Inhibitory effect of Lactobacillus rhamnosus on pathogenic bacteria isolated from women with bacterial vaginosis

Background: Considering the high prevalence of bacterial vaginosis and its association with urinary tract infection in women and treatment of gynecologic problems occur when a high recurrence of bacterial vaginosis is often treated with antibiotics. The purpose of this study is to investigate the inhibitory effect of Lactobacillus rhamnosus on pathogenic bacteria isolated from women with bacterial vaginosis, respectively.Materials and Methods: 96 samples from women with bacterial vaginosis discharge referred to health centers dependent Shahid Beheshti University in 91-92 were taken by a gynecologist with a dacron swab and put in sterile tubes containing TSB broth and Thioglycollate broth and were immediately sent to the lab location in cold chain for the next stages of investigation. From Thioglycollate and TSB medium was cultured on blood agar and EMB and Palkam and Differential diagnosis environments, and then incubated for 24 h at 37°C. Strains of Lactobacillus rhamnosus were cultured in MRSA environment and were transfered to the lab. After purification of pathogenic bacteria, MIC methods and antibiogram, Lactobacillus rhamnosus inhibitory effect on pathogenic bacteria is checked. Statistical analysis was done by SPSS software v.16.Results: The results of this study show the inhibitory effect of Lactobacillus rhamnosus on some pathogenic bacteria that cause bacterial vaginosis, including Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus agalactiae, Entrococcus, Listeria monocytogenes and E.Coli. Microscopic examination of stained smears of the large number of Lactobacillus and pathogenic bacteria showed reduced. The prevalence of abnormal vaginal discharge, history of drug use means of preventing pregnancy and douching, respectively, 61%, 55%, 42% and 13% respectively. Significant difference was observed between the use and non-use of IUD in women with bacterial vaginosis infection.Conclusion: Our findings indicated the Inhibitory effect of Lactobacillus rhamnosus on the pathogenic bacteria that cause bacterial vaginosis. The results of this study confirm the hypothesis of inhibit of pathogens growth that cause bacterial vaginosis supported by probiotics and can have beneficial effects in the prevention and treatment of bacterial vaginosis

Genotyping of Virulence Factors of Uropathogenic Escherichia coli by PCR

Background: Escherichia coli is the most causative agent of urinary tract infections (UTIs). Apart from all human infectious diseases, UTI have a high prevalence and in most cases, Escherichia coli is a dominance bacterium which can cause pyelonephritis and cystitis. The aim of the study was to determine the occurrence of some virulence genes expressing fimbriae, production of hemolysin and aerobactin among a hundred Escherichia coli isolates obtained from in-and outpatients of Karaj Shahid Rajaii hospital, showing clinical and laboratory signs of UTI.Materials and Methods: In this investigation we isolated Escherichia coli strains from urine samples of patients with UTI during the period of July to December 2012 and studied them for the presence of the virulence genes by PCR.Results and Conclusion: The most abundant virulence factor in this study was fimH. The prevalence of the virulence factors for fimbriae type 1 (fimH gene), pyelonephritis associated pili (pap gene), S-family adhesions (sfa gene), hemolysin (hly gene) and aerobactin (aer gene), was 73%, 46%, 32%, 47%, 57%, respectively

The Inhibitory Effects of Lactobacillus Reuteri’s Cell Wall on Cell Proliferation in the HCT-116 Colorectal Cancer Cell Line

Background: Colorectal cancer (CRC) is the third and a second common cancer in men and women respectively in the world and about 1.4 million new cases diagnosed in 2012. The normal gut microflora consists of bacterial species. One group of them is probiotics, which confer a health benefit to the host. Lactobacillus reuteri (L.reuteri) is known as a probiotic, which lead to the prevention of colorectal cancer. The aim of this study was to assess the inhibitory effects of Lactobacillus reuteri’s cell wall on cell proliferation in the colorectal cancer HCT-116 cell line.Materials and Methods: The cells of HCT-116 cell line were grown at 37ᵒC, 5% CO2. L.reuteri was obtained from the Iranian Biological Resource Center and cultured in the MRS Broth at 37ᵒC for 48h anaerobically. The cell wall was prepared by the freezing-thawing procedure. So the inhibitory effect of L.reuteri on the growth and proliferation of HCT-116 cells was assessed by MTT assay.Results: The cell wall from L.reuteri inhibited cell proliferation on colorectal cancer HCT-116 cell line. It showed dose- and –time dependent inhibition.Conclusion: These results demonstrated that cell wall of L.reuteri inhibits cell proliferation of HCT-116 cell line

Antimicrobial Activity of Three Root Canal Irrigants on Enterococcus Faecalis: An in Vitro Study

INTRODUCTION: The aim of this study was to compare the antimicrobial effects of 2.5% Sodium hypochlorite (NaOCl), 2% Chlorhexidine Gluconate (CHX) and BioPure MTAD (MTAD) on Enterococcus (E) faecalis-contaminated root canals of human extracted teeth. MATERIALS AND METHODS: Seventy human intact extracted single-rooted teeth with straight root canal randomly divided into 5 groups: positive control (n=5), negative control (n=5), 2.5% NaOCl (n=20), 2% CHX (n=20), and MTAD (n=20). Each tooth was instrumented using the passive step-back technique hand and rotary instruments. E. faecalis incubated into the canals and grew for 4 weeks. Canals irrigated using three mentioned solutions for 5 minutes. Samples were taken from canal walls and transferred into Brain Heart Infusion (BHI) culture medium and placed in an incubator at 37ºC for 96 hours and bacteriological evaluations were done. Chi- Square test and SPSS software were used for the statistical analysis of the results. RESULTS: Bacterial growth was seen in only one sample of MTAD group (5%), but in 4 of CHX group (20%) and 5 of NaOCl group (25%). Chi-Square test showed no statistically difference between groups. CONCLUSION: Based on the results of this study, it seems that all three solutions have acceptable antimicrobial effect on E. faecalis

Detection of actA and InlB genes in Listeria monocytogenes Isolated from women with Spontaneous abortions

 AbstractBackground & Objective To assess the extent of L. monocytogenes in causation of human spontaneous abortions by isolation methods and PCR analysis for the presence of actA and InlB genes.Methods : In this descriptive study, vaginal swabs were collected from 96 women with spontaneous abortions who referred to Tehran`s hospitals. The samples were cultured in to Listeria specific media (PALCAM agar). Then , the Listeria genus was verified by differentiation  biochemical tests, such as, haemolysis on Blood agar, Catalase and Oxidase reaction, motility at room temperature. PCR test was performed on all samples and detected the actA and InlB genes of L. monocytogenes.Results : In culture, 7 of 96 samples were positive for L.monocytogenes.                                     in PCR technique, actA and InlB genes were detected from 12 and 2 vaginal samples,  respectively.Conclusions : The occurrence of pathogenic L. monocytogenes in cases of spontaneous abortions was 12.5%. It seems that the actA and InlB genes have an important roles as essential virulences determinant in pathogenic L.monocytogenes. The results show the PCR method is more sensitive, easier and faster than culture to detect L.monocytogenes.Key word : Listeria monocytogenes , spontaneous abortions , actA gene , InlB gen

Cytoplasmic and membranous CD24 marker expression has indirect correlation with cAMP/cGMP ratio

Background- CD24 is a cell adhesion molecule that has been implicated in metastatic tumor progression cells. Our aim was clarify correlation between CD24 expression and cAMP:cGMP ratio in murine colorectal cancer cell line (CT26) after using cholera toxin.Materials and Methods- The CT26 cells were cultured in microtubes for assaying cAMP and cGMP; also the cells were cultured in flasks for assaying cytoplasmic and membranous CD24 expression. The Real-Time PCR was done for cDNA that was synthesized from CT26 cells’ mRNA. Also, expression CD24 marker of cells was determined by Anti-CD24 antibody and Goat Anti-Rabbit IgG-FITC (flow cytometry).Results- The cholera toxin grew cAMP:cGMP ratio and it influenced cytoplasmic and membrane CD24 expression.Conclusion- There are indirect correlation between cAMP:cGMP ratio and CD24 expression

Evaluation of Antimicrobial Activity of Cuminum Cyminum Essential Oil and Extract against Bacterial Strains Isolated from Patients with Symptomatic Urinary Tract Infection

Background: Many efforts have been done to find effective agents against resistant pathogens. Cuminum cyminum L. (Cumin) is an aromatic plant within the Apiaceae family. It has a variety of purposes and demonstrates antimicrobial and antioxidant properties. This study evaluated the activity of C. cyminum extract and essential oil against bacterial isolates which cause urinary tract infection, including Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus agalactiae, group A streptococci, Enterococcus faecalis, Staphylococcus epidermidis, Staphylococcus aureus and Staphylococcus saprophyticus isolated from patients with urinary tract infection.Materials and Methods: Extract was prepared by maceration and essential oil was prepared by hydrodistillation from C. cyminum seeds. The study population was 95 patients with urinary tract infection without malignant diseases, diabetes and immunosupression. After identification of organism, susceptibility testing was carried out by disc diffusion method and MIC values by broth microdilution testing.Results: C. cyminum essential oil can have a better effect on the gram-negative bacteria causing urinary tract infection than gram-positive bacteria. In addition, C. cyminum extract have good activity against both gram- positive and gram-negative bacteria. Our findings also showed that essential oil and extract of C. cyminum has better antibacterial activity on uropathogen isolates than amoxicillin and the difference was significant (P value<0.05) but the activity is not superior to other antibiotics.Conclusion: These results suggest that the essential oil and extract of C. cyminum seeds might be considered as interesting sources of antibacterial components against uropathogenic bacteria

Detection of acrA, acrB, aac(6′)-Ib-cr, and qepA genes among clinical isolates of Escherichia coli and Klebsiella pneumoniae

Background: The distribution of drug resistance among clinical isolates of Escherichia coli and Klebsiella pneumoniae has limited the therapeutic options. The aim of this study was to report the prevalence of quinolone resistance genes among E. coli and K. pneumoniae clinical strains isolated from three educational hospitals of Tehran, Iran. Materials and methods: A total of 100 strains of E. coli from Labbafinejad and Taleghani Hospitals and 100 strains of K. pneumoniae from Mofid Children and Taleghani Hospitals were collected between January 2013 and May 2014. Antimicrobial susceptibility tests were done by disk diffusion method based on Clinical and Laboratory Standards Institute guidelines. Detection of qepA, aac(6′)-Ib-cr, acrA, and acrB genes was done by polymerase chain reaction (PCR). Results: In this study, fosfomycin and imipenem against E. coli and fosfomycin and tigecycline against K. pneumoniae had the best effect in antimicrobial susceptibility tests. PCR assay using specific primers demonstrated that the prevalence of qepA, aac(6′)-Ib-cr, acrA, and acrB genes among the 100 E. coli isolates was 0 (0%), 87 (87%), 92 (92%), and 84 (84%), respectively. The prevalence of qepA, aac(6′)-Ib-cr, acrA, and acrB genes among the 100 K. pneumoniae isolates was 4 (4%), 85 (85%), 94 (94%), and 87 (87%), respectively. Conclusion: The distribution of qepA, aac(6′)-Ib-cr, acrA, and acrB resistance determinants in E. coli and K. pneumoniae is a great concern. Therefore, infection control and prevention of spread of drug-resistant bacteria need careful management of medication and identification of resistant isolates

RAPD PCR Profile, Antibiotic Resistance, Prevalence of armA

The increasing prevalence of multidrug-resistant Klebsiella pneumoniae strains isolated from hospitals shows the limitation of recent antibiotics used for bacterial eradication. In this study, 81 K. pneumoniae isolates were collected from three hospitals in Tehran. Antibiotic susceptibility test showed the highest rates of resistance to cefotaxim (85.5%) and ceftazidime (78.3%), and the lowest rates of resistance were detected for colistin (16.9%), streptomycin (16.8%), and chloroamphenicol (21.7%). Eleven different resistance patterns were observed. Sixty-six out of 81 isolates (81.5%) were found to be multidrug resistant (MDR), and 35.8% of them belonged to A3 resistance pattern. 7.4% and 66.7% were KPC enzyme and armA gene positive, respectively. RAPD PCR assay of these bacteria showed 5 clusters, 16 single types, and 14 common types, and there was not any correlation between genetic patterns of the isolates and presence of resistance agents. Simultaneous detection of resistance-creating agents could be an important challenge for combination therapy of MDR K. pneumoniae-caused infections

Prevalence of Chlamydia trachomatis and Mycoplasma genitalium in Patients with Benign and Malignant Ovarian Cancer by Nested PCR Method

Background: Chlamydia trachomatis (C. trachomatis) and Mycoplasma genitalium (M. genitalium) are considered factors in cervical and ovarian cancer and are associated with flaky cell carcinoma of the cervix. The role of steady infection, leading to chronic inflammation, in the of ovarian cancer has received very little consideration, although a background of pelvic inflammatory disease (PID) is in a case-control study associate to higher risk for ovarian cancer. C. trachomatis, the most common and important cause of PID in the developed world is the genital and cervical infectious agent. The aim of this study was prevalence of C. trachomatis and M. genitalium in patients with ovarian cancer who referred to Imam Hossein Hospital of Shahid Beheshti University of Medical Sciences, Tehran, Iran.Materials and Methods: In this descriptive study that was conducted from January 2014 to April 2015, 124 samples were studied which obtained from patients with ovarian cancer who referred to medical centers of Shahid Beheshti University of Medical Sciences. After obtaining samples from ovarian cancer tissue by the pathologist, for extraction DNA, samples were transferred to the laboratory of university. To confirm the presence of C. trachomatis in samples of ovarian cancer, specific primers for the Major Outer Membrane Protein (MOMP) genes of C. trachomais, were designed and used Nested PCR method for detection of M. genitalium. Sequencing was performed on the PCR and Nested PCR product to confirm the presence of C. trachomatis and M. genitalium.Results: Out of 124 samples of ovarian cancer, 62 (50%) samples were malignant cancer and 62 (50%) were benign cancer as control group. From 65 malignant samples 14 (22.5%) were Chlamydia trachomatis positive. None of the tissue samples of benign cancer of ovary were positive for C. trachomatis. Notably, none of the 124 ovarian samples were positive in the M. genitalium standard PCR assay.Conclusion: The results suggest that the spread of C. trachomatis in the female with ovarian cancer may be common. This finding reflects a possible role of C. trachomatis in the carcinogenesis of ovarian tumors. C. trachomatis infection may play a relative role in the pathogenesis of ovarian carcinomas or it could facilitate its progression