ABSTRACT
Matrix metalloproteinases (MMPs) are a family of zinc-dependent metalloendopeptidases that degrades extracellular matrix (ECM) components. MMPs are associated with venous wall remodelling, proliferation, migration, phenotypic and functional transformation of vascular smooth muscle cells and ECM organization under the physiological and pathophysiological conditions. We investigated possible association of genetic promoter polymorphisms of MMP2 (rs243866), MMP8 (rs11225395), MMP9 (rs3918242) and TIMP2 (rs8179090) to varicose veins development in the Slovak population. Genomic DNA from 276 Slovak individuals (138 cases, 138 controls) was genotyped for selected SNPs (rs243866, rs11225395, rs3918242 and rs8179090) using the PCR-RFLP analysis. The data were analysed by chi-squared (chi2) test, logistic regression, and Mann-Whitney test. The risk of varicose veins development was evaluated in dominant, codominant and recessive genetic models. The statistical evaluation of selected polymorphisms in patients in all three genetic models has not shown a significant risk of varicose veins development. Our study has not shown the association between selected polymorphisms and increased risk of varicose veins development in Slovak population. More evidence with broaden sample size is needed.
Subject(s)
Matrix Metalloproteinases/genetics , Tissue Inhibitor of Metalloproteinase-2/genetics , Varicose Veins/genetics , Adolescent , Adult , Aged , Case-Control Studies , Female , Genetic Predisposition to Disease , Genotype , Humans , Male , Matrix Metalloproteinase 2/genetics , Matrix Metalloproteinase 8/genetics , Matrix Metalloproteinase 9/genetics , Middle Aged , Pilot Projects , Polymorphism, Single Nucleotide , Promoter Regions, Genetic , Slovakia/epidemiology , Varicose Veins/epidemiology , Varicose Veins/pathology , Young AdultABSTRACT
Our current knowledge of antigenic variability of the bovine respiratory syncytial virus (BRSV) is quite limited and is mainly dependent on the use of monoclonal antibodies (mAb). In this study, we present not only analysis of the antigenic, but also of the genetic variability of BRSV. Using a panel of BRSV-specific mAb we distinguished five main reactivity patterns, three of which corresponded to the previously established subgroups A, B and AB. A single viral strain yielded the fourth pattern, while four viral strains did not react with any of the used mAbs forming the fifth pattern. To investigate the genetic basis for the antigenic heterogeneity of the BRS virus G protein, DNA of 11 BRSV isolates was directly sequenced. The comparison of the obtained nucleotide or amino acid sequences to those BRSV strains present in the GenBank revealed 88.1-99.4% and 77.7-98.4% similarity, respectively. These results supported the previously stated suggestion to type BRSV isolates according to their genetic relationship. In order to introduce a rapid and simple method to study the genetic variability of BRSV, we utilized the restriction enzyme analysis of RT-PCR products derived from mRNAs corresponding to the most variable region of the BRSV glycoprotein G ectodomain. Using this restriction enzyme analysis we were able to identify genetic variability among BRSV isolates. The detected non-synonymous mutations led frequently to a change in digestion pattern and were predominantly located in two mucin-like regions of the G protein gene. A correlation has been found between grouping of isolates in the phylogenetic tree and their restriction patterns clustering together isolates with the same restriction profiles. However, viruses placed distant in the tree sharing the same restriction patterns were detected supposing that phylogenetic analysis should be necessary for BRSV typing. Thus, we propose to use DNA restriction polymorphism for a rapid detection of genetic variants among BRSV isolates circulating in cattle population and as a preliminary tool for their typing.
Subject(s)
Genetic Variation , Respiratory Syncytial Virus, Bovine/genetics , Restriction Mapping/methods , Viral Proteins/genetics , Amino Acid Sequence , Antibodies, Monoclonal , Antibodies, Viral , Antigens, Viral/genetics , Molecular Sequence Data , Phylogeny , Reverse Transcriptase Polymerase Chain Reaction , Sequence Alignment , Sequence Homology, Amino AcidABSTRACT
In this study we showed a high degree of genetic homogeneity among recently (2002-2003) circulating Bovine respiratory syncytial virus (BRSV) strains in cattle population in the Czech Republic. These strains are in a phylogenetic tree more closely related to the Danish strains from 1995 than to the Czech strain VS97 from 1997 that shares the highest similarity with the French strain F1 and the Belgian strain P10. From the sequence analysis we deduce that the revealed high diversity between BRSV strains from 2002-2003 and those from 1997, at both nucleotide (0-11.4%) and amino acid (0-21%) level, is more likely due to distinct sources of the virus strains than to the sequence evolution.
Subject(s)
Cattle Diseases/virology , Respiratory Syncytial Virus Infections/veterinary , Respiratory Syncytial Virus, Bovine/classification , Amino Acid Sequence , Animals , Cattle , Cattle Diseases/epidemiology , Czech Republic/epidemiology , Molecular Sequence Data , Phylogeny , Respiratory Syncytial Virus Infections/epidemiology , Respiratory Syncytial Virus Infections/virology , Respiratory Syncytial Virus, Bovine/genetics , Respiratory Syncytial Virus, Bovine/immunology , Viral Envelope ProteinsABSTRACT
RNA of Bovine respiratory syncytial virus (BRSV) was found in peripheral leukocytes and nasal mucosa of infected cows by nested reverse transcription-polymerase chain reaction (nRT-PCR). We suppose that this finding obtained in the convalescent phase of infection indicates possible persistence of the virus in cells of the immune system.
