Aida Gholoobi - Academia.edu (original) (raw)

Papers by Aida Gholoobi

Research paper thumbnail of Temozolomide and Doxorubicin Combined Treatment by Graphene Oxide-Cyclodextrin Nanocarriers for Synergistic Inhibition of Glioblastoma Cells

BioNanoScience, Jan 17, 2024

Research paper thumbnail of Association Between Serum Levels of Anti-heat Shock Protein 27 Antibody and Liver Cell Injury in Chronic Hepatitis B

Indian Journal of Clinical Biochemistry, Jan 31, 2023

Background Heat Shock Protein 27 (HSP27), an anti-HBV factor, exists in the intracellular and ext... more Background Heat Shock Protein 27 (HSP27), an anti-HBV factor, exists in the intracellular and extracellular spaces. Serum HSP27 (sHSP27) is an in ammatory modulator and is associated with elevated pro-in ammatory cytokines and with a higher likelihood of hepatocellular carcinoma. SHSP27 results in natural antibody production (anti-HSP27-Ab) that is more stable and easily detectable compared to sHSP27. We aimed to investigate any potential association between anti-HSP27-Ab level and chronic hepatitis B (CHB) progression and in ammation indicated by liver cell injury and HBV replication. Methods This cross-sectional study was conducted on 91 patients with CHB and 92 individuals without CHB. Following demographic data collection, anti-HSP27-Ab, serum lipids including total cholesterol, triglyceride, LDL-C, HDL-C, and aminotransferase levels were measured using enzymatic assays in participants' serum samples. HBV DNA was also measured by quantitative PCR in CHB patients. Results Bivariate and multivariate analyses showed a signi cantly higher mean level of anti-HSP27-Ab in CHB than in healthy individuals (0.304 vs. 0.256AU/ml, P-value = 0.015). These levels held signi cant differences in the CHB subgroups of male patients, at the age of 50 years and above, non-smokers, patients with elevated aminotransferase levels, and hypotriglyceridemia (P-value < 0.05). However, no difference was found between the antibody levels and HBV DNA copies (P-value > 0.05). Conclusion This study provides evidence that anti-HSP27 antibody levels can re ect the degree of liver necrosis indicated by aminotransferase levels. Regarding the higher incidence rate of HBV-associated complications in 50 to 60-year-old men, monitoring the antibody can be bene cial in managing this group of CHB patients, which deserves further investigation.

Research paper thumbnail of Association Between Serum Levels of Anti-Heat Shock Protein-27 Antibody and Liver Cell Injury in Chronic Hepatitis B

Research Square (Research Square), Dec 29, 2022

Background Heat Shock Protein 27 (HSP27), an anti-HBV factor, exists in the intracellular and ext... more Background Heat Shock Protein 27 (HSP27), an anti-HBV factor, exists in the intracellular and extracellular spaces. Serum HSP27 (sHSP27) is an in ammatory modulator and is associated with elevated pro-in ammatory cytokines and with a higher likelihood of hepatocellular carcinoma. SHSP27 results in natural antibody production (anti-HSP27-Ab) that is more stable and easily detectable compared to sHSP27. We aimed to investigate any potential association between anti-HSP27-Ab level and chronic hepatitis B (CHB) progression and in ammation indicated by liver cell injury and HBV replication. Methods This cross-sectional study was conducted on 91 patients with CHB and 92 individuals without CHB. Following demographic data collection, anti-HSP27-Ab, serum lipids including total cholesterol, triglyceride, LDL-C, HDL-C, and aminotransferase levels were measured using enzymatic assays in participants' serum samples. HBV DNA was also measured by quantitative PCR in CHB patients. Results Bivariate and multivariate analyses showed a signi cantly higher mean level of anti-HSP27-Ab in CHB than in healthy individuals (0.304 vs. 0.256AU/ml, P-value = 0.015). These levels held signi cant differences in the CHB subgroups of male patients, at the age of 50 years and above, non-smokers, patients with elevated aminotransferase levels, and hypotriglyceridemia (P-value < 0.05). However, no difference was found between the antibody levels and HBV DNA copies (P-value > 0.05). Conclusion This study provides evidence that anti-HSP27 antibody levels can re ect the degree of liver necrosis indicated by aminotransferase levels. Regarding the higher incidence rate of HBV-associated complications in 50 to 60-year-old men, monitoring the antibody can be bene cial in managing this group of CHB patients, which deserves further investigation.

Research paper thumbnail of Design and Construction of a Eukaryotic Cloning Vector Encoding the mpt51 Gene of Mycobacterium tuberculosis

PubMed, Apr 1, 2019

Background: Tuberculosis (TB) is the leading cause of death by infectious diseases worldwide, and... more Background: Tuberculosis (TB) is the leading cause of death by infectious diseases worldwide, and especially prevalent in developing countries. Several vaccines against TB have been developed, recently. The aim of the present study was to design and construct a cloning vector encoding Mycobacterium tuberculosis (MTB) mpt51 gene. Methods: DNA was extracted from MTB H37Rv strain. Gene-specific primers were designed using Gene Runner software and the mpt51 gene was amplified by PCR. The amplified fragment and pcDNA3.1(+) cloning vector were both digested with restriction enzymes, the mpt51 fragment was ligated into the vector, and the Escherichia coli (E. coli) TOP10 strain were transformed by the recombinant plasmid. Positive clones were identified by colony PCR, restriction enzyme digestion, and DNA sequencing. Results: The mpt51 gene was successfully cloned into pcDNA3.1(+). A 6400 bp band for the pcDNA3.1(+)/mpt51 recombinant plasmid and a 926 bp band for mpt51 were observed by colony PCR, and restriction enzyme digestion on agarose gels. The DNA sequence was 100% homologous with the mpt51 fragment of H37Rv in GenBank. Conclusion: In the current study, the mpt51 gene of MTB was correctly cloned into pcDNA3.1(+). The expression of this recombinant vector can be studied in eukaryotic cells. Moreover, it is possible to determine the efficacy of this vector as a DNA vaccine candidate, and to test its protective function compared to BCG in animal models in future.

Research paper thumbnail of Evaluation of autophagy induction and inhibition in the Huh7.5 cell line through flow cytometry

Medical laboratory sciences, 2017

Background: Autophagy is a physiologic process in which double membrane vesicles engulf damaged p... more Background: Autophagy is a physiologic process in which double membrane vesicles engulf damaged proteins and organelles for delivering them to lysosome in order to degrade and recycle them via lysosomal digestion. Beclin1 is one of the basic proteins involved in the initial step of auto phagosome formation. In the current study, the effects of exogenous Beclin1 and 3-methyladenine (3-MA) to induce and inhibit of autophagy were assessed in Huh7.5 cells as an in vitro models of hepatocellular carcinoma. Materials and Methods: The recombinant pcDNA-Beclin1was transfected into Huh7.5 cells. Also, the cytotoxicity of 3-methyladenine (3-MA) were determined in Huh7.5 cells and the cells were treated with 3-methyladenine (3-MA). The autophagy induction and inhibition was conducted via LC3 staining as a main autophagy marker using flowcytometry. Results: The result of this study suggest that the over expression ofexogenousBeclin1 in Huh7.5 cells elevated the autophagosome formation as shown by intracellular autophagosomal marker LC3-II staining for about 32.32 % and 3-MA decreased it up to2% compared with control cells in which the stained LC3-II was12.08.The IC50 of 3-methyladenine (3-MA) was 7.2 mM in Huh7.5 cells. Conclusion: Recombinant beclin1 may be used as a potential autophagy inducer agent and3methyladenine (3-MA) inhibits autophagy formation in Huh7.5 cell. The staining autophagy formation marker LC3-II with specific antibody is a reliable method to measure autophagy activation via flow cytometry.

Research paper thumbnail of Isolation, cloning and molecular analysis of ag85a and tb10.4 genes from Mycobacterium tuberculosis

Medical laboratory sciences, Oct 16, 2017

Background: Novel tuberculosis (TB) vaccines that aim to boost and/or replace Bacillus Calmette-G... more Background: Novel tuberculosis (TB) vaccines that aim to boost and/or replace Bacillus Calmette-Guerin (BCG) are currently in development. DNA vaccines can stimulate both humoral and cell-mediated immunity in different animal models of TB and is thought to be a promising strategy in the development of new vaccines against TB. The aim of this study was to design and construct a DNA vaccine encoding ag85a and tb10.4 fusion genes of Mycobacterium tuberculosis. Materials and Methods: tb10.4 fragment was amplified by PCR and the product was digested with restriction enzymes. Next, it was cloned into the pcDNA3.1+ plasmid. The ag85a gene and pcDNA3.1+/tb10.4 plasmid were digested by EcoRI and BamHI restriction enzymes. Constructed vector was sequenced. The molecular analysis was done using bioinformatics software. New chimeric vector containing ag85a-tb10.4 genes were purified. Expression of pcDNA3.1+/tb10.4-ag85a plasmid was confirmed in eukaryotic cells. Results: Fragments of 297 bp for tb10.4 and 1017 bp for ag85a were observed in agarose gel electrophoresis. Alignment of ag85a-tb10.4 genome sequence with reference genes in GenBank showed exact identities that indicate correction of all cloning procedures. Transfection of eukaryotic cells with pcDNA3.1+/tb10.4-ag85a vector and existence of tb10.4-ag85a fusion gene were both confirmed with RT-PCR. Conclusion: In this study, tb10.4 and ag85a genes were isolated from Mycobacterium tuberculosis H37Rv strain and cloned into pcDNA3.1+. Also, the capability of constructed vector in producing fusion ag85a-tb10.4 protein was confirmed with RT-PCR. pcDNA3.1+/tb10.4-ag85a vector can be used for further studies in future.

Research paper thumbnail of Comparison of the culture and PCR methods for diagnosis of Mycobacterium tuberculosis in different clinical specimens

Clinical Biochemistry, Sep 1, 2011

Background: Tuberculosis remains a global epidemic, especially in developing countries, including... more Background: Tuberculosis remains a global epidemic, especially in developing countries, including Iran. Rapid diagnosis of active Mycobacterium tuberculosis infection plays a critical role in controlling the spread of tuberculosis. Conventional methods may take up to several weeks or longer to produce results. In addition to multiplicity of steps involved in conventional detection, including isolation, identification and drug susceptibility testing, the slow growth rate of M. tuberculosis is also responsible for this lengthy time. Objectives: The aim of this study was to compare the polymerase chain reaction (PCR) and culture methods for the detection of M. tuberculosis in different clinical specimens. Materials and Methods: This study was performed on different samples (urine, gastric aspirate, bronchoalveolar lavage, pleural fluid, cerebrospinal fluid, ascetic fluid and joint fluid specimens) of tuberculosis suspected patients. M. tuberculosis DNA was extracted directly from different samples using two different protocols. Next, PCR was performed using three sets of specific primers to detect members of Mycobacterium genus, M. tuberculosis complex and non-tuberculosis Mycobacteria. The results were then compared with that of the culture method, which is considered as the gold standard method. Results: The concordance rate between the three sets of primers was calculated and IS6110/buffer PCR method showed good agreement with the LJ culture method (κ = 0.627, P < 0.0001). The sensitivity of IS6110/buffer PCR was 58.33%, with specificity of 77.78%; the positive and negative predictive values were 100% and 78.26%, respectively. Buffer method for DNA extraction was proved to give a higher accuracy to PCR in comparison with the boiling method. Conclusions: PCR method is a valuable, cost-effective and alternative tool for quick diagnosis of active tuberculosis in different clinical specimens.

Research paper thumbnail of Relationship between the prevalence and severity of non‐alcoholic fatty liver disease and coronary artery disease: Findings from a cross‐sectional study of a referral center in northeast Iran

JGH open, Apr 27, 2022

Background and AimNon‐alcoholic fatty liver disease (NAFLD) is becoming increasingly prevalent wo... more Background and AimNon‐alcoholic fatty liver disease (NAFLD) is becoming increasingly prevalent worldwide, and cardiovascular diseases are the most common cause of death in NAFLD patients. The present study aimed to evaluate the possible relationship between the presence and severity of NAFLD and coronary artery disease (CAD).MethodsA cross‐sectional study was conducted on 296 patients (122 men and 174 women, with mean age 54.10 ± 9.33 years) referred to the catheterization laboratory of Imam Reza Hospital affiliated to the Mashhad University of Medical Sciences, Mashhad, Iran, for elective coronary angiography to investigate the presence and severity of CAD. Additionally, all patients underwent abdominal ultrasonography (USG) to detect NAFLD and its severity.ResultsAmong the 296 patients, 187 (63.2%) had CAD and 160 (50.1%) had NAFLD. NAFLD patients had significantly higher prevalence of obesity (odds ratio [OR] = 1.047, 95% confidence interval [CI] = 1.002–1.094), hypertension (OR = 1.909, 95% CI = 1.027–3.55), hyperlipidemia (OR = 3.474, 95% CI = 1.862–6.482), and CAD (OR = 2.009, 95% CI = 1.100–3.669). The percentage of patients with normal vessels was higher in the non‐NAFLD group, followed by the group with mild and severe NAFLD (P < 0.001). However, single‐ and multi‐vessel disease incidences among the non‐NAFLD, mild, and severe NAFLD groups were 36.1, 43.1, and 63.7%, respectively. Interestingly, the percentage of patients with two‐vessel stenosis was significantly higher in severe NAFLD patients than mild and non‐NAFLD patients (P < 0.001).ConclusionThe prevalence and severity of NAFLD were independently associated with CAD. Mild NAFLD was primarily observed among patients with normal and non‐obstructive coronary artery patients, while severe NAFLD was more frequent in extensive CAD patients with multi‐vessel disease.

Research paper thumbnail of A case of recurrent meningitis caused by drug resistant Enterococcus fecalis in a patient with ventriculoperitoneal shunt

KAUMS Journal, Jul 15, 2007

ABSTRACT Background: Iatrogenic meningitis is an important complication in neurosurgery. This inf... more ABSTRACT Background: Iatrogenic meningitis is an important complication in neurosurgery. This infection is not common. This paper presents the first report of recurrent shunt meningitis caused by drug resistant Enterococcus fecalis in Iran. Case report: We report a case with recurrent shunt meningitis caused by Enteroccocus fecalis in a 9-year old girl with ventriculoperitoneal shunt which did not recover by antibiotic therapy until the removal of the infected shunt. Discussion and Conclusion: Patients with hydrocephalus who are treated with ventriculoperitoneal shunts are susceptible to bacterial infections. The rate of this nosocomial infection varies from 2% to 22%. It is suggested to pay more attention to symptoms of CNS infections among patients with cerebrospinal fluid drainage. Keywords: Recurrent meningitis, Ventriculoperitoneal shunt, Enteroccocus fecalis, Drug resistant,

Research paper thumbnail of Designing a label‐free electrochemical aptasensor based on polypyrrole‐<scp>l</scp>‐cysteine‐reduced graphene oxide nanocomposite for detection of 25‐hydroxyvitamin D<sub>3</sub>

Biotechnology and Applied Biochemistry, Jun 27, 2023

Research paper thumbnail of Synthesis of γ-Fe2O3 Nanoparticles Capped with Oleic Acid and their Magnetic Characterization

Iranian Journal of Science and Technology Transaction A-science, Feb 22, 2017

In recent years, superparamagnetic iron oxide nanoparticles have attracted a great attention due ... more In recent years, superparamagnetic iron oxide nanoparticles have attracted a great attention due to their various biomedical applications, such as magnetic resonance imaging, targeted drug delivery, and hyperthermia. In this article, c-Fe 2 O 3 magnetic nanoparticles (Maghemite) were prepared in oleic acid media by co-precipitation method. The oleic acid, a monounsaturated fatty acid was used as the capping and stabilizing agent during the synthesis of the magnetic nanoparticles. Characterization of obtained nanoparticles were performed using powder X-ray diffraction (PXRD), field emission scanning electron microscopy (FESEM), Fourier transform infrared spectra (FTIR), and vibrating sample magnetometer (VSM). The crystallite size of c-Fe 2 O 3 nanoparticles was achieved in the range between 16.2 and 26.8 nm. The FESEM demonstrated the regular spheres of c-Fe 2 O 3 nanoparticles. The obtained nanoparticles were coated with oleic acid indicating by FTIR analysis. The resulted oleic acid-coated nanoparticles were shown superparamagnetic properties (*52 emu/g). This suggested method is simple and rapid to fabricate superparamagnetic nanoparticles which make them appropriate candidates for theranostic application in future studies.

Research paper thumbnail of Artemisia Absinthium Extract Attenuates the Quinolinic Acid-Induced Cell Injury in OLN-93 Cells

Current Drug Discovery Technologies

Objective: Increased quinolinic acid (QA) accumulation has been found in many neurodegenerative d... more Objective: Increased quinolinic acid (QA) accumulation has been found in many neurodegenerative diseases. Artemisia absinthium (A. absinthium) has been reported to have neuroprotective and antioxidant activities. This study was designed to evaluate the effect of A. absinthium in QAinduced neurotoxicity in OLN-93 Cells. Methods: OLN-93 cells were cultured in a DMEM medium containing 10% (v/v) fetal bovine serum, 100 units/ml penicillin, and 100 μg/ml streptomycin. The cells were pretreated with concentrations of A. absinthium extract for two h and then exposed to QA for 24 h. After 24 h cell viability, the level of malondialdehyde (MDA), reactive oxygen species (ROS), and apoptotic cells were quantitated in OLN-93 Cells. Results: Pretreatment with A. absinthium extract prevented the loss of cell viability in OLN-93 cells. ROS generation, lipid peroxidation, and apoptosis in QA -injured OLN-93 cells were reduced following A. absinthium extract pretreatment. Conclusion: A. absinthium e...

Research paper thumbnail of Relevance of aptamers as targeting ligands for anticancer therapies

Research paper thumbnail of Seroprevalence of Herpes Simplex Viruses Types 1 and 2 in a Population, Age 15-35 Years, of Mashhad City

Iranian biomedical journal, Mar 1, 2023

Background: Considering the high prevalence and clinical importance of herpes simplex virus (HSV)... more Background: Considering the high prevalence and clinical importance of herpes simplex virus (HSV) infection worldwide, we aimed to evaluate the seroprevalence of HSV-1 and HSV-2 in a population aged between 15 and 35 years in Mashhad, Iran. Methods: This cross-sectional study was conducted on 916 cases composed of 288 (31.4%) men and 628 (68.6%) women. Using ELISA method, the presence of IgM and IgG antibodies against HSV-1 and HSV-2 was assessed. Results: Among the population studied, 681 (74.3%) cases were positive for anti-HSV antibodies, while 235 (25.7%) cases were negative. Moreover, no IgMs were found and all positive subjects had IgG antibodies. Age (p < 0.001), occupation (p < 0.001), education (p = 0.006), smoking (p = 0.029), and BMI (p = 0.004) demonstrated a significant association with HSV-1 and HSV-2 infection. Conclusion: Our study indicates a high seroprevalence of HSV infection; however, there was no cases positive for IgM antibodies, suggesting the high prevalence of latent infection.

Research paper thumbnail of Herpes Simplex Infection in Cardiovascular Disease (CVD) Patients: A Seroepidemiological Study

Journal of Kermanshah University of Medical Sciences, Mar 14, 2023

Background: Herpes simplex viruses have been implicated as a cause of cardiovascular disease (CVD... more Background: Herpes simplex viruses have been implicated as a cause of cardiovascular disease (CVD) due to their widespread distribution and ability to infect human vascular endothelial cells. Objectives: The present study aimed to evaluate the link between Herpes simplex infections and cardiovascular disease. Methods: This case-control study involved 236 patients aged 35-65, 118 with known cardiovascular disease and 118 controls. Patients' cardiovascular disease evaluation was based on data from questionnaires, a specialist's physical examinations, and paraclinical tests. Herpes simplex viruses (HSV)-antibodies (IgM, IgG) were determined using ELISA in serum samples, and the biochemical blood indicators were analyzed to examine their relationship to the level of HSV antibodies. All the data were analyzed using SPSS software version 20. Results: IgM antibodies against herpes simplex were negative among all 236 patients. The positivity rate of IgG antibodies against herpes simplex was 58.8 and 51.2% in the case and control groups, respectively with no significant difference (P = 0.253). Patients with cardiovascular disease had a 0.587-fold higher positive rate of IgG antibodies than patients in the control group (odds ratio (OR) = 0.587). The mean age of patients with positive IgG antibodies was significantly lower than patients with negative IgG antibodies (P = 0.012). IgG positivity among men and women (P = 0.670) and different job statuses (P = 0.866) were not significantly different. Conclusions: The positivity rate of IgG antibodies against herpes simplex was not significantly different among the study groups. Although patients with positive IgG antibodies' mean age was lower than those with negative IgG antibodies, the gender and job status were not different.

Research paper thumbnail of A new DNA vaccine expressing HspX-PPE44-EsxV fusion antigens of Mycobacterium tuberculosis induced strong immune responses

PubMed, Jul 1, 2020

Objectives: Infection with tuberculosis (TB) is regarded as a major health issue. Due to the emer... more Objectives: Infection with tuberculosis (TB) is regarded as a major health issue. Due to the emergence of antibiotic resistance during TB treatment, prevention via vaccination is one of the most effective ways of controlling the infection. DNA vaccines are developed at a greater pace due to their ability in generating a long-lasting immune response, higher safety compared to the live vaccines, and relatively lower cost of production. In the present study, we evaluated a new DNA vaccine encoding the fusion HspX-PPE44-EsxV antigens, separately, and in combination with Bacillus Calmette-Guérin (BCG) administration, in a prime-boost method in mice. Materials and methods: A novel DNA vaccine encoding HspX-PPE44-EsxV fusion antigen of Mycobacterium tuberculosis was constructed, and RT-PCR and Western blot analysis were performed to verify the expression of the antigen. Female BALB/c mice were divided into five groups (PBS, BCG, pcDNA3.1 (+) vector, pDNA/HspX-PPE44-EsxV vaccine, and the BCG-prime boost groups). In order to evaluate the immunogenicity of the recombinant vector, BALB/c mice were injected with 100 μg of pDNA at 2-week intervals. Then, cytokine assay was conducted using eBioscience ELISA kits (Ebioscience, AUT) according to manufacturers' instructions to evaluate the concentrations of IL-4, IL-12, TGF-β, and IFN-γ. Results: The concentrations of INF-γ, IL-12, and TGF-beta were significantly increased compared to the control groups (P<0.001). INF-γ and IL-12 production were increased significantly in pDNA/HspX-PPE44-EsxV+BCG group compared to pDNA/HspX-PPE44-EsxV group (P<0.001). Conclusion: This study showed that the present DNA vaccine could induce a high level of specific cytokines in mice. It was also shown that using this DNA vaccine in a BCG prime-boost protocol can produce significant amounts of IFN-γ, IL-12, and TGF-β.

Research paper thumbnail of Designing and construction a DNA vaccine encoding the fusion fragment of cfp10 and ag85a immunodominant genes of Mycobacterium tuberculosis

Medical laboratory sciences, May 21, 2017

Background: Pathogenic mycobacteria are one of major causes of human morbidity and mortality. Myc... more Background: Pathogenic mycobacteria are one of major causes of human morbidity and mortality. Mycobacterium tuberculosis (M. tuberculosis) is an etiological agent of human tuberculosis. Designing new vaccines including DNA vaccines may be considered as new approaches for preventing of TB. Materials and Methods: M. tuberculosis H37Rv was grown on Lowenstein Jensen medium for 4 weeks at 37ºC and then DNA was extracted. The cfp10 gene was amplified by PCR. After digesting the PCR product and the plasmid, cfp10 fragment was ligated into the vector using T4 DNA ligase. Then, Ag85A was subcloned into pcDNA/cfp10. Escherichia coli strain JM109 bacteria were transformed by the desired construct. Clone confirmations were performed by colony PCR, restriction enzyme digestion and DNA sequencing. Recombinant vector was transfected into HeLa cells and total RNA was extracted, then cDNA was synthesized using oligo-dT. Finally PCR was performed by cfp10 primers. Results: The cfp10 was amplified by PCR method and the PCR products were visualized by agarose gel electrophoresis. The cfp10 fragments showed 303 bp in length. The cfp10 cloned into pcDNA. Then, Ag85Awas ligated into pcDNA/cfp10 after digestion correctly. Colony-PCR and restriction enzyme digestion and sequencing confirmed the cloning the fusion Ag85A/cfp10 fragment. Finally, after cDNA synthesis, expression of vector was confirmed in eukaryotic system. Conclusion: Cloning of Ag85A/cfp10 genes of M. tuberculosis were performed correctly. It can use as a DNA vaccine for investigation the immune responses in animal models in future studies.

Research paper thumbnail of CTLA-4 Gene Haplotypes and the Risk of Chronic Hepatitis C Infection; a Case Control Study

PubMed, Oct 1, 2017

Background: The prevalence of hepatitis C virus (HCV) infection is increasing worldwide. Cytotoxi... more Background: The prevalence of hepatitis C virus (HCV) infection is increasing worldwide. Cytotoxic Tlymphocyte-associated protein 4 (CTLA-4) may play a role in the intensity of the disease. The aim of this study was to evaluate the association between genetic variants of the CTLA-4 and HCV infection. Methods: Restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) was performed as the genotyping assay at four different positions (+49 A>G, -318 C>T, -1722 T>C, and - 1661 A>G). Haplotypes were analyzed using PHASE software. Sixty-five HCV patients and 65 healthy individuals as controls who were referred to the hepatitis clinic in Mashhad, Iran, were recruited. Genomic DNA was extracted from whole blood of participants. Results: In a dominant analysis model of the -1661 position (GG vs. AA+AG), the AA genotype was more common in controls than in patients (adjusted P = 0.0003; OR = 0.15, 95% CI = 0.051 -0.42). The GCAT haplotype was also more prevalent in controls than in patients (adjusted P = 0.01; OR = 0.40, 95% CI = 0.20-0.81). Furthermore, the ACGT/ACGT diplotype was more common in controls than in patients (P = 0.0037; OR = 0.15, 95% CI = 0.04-0.54). In addition, the ACGT/ACAT diplotype was more frequent in patients than controls (adjusted P =0.003; OR = 2.48, 95% CI = 1.37- 4.50). Conclusion: Our results indicated that polymorphisms in CTLA-4 and certain haplotypes may affect the risk of HCV infection in our population, although a larger sample size may be required to confirm this association.

Research paper thumbnail of The Antiviral Effects of Curcumin Nanomicelles on the Attachment and Entry of Hepatitis C Virus

Iranian Journal of Virology, Jun 15, 2017

Background and Aims: Hepatitis C virus (HCV) is a member of the Flaviviridae family, which causes... more Background and Aims: Hepatitis C virus (HCV) is a member of the Flaviviridae family, which causes approximately 500,000 deaths annually. HCV infection treatment is often associated with significant adverse effects. Curcumin is an active ingredient of turmeric which has therapeutic anti-inflammatory effects in many diseases including infectious ones. Although curcumin is not soluble in water, if it is synthesized in the form of nanomicelles, it will be water soluble and can be absorbed in the gastrointestinal tract (GI). In this study, the antiviral effects of curcumin nanomicelles were investigated on the attachment and entry of HCV particles. Materials and Methods: The cytotoxicity of curcumin nanomicelles was determined in Huh7.5 cells and their antiviral effects on the attachment and entry of HCV was investigated in a cell culture system. Results: Curcumin nanomicelles could decrease the viral load in the cell culture supernatants compared to virus control. Conclusions: According to the results of this research, we determined the antiviral effects of curcumin nanomicelles in the later stages of HCV replication.

Research paper thumbnail of Evaluation of Immune Responses to a DNA Vaccine Encoding Ag85a-Cfp10 Antigen of Mycobacterium tuberculosis in an Animal Model

Jundishapur Journal of Microbiology, Dec 16, 2018

Background: Many studies indicate that the Bacillus Calmette-Guérin (BCG) vaccine has low protect... more Background: Many studies indicate that the Bacillus Calmette-Guérin (BCG) vaccine has low protective efficacy, especially in endemic areas. There are several factors in this assessment, such as genetic diversity of BCG strains, pre-exposure to environmental mycobacteria, and variations in host immune responses. Currently, more than 200 new vaccine candidates have been proposed, such as recombinant BCG, DNA, and subunit vaccines. However, none of them are superior to BCG. Nevertheless, several approaches are considered to reduce the cases of tuberculosis infection. Objectives: The aim of the present study was to evaluate the capability of the Ag85a-cfp10 fusion protein as a new chimeric protein in stimulating immune responses. Methods: A DNA vaccine encoding Ag85a-cfp10 fusion protein was constructed in the previous study. The expression of Ag85a-cfp10 fusion protein in host cells was confirmed by the RT-PCR method. Six pathogen-free female mice were injected intramuscularly at a total concentration of 100 µg/mL three times at two-week intervals. The BCG and the control groups received BCG and PBS vaccines, respectively. One month after the final immunization, mice were killed and their splenocytes were cultured in RPMI medium supplemented with 1% antibiotics and 10% serum. Four cytokines including IL-4, IL-12, TGF-β, and IFN-γ were measured in the culture supernatant using the ELISA test. Results: RT-PCR analysis showed that Ag85a-cfp10 recombinant vector is able to replicate in eukaryotic cells and produce mRNA. The vaccinated groups were compared to the control group, showing induction of high levels of cytokine production. Conclusions: Some reports depicted that DNA vaccines are able to induce humoral and cellular immune responses both in animal models and humans. Therefore, in the current study, the immune response was induced in mice, which were inoculated with recombinant expression plasmid, pcDNA3.1 (+)-Ag85a-cfp10. We showed that this recombinant vector can stimulate mycobacterial specific modulating cytokines. Nonetheless, analysis appeared that this vaccine is unable to stimulate cell mediated immunity, however, still further studies are needed in future.

Research paper thumbnail of Temozolomide and Doxorubicin Combined Treatment by Graphene Oxide-Cyclodextrin Nanocarriers for Synergistic Inhibition of Glioblastoma Cells

BioNanoScience, Jan 17, 2024

Research paper thumbnail of Association Between Serum Levels of Anti-heat Shock Protein 27 Antibody and Liver Cell Injury in Chronic Hepatitis B

Indian Journal of Clinical Biochemistry, Jan 31, 2023

Background Heat Shock Protein 27 (HSP27), an anti-HBV factor, exists in the intracellular and ext... more Background Heat Shock Protein 27 (HSP27), an anti-HBV factor, exists in the intracellular and extracellular spaces. Serum HSP27 (sHSP27) is an in ammatory modulator and is associated with elevated pro-in ammatory cytokines and with a higher likelihood of hepatocellular carcinoma. SHSP27 results in natural antibody production (anti-HSP27-Ab) that is more stable and easily detectable compared to sHSP27. We aimed to investigate any potential association between anti-HSP27-Ab level and chronic hepatitis B (CHB) progression and in ammation indicated by liver cell injury and HBV replication. Methods This cross-sectional study was conducted on 91 patients with CHB and 92 individuals without CHB. Following demographic data collection, anti-HSP27-Ab, serum lipids including total cholesterol, triglyceride, LDL-C, HDL-C, and aminotransferase levels were measured using enzymatic assays in participants' serum samples. HBV DNA was also measured by quantitative PCR in CHB patients. Results Bivariate and multivariate analyses showed a signi cantly higher mean level of anti-HSP27-Ab in CHB than in healthy individuals (0.304 vs. 0.256AU/ml, P-value = 0.015). These levels held signi cant differences in the CHB subgroups of male patients, at the age of 50 years and above, non-smokers, patients with elevated aminotransferase levels, and hypotriglyceridemia (P-value < 0.05). However, no difference was found between the antibody levels and HBV DNA copies (P-value > 0.05). Conclusion This study provides evidence that anti-HSP27 antibody levels can re ect the degree of liver necrosis indicated by aminotransferase levels. Regarding the higher incidence rate of HBV-associated complications in 50 to 60-year-old men, monitoring the antibody can be bene cial in managing this group of CHB patients, which deserves further investigation.

Research paper thumbnail of Association Between Serum Levels of Anti-Heat Shock Protein-27 Antibody and Liver Cell Injury in Chronic Hepatitis B

Research Square (Research Square), Dec 29, 2022

Background Heat Shock Protein 27 (HSP27), an anti-HBV factor, exists in the intracellular and ext... more Background Heat Shock Protein 27 (HSP27), an anti-HBV factor, exists in the intracellular and extracellular spaces. Serum HSP27 (sHSP27) is an in ammatory modulator and is associated with elevated pro-in ammatory cytokines and with a higher likelihood of hepatocellular carcinoma. SHSP27 results in natural antibody production (anti-HSP27-Ab) that is more stable and easily detectable compared to sHSP27. We aimed to investigate any potential association between anti-HSP27-Ab level and chronic hepatitis B (CHB) progression and in ammation indicated by liver cell injury and HBV replication. Methods This cross-sectional study was conducted on 91 patients with CHB and 92 individuals without CHB. Following demographic data collection, anti-HSP27-Ab, serum lipids including total cholesterol, triglyceride, LDL-C, HDL-C, and aminotransferase levels were measured using enzymatic assays in participants' serum samples. HBV DNA was also measured by quantitative PCR in CHB patients. Results Bivariate and multivariate analyses showed a signi cantly higher mean level of anti-HSP27-Ab in CHB than in healthy individuals (0.304 vs. 0.256AU/ml, P-value = 0.015). These levels held signi cant differences in the CHB subgroups of male patients, at the age of 50 years and above, non-smokers, patients with elevated aminotransferase levels, and hypotriglyceridemia (P-value < 0.05). However, no difference was found between the antibody levels and HBV DNA copies (P-value > 0.05). Conclusion This study provides evidence that anti-HSP27 antibody levels can re ect the degree of liver necrosis indicated by aminotransferase levels. Regarding the higher incidence rate of HBV-associated complications in 50 to 60-year-old men, monitoring the antibody can be bene cial in managing this group of CHB patients, which deserves further investigation.

Research paper thumbnail of Design and Construction of a Eukaryotic Cloning Vector Encoding the mpt51 Gene of Mycobacterium tuberculosis

PubMed, Apr 1, 2019

Background: Tuberculosis (TB) is the leading cause of death by infectious diseases worldwide, and... more Background: Tuberculosis (TB) is the leading cause of death by infectious diseases worldwide, and especially prevalent in developing countries. Several vaccines against TB have been developed, recently. The aim of the present study was to design and construct a cloning vector encoding Mycobacterium tuberculosis (MTB) mpt51 gene. Methods: DNA was extracted from MTB H37Rv strain. Gene-specific primers were designed using Gene Runner software and the mpt51 gene was amplified by PCR. The amplified fragment and pcDNA3.1(+) cloning vector were both digested with restriction enzymes, the mpt51 fragment was ligated into the vector, and the Escherichia coli (E. coli) TOP10 strain were transformed by the recombinant plasmid. Positive clones were identified by colony PCR, restriction enzyme digestion, and DNA sequencing. Results: The mpt51 gene was successfully cloned into pcDNA3.1(+). A 6400 bp band for the pcDNA3.1(+)/mpt51 recombinant plasmid and a 926 bp band for mpt51 were observed by colony PCR, and restriction enzyme digestion on agarose gels. The DNA sequence was 100% homologous with the mpt51 fragment of H37Rv in GenBank. Conclusion: In the current study, the mpt51 gene of MTB was correctly cloned into pcDNA3.1(+). The expression of this recombinant vector can be studied in eukaryotic cells. Moreover, it is possible to determine the efficacy of this vector as a DNA vaccine candidate, and to test its protective function compared to BCG in animal models in future.

Research paper thumbnail of Evaluation of autophagy induction and inhibition in the Huh7.5 cell line through flow cytometry

Medical laboratory sciences, 2017

Background: Autophagy is a physiologic process in which double membrane vesicles engulf damaged p... more Background: Autophagy is a physiologic process in which double membrane vesicles engulf damaged proteins and organelles for delivering them to lysosome in order to degrade and recycle them via lysosomal digestion. Beclin1 is one of the basic proteins involved in the initial step of auto phagosome formation. In the current study, the effects of exogenous Beclin1 and 3-methyladenine (3-MA) to induce and inhibit of autophagy were assessed in Huh7.5 cells as an in vitro models of hepatocellular carcinoma. Materials and Methods: The recombinant pcDNA-Beclin1was transfected into Huh7.5 cells. Also, the cytotoxicity of 3-methyladenine (3-MA) were determined in Huh7.5 cells and the cells were treated with 3-methyladenine (3-MA). The autophagy induction and inhibition was conducted via LC3 staining as a main autophagy marker using flowcytometry. Results: The result of this study suggest that the over expression ofexogenousBeclin1 in Huh7.5 cells elevated the autophagosome formation as shown by intracellular autophagosomal marker LC3-II staining for about 32.32 % and 3-MA decreased it up to2% compared with control cells in which the stained LC3-II was12.08.The IC50 of 3-methyladenine (3-MA) was 7.2 mM in Huh7.5 cells. Conclusion: Recombinant beclin1 may be used as a potential autophagy inducer agent and3methyladenine (3-MA) inhibits autophagy formation in Huh7.5 cell. The staining autophagy formation marker LC3-II with specific antibody is a reliable method to measure autophagy activation via flow cytometry.

Research paper thumbnail of Isolation, cloning and molecular analysis of ag85a and tb10.4 genes from Mycobacterium tuberculosis

Medical laboratory sciences, Oct 16, 2017

Background: Novel tuberculosis (TB) vaccines that aim to boost and/or replace Bacillus Calmette-G... more Background: Novel tuberculosis (TB) vaccines that aim to boost and/or replace Bacillus Calmette-Guerin (BCG) are currently in development. DNA vaccines can stimulate both humoral and cell-mediated immunity in different animal models of TB and is thought to be a promising strategy in the development of new vaccines against TB. The aim of this study was to design and construct a DNA vaccine encoding ag85a and tb10.4 fusion genes of Mycobacterium tuberculosis. Materials and Methods: tb10.4 fragment was amplified by PCR and the product was digested with restriction enzymes. Next, it was cloned into the pcDNA3.1+ plasmid. The ag85a gene and pcDNA3.1+/tb10.4 plasmid were digested by EcoRI and BamHI restriction enzymes. Constructed vector was sequenced. The molecular analysis was done using bioinformatics software. New chimeric vector containing ag85a-tb10.4 genes were purified. Expression of pcDNA3.1+/tb10.4-ag85a plasmid was confirmed in eukaryotic cells. Results: Fragments of 297 bp for tb10.4 and 1017 bp for ag85a were observed in agarose gel electrophoresis. Alignment of ag85a-tb10.4 genome sequence with reference genes in GenBank showed exact identities that indicate correction of all cloning procedures. Transfection of eukaryotic cells with pcDNA3.1+/tb10.4-ag85a vector and existence of tb10.4-ag85a fusion gene were both confirmed with RT-PCR. Conclusion: In this study, tb10.4 and ag85a genes were isolated from Mycobacterium tuberculosis H37Rv strain and cloned into pcDNA3.1+. Also, the capability of constructed vector in producing fusion ag85a-tb10.4 protein was confirmed with RT-PCR. pcDNA3.1+/tb10.4-ag85a vector can be used for further studies in future.

Research paper thumbnail of Comparison of the culture and PCR methods for diagnosis of Mycobacterium tuberculosis in different clinical specimens

Clinical Biochemistry, Sep 1, 2011

Background: Tuberculosis remains a global epidemic, especially in developing countries, including... more Background: Tuberculosis remains a global epidemic, especially in developing countries, including Iran. Rapid diagnosis of active Mycobacterium tuberculosis infection plays a critical role in controlling the spread of tuberculosis. Conventional methods may take up to several weeks or longer to produce results. In addition to multiplicity of steps involved in conventional detection, including isolation, identification and drug susceptibility testing, the slow growth rate of M. tuberculosis is also responsible for this lengthy time. Objectives: The aim of this study was to compare the polymerase chain reaction (PCR) and culture methods for the detection of M. tuberculosis in different clinical specimens. Materials and Methods: This study was performed on different samples (urine, gastric aspirate, bronchoalveolar lavage, pleural fluid, cerebrospinal fluid, ascetic fluid and joint fluid specimens) of tuberculosis suspected patients. M. tuberculosis DNA was extracted directly from different samples using two different protocols. Next, PCR was performed using three sets of specific primers to detect members of Mycobacterium genus, M. tuberculosis complex and non-tuberculosis Mycobacteria. The results were then compared with that of the culture method, which is considered as the gold standard method. Results: The concordance rate between the three sets of primers was calculated and IS6110/buffer PCR method showed good agreement with the LJ culture method (κ = 0.627, P < 0.0001). The sensitivity of IS6110/buffer PCR was 58.33%, with specificity of 77.78%; the positive and negative predictive values were 100% and 78.26%, respectively. Buffer method for DNA extraction was proved to give a higher accuracy to PCR in comparison with the boiling method. Conclusions: PCR method is a valuable, cost-effective and alternative tool for quick diagnosis of active tuberculosis in different clinical specimens.

Research paper thumbnail of Relationship between the prevalence and severity of non‐alcoholic fatty liver disease and coronary artery disease: Findings from a cross‐sectional study of a referral center in northeast Iran

JGH open, Apr 27, 2022

Background and AimNon‐alcoholic fatty liver disease (NAFLD) is becoming increasingly prevalent wo... more Background and AimNon‐alcoholic fatty liver disease (NAFLD) is becoming increasingly prevalent worldwide, and cardiovascular diseases are the most common cause of death in NAFLD patients. The present study aimed to evaluate the possible relationship between the presence and severity of NAFLD and coronary artery disease (CAD).MethodsA cross‐sectional study was conducted on 296 patients (122 men and 174 women, with mean age 54.10 ± 9.33 years) referred to the catheterization laboratory of Imam Reza Hospital affiliated to the Mashhad University of Medical Sciences, Mashhad, Iran, for elective coronary angiography to investigate the presence and severity of CAD. Additionally, all patients underwent abdominal ultrasonography (USG) to detect NAFLD and its severity.ResultsAmong the 296 patients, 187 (63.2%) had CAD and 160 (50.1%) had NAFLD. NAFLD patients had significantly higher prevalence of obesity (odds ratio [OR] = 1.047, 95% confidence interval [CI] = 1.002–1.094), hypertension (OR = 1.909, 95% CI = 1.027–3.55), hyperlipidemia (OR = 3.474, 95% CI = 1.862–6.482), and CAD (OR = 2.009, 95% CI = 1.100–3.669). The percentage of patients with normal vessels was higher in the non‐NAFLD group, followed by the group with mild and severe NAFLD (P < 0.001). However, single‐ and multi‐vessel disease incidences among the non‐NAFLD, mild, and severe NAFLD groups were 36.1, 43.1, and 63.7%, respectively. Interestingly, the percentage of patients with two‐vessel stenosis was significantly higher in severe NAFLD patients than mild and non‐NAFLD patients (P < 0.001).ConclusionThe prevalence and severity of NAFLD were independently associated with CAD. Mild NAFLD was primarily observed among patients with normal and non‐obstructive coronary artery patients, while severe NAFLD was more frequent in extensive CAD patients with multi‐vessel disease.

Research paper thumbnail of A case of recurrent meningitis caused by drug resistant Enterococcus fecalis in a patient with ventriculoperitoneal shunt

KAUMS Journal, Jul 15, 2007

ABSTRACT Background: Iatrogenic meningitis is an important complication in neurosurgery. This inf... more ABSTRACT Background: Iatrogenic meningitis is an important complication in neurosurgery. This infection is not common. This paper presents the first report of recurrent shunt meningitis caused by drug resistant Enterococcus fecalis in Iran. Case report: We report a case with recurrent shunt meningitis caused by Enteroccocus fecalis in a 9-year old girl with ventriculoperitoneal shunt which did not recover by antibiotic therapy until the removal of the infected shunt. Discussion and Conclusion: Patients with hydrocephalus who are treated with ventriculoperitoneal shunts are susceptible to bacterial infections. The rate of this nosocomial infection varies from 2% to 22%. It is suggested to pay more attention to symptoms of CNS infections among patients with cerebrospinal fluid drainage. Keywords: Recurrent meningitis, Ventriculoperitoneal shunt, Enteroccocus fecalis, Drug resistant,

Research paper thumbnail of Designing a label‐free electrochemical aptasensor based on polypyrrole‐<scp>l</scp>‐cysteine‐reduced graphene oxide nanocomposite for detection of 25‐hydroxyvitamin D<sub>3</sub>

Biotechnology and Applied Biochemistry, Jun 27, 2023

Research paper thumbnail of Synthesis of γ-Fe2O3 Nanoparticles Capped with Oleic Acid and their Magnetic Characterization

Iranian Journal of Science and Technology Transaction A-science, Feb 22, 2017

In recent years, superparamagnetic iron oxide nanoparticles have attracted a great attention due ... more In recent years, superparamagnetic iron oxide nanoparticles have attracted a great attention due to their various biomedical applications, such as magnetic resonance imaging, targeted drug delivery, and hyperthermia. In this article, c-Fe 2 O 3 magnetic nanoparticles (Maghemite) were prepared in oleic acid media by co-precipitation method. The oleic acid, a monounsaturated fatty acid was used as the capping and stabilizing agent during the synthesis of the magnetic nanoparticles. Characterization of obtained nanoparticles were performed using powder X-ray diffraction (PXRD), field emission scanning electron microscopy (FESEM), Fourier transform infrared spectra (FTIR), and vibrating sample magnetometer (VSM). The crystallite size of c-Fe 2 O 3 nanoparticles was achieved in the range between 16.2 and 26.8 nm. The FESEM demonstrated the regular spheres of c-Fe 2 O 3 nanoparticles. The obtained nanoparticles were coated with oleic acid indicating by FTIR analysis. The resulted oleic acid-coated nanoparticles were shown superparamagnetic properties (*52 emu/g). This suggested method is simple and rapid to fabricate superparamagnetic nanoparticles which make them appropriate candidates for theranostic application in future studies.

Research paper thumbnail of Artemisia Absinthium Extract Attenuates the Quinolinic Acid-Induced Cell Injury in OLN-93 Cells

Current Drug Discovery Technologies

Objective: Increased quinolinic acid (QA) accumulation has been found in many neurodegenerative d... more Objective: Increased quinolinic acid (QA) accumulation has been found in many neurodegenerative diseases. Artemisia absinthium (A. absinthium) has been reported to have neuroprotective and antioxidant activities. This study was designed to evaluate the effect of A. absinthium in QAinduced neurotoxicity in OLN-93 Cells. Methods: OLN-93 cells were cultured in a DMEM medium containing 10% (v/v) fetal bovine serum, 100 units/ml penicillin, and 100 μg/ml streptomycin. The cells were pretreated with concentrations of A. absinthium extract for two h and then exposed to QA for 24 h. After 24 h cell viability, the level of malondialdehyde (MDA), reactive oxygen species (ROS), and apoptotic cells were quantitated in OLN-93 Cells. Results: Pretreatment with A. absinthium extract prevented the loss of cell viability in OLN-93 cells. ROS generation, lipid peroxidation, and apoptosis in QA -injured OLN-93 cells were reduced following A. absinthium extract pretreatment. Conclusion: A. absinthium e...

Research paper thumbnail of Relevance of aptamers as targeting ligands for anticancer therapies

Research paper thumbnail of Seroprevalence of Herpes Simplex Viruses Types 1 and 2 in a Population, Age 15-35 Years, of Mashhad City

Iranian biomedical journal, Mar 1, 2023

Background: Considering the high prevalence and clinical importance of herpes simplex virus (HSV)... more Background: Considering the high prevalence and clinical importance of herpes simplex virus (HSV) infection worldwide, we aimed to evaluate the seroprevalence of HSV-1 and HSV-2 in a population aged between 15 and 35 years in Mashhad, Iran. Methods: This cross-sectional study was conducted on 916 cases composed of 288 (31.4%) men and 628 (68.6%) women. Using ELISA method, the presence of IgM and IgG antibodies against HSV-1 and HSV-2 was assessed. Results: Among the population studied, 681 (74.3%) cases were positive for anti-HSV antibodies, while 235 (25.7%) cases were negative. Moreover, no IgMs were found and all positive subjects had IgG antibodies. Age (p < 0.001), occupation (p < 0.001), education (p = 0.006), smoking (p = 0.029), and BMI (p = 0.004) demonstrated a significant association with HSV-1 and HSV-2 infection. Conclusion: Our study indicates a high seroprevalence of HSV infection; however, there was no cases positive for IgM antibodies, suggesting the high prevalence of latent infection.

Research paper thumbnail of Herpes Simplex Infection in Cardiovascular Disease (CVD) Patients: A Seroepidemiological Study

Journal of Kermanshah University of Medical Sciences, Mar 14, 2023

Background: Herpes simplex viruses have been implicated as a cause of cardiovascular disease (CVD... more Background: Herpes simplex viruses have been implicated as a cause of cardiovascular disease (CVD) due to their widespread distribution and ability to infect human vascular endothelial cells. Objectives: The present study aimed to evaluate the link between Herpes simplex infections and cardiovascular disease. Methods: This case-control study involved 236 patients aged 35-65, 118 with known cardiovascular disease and 118 controls. Patients' cardiovascular disease evaluation was based on data from questionnaires, a specialist's physical examinations, and paraclinical tests. Herpes simplex viruses (HSV)-antibodies (IgM, IgG) were determined using ELISA in serum samples, and the biochemical blood indicators were analyzed to examine their relationship to the level of HSV antibodies. All the data were analyzed using SPSS software version 20. Results: IgM antibodies against herpes simplex were negative among all 236 patients. The positivity rate of IgG antibodies against herpes simplex was 58.8 and 51.2% in the case and control groups, respectively with no significant difference (P = 0.253). Patients with cardiovascular disease had a 0.587-fold higher positive rate of IgG antibodies than patients in the control group (odds ratio (OR) = 0.587). The mean age of patients with positive IgG antibodies was significantly lower than patients with negative IgG antibodies (P = 0.012). IgG positivity among men and women (P = 0.670) and different job statuses (P = 0.866) were not significantly different. Conclusions: The positivity rate of IgG antibodies against herpes simplex was not significantly different among the study groups. Although patients with positive IgG antibodies' mean age was lower than those with negative IgG antibodies, the gender and job status were not different.

Research paper thumbnail of A new DNA vaccine expressing HspX-PPE44-EsxV fusion antigens of Mycobacterium tuberculosis induced strong immune responses

PubMed, Jul 1, 2020

Objectives: Infection with tuberculosis (TB) is regarded as a major health issue. Due to the emer... more Objectives: Infection with tuberculosis (TB) is regarded as a major health issue. Due to the emergence of antibiotic resistance during TB treatment, prevention via vaccination is one of the most effective ways of controlling the infection. DNA vaccines are developed at a greater pace due to their ability in generating a long-lasting immune response, higher safety compared to the live vaccines, and relatively lower cost of production. In the present study, we evaluated a new DNA vaccine encoding the fusion HspX-PPE44-EsxV antigens, separately, and in combination with Bacillus Calmette-Guérin (BCG) administration, in a prime-boost method in mice. Materials and methods: A novel DNA vaccine encoding HspX-PPE44-EsxV fusion antigen of Mycobacterium tuberculosis was constructed, and RT-PCR and Western blot analysis were performed to verify the expression of the antigen. Female BALB/c mice were divided into five groups (PBS, BCG, pcDNA3.1 (+) vector, pDNA/HspX-PPE44-EsxV vaccine, and the BCG-prime boost groups). In order to evaluate the immunogenicity of the recombinant vector, BALB/c mice were injected with 100 μg of pDNA at 2-week intervals. Then, cytokine assay was conducted using eBioscience ELISA kits (Ebioscience, AUT) according to manufacturers' instructions to evaluate the concentrations of IL-4, IL-12, TGF-β, and IFN-γ. Results: The concentrations of INF-γ, IL-12, and TGF-beta were significantly increased compared to the control groups (P<0.001). INF-γ and IL-12 production were increased significantly in pDNA/HspX-PPE44-EsxV+BCG group compared to pDNA/HspX-PPE44-EsxV group (P<0.001). Conclusion: This study showed that the present DNA vaccine could induce a high level of specific cytokines in mice. It was also shown that using this DNA vaccine in a BCG prime-boost protocol can produce significant amounts of IFN-γ, IL-12, and TGF-β.

Research paper thumbnail of Designing and construction a DNA vaccine encoding the fusion fragment of cfp10 and ag85a immunodominant genes of Mycobacterium tuberculosis

Medical laboratory sciences, May 21, 2017

Background: Pathogenic mycobacteria are one of major causes of human morbidity and mortality. Myc... more Background: Pathogenic mycobacteria are one of major causes of human morbidity and mortality. Mycobacterium tuberculosis (M. tuberculosis) is an etiological agent of human tuberculosis. Designing new vaccines including DNA vaccines may be considered as new approaches for preventing of TB. Materials and Methods: M. tuberculosis H37Rv was grown on Lowenstein Jensen medium for 4 weeks at 37ºC and then DNA was extracted. The cfp10 gene was amplified by PCR. After digesting the PCR product and the plasmid, cfp10 fragment was ligated into the vector using T4 DNA ligase. Then, Ag85A was subcloned into pcDNA/cfp10. Escherichia coli strain JM109 bacteria were transformed by the desired construct. Clone confirmations were performed by colony PCR, restriction enzyme digestion and DNA sequencing. Recombinant vector was transfected into HeLa cells and total RNA was extracted, then cDNA was synthesized using oligo-dT. Finally PCR was performed by cfp10 primers. Results: The cfp10 was amplified by PCR method and the PCR products were visualized by agarose gel electrophoresis. The cfp10 fragments showed 303 bp in length. The cfp10 cloned into pcDNA. Then, Ag85Awas ligated into pcDNA/cfp10 after digestion correctly. Colony-PCR and restriction enzyme digestion and sequencing confirmed the cloning the fusion Ag85A/cfp10 fragment. Finally, after cDNA synthesis, expression of vector was confirmed in eukaryotic system. Conclusion: Cloning of Ag85A/cfp10 genes of M. tuberculosis were performed correctly. It can use as a DNA vaccine for investigation the immune responses in animal models in future studies.

Research paper thumbnail of CTLA-4 Gene Haplotypes and the Risk of Chronic Hepatitis C Infection; a Case Control Study

PubMed, Oct 1, 2017

Background: The prevalence of hepatitis C virus (HCV) infection is increasing worldwide. Cytotoxi... more Background: The prevalence of hepatitis C virus (HCV) infection is increasing worldwide. Cytotoxic Tlymphocyte-associated protein 4 (CTLA-4) may play a role in the intensity of the disease. The aim of this study was to evaluate the association between genetic variants of the CTLA-4 and HCV infection. Methods: Restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) was performed as the genotyping assay at four different positions (+49 A>G, -318 C>T, -1722 T>C, and - 1661 A>G). Haplotypes were analyzed using PHASE software. Sixty-five HCV patients and 65 healthy individuals as controls who were referred to the hepatitis clinic in Mashhad, Iran, were recruited. Genomic DNA was extracted from whole blood of participants. Results: In a dominant analysis model of the -1661 position (GG vs. AA+AG), the AA genotype was more common in controls than in patients (adjusted P = 0.0003; OR = 0.15, 95% CI = 0.051 -0.42). The GCAT haplotype was also more prevalent in controls than in patients (adjusted P = 0.01; OR = 0.40, 95% CI = 0.20-0.81). Furthermore, the ACGT/ACGT diplotype was more common in controls than in patients (P = 0.0037; OR = 0.15, 95% CI = 0.04-0.54). In addition, the ACGT/ACAT diplotype was more frequent in patients than controls (adjusted P =0.003; OR = 2.48, 95% CI = 1.37- 4.50). Conclusion: Our results indicated that polymorphisms in CTLA-4 and certain haplotypes may affect the risk of HCV infection in our population, although a larger sample size may be required to confirm this association.

Research paper thumbnail of The Antiviral Effects of Curcumin Nanomicelles on the Attachment and Entry of Hepatitis C Virus

Iranian Journal of Virology, Jun 15, 2017

Background and Aims: Hepatitis C virus (HCV) is a member of the Flaviviridae family, which causes... more Background and Aims: Hepatitis C virus (HCV) is a member of the Flaviviridae family, which causes approximately 500,000 deaths annually. HCV infection treatment is often associated with significant adverse effects. Curcumin is an active ingredient of turmeric which has therapeutic anti-inflammatory effects in many diseases including infectious ones. Although curcumin is not soluble in water, if it is synthesized in the form of nanomicelles, it will be water soluble and can be absorbed in the gastrointestinal tract (GI). In this study, the antiviral effects of curcumin nanomicelles were investigated on the attachment and entry of HCV particles. Materials and Methods: The cytotoxicity of curcumin nanomicelles was determined in Huh7.5 cells and their antiviral effects on the attachment and entry of HCV was investigated in a cell culture system. Results: Curcumin nanomicelles could decrease the viral load in the cell culture supernatants compared to virus control. Conclusions: According to the results of this research, we determined the antiviral effects of curcumin nanomicelles in the later stages of HCV replication.

Research paper thumbnail of Evaluation of Immune Responses to a DNA Vaccine Encoding Ag85a-Cfp10 Antigen of Mycobacterium tuberculosis in an Animal Model

Jundishapur Journal of Microbiology, Dec 16, 2018

Background: Many studies indicate that the Bacillus Calmette-Guérin (BCG) vaccine has low protect... more Background: Many studies indicate that the Bacillus Calmette-Guérin (BCG) vaccine has low protective efficacy, especially in endemic areas. There are several factors in this assessment, such as genetic diversity of BCG strains, pre-exposure to environmental mycobacteria, and variations in host immune responses. Currently, more than 200 new vaccine candidates have been proposed, such as recombinant BCG, DNA, and subunit vaccines. However, none of them are superior to BCG. Nevertheless, several approaches are considered to reduce the cases of tuberculosis infection. Objectives: The aim of the present study was to evaluate the capability of the Ag85a-cfp10 fusion protein as a new chimeric protein in stimulating immune responses. Methods: A DNA vaccine encoding Ag85a-cfp10 fusion protein was constructed in the previous study. The expression of Ag85a-cfp10 fusion protein in host cells was confirmed by the RT-PCR method. Six pathogen-free female mice were injected intramuscularly at a total concentration of 100 µg/mL three times at two-week intervals. The BCG and the control groups received BCG and PBS vaccines, respectively. One month after the final immunization, mice were killed and their splenocytes were cultured in RPMI medium supplemented with 1% antibiotics and 10% serum. Four cytokines including IL-4, IL-12, TGF-β, and IFN-γ were measured in the culture supernatant using the ELISA test. Results: RT-PCR analysis showed that Ag85a-cfp10 recombinant vector is able to replicate in eukaryotic cells and produce mRNA. The vaccinated groups were compared to the control group, showing induction of high levels of cytokine production. Conclusions: Some reports depicted that DNA vaccines are able to induce humoral and cellular immune responses both in animal models and humans. Therefore, in the current study, the immune response was induced in mice, which were inoculated with recombinant expression plasmid, pcDNA3.1 (+)-Ag85a-cfp10. We showed that this recombinant vector can stimulate mycobacterial specific modulating cytokines. Nonetheless, analysis appeared that this vaccine is unable to stimulate cell mediated immunity, however, still further studies are needed in future.