Currently available anti-influenza drugs are often associated with limitations such as toxicity and the appearance of drug-resistant strains. Therefore, there is a pressing need for the development ...of novel, safe and more efficient antiviral agents. In this study, we evaluated the antiviral activity of zinc oxide nanoparticles (ZnO-NPs) and PEGylated zinc oxide nanoparticles against H1N1 influenza virus.
The nanoparticles were characterized using the inductively coupled plasma mass spectrometry, x-ray diffraction analysis, and electron microscopy. MTT assay was applied to assess the cytotoxicity of the nanoparticles, and anti-influenza activity was determined by TCID50 and quantitative Real-Time PCR assays. To study the inhibitory impact of nanoparticles on the expression of viral antigens, an indirect immunofluorescence assay was also performed.
Post-exposure of influenza virus with PEGylated ZnO-NPs and bare ZnO-NPs at the highest non-toxic concentrations could be led to 2.8 and 1.2 log10 TCID50 reduction in virus titer when compared to the virus control, respectively (P < 0.0001). At the highest non-toxic concentrations, the PEGylated and unPEGylated ZnO-NPs led to inhibition rates of 94.6 and 52.2%, respectively, which were calculated based on the viral loads. There was a substantial decrease in fluorescence emission intensity in viral-infected cell treated with PEGylated ZnO-NPs compared to the positive control.
Taken together, our study indicated that PEGylated ZnO-NPs could be a novel, effective, and promising antiviral agent against H1N1 influenza virus infection, and future studies can be designed to explore the exact antiviral mechanism of these nanoparticles.
Human papillomavirus (HPV) is the most common sexually transmitted infection worldwide, affecting about 80% of women up to the age of 50. The persistent infection of high risk-HPV types (HR-HPV) is ...the leading cause of cervical cancer, the fourth most common cancer of women. Therefore, we aimed to evaluate the frequency and typing of HPV in the genital lesions in the Iranian population.
This descriptive-analytic study was conducted on a population in the South-Khorasan province of Iran. All of the participants were sexually active and were checked for evident cervical warts. Biopsy samples were collected from various lesions, and all samples were tested for detection and genotyping of HPV using a reverse dot blot hybridization method (HPV direct flow CHIP).
In overall, 370 samples were evaluated; 10 cases (2.7%) were male and the rest were female. The mean age of patients was 33.3 ± 8.5 years, of which 48.1% were in the age range from 25 to 36 years. Among the samples, 345 (93.2%) were positive for HPV-DNA; the low risk HPV types (LR-HPV) and HR-HPV were identified among 80.9% and 15.5% of tissue samples, respectively. Among the LR-HPV, HPV-6, 11, 42 and 54 were the most common genotypes, and HPV-16 and 39 were prevalent HR-HPV types detected. The number of pregnancies, marriage age, and partner infection were not significantly related to the HPV types. Types 42 had a declining pattern toward aging, and HPV-11 was increasing toward aging.
The number of samples with HR-HPV was rather high. Due to the greater frequency of infection in the age range of 25-35 years, it is advised that all individuals referred to gynecological clinics at gestational age be tested for HPV types.
Abstract
Hepatitis B virus (HBV), along with Hepatitis C virus chronic infection, represents a major risk factor for hepatocellular carcinoma (HCC) development. However, molecular mechanisms involved ...in the development of HCC are not yet completely understood. Recent studies have indicated that mutations in
CTNNB1
gene encoding for β-catenin protein lead to aberrant activation of the Wnt/ β-catenin pathway. The mutations in turn activate several downstream genes, including
c-Myc
, promoting the neoplastic process. The present study evaluated the mutational profile of the
CTNNB1
gene and expression levels of
CTNNB1
and
c-Myc
genes in HBV-related HCC, as well as in cirrhotic and control tissues. Mutational analysis of the β-catenin gene and HBV genotyping were conducted by direct sequencing. Expression of β-catenin and
c-Myc
genes was assessed using real-time PCR. Among the HCC cases, 18.1% showed missense point mutation in exon 3 of
CTNNB1
, more frequently in codons 32, 33, 38 and 45. The frequency of mutation in the hotspots of exon 3 was significantly higher in non-viral HCCs (29.4%) rather than HBV-related cases (12.7%,
P
= 0.021). The expression of β-catenin and
c-Myc
genes was found upregulated in cirrhotic tissues in association with HBV infection. Mutations at both phosphorylation and neighboring sites were associated with increased activity of the Wnt pathway. The results demonstrated that mutated β-catenin caused activation of the Wnt pathway, but the rate of
CTNNB1
gene mutations was not related to HBV infection. HBV factors may deregulate the Wnt pathway by causing epigenetic alterations in the HBV-related HCC.
Hepatitis B virus, with 8 known distinct genotypes, is one of the most serious health problems which results to liver injuries. The surface gene of Hepatitis B virus completely overlaps with the ...polymerase gene. Mutations in the RT gene result in changes in the overlapping hepatitis B surface antigen.
The present study aimed to evaluate the genotypes and prevalence of mutations in a segment of S and RT gene in HBV isolates in Southern Khorasan, Iran.
This was a population-based study comprising 5,235 randomized samples for HBV screening. A nested-polymerase chain reaction (PCR) test was followed by direct sequencing, and the sequences blast with present sequences of NCBI database for genotyping. Alignment and phylogenic analysis was performed using MEGA-6 software, and mutation pattern of this segment was finally surveyed in Bioedit software.
The mean age was 39.07 ± 14.04 years, with 52.2% female and 47.8% male. All isolates belonged to HBV genotype D, sub-genotype D1. The most amino acid substitutions of surface protein were Q129H (34.42%) and A168V (8.2%), other escape mutants observed in this study were P127L/T, S117G, T125M, S143L, D144E and E164D. In the RT gene, Q149K was the most frequently identified amino acid substitution (9.83%), followed by L122F (8.19%), N118D/T (6.55%), L157M (4.91%), and H124Y (3.27%).
This finding represents an ongoing dominancy of HBV genotype D in Eastern Iran, corresponding to other parts of Iran. There were a lot of variations in the S gene leading to an escape mutation, some of which affected the corresponding area of the RT region.
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.
Chlamydia trachomatis (CT), the most common bacterial sexually transmitted infection (STI), leads to pelvic inflammatory disease, infertility and chronic pelvic pain in women as well as an increased ...risk of vertical transmission, conjunctivitis and pneumonitis in infants. It may also be a co-factor along with human papillomavirus (HPV) in cervical cancer progression. We aimed to determine the prevalence of CT genotypes in genital specimens of women from South Khorasan, Iran and to test the association between CT and cytology statistics.
This was a cross-sectional study on 248 Pap smear samples from women who visited a gynecologist for routine Pap smear testing in South Khorasan province. Nested polymerase chain reaction (PCR) was used to test the residual fluids of Pap smears for CT-DNA after cytological examination. Direct sequencing, alignment and phylogenic analyses were performed on eight samples to identify their genotypes.
The mean age of patients was 37.54 ± 5.21 years. Most samples had a normal cytology (214 cases, 86.29%). Overall, 31 samples were positive for CT infection (12.5%) of which 20 (9.34%) were normal and 11 (32.35%) were abnormal, with the frequency difference being significant (P=0.022). The co-infection of CT/HPV in total was identified in 14 cases (5.6%). The results of sequencing eight samples out of the 31 CT positive samples revealed the detection of genotypes D and E, each with four cases.
We show that a high prevalence of genital CT infection is present in women with both normal and abnormal cytology; however, the higher prevalence among women in the abnormal group may indicate its involvement in cervical neoplasia.
Background: Anelloviridae is a viral family which is considered as a constant component of human virome. Given the ubiquitous nature of the virus infection and the long-standing relationship between ...the virus and the host, in the present study, we aimed at investigating the presence of Anelloviruses in the urine samples of children in a cross-sectional study. Materials and Methods: The urine samples of 50 children who were referred to Hazrat Ali Asghar Children's Hospital, affiliated to Iran University of Medical Sciences, Tehran, Iran, were obtained. Three TaqMan real-time polymerase chain reactions (PCRs) were carried out for Anellovirus detection. A phylogenetic tree was drawn for positive products after PCR amplification, purification, and nucleotide sequencing. SPSS, version 20, was used for statistical analyses. Results: Children's mean age ± standard deviation was 4.30 ± 1.47 years and 56% (28/50) were female. Real-time PCR revealed that Anellovirus was positive in 12% (6/50). Furthermore, PCR-sequencing results showed that torque teno virus was detected in 83.3% (5/6) and SEN virus in 16.6% (1/6) of the Anellovirus positive samples. In addition, 86% (5/6) of the children with positive samples were female. No significant difference was detected between any of the demographic characteristics and Anellovirus positivity (P > 0.05). Conclusion: According to our preliminary study, the presence of Anelloviruses in the urine samples of asymptomatic children in Iran is striking, although limited sample size and age range limitations might have affected the comprehensive results of our study.
Human parvovirus B19 (B19V) is one of the smallest DNA viruses and shows great resistance to most disinfectants. Therefore, it is one of the common contaminant pathogens present in blood and plasma ...products. Parvovirus 4 (PARV4) is a newly identified parvovirus, which is also prevalent in parenteral transmission. In this study, we aimed to evaluate the prevalence of B19V and PARV4 DNA among patients with hemophilia in Birjand County in eastern Iran.
This was a cross-sectional epidemiological study comprising nearly all people with hemophilia in this region. Whole blood samples were taken after patient registration and sent for plasma isolation. After nucleic acid extraction, B19V was detected with real-time polymerase chain reaction, PARV4 DNA was then detected using sensitive semi-nested PCR.
In total, there were 86 patients with hemophilia, with mean age 28.5±1.5 years. Of these, 90.7% were men and 9.3% women; 84.9% had hemophilia A and 7.0% had hemophilia B. We found 11 patients (12.8%) were positive for B19V DNA and 8 were positive (9.3%) for PARV4 DNA. The prevalence of B19V was higher in middle-aged groups rather than younger people, whereas PARV4 infection was more common in younger patients (
<0.05).
There was a high prevalence of B19V and PARV4 infection in this high-risk group of patients with hemophilia. Due to the clinical significance of the B19 virus, imposing more precautionary measures for serum and blood products is recommended.
Objectives
Features of occult hepatitis B virus (HBV) infection among the anti‐hepatitis B core antigen (anti‐HBc) positives have yet to be described in more details. This study aimed to determine ...the molecular prevalence of occult HBV infection (OBI), and association to risk factors among seropositives for anti‐HBc.
Methods
This was part of a community‐based screening project that included 5234 cases. All participants completed a questionnaire on demographic and socio‐epidemiological information. Then, the blood samples were collected and tested for anti‐HBc and HBsAg using ELISA method. To identify OBI, nested‐polymerase chain reaction (PCR) assays were performed for HBV‐S and X genes, and viral load was determined using an in‐house real‐time PCR. Sequencing and phylogenetic analysis have been implemented for genotyping.
Results
Overall, 596 cases, positive only for anti‐HBc were included in the study. OBI was detected among 61 cases (10.2%). The genotype and subgenotype of HBV among all of them was D1, except one that was D4. Most of them had low viral loads ranged from 1.2 × 102 to 1.34 × 10
3 copies/mL; 19.6% had undetectable viral loads. Important mutations in surface protein and reverse transcriptase were sI92T, sQ129H, rtL80I, rtS85F, rtL91I. The prevalence of OBI was related to some risk factors, such as tattooing (P = 0.02), sexual activities (P = 0.009), and diabetes (P = 0.031).
Conclusion
Our study suggests that OBI should be considered among anti‐HBc seropositive subjects. This form of HBV infection was accompanied with some mutations, risk factors, and diseases. However, further investigations are needed to determine virological importance of documented mutations.
Background:
Patients with chronic kidney failure and those undergoing chronic hemodialysis (CHD) treatment are at high risk of infection with hepatitis C virus (HCV). The incidence of occult HCV ...infection (OCI) in CHD remains controversial and the real burden of HCV in this population may be affected by the rate of OCI. This study evaluates the molecular assessment of OCI in CHD in an Iranian population.
Methods:
All subjects on CHD in the South Khorasan province of Iran were invited for participation in the study. Whole blood samples were taken and serological, clinical, and demographic information was recorded. HCV-RNAs were checked in serum and peripheral blood mononuclear cells (PBMCs) using an in-house semi-nested PCR assay. Viral load was determined using a real-time PCR-based quantification kit. Sequencing was performed to determine genotypes.
Results:
Overall, 120 cases participated in the study; 57.5% were male and the rest were female. In serum samples, no positive case was found for HCV-RNA. In PBMC samples, 2/120 (1.6%) were positive for HCV-RNA (95% CI, 0.002 to 0.059); the mean age of OCI positive cases was 37.5 ± 19.2 years which was significantly lower than OCI negative cases (P = 0.026). Only one case had detectable viral load which was 49 IU/mL. The only HCV genotype identified was 1a.
Conclusion:
This study showed that there is a risk of OCI among CHD patients; the very low and undetectable viral loads of OCI warrant further follow-up molecular testing for earlier diagnosis and treatment in the era of DAA.
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