Study on Association of Recent and Past Chlamydophila pneumoniae Infection with Classic Multiple Sclerosis.

Aims: We conducted this study to determine if there is any correlation between Classical Multiple Sclerosis and Chlamydophilia pneumoniae infection by ELISA (IgM, IgG, IgA). Study Design: cross sectional study Place and Duration of Study: The present study was performed in the Department of Microbiology, Guilan University of Medical Sciences between April 2012 and April 2013 Methodology: Chlamydophila pneumoniae infection certified by ELISA in patients (n=46) and control (n=46) using commercial assays (anti- C. pneumoniae IgG, anti- C. pneumoniae IgM, and anti- C. pneumoniae IgA kits). Data were analyzed by using four statistical tests (Pearson chi square, Kendall's tau, and Spearman's rho). Results: Seropositivity to anti- C. pneumoniae IgG was seen less frequently in patients versus controls (69.0% versus 81.4%; P=0.187). Seropositivity to anti- C. pneumoniae IgA was also observed less frequently in patients than in controls (7.2 % versus 11.6%; P= 0.479).However anti- C. pneumoniae IgM antibodies were seen more often in classical multiple sclerosis patients than it was in controls ( 11.9% versus 2.3%; P= 0.085). Conclusion: We concluded that recent or past C. pneumoniae infection has no correlation in initiation or protection of CMS.

Detection of Pathogenic Legionella pneumophila in Tap Water and Water of Newborn Incubators in Hospitals of Guilan Province by PCR of mip Gene.

Aims: This study is designed to determine the frequency of Legionella pneumophila in cold and warm water as well as water containers of newborn incubators in Guilan province hospitals, Iran, using amplification of the macrophage infectivity protein gene (mip gene) by PCR. Study Design: Cross sectional study. Place and Duration of Study: The present study was performed in the Cellular and Molecular Research Center, Guilan University of Medical Sciences between June 2011 and July 2012 Methodology: Samples were collected directly in sterile containers, concentrated in centrifuge, transferred to yeast extract broth containing L- cysteine, Fe2+, Glycin and vancomycin and incubated for 3-4 days. DNA was extracted by using the boiling method and PCR was performed to search Legionella and mip gene using two pairs of primers. Contamination with other bacteria was evaluated in all negative samples using universal primers of 16S rRNA gene. Results: About 8.5% of the samples had L. pneumophila including 11% of the incubators and 5.8% of both hot and cold tap water. The mip gene was found in 2.8% of the samples. One third of the incubator and one half of the hot water habited L. pneumophila had the mip gene but it was not found in cold tap water samples. About 87.2% of the negative samples showed bacterial contamination as revealed by PCR with primers of 16S rRNA gene. Conclusions: This study indicates that in spite of using distilled water for incubators, L. pneumophila contamination is considerable and other bacterial contamination is very high. It may be related to the length of time that water remains in an incubator container which is a predisposing factor for both biofilm formation and the growth of water microflora. It seems that the high temperature of hot water system and the high rate of free residual chlorine in tap water system are the main causes of low rate of Legionella contamination but are ineffective on contamination with other bacteria.

[Antimicrobial effect of Ruta graveolens extract on pathogenic bacteria]

Ruta graveolens is a plant used in traditional medicine with variety of therapeutic effects. This study was done to evaluate the antimicrobial effect of hydro and alcoholic extracts of Ruta graveolens on 8 species of pathogenic bacteria. In this descriptive study, standard strains of Enterococcus faecalis, Staphilococcus aureus, Staphilococcus epidermis, Strptococcus pneumonia, Escherichia Coli, Klebsiella pneumonia, Salmonella typhi and Enterobacter Cloace were obtained from Iranian institute of scientific and industerial research center. The effect of hydro and hydroalcholic extracts of Ruta graveolens on the growth of bacteria was determined by disc diffusion and serial macro dilution methods. Minimum inhibitory concentration [MIC] for each extract was determined and the findings were compared with 8 common antibiotics including Co-trimoxazole, Oxycycline, Vancomycin, Erythromycin, Furazolidon, Tetracycline, amoxicillin / clavulanic acid and Ampicillin/sulbactam. Hydro and hydroalcoholic extracts of Ruta graveolens did not show any inhibitory effect on growth of the pathogenic bacteria even in the presence of high concentration [3mg/ml]. The extracts of arial organs of Ruta graveolens had no inhibitory effect on growth of the common human pathogenic bacteria

[ evaluation of melatonin effect on in-vitro development of mouse preimplantation embryos]

Melatonin promotes in-vitro embryo development in different species. This study studied the effects of melatonm on in-vitro mouse preimplantation embryo development. Two-cell embryos were obtained from oviduct of 6-8 weeks female NMRI mice 48 hours after administration of an intra-pentoneal injection of 5 lU/ml pregnant mares serum gonadotrophin and subsequent 5 ILJ/ml human chononic gonadotrophin [ip]. Embryos were cultured in T6 culture medium supplemented with different dosages of melatonin [0 [control group], 100x10[-6]M, 10 x10[-6]M, 1x10[-6], 100x10[-9]M and 10 x10[-9]M [1-5 treatment groups]]. With due attention to percent of embryos in different stages, the rate of development of embryos was assessed using of invert microscope. It is also compared to control group. The results showed that the rate of cleavage and development of mouse the embryos to blastocyst stage increased significantly in the development culture medium supplemented with 10 and 100 nM/mg of melatonin in comparison to control group [p< 0.001]. The results of this study demonstrate that, enriching the culture medium with melatonin, improve mouse preimplantation development

[Effect of fibroblastic growth factor on resumption of meiosis, in vitro maturation and embryo development of immature mouse oocytes]

The purpose of this study was to evaluate the effect of fibroblastic growth factor on resumption of meiosis, in vitro maturation of immature mouse oocytes and resulting embryo development with and without basic fibroblastic growth factor-4 [bFGF-4].Cumulus - oocyte complex [COCs] and germinal vesicle [GV] were obtained from female NMRI mice 46-48 hours after administration of an intra-peritoneal injection of 5 IU PMSG. COCs were cultured in TCM199 supplemented with different dosages of bFGF-4. After 24 hours, metaphase II [MII] oocytes were co-incubated with sperms for 4-6 hours in 16 medium. For all groups, the rate of cleaved embryos was assessed in the T6 medium until blastocyst stage. In all compared groups, the percentage of matured MII oocytes in the 10 ng/ml [%94.4] and 20 ng/ml [%92.5] of bFGF-4 treatment groups, was significantly higher [P<0.05] than those of the control group but the percentage of embryos that developed to blastocyst in 20 ng/ml bFGF-4 treatment group was significantly higher than those of the control group [P<0.05]. Exogenous bFGF-4 improved the oocyte maturation and embryo development