ABSTRACT
Plasmatic homocysteine concentration depends mostly on 5,10 methylene tetrahydrofolate reductase (MTHFR) polymorphisms, a key enzyme in folate metabolism. The most common point mutation C677T is associated to cardiovascular and neurological pathologies; its ethnic repartition is quite heterogenic. In the present study, we proposed to describe the genotypic and allelic frequencies of C677T polymorphism and its influence on plasmatic homocysteine level in a healthy Algerian population. The investigation was turned on 100 apparently healthy voluntary subjects. Homocysteine concentration was determined using an immunoassay by fluorescence polarisation on IMx. Genotypes were determined by RT-PCR (Light cycle 480). Mean homocysteine concentration value was 14,69 +/- 7,30 micromol/L. 41% of people sample show a moderate hyperhomocysteinemia (>15 micromol/L). For the MTHFR C677T, estimated frequency of the allele T in the 100 people sample was about 35,5% with genotypic frequency of 6%. Plasmatic homocysteine is significantly higher in people carrying allele T: (CC vs CT: 11,8 +/- 2,97 micromol/L vs 15,47 +/- 6,74 micromol/L, p = 0,0004); (CC vs TT: 11,8 +/- 2,97 micromol/L vs 30,05 +/- 13,35 micromol/L, p = 0,01) and (CT vs TT: 15,47 +/- 6,74 micromol/L vs 30,05 +/- 13,35 micromol/L, p = 0,021). Our study shows an intermediate allelic frequency that joins the North-South world gradient and a high hyperhomocysteinemia prevalence. C677T polymorphism of MTHFR seems playing a predominant role in the moderate hyperhomocyteinemia. These two observations should be taken into consideration in the evaluation of morbid and/or lethal pathologies predisposition in the Algerian population.
Subject(s)
Homocysteine/genetics , Hyperhomocysteinemia/genetics , Methylenetetrahydrofolate Reductase (NADPH2)/genetics , Point Mutation , Polymorphism, Genetic , Adolescent , Adult , Algeria , Data Interpretation, Statistical , Female , Fluorescence Polarization Immunoassay , Gene Frequency , Genotype , Homocysteine/blood , Humans , MaleABSTRACT
1. The glycopeptide carrying the glycosaminoglycan chain of the urinary trypsin inhibitor (immunologically and structurally related to inter-alpha-trypsin inhibitor) was isolated. 2. The data from amino acid composition and part sequencing of this glycopeptide unambiguously demonstrate that the glycosaminoglycan is covalently linked to serine-10 of the peptide chain of UTI.
Subject(s)
Chondroitin Sulfates/analysis , Glycopeptides/chemistry , Glycoproteins/chemistry , Trypsin Inhibitors/chemistry , Amino Acid Sequence , Animals , Glycopeptides/isolation & purification , Molecular Sequence Data , Serine/chemistry , Serine Endopeptidases/metabolismABSTRACT
cDNA studies have suggested that inter-alpha-trypsin inhibitor (ITI) is a complex of several different peptide chains; the sequence of the inhibitory part of ITI is in excellent agreement with that of the urinary trypsin inhibitor (UTI). The present report demonstrates that a compound immunologically related to UTI is released by digestion with porcine pancreatic elastase or human leucocyte elastase. Since UTI has been shown to be a proteoglycan, ITI has been treated by chondroitinase. In these conditions, ITI is dissociated and gives rise to two heavy chains (78 and 85 kDa) and one light chain (26 kDa) immunologically related to UTI and which in PAGE moves close to UTIc (produced by chondroitinase treatment of UTI). We suggest that ITI is a non-covalent complex comprising two heavy chains and one light chain immunologically related to UTI and which is also a proteoglycan.
Subject(s)
Alpha-Globulins/metabolism , Proteoglycans/physiology , Trypsin Inhibitors/metabolism , Blotting, Western , Chondroitin Lyases , Electrophoresis, Polyacrylamide Gel , Glycosaminoglycans , Humans , Hydrolysis , Immunoelectrophoresis , Indicators and Reagents , Pancreatic Elastase , Trypsin Inhibitors/urineABSTRACT
SDS-polyacrylamide gel electrophoresis and immunoblot were applied to analysis of plasma proteins immunologically related to inter-alpha-trypsin inhibitor (ITI). In this system, anti-ITI sera were able to identify ITI and other components with an Mr near 120 kDa which would be degradation products of ITI by limited proteolysis. An anti-UTI (urinary trypsin-inhibitor) serum could detect, beside these derivatives, two minor components (Mr values near 90 and 60 kDa). Analysis of perchloric acid supernatants of plasma samples, using the same technic, induced visualization of a new component, similar to urinary trypsin inhibitor which could not be detected by direct analysis. This one was also characterized in a higher content in pathological samples (renal failure and infectious diseases).
