Actin Gene-Based Molecular Typing of Trichomonas vaginalis Clinical Isolates from the North of Iran.

AIM: The aim of the current study is to evaluate the prevalence of trichomoniasis in men and women in the north of Iran and to find genotypes in the positive clinical specimens based on T. vaginalis actin gene. MATERIALS AND METHODS: Women's genital (n = 500) and men's urine (n = 1500) samples were collected from the participants referred to clinics in Mazandaran Province, northern Iran, during 2006-2018. In addition, 1500 Pap smear specimens, archived in the Bu Ali Hospital, Sari City, Mazandaran Province, northern Iran, were examined. The specimens were examined based on parasitological methods, nested polymerase chain reaction (PCR), PCR-restriction fragment length polymorphism, and phylogenetic analysis. RESULTS: Overall, 17 (0.48%) of 3500 specimens were positive by PCR. Total prevalence was 0.55% (n = 2000) for women, of which 500 (1.4%; n = 7) specimens were collected freshly, and 1500 (0.26%; n = 4) were Pap smears. Moreover, six (0.4%) out of 1500 men urine specimens were positive. Overall, genotypes G, E, and I were detected with the prevalence of seven (0.2%), seven (0.2%), and three (0.08%), respectively. There was no significant statistical difference among the prevalence of the detected genotypes (P > 0.05). CONCLUSION: As a whole, the prevalence of trichomoniasis was low in the studied area in the north of Iran and, most importantly, the genotypes of E, G, and I were distributed among men and women in the province.

Hepcidin gene polymorphisms and iron overload in ß-thalassemia major patients refractory to iron chelating therapy.

BACKGROUND: ß Thalassemia is one of the most common groups of hereditary haemoglobinopathies. Affected people with thalassemia major are dependent on regular blood transfusion which on the long term leads to iron overload. Hepcidin is a peptide hormone and an important regulator of iron homeostasis, especially in thalassemia. Expression of this hormone is influenced by polymorphisms within the hepcidin gene, HAMP. Several studies emphasized the role of single nucleotide polymorphisms (SNPs) located in the promoter region of the gene. This study aimed to analyze the association between three SNPs in promoter of HAMP, c.-582A > G, c.-443C > T, and c.-153C > T, with iron overload in ß-thalassemia major patients. METHODS: A total of 102 samples from ß thalassemia major patients were collected. Genomic DNA was extracted and segments of DNA encompassing rs10421768 and rs142126068 were sequenced. Statistical analysis was performed by SPSS Statistics 23 using independent t test and Fisher's exact test. RESULTS: A total of 102 adult ß-thalassemia major patients were genotyped for three SNPs in the promoter region of HAMP gene by PCR and direct sequencing. Most of the patients (71.3%) were iron overloaded (based on plasma ferritin > 1000 ng/ml) in spite of receiving regular iron-chelating therapy. Our analysis revealed a statistically significant difference between the level of cardiac iron accumulation and c.-582A > G variant (p = 0.02). For c.-443C > T statistical analysis was on the edge of the significant relationship between the minor allele and serum ferritin (p = 0.058). All samples were homozygous for allele C of c.-153C > T. CONCLUSIONS: Despite chelating therapy, iron overload is still one of the main complications of thalassemia. Our findings and others emphasize the role of hepcidin -582A > G polymorphism as a key component of iron homeostasis in these patients.

Genetic diversity at the C-terminal domain of knob-associated histidine-rich protein (KAHRP) of Plasmodium falciparum isolates from Burundi, Eastern Africa.

The knob-associated histidine-rich protein (KAHRP) is an exported parasite protein and the major component of infected erythrocytes by Plasmodium falciparum. P. falciparum histidine-rich protein-1 (PfHRP-1) is docked by KAHRP, which this interaction plays a significant role in cytoadherence of the malaria protozoan to erythrocytes and pathogenicity. The most polymorphic region of the PfHRP-1 is the C-terminal of decapeptide repeat domain (region III). The main objective of this study was to explore the genetic diversity at the region III of KAHRP in P. falciparum isolates from Burundi. In the present study, the nested PCR was performed for the amplification of the coding gene (kahrp gene) for region III in 35 P. falciparum isolates from Burundi. The nested PCR products of seven randomly selected isolates were purified and then sequenced. As the result, three allelic forms (340 bp, 370 bp, and 400 bp) were seen at the C-terminal domain of kahrp gene. The existence of multiple alleles of the kahrp gene revealed the presence of different P. falciparum strains in Burundi. It is suggested that the results could be useful in designing and the improvement of targeted therapy agents for falciparum malaria.

Genetic polymorphism at the C-terminal domain (region III) of knob-associated histidine-rich protein (KAHRP) of Plasmodium falciparum in isolates from Iran.

The knob-associated histidine-rich protein (KAHRP) plays a major role in the virulence of Plasmodium falciparum and is one of the targets for molecular therapy. The primary structure of KAHRP of P. falciparum consists of three domains (regions I-III), of which the C-terminal domain (region III) is the most polymorphic segment of this protein. One of the main obstacles is genetic diversity in designing and developing of malaria control strategies such as molecular therapy and vaccines. The primary objective of the present study was to investigate and analyze the extent of genetic polymorphism at the region III of KAHRP of P. falciparum in isolates from Iran. A fragment of the kahrp gene spanning the C-terminal domain was amplified by nested PCR from 50 P. falciparum isolates collected from two malaria endemic areas of Iran during 2009 to August 2010 and sequenced. In this study, three allelic types were observed at the C-terminal domain of KAHRP on the basis of the molecular weight of nested PCR products and the obtained sequencing data. The presence of multiple alleles of the kahrp gene indicates that several P. falciparum strains exist in the malaria endemic areas of Iran. Our findings will be valuable in the design and the development of the molecular therapeutic reagents for falciparum malaria.