Zeitschriftenartikel zum Thema „Trefoil formation“

The limited use of birdsfoot trefoil (Lotus corniculatus L.) in feed production is determined by a large shortage of its seeds. Currently, the supply of certified seeds on the commercial market is about ten times less than the scientifically based volume of the need for the seed material of this crop. The biological feature of the birdsfoot trefoil is the continuous shoot formation and growth during the entire growing season, the extended flowering period. This makes it difficult to determine the optimal timing and methods of harvesting seed stands. Harvesting is associated with large crop losses due to uneven ripening of beans and their strong cracking in hot weather with low relative humidity. The abundance of a large number of green shoots increases the humidity of the grass stand. This is very difficult to thresh and require desiccation. The choice of optimal time and method of threshing is the most difficult stage technology seed production of birdsfoot trefoil. Studies have shown that when 69–93% of the beans were browned, the biological yield reached its maximum values of 308–320 kg/ha. This period in typical weather conditions occurred 35–50 days after the beginning of flowering. During this period, the filling of seeds stopped. Crop performance has reached the standard values. The optimal period of direct threshing with pre-desiccation of the herbage, which provided the highest actual seed collection of 191–206 kg/ha, is the period of browning about 70% of the beans.

In pot trial the biochemical composition and phosphorus use efficiency of birdsfoot trefoil, sainfoin and subterranean clover grown pure and in mixtures with perennial ryegrass in the next ratios were studied in the Institute of Forage Crops, Pleven, Bulgaria: birdsfoot trefoil + perennial ryegrass (50:50%); sainfoin + perennial ryegrass (50:50%); subterranean clover + perennial ryegrass (50:50%); birdsfoot trefoil + subterranean clover + perennial ryegrass (33:33:33%); sainfoin + subterranean clover + perennial ryegrass (33:33:33%). The highest crude protein content was found in the aboveground mass of birdsfoot trefoil (19.17%) and sainfoin (19.30%). The water soluble sugars contents in mixtures was found higher compared to the pure grown legumes. Birdsfoot trefoil showed the highest phosphorus use efficiency for plant biomass accumulation and nodules formation. In mixtures the phosphorus use efficiency was found be higher as compared to the same in pure grown legumes.

In order to optimized the power cable layout design in tunnel, and improved the operation ability of the cable line, this paper analyzed the cable rating and the short-circuit force under different arrangements. 220kV cable circuit was simulated in the tunnel with conditions of the flat and the trefoil arrangements. The temperature and the rating of the 220kV cable under the condition of the two arrangements were calculated in this paper. The short circuit force peak value and its direction of the cable line under short circuit current of 50kA under the condition of the two arrangements were simulated in this paper. The results show that, the cable rating in flat arrangement is better than in trefoil formation. The value and the direction of the short circuit force in tunnel are better in flat arrangement than in trefoil formation. So it is suggested that the flat arrangement is preferred when conditions permit.

RUNX2, a master osteogenic transcript ion factor, is overexpressed in several cancer cells; in melanoma it promotes cells migration and invasion as well as neoangiogenesis. The annual mortality rates related to metastatic melanoma are high and novel agents are needed to improve melanoma patients’ survival. It has been shown that lectins specifically target malignant cells since they present the Thomsen–Friedenreich antigen. This disaccharide is hidden in normal cells, while it allows selective lectins binding in transformed cells. Recently, an edible lectin named BEL β-trefoil has been obtained from the wild mushroom Boletus edulis. Our previous study showed BEL β-trefoil effects on transcription factor RUNX2 downregulation as well as on the migration ability in melanoma cells treated in vitro. Therefore, to better understand the role of this lectin, we investigated the BEL β-trefoil effects in a zebrafish in vivo model, transplanted with human melanoma cells expressing RUNX2. Our data showed that BEL β-trefoil is able to spread in the tissues and to reduce the formation of metastases in melanoma xenotransplanted zebrafish. In conclusion, BEL β-trefoil can be considered an effective biomolecule to counteract melanoma disease.

ABSTRACT The trefoil peptide intestinal trefoil factor (ITF) plays a critical role in the protection of colonic mucosa and is essential to restitution after epithelial damage. These functional properties are accomplished through coordinated promotion of cell migration and inhibition of apoptosis. ITF contains a unique three-looped trefoil motif formed by intrachain disulfide bonds among six conserved cysteine residues, which is thought to contribute to its marked protease resistance. ITF also has a seventh cysteine residue, which permits homodimer formation. A series of cysteine-to-serine substitutions and a C-terminally truncated ITF were made by PCR site-directed mutagenesis. Any alteration of the trefoil motif or truncation resulted in loss of protease resistance. However, neither an intact trefoil domain nor dimerization was required to promote cell migration. This pro-restitution activity correlated with the ability of the ITF mutants to activate mitogen-activated protein (MAP) kinase independent of phosphorylation of the epidermal growth factor (EGF) receptor. In contrast, only intact ITF retained both phosphatidylinositol 3-kinase and the EGF receptor-dependent antiapoptotic effect in HCT116 and IEC-6 cells. The inability to block apoptosis correlated with a loss of trefoil peptide-induced transactivation of the EGF receptor or Akt kinase in HT-29 cells. In addition to defining structural requirements for the functional properties of ITF, these findings demonstrate that distinct intracellular signaling pathways mediate the effects of ITF on cell migration and apoptosis.

The single-domain human trefoil proteins [pNR-2/pS2 and human intestinal trefoil factor (hITF)] have seven cysteine residues, of which six are involved in maintaining the structure of the trefoil domain. The seventh does not form part of the trefoil domain and is located three residues from the C-terminus. The ability of the pNR-2/pS2 single trefoil domain protein to dimerize was examined by using recombinant protein with either a cysteine or a serine residue at this position by equilibrium ultracentrifugation, laser-assisted desorption MS, gel filtration and PAGE. pNR-2/pS2 Cys58 formed dimers, whereas pNR-2/pS2 Ser58 did not. Experiments in which the dimer was treated with thiol agents demonstrated that the dimer was linked via a disulphide bond and that the intermolecular disulphide bond was more susceptible to reduction than the intramolecular disulphide bonds. To examine whether dimeric pNR-2/pS2 was secreted by oestrogen-responsive breast cancer cells, which are known to express pNR-2/pS2 mRNA, conditioned medium was separated on non-denaturing polyacrylamide gels, transferred to PVDF membrane and reacted with antiserum against pNR-2/pS2. Monomeric and dimeric pNR-2/pS2 were detected but the majority of the protein reactivity was associated with a larger protein. Treatment of this protein with thiol agents suggested that it is an oligomer containing pNR-2/pS2 linked to another protein by a disulphide bond. These studies suggest that the biological action of pNR-2/pS2 single-domain trefoil protein might involve the formation of homodimers or oligomers with other proteins.

Background/Aims: Tff3 protein plays a well recognized role in the protection of gastrointestinal mucosa. The role of Tff3 in the metabolism is a new aspect of its function. Tff3 is one of the most affected liver genes in early diabetes and fatty liver rodent models. The aim of this study was to investigate the effect of Tff3 deficiency on lipid and carbohydrate metabolism and on markers of oxidative stress that accompanies metabolic deregulation. Methods: Specific markers of health status were determined in sera of Tff3 deficient mice, including glucose level, functional glucose and insulin tolerance. Composition of fatty acids (FAs) was determined in liver and blood serum by using gas chromatography. Oxidative stress parameters were determined: lipid peroxidation level via determination of lipid hydroperoxide and thiobarbituric acid reactive substances (TBARS), antioxidative capacity (FRAP) and specific antioxidative enzyme activity. The expression of several genes and proteins related to the metabolism of lipids, carbohydrates and oxidative stress (CAT, GPx1, SOD2, PPARα, PPARγ, PPARδ, HNF4α and SIRT1) was determined. Results: Tff3 deficient mice showed better glucose utilization in the glucose and insulin test. Liver lipid metabolism is affected and increased formation of small lipid vesicles is noticed. Formation of lipid droplets is not accompanied by increased liver oxidative stress, although expression/activity of monitored enzymes is deregulated when compared with wild type mice. Tff3 deficient mice exhibit reduced expression of metabolism relevant SIRT1 and PPARγ genes. Conclusion: Tff3 deficiency affects the profile and accumulation of FAs in the liver, with no obvious oxidative stress increase, although expression/activity of monitored enzymes is changed as well as the level of SIRT1 and PPARγ protein. Considering the strong downregulation of liver Tff3 in diabetic/obese mice, presence in circulation and regulation by food/insulin, Tff3 is an interesting novel candidate in metabolism relevant conditions.

The experiment was conducted in the period of 2013-2015 in the experimental field of RIMSA-Troyan. The influence of bio-fertilizers, such as boron humate and molybdenum humate at doses of 1600 ml/ha, was studied. They were applied in the beginning of bud-formation period on legumes, such as bird's-foot-trefoil, sainfoin, and red clover. The highest positive effect of studied bio-fertilizers over productivity was found in the treatment of grassland of sainfoin. Its yield increased, after the application of boron humate, in comparison with the control by 8.3%, and molybdenum humate increased the productivity of red clover by 4.0%. Both bio-fertilizers showed a negative effect in relation to dry matter yield of bird's-foot trefoil. Foliar treatment with boron and molybdenum humate of bird's-foot-trefoil, red clover and sainfoin decreased the level of weed infestation of crops and had an influence over the biological characteristics of grass species and meteorological conditions. As a result of the influence of bio-fertilizers, the stem height of sainfoin and red clover increased, which had a favorable influence over productivity of their grasslands.

The article presents the results of eight-year (2012-2019) field research on the formation of the production process of multicomponent legume and cereal grass stands while simulating pasture use. The comparison study was carried out with the use of basic grass mixtures: meadow clover (Trifolium pratense) VIK 7 + creeping clover (Trifolium repens) VIK 70 + timothy grass (Phleum pretense L.) VIK 9 + meadow fescue (Festuca pratensis) Sakharovskaya. Studied were agrophytocenoses with the use of pasture ryegrass (Lolium perenne) VIK 66 and festulolium VIK 90. As legume components, alfalfa changeable (Medicago varia) Nakhodka, birds-foot trefoil (Lotus corniculatus) Solnyshko and creeping clover VIK 70 were used. To increase the period of use, red fescue (Festuca rubra) Sigma, having the ability to regenerate itself was included into the composition of some mixtures. It has been established that alfalfa changeable, birds-foot trefoil and red fescue are the most adaptive to the conditions of drained lands. The analysis of the results of the study of the botanical composition of grass stands showed that over 8 years of growth, cereal grasses decreased their participation in the composition of herbage from 32.0-47.8 % to 1.8-22.3 % (excluding grass stands with red fescue). Four-component grass stands based on pasture ryegrass and festulolium with the use of timothy grass proved to be the most resistant to preserving sown grass species (46.3-63.7 %) and to weed infestation. Introducing additional legume species (alfalfa changeable and birds-foot trefoil) into the herbal mixture increased the period of use and significantly raised the yield of green mass of grass stands. The highest yield of green mass was obtained by herbage with alfalfa changeable and birds-foot trefoil - 23.7-26.4 t / ha, which in the seventh year of use exceeded the base grass mixture by 3.2-6.5 t / ha or 13.5-24.6 %. The results of fodder evaluation of legume-cereal ryegrass and festulolium grass stands, which denote the high-quality green fodder in conditions of drained soils of the Upper Volga Region, are presented.

The article presents the results of three-year (2017-2019) field studies on changes in agrochemical characteristics of soil and formation of productivity of birdsfoot trefoil and timothy grass mixture in dependence of the liming (hydrolytic acidity 0.25) and application of mineral fertilizers, phosphorite meal in particular. The soil is sod-podzolic, medium loamy, strongly acidic, with intermediate concentration of mobile phosphorous and metabolizable potassium, humus content is 1.9 %, and mobile aluminum – from 4.27 to 5.24 mg/kg of soil. It has been established that application of finely ground lime at a dose of 3 tons per hectare prior to sowing of birdsfoot trefoil and timothy grass mixture decreased soil acidity in the layer of 0-10 cm from 4.2 to 5.6 pH units by next autumn. Application of phosphorite meal (1t/ha) led to an increase in the concentration of mobile phosphorous in the layer of 0-10 cm by 135-163 %. The highest productivity of birdsfoot trefoil and timothy mixture during 3 years was obtained by means of combined application of lime and mineral fertilizers, and for 3 years on the average the yield of metabolic energy exceeded 50 GJ/ha, which is over 35 % higher than in the control variant without fertilizers. The use of mineral fertilizers increased grass mixture productivity only during the first two years. The applied dosage of phosphorite meal (1t/ha) promoted the increase in grass productivity on strongly acidic soil only in the year of sowing. Additional treatment with ammonium nitrate (N30) together with phosphorous and potassium fertilizers (Р60К60), as well as potassium chloride (К60) with phosphorite meal did not result in significant rise in metabolizable energy yield.

TMEM190, a small transmembrane protein containing the trefoil domain, was previously identified by our proteomic analysis of mouse sperm. Two structural features of TMEM190, ‘trefoil domain’ and ‘small transmembrane protein’, led us to hypothesize that this protein forms a protein–protein complex required during fertilization, and we characterized TMEM190 by biochemical, cytological, and genetic approaches. We showed in this study that the mouse Tmem190 gene exhibits testis-specific mRNA expression and that the encoded RNA is translated into a 19-kDa protein found in both testicular germ cells and cauda epididymal sperm. Treatment of the cell surface with proteinase K, subcellular fractionation, and immunofluorescence assay all revealed that mouse TMEM190 is an inner-acrosomal membrane protein of cauda epididymal sperm. During the acrosome reaction, TMEM190 partly relocated onto the surface of the equatorial segment, on which sperm–oocyte fusion occurs. Moreover, TMEM190 and IZUMO1, which is an immunoglobulin-like protein required for gamete fusion, co-localized in mouse sperm both before and after the acrosome reaction. However, immunoprecipitates of TMEM190 contained several sperm proteins, but did not include IZUMO1. These findings suggest that a mouse sperm protein complex(es) including TMEM190 plays an indirect role(s) in sperm–oocyte fusion. The role(s), if any, is probably dispensable since Tmem190-null male mice were normally fertile.

Three-phase underground power cables can induce voltages and currents in their recover sheaths. The induced voltages and currents in sheath are undesirable. They generate heat losses and reduce the cable ampacity. Moreover, the induced voltages can generate electric shock to the workers, who maintain the power cables. It is very important to predict the sheath voltages and currents, which depend on different parameters, such as the sheath grounding system, the geometry of the cables, the gap between them, etc. In this paper, the voltages and currents induced in sheath for different installation of underground power cables (trefoil and flat formation) are studied and presented. For each case study of installation, the results of sheath voltages and currents have been computed and compared. Finally, the case of cross bonding with increasing of cable spans can reduce the sheath voltage significantly.

We investigate aspects of topology in protein folding. For this we numerically simulate the temperature driven folding and unfolding of the slipknotted archaeal virus protein AFV3-109. Due to knottiness the (un)folding is a topological process, it engages the entire backbone in a collective fashion. Accordingly we introduce a topological approach to model the process. Our simulations reveal that the (un)folding of AFV3-109 slipknot proceeds through a folding intermediate that has the topology of a trefoil knot. We observe that the final slipknot causes a slight swelling of the folded AFV3-109 structure. We disclose the relative stability of the strands and helices during both the folding and unfolding processes. We confirm results from previous studies that pointed out that it can be very demanding to simulate the formation of knotty self-entanglement, and we explain how the problems are circumvented: The slipknotted AFV3-109 protein is a very slow folder with a topologically demanding pathway, which needs to be properly accounted for in a simulation description. When we either increase the relative stiffness of bending, or when we decrease the speed of ambient cooling, the rate of slipknot formation rapidly increases.

Trefoil factor family peptides (TFF1, TFF2, TFF3) are typically co-secreted together with mucins. Tff1 represents a gastric tumor suppressor gene in mice. TFFs are also synthesized in minute amounts in the immune and central nervous systems. In mucous epithelia, they support rapid repair by enhancing cell migration (“restitution”) via their weak chemotactic and anti-apoptotic effects. For a long time, as a paradigm, this was considered as their major biological function. Within recent years, the formation of disulfide-linked heterodimers was documented for TFF1 and TFF3, e.g., with gastrokine-2 and IgG Fc binding protein (FCGBP). Furthermore, lectin activities were recognized as enabling binding to a lipopolysaccharide of Helicobacter pylori (TFF1, TFF3) or to a carbohydrate moiety of the mucin MUC6 (TFF2). Only recently, gastric TFF1 was demonstrated to occur predominantly in monomeric forms with an unusual free thiol group. Thus, a new picture emerged, pointing to diverse molecular functions for TFFs. Monomeric TFF1 might protect the gastric mucosa as a scavenger for extracellular reactive oxygen/nitrogen species. Whereas, the TFF2/MUC6 complex stabilizes the inner layer of the gastric mucus. In contrast, the TFF3–FCGBP heterodimer (and also TFF1–FCGBP) are likely part of the innate immune defense of mucous epithelia, preventing the infiltration of microorganisms.

The ruminal fungi Neocallimastix frontalis RE1, Neocallimastix patriciarum 27, Piromyces communis 22, and Orpinomyces joyonii 19-2 were examined for their ability to digest filter paper in the presence of condensed tannins from birdsfoot trefoil (Lotus corniculatus L.). For all four fungi, inhibition of endoglucanases was evident at 100 μg condensed tannins∙mL−1 with nearly complete inhibition at 300 μg condensed tannins∙mL−1. At 100 and 200 μg condensed tannins∙mL−1, the endoglucanase activity of N. frontalis RE1 was greater (P < 0.01) than that of the other three fungal species. Exposure to 100 μg condensed tannins∙mL−1 did not affect the ability of N. frontalis RE1 or N. patriciarum 27 to digest filter paper, and although digestion was reduced, N. frontalis RE1 and P. communis 22 solubilized more than 20% of the filter paper at 500 μg condensed tannins∙mL−1. In contrast, O. joyonii 19-2 was virtually unable to digest filter paper at 300 μg condensed tannins∙mL−1. Mycelia of fungi grown with condensed tannins were covered by filamentous material, which may have arisen from the formation of condensed tannin–protein complexes. Less than 86% of the condensed tannins (as measured by the H2SO4 method) were recovered after 120 h of incubation with N. frontalis RE1, P. communis 22, and N. patriciarum 27. The need for detailed studies to examine the ability of ruminal fungi to metabolize condensed tannins is evident.Key words: ruminal fungi, condensed tannins, cellulose digestion, ruminant.

In this paper, a Frequency-Dependent Pi Model (FDPi) of a three-core submarine cable is presented. The model is intended to be used for the representation of submarine cables in an Offshore Wind Power Plant (OWPP) scenario for both time and frequency domain analysis. The frequency-dependent variation of each conductive layer is modeled by a Foster equivalent network whose parameters are tuned by means of Vector Fitting (VF) algorithm. The complete formulation for the parameterization of the model is presented in detail, which allows an easy reproduction of the presented model. The validation of the model is performed via a comparison with a well-established reference model, the Universal Line Model (ULM) from PSCAD/EMTDC software. Two cable system case studies are presented. The first case study shows the response of the FDPi Model for a three-core submarine cable. On the other hand, the second case study depicts the response of three single-core underground cables laying in trefoil formation. This last case shows the applicability of the FDPi Model to other types of cable systems and indirectly validates the response of the aforementioned model with experimental results. Additionally, potential applications of the FDPi model are presented.

<p>The cofactor of BRCA1 (COBRA1), which also refers to negative elongation factor polypeptide B (NELF-B), is a negative elongation factor (NELF) subunit that has been implicated in the development and progression of several cancers. While reduced COBRA1 expression has been associated with metastatic breast cancer, COBRA1 negatively regulates the activator protein-1 (AP-1) complex, leading to the down-regulation of trefoil factor-1 (TFF1) expression in gastric cancer cell lines. The involvement of COBRA1 in hepatocellular carcinoma (HCC) tumor formation and progression is not known. Here, we investigated the expression of COBRA1, the AP-1 complex, and TFF1 in several HCC cell lines; ranging from low- to high-grade HCC cell lines generated from patients with different stages of the disease. Our results showed that the COBRA1 protein was highly expressed in the low-grade HCC cell line, while significantly down-regulated in high-grade HCC cell lines. TFF1, previously regarded as a COBRA1 target gene, was only expressed in the low-grade HCC cell line and the control cells. Our results suggest that COBRA1 may play a role in HCC pathogenesis and progression. The TFF1 mRNA expression profile was uncorrelated to that of the AP-1 complex subunit proteins, which suggests the involvement of other regulatory pathways in TFF1 expression; however, this requires further study.</p>

Trefoil factor family peptide 3 (TFF3) is supposed to have tumor suppressive functions in retinoblastoma (RB), but the functional pathway is not completely understood. In the study presented, we investigated the downstream pathway of TFF3 signaling in Y79 RB cells. Results from pG13-luciferase reporter assays and western blot analyses indicate induced p53 activity with an upregulation of miR-34a after TFF3 overexpression. Expression levels of the predicted miR-34a target epithelial membrane protein 1 (EMP1) are reduced after TFF3 overexpression. As revealed by WST-1 assay, BrdU, and DAPI cell counts viability and proliferation of Y79 cells significantly decrease following EMP1 knockdown, while apoptosis levels significantly increase. Opposite effects on Y79 cells’ growth could be shown after EMP1 overexpression. Caspase assays showed that EMP1 induced apoptosis after overexpression is at least partially caspase-3/7 dependent. Colony formation and soft agarose assays, testing for anchorage independent growth, revealed that EMP1 overexpressing Y79 cells have a significantly higher ability to form colonies. In in ovo chicken chorioallantoic membrane (CAM) assays inoculated EMP1 overexpressing Y79 cells form significantly larger CAM tumors. Moreover, miR-34a overexpression increases sensitivity of Y79 cells towards RB chemotherapeutics, however, without involvement of EMP1. In summary, the TFF3 signaling pathway in Y79 RB cells involves the activation of p53 with downstream induction of miR-34a and subsequent inhibition of EMP1.

ABSTRACTMucin glycoproteins are secreted in large amounts by the intestinal epithelium and constitute an efficient component of innate immune defenses to promote homeostasis and protect against enteric pathogens. In this study, our objective was to investigate how the bacterial enteropathogenShigella flexneri, which causes bacillary dysentery, copes with the mucin defense barrier. We report that uponin vitroinfection of mucin-producing polarized human intestinal epithelial cells, virulentS. flexnerimanipulates the secretion of gel-forming mucins. This phenomenon, which is triggered only by virulent strains, results in accumulation of mucins at the cell apical surface, leading to the appearance of a gel-like structure that favors access of bacteria to the cell surface and the subsequent invasion process. We identify MUC5AC, a gel-forming mucin, as a component of this structure. Formation of this gel does not depend on modifications of electrolyte concentrations, induction of trefoil factor expression, endoplasmic reticulum stress, or response to unfolded proteins. In addition, transcriptional and biochemical analyses of infected cells reveal modulations of mucin gene expression and modifications of mucin glycosylation patterns, both of which are induced by virulent bacteria in a type III secretion system-dependent manner. Thus,S. flexnerihas developed a dedicated strategy to alter the mucus barrier by targeting key elements of the gel-forming capacity of mucins: gene transcription, protein glycosylation, and secretion.

In Australia in 2011 the Federal government introduced a voluntary offset scheme called the Carbon Farming Initiative, which allows farmers to receive carbon credits when they reduce or sequester greenhouse gas emissions. Various mitigation options have since been explored for their potential to reduce on-farm greenhouse gas emissions. Among these is the use of alternative pastures that lower methane (CH4) production of grazing animals such as Lotus corniculatus, a legume that contains condensed tannins that inhibit the formation of CH4 in the rumen. Lotus has other benefits for sheep production such as increased wool growth, liveweight gain and fecundity. This study modelled the potential emission, production and economic outcomes for wool and lamb enterprises that incorporate lotus in their pastures, evaluating the impact of existing farm productivity, lotus intake and carbon price. Depending on the amount of lotus consumed and the CH4 reduction rate, CH4 emissions fell by 0.02–0.38 t carbon dioxide equivalents (CO2e)/ha and 0.05–0.48 t CO2e/ha on wool and prime lamb enterprises, respectively. At a price of $6/t CO2e potential offset income attributable to use of lotus across all enterprises was $0.12–2.91/ha. Increases in income from increased productivity were 15–30 times greater than from potential carbon offset income. Income was driven by the amount of lotus dry matter intake and the subsequent production benefits. Over a 10-year period prime lamb enterprises generated up to $50/ha in profit by using lotus, due to increased liveweight gain and fecundity. In most modelled scenarios wool enterprises would not cover the cost of lotus pasture establishment. This research demonstrated that 18–23% and 37–46% of lotus intake within the diet was required to generate production enough to cover pasture establishment costs on prime lamb and wool enterprises, respectively. Potential carbon offset income would not be sufficient for farmers to establish lotus without the productivity benefits. While extra profit may be gained on prime lamb enterprises through the use of lotus, problems with persistence must first be overcome for lotus to be adopted on farms.

Endolysosomes are key players in cell physiology, including molecular exchange, immunity, and environmental adaptation. They are the molecular targets of some pore-forming aerolysin-like proteins (ALPs) that are widely distributed in animals and plants and are functionally related to bacterial toxin aerolysins. βγ-CAT is a complex of an ALP (BmALP1) and a trefoil factor (BmTFF3) in the firebelly toad (Bombina maxima). It is the first example of a secreted endogenous pore-forming protein that modulates the biochemical properties of endolysosomes by inducing pore formation in these intracellular vesicles. Here, using a large array of biochemical and cell biology methods, we report the identification of BmALP3, a paralog of BmALP1 that lacks membrane pore-forming capacity. We noted that both BmALP3 and BmALP1 contain a conserved cysteine in their C-terminal regions. BmALP3 was readily oxidized to a disulfide bond-linked homodimer, and this homodimer then oxidized BmALP1 via disulfide bond exchange, resulting in the dissociation of βγ-CAT subunits and the elimination of biological activity. Consistent with its behavior in vitro, BmALP3 sensed environmental oxygen tension in vivo, leading to modulation of βγ-CAT activity. Interestingly, we found that this C-terminal cysteine site is well conserved in numerous vertebrate ALPs. These findings uncover the existence of a regulatory ALP (BmALP3) that modulates the activity of an active ALP (BmALP1) in a redox-dependent manner, a property that differs from those of bacterial toxin aerolysins.

Increased expression of trefoil factor 3 (TFF3) has been reported in colorectal carcinoma (CRC), being correlated with distant metastasis and poor clinical outcomes. Amongst the CRC subtypes, mesenchymal (CMS4) CRC is associated with the worst survival outcome. Herein, the functional roles of TFF3 and the pharmacological inhibition of TFF3 by a novel specific small molecule TFF3 inhibitor—2-amino-4-(4-(6-fluoro-5-methylpyridin-3-yl)phenyl)-5-oxo-4H,5H-pyrano[3,2-c]chromene-3-carbonitrile (AMPC) in CMS4 CRC was explored. Forced expression of TFF3 in CMS4 CRC cells promoted cell proliferation, cell survival, foci formation, invasion, migration, cancer stem cell like behaviour and growth in 3D Matrigel. In contrast, siRNA-mediated depletion of TFF3 or AMPC inhibition of TFF3 in CMS4 CRC cells decreased oncogenic behaviour as indicated by the above cell function assays. AMPC also inhibited tumour growth in vivo. The TFF3-stimulated oncogenic behaviour of CMS4 CRC cells was dependent on TFF3 activation of the p44/42 MAPK (ERK1/2) pathway. Furthermore, the forced expression of TFF3 decreased the sensitivity of CMS4 CRC cells to 5-fluorouracil (5-FU); while depleted TFF3 expression enhanced 5-FU sensitivity in CMS4 CRC cells. 5-FU treatment induced TFF3 expression in CMS4 CRC cells. AMPC, when used in combination with 5-FU in CMS4 CRC cells exhibited a synergistic inhibitory effect. In summary, this study provides functional evidence for TFF3 as a therapeutic target in CMS4 CRC.

Arginine residues in barley alpha-amylase/subtilisin inhibitor (BASI) involved in binding to barely alpha-amylase 2 (AMY2) were differentially labelled using AMY2 as protectant and phenylglyoxal (PGO) and [14C]PGO as modifying agents. Chymotryptic fragments of labelled BASI were purified by reverse-phase HPLC, and we concluded that the radiolabelled Arg-27, Arg-155 and most likely Arg-127, identified by amino acid, sequence and 14C analyses, are protected by AMY2. While Arg-106 and Arg-107 showed intermediate reactivity and apparently were only partly accessible, Arg-15, Arg-41 and Arg-61 reacted with PGO and were thus exposed in the BASI-AMY2 complex. Patterns of arginine modification by [14C]PGO in free or in AMY2-complexed BASI were consistent with the results of differential labelling. The AMY2-protected arginines in BASI are at a distance from each other, as deduced from crystal structures of different beta-trefoil proteins (Erythrina caffra and soybean trypsin inhibitors, interleukin-1 alpha and -1 beta and WASI, the wheat homologue), suggesting that the BASI-AMY2 complex has multiple contacts at a larger interface. Accordingly, 11-16-residue-long BASI oligopeptides synthesized to include Arg-27, Arg-106/Arg-107 or Arg-127 were unable to suppress the formation of BASI-AMY2 or the effect of an inhibitory monoclonal antibody to BASI. Since Arg-27 is not conserved in rice and wheat ASIs, we further propose that Arg-155 in BASI is the kinetically identified PGO-sensitive group that is essential for inhibition [Abe, Sidenius and Svensson (1993) Biochem. J. 293, 151-155].

1. The efficient repair of gastrointestinal mucosal injuries is essential in the preservation of the epithelial barrier to luminal antigens. Accumulated evidence suggests that epithelial migration plays a major part in this repair by rapidly resealing defects induced by both physiological and pathological insults, a process termed restitution. 2. This migration has been modelled in various ways, most commonly in mechanically wounded monolayers of cell lines or cells in primary culture, and in wounded human or animal tissue. Evidence from these models indicates that migration is a highly complex process, which is likely to involve the tightly controlled spatial and temporal interaction of multiple factors: (i) extracellular molecules such as soluble factors (e.g. growth factors, trefoil peptides, cytokines) and matrix components (e.g. collagen, laminin, fibronectin); (ii) signalling molecules activated by the interaction of these factors with cell surface receptors (e.g. protein kinases, phospholipases, low-molecular-weight GTPases); (iii) factors which regulate adhesion to other cells (e.g. E-cadherin) and to matrix components (e.g. integrins, hyaluronic acid receptors); (iv) factors which regulate detachment from the extracellular matrix (e.g. urokinase-type plasminogen activator, matrix metalloproteinases); and (iv) molecules which regulate cytoskeletal function (e.g. Rac), which allows the formation of specialized cellular processes termed lamellipodia. 3. The identification of physiologically relevant factors that stimulate epithelial cell migration, and a better understanding of their mechanism of action, may be beneficial in the development of novel therapeutic approaches in diseases such as inflammatory bowel disease through the pharmacological or dietary enhancement of this migration.

AbstractTrefoil factor family 3 (Tff3) peptide is present during intrauterine endochondral ossification in mice, and its deficiency affects cancellous bone quality in secondary ossification centers of mouse tibiae. The aim of this study was to quantitatively analyze parameters describing the growth plate and primary ossification centers in tibiae of 1-month-old wild-type and Tff3 knock-out mice (n=5 per genotype) by using free and open-source software. Digital photographs of the growth plates and trabecular bone were processed by open-source computer programs GIMP and FIJI. Histomorphometric parameters were calculated using measurements made with FIJI. Tff3 knock-out mice had significantly smaller trabecular number and significantly larger trabecular separation. Trabecular bone volume, trabecular bone surface, and trabecular thickness showed no significant difference between the two groups. Although such histomorphological differences were found in the cancellous bone structure, no significant differences were found in the epiphyseal plate histomorphology. Tff3 peptide probably has an effect on the formation and quality of the cancellous bone in the primary ossification centers, but not through disrupting the epiphyseal plate morphology. This work emphasizes the benefits of using free and open-source programs for morphological studies in life sciences.

The number of people with inflammatory bowel disease (IBD) is constantly increasing worldwide. The main factors that have effects on the etiology of the disease are genetic, environmental and immunological. However, the mechanism of disease development and effective treatment of IBD have not yet been found. Animal models help address these problems. The most popular model is considered to be transgenic models in which individual genes are knocked out. One of such models for the study of IBD are mice with a null mutation of theMuc2gene encoding the Mucin-2 protein, which is involved in the formation of a protective mucin layer in the small and large intestine. Some of transcription factors that change the expression of intestinal genes are involved in the development of IBD and colorectal cancer. One of such transcription factors is “zinc fnger” domain-containing protein Kaiso which is able to bind to methylated DNA. In this study, we assessed the role of Kaiso in the development of intestinal inflammation using the experimental model of C57BL/6Muc2-/-Kaiso-/-. We have shown that mice with impaired intestinal barrier function that develop processes similar to human IBD also develop inflammatory responses, such as increased expression ofIl1,TnfandIl17agenes. The defciency of the Kaiso transcription factor in Mucin-2 knockout mice causes a decrease in the expression level of only theCox2andTff3genes. Perhaps a decline in the expression of the gene encoding cyclooxygenase-2 can lead to a decrease in the expression of the antibacterial factor Trefoil factor 3. However, in the experimental model of IBD, Kaiso protein did not play a signifcant role in the regulation of pro-inflammatory cytokines of tumor necrosis factor and interleukins 1 and 17.

Abstract Background The high rate of cell turnover in the intestinal epithelium is supported by the LGR5+ crypt base columnar (CBC) stem cells, which are located at the lower part of the gland. Among of the various factors and signals like Wnt and Notch, YAP1 (yes associated protein) also plays an important role in stemness of CBC stem cells. YAP1 is the effector of the Hippo pathway. Hippo Pathway restricts the cells proliferation, tissues overgrowth and cancer formation through the phosphorylation and inactivation of the YAP1 protein. When active, YAP1 transfers into nucleus, forms the complex with TEADs transcription factors and promotes the transcription of genes involved in cell growth and proliferation. Aims In the present study, we investigated the role of the YAP1 in the colorectal cancer multipotent HT29 cell line, which contain cancer stem cells (CSC). Methods For approaching to this goal, YAP1 expression was knocked down using shRNAs in HT29 cells. Then stem cells and intestinal cell lineages (secretory goblet, Paneth and enteroendocrine and absorptive) markers expression was analyzed using qPCR and Western blot. Results The results showed the reduction of the expression of stem cells markers including LGR5 in YAP1 knockdown HT29 cells compare with control. Expression of the goblet cells markers (MUC2 and trefoil factor 3) and absorptive cells markers (sucrase-isomaltase and dipeptidylpeptidase IV) were significantly increased in YAP1 knockdown cells but Paneth (DEFA5 and lysozyme) and enteroendocrine (CHGA) were not detected. Finally, examination of the main transcription factors for intestinal differentiation revealed an increase in CDX2 expression. Conclusions These results suggest that YAP1 is involved in the maintenance of colorectal cancer stem cells while preventing intestinal differentiation in both secretory and absorptive lineages through the repression of CDX2. Funding Agencies CIHR

The IP3R [IP3 (inositol 1,4,5-trisphosphate) receptor] is responsible for Ca2+ release from the ER (endoplasmic reticulum). We have been working extensively on the P400 protein, which is deficient in Purkinje-neuron-degenerating mutant mice. We have discovered that P400 is an IP3R and we have determined the primary sequence. Purified IP3R, when incorporated into a lipid bilayer, works as a Ca2+ release channel and overexpression of IP3R shows enhanced IP3 binding and channel activity. Addition of an antibody blocks Ca2+ oscillations indicating that IP3R1 works as a Ca2+ oscillator. Studies on the role of IP3R during development show that IP3R is involved in fertilization and is essential for determination of dorso-ventral axis formation. We found that IP3R is involved in neuronal plasticity. A double homozygous mutant of IP3R2 (IP3R type 2) and IP3R3 (IP3R type 3) shows a deficit of saliva secretion and gastric juice secretion suggesting that IP3Rs are essential for exocrine secretion. IP3R has various unique properties: cryo-EM (electron microscopy) studies show that IP3R contains multiple cavities; IP3R allosterically and dynamically changes its form reversibly (square form–windmill form); IP3R is functional even though it is fragmented by proteases into several pieces; the ER forms a meshwork but also forms vesicular ER and moves along microtubules using a kinesin motor; X ray analysis of the crystal structure of the IP3 binding core consists of an N-terminal β-trefoil domain and a C-terminal α-helical domain. We have discovered ERp44 as a redox sensor in the ER which binds to the luminal part of IP3R1 and regulates its activity. We have also found the role of IP3 is not only to release Ca2+ but also to release IRBIT which binds to the IP3 binding core of IP3R.

The phenomenon of early Christian triconchal churches on the Adriatic has already been noted in the scholarly literature. A separate study ‘Le basiliche cruciformi nell’area adriatica’ was published by S. Piussi in 1978, followed by N. Cambi with the 1984 publication ‘Triconchal churches on the Eastern Adriatic’. However, both scholars include triconchal churches in the typological group of ‘cruciform basilicas’ or treat them together with the churches which have three apses with spaces between them placed along the nave. However, because of their specific morphology consisting of the closely placed conchs and a large number of such examples in the Adriatic area, it seems justified to treat them as a separate typological group. These churches had originally been funerary chapels, but many of them subsequently grew into congregational spaces with complex liturgical functions. In addition, among the triconchal churches it is possible to discuss separately the type of a small triconchal cella without a nave, but sometimes provided with a narthex, as form which is different from similar chapels with a long entrance arm in front of the sanctuary. Based on this difference, it is possible to establish a different terminology which classifies cella trichora as the simple trefoil type, and triconchal churches as the more complex type. The latter is relatively numerous in the territory of late antique Dalmatia. The title of this paper stems from those buildings. However, they originate in cellae trichorae. Thus, in the introductory section I am discussing examples of these cellae in the Adriatic and the connection between their appearance and funerary traditions in the Mediterranean in general. The beginnings of Christian funerary architecture in Dalmatia are found in the grouping of round cellae in the cemeteries of ancient Salona, as known from N. Duval’s works, and in the presence of conchs next to the memorial chapel at Muline which was studied by M. Suić. I deem that the early Christian triconchal churches were created through the crystallisation of the forms present in the groups of funerary cellae in such complexes; cella trichora being the simplest form and triconchal church a more complex one. However, both are generically tied to the Roman tradition in pagan and early Christian funerary architecture. On the other hand, early Christian trefoil structures in the majority of examples stand next to the rustic villa which in itself speaks in favour of a private funerary function. Thus, it is important to assume that cellae trichorae and triconchal churches in the beginning represent early Christian memorial chapels, independent of the subsequent development of the complexes which enveloped them.Thus, the memorial chapel at Muline on the island of Ugljan is part of a larger funerary complex. It is still the most thoroughly researched group of early Christian buildings erected next to a Roman rustic villa in Dalmatia. Apart from a similar example at Brijuni, the Muline complex is crucial for the consideration and interpretation of the origins and development of Christianity in late antique rural areas on the Croatian coast of the Adriatic. It reflects the developed Christianity in the urban setting of Zadar. The owner of the villa was obviously a rich citizen who had a memorial chapel erected on his estate for a deceased person about whom we know nothing. The chapel nave is square. Two deep semicircular apses are found at the back; in the southern one was a sarcophagus. The second sarcophagus was buried under the pavement in the nave. Next to the façade was a protyron, a vestibule with a porch resting on two columns. A courtyard was subsequently added in front of the façade and provided with additional cellae around it. According to Suić’s analysis, it seems that the first layer of the memorial chapel was built in the fourth century. At that time it lacked a crystallized form of somewhat later triconchal churches on the Adriatic. Two original conchs at the back stand slightly apart. The third cella next to the back was subsequently added to the north wall. It has a rectangular ground plan similar to those around the courtyard. All this speaks in favour of a gradual multiplication of cellae around the original memorial, a process similar to that at the cemeteries in Salona. In this paper, I am discussing the phenomenon of early Christian and early medieval triconchal churches on the Adriatic. In doing so, I am only considering those which have three conchs along the sanctuary wall. Based on their form, function and date, I classify them into five groups.The first group one consists of relatively early, small cellae trichorae. They had originally been funerary chapels on private estates. The remains of these memorial chapels have been preserved in various locations along the Adriatic coast: from those at Concordia Sagittaria near Aquileia, Betika near Pula, to those at Gata near Salona and Doljani near Duklja. Older examples have been dated to the late fourth or to the first half of the fifth century, which seems to be the date of the formation of this type of Christian memorial.In the second group are somewhat more complex triconchal churches which, unlike the cellae, have a long nave in front of the sanctuary. They are found in the territory of the Roman Dalmatia and therefore referred to by the author as Dalmatian. Unlike the cellae trichorae, which in their original form do not have a long entrance arm preceding the sanctuary conchs like a nave, triconchal churches are characterised by this very element in the front part of the chapel. In this respect they are spatially more developed than the basic, cella trichora type, and thus probably represent a somewhat later variants of trefoil memorial chapels. It seems that the triconchal churches at Dalmatia were mostly built by the late fifth century or in the early sixth century.The third group consists of those churches from the second group which were transformed from the initial funerary chapels into complex triconchal basilicas. Similar to other types of original memorial chapels which were subsequently transformed into congregational churches in Dalmatia, these too were remodelled in mid-sixth century. Thus, by being enveloped by a ring of subsequently added rooms, some triconchal churches were transformed from the original memorial chapels into public congregational churches furnished with liturgical annexes, among which were baptisteries. Baptisteries in particular witness about the nature of the remodelled triconchal churches and newly created complexes, with a trefoil structure at the core. They indicate an increase in conversion of the population which probably caused the building of such structures. Of course, a similar development was shared by other types of originally private chapels in the time when churches were being built after the model of complex basilicas. However, in Dalmatia, there are no examples of such buildings before the age of Justinian i.e. before the second third of the sixth century. It is likely that the mentioned conversion occurred in this period. With it, many older churches, including triconchal churches, became cores of new complexes. Based on the examples of such a development, it is possible to speak convincingly of pre-Justinianic origins of the initial form of Dalmatian triconchal churches.The fourth group is formed by pre-Romanesque triconchal churches. Their morphology differs from early Christian triconchal churches, and they are represented by two subgroups of interesting early medieval churches in Dalmatia. In the first one are numerous centrally-planned buildings while in the second are two longitudinal structures. Both subgroups are characterised by a sanctuary with three semicircular apses. In the centrally-planned buildings they are placed radially and their axes originate at the centre of the rotunda. Thus, they were not arranged in a cruciform way towards the sanctuary as it had regularly been the case in early Christian cellae trichorae or triconchal churches, where the axes of the lateral apses are perpendicular to the axis of the central apse. However, the three conchs grouped at the sanctuary are a crucial spatial feature in the buildings of the first subgroup so, in principle, they can be referred to as triconchal structures. In this group are the church of Holy Trinity at Zadar and a number of Dalmatian hexaconchal churches, as well as the rotunda at Ošlje. In the second subgroup are the longitudinal churches of Holy Saviour at Vrh Rika near Cetina and the church at Lopuška glavica, both near Knin. These two churches have a long nave in front of the sanctuary, and three conchs along the sanctuary wall, as was the case with early Christian triconchal churches. However, the axes of the lateral conchs are not perpendicular to the axis of the main apse but are placed radially. The nave in the church is significantly wider than the diameter of the main apse. The original layout of the church of St Donatus at Zadar, as a free-standing rotunda, was probably created in the in the eighth century. All other pre-Romanesque triconchal churches in Dalmatia have been convincingly dated to the period between the mid-ninth century to the early decades of the tenth century.Finally, the fifth group consist of the Romanesque trefoil churches. These are small, cruciform cellae which have a short entrance arm at the front and three conchs grouped around the core at the back. The front usually rectangular and the conchs are semicircular. They are vaulted with semi-domed vaults. Above the core is a round drum with a dome. Two of those cellae are almost completely preserved and of particular interest due to the intersecting vault ribs below their domes. Stylistic characteristics of these buildings indicate the early Romanesque architectural features of the twelfth century. All other medieval triconchal churches in this group probably also belong to the wider Romanesque period.Finally, regardless of all similar spatial forms in antique and late antique secular buildings, it should be pointed out that the cellae trichorae and triconchal churches originated as Christian memorial chapels, inspired by the gglomerations of the earliest funerary a chapel installed in early Christian cemeteries. The triconchal shape of these chapels originated in these agglomerations and remained related to the funerary and memorial character. It can be concluded that the triconchal churches in Dalmatia were formed with relation to that character and that they persisted from the early Christian time to the mature middle ages. Perhaps it might be naive and mistaken to interpret the morphology of later buildings as being directly influenced by the earlier. Pre-Romanesque rotundas display a variety of triconchal forms which were not known in early Christian architecture of Dalmatia (except the hexaconchal interior of Zadar Baptistery). Nonetheless, polyconchal spaces of early medieval memorial buildings were furnished with a triconchal sanctuary of the same shape as those in early Christian triconchal buildings, and witness about the funerary function in the pre-Romanesque period. The Romanesque trefoil churches, however, recreated the original type, not as direct replicas of early Christian triconchal forms, but through their function, while their shape grew out of the reformation spirit of the great church reform in the Romanesque period. Thus, Dalmatian triconchal churches illustrate a continuous need for private memorial chapels which does not necessarily have to be triconchal but this particular shape has been discussed here because of its peculiarity. Already in the early Christian period, some trefoil structures outgrew their function of a family chapel to become churches for a larger community. That is why they were accompanied by additional liturgical functions and annexes necessary for monastic or parish churches. By this, they were transformed into complex basilicas with additional spaces while the original triconchal structure, situated at the centre, became the church, quadratum populi, sometimes surrounded by a series of interconnected rooms which served as an ambulatory. This might point to the possibility that in some cases the old funerary function of the original memorial chapel could have continued together with the new liturgical rites in the newly formed complex basilica as a congregational church. These changes did not take place in the medieval memorial structures although some hexaconchal churches and the octaconchal church at Ošlje were provided with new annexes soon after the initial building phase, and that added to the rotunda of St Donatus at Zadar included a gallery.

The paper seeks to outline the overall framework for the reception of St. Clement’s tradition in Slavic literacy in northern, Serb-populated areas; the paper also analyzes major Serbian literary monuments, both Glagolitic and Cyrillic, which may be brought into a close relationship with the literacy tradition of St. Clement. They are presented individually, also taking into account an earlier linguistic background from which they stemmed. These older linguistic traits which are Old Slavonic as well as some later characteristics are generally possible to arrange in an ideal chronological sequence. This makes it possible to suggest a relative chronology of the formation of some Serbian literary monuments. There are also some local linguistic traits and other parameters that allow one to date Serbian literary monuments more precisely and, sometimes, even to delimit their territory of origin. This series begins with the Codex Marianus and continues with Miroslav’s Gospel, the Mihanović Fragment, the Gršković Fragment, Bratko’s Menaion, the Jerusalem Palimpsest, and the Belgrade Prophetologion, ending with the Serbian Prophetologion from St. Petersburg and Kiev. One must keep in mind that the Serbian language, which underlies the spoken background of the Serbian redaction of the Church Slavonic language, was, shortly after its formation (up to the end of the 11th century), still dialectically undiversified (regardless of the potentially heterogeneous situation before the 9th century); thus, based on the current body of knowledge, it is not possible to identify dialectical traits that would provide more specific information about individual writings. However, traces of the general logic of the developmental dynamics of the folk language can be identified in the language of the only 11th-century source presented in this paper: the Codex Marianus. This literary monument is temporally and spatially located in the third quarter of the 11th century and the southeastern boundary of Raška (roughly in Poibarje), near the fortress of Zvečan and the early medieval settlement of Čečan. Miroslav’s Gospel is dated to the period between 1161 and 1170 (ca. 1165) and is linguistically associated with the territory of the Bishopric of Raška because its scribes were the bearers of a dialect typical of this region: the manuscript either originates from Raška or it was written by Rascian scribes in some other area. Based on a rather large number of literary monuments, it is possible to get insight into the third stage in the life of this form of literacy in Polimlje, where the hereditary estates of the Nemanjićs and their relatives were located. From the early Middle Ages this area witnessed lively ecclesiastical activities, though they were based on the Roman Rite. One of the cultural centers must have been located around the trefoil church of St. John at Zaton (9th–11th centuries). In this wider area, a more conservative Serbian literary tradition, which can be traced in the Mihanović Fragment, could have persisted slightly longer. The Mihanović Fragment was the purest representative of the Serbian redaction, without secondary shadings typical of the innovative southern Slavic areas in the 11th century (with the mildest divergence from the vernacular variety when pronouncing the literary language), and it was still based on the linguistic background shaped by St. Clement. The linguistic picture of this literary monument indicates that it could have originated from an area where an ancient linguistic redaction dating back to the early 10th century, or perhaps an even older variety of a literary language from the 9th century (associated with the Roman Rite) combined with a later South Slavic layer of undetermined age (10th–11th centuries), persisted.

Кариес является одной из актуальной проблем современной стоматологии и занимает первое место по частоте и интенсивности среди всех стоматологических заболеваний. Установлено, что поражение зубов кариесом - это заключительный этап взаимодействия многих эндогенных и экзогенных факторов, индивидуальных для каждого пациента. Большинство научно-исследовательских работ по изучению причин возникновения кариеса зубов доказывают, что непосредственным этиологическим фактором является локальное изменение pH под биопленкой, фиксированной на поверхности зуба, вследствие расщепления углеводов микроорганизмами и образования органических кислот. Следует упомянуть, что трилистник Кейза служит графической моделью теории возникновения кариеса в современной стоматологии, в которой равноценны три состояния, а именно кариесогенная флора, легкоферментируемые углеводы, низкая кариесрезистентность эмали. Последнюю в свою очередь определяют микротвердость, кислотоустойчивость и проницаемость. В понимании комплексной заботы о здоровье человека, и прежде всего детей, на первый план выступает превентивная точка зрения. Учитывая результаты «поперечного» эпидемиологического исследования на распространенность и интенсивность кариеса зубов у детского населения, прямое и непосредственное влияние оказывают, во-первых, экологические факторы, например, низкое содержание фторида в питьевой воде, загрязнение окружающей среды, неблагоприятные климатические условия. Во-вторых, социально-экономические, такие как ограниченная доступность стоматологической помощи, низкий уровень жизни и образования, характер питания, недостаточная заинтересованность в вопросах сохранения здоровья полости рта. Низкий уровень мотивации, санитарно-гигиенических знаний детского населения к сохранению стоматологического здоровья является фактором высокого риска развития патологического процесса твердых тканей, молочных и постоянных зубов детей. Резюмируя вышеизложенное, считается актуальным систематизация факторов риска возникновения кариеса для современного стоматологического просвещения детского населения, родителей и педагогов с упором на этиопатогенез Caries is one of the most urgent problems of modern dentistry and ranks first in frequency and intensity among all dental diseases. It is established that dental caries is the final stage of interaction of many endogenous and exogenous factors, individual for each patient. Most research studies on the causes of dental caries prove that the direct etiological factor is a local change in pH under the biofilm fixed on the tooth surface, due to the breakdown of carbohydrates by microorganisms and the formation of organic acids. It should be mentioned that the case trefoil serves as a graphic model of the theory of caries occurrence in modern dentistry, in which three conditions are equivalent, namely cariesogenic flora, easily fermentable carbohydrates, and low caries resistance of enamel. The latter, in turn, is determined by microhardness, acid resistance, and permeability. In the understanding of comprehensive care for human health, and especially for children, the preventive point of view comes to the fore. Taking into account the results of the "cross-section" epidemiological study, the prevalence and intensity of dental caries in the child population are directly and directly influenced, first, by environmental factors, such as low fluoride content in drinking water, environmental pollution, and unfavorable climatic conditions. Second, socio-economic factors, such as limited access to dental care, low standard of living and education, nutritional status, and lack of interest in maintaining oral health. The low level of motivation, sanitary and hygienic knowledge of the children's population to maintain dental health is a high risk factor for the development of the pathological process of hard tissues and milk, and permanent teeth of children. Summarizing the above, it is considered relevant to systematize the risk factors of caries for modern dental education of children's population, parents and teachers with an emphasis on etiopathogenesis

A new study has identified a vital gene in the pathogenesis and progression of liver cancer hepatocellular carcinoma (HCC), according to a team of biotechnology researchers at The American University in Cairo, Egypt, in a scientific paper published recently by AMOR. <br /><br /> The study on human gene ‘Cofactor of BRCA1’ (dubbed COBRA1) and its potential role as a reliable cancer predictor for HCC is especially important due to the disease’s grim outlook. HCC is “ranked as the second most common cause of cancer-related deaths in the world in 2012,” the authors said. “Thus, it is considered as a highly aggressive cancer with poor prognosis,” they added. <br /><br /> According to data from the Surveillance Epidemiology and End Results (SEER) program, hepatocellular carcinoma accounts for 90% of all liver cancers worldwide. In the United States, HCC represents the fastest growing cause of cancer mortality overall and the second fastest growing cause of cancer deaths among women. Globally, the incidence of HCC in developing nations is over twice that of in developed countries – East Asia having highest incidence of HCC with the rate of 35 male cases per 100,000, followed by the continent of Africa. <br /><br /> HCC mortality statistics in the developing countries is also more than double compared to the First World nations, with the annual loss of 33.5 and 23.73 lives per 100,000 in Asia and Africa, respectively. <br /><br /> In addition, “HCC is usually diagnosed in the late stages of the tumor where, at some point, treatment is of limited efficacy. Thus, prognoses and follow-ups are necessary to regularly assess the patients and to predict any risks before the deterioration of patients’ condition,” said researcher Aya Youssef and her fellow team members. <br /><br /><center><img src="http://www.piscomed.com/wp-content/uploads/2016/08/cobra1-2-quote.jpg" alt="" /></center><br /><br /> The behaviour of COBRA1 in the development and progression of several cancers has previously been studied and established, the researchers wrote. “For example, cell lines and tissues isolated from late-stage metastatic breast cancer tumors showed low expression levels of the COBRA1 protein, which has displayed tumor suppressor activity,” they noted, adding, “another study reported that COBRA1 was overexpressed in upper gastrointestinal carcinoma (UGC) tissue samples.” <br /><br /> Yet, COBRA1’s involvement in HCC tumor formation and growth has been subjected to minimal studies. “To date, the molecular mechanisms underlying HCC pathogenesis have not been fully identified, thus resulting in a lack of reliable prognostic markers for HCC,” said the researchers. <br /><br /> In their study, the Egyptian team hypothesized that the transcription factor COBRA1 is involved in HCC pathogenesis. The overexpression of COBRA1 is typically accompanied by the decreased expression of a tumor suppressor called trefoil factor-1 (TFF1). “COBRA1 controls TFF1 expression by regulating the activator protein-1 (AP-1) complex trans-activation, leading to attenuated TFF1 expression,” the researchers explained. <br /><br /> AP-1 regulates the invasive response in some tumors, hence paying a part in the mechanism of the spread of cancer. In addition, according to the researchers, “AP-1…consists of both Fos and Jun genes’ family members, which include c-Fos and c-Jun.” The c-Fos protein has been previously found to briefly up-regulated in HCC’s early stages and then quickly declines in the later stages of tumor progression – a process “for priming hepatocytes [predominant main cells of the liver] for migration and tissue invasion,” Youssef and her team explained. <br /><br /> In contrast, the c-Jun protein is involved in tumor cell survival and apoptosis by subduing cell death regulator p53, the widely recognized and important protein known as the “guardian of the genome” that plays a crucial role in thwarting genome mutation, and consequently preventing cancer. <br /><br /><center><img src="http://www.piscomed.com/wp-content/uploads/2016/08/cobra1-1-quote.jpg" alt="" /></center><br /><br /> Therefore, the researchers sought to investigate the expression of COBRA1, TFF1 and the AP-1 complex in tumour samples ranging from low-grade to high-grade HCC cell lines from patients at various stages of the disease. <br /><br /> “To the best of our knowledge, this is the first study to evaluate the mRNA expression levels of COBRA1 and its protein in cell lines representing advanced stages of HCC,” researchers said. <br /><br /> Their results revealed that the expressions of COBRA1, c-Fos, and c-Jun showed maximum levels in low-grade HCC cell line, i.e. tissues at early stages of cancer, while minimum levels of the three proteins were observed in high-grade HCC cell line representing late-stage tumor. <br /><br /> “Therefore, we suggest that COBRA1 displays tumor suppressor activity and may have a potential role in HCC progression,” they said. “We propose that these molecules could be used in the characterization of HCC cell lines at the molecular level.” <br /><br /> In addition, the researchers also examined COBRA1 levels to study HCC’s growth and spread. “In order to investigate the possible association of COBRA1 levels with HCC cell migration, we carried out the scratch wound healing assay to mimic the in vivo migratory behavior of HCC cells,” they described. <br /><br /> The result confirmed their theory. “The migratory ability of SNU-449 [late-stage cancer cell lines] that had relatively lower COBRA1 levels was significantly higher than that of the HepG2 [early-stage] cell lines with high COBRA1 expression,” observed the researchers, “which suggests a possible correlation between the low protein level of COBRA1 and tumor cell migration and thus, poor prognosis.” <br /><br /> While the researchers acknowledged that further studies are needed to examine the link between COBRA1, the AP-1 complex and TFF1 in HCC in order to draw a clearer conclusion, their study nevertheless suggest potential involvement of COBRA1 in the pathogenesis and development of HCC, as well as its function as tumor suppressor. <br /><br /> The study is co-authored by Aya Youssef, Heba Shawer, Alaa Afify and Asma Amleh, and is available online on AMOR.

ABSTRACT: A pot experiment was conducted to study the nodulating capacity of some forage legumes. Birdsfoot trefoil (Lotus corniculatus L), sainfoin (Onobrychis viciifolia Scop.) and subterranean clover (Trifolium subterraneum L.) were grown in pure culture (100%) and in mixtures with tall fescue (Festuca arundinacea Schreb.) in the ratios: birdsfoot trefoil + tall fescue (50:50%); sainfoin + tall fescue (50:50%); subterranean clover + tall fescue (50:50%); birdsfoot trefoil + subterranean clover + tall fescue (33:33:33%); sainfoin + subterranean clover + tall fescue (33:33:33%). Birdsfoot trefoil from the crops cultivated in pure stands showed the highest nodulation rating (31), followed by sainfoin (26) and subterranean clover (26). Nodulation rating in binary mixtures with grass component increased to 16.1% for birdsfoot trefoil and to 23.1% for subclover. Birdsfoot trefoil, both in pure stands and in mixtures, showed the highest phosphorus use efficiency for nodule formation.

Abstract TFF3 has been identified as a novel biomarker to distinguish between lung adenocarcinoma (ADC) and lung squamous-cell carcinoma (SCC). Herein, we determined the oncogenic functions of TFF3 and demonstrated the potential of pharmacological inhibition of TFF3 in lung ADC using a novel small-molecule inhibitor of TFF3 dimerization (AMPC). Forced expression of TFF3 in lung ADC cells enhanced cell proliferation and survival, increased anchorage-independent growth, cancer stem cell behavior, growth in 3D Matrigel, and cell migration and invasion. In contrast, depleted expression of TFF3 suppressed these cellular functions. Mechanistically, TFF3 exerted its oncogenic function through upregulation of ARAF and hence enhanced downstream activation of MEK1/2 and ERK1/2. Pharmacological inhibition of TFF3 by AMPC, resulted in markedly decreased cell survival, proliferation, 3D growth and foci formation, and impaired tumor growth in a xenograft mouse model. Moreover, the combination of various MEK1/2 inhibitors with AMPC exhibited synergistic inhibitory effects on lung ADC cell growth. In conclusion, this study provides the first evidence that TFF3 is a potent promoter of lung ADC progression. Targeting TFF3 with a novel small-molecule inhibitor alone or in combination with conventional MEK1/2 inhibitors are potential strategies to improve the outcome of lung ADC.

Purpose The purpose of this study is to determine the possibilities to use drill cuttings in soil formation processes on sandy substrates. The ecological and toxicological assessment of drill cuttings of various genesis and mixtures based on them is applied for the purpose. Design/methodology/approach Acute toxicity of mixtures consisting of various drill cuttings, sand and peat was estimated using soft wheat seeds (Triticum aestivum) using the eluate method. Subacute toxicity experiments were carried out using creeping trefoil (white clover) seeds (Trifolium repens), rye seeds (Secale cereale), and garden radish seeds (Raphanus sativus L.). Drill cuttings of the West Siberian oil-and-gas basin generated as a result of drilling on clay-polymer drilling fluids can be used as a component of soil-like mixtures in the reclamation of sand fills. Patterns of the selective stimulation of seed growth by components of drilling fluids (xanthate and bentonite) were revealed. Findings It was found that the addition of bentonite and xanthan (0.05% by weight of the cuttings each) reduces the suppression of seed growth occasioned salt content by 21.1% and 24.0%, respectively. Originality/value Soil degradation and desertification is a serious and widespread problem. The restoration of the fertile layer can be launched by application of the artificial soil-like mixtures based on drill cuttings of a certain origin to the disturbed lands.

Knots have attracted scientists in mathematics, physics, biology, and engineering. Long flexible thin strings easily knot and tangle as experienced in our daily life. Similarly, long polymer chains inevitably tend to get trapped into knots. Little is known about their formation or function in proteins despite &gt;1,000 knotted proteins identified in nature. However, these protein knots are not mathematical knots with their backbone polypeptide chains because of their open termini, and the presence of a “knot” depends on the algorithm used to create path closure. Furthermore, it is generally not possible to control the topology of the unfolded states of proteins, therefore making it challenging to characterize functional and physicochemical properties of knotting in any polymer. Covalently linking the amino and carboxyl termini of the deeply trefoil-knotted YibK from Pseudomonas aeruginosa allowed us to create the truly backbone knotted protein by enzymatic peptide ligation. Moreover, we produced and investigated backbone cyclized YibK without any knotted structure. Thus, we could directly probe the effect of the backbone knot and the decrease in conformational entropy on protein folding. The backbone cyclization did not perturb the native structure and its cofactor binding affinity, but it substantially increased the thermal stability and reduced the aggregation propensity. The enhanced stability of a backbone knotted YibK could be mainly originated from an increased ruggedness of its free energy landscape and the destabilization of the denatured state by backbone cyclization with little contribution from a knot structure. Despite the heterogeneity in the side-chain compositions, the chemically unfolded cyclized YibK exhibited several macroscopic physico-chemical attributes that agree with theoretical predictions derived from polymer physics.

Apolipoprotein (apo)A-I, the major protein constituent of high density lipoproteins (HDL), acts as a conformationally dynamic scaffold on the surface of the particle. Evidence suggests apoA-I conformation dictates HDL functionality and capacity to interface with other HDL proteins; however, the details of apoA-I structure and HDL formation are not completely understood. X-ray crystal structures of truncated forms of apoA-I and its cousin, apoA-IV, show reciprocal domain swapping structures in the lipid-free state. Recently, we showed the truncated mutant of apoA-I is also highly dynamic in solution with a flexible N-terminus. The purpose of this study was to a) derive a structural model of wild-type (WT) apoA-I using the new in-solution truncation model as a template and b) determine the relationship between monomeric, and oligomeric forms of lipid-free WT apoA-I. Using a novel isotope labeling strategy and chemical cross-linking, we defined spatial relationships within and between different molecules of WT apoA-I. The cross-linking patterns revealed a high degree of structural similarity between WT apoA-I oligomers. We found that monomeric and dimeric WT apoA-I structures were similar, but not identical, to those predicted by the crystal and solution structures of truncated apoA-I. Using cross-linking data, we derived two interconverting structures each for monomeric WT apoA-I, both containing dynamic N- and C-terminal regions. Similar structures were generated for WT dimeric and trimeric species. Small angle X-ray scattering was used to derive molecular envelopes of the molecules and refine the arrangements to propose the most detailed molecular models of lipid-free WT apoA-I to date. The models were used in conjunction with the reported double-belt and trefoil structures reconstituted HDL to reveal evidence of a parsimonious relationship between lipid-free and lipid-bound WT apoA-I. We propose a structural framework of WT apoA-I self-association and lipidation, centric to the C-terminal region of the molecule, that forms a basis for understanding HDL particle formation at the cell surface.

BackgroundRecent studies demonstrated that Aquamin®, a calcium-, magnesium-rich, multi-mineral natural product, improves barrier structure and function in colonoids obtained from the tissue of healthy subjects. The goal of the present study was to determine if the colonic barrier could be improved in tissue from subjects with ulcerative colitis (UC).MethodsColonoid cultures were established with colon biopsies from 9 individuals with UC. The colonoids were then incubated for a 2-week period under control conditions (in culture medium with a final calcium concentration of 0.25 mM) or in the same medium supplemented with Aquamin® to provide 1.5 – 4.5 mM calcium. Effects on differentiation and barrier protein expression were determined using several approaches: phase-contrast and scanning electron microscopy, quantitative histology and immunohistology, mass spectrometry-based proteome assessment and transmission electron microscopy.ResultsAlthough there were no gross changes in colonoid appearance, there was an increase in lumen diameter and wall thickness on histology and greater expression of cytokeratin 20 (CK20) along with reduced expression of Ki67 by quantitative immunohistology observed with intervention. In parallel, upregulation of several differentiation-related proteins was seen in a proteomic screen with the intervention. Aquamin®-treated colonoids demonstrated a modest up-regulation of tight junctional proteins but stronger induction of adherens junction and desmosomal proteins. Increased desmosomes were seen at the ultrastructural level. Proteomic analysis demonstrated increased expression of several basement membrane proteins and hemidesmosomal components. Proteins expressed at the apical surface (mucins and trefoils) were also increased as were several additional proteins with anti-microbial activity or that modulate inflammation. Finally, several transporter proteins that affect electrolyte balance (and, thereby affect water resorption) were increased. At the same time, growth and cell cycle regulatory proteins (Ki67, nucleophosmin, and stathmin) were significantly down-regulated. Laminin interactions, matrix formation and extracellular matrix organization were the top three up-regulated pathways with the intervention.ConclusionA majority of individuals including patients with UC do not reach the recommended daily intake for calcium and other minerals. To the extent that such deficiencies might contribute to the weakening of the colonic barrier, the findings employing UC tissue-derived colonoids here suggest that adequate mineral intake might improve the colonic barrier.