[Comparison of the toxicity spermicidal effect of some gram negative bacteria's lipopolysocharides, including that from Chlamydia Trachomatis on Human Sperm]

Chlamydia trachomatis infection is one of the most common sexually transmitted diseases especially in western countries. While there is agreement on the manifestations and, in particular, the negative effects of Chlamydia infection on female fertility, the role of this organism in male fertility is still controversial. In addition to Chlamydia, some bacteria belong to the enterobacteriaceae family, including: E.coli, Klebsiella, and Serattia have been suggested in human sperm dysfunction. The purpose of the present study was to compare the in vitro toxic effects of lipopolysaccharides [LPSs] extracted from these bacteria on human sperm. In this laboratory study, 5x10[6] sperm/ml of prepared sperm using percoll gradient method were treated with 0.1, 1, 10, 25, and 50 micro g/ml of commercial LPSs from E.coli, Klebsiella, and Serattia and 0.1 micro g/ml of lab-made Chlamydia LPS at 37degree sign C in 5% CO[2] for 6 hours. After 6h incubation the sperm viability was measured using the HOS test. Commercial LPSs used in this study had no significant detrimental effects on human sperm at lower than 50 micro g/ml while Chlamydia LPS at 0.1 micro/ ml showed a significant toxic effects against sperm compare to control group [38.6 +/- 1, p<0.05]. Although, Chlamydia infections are almost clinically chronic and asymptomatic, our findings showed that the in vitro spermicidal activity of this bacterium is about 500 times more potent than LPS of E.coli, Klebsiella, and Serattia. These results and the findings from the other relevant studies confirm the key role of C.trachomatis as one of the infectious factors causing male infertility

Detection of common bacteria implicated in bovine mastitis in bulk tank milk by polymerase chain reaction

This study was conducted to detect the common bacteria implicated in bovine mastitis in bulk tank milk by polymerase chain reaction [PCR]. Forty-four milk samples from bulk tank milk were obtained and submitted to our laboratory. To detect Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Escherichia coli, Streptococcus uberis, and Streptococcus parauberis, two sets of universal primers were used. The PCR reaction was set up as described in previous literature. Using universal primers, PCR amplification results demonstrated 28 positive samples [63.6%] of 44. The percentages of positive samples for farms with production rates of < 3, 3-10, and 10 < tones were 48, 85, and 100%, respectively. In the present study, we concluded that using universal primers, a simplex PCR is able to detect common important bacteria implicated in bovine mastitis

[Evaluation of the stress effect on expression of BEST-5 gene in cultured rat hepatocytes and cloning of this gene]

Liver has important roles in body metabolic regulation and for this reason hepatocytes are used worldwide. Investigations showed that isolation of hepatocytes causes activation of stress related genes. The aim of this study was to study the stress related expression of BEST-5 following hepatocytes isolation and culture. The BEST-5 gene is cloned and analyzed for the first time from isolated and cultured rat hepatocytes. Very little is known about this gene and almost nothing is known about its function. RNA was isolated from hepatocytes after 3h culture and used for generation of PCR products corresponding to the BEST-5. cDNA generated was cloned into pCR[R]2.1 plasmid vector. Following transformation into TOPO10 oneshot [R]cells, the cells were grown in LB agar plates containing X-Gal and ampicillin, overnight at 37[oC]. To confirm that the plasmids contained inserts of the correct size, the vectors obtained from mini-preparations were digested with the desired restriction enzymes. Sequencing was performed for the gene. RT-PCR and Northern blotting analysis showed that BEST-5 mRNA is expressed, 3h after isolation and culture of primary hepatocytes [3h] BEST-5 mRNA was observed until 5h of culture and then there was no detectable band of BEST-5 at further time points. Comparison of expression of the level of mRNA of BEST-5, when data statistically were analyzed, showed a significant difference between the expression of BEST-5 mRNA expression at 3h with 0h, 24h, 35h and 48h of culture [P<0.001]. According to the results the stress induced by hepatocytes isolation and culture leads to the expression of Best-5 time-dependently

preparation and evaluation of reference Leishmanin from Leishmania major for use in man for diagnostic and experimental purposes

Cell-mediated immunity [CMI] plays an important role in resistance against leishmaniasis. Delayed-type hypersensitivity [DTH] reaction measured by skin testing is a practical method of CMI and is used as an aid to diagnosis and for epidemiological assessment of exposure to leishmanial infection. Skin testing in leishmaniasis, generally known as Montenegro or leishmanin test, requires a standard antigen. At present no uniform and standard leishmanin is available for skin testing in leishmaniasis. The present work describes the preparation and testing of an antigen from Leishmania major using standard conditions. Three dilutions of this antigen were tested in recovered individuals in an endemic area in Iran. The results obtained showed that leishmanin preparation exhibited high specificity and sensitivity, and strong potency