Low viral load of Merkel cell polyomavirus in Iranian patients with head and neck squamous cell carcinoma: Is it clinically important?

Recent studies show that the human Merkel cell polyomavirus (MCPyV) may be involved in causing cancer. The objective of this study was to assess the impact of MCPyV on the development of head and neck squamous cell carcinoma (HNSCC). In total, 50 paraffin-embedded HNSCC biopsy samples and 50 adjacent non-cancerous samples were evaluated for the presence of MCPyV DNA and RNA. Among patients, the five most frequent histopathologic sites were the tongue (22.0%), lip (16.0%), submandibular (14.0%), cheek (14.0%), and throat (14.0%). MCPyV DNA was positive in eight (16.0%) samples. The median MCPyV LT-Ag copy number in the eight positive samples and in one non-cancerous sample was 4.8 × 10-3 and 2.6 × 10-5 copies/cell, respectively. Quantification of MCPyV LT-Ag revealed increased expression in stage III (5.6 × 10-3 copies/cell) than in the other stages. The MCPyV DNA load in different stages of HNSCC was also statistically significant (P = 0.027). The viral load was low, suggesting that only a fraction of cancerous cells is infected. This result provides evidence confirming the presence of MCPyV in a subset of Iranian patients with HNSCCs, but further studies needed to confirm our findings.

Comparison of Three Different Methods for Detection of IL28 rs12979860 Polymorphisms as a Predictor of Treatment Outcome in Patients with Hepatitis C Virus.

OBJECTIVES: This study aimed to evaluate the specificity, sensitivity, cost, and turn-around time of three methods of gene polymorphism analysis and to study the relationship between IL28B rs12979860 and SVR rate to pegIFN-α/RVB therapy among patients with chronic hepatitis C. METHODS: A total of 100 samples from chronic hepatitis C patients were analyzed in parallel using the three methods: direct sequencing, real-time polymerase chain reaction (PCR), amplification refractory mutation system (ARMS)-PCR. RESULTS: The different profiles for IL28B rs12979860 alleles (CC, CT, and TT) obtained with PCR-RFLP, ARMS-PCR, and direct sequencing were consistent among the three methods. Prevalence of rs12979860 genotypes CC, CT and TT in HCV genotype 1a was 10(19.6%), 35(68.6%), and six (11.8%), respectively, and in HCV genotype 31, it was 13(26.5%), 31(63.3%), and five (10.2%), respectively. No significant difference was seen between rs12979860 genotype and HCV genotype (p = 0.710). CONCLUSION: Screening by ARMS - PCR SNOP detection represents the most efficient and reliable method to determine HCV polymorphisms in routine clinical practice.

Novel effects of Helicobacter pylori CagA on key genes of gastric cancer signal transduction: a comparative transfection study.

Helicobacter pylori (H. pylori) infection is now recognized as a worldwide problem. Helicobacter pylori CagA is the first bacterial oncoprotein to be identified in relation to human cancer. Helicobacter pylori CagA is noted for structural diversity in its C-terminal region (contains EPIYA motifs), with which CagA interacts with numerous host cell proteins. Deregulation of host signaling by translocated bacterial proteins provides a new aspect of microbial-host cell interaction. The aim of this study is to compare the cellular effects of two different CagA EPIYA motifs on identified signaling pathways involve in gastric carcinogenesis. To investigate the effects of CagA protein carboxyl region variations on the transcription of genes involved in gastric epithelial carcinogenesis pathways, the eukaryotic vector carrying the cagA gene (ABC and ABCCC types) was transfected into gastric cancer cell line. The 42 identified key genes of signal transduction involved in gastric cancer were analyzed at the transcription level by real-time PCR. The results of real-time PCR provide us important clue that the ABCCC oncoprotein variant can change the fate of the cell completely different from ABC type. In fact, these result proposed that the ABCCC type can induce the intestinal metaplasia, IL-8, perturbation of Crk adaptor proteins, anti-apoptotic effect and carcinogenic effect more significantly than ABC type. These data support our hypothesis of a complex interaction of host cell and these two different H. pylori effector variants that determines host cellular fate.

Methanobrevibacter smithii archaeosomes-entrapped mzNL4-3 virus-like particles induce specific T helper 1-oriented cellular and humoral responses against HIV-1.

Despite numerous and tremendous achievements in the development and standardization of HIV vaccines, there are still lots of vague concepts in HIV vaccinology. Various approaches have been applied to design an efficient HIV vaccine. Due to their lack of replication ability and expression of native antigens at the same time virus-like particles, such as previously introduced mzNL4-3 HIV-1 VLPs are among the highlighted candidates in this field. On the other part, application of adjuvants is an inseparable strategy in the vaccine development researches. Archaeosomes are liposomal adjuvants with intensifiying features of T helper 1 and cytotoxic T-cells responses. Archaeosomes derived from Methanobrevibacter smithii has been shown to enhance MHC class I-dependent antigen presentation and hence, are to be advantageous in the development of vaccines against viral infections. Herein, we have studied efficiency of mzNL4-3 VLPs entrapped in M. smithii archaeosomes as an HIV-1 vaccine candidate to induce humoral and cellular responses in BALB/c mice. Analysis of total and subtype-specific anti-Env IgG antibody, as well as, cytokine secretion pattern revealed an efficient promotion of anti-HIV specific T helper 1 responses in immunized animals. This finding was evidenced by the significant dominance of IgG2a subtype in the sera and considerable secretion of IFN-γ by specifically induced splenocytes of mice immunized with VLP-containing archaeosomes (VLP+ Archaeosome). In addition, ELISpot assay verified these results and indicated the significantly higher frequency of IFN-γ secreting splenocytes in immunized models. The ratio of IFN-γ to IL-4 spot forming cells (SFCs) in the VLP+ Archaeosome immunized mice was also higher than that of the other groups immunized with either VLP-free archaeosomes or VLPs formulated with complete/incomplete Freund's adjuvants. These results propound M. smithii archaeosomes-entrapped mzNL4-3 VLPs as a promising immunogen which specifically induces and augments T-helper 1 oriented responses against HIV antigens.

Lamivudine-PEGylated chitosan: a novel effective nanosized antiretroviral agent.

Due to the fact that a definite treatment for AIDS disease has not been discovered so far, antiretroviral drugs are relatively significant in controlling the disease. In this study, Lamivudine- which is an old and effective anti-AIDS drug- was connected to PEGylated chitosan nanoparticle in order to produce a new and greater version of Lamivudine. First, physicochemical studies such as HNMR, FTIR spectroscopy and CHN analysis were performed to ensure the proper synthesis. Following that, Lamivudine-PEGylated Chitosan (LPC) drug was evaluated in terms of its inhibitory capability in producing HIV virions and its cytotoxicity in different doses. HIV virions were created by transfection of psPAX2, PmzNL4-3 and pMD2.G plasmids into HEK293T cell line. Also, assessment of the P24 amounts of cell supernatant was performed using ELISA method. Among the different doses of LPC drug (0.1, 1, 10 and 100µM), it was found that 0.1µM was the most effective and least toxic dose compared to Lamivudine in the same dose. Results of this study indicate that LPC drug has the ability to inhibit HIV virus replication in a more significant way in comparison to the old drug. Besides, the drug has a low cytotoxicity and is effective with a lower dose.