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.

Sustainable yarn production using leftover fabric from apparel industries.

A huge amount of waste was generated from the apparel industries. This study aims to develop the process of producing recycled yarn from apparel waste. The apparel leftover fabric was converted to fiber, and the fiber was mixed with virgin cotton in different ratios to produce sustainable 6/1 Ne rotor yarn. The produced yarn qualities viz. count strength product (CSP), elongation percentage, total quality index (TQI) and tenacity were decreased linearly, and opposite scenario observed for thick and thin places, neps, imperfection index (IPI) and hairiness (H) attributes with increasing the amount of waste addition with virgin cotton. The leftover fabric (LOF) can be utilized to develop a sustainable yarn and to zero waste management.

A LATS biosensor screen identifies VEGFR as a regulator of the Hippo pathway in angiogenesis.

The Hippo pathway is a central regulator of tissue development and homeostasis, and has been reported to have a role during vascular development. Here we develop a bioluminescence-based biosensor that monitors the activity of the Hippo core component LATS kinase. Using this biosensor and a library of small molecule kinase inhibitors, we perform a screen for kinases modulating LATS activity and identify VEGFR as an upstream regulator of the Hippo pathway. We find that VEGFR activation by VEGF triggers PI3K/MAPK signaling, which subsequently inhibits LATS and activates the Hippo effectors YAP and TAZ. We further show that the Hippo pathway is a critical mediator of VEGF-induced angiogenesis and tumor vasculogenic mimicry. Thus, our work offers a biosensor tool for the study of the Hippo pathway and suggests a role for Hippo signaling in regulating blood vessel formation in physiological and pathological settings.

In vitro expression of HPV16 E7 linked to HMGB1 immunoadjuvant in mammalian cells.

BACKGROUND: E7 is the major transforming protein of human papillomavirus (HPV) that plays important role in maintaining the proliferative state in HPV-infected cells. Furthermore, high mobility group 1 protein (HMGB1) is a highly conserved component of chromatin that can be secreted by macrophages and activated monocytes and thus functions as an inflammation mediator. METHODS: In the current study, cloning of HMGB1 gene and also HPV16E7-HMGB1 was performed in pEGFP-N1 eukaryotic expression vector in order to evaluate their expression in mammalian cells. For this purpose, the HEK-293T cells were transfected by pEGFP-E7, pEGFP-HMGB1 and pEGFP-E7-HMGB1 using TurboFect delivery system. The levels of protein expression were assessed by flow cytometry and fluorescent microscopy at 48 hr after transfection, as well as by western blot analysis using anti-GFP polyclonal antibody. RESULTS: Our data showed a clear band of ~ 684 bp and ~ 981 bp related to HMGB1 and E7-HMGB1 genes in agarose gel, respectively. The expression of HMGB1-GFP and E7-HMGB1-GFP proteins was confirmed for the bands of ~ 53 kDa and ~ 64 kDa in the transfected cells using western blot analysis, respectively. The linkage of HMGB1 gene to E7 could likely neutralize the negative charges of E7, thus a clear band of 64 kDa was detected instead of 76 kDa in western blot analysis. Moreover, the percentage of expression for E7-GFP, HMGB1-GFP and E7-HMGB1-GFP was 76 %, 55 %, and 52 %, in comparison with pEGFP-N1 (~82 %) as a positive control. Indeed, HMGB1 linked to HPV16 E7 gene decreased transfection efficiency of E7 DNA in HEK-293T cells. CONCLUSION: Generally, the electrophoretic mobility of HPV16 E7 was changed due to the linkage of HMGB1 gene. Furthermore, the fusion protein could be efficiently expressed in mammalian cells for the next use in immunotherapy (Fig. 3, Ref. 51).

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.

Estimation of influenza and severe acute respiratory illness incidence (burden) in three provinces of the Islamic Republic of Iran, 2012 and 2013.

There are few estimates of influenza burden in the WHO Region for the Eastern Mediterranean. In this study we estimated the burden of severe acute respiratory infection (SARI) and influenza-associated SARI (F-SARI) in selected provinces of Islamic Republic of Iran, the trends of SARI and confirmed cases of influenza (F-SARI) over 12 months (seasonality), and the age groups most at risk. Using the electronic Iranian influenza surveillance system and data of cases in sentinel hospitals of 3 selected provinces, we estimated the monthly trend (seasonality) of incidence for SARI and F-SARI, overall incidence of SARI and F-SARI and their disaggregation by age with the aid using the Monte Carlo technique. The age groups most at-risk were children aged under 2 years and adults older than 50 years.

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.

Estimation of influenza and severe acute respiratory illness incidence [burden] in three provinces of the Islamic Republic of Iran, 2012 and 2013

There are few estimates of influenza burden in the WHO Region for the Eastern Mediterranean. In this study we estimated the burden of severe acute respiratory infection [SARI] and influenza-associated SARI [F-SARI] in selected provinces of Islamic Republic of Iran, the trends of SARI and confirmed cases of influenza [F-SARI] over 12 months [seasonality], and the age groups most at risk. Using the electronic Iranian influenza surveillance system and data of cases in sentinel hospitals of 3 selected provinces, we estimated the monthly trend [seasonality] of incidence for SARI and F-SARI, overall incidence of SARI and F-SARI and their disaggregation by age with the aid using the Monte Carlo technique. The age groups most at-risk were children aged under 2 years and adults older than 50 years

Peu d'estimations sont disponibles sur la charge de la grippe dans la Région OMS de la Méditerranée orientale. Dans la présente étude, nous avons estimé la charge de morbidité due aux infections respiratoires aiguës sévères [IRAS] et aux IRAS associés à la grippe dans certaines provinces de la République islamique d'Iran, ainsi que les tendances des IRAS et des cas de grippe confirmés sur 12 mois [saisonnalité], et les groupes d'âge les plus exposés. En utilisant le système électronique iranien de surveillance de la grippe et les données relatives aux cas des hôpitaux sentinelles de trois provinces sélectionnées, nous avons estimé la tendance mensuelle [saisonnalité] de l'incidence des IRAS et des IRAS dus à la grippe, l'incidence globale des IRAS et des IRAS dues à la grippe et leur ventilation par âge grâce à la méthode de Monte Carlo. Les groupes d'âge les plus exposés au risque étaient les enfants de moins de 2 ans et les adultes de plus de 50 ans

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.

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

كثيراً ما توجد تباينات بين تغطية التطعيم المسجلة وبين المستوى الحقيقي للمناعة في المجتمع. ولتقدير تغطية التلقيح ضد الحصبة في جنوب شرق جمهورية إيران الإسلامية تم إجراء دراسة مقطعية في 3 مناطق خلال صيف عام 2011.فتم اختيار عينة عنقودية من 1368 طفلاً تتراوح أعمارهم ما بين 30-54 شهراً باستخدام الاحتمال المتناسب مع الحجم. وتم فحص عينات مصلية من 663 منهم - ممن تلقوا حقنتين من اللقاح الثلاثي ضد الحصبة والنكاف والحصبة الألمانية [MMR]- للتأكد من وجود الجلوبيولين المناعي G [IgG]المضاد للحصبة. لقد بلغت تغطية التطعيم بالجرعة الثانية من لقاح MMR 93.7%.وكان انتشار الجلوبيولين المناعي G المضاد للحصبة لدى أولئك الذين تلقوا جرعتين على الأقل من لقاح MMR 94.6%.وكان هناك ارتباط ذو دلالة إحصائية بين النتائج السيرولوجية وبين المتغيرات التي تعكس ضعف إمكانية الوصول إلى الخدمات الصحية. وبجمع النتائج السيرولوجية مع بيانات التغطية قُدِّرت نسبة الجمهور المحمي ضد الحصبة بـ 88.6%، والتي كانت أقل من الحدود المعينة لأهداف التخلص من الحصبة

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 MMRvaccine 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 pooraccessibility 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.

Il existe souvent des écarts entre la couverture vaccinale enregistrée et le niveau réel d'immunité d'une communauté donnée. Afin d'estimer la couverture vaccinale antirougeoleuse dans le sud-est de la République islamique d'Iran, une étude transversale a été menée dans trois districts durant l'été 2011. En appliquantl'échantillonnage en grappes avec probabilité proportionnelle à la taille, 1368 enfants âgés de 30 à 54 mois ont été sélectionnés. Des prélèvements de sérum de 663 enfants de l'échantillonnage ayant reçu deux injections du vaccincontre la rougeole, les oreillons et la rubéole (ROR) ont été analysés à la recherche d'anticorps IgG antirougeoleux. La couverture vaccinale pour la deuxième dose de vaccin ROR était de 93,7 %. La prévalence des anticorps IgG antirougeoleux chez les enfants ayant reçu au moins deux doses du vaccin ROR était de 94,6 %. Il existait une association statistiquement significative entre les résultats sérologiques et les variables qui reflétaient l'accès insuffisant aux services de santé. En combinant les résultats sérologiques et les données de couverture, la proportion de la communauté protégée contre la rougeole a été estimée à 88,6 %, soit un taux inférieur aux seuils définis pour les objectifs d'élimination de la rougeole.

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.

Learning chronobiology by improving Wikipedia.

Although chronobiology is of growing interest to scientists, physicians, and the general public, access to recent discoveries and historical perspectives is limited. Wikipedia is an online, user-written encyclopedia that could enhance public access to current understanding in chronobiology. However, Wikipedia is lacking important information and is not universally trusted. Here, 46 students in a university course edited Wikipedia to enhance public access to important discoveries in chronobiology. Students worked for an average of 9 h each to evaluate the primary literature and available Wikipedia information, nominated sites for editing, and, after voting, edited the 15 Wikipedia pages they determined to be highest priorities. This assignment (http://www.nslc.wustl.edu/courses/Bio4030/wikipedia_project.html) was easy to implement, required relatively short time commitments from the professor and students, and had measurable impacts on Wikipedia and the students. Students created 3 new Wikipedia sites, edited 12 additional sites, and cited 347 peer-reviewed articles. The targeted sites all became top hits in online search engines. Because their writing was and will be read by a worldwide audience, students found the experience rewarding. Students reported significantly increased comfort with reading, critiquing, and summarizing primary literature and benefited from seeing their work edited by other scientists and editors of Wikipedia. We conclude that, in a short project, students can assist in making chronobiology widely accessible and learn from the editorial process.

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.

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.

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.

Sequence and phylogenetic analyses of neuraminidase gene of Iranian seasonal influenza H1N1 viruses from 2005-2009 and corresponding vaccine strains.

Prophylaxis of influenza A virus infections is based on the vaccines inducing antibodies to the major viral antigens, hemagglutinin (HA) and neuraminidase (NA). Since these antigens continuously change during virus replication in various hosts, only the currently circulating strains should be used in the vaccines. Besides, monitoring of the naturally occurring changes in HA, NA, and respective genes, especially those associated with resistance to the NA inhibitors is necessary. The NA genes of 30 Iranian isolates of influenza H1N1 virus from the seasons 2005-2009 were sequenced and subjected to the sequence and phylogenetic analyses. The seasonal isolates turned out to be closely related to the corresponding vaccine strains, except for the 2007-2008 isolates, which also displayed a higher nucleotide variation. A resistance to the NA inhibitors was found in the 2008-2009 isolates only. The average nucleotide identities of the isolates with corresponding vaccine strains for the years 2005-2009 were 98.83%, 98.55%, 98.7%, 97.55%, and 98.76%, respectively.