ABSTRACT
Objective@#This study aimed to investigate the effects of @*Methods@#In this study, 0.1% DMG was supplemented in 20% casein diets that were either folate-sufficient (20C) or folate-deficient (20CFD). Blood and liver of rats were subjected to assays of Hcy and its metabolites. Hcy and its related metabolite concentrations were determined using a liquid chromatographic system.@*Results@#Folate deprivation significantly increased pHcy concentration in rats fed 20C diet (from 14.19 ± 0.39 μmol/L to 28.49 ± 0.50 μmol/L; @*Conclusion@#DMG supplementation exhibited hypohomocysteinemic effects under folate-sufficient conditions. By contrast, the combination of folate deficiency and DMG supplementation has deleterious effect on pHcy concentration.
Subject(s)
Animals , Male , Rats , Biomarkers/metabolism , Chromatography, Liquid , Diet , Dietary Supplements , Folic Acid Deficiency/metabolism , Homocysteine/metabolism , Liver/metabolism , Random Allocation , Rats, Wistar , Sarcosine/metabolismABSTRACT
<p><b>OBJECTIVE</b>Human cytomegalovirus (HCMV) displays genetic polymorphisms. Nineteen open reading frames (ORFs, UL133-UL151) found in the Toledo strain of HCMV and other low-passage clinical isolates may be essential for viral infection. This study aimed to analyze the polymorphism of HCMV UL134 gene in clinical isolates and explore the relationship between the polymorphism and HCMV infection.</p><p><b>METHODS</b>PCR was performed to amplify entire UL134 region in 32 clinical isolates, which had been proven as HCMV-DNA positive by FQ-PCR. PCR products were sequenced.</p><p><b>RESULTS</b>All of the 32 isolates were amplified and sequenced successfully. HCMV UL134 gene was highly conserved in the clinical isolates. UL134 ORF and its predicted protein in the clinical strains displayed 96.4%-98.3% nucleotide identity and 92.7%-94.9% amino acid identity respectively compared to those in the Toledo strain. A new posttranslational modification site, sulfationcamp (SUL) site, was found in UL134 protein of all of the clinical isolates except 35j.</p><p><b>CONCLUSIONS</b>HCMV UL134 gene in clinical isolates was highly conserved. No substantial relation was found between UL134 gene and HCMV infectious diseases.</p>
Subject(s)
Humans , Cytomegalovirus , Genetics , Cytomegalovirus Infections , Virology , Genes, Viral , Open Reading Frames , Polymerase Chain Reaction , Polymorphism, Genetic , Viral Proteins , GeneticsABSTRACT
<p><b>BACKGROUND</b>Human cytomegalovirus (HCMV) infection is an important infectious agent that results in neonatal disease and congenital deformity. HCMV infection may affect in many organs. The different symptoms and tissue tropism of HCMV infection perhaps resulted from the genetic polymorphism of HCMV. HCMV UL144 open reading frames encode a homologue of the tumor necrosis factor receptor. It seems important to study the strain-specific variability of UL144 sequence in low-passage clinical isolates and to discuss if the variability related to the clinical HCMV infection.</p><p><b>METHODS</b>HCMV-UL144 gene was amplified by PCR assay in 65 low-passage clinical isolates and urine from 7 healthy children who were HCMV-DNA positive by quantitative PCR. All the positive PCR products were analyzed by Heteroduplex mobility assay and single-stranded conformation polymorphism (HMA-SSCP) and 32 of them were sequenced.</p><p><b>RESULTS</b>Fifty-five isolates and 5 urine specimens were HCMV-UL144 positive by PCR. Sequencing and HMA-SSCP analysis showed that significant strain-specific variability was present in the UL144 ORFs. Comparing UL144 sequences and the corresponding symptoms showed that genotype 2 did not exist in megacolon isolates. And genotype 1 and 3 were the major types among microcephaly and jaundice isolates respectively.</p><p><b>CONCLUSION</b>HCMV-UL144 existed in almost all the low passage isolates. HMA-SSCP assay is an easy and effective method to detect the genetype of HCMV-UL144 sequence. The characteristic of sequences in different isolates showed that UL144 gene may play an important role in HCMV infection.</p>
Subject(s)
Humans , Infant , Infant, Newborn , Cytomegalovirus , Classification , Genetics , Cytomegalovirus Infections , Virology , Genotype , Membrane Glycoproteins , Genetics , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Sequence Analysis, DNA , Viral Proteins , GeneticsABSTRACT
<p><b>BACKGROUND</b>To study the polymorphism of human cytomegalovirus US28 gene in children and investigate the relationship between the polymorphism and pathogenesis.</p><p><b>METHODS</b>The FQ-PCR was carried out to determine the DNA quantity of clinical isolate and then the segmental PCR and HMA-SSCP were performed to test the mutation of US28 gene. The typical isolates from different diseases were selected to clone and sequence, then the results were analyzed.</p><p><b>RESULTS</b>The nucleic acid mutation is frequent among the sequence of US28, those mutations focus on the two ends of US28, but most of them are sense mutation. The important functional groups of US28 are highly conserved. The amino acid mutation of some isolates resulted in the change of secondary structure, but the phylogenetic tree analysis did not show any clear association between the pathogenesis and the distribution of clinical isolates. The comparison of US28 sequences from AIDS patients with the sequences from children in our study showed that both sequences have their own specific high mutation points.</p><p><b>CONCLUSION</b>There is polymorphism among the HCMV-US28 gene of clinical isolates from children. There observed no clear relationship was between the pathogenesis and the distribution of clinical isolates.</p>
Subject(s)
Child , Humans , Base Sequence , Congenital Abnormalities , Virology , Cytomegalovirus , Genetics , Cytomegalovirus Infections , Virology , Genotype , Molecular Sequence Data , Mutation , Polymerase Chain Reaction , Polymorphism, Genetic , Polymorphism, Single-Stranded Conformational , Receptors, Chemokine , Genetics , Sequence Analysis, DNA , Viral Proteins , GeneticsABSTRACT
<p><b>BACKGROUND</b>To investigate the polymorphism of human cytomegalovirus UL148 gene in low passage clinical isolates and to study the relationship between the polymorphism and different pathogenesis of congenital HCMV infection.</p><p><b>METHODS</b>PCR was performed to amplify the entire HCMV UL148 gene region of 38 clinical isolates, which had been proven containing detectable HCMV-DNA by using FQ-PCR.PCR amplification products were sequenced directly and the sequence data were analysed.</p><p><b>RESULTS</b>Seventeen of 38 isolates were amplified successfully. By comparison with Toledo sequence, the length of UL148 ORFs in all 17 clinical isolates was similar to that of Toledo. Amino acid variability rate of UL148 protein was 0.3%-2.3%. There were additional or deleted sites of posttranslational modification of UL148 protein in all clinical isolates.</p><p><b>CONCLUSION</b>All DNA and deduced amino acid sequences of UL148 gene shared great similarity among HCMV clinical strains regardless of their polymorphism.</p>