Diverse Expression Patterns of EBV Oncogenes (LMP2A, EBV-Encoded microRNA, and EBV-encoded dUTPase) in EBV Associated Gastric Carcinoma and their Association with Viral Loads.

The chromogenic in situ hybridization (CISH) test is the gold standard for detecting Epstein-Barr virus (EBV)-associated gastric carcinoma (GC). Real-time (RT) PCR method is also a sensitive test that can detect the viral load in samples. As such, three EBV oncogenes were investigated in this study. RNA extraction and cDNA synthesis were performed on GC tissues of nine patients, who were previously confirmed to have EBVGC subtype. In addition, 44 patients that had positive RT-PCR but negative CISH results were also included as the control group. TaqMan RT-PCR analysis was performed to determine the expression of EBV-encoded microRNAs, and the expression of EBV-encoded dUTPase, as well as LMP2A, was analyzed by SYBR Green RT-PCR. EBV-encoded microRNAs and LMP2A were identified in 2 out of 9 (22%) EBVGC subtypes. In addition, EBV-encoded dUTPase was detected in 4 out of 9 (44.5%) EBVGC subtypes. EBV-encoded dUTPase was also expressed in a sample of the control group. The expression of LMP2A, EBV-encoded microRNAs, and EBV-encoded dUTPase viral oncogenes in patients with high EBV viral loads indicates that these expressions correlate with viral loads. Our findings indicate that the EBV-encoded dUTPase gene may have a role in EBVGC patients' non-response to treatment and might be considered a Biomarker-targeted therapy.

Cdc6 expression is induced by HPV16 E6 and E7 oncogenes and represses E-cadherin expression.

Cervical cancer is one of the most common cancers in women worldwide, and its development is related to two viral oncoproteins E6 and E7 from high-risk human papillomaviruses. Aberrant expression of E-cadherin is associated with epithelial-to-mesenchymal transition (EMT), and it is frequently seen in cervical cancer. However, the underlying mechanisms involved in E-cadherin suppression in cervical cancer are not clear. We studied the effects of human papillomavirus 16 (HPV16) E6 and E7 on E-cadherin and Cdc6 (cell division cycle 6) expression in the HCT-116 cell line. We also assessed the relationship between Cdc6 and E-cadherin expression in cells expressing HPV16 E6 and E7 proteins. The results showed that HPV16 E6 and E7 proteins reduce E-cadherin expression, and HPV16 E6-expressing cells undergo a more profound suppression of E-cadherin compared with cells expressing HPV16 E7. Our results also revealed that HPV16 E6 and E7 oncoproteins induce Cdc6 expression, whereas suppression of Cdc6 protein by short hairpin RNA restores E-cadherin expression. Induction of Cdc6 expression in HCT-116 cells was greater with E6 than with E7, a finding that was consistent with the corresponding changes in E-cadherin expression. These observations suggest that Cdc6 overexpression is an important factor for E-cadherin reduction in cells expressing HPV16 E6 and E7 proteins and may have an important role in the metastasis of HPV-associated cancers.Cancer Gene Therapy advance online publication, 11 November 2016; doi:10.1038/cgt.2016.51.

The effect of oxamflatin on the E-cadherin expression in gastric cancer cell line.

Gastric cancer is among the leading causes of cancer-related death, and the symptoms are commonly characterized in advanced stages. Histone acetylation is among the most important epigenetic alterations occurring during cancer development. In addition, reduced E-cadherin expression is a major contributor in the process of tumor cell invasion and metastasis. Oxamflatin is a histone deacetylase inhibitor that has been suggested as a promising anti-tumor agent; yet its effect on the viability and invasion of gastric tumor cells is unclear. We aimed to assess the impact of oxamflatin on the viability of gastric tumor cells and expression of E-cadherin as a marker of tumor invasion susceptibility. In this study, MKN-45 cells were treated with 1, 2.5 and 5 mM oxamflatin and followed by MTT assay after 24-48 h of incubation. To determine E-cadherin expression in treated cells, total RNA was extracted and reverse transcribed to complementary DNA, followed by quantitative real-time PCR. MTT results showed that the viability of MKN-45 cells declines with increasing concentrations of oxamflatin. The results of quantitative real-time PCR showed increased expression of E-cadherin following treatment with oxamflatin at the concentration of 2.5 mM compared with 1 mM. The present results showed that oxamflatin can induce E-cadherin expression and also reduce cell viability in the MKN-45 cell line. On the basis of these findings, oxamflatin can be further considered for the prevention of tumor metastasis.

Measles vaccination coverage and seroprevalence of anti-measles antibody in south-east Islamic Republic of Iran.

Discrepancies often exist between recorded immunization coverage and the real immunity level in a community. To estimate the vaccination coverage against measles in south-east Islamic Republic of Iran, a crosssectional study was conducted in 3 districts during summer 2011. Using probability proportional to size cluster sampling, 1368 children aged 30-54 months were selected. Serum samples of 663 who had received 2 injections of mumpsmeasles- rubella (MMR) vaccine were checked for anti-measles IgG. Vaccination coverage for the second dose of MMR vaccine was 93.7%. The prevalence of anti-measles IgG in those who had received at least 2 MMR vaccine doses was 94.6%. There was a statistically significant association between the serological results and variables that reflected poor accessibility to health services. Combining serological results with coverage data, the proportion of the community protected against measles was estimated as 88.6%, which was below the limits defined for the measles elimination goals.

Human parvovirus B19: a review.

UNLABELLED: Parvovirus B19 (B19V) is a small non-enveloped single-stranded DNA (ssDNA) virus of the family Parvoviridae, the subfamily Parvovirinae, the genus Erythrovirus and Human parvovirus B19 type species. It is a common community-acquired respiratory pathogen without ethnic, socioeconomic, gender, age or geographic boundaries. Moreover, the epidemiological and ecological relationships between human parvovirus B19, man and environment have aroused increasing interest in this virus. B19V infection is associated with a wide spectrum of clinical manifestations, some of which were well established and some are still controversial, however, it is also underestimated from a clinical perspective. B19V targets the erythroid progenitors in the bone marrow by binding to the glycosphingolipid globoside (Gb4), leading to large receptor-induced structural changes triggering cell death either by lysis or by apoptosis mediated by the nonstructural (NS)1 protein. The pattern of genetic evolution, its peculiar properties and functional profile, the characteristics of its narrow tropism and restricted replication, its complex relationship with the host and its ample pathogenetic potential are all topics that are far from a comprehensive understanding. The lack of efficient adaptation to in vitro cellular cultures and the absence of animal models have limited classical virological studies and made studies on B19V dependent on molecular biology. The present review looks at the nature of this virus with the view to provide more information about its biology, which may be useful to the present and future researchers. KEYWORDS: human parvovirus B19; respiratory pathogen; biology; genome; fifth disease; transient aplastic crisis; anemia.

Establishment of a standard seed lot system of an Iranian Mumps virus strain; RS-12, for mass production of mumps and MMR vaccines.

BACKGROUND: At present the mumps virus strain used for production of mumps vaccine for our local use is Hoshino strain. However, according to our National public health policies, this strain should be replaced with a safer strain. Based on our previous data, the Iranian mumps strain; RS-12 has been proved to be the most suitable alternative to Hoshino strain with little or no adverse events following vaccination METHODS: The aim of the present study was to optimize propagation of RS-12 strain and prepare standard seeds for vaccine mass production. The virus was inoculated to cells using different methods and different multiplicity of infection (MOI). The viral suspensions were harvested using different methods. Quality control tests were run at different stages. RESULTS: Maximum viral yield was achieved when cell suspensions were inoculated at MOI of 1:10 and incubated at 36-37ºC for 48 hours, followed by replacement of the media and incubation at 33-34 ºC for 5-7 days. Filtration did not affect the viral titre. A standard seed lot system was successfully established and experimental batches of MMR vaccines were produced. CONCLUSION: The established seed lot system has met all requirements of WHO regulations and could be used in mass production of safe and efficacious mumps and MMR vaccines. Clinical trials are in progress for this newly produced vaccine.

Efficacy of influenza vaccination in patients with cirrhosis and inactive carriers of hepatitis B virus infection.

BACKGROUND: Influenza can causes morbidity and mortality that are greatly enhanced in patients with underlying chronic diseases such as Cirrhotic patients. This study was performed to assess the immunogenicity of Influenza vaccination in patients with cirrhosis and inactive carriers of Hepatitis B virus infection. METHODS: In this clinical study 93 enrolled subjects divided into 3 groups: Cirrhotic (N=28), Inactive carriers of Hepatitis B (N=31) and subjects (N=34). All the participants were vaccinated by Influenza vaccine (Influvac®). Serum samples were taken before and 4 weeks after vaccination and the Humoral Immunogenicity was assessed by the Hemagglutination Inhibition (HI) test. RESULTS: Four weeks after vaccination, seroconversion rates of vaccine strains ranged between 71.4% and 100% in Group 1, 70.6% and 94.1% in Group 2, and 58.1% and 80.7% in Group 3. No significant differences were seen in the rates of Seroconversion and antibody Geometric Mean Titers (GMTs) against Influenza A (H1N1 and H3N2) vaccine components in the three groups (P>0.05).The rates of Seroconversion and antibody GMTs against Influenza B vaccine component were significantly higher in Cirrhotic and inactive carriers of Hepatitis B than healthy subjects (P<0.005). No significant (P>0.05) differences in the rates of Seroprotection were observed within the three groups. Antibody GMTs against all three strains of Influenza vaccine increased significantly (P<0.001) after vaccination in three groups. CONCLUSION: Influenza vaccination is effective in Cirrhotic patients and inactive carriers of Hepatitis B as well as healthy individuals. It means that vaccination should be considered in such patients in order to reduce the morbidity and mortality of Influenza.

A comparative analysis of routine techniques: Reverse transcriptase polymerase chain reaction (RT-PCR) and five cell lines for detection of enteroviruses in stool specimens.

BACKGROUND AND OBJECTIVES: Each year, Enteroviruses infect millions of people and cause different diseases. The agents are usually detected using cell culture. RD (Rhabdomyosarcoma) and L20B (L cells) are among the recommended cells by the World Health Organisation (WHO) for this purpose. Even though cell culture is the most common method used in diagnosing Enteroviruses in stool specimens, this particular method poses some problems, which include false positive or negative results, lack of a unique cell line for diagnosing all Enterovirus types in addition to being time consuming. For these reasons, an attempt was made to find better techniques of Enterovirus detection. RT-PCR (Reverse Transcriptase Polymerase Chain Reaction) is a technique used in place of the cell culture method. In this study, the cell culture method was compared with RT-PCR for detection of Enteroviruses in stool specimens. MATERIAL AND METHOD: First, the chloroform treated stool samples were inoculated onto five cell lines, including RD, L20B, Hep-2 (Human Epidermoid carcinoma cell line), Vero (Verde Reno) and GMK (Green Monkey Kidney). The results were then compared with data from Enterovirus detection using the RT-PCR technique. RESULTS AND CONCLUSION: The difference between RT-PCR and cell culture results was significant. Enteroviruses were detected in 24% of specimens using RT-PCR while cell lines could isolate Enteroviruses in just 14.4% of the samples.

Molecular Epidemiology of Measles Virus before and after the 2003 Mass Vaccination Campaign for Measles/Rubella in Iran.

BACKGROUND: Molecular epidemiology of measles virus (MV) is important, not only to measure the success of measles vaccination programs but also to monitor the circulation and elimination of the virus worldwide. In this study, we compared MV obtained from patients before the 2003 mass vaccination MR campaign and viruses detected after 2003 until 2008 in Iran. METHODS: The nucleoprotein (N) gene of 29 MV strains circulating in Iran between 2002 and 2008 were amplified by RT-PCR and subjected to sequence and phylogenetic analysis. RESULTS: Molecular characterization of MV studied here revealed that although the outbreaks in Iran were associated with MV genotype D4, the isolated viruses clearly belonged to several different lineages. Maximum and minimum homology within the 29 Iranian strains in our study was100% and 94.9% within the carboxyl terminus of the N gene, respectively. Using ClustalX program, the alignment of Iranian MV sequences showed nine lineages. CONCLUSION: This study provides the usefulness of MV sequence analysis for the demonstration of local interruption of indigenous strain transmission as well as providing a valuable means for monitoring the elimination processes of MV control.

Analysis of the hemagglutinin and neuraminidase genes of human Influenza A/H3N2 viruses circulating in iran between 2005 and 2007: antigenic and phylogenetic relationships to vaccine strains.

OBJECTIVES: To study the antigenic variations in influenza A/H3N2 viruses circulating in Iran for characterization and phylogenetic relationships to vaccine strains. METHODS: RT-PCR, full sequencing of hemagglutinin (HA) and neuraminidase (NA) genes and analysis by sequence handling and phylogenetic programs were done. RESULTS: The HA sequences of 2007 isolates fell within the clade represented by the HA of A/Brisbane/10/07 and characterized by the amino acid changes relative to the HA of A/Wisconsin/67/05, G50E and K140I. The only isolate in 2006 fell within A/Berlin/02/06 with V112I and K173E changes. The 2005 isolates characterized by Y159F, S189N and S227P changes within A/California/07/04. In all isolates we had E190D which is important because this was responsible for the loss of ability of A/H3N2 viruses to bind to chicken red blood cells. There were some substitutions in the antigenic sites of the HA. Similar to other studies, conserved residues for catalytic sites and also framework sites of NA supporting the catalytic residues were detected. We had some changes in the variable regions of the NA head domain. CONCLUSION: Comparison between Iranian viruses and vaccine strains showed high similarity between them and vaccine strains used in the northern hemisphere.

First report of respiratory syncytial virus and human metapneumovirus co-infection in a 2-year-old kawasaki patient in iran.

BACKGROUND: Respiratory virus infections in children are a leading cause of morbidity and mortality worldwide. METHODS: A total of 897 clinical specimens were collected from February 2007 to January 2008 and transported to the National Influenza Center. Two hundred and two samples belonged to children under the age of six from 897 specimens, described above, were selected. Then they were tested for influenza virus types and subtypes by real time PCR assay subsequently, the specimens were tested for RSV and hMPV by hemi-nested multiplex PCR and parainfluenza viruses type 1-4 by hemi-nested multiplex PCR and adenovirus by hemi-nested PCR. RESULTS: The throat swab was taken from the Kawasaki case with the history of chicken's contact. The specimen was tested for all influenza subtypes especially H5N1 and the results were negative. Meanwhile PCR was done for screening of other respiratory viruses that results came out positive for RSV and hMPV. CONCLUSION: In the present study, we demonstrated the possibility to detect dual infection caused by RSV and hMPV, but because of the extravagant pattern of this case, more investigation is suggested specially on Kawasaki patients.

Amantadine-resistant influenza A (H3N2) viruses in Iran.

Adamantanes have been used for the prophylaxis and treatment of Influenza A virus (IAV) infections worldwide. However, they have limited use because of increasing number of resistant viruses during recent years. In investigating the frequency of amantadine-resistant IAVs (H3N2) circulating in Iran in 2005-2008, we found that M2 sequences of recently circulating viruses that were amantadine-resistant contained a Ser31Asn mutation. Thus, adamantanes should not be used for treatment or prophylaxis of recent IAVs (H3N2) infections. In future, their potential use will depend on the resistance of circulating viruses.

A new core promoter mutation and premature stop codon in the S gene in HBV strains from Iranian patients with cirrhosis.

In order to define hepatitis B virus (HBV) mutational patterns in Iran, nucleotide sequences obtained from 91 patients and encompassing the precore, basal core promoter (BCP) and surface (S) regions, were compared. The patients were grouped as asymptomatic carriers, chronic active hepatitis or cirrhotic patients. Genotypes and mutations were determined by sequencing and phylogenetic analysis. All strains belonged to genotype D, and most of them to subgenotype D1. All but two strains specified ayw2, one ayw3 and one adw2 determinants. Two deletions of 8- or 20-bp were found in the X region in eight strains, six from patients with chronic active hepatitis. Eight of 21 strains from patients with cirrhosis harboured unusual mutations such as a stop codon at position 69 in the S region or a previously not described mutation in the BCP region ((1761)TC/ATTTG(1766)). All patients infected by strains with the stop codon mutation had detectable HBsAg and high viral load. The accumulation of mutations found in the BCP and S regions in HBV strains from patients with chronic active hepatitis and cirrhosis may predict disease progression in Iranian HBsAg carriers.

Influenza surveillance in the Islamic Republic of Iran from 1991 to 2001.

To better understand the annual distribution of influenza virus in our country, we isolated and typed 45 viruses from 1043 patients with acute respiratory illnesses in a 10-year study conducted by the National Influenza Centre of the Islamic Republic of Iran. The seasonal distribution of influenza typically ran from November to April. Type A influenza was most common during the winters of 1991-92, 1997-98 and 2000-01 and type B influenza was most common during 1992-5 and 1996-97. Both type A and type B viruses circulated in 1995-96 and 1998-2000. A serological survey based on haemagglutination inhibition test confirmed our findings. The annual pattern of strains isolated was similar to the worldwide pattern during the same interval.

Influenza surveillance in the Islamic Republic of Iran from 1991 to 2001

To better underst and the annual distribution of influenza virus in our country, we isolated and typed 45 viruses from 1043 patients with acute respiratory illnesses in a 10-year study conducted by the National Influenza Centre of the Islamic Republic of Iran. The seasonal distribution of influenza typically ran from November to April. Type A influenza was most common during the winters of 1991-92, 1997-98 and 2000-01 and type B influenza was most common during 1992-5 and 1996-97. Both type A and type B viruses circulated in 1995-96 and 1998-2000. A serological survey based on haemagglutination inhibition test confirmed our findings. The annual pattern of strains isolated was similar to the worldwide pattern during the same interval