Antiviral effects of Lactobacillus crispatus against HSV-2 in mammalian cell lines

Background: Herpes simplex virus type 2 (HSV-2) infectious disease is one of the most common viral sexually transmitted diseases. As regards, vaginal lactobacilli play an important role in protecting host against the urogenital pathogens; here we assessed the potential antiviral activity of Lactobacillus crispatus against HSV-2 infection in vitro. Methods: Both Vero and HeLa cell lines were treated by L. crispatus before, during and after HSV-2 infection. The pre-incubation assay was also performed for the evaluating of virus adsorption by L. crispatus. Virus titer reduction in each stage was determined by a plaque reduction assay. Results: L. crispatus significantly decreased the infectivity of the HSV-2 in initial steps on both cell lines; however, no significant inhibition was ascertained during adsorption and multiplication process. The lactobacilli adhere on Vero cells two-fold stronger than HeLa and subsequently protect the Vero cells nearly 2.5 fold higher than HeLa cell against the virion. Co-incubation of HSV-2 with bacterial cells prior to virus inoculation significantly decreased the virus titer. Conclusion: L. crispatus appears to inhibit the entry of the virus into cells by trapping HSV-2 particles. In addition, formation of L. crispatus microcolonies in the cell surface could block HSV-2 receptors and prevent viral entry to cells in initial infection steps. © 201

Prevalence of hepatitis G virus among hemodialysis and kidney transplant patients in Khuzestan Province, Iran

Background: Hepatitis G virus (HGV) is a member of Flaviviridae. Prevalence of HGV in healthy people is very low, but this virus is more prevalent in patients with hepatitis. Besides, relative frequency of HGV in patients undergoing hemodialysis, and kidney recipients is very high. The role of HGV in pathogenesis is not clear. Since this virus cannot be cultivated, molecular techniques such as Revers Transcription Polymerase Chain Reaction (RT-PCR) is applied to detect HGV. Objectives: The current study aimed to investigate the prevalence of HGV using determination of E2, viral envelope antigen, antibodies and the RNA by Enzyme Linked Immunosorbent Assay (ELISA) and RT-PCR techniques. The rational of the study was to determine the prevalence of HGV in patients undergoing hemodialysis and kidney transplantation in Khuzestan province, Iran. Patients and Methods: Five hundred and sixteen serum samples of the patients undergoing hemodialysis and kidney transplantation from various cities of Khuzestan province were collected. Anti-hepatitis G E2 antibodies were investigated by ELISA method. RNAs were extracted from serums and Hepatitis G RNA was detected by RT-PCR. Results: Of the 516 samples, 38 (7.36) specimens were positive for anti-HGV by ELISA. All of these ELISA positive samples were negative for HGV genome by RT-PCR. Of the remaining 478 ELISA negative samples, 16 (3.14) samples were positive by RT-PCR. Conclusions: Hepatitis G Virus was not prevalent in the patients undergoing hemodialysis and kidney transplantation in Khuzestan province. Although reports indicated high frequency of co-infection of HGV with hepatitis B and C viruses, in the current research, co-infection of HGV with B and C was not considerable. Since diferent groups and subtypes of HGV are reported, periodic epidemiologic evaluation of HGV and its co-infection with other hepatitis viruses is suggested in other populations such as the patients with thalassemia; however, periodic epidemiologic monitoring of HGV may be helpful to control future potential variations of the virus. © 2015, Ahvaz Jundishapur University of Medical Sciences

In vitro adherence of Lactobacillus strains isolated from the vaginas of healthy Iranian women

Background The lactobacilli are a part of the bacterial flora of the human vagina. Detection of normal Lactobacillus species in the vaginas of healthy women in different geographical locations, and evaluation of their specific properties, can aid in the selection of the best species for preventing sexually transmitted diseases in the future. This study was performed to isolate and identify the Lactobacillus species in the vaginas of healthy women and to evaluate the adherence of these lactobacilli to Vero and HeLa cell lines. Methods The study included 100 women. Bacteria were isolated from healthy women and purified. Phenotypic and biochemical tests were performed to identify the lactobacilli. The Lactobacillus species were detected by molecular methods using polymerase chain reaction amplification of the full length of the 16S rDNA of the isolated bacteria. Several isolates of each species were then selected to study their adherence to Vero and HeLa cell lines. Results Among the 50 samples taken from healthy women meeting the inclusion criteria, Lactobacillus species were identified in 33 (66) samples. Of these lactobacilli, 14 isolates were Lactobacillus crispatus, six (18.2) were Lactobacillus gasseri, nine (27) were Lactobacillus rhamnosus, and the rest were either Lactobacillus salivarius (6) or Lactobacillus plantarum (6). L. rhamnosus showed the greatest adhesion to the cells when compared to the other tested species. All the lactobacilli isolated in this study showed a smaller capacity for cell adherence when compared with control species. Conclusion L. crispatus, L. rhamnosus, and L. gasseri were the dominant Lactobacillus species in the vaginas of healthy women in Iran. L. rhamnosus attached more readily to the cells than did the other species; therefore, this isolate is a good candidate for further studies on the potential health benefits and application of lactobacilli as probiotics. © 201

High prevalence of dna from non-H. pylori helicobacters in the gastric mucosa of venezuelan pet dogs and its histological alterations

Non-H. pylori helicobacters (NHPH) have been demonstrated as gastric spiral-shaped bacteria in specimens obtained from dogs; however, their roles in the pathogenesis of upper gastrointestinal disease have not yet been clearly established. The purpose of this study was to evaluate the prevalence of NHPH DNA in the gastric mucosa of dogs and its association with histopathology. Helicobacter was detected through histopathological techniques, PCR, and FISH analysis from fundic biopsies of twenty dogs with or without signs of gastrointestinal disease. PCR and FISH were based on partial 16S rRNA gene sequences. Nineteen dogs showed mild to marked gastritis in the fundus, and only one dog had a healthy gastric mucosa. NHPH DNA was detected in 18 dogs with gastritis and one with normal gastric mucosa. However, there was no significant correlation between the presence of NHPH DNA and the degree of gastritis. These results show a high prevalence of NHPH DNA in the gastric mucosa of dogs from Venezuela. Further studies are necessary to determine a possible association between a specific NHPH species and the degree of gastritis

Identification of Cutaneous Leishmaniasis Agents by Nested Po-lymerase Chain Reaction (Nested-PCR) in Shush City, Khuzestan Province, Iran

Background: Leishmaniasis is a common parasitic disease world wide. Leishmania tropica and L. major are two common cause of cutaneous leishmaniasis in Iran. The aim of this study was determination of the cause of cutaneous leishmaniasis in Shush city, Khouzestan Province, Southwest Iran"nMethods: One hundred samples were collected from patients at the age of 1-80 year with documented cutaneous leishmani¬asis referred to the health centre and a private medical diagnostic laboratory at Shush City. DNA was extracted from slid samples by phenol- chloroform- Isoemil alcohol method, and subjected to Nested-PCR as template. k DNA of the parasites were amplified by CSB1XR and CSB2XF in the first round of PCR and 13Z and Li R primers for the second round. After PCR, electrophoresis of products was performed and 750bp band from L. tropica and 560bp band from L. major were de¬tected."nResults: A total of 100 cases comprising 47 females and 53 males were studied. The highest infected age group was under 10 years with a rate of 42% and the lowest rate was 4% at the age group of above 40 years. The results of PCR electropho-resis indicated that 90(90%) cases were L. major and 10 (10%) L. tropica. The predominant species in this area was L. ma-jor."nConclusion: It is concluded that Nested PCR is a reliable test for diagnosis and identification of Leishmania species and can apply in epidemiological investigations

Bioconjugated fluorescent silica nanoparticles for the rapid detection of Entamoeba histolytica

Rapid detection of Entamoeba histolytica based on fluorescent silica nanoparticle (FSNP) indirect immunofluorescence microscopy was evaluated. Silica nanoparticles were synthesized using Stöber's method, with their surface activated to covalently bind to, and immobilize, protein A. For biolabeling, FSNP was added to conjugated E. histolytica trophozoites with monoclonal anti- E. histolytica IgG1 for microscopic observation of fluorescence. Fluorescent silica nanoparticle sensitivity was determined with axenically cultured E. histolytica serially diluted to seven concentrations. Specificity was evaluated using other intestinal protozoa. Fluorescent silica nanoparticles detected E. histolytica at the lowest tested concentration with no cross-reaction with Entamoeba dispar, Entamoeba moshkovskii, Blastocystis sp., or Giardia lamblia. Visualization of E. histolytica trophozoites with anti- E. histolytica antibody labeled with fluorescein isothiocyanate (FITC) was compared with that using anti- E. histolytica antibody bioconjugated FSNP. Although FITC and FSNP produced similar results, the amount of specific antibody required for FITC to induce fluorescence of similar intensity was fivefold that for FSNP. Fluorescent silica nanoparticles delivered a rapid, simple, cost-effective, and highly sensitive and specific method of detecting E. histolytica. Further study is needed before introducing FSNP for laboratory diagnosis of amoebiasis. © 2015 Elsevier B.V

Riboflavin may ameliorate neurological motor disability but not spatial learning and memory impairments in murine model of multiple sclerosis

Summary: Background & aims: Riboflavin has an important role in myelin formation. This experimental study assesses the interactions between the effects of riboflavin and interferon beta-1a (INF-β1a) on motor disability, spatial learning and memory, and brain-derived neurotrophic factor (BDNF) in experimental autoimmune encephalomyelitis (EAE). Methods: In the present research C57BL/6 mice (n = 56) were divided into sham and treatment groups. Riboflavin was administrated (10 mg/kg/day) orally for two weeks alone and/or combined with INF-β1a at 150 IU/g of body weight. After the induction of EAE, the animals were investigated for the clinical signs. Spatial learning and memory were assessed through the standard Morris water maze (MWM). The brain and spinal cord levels of BDNF were studied using real-time polymerase chain reactions and enzyme-linked immunosorbent assay. The data were analyzed using one-way ANOVA, repeated measures, and generalized estimating equations model. Results: The results in the brain revealed that BDNF mRNA expression (P < 0.01) and protein levels (P < 0.05) increased in the EAE mice treated with the combination of riboflavin and INF-β1a compared to the treated groups with riboflavin or INF-β1a. Clinical scores were reduced in groups treated with riboflavin compared to other groups. EAE mice treated with riboflavin swam significantly faster in MWM compared to other groups (P < 0.05). No significant differences were found between EAE and healthy mice in other spatial learning and memory evaluating variables. Conclusion: The data highlighted the synergistic role of riboflavin and INF-β1a in improving the disability but not spatial learning and memory mediated by BDNF in EAE. Keywords: Riboflavin, Experimental autoimmune encephalomyelitis, Brain-derived neurotrophic factor, Memor