Conjugated linoleic acid stimulates apoptosis in RH and tehran strains of Toxoplasma gondii, in vitro

Background: The aim of this study was to evaluate the effects of conjugated linoleic acid (CLA) on apoptosis of tachyzoites of T. gondii, RH strain (type I) and the cyst-forming Tehran strain (type II) in vitro. Methods: Toxoplasma strains were injected into the peritoneal cavity of BALB/c mice. The Tehran strain forms cysts in the brain of mice. Bradyzoites within the cysts are reactivated to proliferative tachyzoites, by dexamethasone. Tachyzoites were aspirated from the peritoneum of infected mice, and the percentage of viable parasites was estimated with trypan blue staining. Tachyzoites were inoculated into HeLa cells cultivated in DMEM medium. Different concentrations of CLA were evaluated on T. gondii in HeLa cells by the tetrazolium (MTT) colorimetric assay. Differentiation between apoptosis and cell death was determined by flow cytometry using Annexin V and propidium iodide (PI) double staining. The statistical analysis performed by GraphPad Prism version 6.00. Results: CLA induces apoptosis in virulent (RH) and avirulent (Tehran) strains of T. gondii. The results of MTT indicated that CLA could decrease the proliferation of tachyzoites of both strains in HeLa cells. Conclusion: Conjugated linoleic acid has anti-toxoplasmacidal activity on tachyzoites of T. gondii. Therefore, we recommended further studies on this component in order to achieve a new drug against the parasite. © 2015, Tehran University of Medical Sciences (TUMS). All rights reserved

High resolution melting curve assay for detecting rs12979860 IL28B polymorphisms involved in response of iranian patients to chronic hepatitis C treatment

Background: A recent genome-wide association study (GWAS) on patients with chronic hepatitis C (CHC) treated with peginterferon and ribavirin (pegIFN-α/RBV) identified a single nucleotide polymorphism (SNP) on chromosome 19 (rs12979860) which was strongly associated with a sustained virological response (SVR). The aim of this study was twofold: to study the relationship between IL28B rs12979860 and sustained virological response (SVR) to pegIFN-α/RVB therapy among CHC patients and to detect the rs12979860 polymorphism by high resolution melting curve (HRM) assay as a simple, fast, sensitive, and inexpensive method. Materials and Methods: The study examined outcomes in 100 patients with chronic hepatitis C in 2 provinces of Iran from December 2011 to June 2013. Two methods were applied to detect IL28B polymorphisms: PCR-sequencing as a gold standard method and HRM as a simple, fast, sensitive, and inexpensive method. Results: The frequencies of IL28B rs12979860 CC, CT, and TT alleles in chronic hepatitis C genotype 1a patients were 10 (10/100), 35 (35/100), and 6 (6/100) and in genotype 3a were 13 (13/100), 31 (31/100), and 5 (5/100), respectively. In genotype 3a infected patients, rs12979860 (CC and CT alleles) and in genotype 1a infected patients (CC allele) were significantly associated with a sustained virological response (SVR). The SVR rates for CC, CT and TT (IL28B rs12979860) were 18, 34 and 4, respectively. Multiple logistic regression analysis identified two independent factors that were significantly associated with SVR: IL-28B genotype (rs 12979860 CC vs TT and CT; odds ratio ORs, 7.86 and 4.084, respectively), and HCV subtype 1a (OR, 7.46). In the present study, an association between SVR rates and IL28B polymorphisms was observed. Conclusions: The HRM assay described herein is rapid, inexpensive, sensitive and accurate for detecting rs12979860 alleles in CHC patients. This method can be readily adopted by any molecular diagnostic laboratory with HRM capability and will be clinically beneficial in predicting treatment response in HCV genotype 1 and 3 infected patients. In addition, it was demonstrated that CC and CT alleles in HCV-3a and the CC allele in HCV-1a were significantly associated with response to pegIFN-α/RBV treatment. The present results may help identify subjects for whom the therapy might be successful

Molecular epidemiology of epstein-barr virus (ebv) in patients with hematologic malignancies

Background: Epstein-Barr virus (EBV) is associated with different malignant diseases, such as Hodgkin lymphoma (HL) and lymphoproliferative disorders. Patients with hematologic malignancies by variable severity could be suspected for the infection with different types of this virus. This preliminary study reported the genotyping and related viral load of Epstein-Barr virus in Iranian patients with hematologic malignancies for estimation of possible factors affecting malignancy. Methods: Peripheral blood mononuclear cells (PBMC) of HL (n=20), NHL (n=29), acute lymphocytic leukemia (ALL) (n=18) and chronic lymphocytic leukemia (CLL) (n=12) were obtained. After DNA extraction, a nested-PCR and a conventional-PCR targeting EBNA-2 and EBNA-3C genes were performed. A real-time PCR assay for viral load quantitation carried out. Standard curve analysis used for evaluation of amplification specificity. Results: Of 79 included patients, 34 (43) were EBV positive. There were 23.5 (8/34), 38.2 (13/34), 23.5 (8/34), 14.8 (5/34) in HL, NHL, ALL and CLL groups, respectively. Also, the main genotype was genotype I (91.2) which it follows by 8.8 (3/34) genotype II. The real-time PCR assay showed the mean viral load ± std. deviation was 2.75�105 ± 1.202�106 copies/μg DNA and the higher viral load was seen in NHL patients. Conclusion: This preliminary investigation in Iran shows that the main EBV genotype into our region probably is genotype I (91.2) which it is similar to others. We could not find any statistically significant association between the virus infection and viral load with any specific disease and patients' demographic data. © 2020, Asian Pacific Organization for Cancer Prevention

Human Papillomavirus Type 16 Integration Analysis by Real-time PCR Assay in Associated Cancers

Human papillomavirus (HPV) is a common viral infection worldwide associated with a variety of cancers. The integration of the HPV genome in these patients causes chromosomal instability and triggers carcinogenesis. The aim of this study was to investigate the HPV-16 genome physical status in four major cancers related to HPV infection. Formalin-fixed paraffin-embedded blocks from our previous projects on head and neck, colorectal, penile, and cervical cancers were collected, and HPV-16�positive specimens were used for further analysis. The DNA extraction copy number of E2 and E7 genes was calculated by qualitative real-time PCR method. Serially diluted standards that were cloned in PUC57 plasmid were used. Standard curve and melting curve analysis was used for quantification. Of the 672 specimens studied, 76 (11.3) were HPV-16 positive. We found that 35.6 (16/45) were integrated. Statistical analysis showed that there were significant correlations between integration of HPV-16 and cervical cancer end-stage carcinogenesis (P <.0001), episomal form, and ASCUS lesions (P =.045). Significant correlation in penile cancer patients was seen between the episomal form and high-grade cancer stage (P =.037). Integration is a major factor in the carcinogenesis mechanism of HPV and has different prevalence in various cancers with a higher rate in progression except in penile cancer. © 201

Human Papillomavirus Type 16 Integration Analysis by Real-time PCR Assay in Associated Cancers

Human papillomavirus (HPV) is a common viral infection worldwide associated with a variety of cancers. The integration of the HPV genome in these patients causes chromosomal instability and triggers carcinogenesis. The aim of this study was to investigate the HPV-16 genome physical status in four major cancers related to HPV infection. Formalin-fixed paraffin-embedded blocks from our previous projects on head and neck, colorectal, penile, and cervical cancers were collected, and HPV-16�positive specimens were used for further analysis. The DNA extraction copy number of E2 and E7 genes was calculated by qualitative real-time PCR method. Serially diluted standards that were cloned in PUC57 plasmid were used. Standard curve and melting curve analysis was used for quantification. Of the 672 specimens studied, 76 (11.3) were HPV-16 positive. We found that 35.6 (16/45) were integrated. Statistical analysis showed that there were significant correlations between integration of HPV-16 and cervical cancer end-stage carcinogenesis (P <.0001), episomal form, and ASCUS lesions (P =.045). Significant correlation in penile cancer patients was seen between the episomal form and high-grade cancer stage (P =.037). Integration is a major factor in the carcinogenesis mechanism of HPV and has different prevalence in various cancers with a higher rate in progression except in penile cancer. © 201

Effects of morphine on replication of herpes simplex virus type 1 and 2

Several drugs are being used in treatment of HSV (Herpesviridae) infection in human but still introducing an effective safe drug is desirable. We investigated the inhibitory effect of morphine on replication of HSV in vitro. The results indicated that a concentration of up to 200 pg/ml morphine had a limited effect on Vero cell viability. At this concentration, the growth of HSV was inhibited considerably and after the third passage in presence of morphine it was completely eliminated. The presence of viral antigens in infected cells in presence of morphine by immunoflourescent staining showed that after the first passage a small number of infected cells contained viral proteins and at the third passage no cells with viral antigen was observed. This was confirmed by page and immunobloting techniques. Electron microscopy observation in cellular section indicated that there was no virus present in treated cells as compared with control untreated infected cells. © 2009 Academic Journals

The study of antiviral effects of Glycyrrihza Glabra extract on HSV

Backround: Recently, resistance to anti viral drugs has been reported. Hence, study of other components for obtaining new treatment approaches is necessary. Objective: The main objective of this study is to determine the inhibitory effect of Glycyrrihza Glabra on HSV replication. Methods: The first step was to evaluate the concentration of Glycyrrihza Glabra which was non toxic for Vero cells. Then, antiviral effects of Glycyrrihza Glabra in non toxic concentration zone were determined through TCID50 Method. IF method was also used in order to determine the reduction of viral proteins. Results: The results indicated that inhibitory effects of Glycyrrihza Glabra on replication of HSV are related to primary time of replication cycle. Conclusion: Glycyrrihza Glabra extract can be a suitable choice for treatment due to HSV infection

Serostatus of Epstein�Barr virus in Iranian MS patients

Multiple sclerosis (MS) is a common, complex disorder. Associations of MS with Epstein�Barr virus (EBV) infection in Caucasian populations are well documented. However, in the Iranian population, previous studies have been conflicting. Sixty patients with MS and 50 healthy controls were tested for anti-EBV antibodies using chemiluminescent assays. All MS patients to be seropositive for EBV as compared to 82 of controls (p = 0.0006). A strong, significant association of MS with EBV infection was documented, similar to studies in first world populations. Future studies should investigate the temporal sequence of infection to try to understand the cause of the recent increase in MS risk in Iran. © 2015, Belgian Neurological Society

Varicella zoster virus genotyping in chickenpox patient's clinical isolates from Iran

Aim: The varicella zoster virus (VZV) causes chickenpox and zoster infections. This study aimed to investigate the distribution of VZV genotypes among Iranian patients. Materials &amp; methods: From 2010 to 2015, 244 patients were enrolled in this cross-sectional study, 45 of whom were positive for VZV DNA. Both direct sequencing and restriction fragment length polymorphism assay were performed for 19 positive specimens. SPSS v.20 was used for statistics. Results: The predominant VZV genotype was M1 (84.2) followed by genotype E (10.5) and genotype J (5.3). Restriction fragment length polymorphism demonstrated that 17 strains were PstI+ BglI+ (M1 and/or J genotypes) and 2 were PstI+ BglI- (E genotype). Conclusion: This research is a prelim study on VZV genotyping. Further investigations will help to confirm the VZV genotype prevalence reported here. � 2016 Future Medicine Ltd