Characterization of Vaginal Lactobacilli with Potential Probiotic Properties Isolated from Healthy Women in Northern Iran.

Vaginal lactobacilli protect against bacterial vaginosis and vaginal candidiasis. They may have probiotic properties and help maintain the balance and health of the vaginal ecosystem while the loss of these bacteria predisposes females to urinary and genital infections. The aim of this study was to investigate the probiotic potential of vaginal Lactobacillus among healthy females in northern Iran. The Lactobacillus strains were isolated from vaginal samples and were identified by sequencing of the 16S rRNA fragment. Functional properties such as tolerance to low pH, H2O2 production, adherence ability to Hela cells and antagonistic activity against Candida albicans was examined. A total of 38 vaginal lactobacilli strains from five species, including Lactobacillus crispatus (n = 13), Lactobacillus gasseri (n = 10), Lactobacillus acidophilus (n = 6), Lactobacillus jensenii (n = 5) and Lactobacillus johnsonii (n = 4), were identified. All of the species showed significant tolerance to low pH over 24 h (p < 0.001). The best adherence ability to Hela cells was seen in Lactobacillus gasseri strains. Nearly 17 of the strains had higher anti-candida activity compared to the other strains. According to the findings, four lactobacilli strains isolated in the vaginal samples of healthy Iranian women had the best probiotic potential.

The phenotypic and genotypic antibiotic susceptibility of vaginal Lactobacillus with potential probiotic properties isolated from healthy women in northern Iran.

Background and Objectives: This study assesses the antibiotic susceptibility of vaginal Lactobacillus strains and provides data for determining the prevalence of certain antibiotic resistance genes in the new strains of lactobacilli serving as probiotics and selected from healthy women in northern Iran. Materials and Methods: One hundred premenopausal non-pregnant women in the reproductive age range of 22-50 years participated in this study. The potential probiotic vaginal lactobacilli used in the study included Lactobacillus crispatus (34.2%), Lactobacillus gasseri (26.3%), Lactobacillus johnsonii (10.5%), Lactobacillus acidophilus (15.7%) and Lactobacillus jensenii (13.1%). The phenotypic antibiotic susceptibility of the strains was determined by E test and DNA extraction and PCR were performed to examine the antibiotic resistance genes. Results: 38 potential probiotic vaginal lactobacilli were isolated. All the strains of lactobacilli were resistant to metronidazole and trimethoprim/sulfamethoxazole and all of the strains were susceptible to ampicillin and chloramphenicol antibiotics. The results showed that ermB, ermC, and ermA genes were observed in the strains of Lactobacillus acidophilus. Metronidazole resistance (nim) gene was also found in one strain of Lactobacillus crispatus and Lactobacillus johnsonii. The aminoglycoside resistance (aac6'-aph2″) gene was observed in 8% of the strains. Also, tetM, tetK and tetW genes were found in more than 80% of the Lactobacillus strains. Conclusion: The antimicrobial susceptibility of vaginal lactobacilli is an important criterion for establishing whether or not the organism is a probiotic. A high level of resistance to clinical antibiotics, such as metronidazole and aminoglycosides, was demonstrated. Antibiotic resistant genes also appeared widely in vaginal lactobacilli.

Evaluation of Anti-Tumor Potential of Lactobacillus acidophilus ATCC4356 Culture Supernatants in MCF-7 Breast Cancer.

BACKGROUND: The anti-cancer activity of some lactic acid bacterial strains is well documented in several kinds of literatures. Lactobacillus strains have received considerable attention as a beneficial microbiota. The aim of this study is to evaluate the effects of anti-tumor activities of L. acidophilus ATCC4356 culture supernatants on the MCF-7 human breast cancer cells. MATERIALS AND METHODS: The anti-cancer effects of 24h and 48h culture supernatants at various concentrations (1.25, 2.5, 5, 10 and 20 µg/ml) were determined by various in vitro and in vivo assays including MTT, tumor volume measurement as well as 99mTc-MIBI biodistribution in MCF-7 tumor bearing nude mice and histopathology test. For evaluation of the related mechanism of action, quantitative PCR was conducted. RESULTS: The 48h culture supernatants at 10 and 20 µg/ml exhibited significant in vitro inhibition of MCF-7 cell proliferation. However, this inhibition was not observed for HUVEC human endothelial normal cells. Q-PCR indicated that treatment by the supernatant led to a significant downregulation of VEGFR (~ 0.009 fold) and Bcl- 2 (~ 0.5 fold) and upregulation of p53 (~ 1.3 fold). In vivo study using MCF-7 xenograft mouse models demonstrated a reduction in tumor weight and volume by both 24h and 48h supernatants (2 mg/kg) after 15 days. According to the 99mTc-MIBI biodistribution result, treatment of MCF-7 bearing nude mice with both 24h and 48h supernatant (2mg/kg) led to a significant decrease in tumor uptake compared with the control group. CONCLUSION: These results suggest that the culture supernatants of L. acidophilus ATCC4356 at suitable concentrations can be considered as a good alternative nutraceutical with promising therapeutic indexes for breast cancer.

Application of Epstein-Barr Virus for Optimization of Immortalized B-lymphocyte Production as a Positive Control in Genetic Studies.

BACKGROUND: Infection of B-cells with Epstein-Barr virus (EBV) leads to more and subsequent immortalization. This is considered as the method of choice for generating lymphoblastoid cell lines (LCLs). Producing LCLs, although very useful but is very time consuming and troublesome, drives the requirement for quicker and more reliable methods for EBV-driven B-cell transformation. MATERIALS AND METHODS: After successfully production of LCLs, different parameters including temperature, serum concentration, type of culture medium, and CO2 concentration were evaluated on EBV-transformed B-cells. In this study, we were able to produce LCLs and optimize condition. RESULTS: The best condition for generating LCLs was 37°C, 5% CO2, 20% fasting blood sugar, and RPMI 1640. The study results were to establish a reliable method for producing LCLs that can be used to produce immortalized B-cells from almost any sources. CONCLUSION: This can help with tumorgenecity studies, as well as producing control material for rare genetic disorders and so on. The aim of this study was to determine optimized condition for reliable and reproducible LCLs from different sources.

A Comparative Serological Study of Toxoplasmosis in Pregnant Women by CLIA and ELISA Methods in Chalus City Iran.

BACKGROUND: Toxoplasmosis is the most common disease in humans and animals (zoonosis) caused by the protozoan parasite Toxoplasma gondii. The disease is usually appeared as asymptomatic in immunocompromised individuals but its most common symptom is lymphadenopathy. Shortly before or during the first trimester of pregnancy, this disease can be transferred to the fetus and cause serious infection in the fetus. In late pregnancy (third trimester), the complications of this infection is very low or unsigned. Due to the absence of non-specific clinical symptoms or slight infection in pregnant women, prenatal diagnosis is often impossible. OBJECTIVES: Since no research compared these two methods, we decided to compare these methods and determine which method works better for diagnosis of toxoplasmosis. PATIENTS AND METHODS: In this study, 50 pregnant women who referred to the Chalus Health Center laboratory were included and the blood samples were tested for presence of IgG and IgM antibodies of Toxoplasma gondii by both ELISA and Chemiluminescence methods. RESULTS: Of the 50 samples tested by the ELISA method, 26 samples (52%) were positive for IgG . No samples were positive for IgM. Of the 50 samples tested by the Chemiluminescence method, 28 samples (56%) were positive for IgG. No samples were positive for IgM. CONCLUSIONS: A significant relationship between the age of the youngest child and the infection rate was seen. No significant correlation between age, number of individuals in the household, number of children, location, type of construction, consumption of greens, the way of greens and meat consumption, drug use, history of stillbirth and infection levels was seen.