Advanced vacuolation indicates propagation of various salmonid alphavirus type 2 isolates in Acholeplasma-infected BF-2 cells

2020 ◽  
Vol 139 ◽  
pp. 189-197
Author(s):  
O Schachner ◽  
N Dinhopl ◽  
A Friedl ◽  
H Soliman ◽  
M El-Matbouli
Keyword(s):  
Rainbow Trout ◽  
Brook Trout ◽  
Salvelinus Alpinus ◽  
Post Inoculation ◽  
Specific Pcr ◽  
Pcr Product ◽  
16S Rna ◽  
Infected Cells ◽  
Nsp3 Gene

During previous routine inspections of bluegill fry (BF-2) and rainbow trout gonad (RTG-2) cells incubated with organ samples from asymptomatic Arctic char Salvelinus alpinus, brook trout Salvelinus fontinalis, and rainbow trout Oncorhynchus mykiss, a distinctive, reproducible cytopathic effect (CPE) appeared. The striking CPE, involving progressive vacuolation turning into slowly proceeding pyknotic degeneration, was originally attributed exclusively to enhanced growth of Acholeplasma sp. However, at a recent re-examination of re-infected BF-2 cells using electron microscopy (EM), conventional PCR, and quantitative PCR (qPCR), a virus was also detected. Two days post inoculation (dpi), EM revealed characteristic virions inside cytoplasmic vacuoles and next to bacteria outside the cells. The nucleotide sequences of the viral nsP3 gene fragment obtained from supernatants of infected cells were 100% identical and representative for salmonid alphavirus type 2 (SAV 2). The 16S RNA gene (16S rDNA) fragment sequences of the Mollicutes-specific PCR product obtained from SAV-infected as well as virus-free BF-2 control cells were identical with Acholeplasma laidlawii. In addition, qPCR results indicated enhanced propagation of virus and bacteria increasing with vacuolation between 5 and 8 dpi. Advanced vacuolation can be regarded as a CPE of both SAV and A. laidlawii, suggesting a viral impact on the bacterial infection that turns a latent intracellular stage into an apparent degenerative condition.

Analysis of the Potential Association of Drug-Metabolizing Enzymes CYP2C9*3 and CYP2C19*3 Gene Variations With Type 2 Diabetes: A Case-Control Study

2020 ◽  
Vol 21 (14) ◽  
pp. 1152-1160
Author(s):  
Imadeldin Elfaki ◽  
Rashid Mir ◽  
Faisel Mohammed Abu-Duhier ◽  
Chandan Kumar Jha ◽  
Adel Ibrahim Ahmad Al-Alawy ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Antiplatelet Drug ◽  
Drug Metabolizing Enzymes ◽  
Genotype Distribution ◽  
Nucleotide Polymorphisms ◽  
Metabolizing Enzymes ◽  
Specific Pcr ◽  
Increased Risk ◽  
Different Populations

Background:: Cytochrome P450s (CYPs) are drug-metabolizing enzymes catalyzing the metabolism of about 75% of drug in clinical use. CYP2C9 represents 20% CYP proteins in liver cells and is a crucial member of CYPs superfamily. CYP2C19 metabolizes very important drugs such as antiulcer drug omeprazole, the antiplatelet drug clopidogrel and anticonvulsant mephenytoin. Single nucleotide polymorphisms (SNPs) of CYP genes have been associated with unexpected drug reactions and diseases in different populations. Objective:: We examined the associations of CYP2C9*3 (rs1057910) and CYP2C19*3 (rs4986893) with T2D in Saudi population. Methods:: We used the allele-specific PCR (AS-PCR) and DNA sequencing in 111 cases and 104 controls for rs1057910, and in 119 cases and 110 controls for rs4986893. Results:: It is indicated that the genotype distribution of rs1057910 in cases and controls were not significantly different (P=0.0001). The genotypes of rs1057910 were not associated with type 2 diabetes (T2D) (P>0.05). Whereas the genotype distribution of rs4986893 in cases and controls was significantly different (P=0.049). The AA genotype of rs4986893 may be associated in increased risk to T2D with OR=17.25 (2.06-143.8), RR=6.14(0.96-39.20), P=0.008. Conclusion:: The CYP2C9*3 (rs1057910) may not be associated with T2D, while CYP2C19*3 (rs4986893) is probably associated with T2D. These findings need to be validated in follow-up studies with larger sample sizes and different populations.

scholarly journals Reduced sphingosine kinase 1 activity in dengue virus type-2 infected cells can be mediated by the 3′ untranslated region of dengue virus type-2 RNA

2013 ◽  
Vol 94 (11) ◽  
pp. 2437-2448 ◽  
Author(s):  
J. M. Carr ◽  
T. Kua ◽  
J. N. Clarke ◽  
J. K Calvert ◽  
J. R. Zebol ◽  
...  
Keyword(s):  
Cell Death ◽  
Dengue Virus ◽  
Virus Type ◽  
Elongation Factor ◽  
Sphingosine Kinase ◽  
Sphingosine Kinase 1 ◽  
Diarrhea Virus ◽  
Infected Cells ◽  
Dengue Virus Type 2

Sphingosine kinase 1 (SphK1) is a lipid kinase with important roles including regulation of cell survival. We have previously shown reduced SphK1 activity in cells with an established dengue virus type-2 (DENV-2) infection. In this study, we examined the effect of alterations in SphK1 activity on DENV-2 replication and cell death and determined the mechanisms of the reduction in SphK1 activity. Chemical inhibition or overexpression of SphK1 after established DENV-2 infection had no effect on infectious DENV-2 production, although inhibition of SphK1 resulted in enhanced DENV-2-induced cell death. Reduced SphK1 activity was observed in multiple cell types, regardless of the ability of DENV-2 infection to be cytopathic, and was mediated by a post-translational mechanism. Unlike bovine viral diarrhea virus, where SphK1 activity is decreased by the NS3 protein, SphK1 activity was not affected by DENV-2 NS3 but, instead, was reduced by expression of the terminal 396 bases of the 3′ UTR of DENV-2 RNA. We have previously shown that eukaryotic elongation factor 1A (eEF1A) is a direct activator of SphK1 and here DENV-2 RNA co-localized and co-precipitated with eEF1A from infected cells. We propose that the reduction in SphK1 activity late in DENV-2-infected cells is a consequence of DENV-2 out-competing SphK1 for eEF1A binding and hijacking cellular eEF1A for its own replication strategy, rather than a specific host or virus-induced change in SphK1 to modulate viral replication. Nonetheless, reduced SphK1 activity may have important consequences for survival or death of the infected cell.

Food Specialization By Individual Trout

1972 ◽  
Vol 29 (11) ◽  
pp. 1615-1624 ◽  
Author(s):  
James E. Bryan ◽  
P. A. Larkin
Keyword(s):  
Rainbow Trout ◽  
Brook Trout ◽  
Salvelinus Fontinalis ◽  
Stomach Contents ◽  
Cutthroat Trout ◽  
Salmo Gairdneri ◽  
Potential Prey ◽  
Repeated Observation ◽  
Size Growth ◽  
Salmo Clarki

Analyses of stomach contents showed that the kinds of prey eaten by brook trout (Salvelinus fontinalis), cutthroat trout (Salmo clarki), and rainbow trout (Salmo gairdneri) were seldom distributed at random among the individuals. Repeated observation of food eaten by individuals in a stream and ponds showed that prey types were eaten in proportions which were characteristic for an individual.Specialization occurred on several different kinds of prey. Although the degree of specialization was higher during shorter intervals, the data suggested that some specialization persisted for half a year. There were no striking correlations between degree of specialization and other individual properties such as size, growth rate, weight of food, number of food items, previous specialization, or area of recapture.In addition to the observations on trout in relatively undisturbed habitats, a field experiment was conducted using laboratory-reared rainbow trout held in small ponds. The food of each trout in the experiment was sampled repeatedly. In analysis of variance, interaction among the individuals and kinds of prey eaten showed that food specialization occurred. Both the absolute and relative abundance of potential prey were constant during the experiment.

scholarly journals Herpes Simplex Virus Type 2 Mucin-Like Glycoprotein mgG Promotes Virus Release from the Surface of Infected Cells

Viruses ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 887
Author(s):  
Edward Trybala ◽  
Nadia Peerboom ◽  
Beata Adamiak ◽  
Malgorzata Krzyzowska ◽  
Jan-Åke Liljeqvist ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Virus Type ◽  
Herpes Simplex Virus Type ◽  
Culture Medium ◽  
Glycoprotein G ◽  
Infected Cells ◽  
Virus Interaction ◽  
Simplex Virus

The contribution of virus components to liberation of herpes simplex virus type 2 (HSV-2) progeny virions from the surface of infected cells is poorly understood. We report that the HSV-2 mutant deficient in the expression of a mucin-like membrane-associated glycoprotein G (mgG) exhibited defect in the release of progeny virions from infected cells manifested by ~2 orders of magnitude decreased amount of infectious virus in a culture medium as compared to native HSV-2. Electron microscopy revealed that the mgG deficient virions were produced in infected cells and present at the cell surface. These virions could be forcibly liberated to a nearly native HSV-2 level by the treatment of cells with glycosaminoglycan (GAG)-mimicking oligosaccharides. Comparative assessment of the interaction of mutant and native virions with surface-immobilized chondroitin sulfate GAG chains revealed that while the mutant virions associated with GAGs ~fourfold more extensively, the lateral mobility of bound virions was much poorer than that of native virions. These data indicate that the mgG of HSV-2 balances the virus interaction with GAG chains, a feature critical to prevent trapping of the progeny virions at the surface of infected cells.

Aeromonas salmonicida: Relationship between Extracellular Growth Products and Isolate Virulence

1981 ◽  
Vol 38 (11) ◽  
pp. 1322-1326 ◽  
Author(s):  
Rocco C. Cipriano ◽  
Billy R. Griffin ◽  
Burton C. Lidgerding
Keyword(s):  
Rainbow Trout ◽  
Brook Trout ◽  
Salvelinus Fontinalis ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Aeromonas Salmonicida ◽  
Salmo Gairdneri ◽  
Lesion Development ◽  
Culture Supernatants ◽  
Crude Material

Extracellular growth products, extracted from culture supernatants of Aeromonas salmonicida by precipitation with ammonium sulfate and ethanol, were resolved into four fractions by ion-exchange chromatography. Although one of these (fraction II) possessed leukocytolytic activity, virulence of the pathogen was not associated with leukocytotoxicity. A positive correlation was established, however, between virulence and the toxicity of extracted material to cultured rainbow trout gonad (RTG-2) cells. The crude material and fractions II and III from virulent isolates were more toxic to RTG-2 cells than were similar preparations from avirulent isolates. Preparations from virulent isolates caused hemorrhaging, lesion development, and mortality when injected intramuscularly into brook trout (Salvelinus fontinalis) and Atlantic salmon (Salmo solar). Rainbow trout (Salmo gairdneri) were more resistant and lesions could be produced only with fraction II. Fraction II also possessed proteolytic activity.Key words: Aeromonas salmonicida, furunculosis, pathogenicity, salmonids, extracellular products, cytotoxicity, leukocytolysis, proteolysis, fishes

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