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El-Naggar, Sabry Ali, Karim Samy El-Said, Mona Elwan, Maysa Mobasher, Fotouh Mansour, Mohamed Elbakry, and Doaa Ibrahim Kabil. "Toxicity of bean cooking media containing EDTA in mice." Toxicology and Industrial Health 36, no. 6 (June 2020): 436–45. http://dx.doi.org/10.1177/0748233719893178.
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The possible renal and hepatic toxicities of ethylenediaminetetraacetic acid (EDTA) in bean cooking media were studied using 100 male albino mice. Two sublethal doses of EDTA were used to explore their toxic effects; 20 mg/kg and 200 mg/kg, which corresponded to 1/100th and 1/10th of LD50, respectively. Accordingly, the toxicity study was performed using 50 mice, divided into five groups ( n = 10/group) as follows: group 1 (Gp1) served as a negative control and was orally administered normal saline; group 2 (Gp2) was administered the bean cooking medium; group 3 (Gp3) was administered EDTA (200 mg/kg); group 4 (Gp4) was administered bean cooking medium containing 20 mg/kg of EDTA; and group 5 (Gp5) was administered bean cooking medium containing 200 mg/kg of EDTA. The results showed no significant changes in liver and kidney functions in Gp2 while Gp3, Gp4, and Gp5 exhibited significant increases in adverse liver and kidney function markers. Hematocrit values were significantly decreased in Gp3 and Gp5, while the total white blood cells counts were significantly decreased in Gp3 and significantly increased in Gp5. The number of platelets was decreased in Gp3, Gp4, and Gp5. The blood levels of sodium (Na+), iron (Fe2+), and calcium (Ca2+) were decreased in Gp3, Gp4, and Gp5 due to the chelating effects of EDTA. The hepatic and renal architectures were disorganized in Gp3, Gp4, and Gp5 with some hemorrhagic manifestations in livers and kidneys of mice. These results demonstrate that EDTA in bean cooking is harmful in mice under the conditions of this study, and the potentially harmful effects in humans supports restricting its use.
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Elfaky, Mahmoud A., Alaa Sirwi, Sameh H. Ismail, Heba H. Awad, and Sameh S. Gad. "Hepatoprotective Effect of Silver Nanoparticles at Two Different Particle Sizes: Comparative Study with and without Silymarin." Current Issues in Molecular Biology 44, no. 7 (June 30, 2022): 2923–38. http://dx.doi.org/10.3390/cimb44070202.
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Silver nanoparticles have been used for numerous therapeutic purposes because of their increased biodegradability and bioavailability, yet their toxicity remains questionable as they are known to interact easily with biological systems because of their small size. This study aimed to investigate and compare the effect of silver nanoparticles’ particle size in terms of their potential hazard, as well as their potential protective effect in an LPS-induced hepatotoxicity model. Liver slices were obtained from Sprague Dawley adult male rats, and the thickness of the slices was optimized to 150 μm. Under regulated physiological circumstances, freshly cut liver slices were divided into six different groups; GP1: normal, GP2: LPS (control), GP3: LPS + AgNpL (positive control), GP4: LPS + silymarin (standard treatment), GP5: LPS + AgNpS + silymarin (treatment I), GP6: LPS + AgNpL + silymarin (treatment II). After 24 h of incubation, the plates were gently removed, and the supernatant and tissue homogenate were all collected and then subjected to the following biochemical parameters: Cox2, NO, IL-6, and TNF-α. The LPS elicited marked hepatic tissue injury manifested by elevated cytokines and proinflammatory markers. Both small silver nanoparticles and large silver nanoparticles efficiently attenuated LPS hepatotoxicity, mainly via preserving the cytokines’ level and diminishing the inflammatory pathways. In conclusion, large silver nanoparticles exhibited effective hepatoprotective capabilities over small silver nanoparticles.
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Kaur, Rajinder, Sandeep Kaur, Neelima R. Kumar, and Kusum Harjai. "Honey bee collected pollen and beebread of Zea mays: determination of bioactive constituents and health benefits." Research Journal of Biotechnology 18, no. 3 (February 15, 2023): 64–69. http://dx.doi.org/10.25303/1803rjbt64069.
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Bee products are known since ancient times for their nutritional value and beneficial effects. The present study was carried out to evaluate the phytochemical composition and therapeutic potential of bee pollen and beebread. Broth dilution method was used to observe the in vitro antibacterial activity. For in vivo antioxidant activities, BALB/c mice were divided into six groups; Gp1 was given normal saline only, Gp2 was injected intraperitoneally with Salmonella enterica serovar Typhimurium at 2×104 CFU/ml, Gp3 was administrated orally with bee collected pollen of Zea mays (250mg/kg bw) only, Gp4 was treated with the bee collected pollen of Z. mays (250mg/kg bw) in Salmonella infected mice, Gp5 was administrated orally with beebread of Z. mays (250mg/kg bw) only and Gp6 was treated with the beebread of Z. mays (250mg/kg bw) in Salmonella infected mice. The results obtained in the study suggested that various bioactive constituents were present in bee pollen and beebread and these phytochemical constituents were responsible for the antibacterial and antioxidant efficacy of these studied bee products. Beebread was found to possess high activity as compared to bee pollen reflecting its high polphenolic composition. Hence, it could be inferred that bee pollen could be seen as a potential source for designing a drug against S. typhimurium.
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Kaisarly, Dalia, D. Meierhofer, M. El Gezawi, P. Rösch, and K. H. Kunzelmann. "Effects of flowable liners on the shrinkage vectors of bulk-fill composites." Clinical Oral Investigations 25, no. 8 (January 27, 2021): 4927–40. http://dx.doi.org/10.1007/s00784-021-03801-2.
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Abstract Objectives This investigation evaluated the effect of flowable liners beneath a composite restoration applied via different methods on the pattern of shrinkage vectors. Methods Forty molars were divided into five groups (n = 8), and cylindrical cavities were prepared and bonded with a self-etch adhesive (AdheSe). Tetric EvoCeram Bulk Fill (TBF) was used as the filling material in all cavities. The flowable liners Tetric EvoFlow Bulk Fill (TEF) and SDR were used to line the cavity floor. In gp1-TBF, the flowable composite was not used. TEF was applied in a thin layer in gp2-fl/TEF + TBF and gp3-fl/TEF + TBFincremental. Two flowable composites with a layer thickness of 2 mm were compared in gp4-fl/TEF + TBF and gp5-fl/SDR + TBF. TEF and SDR were mixed with radiolucent glass beads, while air bubbles inherently present in TBF served as markers. Each material application was scanned twice by micro-computed tomography before and after light curing. Scans were subjected to image segmentation for calculation of the shrinkage vectors. Results The absence of a flowable liner resulted in the greatest shrinkage vectors. A thin flowable liner (gp2-fl/TEF + TBFbulk) resulted in larger overall shrinkage vectors for the whole restoration than a thick flowable liner (gp4-fl/TEF + TBF). A thin flowable liner and incremental application (gp3-fl/TEF + TBFincremental) yielded the smallest shrinkage vectors. SDR yielded slightly smaller shrinkage vectors for the whole restoration than that observed in gp4-fl/TEF + TBF. Conclusions Thick flowable liner layers had a more pronounced stress-relieving effect than thin layers regardless of the flowable liner type. Clinical relevance It is recommended to apply a flowable liner (thin or thick) beneath bulk-fill composites, preferably incrementally.
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Zhang, Jianqiang, Peter J. Timoney, N. James MacLachlan, William H. McCollum, and Udeni B. R. Balasuriya. "Persistent Equine Arteritis Virus Infection in HeLa Cells." Journal of Virology 82, no. 17 (June 25, 2008): 8456–64. http://dx.doi.org/10.1128/jvi.01249-08.
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ABSTRACT The horse-adapted virulent Bucyrus (VB) strain of equine arteritis virus (EAV) established persistent infection in high-passage-number human cervix cells (HeLa-H cells; passages 170 to 221) but not in low-passage-number human cervix cells (HeLa-L cells; passages 95 to 115) or in several other cell lines that were evaluated. However, virus recovered from the 80th passage of the persistently infected HeLa-H cells (HeLa-H-EAVP80) readily established persistent infection in HeLa-L cells. Comparative sequence analysis of the entire genomes of the VB and HeLa-H-EAVP80 viruses identified 16 amino acid substitutions, including 4 in the replicase (nsp1, nsp2, nsp7, and nsp9) and 12 in the structural proteins (E, GP2, GP3, GP4, and GP5). Reverse genetic studies clearly showed that substitutions in the structural proteins but not the replicase were responsible for the establishment of persistent infection in HeLa-L cells by the HeLa-H-EAVP80 virus. It was further demonstrated that recombinant viruses with substitutions in the minor structural proteins E and GP2 or GP3 and GP4 were unable to establish persistent infection in HeLa-L cells but that recombinant viruses with combined substitutions in the E (Ser53→Cys and Val55→Ala), GP2 (Leu15→Ser, Trp31→Arg, Val87→Leu, and Ala112→Thr), GP3 (Ser115→Gly and Leu135→Pro), and GP4 (Tyr4→His and Ile109→Phe) proteins or with a single point mutation in the GP5 protein (Pro98→Leu) were able to establish persistent infection in HeLa-L cells. In summary, an in vitro model of EAV persistence in cell culture was established for the first time. This system can provide a valuable model for studying virus-host cell interactions, especially virus-receptor interactions.
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Go, Yun Young, Jianqiang Zhang, Peter J. Timoney, R. Frank Cook, David W. Horohov, and Udeni B. R. Balasuriya. "Complex Interactions between the Major and Minor Envelope Proteins of Equine Arteritis Virus Determine Its Tropism for Equine CD3+ T Lymphocytes and CD14+ Monocytes." Journal of Virology 84, no. 10 (March 10, 2010): 4898–911. http://dx.doi.org/10.1128/jvi.02743-09.
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ABSTRACT Extensive cell culture passage of the virulent Bucyrus (VB) strain of equine arteritis virus (EAV) to produce the modified live virus (MLV) vaccine strain has altered its tropism for equine CD3+ T lymphocytes and CD14+ monocytes. The VB strain primarily infects CD14+ monocytes and a small subpopulation of CD3+ T lymphocytes (predominantly CD4+ T lymphocytes), as determined by dual-color flow cytometry. In contrast, the MLV vaccine strain has a significantly reduced ability to infect CD14+ monocytes and has lost its capability to infect CD3+ T lymphocytes. Using a panel of five recombinant chimeric viruses, we demonstrated that interactions among the GP2, GP3, GP4, GP5, and M envelope proteins play a major role in determining the CD14+ monocyte tropism while the tropism for CD3+ T lymphocytes is determined by the GP2, GP4, GP5, and M envelope proteins but not the GP3 protein. The data clearly suggest that there are intricate interactions among these envelope proteins that affect the binding of EAV to different cell receptors on CD3+ T lymphocytes and CD14+ monocytes. This study shows, for the first time, that CD3+ T lymphocytes may play an important role in the pathogenesis of equine viral arteritis when horses are infected with the virulent strains of EAV.
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Morel-Kopp, Marie-Christine, Jeannine M. Clemetson, Kenneth J. Clemetson, Riitta Kekomaki, Hartmut Kroll, Katerina Michaelides, Edward G. D. Tuddenham, Karen Vanhoorelbeke, Christopher M. Ward, and Hai Po Helena Liang. "A common ancestral glycoprotein (GP) 9 1828A>G (Asn45Ser) gene mutation occurring in European families from Australia and Northern Europe with Bernard-Soulier syndrome (BSS)." Thrombosis and Haemostasis 94, no. 09 (2005): 599–605. http://dx.doi.org/10.1160/th05-03-0165.
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SummaryBernard-Soulier syndrome (BSS) is an extremely rare hereditary bleeding disorder, caused by mutations occurring in the Glycoprotein (GP) Ibα, GPIbβ and GP9 genes that encode for the corresponding subunits of platelet GPIb-V-IX adhesion receptor complex. BSS has been reported in many populations, mostly behaving in an autosomal-recessive manner. While the great majority of BSS mutations are unique to a single individual or family, the GP9 1828A>G Asn45Ser mutation, which we have identified in an undocumented Australian Caucasian, has already been reported in multiple unrelated Caucasian families from various Northern and Central European countries. Haplotype analysis of 19 BSS patients from 15 unrelated Northern European families (including 2 compound heterozygote siblings from a British family previously published, and 17 1828A>G Asn45Ser homozygotes), showed that 14 of these BSS patients from 11 of the 1828A>G Asn45Ser homozygote families share a common haplotype at the chromosomal region 3’ to the GP9 gene. Hence, the results suggest that the GP9 1828A>GAsn45Ser mutation in these families is ancient, and its frequent emergence in the European population is the result of a founder effect rather than recurrent mutational events. Association of the 1828A>G Asn45Ser mutation with variant haplotypes in 4 other Northern European BSS families raised the possibility of a second founder event, or rare recombinations in these families. Additional members from these ‘atypical’ lineages would need to be screened to resolve this question.
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Human, M.Sc, Prof Dr Soeranto, S. Loekito, M. Trilaksono, and A. Syaifudin. "Pemuliaan Mutasi Tanaman Nanas (Ananas comosus (L.) Merr.) Menggunakan Iradiasi Gamma untuk Perbaikan Varietas Nanas Smooth Cayenne." Jurnal Ilmiah Aplikasi Isotop dan Radiasi 12, no. 1 (January 31, 2017): 13. http://dx.doi.org/10.17146/jair.2016.12.1.3197.
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Pemuliaan Mutasi Tanaman Nanas (Ananas comosus (L.) Merr.) Menggunakan Iradiasi Gamma untuk Perbaikan Varietas Smooth Cayenne. Pada saat ini, jenis nanas yang paling banyak dibudidayakan dalam berbagai perdagangan dunia adalah jenis Smooth Cayenne. Banyak klon yang berasal dari kultivar ini seperti GP1, GP2, GP3, GP4, GP5, dan F180 ditanam oleh GGP baik untuk buahsegar dan olahan. GGPC mulai melakukan usaha perbaikan varietas nanas sejak tahun 1986, yaitu dengan tujuan meningkatkan kualitas dan produksi yang tinggi. Pemuliaan mutasi nanas dimulai pada tahun 2006, yaitu melalui kerjasama dengan Pusat Aplikasi Isotop dan Radiasi (PAIR), Badan Tenaga Nuklir Nasional (BATAN). Sejumlah 10 mahkota (crown) nanas berasal dari GP2, GP3 (A10) dan klon F180 diiradiasi sinar gamma bersumber Cobalt-60 terpasang pada iradiator gamma chamber 4000A dengan dosis 200 dan 300 Gy. Crown yang telah diradiasi kemudian ditanam di lahan percobaan dengan mengikuti budidaya standar komersial di PT. GGPC. Secara umum hasil penelitian menunjukkan tidak ada perbedaan yang signifikan antara kedua dosis iradiasi (200 dan 300 Gy) pada fenotipe tanaman. Namun, keragaman fenotipe yang sangat tinggi dijumpai pada klon turunan tanaman vegetatif (V1). Beberapa variasi tanaman yang muncul tercatat sebagai berikut: 47% tanaman normal, 15% Rosset, 11% berduri, 5% crown bercabang, 4% tanaman memiliki banyak daun dan 18% buah berbentuk abnormal. Variasi mutan yang signifikan juga diamati pada klon-lon turunan tanaman V2 dan beberapa mutan tampak lebih stabil pada generasi V3. Program pemuliaan nanas ini akan terus dilanjutkan untuk evaluasi yang berkaitan dengan perbaikan produktivitas, kualitas, dan ketahanan terhadap hama dan penyakit tertentu.
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Wieringa, Roeland, Antoine A. F. de Vries, Jannes van der Meulen, Gert-Jan Godeke, Jos J. M. Onderwater, Hans van Tol, Henk K. Koerten, A. Mieke Mommaas, Eric J. Snijder, and Peter J. M. Rottier. "Structural Protein Requirements in Equine Arteritis Virus Assembly." Journal of Virology 78, no. 23 (December 1, 2004): 13019–27. http://dx.doi.org/10.1128/jvi.78.23.13019-13027.2004.
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ABSTRACT Equine arteritis virus (EAV) is an enveloped, positive-stranded RNA virus belonging to the family Arteriviridae of the order Nidovirales. EAV particles contain seven structural proteins: the nucleocapsid protein N, the unglycosylated envelope proteins M and E, and the N-glycosylated membrane proteins GP2b (previously named GS), GP3, GP4, and GP5 (previously named GL). Proteins N, M, and GP5 are major virion components, E occurs in virus particles in intermediate amounts, and GP4, GP3, and GP2b are minor structural proteins. The M and GP5 proteins occur in virus particles as disulfide-linked heterodimers while the GP4, GP3, and GP2b proteins are incorporated into virions as a heterotrimeric complex. Here, we studied the effect on virus assembly of inactivating the structural protein genes one by one in the context of a (full-length) EAV cDNA clone. It appeared that the three major structural proteins are essential for particle formation, while the other four virion proteins are dispensable. When one of the GP2b, GP3, or GP4 proteins was missing, the incorporation of the remaining two minor envelope glycoproteins was completely blocked while that of the E protein was greatly reduced. The absence of E entirely prevented the incorporation of the GP2b, GP3, and GP4 proteins into viral particles. EAV particles lacking GP2b, GP3, GP4, and E did not markedly differ from wild-type virions in buoyant density, major structural protein composition, electron microscopic appearance, and genomic RNA content. On the basis of these results, we propose a model for the EAV particle in which the GP2b/GP3/GP4 heterotrimers are positioned, in association with a defined number of E molecules, above the vertices of the putatively icosahedral nucleocapsid.
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Olson, N. H., W. Xu, W. D. Grochulski, D. L. Andersont, and T. S. Baker. "Electron Microscopy of Negatively Stained and Frozen-Hydrated Bacteriophage Φ29." Proceedings, annual meeting, Electron Microscopy Society of America 48, no. 1 (August 12, 1990): 270–71. http://dx.doi.org/10.1017/s0424820100180100.
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Bacteriophage Φ29 is a complex-shaped, double-stranded DNA virus that infects Bacillus subtilis . Mature Φ29 particles contain six major structural proteins which give rise to several distinctive features: a prolate-shaped head (gp8), fibers (gp8.5) which radiate from the head, a connector-collar region (gp10 and gp11), collar appendages (gp12*) and a tail (gp9). Although much is known about Φ29 biochemistry, genetics and assembly, relatively little is known about its detailed structure. Three-dimensional reconstructions of the collar demonstrated that it contains two structurally distinct features: an upper portion (proximal to the head) with 12-fold axial symmetry (gp10) and a lower protion with 6-fold symmetry (possibly both gp10 and gp11). The capsid is believed to be organized with two apical regions with trimeric aggregates of gp8 dimers arranged on a T=l icosahedral lattice separated by 10 additional trimers in the equatorial region. Φ29 proheads are assembly intermediates that contain only gp8, gp8.5, gp10 and the scaffolding protein (gp7) which is released upon DNA encapsidation. We have imaged Φ29 with conventional negative-staining and the recently developed cryo-microscopy techniques with the goal of examining the structural basis for Φ29 morphogenesis.Vitrified Φ29 proheads display distinct outlines with one end pointed and the other somewhat flattened and they often orient with their long axes in the plane of the vitreous water (Fig. 1). Occasional end-on views with a circular profile are also observed. Proheads show a tendency to associate in pairs with the closest contact between the flattened ends (Fig. 1, arrow-head). Longer morphological variants are also occassionally observed (Fig. 1, arrow). The morphology of proheads is dramatically distorted in negative-stain preparations indicating significantly poorer preservation using this technique (Fig. 2). Proheads are also permeable to stain.
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Matczuk, Anna Karolina, Minze Zhang, Michael Veit, and Maciej Ugorski. "Expression of the Heterotrimeric GP2/GP3/GP4 Spike of an Arterivirus in Mammalian Cells." Viruses 14, no. 4 (April 1, 2022): 749. http://dx.doi.org/10.3390/v14040749.
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Equine arteritis virus (EAV), an enveloped positive-strand RNA virus, is an important pathogen of horses and the prototype member of the Arteiviridae family. Unlike many other enveloped viruses, which possess homotrimeric spikes, the spike responsible for cellular tropism in Arteriviruses is a heterotrimer composed of 3 glycoproteins: GP2, GP3, and GP4. Together with the hydrophobic protein E they are the minor components of virus particles. We describe the expression of all 3 minor glycoproteins, each equipped with a different tag, from a multi-cassette system in mammalian BHK-21 cells. Coprecipitation studies suggest that a rather small faction of GP2, GP3, and GP4 form dimeric or trimeric complexes. GP2, GP3, and GP4 co-localize with each other and also, albeit weaker, with the E-protein. The co-localization of GP3-HA and GP2-myc was tested with markers for ER, ERGIC, and cis-Golgi. The co-localization of GP3-HA was the same regardless of whether it was expressed alone or as a complex, whereas the transport of GP2-myc to cis-Golgi was higher when this protein was expressed as a complex. The glycosylation pattern was also independent of whether the proteins were expressed alone or together. The recombinant spike might be a tool for basic research but might also be used as a subunit vaccine for horses.
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Wunderlich, Frank, Denis Delic, Daniela Gerovska, and Marcos J. Araúzo-Bravo. "Vaccination Accelerates Liver-Intrinsic Expression of Megakaryocyte-Related Genes in Response to Blood-Stage Malaria." Vaccines 10, no. 2 (February 14, 2022): 287. http://dx.doi.org/10.3390/vaccines10020287.
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Erythropoiesis and megakaryo-/thrombopoiesis occur in the bone marrow proceeding from common, even bipotent, progenitor cells. Recently, we have shown that protective vaccination accelerates extramedullary hepatic erythroblastosis in response to blood-stage malaria of Plasmodium chabaudi. Here, we investigated whether protective vaccination also accelerates extramedullary hepatic megakaryo-/thrombopoiesis. Female Balb/c mice were twice vaccinated with a non-infectious vaccine before infecting with 106 P. chabaudi-parasitized erythrocytes. Using gene expression microarrays and quantitative real-time PCR, transcripts of genes known to be expressed in the bone marrow by cells of the megakaryo-/thrombocytic lineage were compared in livers of vaccination-protected and unprotected mice on days 0, 1, 4, 8, and 11 p.i. Livers of vaccination-protected mice responded with expression of megakaryo-/thrombocytic genes faster to P. chabaudi than those of unvaccinated mice, evidenced at early patency on day 4 p.i., when livers exhibited significantly higher levels of malaria-induced transcripts of the genes Selp and Pdgfb (p-values < 0.0001), Gp5 (p-value < 0.001), and Fli1, Runx1, Myb, Mpl, Gp1ba, Gp1bb, Gp6, Gp9, Pf4, and Clec1b (p-values < 0.01). Together with additionally analyzed genes known to be related to megakaryopoiesis, our data suggest that protective vaccination accelerates liver-intrinsic megakaryo-/thrombopoiesis in response to blood-stage malaria that presumably contributes to vaccination-induced survival of otherwise lethal blood-stage malaria.
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Mobasher, Maysa A., Mousa O. Germoush, Hala Galal El-Tantawi, and Karim Samy El-Said. "Metformin Improves Biochemical and Pathophysiological Changes in Hepatocellular Carcinoma with Pre-Existed Diabetes Mellitus Rats." Pathogens 10, no. 1 (January 11, 2021): 59. http://dx.doi.org/10.3390/pathogens10010059.
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Hepatocellular carcinoma (HCC) is one of the world’s most widely recognized malignant tumors that accounts for 90% of all the primary liver cancers and is a major cause of death from cancer, representing half a million deaths per year. Obesity and associated metabolic irregularities, particularly diabetes mellitus (DM) and insulin resistance, are important risk factors for the advancement of HCC. Recently, retrospective studies showed that metformin (MET) could protect the hepatic tissues in pre-existing diabetes mellitus from HCC. The purpose of this study was to assess the role of MET treatment in the pre-existing diabetic rats before and after HCC induction by diethylnitrosamine (DEN). Thirty-five male Sprague Dawley albino rats were partitioned into the following groups: Group 1 (Gp1) was the control. Gp2 was injected intraperitoneally (i.p) with streptozotocin (STZ) (80 mg/kg) and DEN (50 mg/kg/7 weeks). Gp3, Gp4, and Gp5 were injected as in Gp2 and treated with MET (150 mg/kg) before and/or after HCC induction. Biochemical parameters including liver functions, lipid profile, and oxidative stress biomarkers were determined. Furthermore, histological and immunohistochemical changes were assessed in all groups. Our results illustrated that the group of rats that were treated with STZ and DEN had significant changes in both liver functions and were associated with alterations in the liver histopathological architectures. Treatment with MET before or after HCC induction ameliorated the cellular changes in the liver tissues; however, the utmost protection was found in a group of rats, which were treated with MET before and after HCC induction.
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Mobasher, Maysa A., Mousa O. Germoush, Hala Galal El-Tantawi, and Karim Samy El-Said. "Metformin Improves Biochemical and Pathophysiological Changes in Hepatocellular Carcinoma with Pre-Existed Diabetes Mellitus Rats." Pathogens 10, no. 1 (January 11, 2021): 59. http://dx.doi.org/10.3390/pathogens10010059.
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Hepatocellular carcinoma (HCC) is one of the world’s most widely recognized malignant tumors that accounts for 90% of all the primary liver cancers and is a major cause of death from cancer, representing half a million deaths per year. Obesity and associated metabolic irregularities, particularly diabetes mellitus (DM) and insulin resistance, are important risk factors for the advancement of HCC. Recently, retrospective studies showed that metformin (MET) could protect the hepatic tissues in pre-existing diabetes mellitus from HCC. The purpose of this study was to assess the role of MET treatment in the pre-existing diabetic rats before and after HCC induction by diethylnitrosamine (DEN). Thirty-five male Sprague Dawley albino rats were partitioned into the following groups: Group 1 (Gp1) was the control. Gp2 was injected intraperitoneally (i.p) with streptozotocin (STZ) (80 mg/kg) and DEN (50 mg/kg/7 weeks). Gp3, Gp4, and Gp5 were injected as in Gp2 and treated with MET (150 mg/kg) before and/or after HCC induction. Biochemical parameters including liver functions, lipid profile, and oxidative stress biomarkers were determined. Furthermore, histological and immunohistochemical changes were assessed in all groups. Our results illustrated that the group of rats that were treated with STZ and DEN had significant changes in both liver functions and were associated with alterations in the liver histopathological architectures. Treatment with MET before or after HCC induction ameliorated the cellular changes in the liver tissues; however, the utmost protection was found in a group of rats, which were treated with MET before and after HCC induction.
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Y.E, El-Bolkiny. "Treatment with Rosemarinus Officinalis Leaves and Saussurea Lappa Roots Extracts Ameliorate Hepatorenal and Cardiac Cisplatin Toxicity in Ehrlich Ascitic-Bearing Mice." Clinical Oncology Case Reports 1, no. 1 (September 23, 2022): 01–04. http://dx.doi.org/10.31579/2834-5061/003.
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Medicinal plants have important role in cancer treatments and protect the vital organs from the adverse effect of chemotherapy treatments. This study was conducted to investigate the ameliorating effects of Rosemarinus officinalis leaves (ROLE) and Saussurea lappa roots (SLRE) extracts along with cisplatin (Cis) on hepato-renal and cardiac-toxicities induced in Ehrlich-bearing mice. Forty female Swiss albino mice were divided into 4 groups (n=10) as follows; the first group (Gp1) was served as control. Gp2-Gp4 were inoculated with 1 x 106 EAC-cells/mouse. Gp2 was left without any treatment as EAC-bearing mice, and Gp3 had injected with Cis (2 mg/Kg) starting from day 3 to day7. Gp4 had injected with Cis as in Gp3, and then administered with ROLE/ SLRE from day 3 to day 13. At day 14, blood samples were collected and heart tissue homogenate was prepared for biochemical analysis. Serum hepatic enzymes; alanine aminotransferase (ALT), aspartate aminotransferase (AST) ɤ-glutamyltransferase (ɤ GGT), bilirubin, creatinine and urea levels were significantly increased in EAC-bearing mice, however, the treatment of EAC-bearing mice with Cis/ROLE/SLRE decreased the mentioned biochemical parameters. The contents of troponin (TnI) and creatine-phosphokinase (CPK) in the heart tissue of EAC-bearing mice were also increased; however, the treatment with Cis/ROLE/SLRE exhibited a marked ameliorative effect on both Tn1 and CPK. These results suggested that administration of ROLE/SLRE along with Cis mitigated the toxicity in EAC-bearing mice
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Cole, S. R., G. Young, D. Byrne, J. Guy, and J. Morcom. "Participation in screening for colorectal cancer based on a faecal occult blood test is improved by endorsement by the primary care practitioner." Journal of Medical Screening 9, no. 4 (December 1, 2002): 147–52. http://dx.doi.org/10.1136/jms.9.4.147.
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OBJECTIVES: To investigate the influence of general practitioner (GP) endorsement on participation in screening for colorectal cancer based on a faecal occult blood test (FOBT). SETTING: South Australian residents (n=2400), in 1999, aged >50 years. METHODS: Random selection of three groups (GP1, GP2, GP3) from two general practices and of one group (ER) from the federal electoral roll; n=600 per group. Without previous communication or publicity, subjects were posted an offer of screening by immunochemical FOBT. The GP1 and ER groups were invited without indication that their GP was involved; GP2 received an invitation indicating support from the practice; and GP3 received an invitation on practice letterhead and signed by a practice partner. A reminder was posted at 6 weeks. Participation was defined as return of correctly completed FOBT sample cards within 12 weeks. RESULTS: Participation rates were: GP1 192/600 (32.0%), GP2 228/600 (38.0%), and GP3 244/600 (40.7%); &khgr;2=10.2, p=0.006. Both GP2 and GP3 differed significantly from GP1 (odds ratio (OR) 0.77, 95% confidence interval (95% CI) 0.60 to 0.98 and relative risk (RR)=0.69, 95% CI 0.54 to 0.87 respectively). ER (193/600 (32.2%)) and GP1 were not significantly different. Age but not sex was significantly associated with participation. Overall test positivity rate was 4.6%; five malignancies were found in the 918 who performed FOBT. CONCLUSIONS: Association of a GP of recent contact with a screening offer in the form of a personalised letter of invitation achieves better participation than does the same letter from a centralised screening unit that does not mention the GP. Thus, GP enhanced participation is achievable without their actual involvement. Additional strategies are needed to further improve participation.
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ABIODUN ALONGE, SEGUN, and CHRISTOPHER OSITA ANYAECHE. "DEVELOPMENT OF A MULTI-OBJECTIVE PRODUCTION PLANNING MODEL FOR A SAWMILL." Journal of Engineering Studies and Research 27, no. 1 (June 7, 2021): 21–32. http://dx.doi.org/10.29081/jesr.v27i1.248.
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Based on the objectives set out for a Sawmill, a goal programming model was developed to simultaneously consider the production volumes goal, sales revenue goal, production cost goal, and machine utilization goal in order to develop its production plans for a horizon. The unwanted deviations from the goals served as the objective function to be optimized subject to the goals constraints, operational constraints, and non-negativity constraints. Three independent pre-emptive goal priority structures, GP1, GP2, and GP3, were considered with each prioritizing the objectives differently. The goal programming model was tested for its utility using data gathered from the mill to the three-goal priority structures. The results obtained indicated that, for GP1, the product volume goals for all products were achieved, and all but one, volume goals were achieved for both GP2 and GP3. The viability test showed that all priority structures used were profitable with GP1, GP2, and GP3 recording 1.099, 1.102, and 1.095 respectively. This indicates that the three priority structures considered are approximately profitable at the same level. The goal programming model for production planning offers the decision-maker a variety of options as to its application.
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ABIODUN ALONGE, SEGUN, and CHRISTOPHER OSITA ANYAECHE. "DEVELOPMENT OF A MULTI-OBJECTIVE PRODUCTION PLANNING MODEL FOR A SAWMILL." Journal of Engineering Studies and Research 27, no. 1 (June 7, 2021): 21–32. http://dx.doi.org/10.29081/jesr.v27i1.248.
Full textAbstract:
Based on the objectives set out for a Sawmill, a goal programming model was developed to simultaneously consider the production volumes goal, sales revenue goal, production cost goal, and machine utilization goal in order to develop its production plans for a horizon. The unwanted deviations from the goals served as the objective function to be optimized subject to the goals constraints, operational constraints, and non-negativity constraints. Three independent pre-emptive goal priority structures, GP1, GP2, and GP3, were considered with each prioritizing the objectives differently. The goal programming model was tested for its utility using data gathered from the mill to the three-goal priority structures. The results obtained indicated that, for GP1, the product volume goals for all products were achieved, and all but one, volume goals were achieved for both GP2 and GP3. The viability test showed that all priority structures used were profitable with GP1, GP2, and GP3 recording 1.099, 1.102, and 1.095 respectively. This indicates that the three priority structures considered are approximately profitable at the same level. The goal programming model for production planning offers the decision-maker a variety of options as to its application.
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Zheng, Jing, Wenyuan Chen, Hao Xiao, Fan Yang, Xiaowu Li, Jingdong Song, Lingpeng Cheng, and Hongrong Liu. "A Capsid Structure of Ralstonia solanacearum podoviridae GP4 with a Triangulation Number T = 9." Viruses 14, no. 11 (November 1, 2022): 2431. http://dx.doi.org/10.3390/v14112431.
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GP4, a new Ralstonia solanacearum phage, is a short-tailed phage. Few structures of Ralstonia solanacearum phages have been resolved to near-atomic resolution until now. Here, we present a 3.7 Å resolution structure of the GP4 head by cryo-electron microscopy (cryo-EM). The GP4 head contains 540 copies of major capsid protein (MCP) gp2 and 540 copies of cement protein (CP) gp1 arranged in an icosahedral shell with a triangulation number T = 9. The structures of gp2 and gp1 show a canonical HK97-like fold and an Ig-like fold, respectively. The trimeric CPs stick on the surface of the head along the quasi-threefold axis of the icosahedron generating a sandwiched three-layer electrostatic complementary potential, thereby enhancing the head stability. The assembly pattern of the GP4 head provides a platform for the further exploration of the interaction between Ralstonia solanacearum and corresponding phages.
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Wieringa, Roeland, Antoine A. F. de Vries, and Peter J. M. Rottier. "Formation of Disulfide-Linked Complexes between the Three Minor Envelope Glycoproteins (GP2b, GP3, and GP4) of Equine Arteritis Virus." Journal of Virology 77, no. 11 (June 1, 2003): 6216–26. http://dx.doi.org/10.1128/jvi.77.11.6216-6226.2003.
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ABSTRACT Equine arteritis virus (EAV) is an enveloped, positive-stranded RNA virus belonging to the family Arteriviridae of the order Nidovirales. Six transmembrane proteins have been identified in EAV particles: the nonglycosylated membrane protein M and the glycoprotein GP5 (previously named GL), which occur as disulfide-bonded heterodimers and are the major viral envelope proteins; the unglycosylated small envelope protein E; and the minor glycoproteins GP2b (formerly designated GS), GP3, and GP4. Analysis of the appearance of the GP2b, GP3, and GP4 proteins in viral particles by gel electrophoresis under reducing and nonreducing conditions revealed the occurrence of two different covalently linked oligomeric complexes between these proteins, i.e., heterodimers of GP2b and GP4 and heterotrimers of GP2b, GP3, and GP4. Shortly after their release from infected cells, virions contained mainly cystine-linked GP2b/GP4 heterodimers, which were subsequently converted into disulfide-bonded GP2b/GP3/GP4 trimers through the covalent recruitment of GP3. This process occurred faster at a higher pH but was arrested at 4°C. Furthermore, the conversion was almost instantaneous in the presence of the thiol oxidant diamide. In contrast, the sulfhydryl-modifying agent N-ethylmaleimide inhibited the formation of disulfide-bonded GP2b/GP3/GP4 trimers. Using sucrose density gradients, we could not demonstrate a noncovalent association of GP3 with the cystine-linked GP2b/GP4 dimer in freshly released virions, nor did we observe higher-order structures of the GP2b/GP4 or GP2b/GP3/GP4 complexes. Nevertheless, the instantaneous diamide-induced formation of disulfide-bonded GP2b/GP3/GP4 heterotrimers at 4°C suggests that the three minor glycoproteins of EAV are assembled as trimeric complexes. The existence of a noncovalent interaction between the cystine-linked GP2b/GP4 dimer and GP3 was also inferred from coexpression experiments showing that the presence of GP3 increased the electrophoretic mobility of the disulfide-bonded GP2b/GP4 dimers. Our study reveals that the minor envelope proteins of arteriviruses enter into both covalent and noncovalent interactions, the function of which has yet to be established.
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Cerritelli, Mario E., James F. Conway, Naiqian Cheng, and Alasdair C. Steven. "Prohead Perestroika: Bacteriophage T7 Capsid Before and After Maturation." Microscopy and Microanalysis 3, S2 (August 1997): 93–94. http://dx.doi.org/10.1017/s1431927600007352.
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Bacteriophage T7 provides a well defined system to study the assembly of a moderately complex virus. The mature virion consists of a 60 nm icosahedral capsid containing the linear 40 kb genome and an internal protein “core”, and a short tail. The capsid originates as a DNA-free prohead, whose assembly requires both the major capsid protein (gpl0, ∼ 420 copies) and the scaffolding protein (gp9, ∼180 copies). Native proheads incorporate an oligomeric ring of the connector/portal protein at one vertex and the inner core, but prohead-like particles assemble when gp10 and gp9 alone are expressed. In maturation -as with other dsDNA phages - the T7 prohead undergoes a major conformational change, in which the shell structure alters dramatically, and the scaffolding protein is expelled. We have used cryo-EM in combination with other techniques to study this transition and to investigate the interactions between scaffold and surface shell in the T7 prohead.Highly purified preparations of proheads were generated from the co-expression of head and scaffolding proteins from the cloned gene in E. coli. Tail-less mature capsids as well as DNA-filled heads were prepared from infection of a T7 strain that completely lacks the tail genes.
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Rehan, Rabia, Mahrukh Kamran, Ayesha Abdul Haq, Sahar Mubeen, Maria Khan, and Hira. "Ginseng Holds Promise Against Obesity." ANNALS OF ABBASI SHAHEED HOSPITAL AND KARACHI MEDICAL & DENTAL COLLEGE 26, no. 2 (September 14, 2021): 65–70. http://dx.doi.org/10.58397/ashkmdc.v26i2.463.
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Objectives: To determine the effects of ginseng on obese albino rats.Methods: This study was intended to correlate the role of ginseng in reducing body weight, liver weight and size as well as fatty accumulation in hepatocytes of male albino rats weighing between 110g to 140g. This experiment was designed to study the morbid anatomy of an animal model. It was conducted in Animal house of Dow University of Health Sciences. Fifty male Albino Wistar rats were divided into 5 groups. Group1 (Gp 1) was on normal balanced diet (control), Group 2 (Gp 2) was on high fat diet (HFD), Group 3 (Gp 3) on HFD plus ginsenoside 100 mg/kg body wt., Group 4 (Gp 4) on HFD plus ginsenoside 200mg/kg body wt., and Group 5 (Gp 5) on HFD plus 400mg/kg body.Results: Weight was increased to 146 g (Gp1) and 236g (Gp 2). Group 3 reduced weight from 236g to 211g. Group 4 to 192g and Group 5 to 171g. Liver weight is also increased by HFD from 4.7g (Gp1) to 9.3 g (Gp 2). Liver weight decreased from 9.3g to 7.2g (Gp 3), 7.6 g (Gp 4) and 5.3 g (Gp 5). Due to deposition of fat in liver, cells enlarged and number of hepatocytes decreased per unit area of reticule. Number of hepatocytes in group 1 was 78.5, in group 2 WAS 38.3, in group 3 was 47.4, in group 4 was 53.8 andin group 5 was 67.7. Random blood sugar (RBS) was altered to 74.3 mg/dl in Gp1, 148 mg/dl in Gp2, 91 mg/dl in Gp3, 92 mg/dl in Gp4 and 69 mg/dl in Gp5.Conclusion: The results of this study revealed that HFD is a major cause of obesity and it should be prevented by introducing ginseng as an anti-obesity in our life. This study concluded that ginseng root extract proves to be more potent as anti-obesity, anti-hyperlipidemic, anti-hyperglycemic and anti-oxidant.
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Wieringa, Roeland, Antoine A. F. de Vries, Sabine M. Post, and Peter J. M. Rottier. "Intra- and Intermolecular Disulfide Bonds of theGP2b Glycoprotein of Equine Arteritis Virus: Relevance forVirus Assembly andInfectivity." Journal of Virology 77, no. 24 (December 15, 2003): 12996–3004. http://dx.doi.org/10.1128/jvi.77.24.12996-13004.2003.
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ABSTRACT Equine arteritis virus (EAV) is an enveloped, positive-strand RNA virus belonging to the family Arteriviridae of the order Nidovirales. EAV virions contain six different envelope proteins. The glycoprotein GP5 (previously named GL) and the unglycosylated membrane protein M are the major envelope proteins, while the glycoproteins GP2b (previously named GS), GP3, and GP4 are minor structural proteins. The unglycosylated small hydrophobic envelope protein E is present in virus particles in intermediate molar amounts compared to the other transmembrane proteins. The GP5 and M proteins are both essential for particle assembly. They occur as covalently linked heterodimers that constitute the basic protein matrix of the envelope. The GP2b, GP3, and GP4 proteins occur as a heterotrimeric complex in which disulfide bonds play an important role. The function of this complex has not been established yet, but the available data suggest it to be involved in the viral entry process. Here we investigated the role of the four cysteine residues of the mature GP2b protein in the assembly of the GP2b/GP3/GP4 complex. Open reading frames encoding cysteine-to-serine mutants of the GP2b protein were expressed independently or from a full-length infectious EAV cDNA clone. The results of these experiments support a model in which the cysteine residue at position 102 of GP2b forms an intermolecular cystine bridge with one of the cysteines of the GP4 protein, while the cysteine residues at positions 48 and 137 of GP2b are linked by an intrachain disulfide bond. In this model, another cysteine residue in the GP4 protein is responsible for the covalent association of GP3 with the disulfide-linked GP2b/GP4 heterodimer. In addition, our data highlight the importance of the correct association of the minor EAV envelope glycoproteins for their efficient incorporation into viral particles and for virus infectivity.
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McPhee, M. J., S. Harden, D. L. Robinson, R. W. Dicker, and V. H. Oddy. "Effects of backgrounding and finishing growth rates on P8 fat and intramuscular fat in Bos taurus steers." Animal Production Science 52, no. 5 (2012): 354. http://dx.doi.org/10.1071/an11184.
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The present study investigates the effects of backgrounding and finishing growth rates of Bos taurus steers grown out on three post-weaning growth pathways, across 4 years (1995–1998) on fat deposition. Scanned P8 fat at backgrounding exit (SP8exit), scanned P8 fat before slaughter (SP8sla) and carcass intramuscular fat (IMF, %) were evaluated. Four equations to predict SP8sla and four equations to predict carcass IMF across years were developed. Three post-weaning growth pathways (GPs) generated by different nutritional treatments across years were used in the study. Growth pathways were created by backgrounding steers on pasture with no supplementation (GP1), pasture plus supplementary feeding formulated pellets (GP2) or pasture with access to a forage crop (GP3). The steers were then finished in a feedlot or on pasture to slaughter weights of 520 kg (Korean market specification) or 600 kg (Japanese market specification). Cubic smoothing splines were fitted to the liveweight of all animals from the start of backgrounding until slaughter. Backgrounding growth rates were interpolated off the GP curves and the mean average daily gain (ADG, kg/day) at Day 100 (D100, Days 100–91 before relocation to the finishing phase), and Day 150 (D150, Days 150–141 before relocation to the finishing phase) were calculated. Significant (P < 0.01) differences between SP8exit and backgrounding ADG (BADG) were detected in all years except GP2 v. GP3 in 1997. GP1 steers had the lowest BADG across all years, ranging from 0.39 to 0.88 kg/day. At backgrounding exit, the mean SP8exit for the different GPs was ranked, with GP1 < GP3 for all years, with significant (P < 0.01) differences among GP1, GP2 and GP3 in all years except 1996. At the end of finishing, the mean carcass IMF over all years and finish (pasture and feedlot) (P < 0.05) was 4.71% for GP1 v. 5.09% for GP3 for Korean market steers and the mean SP8sla (P < 0.01) was 14.59 mm for GP1 v. 16.04 mm for GP3 and the mean carcass IMF (P < 0.01) was 5.54% for GP1 v. 6.27% for GP3 for Japanese market steers. The residual correlation of the multivariate response variables, namely, D100, D150 and BADG v. SP8exit (n = 1054), SP8sla (n = 1049) and carcass IMF (n = 1002), indicated that the highest correlations, all significant (P < 0.01), were 0.33, 0.20 and 0.12 for BADG v. SP8exit, SP8sla and carcass IMF, respectively. The residual variances (mm2) of the four SP8sla equations were 6.14, 7.44, 8.72 and 10.73 for 1995, 1996, 1997 and 1998, respectively, and the residual variances (%2) for the four carcass IMF equations were 2.51, 2.26, 1.75 and 0.84, for 1995, 1996, 1997 and 1998, respectively.
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Fan, Baochao, Hai Wang, Juan Bai, Lili Zhang, and Ping Jiang. "A Novel Isolate with Deletion inGP3Gene of Porcine Reproductive and Respiratory Syndrome Virus from Mid-Eastern China." BioMed Research International 2014 (2014): 1–11. http://dx.doi.org/10.1155/2014/306130.
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PRRSV strain SH1211 was isolated from the lung tissue of a piglet on a large-scale pig farm with approximately 30% morbidity and 50% mortality in mid-eastern China in 2012. The full-length genome of SH1211 was 15 313 nt in size, excluding the polyadenylated sequences, and shared 94.9% nucleotide sequence identity with the HP-PRRSV strain, JXA1. The GP2 and GP5 proteins of SH1211 shared only 91.5% and 85.1% amino acid sequence identities with those of the JXA1, respectively. A deletion at amino acid positions 68 and 69 was identified in the GP3 protein of SH1211, compared with the GP3 of Type-2 PRRSV isolates. A phylogenetic tree based on the nucleotide sequence of the complete genome showed that SH1211 is the most closely related to other HP-PRRSV strains isolated in China. However, phylogenetic analysis based on the GP2 and GP5 proteins showed that SH1211 is the most closely related to the QYYZ strain. A recombination analysis indicated that SH1211 might have been generated through recombination events between the JXA1 and QYYZ in which the GP2 and GP5 coding sequences were exchanged. Thus, SH1211 is a novel PRRSV strain with significant variation. Our analysis of SH1211 provides insight into the role of genetic variation in the antigenicity of PRRSVs in China.
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Sharma, Vineeta, and Renu Saxena. "Genetic Modulators: Prevalence and Their Effect on the Phenotype of HbE/β Thalassemia Patients." Blood 112, no. 11 (November 16, 2008): 1862. http://dx.doi.org/10.1182/blood.v112.11.1862.1862.
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Abstract HbE, a β-globin chain variant occurs due to GAG to AAG i.e. Glu-Lys substitution at codon 26.In heterozygous and homozygous state it usually remain asymptomatic but the compound heterozygous state with β thalassemia (HbE/β thalassemia) results in a variable and often severe anemia, with the phenotype ranging from transfusion dependence to a complete lack of symptoms. The basis of the interaction and the cause of the variability remains unexplained. Some of the possible explanations for the observed variable clinical severity are coinheritance of mild β mutations, Effect of alpha gene number and increased production of HbF. Aims of the study are to determine the frequency of primary (β mutations) and secondary modulators (α deletion and α triplication, XmnI polymorphism, HPFH-3 and Asian-Indian Inv/DelGγ(Aγδbeta;)0 deletion) in HbE/β thalassemia patients and to study their effect on the phenotype of the Patients. Subjects were 180 HbE/β thalassemia patients diagnosed by HPLC and confirmed by PCR-RFLP. Patients were divided into three subgroups according to a scoring system based on seven clinical criteria as mild (score 0–3.5)[Gp1], moderate (score 4–7)[Gp2] and severe (score 7.5–10) [Gp3]. β-mutations were studied by ARMS PCR, XmnI polymorphism was studied by PCR-RFLP while α deletions, HPFH-3 and Asian-Indian Inv/DelGγ (Aγ δβ)0 deletions were studied by GAPPCR and alpha triplication by a modified single tube PCR. The frequency of β mutation was almost similar in all the groups (IVS 1–5 G-C (58.8%) was the predominant one). α deletion was found in 19 (10.5%) out of which 13 (12 αα/−α3.7 & 1 αα /--SA) were from Gp1 and 6 (αα/-α3.7) were from Gp2. α triplication was found in 11(6.1%) out of these 11 patients 8 were (αα/αααanti3.7) from Gp3 and 3 were (αα/αααanti3.7) from Gp2. XmnI was found in 112(62.2%), out of which 98 were heterozygous (+/−) and 14 were homozygous (+/+).XmnI +/+ was present in 10 Gp1 & 4 Gp2 patients, while XmnI +/− was present in 36 Gp1 ,43 Gp2 & 33 Gp3 patients.HPFH-3 deletion was present in 8 (4.4%) out of which 5 were from Gp1 and 3 were from Gp2. Asian-Indian Inv/DelGγ (Aγδβ)0 deletion was present in 2(1.1%) patients one from Gp2 and another from GP3. Primary modulator (β mutation) is not the actual factor responsible for the disease heterogeneity. Patients with coexisting α deletion required lesser transfusions and had less severe phenotype while patients with α triplication were on frequent transfusions and had severe phenotype. XmnI polymorphism in homozygous state was observed to alleviate the severity while in heterozygous state it had no effect on the disease severity except the delay of the onset of disease up to some extent. Patients having HPFH3 deletions and Asian-Indian Inv/DelG γ (Aγδβ)0 deletion showed mild to moderate disease severity. Thus it can be concluded that secondary modulators play an important role in the disease heterogeneity of HbE/βthalassemia.
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Ismail, Ahmed M., Christoph Bourauel, Ahmed ElBanna, and Tarek Salah Eldin. "Micro versus Macro Shear Bond Strength Testing of Dentin-Composite Interface Using Chisel and Wireloop Loading Techniques." Dentistry Journal 9, no. 12 (November 30, 2021): 140. http://dx.doi.org/10.3390/dj9120140.
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Shear bond strength (SBS) testing is a commonly used method for evaluating different dental adhesive systems. Failure mode analysis provides valuable information for better interpretation of bond strength results. The aim of this study was to evaluate the influence of specimen dimension and loading technique on shear bond strength and failure mode results. Eighty macro and micro flowable composite cylindrical specimens of 1.8 and 0.8 mm diameter, respectively, and 1.5 mm length were bonded to dentinal substrate. Four study groups were created (n = 20): Macroshear wireloop, Gp1; Microshear wireloop, Gp2; Macroshear chisel, Gp3; and Microshear chisel, Gp4. They were tested for SBS using chisel and wireloop loading devices followed by failure mode analysis using digital microscopy and SEM. Two- and one-way ANOVA were used to compare stress at failure values of different groups while the Kruskal–Wallis test was used to compare between failure modes of the tested groups. Gp4 recorded the highest mean stress at failure 54.1 ± 14.1 MPa, and the highest percentage of adhesive failure in relation to the other groups. Specimen dimension and loading technique are important parameters influencing the results of shear bond strength. Micro-sized specimens and chisel loading are recommended for shear testing.
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El-Boshy, Mohamed El-Sayed, Osama Mohamed Abdalla, Angy Risha, and Fatma Moustafa. "Effect of Withania somnifera Extracts on Some Selective Biochemical, Hematological, and Immunological Parameters in Guinea Pigs Experimental Infected with E. coli." ISRN Veterinary Science 2013 (March 31, 2013): 1–6. http://dx.doi.org/10.1155/2013/153427.
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Fifty 1-2-month-old Guinea pigs were divided into 5 equal groups, 10 each. Control (Gp1) did receive neither viable bacteria nor treatment. Each animal from the other groups (Gp2–5) was challenged with (1-2 × 108) viable E. coli in 200 μL normal saline (0.9%) through IP route. GP2 infected group was treated with 200 μL saline IP and kept as positive control group. Gp3-4 are infected and treated with Withania somnifera (ethanol root extract) with doses 50 and 100 mg/kg. BW, respectively. Gp5 infected treated group was treated with cefoperazone antibiotic at dose 35 mg/Kg BW. The treatment by drug or the extracted medicinal plant was started 72 h after infection for 7 successive days. Serum and whole blood sample were collected from all groups 14 days after treatment to evaluate some hematological and biochemical changes as well as immunomodulatory cytokine tumor necrosis factor-alpha (TNF-α). Oral treatment of the plant extract caused significant benefit results in infected Guinea pig appeared in the correction of some hematological and biochemical parameters also try to suppressed inflammatory cytokine response represent in TNF-α. It could be concluded that W. somnifera extract has potent antibacterial activity, and this appears in the correction with hematological, biochemical, and immunological results.
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Stoian, Ana, and Raymond Rowland. "Challenges for Porcine Reproductive and Respiratory Syndrome (PRRS) Vaccine Design: Reviewing Virus Glycoprotein Interactions with CD163 and Targets of Virus Neutralization." Veterinary Sciences 6, no. 1 (January 17, 2019): 9. http://dx.doi.org/10.3390/vetsci6010009.
Full textAbstract:
One of the main participants associated with the onset and maintenance of the porcine respiratory disease complex (PRDC) syndrome is porcine reproductive and respiratory syndrome virus (PRRSV), an RNA virus that has plagued the swine industry for 30 years. The development of effective PRRS vaccines, which deviate from live virus designs, would be an important step towards the control of PRRS. Potential vaccine antigens are found in the five surface proteins of the virus, which form covalent and multiple noncovalent interactions and possess hypervariable epitopes. Consequences of this complex surface structure include antigenic variability and escape from immunity, thus presenting challenges in the development of new vaccines capable of generating broadly sterilizing immunity. One potential vaccine target is the induction of antibody that disrupts the interaction between the macrophage CD163 receptor and the GP2, GP3, and GP4 heterotrimer that protrudes from the surface of the virion. Studies to understand this interaction by mapping mutations that appear following the escape of virus from neutralizing antibody identify the ectodomain regions of GP5 and M as important immune sites. As a target for antibody, GP5 possesses a conserved epitope flanked by N-glycosylation sites and hypervariable regions, a pattern of conserved epitopes shared by other viruses. Resolving this apparent conundrum is needed to advance PRRS vaccine development.
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Das, Phani B., Phat X. Dinh, Israrul H. Ansari, Marcelo de Lima, Fernando A. Osorio, and Asit K. Pattnaik. "The Minor Envelope Glycoproteins GP2a and GP4 of Porcine Reproductive and Respiratory Syndrome Virus Interact with the Receptor CD163." Journal of Virology 84, no. 4 (November 25, 2009): 1731–40. http://dx.doi.org/10.1128/jvi.01774-09.
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ABSTRACT Porcine reproductive and respiratory syndrome virus (PRRSV) contains the major glycoprotein, GP5, as well as three other minor glycoproteins, namely, GP2a, GP3, and GP4, on the virion envelope, all of which are required for generation of infectious virions. To study their interactions with each other and with the cellular receptor for PRRSV, we have cloned each of the viral glycoproteins and CD163 receptor in expression vectors and examined their expression and interaction with each other in transfected cells by coimmunoprecipitation (co-IP) assay using monospecific antibodies. Our results show that a strong interaction exists between the GP4 and GP5 proteins, although weak interactions among the other minor envelope glycoproteins and GP5 have been detected. Both GP2a and GP4 proteins were found to interact with all the other GPs, resulting in the formation of multiprotein complex. Our results further show that the GP2a and GP4 proteins also specifically interact with the CD163 molecule. The carboxy-terminal 223 residues of the CD163 molecule are not required for interactions with either the GP2a or the GP4 protein, although these residues are required for conferring susceptibility to PRRSV infection in BHK-21 cells. Overall, we conclude that the GP4 protein is critical for mediating interglycoprotein interactions and, along with GP2a, serves as the viral attachment protein that is responsible for mediating interactions with CD163 for virus entry into susceptible host cell.
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Savoia, Anna, Shinji Kunishima, Patrizia Noris, Nuria Pujol-Moix, Dermot Kenny, Nurit Rosenberg, Margaret L. Rand, et al. "International Consortium for the Study of Clinical and Molecular Aspects of Bernard-Soulier Syndrome." Blood 118, no. 21 (November 18, 2011): 707. http://dx.doi.org/10.1182/blood.v118.21.707.707.
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Abstract Abstract 707FN2 Bernard-Soulier syndrome (BSS) is an extremely rare inherited bleeding disorder characterized by a defect of the GPIb/IX/V complex, which is essential for hemostasis, as the GPIbα subunit binds to subendothelial von Willebrand factor. Since the identification of the first mutation in 1990, almost one hundred cases carrying mutations in the GP1BA, GP1BB, and GP9 genes have been described. Most of the mutations prevent the coordinated association of the complex or binding to the von Willebrand factor. BSS is usually transmitted as a recessive trait with giant platelets and severe bleeding tendency. However, there are families with a dominant mild form, in which the affected individuals have only moderate macrothrombocytopenia and bleeding tendency. A correct definition of the clinical and laboratory features, together with accurate genotype/ phenotype correlation studies, remains essential for understanding the molecular basis of the disease and managing patients appropriately. Moreover, it is important to understand the variability of clinical manifestations. Since BSS is rare with an estimated prevalence of 1:1,000,000, an International Consortium has recently been established to collect a large series of cases and families worldwide. At present, the Consortium has been compiling data from 165 unrelated families, of which 50% have not been previously described. In this cohort, the molecular genetic testing reveals more than 30 novel mutations, confirming the wide spectrum of alterations responsible for the disease. Data from 65 unrelated families (69 patients) mainly from France, Italy and Japan show that 23 have mutations in GP9 and 29 in GP1BB. In the remaining 13 families, the defective gene is GP1BA. In agreement with the view that BSS is an extremely rare disease, 53 probands carried homozygous mutations, 10 are compound heterozygous, and 2 hemizygous because of a 22q11 deletion of the DiGeorge syndrome. The mean age of patients at diagnosis was 18 years (range 0–75 years) of which 27 were males and 38 females. Misdiagnosis of autoimmune thrombocytopenia was frequent and 26 patients were previously treated with steroids, intravenous immunoglobulins and/or splenectomy. Except two Japanese cases without any bleeding manifestations, patients presented with a variable bleeding diathesis measured by the World health Organization bleeding scale: grades 1, 2, 3 and 4 in 9, 18, 19 and 10 patients, respectively. The mean platelet count was 64×109/L (range 24–130) as determined by microscopy. In contrast, using a cell counter, thrombocytopenia was more severe (45×109/L; range 5–125). The mean platelet mean diameter was larger than in controls and varied from 2.9 to 7.5 mm. Ristocetin-induced platelet agglutination was absent or lower than 22% of normal response in all patients. Flow cytometry revealed a defective expression of the GPIb/IX/V expression in all patients. Correlating between expression data and gene affected, we found that the expression of GP1ba was often undetectable in patients with GP1BA mutations whereas it was higher, 8% and 17%, in patients with mutations of GP1BB and GP9, respectively. Instead, the GPIX mean level was 14%, 8% and 25% in patients with GP9, GP1BB and GP1BA mutations. The expression of GPV was higher than that of the other subunits, being more than 30% regardless of which gene was mutated. This is the largest cohort of BSS patients characterized to date. These patients together with the other 100 cases not yet included in the BSS database will enable correlations of the molecular genetic defects, receptor expression and clinical manifestations observed in BSS patients. Disclosures: Zieger: CSL Behring Hattersheim: Research Funding.
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Dave, Janam J., Adilene Sandoval, Jon Olson, and Jill Adler-Moore. "1577. Particle Characterization of Nebulized Liposomal Amphotericin B and Its Use in the Treatment of Murine Pulmonary Aspergillosis." Open Forum Infectious Diseases 6, Supplement_2 (October 2019): S576. http://dx.doi.org/10.1093/ofid/ofz360.1441.
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Abstract Background Immunocompromised patients are very susceptible to pulmonary aspergillosis causing 50% mortality with present treatments, indicating a need for improved therapy. To address this, we standardized a nebulization method for effectively delivering liposomal amphotericin B (AmBisome®, AmBi) into lungs of Aspergillus fumigatus-infected mice. Methods AmBi particle characterization was done with a Cascade particle impactor and a Schuco S5000 nebulizer containing 1.33 mg/mL AmBi. For in vivo studies, AmBi was nebulized (neb) into a 12 compartment chamber (one mouse/compartment), following immunosuppression with 28 mg/kg triamcinolone IP (d-3, -1, +1). Mice were challenged d0 with 9 x 106A. fumigatus (ATCC#13073) and 4 hours post-challenge, divided into 5 groups (n = 12/gp): 5 days of 20 min/day neb AmBi (Gp1), 5 days of 10 min/day neb AmBi (Gp2), 20 min/day neb AmBi days 0, 1, 3, 5, 7 (Gp 3), 5 days of intravenous(IV) AmBi 7.5 mg/kg/day (Gp4) and IV PBS (Gp5). Seven mice/gp were monitored for survival to d21 and lungs, livers, kidneys, spleens (5 mice/gp) analyzed for mean amphotericin B µg/g and CFU/g. Results 87% of neb AmBi particles were between 0.43 mm to 3.3 mm allowing for drug penetration into 1°, 2° and terminal bronchi, bronchioles, and alveoli. This resulted in very good protection, with 20 min daily neb treatments (Gp1) giving 100% survival and 10 min daily neb treatments producing 71% survival (Gp2). There were no survivors in the PBS gp (P < 0.02 vs. Gp1 and Gp2). Every other day neb AmBi or daily IV AmBi was less effective (43% survival). In addition, neb AmBi for 20 min (Gp1) yielded significantly lower fungal burden in lungs vs. all other AmBi treatments (P < 0.02). While drug was detected in lungs of mice given 20 min of neb AmBi (2.6 µg/g), there was no drug detected in livers, kidneys or spleens of any mice given neb AmBi. In comparison, with IV AmBi, drug was detected in the lungs (7 µg/g), livers (204 µg/g), kidneys (38 µg/g), and spleens (114 µg/g). Conclusion Daily AmBi nebulization was an effective and potentially less nephrotoxic treatment for murine pulmonary aspergillosis since it achieved significantly lower tissue fungal burden and much better survival vs. daily IV AmBi, without delivering drug to the kidneys. Disclosures All authors: No reported disclosures.
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Salem, Siham, Sabry Naggar, Ramadan Kandyel, and Marwa Sharkawy. "Histological and immunohistochemical studies on the effect of zinc oxide nanoparticles on the cerebellum of newborn and adult rats and the possible protective role of EDTA." EGYPTIAN JOURNAL OF EXPERIMENTAL BIOLOGY (Zoology) 19, no. 1 (2023): 15. http://dx.doi.org/10.5455/egysebz.20221222014955.
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Zinc oxide nanoparticles (ZnO NPs) are involved in different applications such as nutrition, while, it causes damage to different organs of the body including the brain. This study aimed to compare between the effect of ZnO NPs on newborn and adult rats´ cerebellum and to evaluate the protective role of Ethylenediaminetetraacetic acid (EDTA) against ZnO NPs toxicity. Thirty-two newborn (NBM)and thirty-two adult male (ADM) Sprague Dawley rats are used. Four groups (n = 8) of each are divided as following; Gp1: is served as control, Gp2: injected intraperitoneal (IP) with ZnO NPs (3 g/ kg). Gp3: injected IP with EDTA (500 mg/ kg) for 7 days. Gp4: injected with ZnO NPs and treated with EDTA as in Gp3. Brain tissue is prepared for biochemical and histopathological investigations. Newborn and adult rats injected with ZnO NPs showed degeneration of purkinje and granular cells in cerebellum tissue confirmed with high expression of caspase-3, while, treatment with EDTA post ZnO NPs showed lightly improvement. The body weights and relative brain weights is decrease in all the rats treated with ZnO NPs. In addition, our current study proved that the effect of ZnO NPs on ADM rats is more than on the NBM ones. Collectively, ZnO NPs injection caused damage of the cerebellum in newborn and adult rats, however, the treatment with EDTA stopped somewhat this damage.
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Matczuk, Anna Karolina, Grzegorz Chodaczek, and Maciej Ugorski. "Production of Recombinant EAV with Tagged Structural Protein Gp3 to Study Artervirus Minor Protein Localization in Infected Cells." Viruses 11, no. 8 (August 9, 2019): 735. http://dx.doi.org/10.3390/v11080735.
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Equine arteritis virus (EAV) is a prototype member of the Arterivirus family, comprising important pathogens of domestic animals. Minor glycoproteins of Arteriviruses are responsible for virus entry and cellular tropism. The experimental methods for studying minor Arterivirus proteins are limited because of the lack of antibodies and nested open reading frames (ORFs). In this study, we generated recombinant EAV with separated ORFs 3 and 4, and Gp3 carrying HA-tag (Gp3-HA). The recombinant viruses were stable on passaging and replicated in titers similar to the wild-type EAV. Gp3-HA was incorporated into the virion particles as monomers and as a Gp2/Gp3-HA/Gp4 trimer. Gp3-HA localized in ER and, to a lesser extent, in the Golgi, it also co-localized with the E protein but not with the N protein. The co-localization of Gp3-HA and the E protein with ERGIC was reduced. Moreover, EAV with Gp3-HA could become a valuable research tool for identifying host cell factors during infection and the role of Gp3 in virus attachment and entry.
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Adair, W. S., S. A. Steinmetz, D. M. Mattson, U. W. Goodenough, and J. E. Heuser. "Nucleated assembly of Chlamydomonas and Volvox cell walls." Journal of Cell Biology 105, no. 5 (November 1, 1987): 2373–82. http://dx.doi.org/10.1083/jcb.105.5.2373.
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The Chlamydomonas reinhardtii cell wall is made up of hydroxyproline-rich glycoproteins, arranged in five distinct layers. The W6 (crystalline) layer contains three major glycoproteins (GP1, GP2, GP3), selectively extractable with chaotropic agents, that self-assemble into crystals in vitro. A system to study W6 assembly in a quantitative fashion was developed that employs perchlorate-extracted Chlamydomonas cells as nucleating agents. Wall reconstitution by biotinylated W6 monomers was monitored by FITC-streptavidin fluorescence and quick-freeze/deep-etch electron microscopy. Optimal reconstitution was obtained at monomer concentrations (0.2-0.3 mg/ml) well below those required for nonnucleated assembly. Assembly occurred from multiple nucleation sites, and faithfully reflected the structure of the intact W6 layer. Specificity of nucleated assembly was demonstrated using two cell-wall mutants (cw-2 and cw-15); neither served as a substrate for assembly of wild-type monomers. In addition, W6 sublayers were assembled from purified components: GP2 and GP3 coassembled to form the inner (W6A) sublayer; this then served as a substrate for self-assembly of GP1 into the outer (W6B) sublayer. Finally, evolutionary relationships between C. reinhardtii and two additional members of the Volvocales (Chlamydomonas eugametos and Volvox carteri) were explored by performing interspecific reconstitutions. Hybrid walls were obtained between C. reinhardtii and Volvox but not with C. eugametos, confirming taxonomic assignments based on structural criteria.
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Thi Nga, Nguyen, Ha Thi Thu, Nguyen Thi Hoa, Vu Thi Hien, Nguyen Thu Trang, Nguyen Thanh Ba, Tran Van Khanh, et al. "Assessment of the genetic changes of the attenuated Hanvet1.vn strain compared with original virulent 02HY strain of the porcine reproductive and respiratory syndrome virus." Vietnam Journal of Biotechnology 20, no. 2 (June 30, 2022): 245–52. http://dx.doi.org/10.15625/1811-4989/16677.
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The attenuated porcine reproductive and respiratory syndrome virus (PRRSV) strain Hanvet1.vn was developed by Hanvet Pharmaceutical Co., Ltd. by inoculating the virulent strain 02HYon Marc-145 cells for 80 generations and used to produce PRRS vaccine. In this study, we published the results of sequencing, analyzing and comparing the genome of the attenuated PRRSV strain Hanvet1.vn compared with the original pathogenic strain 02HY. The genomes of strains Hanvet1.vn and 02HY have 8 reading frames, coding for 8 non-structural and structural proteins: NSP1a, NSP1b, GP2, GP3, GP4, GP5, MP, NP. After sequencing and translating into proteins, the gene sequence of each open reading frame (ORF) of strain Hanvet1.vn was compared with the sequence of pathogenic strain 02HY to find nucleotide and amino acid changes. The results showed that the Hanvet1.vn pathogenic strain genome (Genbank Accession KU842720) when compared with the pathogenic strain 02HY genome (Submission2490633) had89 nucleotide mutations that changed 51 amino acids in 7 ORFs and 7 proteins, respectively. Particularly, ORF6 encoding for the M protein is completely unchanged. The size of each reading frame is also exactly the same. It showed that there were no insertion and deletion (Indel) mutations in the ORFs of the attenuated strain after 80 generations of inoculation. There was a change in the genome that made the strain Hanvet1.vn become attenuated, but the gene encoding for the GP5 protein that induces the production of neutralizing antibodies only changed two nucleotides at position 471 (A->G), causing the TCA codon to turn into a TCG codon. This is a silent mutation and both codons code for the amino acid Serine (S). The second mutation at position 587 (A->T) causes Glutamine (Q) to transform into Leucine (L). However, this modification does not belong to the GP5 antigenic epitopes. In clonclusion, after 80 passages, despite changes occurred in genes of Hanvet1.vn strain for becoming an attenuated strain, the GP5 protein of the attenuated strain did not change its antigenic amino acids.
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Bartoszko, Justyna, Tony Panzerella, Anthea Lau, Naheed Alam, Aaron D. Schimmer, Matthew D. Seftel, Andre C. Schuh, Mohamed Shanavas, Karen W. L. Yee, and Vikas Gupta. "Comparison of the Impact of Two Different Definitions of Red-Cell Transfusion Dependence on the Natural History of Myeloproliferative Neoplasm (MPN)-Associated Myelofibrosis (MF)." Blood 124, no. 21 (December 6, 2014): 3180. http://dx.doi.org/10.1182/blood.v124.21.3180.3180.
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Abstract The current definitions of red-cell transfusion dependence (TD) for MPN-associated MF are based on expert opinion. The optimal definition of TD, and its impact on the natural history of MF is not well studied. We evaluated the impact of two definitions of red-cell TD on the natural history of 306 patients (pts.) with WHO/IWG-MRT-defined primary MF (PMF, n=202, 66%), post-polycythemia vera MF (PPV-MF, n=45, 15%) or post-essential thrombocythemia MF (PET-MF, n=59, 19%). Red-cell TD was defined according to IWG-MRT 2006 criteria (Tefferi et al, Blood, 2006), and on the results of the expert consensus RAND-Delphi panel (Gale et al, Leukemia Research, 2011). The IWG-MRT criteria require patients to have received ≥2 Units (U) packed red blood cells (PRBC) in the preceding 28 days for hemoglobin (Hb) <85 g/l. On the other hand, the more stringent Gale definition requires patients to receive ≥2 U of PRBC per month over 3 months without any specification for the Hb level. Patients were evaluated for disease risk stratification according to dynamic international prognostic scoring system (DIPSS), co-morbidities according to ACE-27, and red-cell TD at their first presentation to Princess Margaret Cancer Center or diagnosis. The study cohort was divided in 4 groups (gp.): Non-transfusion dependent (NTD) pts. with Hb ≥100 g/dl (gp1, n=156, 51%); patients with Hb <100 g/dl, but no transfusion in preceding 12 weeks (gp2, n=65, 21%); pts. qualifying IWG-MRT 2006 criteria of TD, but not qualifying Gale definition (gp3, n=41, 13%); and pts. qualifying the Gale definition of TD (gp4, n=44, 14%). There was no significant difference among the 4 groups with relation to age, disease distribution (primary vs. secondary), cytogenetics, blasts%, and ACE-27 co-morbidity scores. There was significant difference in the gp1, gp2, gp 3, and gp4 in relation to constitutional symptoms (35% vs. 45% vs. 46% vs.64%, p=0.006), median WBC count (x109/L) (11.5 vs. 8.3 vs. 6.8 vs. 6.5, p<0.0001), platelets (x109/l) (317 vs. 154 vs. 124 vs. 127, p<0.0001), and Int-2/high DIPSS (24% vs. 82% vs. 90% vs. 82%, p<0.0001). In a univariate analysis, probability of survival among the 4 groups at 3-years was 80%, 55%, 68%, 30%, respectively (Fig 1, p=0.0016). No difference in leukemic transformation was observed among the 4 groups (p=0.68). In a multivariate analysis, after adjusting for DIPSS and co-morbidity scores, hazard ratios among the gp2, gp3, and gp 4 were 1.58 (95% CI 0.78-3.19, p=0.21), 2.17 (95% CI 0.88-5.34, p=0.09), 2.82 (95% CI 1.29-6.16, p=0.009) in comparison to gp 1 (ref. group). There was no significant difference between NTD patients with Hb <100 vs. TD patients according to IWG-MRT 2006 criteria, but not meeting Gale Criteria (p=0.47). DIPSS was the only other independent prognostic factor for survival. Our study shows the independent prognostic impact of TD on survival defined according to Gale criteria, but not according to previous IWG-MRT 2006 definition, which does not meet Gale criteria. No impact of anemia or TD was observed on leukemic transformation. Fig 1: Survival in 4 groups according to Hb levels and transfusion needs Fig 1:. Survival in 4 groups according to Hb levels and transfusion needs Disclosures Schuh: Celgene: Membership on an entity's Board of Directors or advisory committees.
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de Siqueira, Lucia Helena, Miriam P. Beltrame, Fernanda P. G. Cunha, Marina P. Colella, Joyce M. Annichino-Bizzacchi, Erich V. de Paula, and Margareth C. Ozelo. "Six Novel Mutations Identified in the Glycoproteins Ib Alpha, Ib Beta and IX Genes Among Twenty-Two Unrelated Patients with Bernard-Soulier Syndrome in Brazil." Blood 118, no. 21 (November 18, 2011): 1156. http://dx.doi.org/10.1182/blood.v118.21.1156.1156.
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Abstract Abstract 1156 Introduction: Bernard-Soulier syndrome (BSS) is a rare hereditary bleeding disorder, caused by mutations within the glycoprotein (GP) Ib alpha, GP Ib beta and GP IX genes that encode three of four subunits of the platelet GP Ib-V-IX adhesion receptor. In the present study we evaluated the mutations involved in the diagnosis of BSS from twenty-two unrelated patients. Patients and Methods: All patients were followed in two large bleeding disorder reference centers in Brazil. The diagnosis of BSS was established based on the presence of mucocutaneous bleeding and macrotrombocytopenia, and was confirmed by platelet ristocetin aggregation and flow cytometry for the platelet GP Ib alpha (CD 42b), GP Ib beta (CD 42c), and GP IX (CD 42a). Available first- and second-degree relatives were also contacted for clinical, laboratory and molecular evaluation. Genomic DNA from all index cases was used for sequence analysis of the three genes, GP1BA, GP1BB and GP9, and the results were confirmed in relatives when available. Results: Twenty-two unrelated patients with the confirmed diagnosis of BSS were enrolled in this study. Among twenty-two index cases, twenty-one had one or two mutations identified, including six novel mutations. We also identified two mutations that have been previously reported, GP1BA C209S (Simsek, et al., 1994), and GP9 A140T (Wang, et al., 2004). The six novel mutations correspond to conserved regions, and they consist of two mutations in the GP1BA (L51R, and L99P), three in GP1BB (M-25A, L72R, and L112P), and one in GP9 (P52Q). One of these novel mutations, the GP1BB L112P was observed in twelve of twenty-two unrelated cases. PCR-restriction fragment length analysis of genomic DNA from a hundred normal unrelated controls were performed to evaluate the presence of GP1BA L99P and GP1BB L112P mutations by using the AlwN I and Alu I restriction enzymes, respectively. All controls were negative for both mutations. Interestingly, when we analyzed the relatives of the index cases indentified with the mutations GP1BA L99P and GP1BB M-25A, we found evidence of mild macrotrombocytopenia in the heterozygote carriers of these mutations. The relatives heterozygous for GP1BB L112P showed normal platelet count and morphology. However, in three index cases with mild macrotrombocytopenia, GP1BB L112P in heterozygosity was the only mutation identified. Furthermore, site-directed mutagenesis was carried out to evaluate the expression of the novel mutations GP1BA L51R, and GP1BA L99P. Compared to the GP1BA wild-type, both mutations were only minimally expressed in CHO bIX cells, which stably express GP Ib beta and GP IX. Conclusions: We identified in this study eight distinct mutations among twenty-two unrelated SBS patients, including six novel mutations. The GP1BB L112P mutation was found in twelve of the twenty-two index cases, suggesting that this could be due to a founder effect. Identifying such a frequent mutation in this population of BSS patients will be helpful for genetic diagnosis of this condition in Brazil. Furthermore these mutations significantly add to the mutation database of BSS and will inevitably provide insights into the function of GP Ib-V-IX. Disclosures: No relevant conflicts of interest to declare.
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I. Ciftci, Halil, Mohamed O. Radwan, Safiye E. Ozturk, N. Gokce Ulusoy, Ece Sozer, Doha E. Ellakwa, Zeynep Ocak, et al. "Design, Synthesis and Biological Evaluation of Pentacyclic Triterpene Derivatives: Optimization of Anti-ABL Kinase Activity." Molecules 24, no. 19 (September 30, 2019): 3535. http://dx.doi.org/10.3390/molecules24193535.
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Imatinib, an Abelson (ABL) tyrosine kinase inhibitor, is a lead molecular-targeted drug against chronic myelogenous leukemia (CML). To overcome its resistance and adverse effects, new inhibitors of ABL kinase are needed. Our previous study showed that the benzyl ester of gypsogenin (1c), a pentacyclic triterpene, has anti-ABL kinase and a subsequent anti-CML activity. To optimize its activities, benzyl esters of carefully selected triterpenes (PT1–PT6), from different classes comprising oleanane, ursane and lupane, and new substituted benzyl esters of gypsogenin (GP1–GP5) were synthesized. All of the synthesized compounds were purified and charachterized by different spectroscopic methods. Cytotoxicity of the parent triterpenes and the synthesized compounds against CML cell line K562 was examined; revealing three promising compounds PT5, GP2 and GP5 (IC50 5.46, 4.78 and 3.19 μM, respectively). These compounds were shown to inhibit extracellular signal-regulated kinase (ERK) downstream signaling, and induce apoptosis in K562 cells. Among them, PT5 was identified to have in vitro activity (IC50 = 1.44 μM) against ABL1 kinase, about sixfold of 1c, which was justified by molecular docking. The in vitro activities of GP2 and GP5 are less than PT5, hence they were supposed to possess other more mechanisms of cytotoxicity. In general, our design and derivatizations resulted in enhancing the activity against ABL1 kinase and CML cells.
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White, I. N. H., D. C. Blakey, M. L. Green, M. Jarman, and H. R. Schulten. "Factors responsible for the formation of different N-alkylated porphyrins in rat liver microsomal systems exposed to norethindrone. The role of 3 α-hydroxysteroid dehydrogenase." Biochemical Journal 236, no. 2 (June 1, 1986): 379–87. http://dx.doi.org/10.1042/bj2360379.
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Incubation of rat liver microsomes with norethindrone and a NADPH-generating system leads to the formation of one N-alkylated porphyrin (green pigment, GP1). Administration of this steroid to male rats in vivo results in the formation of three more-polar green pigments (GP2, 3 and 4). A cytosolic protein (green-pigment converting protein) has been purified from rat liver that, when added to liver microsomal mixtures containing norethindrone (0.03 mM) and a NADPH-generating system, results in the formation of all four green pigments (GP1, 2, 3 and 4). Field-desorption mass spectrometry of the purified green pigments gave protonated molecules, [M + H]+, at m/z 905 for GP1, m/z 909 for GP2, m/z 925 for GP3 and 4. The Mr of the purified cytosolic protein on SDS/polyacrylamide-gel electrophoresis or gel filtration was 37000. Polyacrylamide-gel isoelectric focusing gave a pI value of 5.9. Antibodies raised in rabbits against this protein, after preincubation with rat liver cytosol, subsequently prevented the formation of the more-polar norethindrone-induced green pigments (GP2, 3 and 4). The purified protein in the presence of either NADH or NADPH catalysed the reduction of delta 4-ring-reduced norethindrone, 5 alpha-oestran-17 alpha-ethynyl-17 beta-ol-3-one and, with the appropriate cofactor, the oxidation and reduction of steroids lacking the ethynyl function, e.g. androsterone or dihydrotestosterone. Indomethacin inhibited the reduction of dihydrotestosterone by this protein with an I50 (concn. causing 50% inhibition) value of 4.9 microM. From its physical and enzymic properties it is concluded that green-pigment converting protein is the same as 3 alpha-hydroxysteroid dehydrogenase (EC 1.1.1.50).
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GOODENOUGH, URSULA W., and JOHN E. HEUSER. "Molecular organization of cell-wall crystals from Chlamydomonas reinhardtii and Volvox carteri." Journal of Cell Science 90, no. 4 (August 1, 1988): 717–34. http://dx.doi.org/10.1242/jcs.90.4.717.
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The extracellular matrices of Chlamydomonas reinhardtii and Volvox carteri contain homologous salt-extractable crystalline layers that will self-assemble in vitro. The organization of these crystals is examined using the quick-freeze deepetch technique. In C. reinhardtii, the outer layer of the crystal is an open polygonal weave (W6B); this is shown to be constructed from regular overlapping associations between the fibrous hydroxyproline-rich glycoprotein GP1. The inner layer of the crystal (W6A) is shown to be a copolymer of GP2 and GP3. The bulky globular domains of these glycoproteins form the rows of granules seen on the upper and lower surfaces of W6A in both Chlamydomonas and Volvox, and their filamentous domains interact to form a dense meshwork.
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Piuri, Mariana, Liliana Rondón, Estefanía Urdániz, and Graham F. Hatfull. "Generation of Affinity-Tagged Fluoromycobacteriophages by Mixed Assembly of Phage Capsids." Applied and Environmental Microbiology 79, no. 18 (July 12, 2013): 5608–15. http://dx.doi.org/10.1128/aem.01016-13.
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ABSTRACTAddition of affinity tags to bacteriophage particles facilitates a variety of applications, including vaccine construction and diagnosis of bacterial infections. Addition of tags to phage capsids is desirable, as modification of the tails can lead to poor adsorption and loss of infectivity. Although tags can readily be included as fusions to head decoration proteins, many phages do not have decoration proteins as virion components. The addition of a small (10-amino-acid) Strep-tag II (STAG II) to the mycobacteriophage TM4 capsid subunit, gp9, was not tolerated as a genetically homogenous recombinant phage but could be incorporated into the head by growth of wild-type phage on a host expressing the capsid-STAG fusion. Particles with capsids composed of wild-type and STAG-tagged subunit mixtures could be grown to high titers, showed good infectivities, and could be used to isolate phage-bacterium complexes. Preparation of a STAG-labeled fluoromycobacteriophage enabled capture of bacterial complexes and identification of infected bacteria by fluorescence.
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Kim, Pil Soo, Yeo-Rang Lee, Yong-Su Kwon, Jin-Woo Bae, Sung-Jae Lee, and Young-Seuk Park. "Differences of Gut Microbiota in the Freshwater Blackworm (Lumbriculus variegatus: Oligochaeta) in Two Different Habitat Conditions." International Journal of Environmental Research and Public Health 18, no. 19 (September 29, 2021): 10298. http://dx.doi.org/10.3390/ijerph181910298.
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The distribution of organisms is governed by their habitat condition. We analyzed bacterial communities in the gut of the blackworm Lumbriculus variegatus by pyrosequencing of the extracted intestinal metagenomic DNA. Blackworms were collected from two sampling sites with differences in irradiance and riparian vegetation, where site GP7 was covered by riparian vegetation and site GP8 was exposed to sunlight. We obtained the filtered 6414 reads from three samples of each site. At GP7, 271 OTUs were identified, including 32 OTUs unique to the site, whereas at GP8, 238 OTUs were identified, including 22 unique OTUs. Among them, 18 OTUs were shared between both sites. The phylum Proteobacteria was a major component contributing 67.84% and 64.05% of sequences at sites GP7 and GP8, respectively, while each remaining phylum contributed less than 10% at both sites. The two sites differed in microbial community composition and KEGG-indicated biochemical pathways. Community indices such as species richness and Shannon diversity were higher at site GP7 than at GP8. Meanwhile, the abundance of Cyanobacteria was significantly higher at site GP8, while site GP7 showed a greater proportion of genes for membrane transport and carbohydrate metabolism, reflecting differences in food resources.
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Jiang, M., ML Mei, MCM Wong, CH Chu, and ECM Lo. "Influence of Silver Diamine Fluoride Treatment on the Microtensile Bond Strength of Glass Ionomer Cement to Sound and Carious Dentin." Operative Dentistry 45, no. 5 (June 5, 2020): E271—E279. http://dx.doi.org/10.2341/19-237-l.
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Clinical Relevance This study provides valuable information about the influence of silver diamine fluoride (SDF) treatment on the microtensile bond strength of glass ionomer cement (GIC) to dentin. SUMMARY Objectives: To investigate the influence of silver diamine fluoride (SDF) treatment on the microtensile bond strength (mTBS) of glass ionomer cement (GIC) to sound and artificial carious dentin. Methods: Thirty dentin blocks prepared from 30 noncarious human molars were randomly allocated into either the sound (Gp1) or artificial carious dentin (Gp2) groups. A microbiological method was adopted to create artificial dentin caries lesions in Gp2 specimens. Each dentin block was sectioned into two halves perpendicularly, and each pair of block halves was randomly assigned to two subgroups to receive topical application of SDF (Gp1-SDF, Gp2-SDF) or water as control (Gp1-water, Gp2-water). An encapsulated GIC was bonded to the exposed dentin surfaces 14 days after the SDF/water application. After immersion for 7 days in artificial saliva, the GIC-dentin specimens were sectioned into beams for mTBS testing. Failure mode was examined after the mTBS test. Results: There was no significant difference in the mean mTBS values between the SDF and control subgroups (Gp1-SDF vs Gp1-water, 10.57±1.6 MPa vs 10.20±1.8 MPa; Gp2-SDF vs Gp2-water, 6.14±2.2 MPa vs 5.97±2.3 MPa; paired t-test, p>0.05). However, the mean mTBS value of the sound dentin group was significantly higher than that of the carious dentin group, irrespective of whether SDF was applied prior to GIC bonding (independent t-test, p<0.001). Proportionally more cohesive failures occurred in the sound dentin groups (Gp1-SDF, 48.4%; Gp1-water, 42.9%) compared with the carious dentin groups (Gp2-SDF, 15.6%; Gp2-water, 9.8%; p<0.05). Conclusions: SDF treatment had no significant influence on the mTBS of GIC to dentin. Compared with sound dentin, dentin with caries had lower mTBS to GIC.
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Norelli, Jolanta B., Dawid P. Plaza, Drew N. Stal, Anish M. Varghese, Haixiang Liang, and Daniel A. Grande. "Tenogenically differentiated adipose-derived stem cells are effective in Achilles tendon repair in vivo." Journal of Tissue Engineering 9 (January 2018): 204173141881118. http://dx.doi.org/10.1177/2041731418811183.
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The purpose of this study was to characterize rat adipose-derived stem cells, induce adipose-derived stem cell tenogenesis, and analyze adipose-derived stem cell effects on tendon repair in vivo. Adipose-derived stem cells demonstrated an immunomodulatory, pro-angiogenic, and pro-proliferatory profile in vitro. Tenogenesis was induced for 1, 7, 14, and 21 days with 24 combinations of growth differentiation factor-5, 6, and 7 and platelet-derived growth factor–BB. Adipose-derived stem cells expression of scleraxis and collagen type I increased the most after 14 days of induction with growth differentiation factor-6 and platelet-derived growth factor–BB. Achilles excision defects injected with hydrogel alone (Gp2), with undifferentiated (Gp3) adipose-derived stem cells, or tenogenically differentiated (Gp4) adipose-derived stem cells exhibited improved tissue repair compared with untreated tendons (Gp1). Addition of adipose-derived stem cells improved tissue cytoarchitecture and increased expression of collagen type I and III, scleraxis, and tenomodulin. Adipose-derived stem cells significantly improved biomechanical properties (ultimate load and elastic toughness) over time more than hydrogel alone, while tenogenically differentiated adipose-derived stem cells improved the mean histological score and collagen fiber dispersion range closest to normal tendon. In addition, tendon sections treated with GFP-adipose-derived stem cells exhibited green fluorescence and positive GFP immunostaining on microscopy confirming the in vivo survival of adipose-derived stem cells that were injected into tendon defects to support the effects of adipose-derived stem cells on tissue up to 4.5 weeks post injury.
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Dolci, E. D., and G. E. Palade. "Ontogenetic expression of the murine erythrocyte glycophorins." Journal of Cell Science 93, no. 1 (May 1, 1989): 191–97. http://dx.doi.org/10.1242/jcs.93.1.191.
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The major sialoglycoproteins or glycophorins of the murine erythrocyte membrane, gp2 (Mr 44,000) and gp3 (Mr 29,000), are expressed as identical or closely related antigens by the three erythroid cell lines that succeed each other in normal mouse development. The embryonic forms of gp2 and gp3 differ from the adult forms by their mobility on SDS-PAGE. The apparent Mr values are increased by 1000–2000 for gp2 and 500 for gp3. An increase in the microheterogeneity of both embryonic and fetal forms of gp2 is also detected. Variations in glycosylation account in part for the apparent differences in Mr. Sizing of O-glycosidically linked oligosaccharide chains reveals that fetal glycophorins contain predominantly trisaccharide units while their adult counterparts are mostly tetrasaccharides. The kinetics of gp2 and gp3 biosynthesis in fetal liver are comparable to those established for the splenic erythroblasts of adult anemic mice. Like their adult counterparts, fetal glycophorins incorporate [3H]palmitate and [3H]galactose. The results indicate that, in murine ontogeny, distinct but antigenically related sialoglycoproteins are produced at each erythropoietic stage.
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Mmolawa, Princess T., Horst Schmieger, Carly P. Tucker, and Michael W. Heuzenroeder. "Genomic Structure of the Salmonella enterica Serovar Typhimurium DT 64 Bacteriophage ST64T: Evidence for Modular Genetic Architecture." Journal of Bacteriology 185, no. 11 (June 1, 2003): 3473–75. http://dx.doi.org/10.1128/jb.185.11.3473-3475.2003.
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ABSTRACT The complete sequence of the double-stranded DNA genome of a serotype-converting temperate bacteriophage, ST64T, was determined. The 40,679-bp genomic sequence of ST64T has an overall GC content of 47.5% and was reminiscent of a number of lambdoid phages, in particular, P22. Inferred proteins of ST64T which exhibited a high degree of sequence similarity to P22 proteins (>90%) included the functional serotype conversion cassette, integrase, excisionase, Abc1, Abc2, early antitermination (gp24), NinD, NinH, NinZ, packaging (gp3 and gp2), head (with the exception of gp26, gp7, gp20, and gp16), and tail proteins. The putative immunity genes were highly related to those of Salmonella enterica serotype Typhimurium phage L, whereas the lysis genes were almost identical to those of S. enterica serovar Typhimurium PS3.
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Ploss, Martin, and Andreas Kuhn. "Membrane insertion and assembly of epitope-tagged gp9 at the tip of the M13 phage." BMC Microbiology 11, no. 1 (2011): 211. http://dx.doi.org/10.1186/1471-2180-11-211.
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Gentilini, Fabio, Maria Elena Turba, Fiorella Giancola, Roberto Chiocchetti, Chiara Bernardini, Markéta Dajbychova, Vidhya Jagannathan, Michaela Drögemüller, and Cord Drögemüller. "A large deletion in the GP9 gene in Cocker Spaniel dogs with Bernard-Soulier syndrome." PLOS ONE 14, no. 9 (September 4, 2019): e0220625. http://dx.doi.org/10.1371/journal.pone.0220625.
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Ivers, D. J. "On generalised toroidal-poloidal solutions of vector field equations." Journal of the Australian Mathematical Society. Series B. Applied Mathematics 30, no. 4 (April 1989): 436–49. http://dx.doi.org/10.1017/s0334270000006378.
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AbstractThe orthogonal coordinate systems ξi(i = 1, 2, 3) are determined, in which the gneralised toroidal and poloidal fields, defined respectively by T{T} = ∇ × {T∇ξ1} and S{S} = ∇ × T{S}, have the following three properties:GP1 Decoupling of the vector Helmholtz equation: There exist linear differential operators L1 and L2 such that Hu = 0, where H is the vector Helmholtz operator [see equation (1)] and u = T{T} + S {S}, if and only if L1T = 0 and L2S = 0.GP2 OrthogonalityGP3 Closure: ∇ × S{S} is a T field.Two choices of T and S fields are considered: type I fields with potentials T and S, which may depend on ξ1, ξ2 and ξ3, and type II fields with ξ1-independent potentials. It is shown that properties GP1–GP3 only hold for type I fields in spherical and cylindrical coordinate systems, and for type II fields in azimuthal and cylindrical coordinate systems with axisymmetric and two-dimensional potentials, respectively. Analogues of GP1 for the vector wave and diffusion equations, and the Navier equation of linear elasticity, are also only true in the same four cases. Generalisations of type I and II T and S fields to arbitrary coordinate systems are indicated.