindergarten. The results showed the Healthy Eating program could be implemented well, the diet was quite varied and could be considered a healthy and nutritious food. The visible impact is the emotion of pleasure experienced by children, children become fond of eating vegetables, and make children disciplined and responsible. Inadequate results were found due to the limitations of an adequate kitchen for cooking healthy food, such as cooking activities still carried out by the cook himself at the Foundation's house which is located not far from the school place; use of melamine and plastic cutlery for food; the spoon and fork used already uses aluminium material but still does not match its size; does not involve nutritionists. Keywords: Early Childhood, Eating Healthy Program References: Bandura, A. (1977). Social learning theory. Englewood Cliffs: Prentice-Hall. Bandura, Albert. (2004). Health promotion by social cognitive means. Health Education and Behavior, 31(2), 143–164. https://doi.org/10.1177/1090198104263660 Battjes-Fries, M. C. E., Haveman-Nies, A., Renes, R. J., Meester, H. J., & Van’T Veer, P. (2015). Effect of the Dutch school-based education programme “Taste Lessons” on behavioural determinants of taste acceptance and healthy eating: A quasi-experimental study. 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Hamsters were exposed to sodium arsenite (173 mg As/L) in drinking water for 6 days. Equal amounts of proteins from urinary bladder or liver extracts of control and arsenic-treated hamsters were labeled with Cy3 and Cy5 dyes, respectively. After differential in gel electrophoresis and analysis by the DeCyder software, several protein spots were found to be down-regulated and several were up regulated. Our experiments indicated that in the bladder tissues of hamsters exposed to arsenite, transgelin was down-regulated and GST-pi was up-regulated. The loss of transgelin expression has been reported to be an important early event in tumor progression and a diagnostic marker for cancer development [29-32]. Down-regulation of transgelin expression may be associated with the carcinogenicity of inorganic arsenic in the urinary bladder. In the liver of arsenite-treated hamsters, ornithine aminotransferase was up-regulated, and senescence marker protein 30 and fatty acid binding protein were down-regulated. The volume ratio changes of these proteins in the bladder and liver of hamsters exposed to arsenite were significantly different than that of control hamsters. Introduction Chronic exposure to inorganic arsenic can cause cancer of the skin, lungs, urinary bladder, kidneys, and liver [1-6]. The molecular mechanisms of the carcinogenicity and toxicity of inorganic arsenic are not well understood [7-9). Humans chronically exposed to inorganic arsenic excrete MMA(V), DMA(V) and the more toxic +3 oxidation state arsenic biotransformants MMA(III) and DMA (III) in their urine [10, 11], which are carcinogen [12]· After injection of mice with sodium arsenate, the highest concentrations of the very toxic MMA(III) and DMA(III) were in the kidneys and urinary bladder tissue, respectively, as shown by experiments of Chowdhury et al [13]. Many mechanisms of arsenic toxicity and carcinogenicity have been suggested [1, 7, 14] including chromosome abnormalities [15], oxidative stress [16, 17], altered growth factors [18], cell proliferation [19], altered DNA repair [20], altered DNA methylation patterns [21], inhibition of several key enzymes [22], gene amplification [23] etc. Some of these mechanisms result in alterations in protein expression. Methods for analyzing multiple proteins have advanced greatly in the last several years. In particularly, mass spectrometry (MS) and tandem MS (MS/MS) are used to analyze peptides following protein isolation using two-dimensional (2-D) gel electrophoresis and proteolytic digestion [24]. In the present study, Differential In Gel Electrophoresis (DIGE) coupled with Mass Spectrometry (MS) has been used to study some of the proteomic changes in the urinary bladder and liver of hamsters exposed to sodium arsenite in their drinking water. Our results indicated that transgelin was down-regulated and GST-pi was up-regulated in the bladder tissues. In the liver tissues ornithine aminotransferase was up-regulated, and senescence marker protein 30, and fatty acid binding protein were down-regulated. Materials and Methods Chemicals Tris, Urea, IPG strips, IPG buffer, CHAPS, Dry Strip Cover Fluid, Bind Silane, lodoacetamide, Cy3 and Cy5 were from GE Healthcare (formally known as Amersham Biosciences, Uppsala, Sweden). Thiourea, glycerol, SDS, DTT, and APS were from Sigma-Aldrich (St. Louis, MO, USA). Glycine was from USB (Cleveland, OH, USA). Acrylamide Bis 40% was from Bio-Rad (Hercules, CA, USA). All other chemicals and biochemicals used were of analytical grade. All solutions were made with Milli-Q water. Animals Male hamsters (Golden Syrian), 4 weeks of age, were purchased from Harlan Sprague Dawley, USA. Upon arrival, hamsters were acclimated in the University of Arizona animal care facility for at least 1 week and maintained in an environmentally controlled animal facility operating on a 12-h dark/12-h light cycle and at 22-24°C. They were provided with Teklad (Indianapolis, IN) 4% Mouse/Rat Diet # 7001 and water, ad libitum, throughout the acclimation and experimentation periods. Sample preparation and labelling Hamsters were exposed to sodium arsenite (173 mg) in drinking water for 6 days and the control hamsters were given tap water. On the 6th day hamsters were decapitated rapidly by guillotine. Urinary bladder tissues and liver were removed, blotted on tissue papers (Kimtech Science, Precision Wipes), and weighed. Hamster urinary bladder or liver tissues were homogenized in lysis buffer (30mMTris, 2M thiourea, 7M urea, and 4% w/w CHAPS adjusted to pH 8.5 with dilute HCI), at 4°C using a glass homogenizer and a Teflon coated steel pestle; transferred to a 5 ml acid-washed polypropylene tube, placed on ice and sonicated 3 times for 15 seconds. The sonicate was centrifuged at 12,000 rpm for 10 minutes at 4°C. Small aliquots of the supernatants were stored at -80°C until use (generally within one week). Protein concentration was determined by the method of Bradford [25] using bovine serum albumin as a standard. Fifty micrograms of lysate protein was labeled with 400 pmol of Cy3 Dye (for control homogenate sample) and Cy5 Dye (for arsenic-treated urinary bladder or liver homogenate sample). The samples containing proteins and dyes were incubated for 30 min on ice in the dark. To stop the labeling reaction, 1uL of 10 mM lysine was added followed by incubation for 10 min on ice in the dark. To each of the appropriate dye-labeled protein samples, an additional 200 ug of urinary bladderor liver unlabeled protein from control hamster sample or arsenic-treated hamster sample was added to the appropriate sample. Differentially labeled samples were combined into a single Microfuge tube (total protein 500 ug); protein was mixed with an equal volume of 2x sample buffer [2M thiourea, 7M urea, pH 3-10 pharmalyte for isoelectric focusing 2% (v/v), DTT 2% (w/v), CHAPS 4% (w/v)]; and was incubated on ice in the dark for 10 min. The combined samples containing 500 ug of total protein were mixed with rehydration buffer [CHAPS 4% (w/v), 8M urea, 13mM DTT, IPG buffer (3-10) 1% (v/v) and trace amount of bromophenol blue]. The 450 ul sample containing rehydration buffer was slowly pipetted into the slot of the ImmobilinedryStripReswelling Tray and any large bubbles were removed. The IPG strip (linear pH 3-10, 24 cm) was placed (gel side down) into the slot, covered with drystrip cover fluid (Fig. 1), and the lid of the Reswelling Tray was closed. The ImmobillineDryStrip was allowed to rehydrate at room temperature for 24 hours. First dimension Isoelectric focusing (IEF) The labeled sample was loaded using the cup loading method on universal strip holder. IEF was then carried out on EttanIPGphor II using multistep protocol (6 hr @ 500 V, 6 hr @ 1000 V, 8 hr @ 8000 V). The focused IPG strip was equilibrated in two steps (reduction and alkylation) by equilibrating the strip for 10 min first in 10 ml of 50mM Tris (pH 8.8), 6M urea, 30% (v/v) glycerol, 2% (w/v) SDS, and 0.5% (w/v) DTT, followed by another 10 min in 10 ml of 50mM Tris (pH 8.8), 6M urea, 30% (v/v) glycerol, 2% (w/v) SDS, and 4.5% (w/v) iodoacetamide to prepare it for the second dimension electrophoresis. Second dimension SDS-PAGE The equilibrated IPG strip was used for protein separation by 2D-gel electrophoresis (DIGE). The strip was sealed at the top of the acrylamide gel for the second dimension (vertical) (12.5% polyacrylamide gel, 20x25 cm x 1.5 mm) with 0.5% (w/v) agarose in SDS running buffer [25 mMTris, 192 mM Glycine, and 0.1% (w/v) SDS]. Electrophoresis was performed in an Ettan DALT six electrophoresis unit (Amersham Biosciences) at 1.5 watts per gel, until the tracking dye reached the anodic end of the gel. Image analysis and post-staining The gel then was imaged directly between glass plates on the Typhoon 9410 variable mode imager (Sunnyvale, CA, USA) using optimal excitation/emission wavelength for each DIGE fluor: Cy3 (532/580 nm) and Cy5 (633/670 nm). The DIGE images were previewed and checked with Image Quant software (GE Healthcare) where all the two separate gel images could be viewed as a single gel image. DeCyde v.5.02 was used to analyze the DIGE images as described in the Ettan DIGE User Manual (GE Healthcare). The appropriate up-/down regulated spots were filtered based on an average volume ratio of ± over 1.2 fold. After image acquisition, the gel was fixed overnight in a solution containing 40% ethanol and 10% acetic acid. The fixed gel was stained with SyproRuby (BioRad) according to the manufacturer protocol (Bio-Rad Labs., 2000 Alfred Nobel Drive, Hercules, CA 94547). Identification of proteins by MS Protein spot picking and digestion Sypro Ruby stained gels were imaged using an Investigator ProPic and HT Analyzer software, both from Genomic Solutions (Ann Arbor, MI). Protein spots of interest that matched those imaged using the DIGE Cy3/Cy5 labels were picked robotically, digested using trypsin as described previously [24] and saved for mass spectrometry identification. Liquid chromatography (LC)- MS/MS analysis LC-MS/MS analyses were carried out using a 3D quadrupole ion trap massspectrometer (ThermoFinnigan LCQ DECA XP PLUS; ThermoFinnigan, San Jose, CA) equipped with a Michrom Paradigm MS4 HPLC (MichromBiosources, Auburn, CA) and a nanospray source, or with a linear quadrupole ion trap mass spectrometer (ThermoFinnigan LTQ), also equipped with a Michrom MS4 HPLC and a nanospray source. Peptides were eluted from a 15 cm pulled tip capillary column (100 um I.D. x 360 um O.D.; 3-5 um tip opening) packed with 7 cm Vydac C18 (Vydac, Hesperia, CA) material (5 µm, 300 Å pore size), using a gradient of 0-65% solvent B (98% methanol/2% water/0.5% formic acid/0.01% triflouroacetic acid) over a 60 min period at a flow rate of 350 nL/min. The ESI positive mode spray voltage was set at 1.6 kV, and the capillary temperature was set at 200°C. Dependent data scanning was performed by the Xcalibur v 1.3 software on the LCQ DECA XP+ or v 1.4 on the LTQ [27], with a default charge of 2, an isolation width of 1.5 amu, an activation amplitude of 35%, activation time of 50 msec, and a minimal signal of 10,000 ion counts (100 ion counts on the LTQ). Global dependent data settings were as follows: reject mass width of 1.5 amu, dynamic exclusion enabled, exclusion mass width of 1.5 amu, repeat count of 1, repeat duration of a min, and exclusion duration of 5 min. Scan event series were included one full scan with mass range of 350-2000 Da, followed by 3 dependent MS/MS scans of the most intense ion. Database searching Tandem MS spectra of peptides were analyzed with Turbo SEQUEST, version 3.1 (ThermoFinnigan), a program that allows the correlation of experimental tandem MS data with theoretical spectra generated from known protein sequences. All spectra were searched against the latest version of the non redundant protein database from the National Center for Biotechnology Information (NCBI 2006; at that time, the database contained 3,783,042 entries). Statistical analysis The means and standard error were calculated. The Student's t-test was used to analyze the significance of the difference between the control and arsenite exposed hamsters. P values less than 0.05 were considered significant. The reproducibility was confirmed in separate experiments. Results Analysis of proteins expression After DIGE (Fig. 1), the gel was scanned by a Typhoon Scanner and the relative amount of protein from sample 1 (treated hamster) as compared to sample 2 (control hamster) was determined (Figs. 2, 3). A green spot indicates that the amount of protein from sodium arsenite-treated hamster sample was less than that of the control sample. A red spot indicates that the amount of protein from the sodium arsenite-treated hamster sample was greater than that of the control sample. A yellow spot indicates sodium arsenite-treated hamster and control hamster each had the same amount of that protein. Several protein spots were up-regulated (red) or down-regulated (green) in the urinary bladder samples of hamsters exposed to sodium arsenite (173 mg As/L) for 6 days as compared with the urinary bladder of controls (Fig. 2). In the case of liver, several protein spots were also over-expressed (red) or under-expressed (green) for hamsters exposed to sodium arsenite (173 mg As/L) in drinking water for 6 days (Fig. 3). The urinary bladder samples were collected from the first and second experiments in which hamsters were exposed to sodium arsenite (173 mg As/L) in drinking water for 6 days and the controls were given tap water. The urinary bladder samples from the 1st and 2nd experiments were run 5 times in DIGE gels on different days. The protein expression is shown in Figure 2 and Table 1. The liver samples from the 1st and 2nd experiments were also run 3 times in DIGE gels on different days. The proteins expression were shown in Figure 3 and Table 2. The volume ratio changed of the protein spots in the urinary bladder and liver of hamsters exposed to arsenite were significantly differences than that of the control hamsters (Table 1 and 2). Protein spots identified by LC-MS/MS Bladder The spots of interest were removed from the gel, digested, and their identities were determined by LC-MS/MS (Fig. 2 and Table 1). The spots 1, 2, & 3 from the gel were analyzed and were repeated for the confirmation of the results (experiments; 173 mg As/L). The proteins for the spots 1, 2, and 3 were identified as transgelin, transgelin, and glutathione S-transferase Pi, respectively (Fig. 2). Liver We also identified some of the proteins in the liver samples of hamsters exposed to sodium arsenite (173 mg As/L) in drinking water for 6 days (Fig. 3). The spots 4, 5, & 6 from the gels were analyzed and were repeated for the confirmation of the results. The proteins for the spots 4, 5, and 6 were identified as ornithine aminotransferase, senescence marker protein 30, and fatty acid binding protein, respectively (Fig. 3) Discussion The identification and functional assignment of proteins is helpful for understanding the molecular events involved in disease. Weexposed hamsters to sodium arsenite in drinking water. Controls were given tap water. DIGE coupled with LC-MS/MS was then used to study the proteomic change in arsenite-exposed hamsters. After electrophoresis DeCyder software indicated that several protein spots were down-regulated (green) and several were up-regulated (red). Our overall results as to changes and functions of the proteins we have studied are summarized in Table 3. Bladder In the case of the urinary bladder tissue of hamsters exposed to sodium arsenite (173 mg As/L) in drinking water for 6 days, transgelin was down-regulated and GST-pi was up-regulated. This is the first evidence that transgelin is down-regulated in the bladders of animals exposed to sodium arsenite. Transgelin, which is identical to SM22 or WS3-10, is an actin cross linking/gelling protein found in fibroblasts and smooth muscle [28, 29]. It has been suggested that the loss of transgelin expression may be an important early event in tumor progression and a diagnostic marker for cancer development [30-33]. It may function as a tumor suppressor via inhibition of ARA54 (co-regulator of androgen receptor)-enhanced AR (androgen receptor) function. Loss of transgelin and its suppressor function in prostate cancer might contribute to the progression of prostate cancer [30]. Down-regulation of transgelin occurs in the urinary bladders of rats having bladder outlet obstruction [32]. Ras-dependent and Ras-independent mechanisms can cause the down regulation of transgelin in human breast and colon carcinoma cell lines and patient-derived tumorsamples [33]. Transgelin plays a role in contractility, possibly by affecting the actin content of filaments [34]. In our experiments loss of transgelin expression may be associated or preliminary to bladder cancer due to arsenic exposure. Arsenite is a carcinogen [1]. In our experiments, LC-MS/MS analysis showed that two spots (1 and 2) represent transgelin (Fig. 2 and Table 1). In human colonic neoplasms there is a loss of transgelin expression and the appearance of transgelin isoforms (31). GST-pi protein was up-regulated in the bladders of the hamsters exposed to sodium arsenite. GSTs are a large family of multifunctional enzymes involved in the phase II detoxification of foreign compounds [35]. The most abundant GSTS are the classes alpha, mu, and pi classes [36]. They participate in protection against oxidative stress [37]. GST-omega has arsenic reductase activity [38]. Over-expression of GST-pi has been found in colon cancer tissues [39]. Strong expression of GST-pi also has been found in gastric cancer [40], malignant melanoma [41], lung cancer [42], breast cancer [43] and a range of other human tumors [44]. GST-pi has been up-regulated in transitional cell carcinoma of human urinary bladder [45]. Up-regulation of glutathione – related genes and enzyme activities has been found in cultured human cells by sub lethal concentration of inorganic arsenic [46]. There is evidence that arsenic induces DNA damage via the production of ROS (reactive oxygen species) [47]. GST-pi may be over-expressed in the urinary bladder to protect cells against arsenic-induced oxidative stress. Liver In the livers of hamsters exposed to sodium arsenite, ornithine amino transferase was over-expressed, senescence marker protein 30 was under-expressed, and fatty acid binding protein was under-expressed. Ornithine amino transferase has been found in the mitochondria of many different mammalian tissues, especially liver, kidney, and small intestine [48]. Ornithine amino transferase knockdown inhuman cervical carcinoma and osteosarcoma cells by RNA interference blocks cell division and causes cell death [49]. It has been suggested that ornithine amino transferase has a role in regulating mitotic cell division and it is required for proper spindle assembly in human cancer cells [49]. Senescence marker protein-30 (SMP30) is a unique enzyme that hydrolyzes diisopropylphosphorofluoridate. SMP30, which is expressed mostly in the liver, protects cells against various injuries by stimulating membrane calcium-pump activity [50]. SMP30 acts to protect cells from apoptosis [51]. In addition it protects the liver from toxic agents [52]. The livers of SMP30 knockout mice accumulate phosphatidylethanolamine, cardiolipin, phosphatidyl-choline, phosphatidylserine, and sphingomyelin [53]. Liver fatty acid binding protein (L-FABP) also was down- regulated. Decreased liver fatty acid-binding capacity and altered liver lipid distribution hasbeen reported in mice lacking the L-FABP gene [54]. High levels of saturated, branched-chain fatty acids are deleterious to cells and animals, resulting in lipid accumulation and cytotoxicity. The expression of fatty acid binding proteins (including L-FABP) protected cells against branched-chain saturated fatty acid toxicity [55]. Limitations: we preferred to study the pronounced spots seen in DIGE gels. Other spots were visible but not as pronounced. Because of limited funds, we did not identify these others protein spots. In conclusion, urinary bladders of hamsters exposed to sodium arsenite had a decrease in the expression of transgelin and an increase in the expression of GST-pi protein. Under-expression of transgelin has been found in various cancer systems and may be associated with arsenic carcinogenicity [30-33). Inorganic arsenic exposure has resulted in bladder cancer as has been reported in the past [1]. Over-expression of GST-pi may protect cells against oxidative stress caused by arsenite. In the liver OAT was up regulated and SMP-30 and FABP were down regulated. These proteomic results may be of help to investigators studying arsenic carcinogenicity. The Superfund Basic Research Program NIEHS Grant Number ES 04940 from the National Institute of Environmental Health Sciences supported this work. Additional support for the mass spectrometry analyses was provided by grants from NIWHS ES06694, NCI CA023074 and the BIOS Institute of the University of Arizona. Acknowledgement The Author wants to dedicate this paper to the memory of his former supervisor Dr. H. VaskenAposhian who passed away in September 6, 2019. He was an emeritus professor of the Department of Molecular and Cellular Biology at the University of Arizona. 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In the last few decades, the studies in second language acquisition have not answered the question what mechanisms a human’s brain uses to make acquisition of language(s) possible. A neurocognitive model which tries to address SLA from such a perspective was suggested by Ullman (2005; 2015), according to which, “both first and second languages are acquired and processed by well-studied brain systems that are known to subserve particular nonlanguage functions” (Ullman, 2005: 141). The brain systems in question have analogous roles in their language and nonlanguage functions. This article is meant to critically analyse the suggested DP model within the context of neurocognitive studies of L2; and evaluate its contribution to the field of SLA studies. References Aboitiz, F. (1995). Working memory networks and the origin of language areas in the human brain. Medical Hypothesis, 25, 504-506. Aboitiz, F. & Garcia, R. (1977). The anatomy of language revisited. 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The pandemic of COVID-19 has afflicted every individual and has initiated a cascade of directly or indirectly involved events in precipitating mental health issues. The human species is a wanderer and hunter-gatherer by nature, and physical social distancing and nationwide lockdown have confined an individual to physical isolation. The present review article was conceived to address psychosocial and other issues and their aetiology related to the current pandemic of COVID-19. The elderly age group has most suffered the wrath of SARS-CoV-2, and social isolation as a preventive measure may further induce mental health issues. Animal model studies have demonstrated an inappropriate interacting endogenous neurotransmitter milieu of dopamine, serotonin, glutamate, and opioids, induced by social isolation that could probably lead to observable phenomena of deviant psychosocial behavior. Conflicting and manipulated information related to COVID-19 on social media has also been recognized as a global threat. Psychological stress during the current pandemic in frontline health care workers, migrant workers, children, and adolescents is also a serious concern. Mental health issues in the current situation could also be induced by being quarantined, uncertainty in business, jobs, economy, hampered academic activities, increased screen time on social media, and domestic violence incidences. The gravity of mental health issues associated with the pandemic of COVID-19 should be identified at the earliest. Mental health organization dedicated to current and future pandemics should be established along with Government policies addressing psychological issues to prevent and treat mental health issues need to be developed. References World Health Organization (WHO) Coronavirus Disease (COVID-19) Dashboard. 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Our studies have indicated that the relative concentration of Se or Hg to As in urine and blood positively correlates with percentage of inorganic arsenic (% Inorg-As) and percentage of monomethlyarsonic acid [% MMA (V)]. We also found a negative correlation with percentage of dimethylarsinic acid [% DMA (V)] and the ratio of % DMA (V) to % MMA (V). In another study, we found that a group of proteins were significantly over expressed and conversely other groups were under-expressed in tissues in Na-As (III) treated hamsters. Introduction.Inorganic arsenic (Inorg-As) in drinking water.One of the largest public health problems at present is the drinking of water containing levels of Inorg-As that are known to be carcinogenic. At least 200 million people globally are at risk of dying because of arsenic (As) in their drinking water1-3. The chronic ingestion of Inorg-As can results in skin cancer, bladder cancer, lung cancer, and cancer of other organs1-3. The maximum contamination level (MCL) of U.S. drinking water for arsenic is 10 ug/L. The arsenic related public health problem in the U.S. is not at present anywhere near that of India4, Bangladesh4, and other countries5. Metabolism and toxicity of Inorg-As and arsenic species.Inorg-As is metabolized in the body by alternating reduction of pentavalent arsenic to trivalent form by enzymes and addition of a methyl group from S-adenosylmethionine6, 7; it is excreted mainly in urine as DMA (V)8. Inorganic arsenate [Inorg-As (V)]is biotransformed to Inorg-As (III), MMA (V), MMA (III), DMA (V), and DMA (III)6(Fig. 1). Therefore, the study of the toxicology of Inorg-As (V) involves at least these six chemical forms of arsenic. Studies reported the presence of 3+ oxidation state arsenic biotransformants [MMA (III) and DMA (III)] in human urine9and in animal tissues10. The MMA (III) and DMA (III) are more toxic than other arsenicals11, 12. In particular MMA (III) is highly toxic11, 12. In increased % MMA in urine has been recognized in arsenic toxicity13. In addition, people with a small % MMA in urine show less retention of arsenic14. Thus, the higher prevalence of toxic effects with increased % MMA in urine could be attributed to the presence of toxic MMA (III) in the tissue. Previous studies also indicated that males are more susceptible to the As related skin effects than females13, 15. A study in the U.S population reported that females excreted a lower % Inorg-As as well as % MMA, and a higher % DMA than did males16. Abbreviation: SAM, S-adenosyl-L-methionine; SAHC, S-adenosyl-L-homocysteine. Differences in susceptibility to arsenic toxicity might be manifested by differences in arsenic metabolism among people. Several factors (for examples, genetic factors, sex, duration and dosage of exposure, nutritional and dietary factors, etc.) could be influence for biotransformation of Inorg-As,6, 17 and other unknown factors may also be involved. The interaction between As, Se, and Hg.The toxicity of one metal or metalloid can be dramatically modulated by the interaction with other toxic and essential elements18. Arsenic and Hg are toxic elements, and Se is required to maintain good health19. But Se is also toxic at high levels20. Recent reports point out the increased risk of squamous cell carcinoma and non-melanoma skin cancer in those treated with 200 ug/day of selenium (Nutritional Prevention of Cancer Trial in the United States)21. However, it is well known that As and Se as well as Se and Hg act as antagonists22. It was also reported that Inorg-As (III) influenced the interaction between selenite and methyl mercury23. A possible molecular link between As, Se, and Hg has been proposed by Korbas et al. (2008)24. The identifying complexes between the interaction of As and Se, Se and Hg as well as As, Se, and Hg in blood of rabbit are shown in Table 1. Influence of Se and Hg on the metabolism of Inorg-As.The studies have reported that Se supplementation decreased the As-induced toxicity25, 26. The concentrations of urinary Se expressed as ug/L were negatively correlated with urinary % Inorg-As and positively correlated with % DMA27. The study did not address the urinary creatinine adjustment27. Other researchers suggested that Se and Hg decreased As methylation28-31(Table 2). They also suggested that the synthesis of DMA from MMA might be more susceptible to inhibition by Se (IV)29 as well as by Hg (II)30,31 compared to the production of MMA from Inorg-As (III). The inhibitory effects of Se and Hg were concentration dependent28-31. The literature suggests that reduced methylation capacity with increased % MMA (V), decreased % DMA (V), or decreased ratios of % DMA to % MMA in urine is positively associated with various lesions32. Lesions include skin cancer and bladder cancer32. The results were obtained from inorganic arsenic exposed subjects32. Our concern involves the combination of low arsenic (As) and high selenium (Se) ingestion. This can inhibit methylation of arsenic to take it to a toxic level in the tissue. Dietary sources of Se and Hg.Global selenium (Se) source are vegetables in the diet. In the United States, meat and bread are the common source. Selenium deficiency in the US is rare. The US Food and Drug Administration (FDA) has found toxic levels of Se in dietary supplements, up to 200 times greater than the amount stated on the label33. The samples contained up to 40,800 ug Se per recommended serving. For the general population, the most important pathway of exposure to mercury (Hg) is ingestion of methyl mercury in foods. Fish (including tuna, a food commonly eaten by children), other seafood, and marine mammals contain the highest concentrations. The FDA has set a maximum permissible level of 1 ppm of methyl mercury in the seafood34. The people also exposed mercury via amalgams35. Proteomic study of Inorg-As (III) injury.Proteomics is a powerful tool developed to enhance the study of complex biological system36. This technique has been extensively employed to investigate the proteome response of cells to drugs and other diseases37, 38. A proteome analysis of the Na-As (III) response in cultured lung cells found in vitro oxidative stress-induced apoptosis39. However, to our knowledge, no in vivo proteomic study of Inorg-As (III) has yet been conducted to improve our understanding of the cellular proteome response to Inorg-As (III) except our preliminary study 40. Preliminary Studies: Results and DiscussionThe existing data (Fig. 1) from our laboratory and others show the complex nature of Inorg-As metabolism. For many years, the major way to study, arsenic (As) metabolism was to measure InorgAs (V), Inorg-As (III), MMA (V), and DMA (V) in urine of people chronically exposed to As in their drinking water. Our investigations demonstrated for the first time that MMA (III) and DMA (III) are found in human urine9. Also we have identified MMA (III) and DMA (III) in the tissues of mice and hamsters exposed to sodium arsenate [Na-As (V)]10, 41. Influence of Se as well as Hg on the As methyltransferase.We have reported that Se (IV) as well as mercuric chloride (HgCl2) inhibited As (III) methyltransferase and MMA (III) methyltransferase in rabbit liver cytosol. Mercuric chloride was found to be a more potent inhibitor of MMA (III) methyltransferase than As (III) methyltransferase30. These results suggested that Se and Hg decreased arsenic methylation. The inhibitory effects of Se and Hg were concentration dependent30. Influence of Se and Hg in urine and blood on the percentage of urinary As metabolites.Our human studies indicated that the ratios of the concentrations of Se or Hg to As in urine and blood were positively correlated with % Inorg-As and % MMA (V). But it negatively correlated with % DMA (V) and the ratios of % DMA (V) to % MMA (V) in urine of both males and females (unpublished data) (Table 3). These results confirmed that the inhibitory effects of Se as well as Hg for the methylation of Inorg-As in humans were concentration dependent. We also found that the concentrations of Se and Hg were negatively correlated with % Inorg-As and % MMA (V). Conversely it correlated positively with % DMA (V) and the ratios of % DMA (V) to % MMA (V) in urine of both sexes (unpublished data). These correlations were not statistically significant when urinary concentrations of Se and Hg were adjusted for urinary creatinine (Table 3). Interactions of As, Se, Hg and its relationship with methylation of arsenic are summarized in Figure 2. Sex difference distribution of arsenic species in urine.Our results indicate that females have more methylation capacity of arsenic as compared to males. In our human studies (n= 191) in Mexico, we found that females (n= 98) had lower % MMA (p<0.001) and higher % DMA (p=0.006) when compared to males (n= 93) (Fig. 3). The means ratio of % MMA (V) to % Inorg-As and % DMA (V) to %MMA (V) were also lower (p<0.05) and higher (p<0.001), respectively in females compared to males. The protein expression profiles in the tissues of hamsters exposed to Na-As (III).In our preliminary studies40, hamsters were exposed to Na-As (III) (173 pg/ml as As) in their drinking water for 6 days and control hamsters were given only the water used to make the solutions for the experimental animals. After DIGE (Two-dimensional differential in gel electrophoresis) and analysis by the DeCyder software, several protein spots were found to be over-expressed (red spot) and several were under expressed (green spot) as compared to control (Figs. 4a-c). Three proteins (one was over-expressed and two were under-expressed) of each tissue (liver and urinary bladder) were identified by LC-MS/MS (liquid chromatography-tandem mass spectrometry).DIGE in combination with LC-MS/MS is a powerful tool that may help cancer investigators to understand the molecular mechanisms of cancer progression due to Inorg-As. Propose a new researchThese results suggested that selenium (Se) as well as mercury (Hg) may influence the methylation of Inorg-As and this influence could be dependent on the concentration of Se, Hg and/or the sex of the animal. Our study also suggested that the identification and functional assignment of the expressed proteins in the tissues of Inorg-As (III) exposed animals will be useful for understanding and helping to formulate a theory dealing with the molecular events of arsenic toxicity and carcinogenicity.Therefore, it would be very useful if we could do a research study with combination of Inorg-arsenic, Se, and Hg. The new research protocol could be the following:For metabolic processing, hamsters provide a good animal model. For carcinogenesis, mouse model is well accepted. The aims of this project are: 1) To map the differential distributions of arsenic (As) metabolites/species in relation to selenium (Se) and mercury (Hg) levels in male and female hamsters and 2) To chart the protein expression profile and identify the defense proteins in mice and hamsters after As injury. Experimental hamsters (male or female) will include four groups. The first group will be treated with Na arseniteNa-As(III), the second group with Na-As (III) and Na-selenite (Na-Se (IV)], the third group with Na As (III) and methyl mercuric chloride (MeHgCl), and the final group with Na-As (III), Na-Se (IV), and MeHgci at different levels. Urine and tissue will be collected at different time periods and measured for As species using high performance liquid chromatography/inductively coupled plasma-mass spectrometry (HPLC/ICP-MS). For proteomics, mice (male and female) and hamsters (male and female) will be exposed to Na-As (III)at different levels in tap water, and control mice and hamsters will be given only the tap water. Tissue will be harvested at different time periods. TWO dimensional differential in gel electrophoresis (2D-DIGE) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) will be employed to identify the expressed protein. In summary, we intend to extend our findings to: 1) Differential distribution of As metabolites in kidney, liver, lung, and urinary bladder of male and female hamsters exposed to Na-As (III), and combined with Na-As (III) and Na-Se (IV) and/or MeHgCl at different levels and different time periods, 2) Show the correlation of As species distribution in the tissue and urine for both male and female hamsters treated with and without Na-Se (IV) and/or MeHgCl, and 3) Show protein expression profile and identify the defense proteins in the tissues (liver, lung, and urinary bladder epithelium) in mice after arsenic injury. The significance of this study: The results of which have the following significances: (A) Since Inorg-As is a human carcinogen, understanding how its metabolism is influenced by environmental factors may help understand its toxicity and carcinogenicity, (B) The interactions between arsenic (As), selenium (Se), and mercury (Hg) are of practical significance because populations in various parts of the world are simultaneously exposed to Inorg-As & Se and/or MeHg, (C) These interactions may inhibit the biotransformation of Inorg-As (III) which could increase the amount and toxicity of Inorg-As (III) and MMA (III) in the tissues, (D) Determination of arsenic species profile in the tissues after ingestion of Inorg-As (III), Se (IV), and/or MeHg+ will help understand the tissue specific influence of Se and Hg on Inorg-As (III) metabolism, (E) Correlation of arsenic species between tissue and urine might help to understand the tissue burden of arsenic species when researchers just know the distribution of arsenic species in urine, (F) The identification of the defense proteins (over-expressed and under-expressed) in the tissues of the mouse may lead to understanding the mechanisms of inorganic arsenic injury in human. 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ABSTRAK Mengetahui pengaruh 3 komponen Intellectual Capital yang terdiri dari Value Added Capital Employed (VACA), Value Added Human Capital (VAHU), dan Structural Capital Value Added (STVA) dan mengetahui pengaruh Rate of Growth of Intellectual Capital (ROGIC)terhadap proporsi pembiayaan bagi hasil bank umum syariah di Indonesia periode 2015-2020 merupakan tujuan dari penelitian ini. Menggunakan regresi data panel serta data sekunder dengan mengumpulkan laporan tahunan bank umum syariah. Memanfaatkan Sampel 9 bank umum syariah yang terdaftar pada OJK dengan diseleksi menggunakan metode purposive sampling. Variabel Intellectual Capital diukur dengan metode Islamic Banking Value Added Intellectual Coefficient (IB-VAIC). Sedangkan untuk variabel proporsi pembiayaan bagi hasil menggunakan pengukuran profit sharing ratio. Hasil penelitian menunjukkan bahwa Value Added Capital Employed (VACA), Value Added Human Capital (VAHU), dan Structural Capital Value Added (STVA) dan Rate of Growth of Intellectual Capital (ROGIC) berpengaruh signifikan terhadap proporsi pembiayaan bagi hasil. Secara parsial, Value Added Capital Employed (VACA) berpengaruh signifikan negatif terhadap proporsi pembiayaan bagi hasil. Value Added Human Capital (VAHU) dan Structural Capital Value Added (STVA) memiliki pengaruh signifikan dengan arah positif pada proporsi pembiayaan bagi hasil. Rate of Growth of Intellectual Capital (ROGIC) tidak memiliki pengaruh pada proporsi pembiayaan bagi hasil. Kata Kunci: Intellectual Capital, VACA, VAHU, STVA, ROGIC, Proporsi Pembiayaan Bagi Hasil, Bank Umum Syariah. ABSTRACT Find out the effect of 3 components of Intellectual Capital, consisting of Value Added Capital Employed (VACA), Value Added Human Capital (VAHU), and Structural Capital Value Added (STVA), and find out the effect of the Rate of Growth of Intellectual Capital (ROGIC) on profit-loss sharing financing proportion of Islamic Banks in Indonesia from 2015-2020 is the goal of this research. Using panel data regression and secondary data by collecting the annual reports of Islamic Commercial Banks. Utilizing a sample of 9 Islamic Commercial Banks registered in OJK and selected using a purposive sampling method. The Intellectual Capital was measured by the Islamic Banking Value Added Intellectual Coefficient (IB-VAIC) method. Profit loss sharing financing proportion is measured by the profit sharing ratio. The results of this research showed that Value Added Capital Employed (VACA), Value Added Human Capital (VAHU), Structural Capital Value Added (STVA), and Rate of Growth of Intellectual Capital (ROGIC) had a significant effect on profit-sharing financing proportion. Partially, Value Added Capital Employed (VACA) has a negative significant effect on profit-loss sharing financing proportion. Value Added Human Capital (VAHU) and Structural Capital Value Added (STVA) have a positive significant effect on profit-loss sharing financing proportion. The rate of Growth of Intellectual Capital (ROGIC) does not affect the profit-sharing financing proportion. Keywords: Intellectual Capital, VACA, VAHU, STVA, ROGIC, Profit Loss Sharing Financing Proportion, Islamic Banks. DAFTAR PUSTAKA Aryee, S., Walumbwa, F. O., Seidu, E. Y. M., & Otaye, L. E. (2016). Developing and leveraging human capital resource to promote service quality: Testing a theory of performance. 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Starch quality is one of the most important agronomic traits in rice (Oryza sativa L). In this study, we identified single nucleotide polymorphisms (SNPs) in the Waxy and Alk genes of eight rice varieties and their associations with starch physicochemical properties.vi.e.vamylose content (AC) and gelatinization temperature (GT). Seven Sri Lankan rice varieties, Pachchaperumal, Herathbanda, At 354, Bg 352, Balasuriya, H 6 and Bw 295-5 were detected as high amylose varieties while Nipponbare exhibited low amylose content. In silico analysis of the Waxy gene revealed that all tested Sri Lankan varieties possessed ‘G’ (Wxa allele) instead of ‘T’ in the first intron which could explain varieties with high and intermediate amylose content. All Sri Lankan varieties had ‘A’ instead of ‘C’ in exon 6 of the Waxy gene and this fact was tally with the varieties showing high amylose content. Therefore, possessing the Wxa allele in the first intron and ‘A’ in exon 6 could be used as a molecular marker for the selection of high amylose varieties as validated using several Sri Lankan varieties. All Sri Lankan varieties except, Bw 295-5 exhibited the intermediate type of GT which could not be explained using the so far reported allelic differences in the Alk gene. However, Bw 295-5 which is a low GT variety had two nucleotide polymorphisms in the last exon of the Alk gene, i.e. ‘G’ and ‘TT’ that represent low GT class. Therefore, it can be concluded that sequence variations of Waxy and Alk genes reported in this study are useful in breeding local rice varieties with preferential amylose content and GT class.Key word Alk gene, amylose content, single nucleotide polymorphism, Waxy gene.INTRODUCTIONRice (Oryza sativa L.) is one of the leading food crops of the world. More than half of the world’s population relies on rice as the major daily source of calories and protein (Sartaj and Suraweera, 2005). After grain yield, quality is the most important aspect of rice breeding. Grain size and shape largely determine the market acceptability of rice, while cooking quality is influenced by the properties of starch. In rice grains starch is the major component that primarily controls rice quality. Starch consists of two forms of glucose polymers, relatively unbranched amylose and a highly branched amylopectin. Starch-synthesizing genes may contribute to variation in starch physicochemical properties because they affect the amount and structure of amylose and amylopectin in rice grain (Kharabian-Masouleh et al., 2012). Amylose content (AC), gelatinization temperature (GT) and gel consistency (GC) is the three most important determinants of eating and cooking quality. Amylose content is the ratio of amylose amount present in endosperm to total starch content. Rice varieties are grouped based on their amylose content into waxy (0-2%), very low (3-9%), low (10-19%), intermediate (20-25%), and high (> 25%) (Kongseree and Juliano, 1972). The most widely used method for amylose determination is a colorimetric assay where iodine binds with amylose to produce a blue-purple color, which is measured spectrophotometrically at a single wavelength (620nm). Low amylose content is usually associated with tender, cohesive and glossy cooked rice; while, high amylose content is associated with firm, fluffy and separate grains of cooked rice. The Waxy (Wx) gene, which encodes granule-bound starch synthase I (GBSSI), is the major gene controlling AC in rice (Nakamura, 2002). The Waxy gene is located on chromosome six and various single nucleotide polymorphisms (SNPs) of Wx were found, including a ‘G’ to ‘T’ SNP of the first intron, ‘A’ to ‘C’ SNP of the sixth exon and ‘C’ to ‘T’ SNP of the tenth exon (Larkin and Park, 2003). The ‘AGGTATA’ sequence at the 5’splice-junction coincides with the presence of the Wxa allele, while the ‘AGTTATA’ sequence coincides with the presence of the Wxb allele. Therefore, all intermediate and high amylose cultivars had ‘G’ nucleotide while low amylose cultivars had ‘T’ nucleotide at the putative leader intron 5′ splice site. The cytosine and thymidine (CT) dinucleotide repeats in the 5’- untranslated region (UTR) of the Waxy gene were reported to be a factor associated with AC. However, the relationship between these polymorphisms and amylose contents is not clear. Amylopectin chain length distribution plays a very important role to determine GT in cooked rice. The time required for cooking is determined by the gelatinization temperature of starch. It is important because it affects the texture of cooked rice and it is related to the cooking time of rice. The gelatinization temperature is estimated by the alkali digestibility test. It is measured by the alkali spreading value (ASV). The degree of spreading value of individual milled rice kernels in a weak alkali solution (1.7% KOH) is very closely correlated with gelatinized temperature. According to the ASV, rice varieties may be classified as low (55 to 69°C), intermediate (70 to 74°C) and high (> 74°C) GT classes. In a breeding program ASV is extensively used to estimate the gelatinization temperature. The synthesis of amylopectin is more complex than that of amylose. Polymorphisms in the starch synthase IIa (SSIIa) gene which is recognized as the Alk gene are responsible for the differences in GT in rice (Umemoto and Aoki, 2005; Waters et al., 2006). Two single nucleotide polymorphisms (SNPs) in the last exon of the Alk gene are responsible for the differences in GT in rice. The biochemical analysis clearly showed that the function of the amino acids caused by these two SNPs is essential for SSIIa enzyme activity (Nakamura et al., 2005) and those are ‘G’/‘A’ SNP at 4424 bp position and ‘GC’/‘TT’ SNPs at 4533/4534 bp position with reference to Nipponbare rice genomic sequence. Based on the SNPs, Low SSIIa enzyme activity results in S-type amylopectin, which is enriched in short chains whereas high SSIIa enzyme activity produces L-type amylopectin (Umemoto et al., 2004). Therefore, the combination of ‘G’ at SNP3 and ‘GC’ at SNP4 is required to produce L-type rice starch and this has a higher GT relative to S-type starch. GC is a standard assay that is used in rice improvement programs to determine the texture of softness and firmness in high amylose rice cultivars. Intermediate and low amylose rice usually has soft gel consistency. Sequence variation in exon 10 of the Waxy gene associates with GC (Tran et al., 2011).OBJECTIVES The objectives of this study were to detect polymorphisms in major starch synthesizing genes among several rice cultivars as models and to determine the relationship between their SNP variations and starch physicochemical properties. Also, we analyzed major starch synthesizing gene sequences of several Sri Lankan rice varieties in silico aiming at utilizing this information in rice breeding programs.MATERIALS AND METHODSPlant materials: Seeds of eight Oryza sativa L. accessions were obtained from the Rice Research and Development Institute (RRDI), Bathalagoda, Sri Lanka and Gene Bank of Plant Genetic Resource Center (PGRC), Gannoruwa.Characterization of grain physical parameters: Grain length and width were determined using a vernier caliper. Ten grains from each sample were collected randomly and measured to obtain the average length and width of the milled rice. The average length and width were recorded as their length and width. Based on the length and width of the grains, the milled rice grains were classified into four classes (table 1) according to the method accepted by RRDI Bathalagoda, Sri Lanka.According to the scale L/S – Long Slender, L/M – Long Medium, I/B – Intermediate Bold and S/R –Short RoundAnalysis of amylose content: Initially, rice samples were dehusked and polished prior to milling. Ten whole – milled rice kernels of eight rice samples were ground separately by using mortar and pestle. Amylose content per 100 mg was determined by measuring the blue value of rice varieties as described by Juliano (1971). About 100mg rice sample was shifted into a 100 mL volumetric flask and 1mL of 95% ethanol was added. Then 9mL of 1N NaOH was added and the content was boiled for 20min. at boiling temperature to gelatinize the starch. After cooling the content, the volume was made up to 100mL and 5mL of starch solution was pipetted out into a 100mL volumetric flask. The blue color was developed by adding 1mL of 1N acetic acid and 2 mL of iodine solution (0.2g iodine and 2.0g potassium iodine in 10 mL aqueous solution). Then volume was made up to 100mL with distilled water and the solution was kept for 20min. after shaking. Finally, the absorbance of the solution was measured at 620nm using Spectrophotometer T80 (PG Instruments Limited) as described by Juliano (1971). The standard curve was prepared using 40mg of potato-amylose to calculate the amylose content of rice varieties through absorbance values. Forty mg of potato amylose was put into a 100 mL of volumetric flask and 1ml of 95% ethanol and 9mL of NaOH were added and content was heated for 20min at boiling temperature. After cooling the content volume of the solution was made up to 100mL using distilled water. Then 1mL, 2mL, 3mL, 4mL and 5mL of amylose solution were pipetted out into 100mL flasks. Then 0.2mL, 0.4mL, 0.6mL, 0.8mL and 1mL of 1N acetic acid were added to the flasks respectively. Finally, 2mL of iodine solution was added to each flask and volume was made up to 100mL with distilled water. Solutions were stood up for 20min. after shaking and absorbance values were measured at 620nm. Measured absorbance values were plotted at 620nm against the concentration of anhydrous amylose (mg).Analysis of gelatinization temperature: GT was indirectly measured on rice by the alkali spreading value. Husked and polished seeds per accession were used for the analysis. Selected duplicate sets of six milled grains without cracks of each sample were put into Petri dishes. About 10mL of 1.7% KOH was added and grains were spread in the petri dish to provide enough space. The constant temperature at 30°C was maintained to ensure better reproducibility. After 23hrs, the degree of disintegration was quantified by a standard protocol with a numerical scale of 1–7 (table 2) as reported by Cruz and Khush (2000). As reported by Juliano (2003), GT of rice was determined using the alkaline spreading scale, where 1.0-2.5: High (74-80 °C), 2.6-3.4: High-intermediate (70-74 °C), 3.5-5.4: Intermediate (70-74 °C) and 5.5-7.0 Low: (55-70 °C).Bioinformatics and statistical analysis: The available literature was used to identify the most likely candidate genes associated with rice starch quality and their SNPs of each gene (Hirose et al., 2006; Waters and Henry, 2007; Tran et al., 2011). In all the tested varieties except Bg 352 and At 354, the DNA sequence of each gene was retrieved from the Rice SNP Seek database (http://snp-seek.irri.org/). The gene sequences of At 354 and Bg 352 were obtained from the National Research Council 16-016 project, Wayamba University of Sri Lanka. Multiple sequence alignment was conducted for the DNA sequence using Clustal Omegavsoftware (https://www.ebi.ac.uk/Tools/msa/clustalo/). Starch physiochemical data obtained were subjected to a one-way analysis of variance (ANOVA) followed by Duncan’s New Multiple Range Test (DNMRT) to determine the statistical differences among varieties at the significance level of p ≤ 0.05. Statistical analysis was done using SAS version 9.1 (SAS, 2004).ESULTS AND DISCUSSION: Physical properties of rice grains: Physical properties such as length, width, size, shape and pericarp color of rice grains obtained from eight different rice varieties are given in table 3. Classification of rice grains was carried out, according to their sizes and shapes based on Juliano (1985). The size of the rice grains was determined as per grain length while grain shape was determined by means of length and width ratio of the rice kernel. In the local market, rice is classified as Samba (short grain), Nadu (intermediate grain) and Kora (long/medium) based on the size of the grain (Pathiraje et al., 2010). Lengths of rice kernels were varied from 5.58 to 6.725 mm for all varieties. The highest grain length and width were given by At 354 and Pachchaperumal respectively. The varieties, Bw 295-5 and H 6 showed a length: width ratio over 3 which is considered as slender in grain shape. Bw 295-5, H 6, At 354, Bg 352 and Nipponbare possessed white pericarp and others possessed red pericarp.Relationship between amylose content and SNPs variation of waxy loci in selected varieties: Amylose content was measured in seven Sri Lankan rice varieties and one exotic rice variety. Amylose content of the evaluated varieties varied significantly with p ≤ 0.05 with the lowest of 15.11% and highest of 28.63% which were found in Nipponbare and Bw 295-5, respectively (table 4). The majority of the evaluated varieties fell into the high AC category (between 25-28%). Only Nipponbare could be clearly categorized under the low amylose group (table 4). The amylose content of Bg 352, Pachchaperumal and Herathbanda have already been determined by early studies of Rebeira et al. (2014) and Fernando et al. (2015). Most of the data obtained in the present experiment has agreed with the results of previous studies. Major genes such as Waxy and their functional SNPs have a major influence on amylose in rice (Nakamura et al., 2005). Accordingly, single nucleotide polymorphism, ‘G’/‘T’, at the 5’ leader intron splice site of the GBSSI has explained the variation in amylose content of varieties. Accordingly, high and intermediate amylose varieties have ‘AGGTATA’ while low amylose varieties have the sequence ‘AGTTATA’, which might lead to a decrease in the splicing efficiency. Therefore, the GBSSI activity of Nipponbare might be considerably weak and resulted in starch with low amylose content. Hence, producing ‘G’/‘T’ polymorphism clearly differentiates low amylose rice varieties, as reported by Nakamura et al. (2005). In GBSSI, Larkin and Park (2003) identified an ‘A’/‘C’ polymorphism in exon 6 and a ‘C’/‘T’ polymorphism in exon 10 which resulted in non- synonymous amino acid change. Chen et al. (2008) reported that the non-synonymous ‘A’/‘C’ SNP at exon 6 had the highest possible impact on GBSSI. Accordingly, the ‘A’/‘C’ polymorphism in exon 6 causes a tyrosine/serine amino acid substitution while the ‘C’/‘T’ polymorphism in exon 10 causes a serine/proline amino acid substitution. In view of this information, there is a relationship between the polymorphism detected by in silico analysis and amylose content obtained from our experiment. Out of the eight tested rice varieties, only one variety, Nipponbare was categorized as low amylose variety (10-19%) and it exhibited ‘T’ nucleotide at the intron splice site (table 4; figure 1). Varieties such as Pachchaperumal, Balasuriya, Bw 295-5, H 6, Herathbanda, At 354 and Bg 352 which contained high amylose (> 25%), had ‘G’ and ‘A’ nucleotides at intron splice site and exon 6 respectively (table 4; figure 1). The predominant allelic pattern of intron splice site and exon 6 are different in varieties containing intermediate amylose content (20-25%) which showed ‘G’ and ‘C’ nucleotides respectively. Of these selected rice varieties, none of the intermediate type amylose variety was found.Relationship between gel consistency and SNPs variation in Waxy loci: In this study, GC data of Herathbanda, Hondarawalu, Kuruluthuda, Pachchaperumal and Bg 352 were obtained from Fernando et al. (2015). The results of Tran et al. (2011) showed that the exon 10 ‘C’/‘T’ SNP of Wx has mainly affected GC. Accordingly, rice with a ‘C’ at exon 10 had soft and viscous gels once cooked. However, a sample with a ‘T’ had short and firm gels. In this study, Herathbanda, Hondarawalu, Kuruluthuda and Pachchaperumal had ‘C’ nucleotide and Bg 352 had ‘T’ nucleotide in exon 10 (table 5; figure 2). However, ‘C’/‘T’ substitution analysis could not be used to explain the GC of tested varieties.Relationship between gelatinized temperature and SNPs variation of Alk loci in selected rice varieties: Although there were differences in the scores, the degree of disintegration of all samples was saturated at 23 hrs. Most of the selected rice varieties showed the intermediate disintegration score. Varieties, Pachchaperumal, Balasuriya, H 6, Herathbanda, At 354 and Bg 352 were categorized into intermediate GT class (70–74°C) as indicated by an alkali spreading (AS) value of 5 (table 6; figure 3). Nipponbare and Bw 295-5 showed the highest disintegration score indicating the dispersion of all grains. Hence these varieties were categorized into low GT class (55-69°C) as indicated by an AS value of 6 (table 6; figure 3). However, high GT class rice varieties (> 74°C) were not found in the tested samples. Chromosomal mutation within the Alk gene has led to a number of single nucleotide polymorphisms (SNPs). Umemoto et al. (2004) identified four SNPs in Alk gene. Thus, SNP3 and SNP4 may be important genetic polymorphisms that are associated with GT class. According to the SNP3 and SNP4, eight rice varieties could be classified into either high GT or low GT types. If there is ‘A’ instead of ‘G’ at 4424 bp position of Alk gene with reference to Nipponbare rice genomic sequence, it codes methionine instead of valine amino acid residue in SSIIa, whilst two adjacent SNPs at bases 4533 and 4534 code for either leucine (‘GC’) or phenylalanine (‘TT’). Rice varieties with high GT starch had a combination of valine and leucine at these residues. Rice varieties with low GT starch had a combination of either methionine and leucine or valine and phenylalanine at these same residues. Nipponbare carried the ‘A’ and ‘GC’ nucleotides, while Bw 295-5 carried the ‘G’ and ‘TT’ nucleotides. Hence these varieties were classified into low GT class. Varieties such as Pachchaperumal, Balasuriya, H 6, Herathbanda, At 354 and Bg 352 carried ‘G’ and ‘GC’ nucleotides and these varieties were classified into high GT rice varieties. However, intermediate GT status could not be determined by SNP3 and SNP4 mutation of Alk gene (table 6; figure 4).In silico analysis of the polymorphisms in GBSSI gene and Alk genes of rice varieties retrieved from Rice-SNP-database: In this study, GBSSI gene and Alk gene were compared with the sequences retrieved from the Rice-SNP-Seek database to validate the SNPs further. As previously reported by Ayres et al. (1997), all low amylose varieties had the sequence ‘AGTTATA’ in exon 1. In agreement with preliminary work done by Larkin and Park (2003), all of the intermediate amylose varieties have the allelic pattern of GCC. All of the high amylose varieties have either the GAC or GAT allele of GBSSI. Among 42 rice accessions with the Sri Lankan pedigree, four allelic patterns were found; TAC, GCC, GAC and GAT (table 7). In this allelic pattern, the first letter corresponds to the ‘G’/‘T’ polymorphism in 5’ leader intron splice-junction, the second letter corresponds to the ‘A’/‘C’ polymorphism in exon 6 and the third letter corresponds to the ‘C’/‘T’ polymorphism in exon10 of Waxy gene. Analysis of the ‘G’/‘T’ polymorphism in the Wx locus showed that 41 rice cultivars shared the same ‘AGGTATA’ sequence at the 5’ leader intron splice-junction. But only 1 rice cultivar, Puttu nellu was found with ‘T’ nucleotide in intron1/exon1 junction site, which could be categorized as a low amylose variety (table 7). As discussed above, varieties with an intermediate level of apparent amylose could be reliably distinguished from those with higher apparent amylose based on a SNP in exon 6. Hence, only three rice varieties Nalumoolai Karuppan, Pannithi and Godawel with ‘C’ nucleotide in exon 6 exhibited the possibility of containing intermediate amylose content (table 7). High activity of GBSSI produces high amylose content leading to a non-waxy, non-sticky or non-glutinous phenotype. Therefore, according to the in silico genotypic results, rest of the 38 rice varieties may produce high amylose content in the endosperm (table 7). Proving this phenomenon. Abeysekera et al. (2017) has reported that usually, most of Sri Lankan rice varieties contain high amylose content. Targeted sequence analysis of exon 8 of the Alk gene in 42 different rice cultivars were found with three SNP polymorphisms that resulted in a changed amino acid sequence and, of these three SNPs, two SNPs were reported to be correlated with possible GT differences. Accordingly, Puttu nellu and 3210 rice varieties carried the ‘G’ and ‘TT’ nucleotides in SNP3 and SNP4 respectively (table 7). Hence these varieties can be classified into low GT class and except these two; other rice varieties carried the ‘G’ and ‘GC’ nucleotides in SNP3 and SNP4 respectively. Therefore, those varieties can possibly be classified into high GT rice varieties (table 7). However, further experiments are necessary to check the phenotypic variations for grain amylose content and GT class of in silico analyzed rice varieties. CONCLUSION Present results revealed the relationship between SNPs variation at Waxy loci and the amylose content of selected rice varieties. Accordingly, Pachchaperumal, At 354, Bg 352, Herathbanda, H 6, Balasuriya and Bw 295-5 with high amylose content had ‘G’ instead of ‘T’ in the first intron exhibiting the presence of Wxa allele with reference to Nipponbare which had low amylose content. Also all tested varieties had ‘A’ in exon 6 of the Waxy gene. Thus present findings i.e. presence of Wxa allele and SNP ‘A’ in exon 6 could be used as a potential molecular marker for the selection of high amylose varieties. In addition, Bw 295-5 which is a low GT variety, had two SNPs variations in the last exon of the Alk gene i.e. ‘G’ and ‘TT’ which is likely to be used to represent low GT class. Accordingly, sequence variations identified in Waxy and Alk genes could be utilized in the future rice breeding programs for the development of varieties with preferential amylose content and GT class.ACKNOWLEDGMENTSDirector and staff of the Gene Bank, Plant Genetic Resources Center, Gannoruwa are acknowledged for giving rice accessions.CONFLICT OF INTERESTAuthors have no conflict of interest.REFERENCESAbeysekera, W., G. Premakumara, A. Bentota and D. S. Abeysiriwardena, 2017. Grain amylose content and its stability over seasons in a selected set of rice varieties grown in Sri Lanka. Journal of agricultural sciences Sri Lanka, 12(1): 43-50.Ayres, N., A. McClung, P. Larkin, H. Bligh, C. Jones and W. Park, 1997. Microsatellites and a single-nucleotide polymorphism differentiate apparentamylose classes in an extended pedigree of us rice germ plasm. Theoretical applied genetics, 94(6-7): 773-781.Chen, M.-H., C. Bergman, S. Pinson and R. Fjellstrom, 2008. Waxy gene haplotypes: Associations with apparent amylose content and the effect by the environment in an international rice germplasm collection. Journal of cereal science, 47(3): 536-545.Cruz, N. D. and G. Khush, 2000. Rice grain quality evaluation procedures. Aromatic rices, 3: 15-28.Fernando, H., T. Kajenthini, S. Rebeira, T. Bamunuarachchige and H. Wickramasinghe, 2015. Validation of molecular markers for the analysis of genetic diversity of amylase content and gel consistency among representative rice varieties in sri lanka. Tropical agricultural research, 26(2): 317-328.Hirose, T., T. Ohdan, Y. Nakamura and T. Terao, 2006. Expression profiling of genes related to starch synthesis in rice leaf sheaths during the heading period. Physiologia plantarum, 128(3): 425-435.Juliano, B., 1971. A simplified assay for milled rice amylose. Journal of cereal science today, 16: 334-360.Juliano, B. O., 1985. Rice: Chemistry and technology. The american association of cereal chemists. Inc. St. Paul, Minnesota, USA, 774.Juliano, B. O., 2003. Rice chemistry and quality. Island publishing house. Island publishing house, Manila: 1-7.Kharabian-Masouleh, A., D. L. Waters, R. F. Reinke, R. Ward and R. J. Henry, 2012. Snp in starch biosynthesis genes associated with nutritional and functional properties of rice. Scientific reports, 2(1): 1-9.Kongseree, N. and B. O. Juliano, 1972. Physicochemical properties of rice grain and starch from lines differing in amylose content and gelatinization temperature. Journal of agricultural food chemistry, 20(3): 714-718.Larkin, P. D. and W. D. Park, 2003. Association of waxy gene single nucleotide polymorphisms with starch characteristics in rice (Oryza sativa L.). Molecular Breeding, 12(4): 335-339.Nakamura, Y., 2002. Towards a better understanding of the metabolic system for amylopectin biosynthesis in plants: Rice endosperm as a model tissue. Plant cell physiology, 43(7): 718-725.Nakamura, Y., P. B. Francisco, Y. Hosaka, A. Sato, T. Sawada, A. Kubo and N. Fujita, 2005. Essential amino acids of starch synthase iia differentiate amylopectin structure and starch quality between Japonica and Indica rice varieties. Plant molecular biology, 58(2): 213-227.Pathiraje, P., W. Madhujith, A. Chandrasekara and S. Nissanka, 2010. The effect of rice variety and parboiling on in vivo glycemic response. Journal of tropical agricultural research, 22(1): 26-33.Rebeira, S., H. Wickramasinghe, W. Samarasinghe and B. Prashantha, 2014. Diversity of grain quality characteristics of traditional rice (Oryza sativa L.) varieties in sri lanka. Tropical agricultural research, 25(4): 470-478.Sartaj, I. Z. and S. A. E. R. Suraweera, 2005. Comparison of different parboiling methods on the quality characteristics of rice. Annals of the Sri Lankan Department of Agriculture, 7: 245-252.Tran, N., V. Daygon, A. Resurreccion, R. Cuevas, H. Corpuz and M. Fitzgerald, 2011. A single nucleotide polymorphism in the waxy gene explains a significant component of gel consistency. Theoretical applied genetics, 123(4): 519-525.Umemoto, T. and N. Aoki, 2005. Single-nucleotide polymorphisms in rice starch synthase iia that alter starch gelatinisation and starch association of the enzyme. Functional plant biology, 32(9): 763-768.Umemoto, T., N. Aoki, H. Lin, Y. Nakamura, N. Inouchi, Y. Sato, M. Yano, H. Hirabayashi and S. Maruyama, 2004. Natural variation in rice starch synthase iia affects enzyme and starch properties. Functional plant biology, 31(7): 671-684.Waters, D. L. and R. J. Henry, 2007. Genetic manipulation of starch properties in plants: Patents 2001-2006. Recent patents on biotechnology, 1(3): 252-259.Waters, D. L., R. J. Henry, R. F. Reinke and M. A. Fitzgerald, 2006. Gelatinization temperature of rice explained by polymorphisms in starch synthase. Plant biotechnology journal, 4(1): 115-122.

To date, discrete event stochastic simulations of large scale biological reaction systems are extremely compute-intensive and time-consuming. Besides, it has been widely accepted that spatial factor plays a critical role in the dynamics of most biological reaction systems. The NSM (the Next Sub-Volume Method), a spatial variation of the Gillespie’s stochastic simulation algorithm (SSA), has been proposed for spatially stochastic simulation of those systems. While being able to explore high degree of parallelism in systems, NSM is inherently sequential, which still suffers from the problem of low simulation speed. Fine-grained parallel execution is an elegant way to speed up sequential simulations. Thus, based on the discrete event simulation framework JAMES II, we design and implement a PDES (Parallel Discrete Event Simulation) TW (time warp) simulator to enable the fine-grained parallel execution of spatial stochastic simulations of biological reaction systems using the ANSM (the Abstract NSM), a parallel variation of the NSM. The simulation results of classical Lotka-Volterra biological reaction system show that our time warp simulator obtains remarkable parallel speed-up against sequential execution of the NSM.I.IntroductionThe goal of Systems biology is to obtain system-level investigations of the structure and behavior of biological reaction systems by integrating biology with system theory, mathematics and computer science [1][3], since the isolated knowledge of parts can not explain the dynamics of a whole system. As the complement of “wet-lab” experiments, stochastic simulation, being called the “dry-computational” experiment, plays a more and more important role in computing systems biology [2]. Among many methods explored in systems biology, discrete event stochastic simulation is of greatly importance [4][5][6], since a great number of researches have present that stochasticity or “noise” have a crucial effect on the dynamics of small population biological reaction systems [4][7]. Furthermore, recent research shows that the stochasticity is not only important in biological reaction systems with small population but also in some moderate/large population systems [7].To date, Gillespie’s SSA [8] is widely considered to be the most accurate way to capture the dynamics of biological reaction systems instead of traditional mathematical method [5][9]. However, SSA-based stochastic simulation is confronted with two main challenges: Firstly, this type of simulation is extremely time-consuming, since when the types of species and the number of reactions in the biological system are large, SSA requires a huge amount of steps to sample these reactions; Secondly, the assumption that the systems are spatially homogeneous or well-stirred is hardly met in most real biological systems and spatial factors play a key role in the behaviors of most real biological systems [19][20][21][22][23][24]. The next sub-volume method (NSM) [18], presents us an elegant way to access the special problem via domain partition. To our disappointment, sequential stochastic simulation with the NSM is still very time-consuming, and additionally introduced diffusion among neighbor sub-volumes makes things worse. Whereas, the NSM explores a very high degree of parallelism among sub-volumes, and parallelization has been widely accepted as the most meaningful way to tackle the performance bottleneck of sequential simulations [26][27]. Thus, adapting parallel discrete event simulation (PDES) techniques to discrete event stochastic simulation would be particularly promising. Although there are a few attempts have been conducted [29][30][31], research in this filed is still in its infancy and many issues are in need of further discussion. The next section of the paper presents the background and related work in this domain. In section III, we give the details of design and implementation of model interfaces of LP paradigm and the time warp simulator based on the discrete event simulation framework JAMES II; the benchmark model and experiment results are shown in Section IV; in the last section, we conclude the paper with some future work.II. Background and Related WorkA. Parallel Discrete Event Simulation (PDES)The notion Logical Process (LP) is introduced to PDES as the abstract of the physical process [26], where a system consisting of many physical processes is usually modeled by a set of LP. LP is regarded as the smallest unit that can be executed in PDES and each LP holds a sub-partition of the whole system’s state variables as its private ones. When a LP processes an event, it can only modify the state variables of its own. If one LP needs to modify one of its neighbors’ state variables, it has to schedule an event to the target neighbor. That is to say event message exchanging is the only way that LPs interact with each other. Because of the data dependences or interactions among LPs, synchronization protocols have to be introduced to PDES to guarantee the so-called local causality constraint (LCC) [26]. By now, there are a larger number of synchronization algorithms have been proposed, e.g. the null-message [26], the time warp (TW) [32], breath time warp (BTW) [33] and etc. According to whether can events of LPs be processed optimistically, they are generally divided into two types: conservative algorithms and optimistic algorithms. However, Dematté and Mazza have theoretically pointed out the disadvantages of pure conservative parallel simulation for biochemical reaction systems [31]. B. NSM and ANSM The NSM is a spatial variation of Gillespie’ SSA, which integrates the direct method (DM) [8] with the next reaction method (NRM) [25]. The NSM presents us a pretty good way to tackle the aspect of space in biological systems by partitioning a spatially inhomogeneous system into many much more smaller “homogeneous” ones, which can be simulated by SSA separately. However, the NSM is inherently combined with the sequential semantics, and all sub-volumes share one common data structure for events or messages. Thus, directly parallelization of the NSM may be confronted with the so-called boundary problem and high costs of synchronously accessing the common data structure [29]. In order to obtain higher efficiency of parallel simulation, parallelization of NSM has to firstly free the NSM from the sequential semantics and secondly partition the shared data structure into many “parallel” ones. One of these is the abstract next sub-volume method (ANSM) [30]. In the ANSM, each sub-volume is modeled by a logical process (LP) based on the LP paradigm of PDES, where each LP held its own event queue and state variables (see Fig. 1). In addition, the so-called retraction mechanism was introduced in the ANSM too (see algorithm 1). Besides, based on the ANSM, Wang etc. [30] have experimentally tested the performance of several PDES algorithms in the platform called YH-SUPE [27]. However, their platform is designed for general simulation applications, thus it would sacrifice some performance for being not able to take into account the characteristics of biological reaction systems. Using the similar ideas of the ANSM, Dematté and Mazza have designed and realized an optimistic simulator. However, they processed events in time-stepped manner, which would lose a specific degree of precisions compared with the discrete event manner, and it is very hard to transfer a time-stepped simulation to a discrete event one. In addition, Jeschke etc.[29] have designed and implemented a dynamic time-window simulator to execution the NSM in parallel on the grid computing environment, however, they paid main attention on the analysis of communication costs and determining a better size of the time-window.Fig. 1: the variations from SSA to NSM and from NSM to ANSMC. JAMES II JAMES II is an open source discrete event simulation experiment framework developed by the University of Rostock in Germany. It focuses on high flexibility and scalability [11][13]. Based on the plug-in scheme [12], each function of JAMES II is defined as a specific plug-in type, and all plug-in types and plug-ins are declared in XML-files [13]. Combined with the factory method pattern JAMES II innovatively split up the model and simulator, which makes JAMES II is very flexible to add and reuse both of models and simulators. In addition, JAMES II supports various types of modelling formalisms, e.g. cellular automata, discrete event system specification (DEVS), SpacePi, StochasticPi and etc.[14]. Besides, a well-defined simulator selection mechanism is designed and developed in JAMES II, which can not only automatically choose the proper simulators according to the modeling formalism but also pick out a specific simulator from a serious of simulators supporting the same modeling formalism according to the user settings [15].III. The Model Interface and SimulatorAs we have mentioned in section II (part C), model and simulator are split up into two separate parts. Thus, in this section, we introduce the designation and implementation of model interface of LP paradigm and more importantly the time warp simulator.A. The Mod Interface of LP ParadigmJAMES II provides abstract model interfaces for different modeling formalism, based on which Wang etc. have designed and implemented model interface of LP paradigm[16]. However, this interface is not scalable well for parallel and distributed simulation of larger scale systems. In our implementation, we accommodate the interface to the situation of parallel and distributed situations. Firstly, the neighbor LP’s reference is replaced by its name in LP’s neighbor queue, because it is improper even dangerous that a local LP hold the references of other LPs in remote memory space. In addition, (pseudo-)random number plays a crucial role to obtain valid and meaningful results in stochastic simulations. However, it is still a very challenge work to find a good random number generator (RNG) [34]. Thus, in order to focus on our problems, we introduce one of the uniform RNGs of JAMES II to this model interface, where each LP holds a private RNG so that random number streams of different LPs can be independent stochastically. B. The Time Warp SimulatorBased on the simulator interface provided by JAMES II, we design and implement the time warp simulator, which contains the (master-)simulator, (LP-)simulator. The simulator works strictly as master/worker(s) paradigm for fine-grained parallel and distributed stochastic simulations. Communication costs are crucial to the performance of a fine-grained parallel and distributed simulation. Based on the Java remote method invocation (RMI) mechanism, P2P (peer-to-peer) communication is implemented among all (master-and LP-)simulators, where a simulator holds all the proxies of targeted ones that work on remote workers. One of the advantages of this communication approach is that PDES codes can be transferred to various hardwire environment, such as Clusters, Grids and distributed computing environment, with only a little modification; The other is that RMI mechanism is easy to realized and independent to any other non-Java libraries. Since the straggler event problem, states have to be saved to rollback events that are pre-processed optimistically. Each time being modified, the state is cloned to a queue by Java clone mechanism. Problem of this copy state saving approach is that it would cause loads of memory space. However, the problem can be made up by a condign GVT calculating mechanism. GVT reduction scheme also has a significant impact on the performance of parallel simulators, since it marks the highest time boundary of events that can be committed so that memories of fossils (processed events and states) less than GVT can be reallocated. GVT calculating is a very knotty for the notorious simultaneous reporting problem and transient messages problem. According to our problem, another GVT algorithm, called Twice Notification (TN-GVT) (see algorithm 2), is contributed to this already rich repository instead of implementing one of GVT algorithms in reference [26] and [28].This algorithm looks like the synchronous algorithm described in reference [26] (pp. 114), however, they are essentially different from each other. This algorithm has never stopped the simulators from processing events when GVT reduction, while algorithm in reference [26] blocks all simulators for GVT calculating. As for the transient message problem, it can be neglect in our implementation, because RMI based remote communication approach is synchronized, that means a simulator will not go on its processing until the remote the massage get to its destination. And because of this, the high-costs message acknowledgement, prevalent over many classical asynchronous GVT algorithms, is not needed anymore too, which should be constructive to the whole performance of the time warp simulator.IV. Benchmark Model and Experiment ResultsA. The Lotka-Volterra Predator-prey SystemIn our experiment, the spatial version of Lotka-Volterra predator-prey system is introduced as the benchmark model (see Fig. 2). We choose the system for two considerations: 1) this system is a classical experimental model that has been used in many related researches [8][30][31], so it is credible and the simulation results are comparable; 2) it is simple but helpful enough to test the issues we are interested in. The space of predator-prey System is partitioned into a2D NXNgrid, whereNdenotes the edge size of the grid. Initially the population of the Grass, Preys and Predators are set to 1000 in each single sub-volume (LP). In Fig. 2,r1,r2,r3stand for the reaction constants of the reaction 1, 2 and 3 respectively. We usedGrass,dPreyanddPredatorto stand for the diffusion rate of Grass, Prey and Predator separately. Being similar to reference [8], we also take the assumption that the population of the grass remains stable, and thusdGrassis set to zero.R1:Grass + Prey ->2Prey(1)R2:Predator +Prey -> 2Predator(2)R3:Predator -> NULL(3)r1=0.01; r2=0.01; r3=10(4)dGrass=0.0;dPrey=2.5;dPredato=5.0(5)Fig. 2: predator-prey systemB. Experiment ResultsThe simulation runs have been executed on a Linux Cluster with 40 computing nodes. Each computing node is equipped with two 64bit 2.53 GHz Intel Xeon QuadCore Processors with 24GB RAM, and nodes are interconnected with Gigabit Ethernet connection. The operating system is Kylin Server 3.5, with kernel 2.6.18. Experiments have been conducted on the benchmark model of different size of mode to investigate the execution time and speedup of the time warp simulator. As shown in Fig. 3, the execution time of simulation on single processor with 8 cores is compared. The result shows that it will take more wall clock time to simulate much larger scale systems for the same simulation time. This testifies the fact that larger scale systems will leads to more events in the same time interval. More importantly, the blue line shows that the sequential simulation performance declines very fast when the mode scale becomes large. The bottleneck of sequential simulator is due to the costs of accessing a long event queue to choose the next events. Besides, from the comparison between group 1 and group 2 in this experiment, we could also conclude that high diffusion rate increased the simulation time greatly both in sequential and parallel simulations. This is because LP paradigm has to split diffusion into two processes (diffusion (in) and diffusion (out) event) for two interactive LPs involved in diffusion and high diffusion rate will lead to high proportional of diffusion to reaction. In the second step shown in Fig. 4, the relationship between the speedups from time warp of two different model sizes and the number of work cores involved are demonstrated. The speedup is calculated against the sequential execution of the spatial reaction-diffusion systems model with the same model size and parameters using NSM.Fig. 4 shows the comparison of speedup of time warp on a64X64grid and a100X100grid. In the case of a64X64grid, under the condition that only one node is used, the lowest speedup (a little bigger than 1) is achieved when two cores involved, and the highest speedup (about 6) is achieved when 8 cores involved. The influence of the number of cores used in parallel simulation is investigated. In most cases, large number of cores could bring in considerable improvements in the performance of parallel simulation. Also, compared with the two results in Fig. 4, the simulation of larger model achieves better speedup. Combined with time tests (Fig. 3), we find that sequential simulator’s performance declines sharply when the model scale becomes very large, which makes the time warp simulator get better speed-up correspondingly.Fig. 3: Execution time (wall clock time) of Seq. and time warp with respect to different model sizes (N=32, 64, 100, and 128) and model parameters based on single computing node with 8 cores. Results of the test are grouped by the diffusion rates (Group 1: Sequential 1 and Time Warp 1. dPrey=2.5, dPredator=5.0; Group 2: dPrey=0.25, dPredator=0.5, Sequential 2 and Time Warp 2).Fig. 4: Speedup of time warp with respect to the number of work cores and the model size (N=64 and 100). Work cores are chose from one computing node. Diffusion rates are dPrey=2.5, dPredator=5.0 and dGrass=0.0.V. Conclusion and Future WorkIn this paper, a time warp simulator based on the discrete event simulation framework JAMES II is designed and implemented for fine-grained parallel and distributed discrete event spatial stochastic simulation of biological reaction systems. Several challenges have been overcome, such as state saving, roll back and especially GVT reduction in parallel execution of simulations. The Lotka-Volterra Predator-Prey system is chosen as the benchmark model to test the performance of our time warp simulator and the best experiment results show that it can obtain about 6 times of speed-up against the sequential simulation. The domain this paper concerns with is in the infancy, many interesting issues are worthy of further investigated, e.g. there are many excellent PDES optimistic synchronization algorithms (e.g. the BTW) as well. Next step, we would like to fill some of them into JAMES II. In addition, Gillespie approximation methods (tau-leap[10] etc.) sacrifice some degree of precision for higher simulation speed, but still could not address the aspect of space of biological reaction systems. The combination of spatial element and approximation methods would be very interesting and promising; however, the parallel execution of tau-leap methods should have to overcome many obstacles on the road ahead.AcknowledgmentThis work is supported by the National Natural Science Foundation of China (NSF) Grant (No.60773019) and the Ph.D. Programs Foundation of Ministry of Education of China (No. 200899980004). The authors would like to show their great gratitude to Dr. Jan Himmelspach and Dr. Roland Ewald at the University of Rostock, Germany for their invaluable advice and kindly help with JAMES II.ReferencesH. Kitano, "Computational systems biology." Nature, vol. 420, no. 6912, pp. 206-210, November 2002.H. Kitano, "Systems biology: a brief overview." 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We’ll have trouble now!The Archaeological Society of Jutland was founded on Sunday, 11 March 1951. As with most projects with which P.V Glob was involved, this did not pass off without drama. Museum people and amateur archaeologists in large numbers appeared at the Museum of Natural History in Aarhus, which had placed rooms at our disposal. The notable dentist Holger Friis, the uncrowned king of Hjørring, was present, as was Dr Balslev from Aidt, Mr and Mrs Overgaard from Holstebro Museum, and the temperamental leader of Aalborg Historical Museum, Peter Riismøller, with a number of his disciples. The staff of the newly-founded Prehistoric Museum functioned as the hosts, except that one of them was missing: the instigator of the whole enterprise, Mr Glob. As the time for the meeting approached, a cold sweat broke out on the foreheads of the people present. Finally, just one minute before the meeting was to start, he arrived and mounted the platform. Everything then went as expected. An executive committee was elected after some discussion, laws were passed, and then suddenly Glob vanished again, only to materialise later in the museum, where he confided to us that his family, which included four children, had been enlarged by a daughter.That’s how the society was founded, and there is not much to add about this. However, a few words concerning the background of the society and its place in a larger context may be appropriate. A small piece of museum history is about to be unfolded.The story begins at the National Museum in the years immediately after World War II, at a time when the German occupation and its incidents were still terribly fresh in everyone’s memory. Therkel Mathiassen was managing what was then called the First Department, which covered the prehistoric periods.Although not sparkling with humour, he was a reliable and benevolent person. Number two in the order of precedence was Hans Christian Broholm, a more colourful personality – awesome as he walked down the corridors, with his massive proportions and a voice that sounded like thunder when nothing seemed to be going his way, as quite often seemed to be the case. Glob, a relatively new museum keeper, was also quite loud at times – his hot-blooded artist’s nature manifested itself in peculiar ways, but his straight forward appearance made him popular with both the older and the younger generations. His somewhat younger colleague C.J. Becker was a scholar to his fingertips, and he sometimes acted as a welcome counterbalance to Glob. At the bottom of the hierarchy was the student group, to which I belonged. The older students handled various tasks, including periodic excavations. This was paid work, and although the salary was by no means princely, it did keep us alive. Student grants were non-existent at the time. Four of us made up a team: Olfert Voss, Mogens Ørsnes, Georg Kunwald and myself. Like young people in general, we were highly discontented with the way our profession was being run by its ”ruling” members, and we were full of ideas for improvement, some of which have later been – or are being – introduced.At the top of our wish list was a central register, of which Voss was the strongest advocate. During the well over one hundred years that archaeology had existed as a professional discipline, the number of artefacts had grown to enormous amounts. The picture was even worse if the collections of the provincial museums were taken into consideration. We imagined how it all could be registered in a card index and categorised according to groups to facilitate access to references in any particular situation. Electronic data processing was still unheard of in those days, but since the introduction of computers, such a comprehensive record has become more feasible.We were also sceptical of the excavation techniques used at the time – they were basically adequate, but they badly needed tightening up. As I mentioned before, we were often working in the field, and not just doing minor jobs but also more important tasks, so we had every opportunity to try out our ideas. Kunwald was the driving force in this respect, working with details, using sections – then a novelty – and proceeding as he did with a thoroughness that even his fellow students found a bit exaggerated at times, although we agreed with his principles. Therkel Mathiassen moaned that we youngsters were too expensive, but he put up with our excesses and so must have found us somewhat valuable. Very valuable indeed to everyon e was Ejnar Dyggve’s excavation of the Jelling mounds in the early 1940s. From a Danish point of view, it was way ahead of its time.Therkel Mathiassen justly complained about the economic situation of the National Museum. Following the German occupation, the country was impoverished and very little money was available for archaeological research: the total sum available for the year 1949 was 20,000 DKK, which corresponded to the annual income of a wealthy man, and was of course absolutely inadequate. Of course our small debating society wanted this sum to be increased, and for once we didn’t leave it at the theoretical level.Voss was lucky enough to know a member of the Folketing (parliament), and a party leader at that. He was brought into the picture, and between us we came up with a plan. An article was written – ”Preserve your heritage” (a quotation from Johannes V. Jensen’s Denmark Song) – which was sent to the newspaper Information. It was published, and with a little help on our part the rest of the media, including radio, picked up the story.We informed our superiors only at the last minute, when everything was arranged. They were taken by surprise but played their parts well, as expected, and everything went according to plan. The result was a considerable increase in excavation funds the following year.It should be added that our reform plans included the conduct of exhibitions. We found the traditional way of presenting the artefacts lined up in rows and series dull and outdated. However, we were not able to experiment within this field.Our visions expressed the natural collision with the established ways that comes with every new generation – almost as a law of nature, but most strongly when the time is ripe. And this was just after the war, when communication with foreign colleagues, having been discontinued for some years, was slowly picking up again. The Archaeological Society of Jutland was also a part of all this, so let us turn to what Hans Christian Andersen somewhat provocatively calls the ”main country”.Until 1949, only the University of Copenhagen provided a degree in prehistoric archaeology. However, in this year, the University of Aarhus founded a chair of archaeology, mainly at the instigation of the Lord Mayor, Svend Unmack Larsen, who was very in terested in archaeology. Glob applied for the position and obtained it, which encompassed responsibility for the old Aarhus Museum or, as it was to be renamed, the Prehistoric Museum (now Moesgaard Museum).These were landmark events to Glob – and to me, as it turned out. We had been working together for a number of years on the excavation of Galgebakken (”Callows Hill”) near Slots Bjergby, Glob as the excavation leader, and I as his assistant. He now offered me the job of museum curator at his new institution. This was somewhat surprising as I had not yet finished my education. The idea was that I was to finish my studies in remote Jutland – a plan that had to be given up rather quickly, though, for reasons which I will describe in the following. At the same time, Gunner Lange-Kornbak – also hand-picked from the National Museum – took up his office as a conservation officer.The three of us made up the permanent museum staff, quickly supplemented by Geoffrey Bibby, who turned out to be an invaluable colleague. He was English and had been stationed in the Faeroe Islands during the war, where he learned to speak Danish. After 1945 he worked for some years for an oil company in the Gulf of Persia, but after marrying Vibeke, he settled in her home town of Aarhus. As his academic background had involved prehistoric cultures he wanted to collaborate with the museum, which Glob readily permitted.This small initial flock governed by Glob was not permitted to indulge inidleness. Glob was a dynamic character, full of good and not so good ideas, but also possessing a good grasp of what was actually practicable. The boring but necessary daily work on the home front was not very interesting to him, so he willingly handed it over to others. He hardly noticed the lack of administrative machinery, a prerequisite for any scholarly museum. It was not easy to follow him on his flights of fancy and still build up the necessary support base. However, the fact that he in no way spared himself had an appeasing effect.Provincial museums at that time were of a mixed nature. A few had trained management, and the rest were run by interested locals. This was often excellently done, as in Esbjerg, where the master joiner Niels Thomsen and a staff of volunteers carried out excavations that were as good as professional investigations, and published them in well-written articles. Regrettably, there were also examples of the opposite. A museum curator in Jutland informed me that his predecessor had been an eager excavator but very rarely left any written documentation of his actions. The excavated items were left without labels in the museum store, often wrapped in newspapers. However, these gave a clue as to the time of unearthing, and with a bit of luck a look in the newspaper archive would then reveal where the excavation had taken place. Although somewhat exceptional, this is not the only such case.The Museum of Aarhus definitely belonged among the better ones in this respect. Founded in 1861, it was at first located at the then town hall, together with the local art collection. The rooms here soon became too cramped, and both collections were moved to a new building in the ”Mølleparken” park. There were skilful people here working as managers and assistants, such as Vilhelm Boye, who had received his archaeological training at the National Museum, and later the partners A. Reeh, a barrister, and G.V. Smith, a captain, who shared the honour of a number of skilfully performed excavations. Glob’s predecessor as curator was the librarian Ejler Haugsted, also a competent man of fine achievements. We did not, thus, take over a museum on its last legs. On the other hand, it did not meet the requirements of a modern scholarly museum. We were given the task of turning it into such a museum, as implied by the name change.The goal was to create a museum similar to the National Museum, but without the faults and shortcomings that that museum had developed over a period of time. In this respect our nightly conversations during our years in Copenhagen turned out to be useful, as our talk had focused on these imperfections and how to eradicate them.We now had the opportunity to put our theories into practice. We may not have succeeded in doing so, but two areas were essentially improved:The numerous independent numbering systems, which were familiar to us from the National Museum, were permeating archaeological excavation s not only in the field but also during later work at the museum. As far as possible this was boiled down to a single system, and a new type of report was born. (In this context, a ”report” is the paper following a field investigation, comprising drawings, photos etc. and describing the progress of the work and the observations made.) The instructions then followed by the National Museum staff regarding the conduct of excavations and report writing went back to a 19th-century protocol by the employee G.V. Blom. Although clear and rational – and a vast improvement at the time – this had become outdated. For instance, the excavation of a burial mound now involved not only the middle of the mound, containing the central grave and its surrounding artefacts, but the complete structure. A large number of details that no one had previously paid attention to thus had to be included in the report. It had become a comprehensive and time-consuming work to sum up the desultory notebook records in a clear and understandable description.The instructions resulting from the new approach determined a special records system that made it possible to transcribe the notebook almost directly into a report following the excavation. The transcription thus contained all the relevant information concerning the in vestigation, and included both relics and soil layers, the excavation method and practical matters, although in a random order. The report proper could then bereduced to a short account containing references to the numbers in the transcribed notebook, which gave more detailed information.As can be imagined, the work of reform was not a continuous process. On the contrary, it had to be done in our spare hours, which were few and far between with an employer like Glob. The assignments crowded in, and the large Jutland map that we had purchased was as studded with pins as a hedge hog’s spines. Each pin represented an inuninent survey, and many of these grew into small or large excavations. Glob himself had his lecture duties to perform, and although he by no means exaggerated his concern for the students, he rarely made it further than to the surveys. Bibby and I had to deal with the hard fieldwork. And the society, once it was established, did not make our lives any easier. Kuml demanded articles written at lightning speed. A perusal of my then diary has given me a vivid recollection of this hectic period, in which I had to make use of the evening and night hours, when the museum was quiet and I had a chance to collect my thoughts. Sometimes our faithful supporter, the Lord Mayor, popped in after an evening meeting. He was extremely interested in our problems, which were then solved according to our abilities over a cup of instant coffee.A large archaeological association already existed in Denmark. How ever, Glob found it necessary to establish another one which would be less oppressed by tradition. Det kongelige nordiske Oldsskriftselskab had been funded in 1825 and was still influenced by different peculiarities from back then. Membership was not open to everyone, as applications were subject to recommendation from two existing members and approval by a vote at one of the monthly lecture meetings. Most candidates were of course accepted, but unpopular persons were sometimes rejected. In addition, only men were admitted – women were banned – but after the war a proposal was brought forward to change this absurdity. It was rejected at first, so there was a considerable excitement at the January meeting in 1951, when the proposal was once again placed on the agenda. The poor lecturer (myself) did his best, although he was aware of the fact that just this once it was the present and not the past which was the focus of attention. The result of the voting was not very courteous as there were still many opponents, but the ladies were allowed in, even if they didn’t get the warmest welcome.In Glob’s society there were no such restrictions – everyone was welcome regardless of sex or age. If there was a model for the society, it was the younger and more progressive Norwegian Archaeological Society rather than the Danish one. The main purpose of both societies was to produce an annual publication, and from the start Glob’s Kuml had a closer resemblance to the Norwegian Viking than to the Danish Aarbøger for nordisk Oldkyndighed og Historie. The name of the publication caused careful consideration. For a long time I kept a slip of paper with different proposals, one of which was Kuml, which won after having been approved by the linguist Peter Skautrup.The name alone, however, was not enough, so now the task became to find so mething to fill Kuml with. To this end the finds came in handy, and as for those, Glob must have allied him self with the higher powers, since fortune smiled at him to a considerable extent. Just after entering upon his duties in Aarhus, an archaeological sensation landed at his feet. This happened in May 1950 when I was still living in the capital. A few of us had planned a trip to Aarhus, partly to look at the relics of th e past, and partly to visit our friend, the professor. He greeted us warmly and told us the exciting news that ten iron swords had been found during drainage work in the valley of lllerup Aadal north of the nearby town of Skanderborg. We took the news calmly as Glob rarely understated his affairs, but our scepticism was misplaced. When we visited the meadow the following day and carefully examined the dug-up soil, another sword appeared, as well as several spear and lance heads, and other iron artefacts. What the drainage trench diggers had found was nothing less than a place of sacrifice for war booty, like the four large finds from the 1800s. When I took up my post in Aarhus in September of that year I was granted responsibility for the lllerup excavation, which I worked on during the autumn and the following six summers. Some of my best memories are associated with this job – an interesting and happy time, with cheerful comradeship with a mixed bunch of helpers, who were mainly archaeology students. When we finished in 1956, it was not because the site had been fully investigated, but because the new owner of the bog plot had an aversion to archaeologists and their activities. Nineteen years later, in 1975, the work was resumed, this time under the leadership of Jørgen Ilkjær, and a large amount of weaponry was uncovered. The report from the find is presently being published.At short intervals, the year 1952 brought two finds of great importance: in Februar y the huge vessel from Braa near Horsens, and in April the Grauballe Man. The large Celtic bronze bowl with the bulls’ heads was found disassembled, buried in a hill and covered by a couple of large stones. Thanks to the finder, the farmer Søren Paaske, work was stopped early enough to leave areas untouched for the subsequent examination.The saga of the Grauballe Man, or the part of it that we know, began as a rumour on the 26th of April: a skeleton had been found in a bog near Silkeborg. On the following day, which happened to be a Sunday, Glob went off to have a look at the find. I had other business, but I arrived at the museum in the evening with an acquaintance. In my diary I wrote: ”When we came in we had a slight shock. On the floor was a peat block with a corpse – a proper, well-preserved bog body. Glob brought it. ”We’ll be in trouble now.” And so we were, and Glob was in high spirits. The find created a sensation, which was also thanks to the quick presentation that we mounted. I had purchased a tape recorder, which cost me a packet – not a small handy one like the ones you get nowadays, but a large monstrosity with a steel tape (it was, after all, early days for this device) – and assisted by several experts, we taped a number of short lectures for the benefit of the visitors. People flocked in; the queue meandered from the exhibition room, through the museum halls, and a long way down the street. It took a long wait to get there, but the visitors seemed to enjoy the experience. The bog man lay in his hastily – procured exhibition case, which people circled around while the talking machine repeatedly expressed its words of wisdom – unfortunately with quite a few interruptions as the tape broke and had to be assembled by hand. Luckily, the tape recorders now often used for exhibitions are more dependable than mine.When the waves had died down and the exhibition ended, the experts examined the bog man. He was x-rayed at several points, cut open, given a tooth inspection, even had his fingerprints taken. During the autopsy there was a small mishap, which we kept to ourselves. However, after almost fifty years I must be able to reveal it: Among the organs removed for investigation was the liver, which was supposedly suitable for a C-14 dating – which at the time was a new dating method, introduced to Denmark after the war. The liver was sent to the laboratory in Copenhagen, and from here we received a telephone call a few days later. What had been sent in for examination was not the liver, but the stomach. The unfortunate (and in all other respects highly competent) Aarhus doctor who had performed the dissection was cal1ed in again. During another visit to the bogman’s inner parts he brought out what he believed to be the real liver. None of us were capable of deciding th is question. It was sent to Copenhagen at great speed, and a while later the dating arrived: Roman Iron Age. This result was later revised as the dating method was improved. The Grauballe Man is now thought to have lived before the birth of Christ.The preservation of the Grauballe Man was to be conservation officer Kornbak’s masterpiece. There were no earlier cases available for reference, so he invented a new method, which was very successful. In the first volumes of Kuml, society members read about the exiting history of the bog body and of the glimpses of prehistoric sacrificial customs that this find gave. They also read about the Bahrain expeditions, which Glob initiated and which became the apple of his eye. Bibby played a central role in this, as it was he who – at an evening gathering at Glob’s and Harriet’s home in Risskov – described his stay on the Persian Gulf island and the numerous burial mounds there. Glob made a quick decision (one of his special abilities was to see possibilities that noone else did, and to carry them out successfully to everyone’s surprise) and in December 1952 he and Bibby left for the Gulf, unaware of the fact that they were thereby beginning a series of expeditions which would continue for decades. Again it was Glob’s special genius that was the decisive factor. He very quickly got on friendly terms with the rulers of the small sheikhdoms and interested them in their past. As everyone knows, oil is flowing plentifully in those parts. The rulers were thus financially powerful and some of this wealth was quickly diverted to the expeditions, which probably would not have survived for so long without this assistance. To those of us who took part in them from time to time, the Gulf expeditions were an unforgettable experience, not just because of the interesting work, but even more because of the contact with the local population, which gave us an insight into local manners and customs that helped to explain parts of our own country’s past which might otherwise be difficult to understand. For Glob and the rest of us did not just get close to the elite: in spite of language problems, our Arab workers became our good friends. Things livened up when we occasionally turned up in their palm huts.Still, co-operating with Glob was not always an easy task – the sparks sometimes flew. His talent of initiating things is of course undisputed, as are the lasting results. He was, however, most attractive when he was in luck. Attention normally focused on this magnificent person whose anecdotes were not taken too seriously, but if something went wrong or failed to work out, he could be grossly unreasonable and a little too willing to abdicate responsibility, even when it was in fact his. This might lead to violent arguments, but peace was always restored. In 1954, another museum curator was attached to the museum: Poul Kjærum, who was immediately given the important task of investigating the dolmen settlement near Tustrup on Northern Djursland. This gave important results, such as the discovery of a cult house, which was a new and hitherto unknown Stone Age feature.A task which had long been on our mind s was finally carried out in 1955: constructing a new display of the museum collections. The old exhibitio n type consisted of numerous artefacts lined up in cases, accompaied ony by a brief note of the place where it was found and the type – which was the standard then. This type of exhibition did not give much idea of life in prehistoric times.We wanted to allow the finds to speak for themselves via the way that they were arranged, and with the aid of models, photos and drawings. We couldn’t do without texts, but these could be short, as people would understand more by just looking at the exhibits. Glob was in the Gulf at the time, so Kjærum and I performed the task with little money but with competent practical help from conservator Kornbak. We shared the work, but in fairness I must add that my part, which included the new lllerup find, was more suitable for an untraditional display. In order to illustrate the confusion of the sacrificial site, the numerous bent swords and other weapons were scattered a.long the back wall of the exhibition hall, above a bog land scape painted by Emil Gregersen. A peat column with inlaid slides illustrated the gradual change from prehistoric lake to bog, while a free-standing exhibition case held a horse’s skeleton with a broken skull, accompanied by sacrificial offerings. A model of the Nydam boat with all its oars sticking out hung from the ceiling, as did the fine copy of the Gundestrup vessel, as the Braa vessel had not yet been preserved. The rich pictorial decoration of the vessel’s inner plates was exhibited in its own case underneath. This was an exhibition form that differed considerably from all other Danish exhibitions of the time, and it quickly set a fashion. We awaited Glob’s homecoming with anticipation – if it wasn’t his exhibition it was still made in his spirit. We hoped that he would be surprised – and he was.The museum was thus taking shape. Its few employees included Jytte Ræbild, who held a key position as a secretary, and a growing number of archaeology students who took part in the work in various ways during these first years. Later, the number of employees grew to include the aforementioned excavation pioneer Georg Kunwald, and Hellmuth Andersen and Hans Jørgen Madsen, whose research into the past of Aarhus, and later into Danevirke is known to many, and also the ethnographer Klaus Ferdinand. And now Moesgaard appeared on the horizon. It was of course Glob’s idea to move everything to a manor near Aarhus – he had been fantasising about this from his first Aarhus days, and no one had raised any objections. Now there was a chance of fulfilling the dream, although the actual realisation was still a difficult task.During all this, the Jutland Archaeological Society thrived and attracted more members than expected. Local branches were founded in several towns, summer trips were arranged and a ”Worsaae Medal” was occasionally donated to persons who had deserved it from an archaeological perspective. Kuml came out regularly with contributions from museum people and the like-minded. The publication had a form that appealed to an inner circle of people interested in archaeology. This was the intention, and this is how it should be. But in my opinion this was not quite enough. We also needed a publication that would cater to a wider public and that followed the same basic ideas as the new exhibition.I imagined a booklet, which – without over-popularsing – would address not only the professional and amateur archaeologist but also anyone else interested in the past. The result was Skalk, which (being a branch of the society) published its fir t issue in the spring of 1957. It was a somewhat daring venture, as the financial base was weak and I had no knowledge of how to run a magazine. However, both finances and experience grew with the number of subscribers – and faster than expected, too. Skalk must have met an unsatisfied need, and this we exploited to the best of our ability with various cheap advertisements. The original idea was to deal only with prehistoric and medieval archaeology, but the historians also wanted to contribute, and not just the digging kind. They were given permission, and so the topic of the magazine ended up being Denmark’s past from the time of its first inhabitant s until the times remembered by the oldest of us – with the odd sideways leap to other subjects. It would be impossible to claim that Skalk was at the top of Glob’s wish list, but he liked it and supported the idea in every way. The keeper of national antiquities, Johannes Brøndsted, did the same, and no doubt his unreserved approval of the magazine contributed to its quick growth. Not all authors found it easy to give up technical language and express themselves in everyday Danish, but the new style was quickly accepted. Ofcourse the obligations of the magazine work were also sometimes annoying. One example from the diary: ”S. had promised to write an article, but it was overdue. We agreed to a final deadline and when that was overdue I phoned again and was told that the author had gone to Switzerland. My hair turned grey overnight.” These things happened, but in this particular case there was a happy ending. Another academic promised me three pages about an excavation, but delivered ten. As it happened, I only shortened his production by a third.The 1960s brought great changes. After careful consideration, Glob left us to become the keeper of national antiquities. One important reason for his hesitation was of course Moesgaard, which he missed out on – the transfer was almost settled. This was a great loss to the Aarhus museum and perhaps to Glob, too, as life granted him much greater opportunities for development.” I am not the type to regret things,” he later stated, and hopefully this was true. And I had to choose between the museum and Skalk – the work with the magazine had become too timeconsuming for the two jobs to be combined. Skalk won, and I can truthfully say that I have never looked back. The magazine grew quickly, and happy years followed. My resignation from the museum also meant that Skalk was disengaged from the Jutland Archaeological Society, but a close connection remained with both the museum and the society.What has been described here all happened when the museum world was at the parting of the ways. It was a time of innovation, and it is my opinion that we at the Prehistoric Museum contributed to that change in various ways.The new Museum Act of 1958 gave impetus to the study of the past. The number of archaeology students in creased tremendously, and new techniques brought new possibilities that the discussion club of the 1940s had not even dreamt of, but which have helped to make some of the visions from back then come true. Public in terest in archaeology and history is still avid, although to my regret, the ahistorical 1960s and 1970s did put a damper on it.Glob is greatly missed; not many of his kind are born nowadays. He had, so to say, great virtues and great fault s, but could we have done without either? It is due to him that we have the Jutland Archaeological Society, which has no w existed for half a century. Congr tulat ion s to the Society, from your offspring Skalk.Harald AndersenSkalk MagazineTranslated by Annette Lerche Trolle

Abstract. Sulaksono N, Pudyatmoko S, Soemardi, Wardhana W, Hadiyan Y, Nurvianto S. 2022. Response of terrestrial mammals to various types of disturbance in the Gunung Merapi National Park, Indonesia. Biodiversitas 23: 1635-1647. Human disturbance in the form of sand mining and grass harvesting activities in the area of Mount Merapi is an important component of the ecological process alongside the eruption. The group of terrestrial mammals (TM) is one of the essential elements that are affected by this disturbance. Therefore, the objectives of this study were to determine the response of the TM in various types of habitats in areas affected by eruption disturbances >20 years (ERL), eruption < 20 years (ERE), mining (MIN), grazing (GRA), and non-disturbance areas (NDA) in Gunung Merapi National Park (GMNP). Camera traps were used to identify the abundance of each TM. Vegetation data were also used to represent habitat conditions and were taken on a plot of 0.04 Ha. The analysis results showed that the highest abundance of TM is at the NDA site (14.4), while the lowest is at the MIN site (2.08). The site with the highest level of TM diversity is ERL (1.97) and the lowest is NDA (1.52). Furthermore, there is a significant difference in TM abundance between the MIN and ERL sites, which indicates that each disturbance reacts differently to the site. Based on the results, mining disturbances negatively influenced the abundance of TM in GMNP, therefore, managers need to regulate mining activities and make efforts to restore the affected areas.

Abstract. Wulandari C, Novriyanti N, Iswandaru D. 2021. Integrating ecological, social and policy aspects to develop peatland restoration strategies in Orang Kayo Hitam Forest Park, Jambi, Indonesia. Biodiversitas 22: 4158-4168. Peatlands play an essential role in delivering various ecosystem services, yet in many areas in Indonesia, peat forests have been deforested, degraded, and threatened by repeated fires. As such, restoration efforts need to be done. But these require various considerations, including ecological aspects such as type degradation conditions, water table, species selected for planting, and climates, socio-economic aspects like community's involvement, and policy aspects such as regulations and institutions at national and local levels. This study aimed to analyze peatland restoration strategies in Orang Kayo Hitam (OKH) Forest Park, Jambi using the Strengths, Weaknesses, Threats, and Opportunities (SWOT) framework. The analysis was done by integrating multiple aspects such as policy, biodiversity conservation, and surrounding community participation. Data were collected using virtual ethnographic techniques due to the COVID-19 pandemic. Key informants (i.e., OKH Forest Park Unit area managers, NGOs, staff of national government, and university academics) and selected respondents were interviewed to identify and determine factors considered as internal (i.e., Strengths and Weaknesses) and external (i.e., Threats and Opportunities). This qualitative data was then analyzed using a SWOT approach through the IFE (Internal Factor Evaluation) and EFE (External Factor Evaluation) Matrix. The research findings showed that from 12 internal factors and 11 external factors, efforts to restore peatland ecosystems in Orang Kayo Hitam (OKH) Forest Park had obtained very strong values in the biodiversity aspect. The SWOT matrix analysis showed values of S-W = 0.43 and O-T = 0.16 positioned in quadrant I. This result suggests that the most powerful strategies can be developed aggressively from the factors of strengths and opportunities.

Abstract. Wulandari C, Novriyanti N, Iswandaru D. 2021. Integrating ecological, social and policy aspects to develop peatland restoration strategies in Orang Kayo Hitam Forest Park, Jambi, Indonesia. Biodiversitas 22: 4158-4168. Peatlands play an essential role in delivering various ecosystem services, yet in many areas in Indonesia, peat forests have been deforested, degraded, and threatened by repeated fires. As such, restoration efforts need to be done. But these require various considerations, including ecological aspects such as type degradation conditions, water table, species selected for planting, and climates, socio-economic aspects like community's involvement, and policy aspects such as regulations and institutions at national and local levels. This study aimed to analyze peatland restoration strategies in Orang Kayo Hitam (OKH) Forest Park, Jambi using the Strengths, Weaknesses, Threats, and Opportunities (SWOT) framework. The analysis was done by integrating multiple aspects such as policy, biodiversity conservation, and surrounding community participation. Data were collected using virtual ethnographic techniques due to the COVID-19 pandemic. Key informants (i.e., OKH Forest Park Unit area managers, NGOs, staff of national government, and university academics) and selected respondents were interviewed to identify and determine factors considered as internal (i.e., Strengths and Weaknesses) and external (i.e., Threats and Opportunities). This qualitative data was then analyzed using a SWOT approach through the IFE (Internal Factor Evaluation) and EFE (External Factor Evaluation) Matrix. The research findings showed that from 12 internal factors and 11 external factors, efforts to restore peatland ecosystems in Orang Kayo Hitam (OKH) Forest Park had obtained very strong values in the biodiversity aspect. The SWOT matrix analysis showed values of S-W = 0.43 and O-T = 0.16 positioned in quadrant I. This result suggests that the most powerful strategies can be developed aggressively from the factors of strengths and opportunities.

Abstract. Pla-ard M, Khioesree N, Sungkalak B, Nathalang A, Thomas W, Uthairatsamee S, Paansri P, Chanachai Y, Sukmasuang R. 2021. Population characteristics and habitat suitability of Khao Yai National Park, Thailand for Asian elephant and five ungulate species. Biodiversitas 23: 231-243. This study of population characteristics and habitat suitability of Asian elephant (Elephas maximus Linnaeus, 1758) and five ungulate species was conducted between October 2017 and July 2020 in Khao Yai National Park (KYNP) using camera traps. One hundred and twenty-two camera trap locations were set up for a total of 4,139 trap nights and 5,461 independent encounters were identified. The target species of Asian elephant, gaur, sambar deer, wild boar, northern red muntjac and lesser oriental chevrotain were recorded. The results show the occupancy of Asian elephant was 1.21 individuals/km2 within the study area. The probability occupancy of the Asian elephant was 70% (SE=0.06). In comparison, gaur had a probability occupancy of 57% (SE=0.07), whereas sambar deer was 79% (SE=0.04), followed by wild boar 77% (SE=0.05), northern red muntjac 77% (SE=0.05) and lesser oriental chevrotain occupancy 63% (SE=0.07). The age structure of Asian elephant between calf, juvenile, sub-adult and adult was 1: 2.1: 1.2: 3.16, and the ratio between adult males and females was 1: 1.72. The results show that roads and salt lick sites were the essential factors affecting the chance of Asian elephants and ungulate species in the area. The habitat suitability for Asian elephant was 331 km2, while those for gaur, sambar deer, wild boar, northern red muntjac and lesser oriental chevrotain were 287.73 km2, 249.97 km2, 540.40 km2, 451.34 km2 and 434.30 km2, respectively. Recommendations for further management involve concentration on the suitable area resulting from this study. Concerning the suitability habitat, it was found that the park boundary was most suited. Therefore, habitat improvements for all large herbivorous mammal species should improve the areas within the national park and especially address the central area, with an emphasis on creating salt lick sites, in addition to grassland and water sources that must be quality and sufficient, the most important habitat factor for these species.

Today, the incidence of type 2 diabetes mellitus is increasing rapidly on global. This disease is shown with many complications that significantly affect public health. One of them is kidney complications, which have a high incidence among diabetic patients in Vietnam (25.6-33.1%). Age, history of hypertension, and dyslipidemia are considered to be the main risk factors for diabetic nephropathy. Thus, early detection of these factors for kidney damage is significant for diagnosing, monitoring, treatment, and prognosis of diabetic patients. Our descriptive, cross-sectional study conducting on 120 diabetic patients at E Hospital has observed that blood cholesterol levels, HbA1c levels were independently related to eGFR decline below 60 mL/min/1.73m2. From those data, an equation to predict the risk of diabetic kidney disease was estimated as p = with k = Keyword: Type 2 diabetes, Diabetic nephropathy, Risk factor Today, the incidence of type 2 diabetes mellitus is increasing rapidly on global. This disease is shown with many complications that significantly affect public health. One of them is kidney complications, which have a high incidence among diabetic patients in Vietnam (25.6-33.1%). Age, history of hypertension, and dyslipidemia are considered to be the main risk factors for diabetic nephropathy. Thus, early detection of these factors for kidney damage is significant for diagnosing, monitoring, treatment, and prognosis of diabetic patients. Our descriptive, cross-sectional study conducting on 120 diabetic patients at E Hospital has observed that blood cholesterol levels, HbA1c levels were independently related to eGFR decline below 60 mL/min/1.73m2. 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Previous studies contend that Philippines is still a ‘collectivist’ society (Cf. Hofstede Center; Cukur et al. 2004:613-634). In this collectivist or community-oriented society, individualism is not something that is highly valued. Being ‘individualistic’ is often associated to being narcissistic, loner, asocial, selfish, etc. However, one may ask whether the youth in the Philippines are not spared from this insidious culture of individualism, notwithstanding the seemingly dominant collective and communitarian character of the society. Although the overwhelming poverty is still the main problem in the Philippines, where according to Wostyn (2010:26) “only the wonderland of movies gives some respite to their consciousness of suffering and oppression”, the Filipino youth of today are also exposed to the consumeristic values of the ‘city’ and are not spared from the contradictions and insecurities posed by the pluralistic society. They are citizens of an increasing social and cultural pluralism characteristic of many liberal societies. Is it possible that individualism may also exist within this culture, especially among the younger generation? Is individualism slowly creeping in as caused by their exposure and easy access to modern technology, to higher education, mobility, interactions with other cultures, etc. Would this individualistic tendency have any influence on their religious beliefs, especially their belief on salvation? What would be the implications and challenges of these findings to the families in the Philippines? These are the questions we wish to answer in this study. This paper is structured in four parts: first, we will discuss the theoretical framework of individualism and salvation; second, we will examine the empirical attitudes on individualism and salvation; third, we will explore the relationship between individualism and salvation; and finally, we will draw some pastoral implication especially in relation to the document “Lineamenta - The Vocation and Mission of the Family in the church and Contemporary Word” (henceforth, Lineamenta). References Atkins, P. (2004). Memory and Liturgy. The Place of Memory in the Composition and Practice of Liturgy. Hampshire: Ashgate Publishing. Bauman, Z. (1993). Postmodern Ethics. Oxford/Cambridge, MA: Blackwell. Beck, U. (1992). Risk society. London: Sage Publications. Bellah, R. N. , Madsen, R., Sullivan, W., Swidler, A., Tipton, S. (1985). Habits of the Heart: Individualism and Commitment in American Life. Berkeley/Los Angeles/London: University of California Press. Berger, P. (1970). 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This article reflects on part of a three-year battle over the redevelopment of an iconic Melbourne music venue, the Palace-Metro Nightclub (the Palace), involving the tactical use of Facebook Page data at trial. We were invited by the Save the Palace group, Melbourne City Council and the National Trust of Australia to provide Facebook Page data analysis as evidence of the social value of the venue at an appeals trial heard at the Victorian Civil Administration Tribunal (VCAT) in 2016. We take a reflexive ethnographic approach here to explore the data production, collection and analysis processes as these represent and constitute a “data public”.Although the developers won the appeal and were able to re-develop the site, the court accepted the validity of social media data as evidence of the building’s social value (Jinshan Investment Group Pty Ltd v Melbourne CC [2016] VCAT 626, 117; see also Victorian Planning Reports). Through the case, we elaborate on the concept of data publics by considering the “affordising” (Pollock) processes at play when extracting, analysing and visualising social media data. Affordising refers to the designed, deliberate and incidental effects of datafication and highlights the need to attend to the capacities for data collection and processing as they produce particular analytical outcomes. These processes foreground the compositional character of data publics, and the unevenness of data literacies (McCosker “Data Literacies”; Gray et al.) as a factor of the interpersonal and institutional capacity to read and mobilise data for social outcomes.We begin by reconsidering the often-assumed connection between social media data and their publics. Taking onboard theoretical accounts of publics as problem-oriented (Dewey) and dynamically constituted (Kelty), we conceptualise data publics through the key elements of a) consequentiality, b) sufficient connection over time, c) affective or emotional qualities of connection and interaction with the events. We note that while social data analytics may be a powerful tool for public protest, it equally affords use against public interests and introduces risks in relation to a lack of transparency, access or adequate data literacy.Urban Protest and Data Publics There are many examples globally of the use of social media to engage publics in battles over urban development or similar issues (e.g. Fredericks and Foth). Some have asked how social media might be better used by neighborhood organisations to mobilise protest and save historic buildings, cultural landmarks or urban sites (Johnson and Halegoua). And we can only note here the wealth of research literature on social movements, protest and social media. To emphasise Gerbaudo’s point, drawing on Mattoni, we “need to account for how exactly the use of these media reshapes the ‘repertoire of communication’ of contemporary movements and affects the experience of participants” (2). For us, this also means better understanding the role that social data plays in both aiding and reshaping urban protest or arming third sector groups with evidence useful in social institutions such as the courts.New modes of digital engagement enable forms of distributed digital citizenship, which Meikle sees as the creative political relationships that form through exercising rights and responsibilities. Associated with these practices is the transition from sanctioned, simple discursive forms of social protest in petitions, to new indicators of social engagement in more nuanced social media data and the more interactive forms of online petition platforms like change.org or GetUp (Halpin et al.). These technical forms code publics in specific ways that have implications for contemporary protest action. That is, they provide the operational systems and instructions that shape social actions and relationships for protest purposes (McCosker and Milne).All protest and social movements are underwritten by explicit or implicit concepts of participatory publics as these are shaped, enhanced, or threatened by communication technologies. But participatory protest publics are uneven, and as Kelty asks: “What about all the people who are neither protesters nor Twitter users? In the broadest possible sense this ‘General Public’ cannot be said to exist as an actual entity, but only as a kind of virtual entity” (27). Kelty is pointing to the porous boundary between a general public and an organised public, or formal enterprise, as a reminder that we cannot take for granted representations of a public, or the public as a given, in relation to Like or follower data for instance.If carefully gauged, the concept of data publics can be useful. To start with, the notions of publics and publicness are notoriously slippery. Baym and boyd explore the differences between these two terms, and the way social media reconfigures what “public” is. Does a Comment or a Like on a Facebook Page connect an individual sufficiently to an issues-public? As far back as the 1930s, John Dewey was seeking a pragmatic approach to similar questions regarding human association and the pluralistic space of “the public”. For Dewey, “the machine age has so enormously expanded, multiplied, intensified and complicated the scope of the indirect consequences [of human association] that the resultant public cannot identify itself” (157). To what extent, then, can we use data to constitute a public in relation to social protest in the age of data analytics?There are numerous well formulated approaches to studying publics in relation to social media and social networks. Social network analysis (SNA) determines publics, or communities, through links, ties and clustering, by measuring and mapping those connections and to an extent assuming that they constitute some form of sociality. Networked publics (Ito, 6) are understood as an outcome of social media platforms and practices in the use of new digital media authoring and distribution tools or platforms and the particular actions, relationships or modes of communication they afford, to use James Gibson’s sense of that term. “Publics can be reactors, (re)makers and (re)distributors, engaging in shared culture and knowledge through discourse and social exchange as well as through acts of media reception” (Ito 6). Hashtags, for example, facilitate connectivity and visibility and aid in the formation and “coordination of ad hoc issue publics” (Bruns and Burgess 3). Gray et al., following Ruppert, argue that “data publics are constituted by dynamic, heterogeneous arrangements of actors mobilised around data infrastructures, sometimes figuring as part of them, sometimes emerging as their effect”. The individuals of data publics are neither subjugated by the logics and metrics of digital platforms and data structures, nor simply sovereign agents empowered by the expressive potential of aggregated data (Gray et al.).Data publics are more than just aggregates of individual data points or connections. They are inherently unstable, dynamic (despite static analysis and visualisations), or vibrant, and ephemeral. We emphasise three key elements of active data publics. First, to be more than an aggregate of individual items, a data public needs to be consequential (in Dewey’s sense of issues or problem-oriented). Second, sufficient connection is visible over time. Third, affective or emotional activity is apparent in relation to events that lend coherence to the public and its prevailing sentiment. To these, we add critical attention to the affordising processes – or the deliberate and incidental effects of datafication and analysis, in the capacities for data collection and processing in order to produce particular analytical outcomes, and the data literacies these require. We return to the latter after elaborating on the Save the Palace case.Visualising Publics: Highlighting Engagement and IntensityThe Palace theatre was built in 1912 and served as a venue for theatre, cinema, live performance, musical acts and as a nightclub. In 2014 the Heritage Council decided not to include the Palace on Victoria’s heritage register and hence opened the door for developers, but Melbourne City Council and the National Trust of Australia opposed the redevelopment on the grounds of the building’s social significance as a music venue. Similarly, the Save the Palace group saw the proposed redevelopment as affecting the capacity of Melbourne CBD to host medium size live performances, and therefore impacting deeply on the social fabric of the local music scene. The Save the Palace group, chaired by Rebecca Leslie and Michael Raymond, maintained a 36,000+ strong Facebook Page and mobilised local members through regular public street protests, and participated in court proceedings in 2015 and February 2016 with Melbourne City Council and National Trust Australia. Joining the protesters in the lead up to the 2016 appeals trial, we aimed to use social media engagement data to measure, analyse and present evidence of the extent and intensity of a sustained protest public. The evidence we submitted had to satisfy VCAT’s need to establish the social value of the building and the significance of its redevelopment, and to explain: a) how social media works; b) the meaning of the number of Facebook Likes on the Save The Palace Page and the timing of those Likes, highlighting how the reach and Likes pick up at significant events; and c) whether or not a representative sample of Comments are supportive of the group and the Palace Theatre (McCosker “Statement”). As noted in the case (Jinshan, 117), where courts have traditionally relied on one simple measure for contemporary social value – the petition – our aim was to make use of the richer measures available through social media data, to better represent sustained engagement with the issues over time.Visualising a protest public in this way raises two significant problems for a workable concept of data publics. The first involves the “affordising” (Pollock) work of both the platform and our data analysis. This concerns the role played by data access and platform affordances for data capture, along with methodological choices made to best realise or draw out the affordances of the data for our purposes. The second concerns the issue of digital and data literacies in both the social acts that help to constitute a data public in the first place, and the capacity to read and write public data to represent those activities meaningfully. That is, Facebook and our analysis constitutes a data public in certain ways that includes potentially opaque decisions or processes. And citizens (protesters or casual Facebook commenters alike) along with social institutions (like the courts) have certain uneven capacity to effectively produce or read public protest-oriented data. The risk here, which we return to in the final section, lies in the potential for misrepresentation of publics through data, exclusions of access and ownership of data, and the uneven digital literacies at each stage of data production, analysis and sensemaking.Facebook captures data about individuals in intricate detail. Its data capture strategies are geared toward targeting for the purposes of marketing, although only a small subset of the data is publicly available through the Facebook Application Programming Interface (API), which is a kind of data “gateway”. The visible page data tells only part of the story. The total Page Likes in February 2016 was 36,828, representing a sizeable number of followers, mainly located in Melbourne but including 45 countries in total and 38 different languages. We extracted a data set of 268,211 engagements with the Page between February 2013 and August 2015. This included 45,393 post Likes and 9,139 Comments. Our strategy was to demarcate a structurally defined “community” (in the SNA sense of that term as delineating clusters of people, activities and links within a broader network), by visualising the interactions of Facebook users with Posts over time, and then examine elements of intensity of engagement. In other words, we “affordised” the network data using SNA techniques to most clearly convey the social value of the networked public.We used a combination of API access and Facebook’s native Insights data and analytics to extract use-data from that Page between June 2013 and December 2015. Analysis of a two-mode or bipartite network consisting of users and Posts was compiled using vosonSML, a package in the R programming language created at Australian National University (Graham and Ackland) and visualised with Gephi software. In this network, the nodes (or vertices) represent Facebook users and Facebook Posts submitted on the Page, and ties (or edges) between nodes represent whether a user has commented on and/or liked a post. For example, a user U might have liked Post A and commented on Post B. Additionally, a weight value is assigned for the Comments ties, indicating how many times a user commented on a particular post (note that users can only like Posts once). We took these actions as demonstrating sufficient connection over time in relation to an issue of common concern.Figure 1: Network visualisation of activity on the Save the Palace Facebook Page, June 2013 to December 2015. The colour of the nodes denotes which ‘community’ cluster they belong to (computed via the Infomap algorithm) and nodes are sized by out-degree (number of Likes/Comments made by users to Posts). The graph layout is computed via the Force Atlas 2 algorithm.Community detection was performed on the network using the Infomap algorithm (Rosvall and Bergstrom), which is suited to large-scale weighted and directed networks (Henman et al.). This analysis reveals two large and two smaller clusters or groups represented by colour differences (Fig. 1). Broadly, this suggests the presence of several clusters amongst a sustained network engaging with the page over the three years. Beyond this, a range of other colours denoting smaller clusters indicates a diversity of activity and actors co-participating in the network as part of a broader community.The positioning of nodes within the network is not random – the visualisation is generated by the Force Atlas 2 algorithm (Jacomy et al.) that spatially sorts the nodes through processes of attraction and repulsion according to the observed patterns of connectivity. As we would expect, the two-dimensional spatial arrangement of nodes conforms to the community clustering, helping us to visualise the network in the form of a networked public, and build a narrative interpretation of “what is going on” in this online social space.Social value for VCAT was loosely defined as a sense of connection, sentiment and attachment to the venue. While we could illustrate the extent of the active connections of those engaging with the Page, the network map does not in itself reveal much about the sentiment, or the emotional attachment to the Save the Palace cause. This kind of affect can be understood as “the energy that drives, neutralizes, or entraps networked publics” (Papacharissi 7), and its measure presents a particular challenge, but also interest, for understanding a data public. It is often measured through sentiment analysis of content, but we targeted reach and engagement events – particular moments that indicated intense interaction with the Page and associated events.Figure 2: Save the Palace Facebook Page: Organic post reach November—December 2014The affective connection and orientation could be demonstrated through two dimensions of post “reach”: average reach across the lifespan of the Page, and specific “reach-events”. Average reach illustrates the sustained engagement with the Page over time. Average un-paid reach for Posts with links (primarily news and legal updates), was 12,015 or 33% of the total follower base – a figure well above the standard for Community Page reach at that time. Reach-events indicated particular points of intensity and illustrates the Page’s ability to resonate publicly. Figure 2 points to one such event in November 2015, when news circulated that the developers were defying stop-work orders and demolishing parts of The Palace. The 100k reach indicated intense and widespread activity – Likes, Shares, Comments – in a short timeframe. We examined Comment activity in relation to specific reach events to qualify this reach event and illustrate the sense of outrage directed toward the developers, and expressions of solidarity toward those attempting to stop the redevelopment. Affordising Data Publics and the Transformative Work of AnalyticsEach stage of deriving evidence of social value through Page data, from building public visibility and online activity to analysis and presentation at VCAT, was affected by the affordising work of the protesters involved (particularly the Page Admins), civil society groups, platform features and data structures and our choices in analysis and presentation. The notion of affordising is useful here because, as Pollock defines the term, it draws attention to the transformative work of metrics, analytics, platform features and other devices that re-package social activity through modes of datafication and analysis. The Save the Palace group mobilised in a particular way so as to channel their activities, make them visible and archival, to capture the resonant effects of their public protest through a platform that would best make that public visible to itself. The growth of the interest in the Facebook Page feeds back on itself reflexively as more people encounter it and participate. Contrary to critiques of “clicktivism”, these acts combine digital-material events and activities that were to become consequential for the public protest – such as the engagement activities around the November 2015 event described in Figure 2.In addition, presenting the research in court introduced particular hurdles, in finding “the meaningful data” appropriate to the needs of the case, “visualizing social data for social purposes”, and the need to be “evocative as well as accurate” (Donath, 16). The visualisation and presentation of the data needed to afford a valid and meaningful expression of the social significance the Palace. Which layout algorithm to use? What scale do we want to use? Which community detection algorithm and colour scheme for nodes? These choices involve challenges regarding legibility of visualisations of public data (McCosker and Wilken; Kennedy et al.).The transformative actions at play in these tactics of public data analysis can inform other instances of data-driven protest or social participation, but also leave room for misuse. The interests of developers, for example, could equally be served by monitoring protesters’ actions through the same data, or by targeting disagreement or ambiguity in the data. Similarly, moves by Facebook to restrict access to Page data will disproportionately affect those without the means to pay for access. These tactics call for further work in ethical principles of open data, standardisation and data literacies for the courts and those who would benefit from use of their own public data in this way.ConclusionsWe have argued through the case of the Save the Palace protest that in order to make use of public social media data to define a data public, multiple levels of data literacy, access and affordising are required. Rather than assuming that public data simply constitutes a data public, we have emphasised: a) the consequentiality of the movement; b) sufficient connection over time; and c) affective or emotional qualities of connection and interaction with public events. This includes the activities of the core members of the Save the Palace protest group, and the tens of thousands who engaged in some way with the Page. It also involves Facebook’s data affordances as these allow for the extraction of public data, alongside our choices in analysis and visualisation, and the court’s capacity and openness to accept all of this as indicative of the social value (connections, sentiment, attachment) it sought for the case. The Senior Member and Member presiding over the case had little knowledge of Facebook or other social media platforms, did not use them, and hence themselves had limited capacity to recognise the social and cultural nuances of activities that took place through the Facebook Page. This does not exclude the use of the data but made it more difficult to present a picture of the relevance and consequence of the data for understanding the social value evident in the contested building. 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