[Evalution of antileishmanial activity of the hydroalcoholic extract of medicago lupulina' leaves against leishmania major [MHOM/IR/75/ER] promastigotes by using MTT and microscopy method and GC/MS for hydroalcoholic extract of this plant' leaves]

Background: Leishmaniasis, has created enormous global health problem. Side effects, drug resistance and the lack of effective vaccines and to make the new compounds effective due to plant

Objective: The traditional medical plants such as black alfalfa can be a valuable source of new pharmaceutical agents against leishmaniasis

Methods: Alcoholic extracts were prepared by maceration method. L. major promastigotes [Leishmania major] in Schneider and then were cultured in RPMI- 1640. Then, using MTT [Methyl Thiazole Tetrazolium], the IC50 [Inhibitory Concentrations 50%] for extract and Glucantime was determined. MTT assay did for each sample, 3 times

Results: IC50 for alcoholic extract of alfalfa black against L. major promastigotes in vitro after 24, 48 and 72 hours, respectively 165, 98 and 45 micrograms per ml and for Glucantime also equal to 27, 12 and 8 mg l respectively. IC50 between Extract and Glucantime after 24, 48 and 72 hours there was a significant difference [P <0.05]. Morphological changes after challenge with meglumine and alcoholic extracts including cell shrinkage, round, dense cytoplasm and the cell was smaller. Presence of alkaloids and flavonoids in alcoholic extracts have been proved

Conclusion: As regards, plant extract had anti- leishmanial effects in vitro, further works are required to appraise the exact effect on Leishmania agent in animal models

[Molecular study of Clostridium difficile isolated from beef in Isfahan, Iran]

Background: clostridium difficile [C. difficile] infection is one of the most important diseases in healthcare facilities and community. Ribotypes 027 and 078 are known as hyper- virulent strain of C. difficile in molecular study. PCR-ribotyping is a suitable method to interpret the relation of C. difficile isolated from food and hospital

Objectives: in the present study, the clostridim difficile binary toxin [cdtB] and ribotype pattern evaluated in toxigenic C. difficle isolated from beef

Methods: detection of cdtB in 12 toxigenic C. difficile [encoding tcdA and tcdB gene] isolated from 100 beef samples was determined through PCR. Afterwards, PCR-ribotyping was performed to examine the ribotype patterns of C. difficile

Results: cdtB gene was not detected in any positive isolate. Ten different patterns were observed in 12 toxigenic isolates. No similarity existed in the ribotypes of our study with ribotypes 027 and 078

Conclusions: albeit ribotyp 027 and 078 were not found in our study, the isolation of toxigenic C. difficile with new ribotypes in Iran may indicate the probable hazard of this bacterium in public health. Comprehensive research about C. difficile in different food sources is recommended on a national level

[Identification of total and fecal coliforms and heterotrophic to Microbiological method and E.Coli O157:H7 to immunological, and real time PCR methods in Isfahan water treatment plant]

Total and Fecal coliforms [TC and FC], heterotrophic plate count [HPC], were counted by microbiological method and E.coli O157:H7 were detected through immunological and Real time PCR methods in water intake and all of units of Isfahan water treatment plant [IWTP]. The microbial profile including TC, FC, and HPC, were monitored and turbidity and total organic carbon were analyzed in 8 locations of water intake, and unit operation and processes of IWTP, including, inlet, sedimentation, ozonation, and filtration and finished water. Immunological method through anti-serum kits and molecular method of RT-PCR were used to detect E.coli O157:H7 in the 8 locations and also the sludge of the sedimentation basin and filters backwash water of IWTP. Survival of E.coli O157:H7 in sludge sample of sedimentation basin was indicated by formation of agglutination particles in immunological method and through indicator probes in the RT-PCR method. However, E.coli O157:H7 was not detected in water samples of other units of IWTP. The removal percent of TC, FC, and HPC were: 59.5, 49, and 54.8% in sedimentation basin; 66, 45.8, and 57% in ozonation;: 98.8, 98, and 78.8 in the filtration; and 96, 100, 91% in disinfection, respectively. This study approved the existence of the pathogenic coliform, E.coli O157:H7 in the sludge of sedimentation basin. Absent of E.coli O157:H7 in the finished water indicates that the existing units of IWTP could eliminate these pathogenic bacteria, before reaching the final units of the plant, including the filters and disinfection

[HLA-DR typing by PCR-SSP and comparison of serologic results with this method]

Introduction: Considering the role of HLA matching in transplant outcome, the quality of HLA-DR typing is clearly an important issue. In recent years serologic methods have been replaced with DNA based typing methods. The main objective of this study was to compare HLA-DR typing data obtained from existing serologic method with that obtained by the new PCR-SSP method

Material and Methods: Fifty-five peripheral blood samples were collected from randomly selected individuals who reffered to the transplantation laboratory of Isfahan in Aliasghar Hospital and was typed for HLA-DR antigens by both methods. HLA-DR typing by serologic method was performed using 30 different antisera and for PCR-SSP method specific primers were used for HLA-DRB1*01-10[except DR6, 8, 10] and also for HLA-DR52 and DR53. After DNA extraction 13 pairs of specific primers were used for each sample separately and PCR reaction was performed. In this study the third intron of DR locus was used as internal positive control. After PCR amplification, electrophoresis on 2% agarose gel was performed on reaction products, and after photography of gel, interpretation and comparison of results was performed

Results: The results of 31 samples [56.3%] correlated completely with serologic method. 12 samples [22%] had been assigned heterozygous in serology and homozygous in molecular typing, 7 samples [12.7%] were heterozygous in both methods but different in one allele, 2 samples [3.6%] were homozygous in serology and heterozygous in molecular typing, and also one sample [1.8%] was homozygous in both methods; that is serologically DR14 positive and moleculary DR11 positive. And finally 2 samples of 55 [3.6%] were not definable by in serologic method

Conclusion: The data obtained from the conventional serologic method has 43.7% discrepancy with PCR-SSP results, and this indicates the more error of serologic method and, in the other word, more accuracy of PCR-SSP technique for DR-typing