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.

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).

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.

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.

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.

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.

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.

Association of interleukin-18 gene promoter polymorphisms with breast cancer.

Interleukin-18 [IL-18] gene promoter polymorphism is reported to be a genetic risk factor for several types of cancer. The aims of this investigation were to evaluate and compare the frequencies of IL-18 gene promoter polymorphisms at positions -137 [G/C] and -607 [C/A] in breast cancer patients and healthy controls as well as to study the contribution of these data with clinicopathological parameters at diagnosis. The studied populations comprised 250 cases with breast carcinoma and 206 healthy subjects. IL-18 gene promoter polymorphisms at positions -137 and -607 were amplified in patient and control groups using allele specific polymerase chain reaction [AS-PCR]. The frequencies of GG, GC and CC genotypes of -137 SNP were 141 [56.4%], 96 [38.4%] and 13 [5.2%] in patients vs. 110 [53.4%], 72 [34.9%] and 24 [11.7%] in controls, respectively. A significant decrease of the CC genotype was observed in patients [p = 0.04]. The frequency of the CC genotype at position -137 was also significantly higher in patients with metastasis than non-metastatic patients [21.4% vs. 4.3%] [p = 0.02]. There was no significant association between genotype frequencies at position -607 with breast cancer or its clinicopathological parameters at diagnosis. Moreover, allelic frequencies at these positions did not contribute to breast cancer incidence. The distribution of IL-18 gene haplotypes and genotype combinations were not significantly different between patients and normal control individuals. This is the first report investigating the contribution of IL-18 gene promoter polymorphisms to breast cancer. These results suggest contrast effects of IL-18 gene in cancer induction and progression. Key words: Breast cancer, IL-18, polymorphism.

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.

Distinguishing between primary infection and reinfection with rubella vaccine virus by IgG avidity assay in pregnant women.

During the mass measles/rubella vaccination campaign in 2003 in Iran, many pregnant women were vaccinated mistakenly or became pregnant within 1 month of vaccination. To distinguish pregnant women who were affected by rubella vaccine as primary infection from those who had rubella reinfection from the vaccine, serum samples were collected 1-3 months after the campaign from 812 pregnant women. IgG avidity assay showed that 0.3% of the women had no rubella-specific IgG response; 14.4% had low-avidity anti-rubella IgG and were therefore not immune to rubella before vaccination; 85.3% had high-avidity anti-rubella IgG and were regarded as cases of reinfection.

Rubella serology in pregnant women attending health centres of Tehran University of Medical Sciences.

This study was performed in 2003-05 to determine the serological status of a sample of pregnant women as a preliminary study for the rubella vaccination programme. Out of 965 pregnant women attending health centres affiliated to Tehran University of Medical Sciences for prenatal care, the estimated rubella immunity rate was 91.1% (95% CI: 89.3%-92.9%) and the nonimmunity rate was 8.9% (95% CI: 7.1%-10.7%). The rubella immunity rate differed in different areas of Tehran but not significantly so. However, there was a significant difference in the level of rubella immunity by the number of persons per household and by age, but no significant relationship with economic status, occupation or level of education.

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.

Characterization of mutations in the VP(1) region of Sabin strain type 1 polioviruses isolated from vaccine-associated paralytic poliomyelitis cases in Iran.

BACKGROUND: The live-attenuated oral polio vaccine used to interrupt poliovirus transmission is genetically unstable. Reversion of some attenuating mutations, which normally occurs during vaccine strain replication in some recipients, and can rarely cause vaccine-associated paralytic poliomyelitis (VAPP). The poliovirus eradication program designed by the World Health Organization (WHO) includes immunization with OPV in addition to careful surveillance of all acute-flaccid paralysis (AFP) cases. OBJECTIVES: In Iran we last isolated imported wild poliovirus in 2000 and the immunization coverage was 100% in 2002. During 2001, there were three AFP cases with residual paralysis from which Sabin-like type 1 polioviruses were isolated in our national polio laboratory. STUDY DESIGN: The complete VP(1) region of the three isolates was sequenced and amino acid substitutions associated with these neurovirulent isolates were recorded. RESULTS: These isolates had either 4, 2 or 1 nucleotide substitution(s) in the VP(1) region, corresponding to amino acid change in the VP(1) of isolate 1 of either (H-[149]->Y), (T-[106]->A) or (I-[90]->L), respectively. CONCLUSIONS: Surveillance of the VAPP cases in countries where endemic transmission has recently ceased increases our understanding of the important neurovirulent mutations in vaccine-strain isolates and assists in planning the next step in the eradication program in these countries.

Association of chemokine receptor 5 (CCR5) delta32 mutation with Behçet's disease is dependent on gender in Iranian patients.

OBJECTIVE: Behçet's disease (BD) is a recurrent multi-system inflammatory disorder caused by the combinations of multiple genetic and environmental factors. CCR5 is a Th1-dominant chemokine receptor whose levels are increased in patients with active BD. It is believed that a 32 bp deletion in the CCR5 gene reduces the expression of this receptor on the cell surface. The aim of the present study was to investigate the association of CCR5 delta32 allele with BD in Iranian patients. METHODS: The study included 100 patients with BD and 380 healthy controls. Polymerase chain reaction (PCR) amplification was used for analysis of CCR5 delta32 allele. RESULTS: The frequency of CCR5 delta32 allele was not statistically different between 100 patients with BD and 380 healthy individuals. However, categorizing patients according to gender revealed a significant difference in distribution of the CCR5 delta32 allele in female patients compared with female control individuals (p = 0.047, fisher's exact test, OR = 2.66). CONCLUSION: The results suggest that the CCR5 delta32 allele may be a genetic risk factor for BD in Iranian women. These results warrant further investigation to clarify the underlying mechanism of CCR5 deficiency in the initiation of BD.

Distinguishing between primary measles infection and vaccine failure reinfection by IgG avidity assay.

In this study in the Islamic Republic of Iran 365 measles cases were evaluated to distinguish between primary infection with measles and reinfection due to secondary vaccine failure. All cases previously confirmed by detection of specific IgM were tested for IgG avidity. A secondary immune response was seen in 18.4% of patients. All unvaccinated patients (16.7%) showed a primary immune response. Of 244 patients with documented vaccination, 75.8% showed a primary immune response and 24.2% showed a secondary immune response, thereby indicating a secondary vaccine failure. Almost all measles reinfections (99%) were seen in patients >10 years old, indicating that vaccination for 10-year-old children is recommended.

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.