Isoenzyme characterization of trichomonas vaginalis isolated from hiv patients in fars and kerman, Southeast Iran

Background: Trichomonas vaginalis is an anaerobic flagellated protozoan which is responsible for human urogenital infections. Several zymodemes of T. vaginalis have been reported from various parts of the worlds on the basis of isoenzyme patterns. This study was conducted to characterize the isolated organisms of T. vaginalis from HIV patients using isoenzyme electrophoresis in Fars and Kerman provinces, southeast Iran. Methods: Eighteenmass cultivated isolates of T. vaginalis in themodified TYI-S-33medium were analyzed using isoenzyme electrophoresis. Polyacrylamide gel electrophoresis (PAGE) of five different enzyme systems were used to characterize T. vaginalis isolates: (i) Glucose-6-phosphate dehydrogenase (G6PD), (ii) Glucose phosphate isomerase (GPI), (iii) Malate dehydrogenase (MDH), (iv) Malic enzyme (ME), and (v) Phosphoglucomutase (PGM). Results: MDH, GPI, PGM, and ME enzyme systems showed a homogeneity and detected an identical enzyme pattern in all isolates. Meanwhile, G6PD revealed two different enzyme patterns. The isoenzyme electrophoretic profiles divided 18 T. vaginalis isolates into two zymodemes. Zymodeme 1 contained Shiraz isolates and zymodeme 2 contained Kerman isolates. Conclusion: The polymorphism of Iranian human isolates of T. vaginalis could be assessed by biochemical study using appropriate enzyme systems. Isoenzyme analysis is a promising method for the characterizationof T. vaginalis. New molecular studies with increased number of enzyme loci and genetic markers are suggested to classify more zymodemes of Trichomonas in Iran

Isoenzyme profiles and phylogenetic analysis of Giardia duodenalis isolates from Iranian patients

The main objective of this study was to characterize the Giardia duodenalis isolates from Iranian patients in Fars Province, south of Iran by biochemical and molecular methods. Fifteen mass cultivated of G. duodenalis isolates in modified TYI-S-33 medium were analyzed using isoenzyme electrophoresis and PCR genotyping. Polyacrylamide gel electrophoresis (PAGE) of five different enzyme systems was used to characterize isolates: (i) glucose-6-phosphate dehydrogenase, (ii) glucose phosphate isomerase, (iii) malate dehydrogenase, (iv) malic enzyme, and (v) phosphoglucomutase. As well, a fragment of the SSU-rDNA (292 bp) gene was amplified by PCR using the primers RH11 and RH4. The sequencing of the PCR products and phylogenetic tree were performed. The isoenzyme electrophoretic profiles divided fifteen G. duodenalis isolates into four zymodemes. G6PD, GPI, MDH, ME, and PGM enzyme systems showed 1, 2, 2, 3, and 3 enzyme pattern, respectively. G6PD isoenzyme pattern had the most homogeneity, while isoenzyme patterns of ME and PGM had the most heterogeneity in our study. Genotyping results indicated that the zymodemes 1–4 were categorized in assemblage A based on the SSU-rDNA gene. Phylogenetic analysis showed that all four zymodemes were distributed within the cluster of assemblage A. Our results indicated that both isoenzyme and DNA analyses were useful to characterize the isolates of Giardia and distinguishing various zymodemes and assemblages. It could be suggested that the genetic diversity among isoenzymes profiles of G. duodenalis may explain the variable clinical manifestations, pathogenicity, host response, drug susceptibility, and treatment efficacy of human giardiasis

Efficacy of prophylaxis and genotype-phenotype correlation in patients with severe Factor X deficiency in Iran

We aimed to evaluate the effect of regular prophylaxis with a Factor X (FX) concentrate for patients with severe FXD in Iran and to assess the correlation of the genotype and phenotype in these patients. Ten patients with severe FXD (FX activity <1%) were enrolled and characterized during 2010-2011. Prophylaxis with 20\u2003IU FX P Behring per kg body weight was administered once a week. FX levels, were monitored at baseline, 15 and 30\u2003min, 1, 3, 6, 12, 24, 48, 72 and 96\u2003h after starting prophylaxis. All patients were followed for 1\u2003year. The mean age of the patients was 15\u2003\ub1\u20037.8\u2003years (age range of: 6-27\u2003years). One patient had anaphylactic reaction after the first infusion, and the treatment was stopped. During one-year follow-up after starting prophylaxis, no bleeding symptoms occurred in any patient who tolerated and remained on the prophylaxis programme and all of them had a FX level of 1% or above. The maximum level of FX activity has been observed at 15\u2003min after starting prophylaxis. A level of 1.5-3.5% was detected after 96\u2003h. Homozygous mutations p.Arg40Thr (Arg-1Thr), p.Gly51Arg and p.Glu69Lys were detected in patients with intracranial haemorrhage. In our patients, significant decrease in symptoms without any complication after administration of FX, was demonstrated in all except one patient who had an anaphylactic reaction. It seems that the dose of 20\u2003IU\u2003kg(-1) could be probably the best choice for patients with severe FXD, who require regular prophylaxi