Overexpression of Ubiquitin and Amino Acid Permease Genes in Association with Antimony Resistance in Leishmania tropica Field Isolates

The mainstay therapy against leishmaniasis is still pentavalent antimonial drugs; however, the rate of antimony resistance is increasing in endemic regions such as Iran. Understanding the molecular basis of resistance to antimonials could be helpful to improve treatment strategies. This study aimed to recognize genes involved in antimony resistance of Leishmania tropica field isolates. Sensitive and resistant L. tropica parasites were isolated from anthroponotic cutaneous leishmaniasis patients and drug susceptibility of parasites to meglumine antimoniate (Glucantime(R)) was confirmed using in vitro assay. Then, complementary DNA-amplified fragment length polymorphism (cDNA-AFLP) and real-time reverse transcriptase-PCR (RT-PCR) approaches were utilized on mRNAs from resistant and sensitive L. tropica isolates. We identified 2 known genes, ubiquitin implicated in protein degradation and amino acid permease (AAP3) involved in arginine uptake. Also, we identified 1 gene encoding hypothetical protein. Real-time RT-PCR revealed a significant upregulation of ubiquitin (2.54-fold), and AAP3 (2.86-fold) (P<0.05) in a resistant isolate compared to a sensitive one. Our results suggest that overexpression of ubiquitin and AAP3 could potentially implicated in natural antimony resistance.

[Study on the association of Toxocariasis with allergic rhinitis in individuals referred to Rasoul Akram Hospital, Tehran]

Toxocariasis is a common worldwide zoonotic parasite infection caused by the larvae of Toxocara catti and Toxocara canis. Allergic rhinitis is the most common chronic diseases in the upper respiratory tract. The main symptoms are sneezing, watery rhinorrhea, itching in the nose, eyes and palate. The purpose of this study was to investigate the association between toxocara seropositivity and allergic rhinitis compared with the control population. This cross-sectional study was carried out from September 2009 to February 2011 on 93 patients with allergic rhinitis and 87 control subjects. Confirmation of the diagnosis of allergic rhinitis was defined by history and positive epicutaneous prick test. Control subjects were healthy based on history and no signs of allergic rhinitis and other allergic diseases were seen. Blood and fecal samples were taken from both groups. Sera were separated, labeled and stored at -20°C until used. Stool samples were examined by a wet mount and formalin-ethyl acetate concentration technique. The diagnosis of toxocariasis was established by IgG anti Toxocara and IgE total by ELISA method. In case group [allergic rhinitis] from 93 patients, 50 patients [53.8%] were males and 43 [46.2%] were female. In the control group of 87 individuals studied, 56 [64.4%] were males and 31 [35.6%] were female. In cases and controls, 5 [5.4%] and 3 [3.4%] of sera were positive for IgG Toxocara, respectively. There was no statistical difference in Toxocara seropositivity in both groups [p =0.39]. It seems to be in contrast to worms and allergies several factors, including phase worm infections [acute and chronic], parasite load, parasite species and resistance genes are involved and this require further studies in different ages and populations

Partially purified fraction [PPF] antigen from adult fasciola gigantica for the serodiagnosis of human fasciolisis using dot - Elisa Technique

Human fascioliasis has been reported in many countries, including Iran. Various techniques have been evaluated for diagnosis of human fascioliasis using different antigens. We evaluated Fasciola gigantica partially purified fraction antigen [PPF] isolated from sheep's liver fluke for the diagnosis of human fascioliasis. Materials and Two hundred sixty-one sera were collected from 104 patients living in an area endemic for human fascioliasis, from 89 non-fascioliasis patients living in a non-endemic area, and from 68 healthy individuals. Micro-ELISA was used in the evaluation of the sensitivity and specificity of Dot-ELISA. With a 1:800 sera dilution as the cut-off titer, the sensitivity of the Dot-ELISA test in diagnosis of human fascioliasis was 94.23% and the specificity was 99.36%. Dot-ELISA using PPF antigen is a sensitive and specific method for diagnosis of human fascioliasis that is also rapid and inexpensive

Evaluation of a dot-ELISA for the serodiagnosis of human hydatid disease

A dot-ELISA utilizing antigen-B from sheep hydatid fluid, dotted onto nitrocellulose filter discs was developed for the rapid diagnosis of human hydatidosis. 1micro g of antigen per dot, serum dilution of 1:800, dilution conjugate of 1:1000 and 45 min incubation were found optimal. Thirty four patients infected with hydatidosis, 32 cases with other parasitic diseases and 36 healthy subjects were included in the assay and were examined using dot-ELISA to detect antibody against the aforementioned parasite antigen. Sensitivity, specificity and positive and negative predictive values of the assay were calculated as 97.1%, 98.5%, 97.1% and 98.5% respectively. No false positive reaction was observed when 36 sera healthy subjects were assayed. One case of cross-reactions was observed as for a serum infected with fasciolosis. It was concluded that dot-ELISA is rapid, antigen and serum conservative as well as encompasses great importance to confirm clinical diagnosis either in the laboratory or in the filed