Publications by authors named "Amitis Ramezani"

93 Publications

SARS-CoV-2 presented moderately during two episodes of the infection with lack of antibody responses.

Virus Res 2021 Apr 6:198421. Epub 2021 Apr 6.

Clinical Research Department, Pasteur Institute of Iran, Tehran, Iran. Electronic address:

The world has gone through the critical phase of SARS-CoV-2 crisis caused by the new variants of the virus. The globally concerted effort to characterize viral genomic mutations across different clades has revealed several changes in the coding and also non-coding regions which might lead to a violent presentation or re-infection occurrence. Here, we studied a COVID-19 subject who represented the symptoms following the full recovery of the first infection. COVID-19 specific IgM and IgG were evaluated in both steps. The viral samples from oropharyngeal/nasopharyngeal were subjected to RT-PCR and full sequencing was done in both incidences. The sequencing data was fully investigated with the reference sequence of SARS-CoV-2 and the changes were detected. The obtained data is in favor of re-infection with 128 days of interval. SARS-CoV-2 presented more severely in the second episode of the disease and the specific antibodies against COVID-19 were not detectable. Both infections were caused by the same clade 20 G, however, the mutation rates were higher in the second incidence including 10 nucleotide substitutions which had rarely been reported before. In the present study, the nucleotide mutations in various regions of the viral genome have been presented. The re-infection could have significant effect on clinical implications as well as vaccination.
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http://dx.doi.org/10.1016/j.virusres.2021.198421DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8022517PMC
April 2021

Evaluation of transduced dendritic cells expressing HIV-1 p24-Nef antigens in HIV-specific cytotoxic T cells induction as a therapeutic candidate vaccine.

Virus Res 2021 Mar 25:198403. Epub 2021 Mar 25.

Clinical Research Department, Pasteur Institute of Iran, Tehran, Iran. Electronic address:

Various approaches have been investigated to prevent or eliminate HIV-1 since 1981. However, the virus has been affecting human population worldwide with no effective vaccine yet. The conserved regions among the viral genes are suitable targets in mutable viruses to induce the immune responses via an effective delivery platform. In this study, we aimed at evaluation of p24 and nef in two forms of full and truncated genes as two fusion antigenic forms according to our previous bioinformatics analysis. The designed antigens were then transferred through ex vivo generated dendritic cells and also proteins in BALB/c to assess and compare immunogenicity. p24 and Nef amino acid sequences were aligned, then, the most conserved regions were selected and two fusion forms as the truncated (p24:80-231aa-Nef:120-150aa) and the full from (p24-Nef) were cloned and expressed in prokaryotic and eukaryotic systems. Lentiviral vectors were applied to generate recombinant virions harboring the genes of interest to transduce generated murine dendritic cells. BALB/c mice received the recombinant DCs or recombinant proteins according to the defined schedule. IgG development was assessed to determine humoral immune activity and cellular immune responses were evaluated by IL-5 and IFN-y induction. Granzyme B secretion was also investigated to determine CTL activity in different immunized groups. The data showed high induction of cellular immune responses in dendritic cell immunization specifically in immunized mice with the truncated form of the p24 and Nef by high secretion of IFN-y and strong CTL activity. Moreover, protein/ DC prime-boost formulation led to stronger Th1 pathway and strong CTL activation in comparison with other formulations. The generated recombinant dendritic cells expressing p24-Nef induced humoral and cellular immunity in a Th1 pathway specifically with the in silico predicted truncated antigen which could be of high value as a dendritic cell therapeutic vaccine candidate against HIV-1.
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http://dx.doi.org/10.1016/j.virusres.2021.198403DOI Listing
March 2021

Prolonged Viral Shedding and Antibody Persistence in Patients with COVID-19.

Microbes Infect 2021 Mar 16:104810. Epub 2021 Mar 16.

Clinical Research Department, Pasteur Institute of Iran, Tehran, Iran. Electronic address:

SARS-CoV-2 as a new global threat has affected global population for one year. Despite the great effort to eradicate this infection, there are still some challenges including different viral presentation, temporal immunity in infected individuals and variable data of viral shedding. We studied 255 COVID-19 suspected individuals to assess the viral shedding duration and also the antibody development against SARS-CoV-2 among the cases. Real Time RT-PCR assay was applied to determine the virus presence and SARS-CoV-2 antibodies were evaluated using SARS-CoV-2 IgM and IgG kits. 113 patients were confirmed for COVID-19 infection. The patients were followed until negative PCR achieved. The median viral shedding among studied population was obtained 34.16 (±17.65) days which was not significantly associated with age, sex and underlying diseases. Shiver and body pain were found in prolonged form of the infection and also patients who had gastrointestinal problems experienced longer viral shedding. Moreover, IgG was present in 84% of patients after 150 days. According to this data, the median viral shedding prolongation was 34.16 days which indicates that 14 days isolation might not be enough for population. In addition, IgG profiling indicated that it is persistent in a majority of patients for nearly 6 months which has brought some hopes in vaccine efficacy and application.
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http://dx.doi.org/10.1016/j.micinf.2021.104810DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7963517PMC
March 2021

Clinical characteristics of SARS-CoV-2 by re-infection vs. reactivation: a case series from Iran.

Eur J Clin Microbiol Infect Dis 2021 Mar 18. Epub 2021 Mar 18.

Clinical Research Department, Pasteur Institute of Iran, Tehran, 1316943551, Iran.

COVID-19 immunity in infected individuals may not be persistent. The specific response wanes in patients who have recovered from this infection. Nevertheless, it has not been fully understood whether true re-infection occurs or the viral reactivation. In this study, we investigated three COVID-19 patients who represented the symptoms after recovery. Chest CT scan was applied to assess the patients along with the viral samples from oropharyngeal/nasopharyngeal which were subjected to RT-PCR. The viral genome sequencing was applied where possible to distinguish possible re-infection or latent reactivation. Moreover, COVID-19-specific antibodies available data were evaluated in each incidence. The second episode of SARS-CoV-2 infection was different among the investigated subjects who experienced an interval between positive PCR tests ranged between 63 and 156 days. The disease presentation was less or more severe in the second infection. All cases were found IgG positive in the re-infection phase. The sequencing of SARS-CoV-2 sample obtained from two cases revealed a D614G mutation of S gene from the second isolated sample strengthens the case for the re-infection. The possibility of re-infection and reactivation could have significant effect on clinical implications and also vaccination. Our data supports clear warning of SARS-CoV-2 continuous circulation potency among the populations in spite of herd immunity either with natural infection or vaccination. This issue is critical in term of the patients, clinical investigate, and viral transmission.
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http://dx.doi.org/10.1007/s10096-021-04221-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7972329PMC
March 2021

A review study about the effect of chitosan nanocarrier on improving the efficacy of amphotericin B in the treatment of leishmania from 2010 to 2020.

Curr Drug Deliv 2021 Mar 16. Epub 2021 Mar 16.

Department of Clinical Research, Pasteur Institute of Iran, Tehran. Iran.

In the present review study, the published articles from 2010-2020 that evaluated the effect of chitosan nanocarrier on the amphotericin B (AmB) efficacy in the treatment of leishmaniasis, have been considered. Leishmania is a parasitic tropical disease in the world and is treated with AmB as one of the main therapeutic agent. However, clinical application of AmB is limited due to its toxicity and insolubility issues. Using nanoparticles and in particular chitosan nanocarrier seems a promising approach to overcome these problems. Therefore, various doses of AmB have been loaded in chitosan nanoparticles in different studies and the results of these studies demonstrated that by increasing the drug loading efficiency and decreasing the toxicity, the potency of the nanoformulation to inhibit and killing the parasite is increased. In this regard, the results of a study performed in 2018, demonstrated that chitosan nanoparticles with the higher dose of drug loading were the most effective formulation to inhibit and kill the parasite. Thus, chitosan nanocarrier can consider as an appropriate candidate in the future to inhibit and kill the leishmania parasite without causing side effects.
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http://dx.doi.org/10.2174/1567201818666210316111941DOI Listing
March 2021

HIV-1 p24-nef DNA Vaccine plus Protein Boost Expands T-Cell Responses in BALB/c.

Curr Drug Deliv 2020 Dec 31. Epub 2020 Dec 31.

Clinical Research Department, Pasteur Institute of Iran, Tehran. Iran.

Background: There have been massive efforts on vaccine development against HIV-1 since its discovery. Various approaches have been taken to attention including rational vaccine design, optimized delivery systems and heterologous regimen to eradicate the virus. DNA vaccines fundamentally induce host immune responses by genetically engineered plasmids encoding antigens and expressed in vivo without need of specific delivery system. Therefore, a longterm of endogenous antigen expression could be possible.

Objective: In this study, we aimed at evaluation and comparison of DNA and protein vaccine based on two forms of full and truncated HIV-1 p24-nef antigens by in silico design in BLALB/c.

Methods: The recombinant pcDNA3.1 harboring two sets of HIV-1 p24 and nef genes in truncated and full forms were generated and applied to immunize BALB/c along with the corresponding proteins via three different DNA/DNA, DNA/protein and protein/protein regimens.

Results: The results showed that the applied regimens could elicit strong immune responses in comparison with controls and the prim-boost DNA/protein regimen reached the highest immune induction (p < 0.05). Moreover, prime-boost approach was assessed more successful in a qualitatively broad Th1 response induction. The truncated form of the antigens, p24(80- 231 aa)-AAY- Nef (120-150), was evaluated more immunogenic in agreement with the in silico investigation.

Conclusion: The truncated form of p24-Nef was evaluated highly immunogenic specially when applied in prim-boost DNA/Protein regimen and could be investigated in other delivery systems and a proper animal model to achieve a therapeutic vaccine candidate against HIV-1.
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http://dx.doi.org/10.2174/1567201818666210101113601DOI Listing
December 2020

SARS-CoV‑2, a virus with many faces: a series of cases with prolonged persistence of COVID-19 symptoms.

Wien Med Wochenschr 2021 Feb 14;171(1-2):3-6. Epub 2020 Dec 14.

Clinical Research Department, Pasteur Institute of Iran, No:69, Pasteur Ave, 1316943551, Tehran, Iran.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), as the causative agent of the ongoing pandemic, has spread into more than 200 countries to date. The disease which is caused by the virus is termed COVID-19. In most cases, it presents at first like common flu with cough and other respiratory symptoms. Nevertheless, other symptoms have been reported, such as a feeling of extreme fatigue, gastrointestinal symptoms, or acute onset of olfactory and gustatory dysfunction. Here we report a series of 10 cases (1 male, 9 females) observed between February and April 2020, with an undulating appearance and disappearance of symptoms. Weeks passed before the diagnosis was established. Symptoms resolved rapidly after treatment with hydroxychloroquine. It seems that the course of COVID-19 can be mild or moderate but with a long persistence of symptoms, and may therefore remain obscure. This may cause a public health issue because of the long infectivity of these patients.
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http://dx.doi.org/10.1007/s10354-020-00793-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7734614PMC
February 2021

Importance of evaluating occult HBV risk factors in a low endemic area for HBV infection.

Wien Med Wochenschr 2021 02 28;171(1-2):16-17. Epub 2020 Sep 28.

Clinical Research Department, Pasteur Institute of Iran, No: 69, Pasteur Ave, 1316943551, Tehran, Iran.

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http://dx.doi.org/10.1007/s10354-020-00784-9DOI Listing
February 2021

No evidence of occult HBV infection in population born after mass vaccination.

Wien Med Wochenschr 2020 Jun 9;170(9-10):218-223. Epub 2020 Apr 9.

Clinical Research Dept., Pasteur Institute of Iran, Pasteur Ave., 13164, Tehran, Iran.

Despite access to efficient hepatitis B virus (HBV) vaccine and universal immunization schedules, HBV infection remains a global health concern. HBV infection has decreased by this program. Nevertheless, breakthrough infections occur due to generation of occult HBV infection (OBI) and surface gene mutants in the immunized population. We aimed to determine the presence of OBI in a population born after initiation of nationwide HBV vaccination in Tehran, Iran. A HBV mass vaccination schedule was launched in Iran in 1993. For this study, we enrolled 1120 cases younger than 24 years. ELISA was applied to evaluate the presence of HBsAg, anti-HBs and anti-HBc. HBV-DNA presence was determined in all HBsAg-negative cases using nested polymerase chain reaction. The prevalence of HBsAg, anti-HBc and anti-HBs was 0.1, 0.54 and 39.9% respectively. Out of 6 anti-HBc-positive individuals, 4 cases also had anti-HBs. One case revealed HBsAg co-existence and the other one showed isolated anti-HBc. HBV-DNA was not detected in HBsAg-negative specimens. A very low prevalence of HBsAg and isolated anti-HBc was observed and no occult HBV infection was detected. It seems that evasion mutants are not a potential threat for HBV universal immunization efficacy in the vaccinated population.
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http://dx.doi.org/10.1007/s10354-020-00748-zDOI Listing
June 2020

Production of Recombinant HIV-1 p24-Nef Protein in Two Forms as Potential Candidate Vaccines in Three Vehicles.

Curr Drug Deliv 2020 ;17(5):387-395

Department of Clinical Research, Pasteur Institute of Iran, Tehran, Iran.

Background: Different approaches have been investigated to develop a preventive or therapeutic vaccine, although none of them has been fully practical. Therapeutic vaccines against HIV-1 have been studied with the aim of eliminating the virus from reservoir cells with or without HAART (Highly Active Antiretroviral Therapy). Fusion proteins with the most immunogenic features among conserved regions can facilitate this achievement in such a variable virus. To achieve the most immunogenic and also conserved regions, bioinformatics tools are widely used to predict antigens' features before applying them.

Objective: This study aimed at the in vitro evaluation of p24 -Nef fusion protein based on the previous in silico design to achieve a potential therapeutic subunit vaccine against HIV-1.

Methods: The truncated form of p24-Nef using AAY flexible linker and the full protein were expressed and evaluated in the prokaryotic system and confirmed by western blotting. We also used pcDNA3.1 to transfect Lenti-X 293T cells. Moreover, lentiviral vectors were applied to produce recombinant virions harboring the genes of interest and cell transduction.

Results: Both fusion proteins in a truncated and a full form were expressed and confirmed by Anti Nef polyclonal antibody in western blotting. Recombinant virions were generated and transduced Lenti-X 293T cells confirming by immunofluorescence microscope and p24 ELISA assay kit. Transduced cells were analyzed by SDS-PAGE and western blotting, which resulted in approved protein expression.

Conclusion: Fusion protein of p24 and Nef is well expressed in eukaryotic cell lines according to its pre-evaluated features by bioinformatics tools.
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http://dx.doi.org/10.2174/1567201817666200317121728DOI Listing
January 2020

Comparative analysis between four model nanoformulations of amphotericin B-chitosan, amphotericin B-dendrimer, betulinic acid-chitosan and betulinic acid-dendrimer for treatment of : real-time PCR assay plus.

Int J Nanomedicine 2019 24;14:7593-7607. Epub 2019 Sep 24.

Department of Clinical Research, Pasteur Institute of Iran, Tehran, Iran.

Background: Amphotericin B (Amp) and Betulinic acid (BA) as antileishmanial agents have negligible water solubility and high toxicity. To solve these problems, for the first time, chitosan nanoparticles and Anionic Linear Globular Dendrimer (D) were synthesized for the treatment of (.

Method: Chitosan and dendrimer nanoparticles were synthesized, and Amp and BA were loaded into the nanoparticles. The particles were then characterized using various methods and their efficacy was evaluated in vitro and in vivo environments (parasite burden was confirmed using pathological studies and real-time PCR methods).

Result: The results of docking showed that Amp and BA can be loaded into chitosan and dendrimer nanoparticles. The results of physically drug loading efficiency for AK (Amphotericin B-chitosan), BK (Betulinic acid-chitosan), AD (Amphotericin B-Dendrimer) and BD (Betulinic acid- Dendrimer) were 90, 93, 84 and 96 percent, respectively. The characterization results indicated that the drugs were loaded into nanoparticles physically. Moreover, the increased solubility rate for AD=478, BD=790, AK=80 and BK=300 folds. Furthermore, the results of the drug delivery system showed the slow controlled drug release pattern with cellular uptake of more than 90%. The treatment results showed a 100 percent decrease of toxicity for the all nanodrugs was observed in vivo and in vitro environments. Moreover, AK10 and BK20 mg/kg reduced parasite burden by 83 percent (<0.001), while AD50 and BD40 mg/kg reduced it to a lesser extent compared to glucantime.

Conclusion: All the synthesized nanodrugs were completely succeeded by 100% to recovery the induced pathological effects in the infected footpad. Also, the results of present study were confirmed with real-time PCR and the results showed that AK and BK were succeeded in a large extent to the treatment of infection (<0.001), therefore AK and BK could be considered as proper alternatives of choices drugs.
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http://dx.doi.org/10.2147/IJN.S220410DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6831986PMC
February 2020

Prevalence of Antibodies and DNA in Iranian HIV Patients.

Iran J Pathol 2019 27;14(1):68-75. Epub 2018 Dec 27.

Infectious Diseases Specialist, Dept. of Clinical Research, Pasteur Institute of Iran, Tehran, Iran.

Background & Objective: infection has public health importance and can lead to serious diseases in immunosuppressed patients, such as HIV cases. Appropriate control of infection in HIV patients requires information about the prevalence of antibodies and DNA in different population. In this study, we aimed to determine the prevalence of antibodies and DNA in HIV patients in Tehran, Iran.

Methods: A total of 149 HIV patients from the Iranian Research Center for HIV/AIDS, Tehran, Iran were enrolled in the study. Anti-Toxoplasma IgG and IgM were detected by ELISA and DNA was evaluated by PCR and quantita- tive real-time PCR. IgG positive samples were also assessed for their avidity.

Results: Anti-Toxoplasma IgG and IgM were positive in 46.3% and 2.7% of cases respectively. 92.7% of our patients showed past infection and 4.3% revealed recently acquired toxoplasmosis based on their IgG avidity test. T. gondii DNA was not detected by PCR but real-time PCR results showed DNA in 4.7% of total patients and 13.1% of the IgG seropositive cases.

Conclusion: Our findings indicated that latent toxoplasmosis was relatively prevalent in our study population, but new infection had low prevalence. Almost half of our patients were IgG negative and at risk of acquiring toxoplasma infection. Low copy numbers of DNA were detected in 4.7% of the cases without any clinical manifestation. Therefore, detection and monitoring of anti-Toxoplasma antibodies and DNA in HIV patients is substantial to estimate the risk of reactivation and new infection.
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http://dx.doi.org/10.30699/IJP.14.1.68DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6708569PMC
December 2018

Harmonized Autophagy Versus Full-Fledged Hepatitis B Virus: Victorious or Defeated.

Viral Immunol 2019 10 4;32(8):322-334. Epub 2019 Sep 4.

Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran.

Autophagy is a finely tuned process in the regulation of innate immunity to avoid excessive inflammatory responses and inflammasome signaling. In contrast, the results of recent studies have shown that autophagy may disease-dependently contribute to the pathogenesis of liver diseases, such as fibrosis, cirrhosis, and hepatocellular carcinoma (HCC) during hepatitis B virus (HBV) infection. HBV has learned to subvert the cell's autophagic machinery to promote its replication. Given the great impact of the autophagy mechanism on the HBV infection and HCC, recognizing these factors may be offered new hope for human intervention and treatment of chronic HBV. This review focuses on recent findings viewing the dual role of autophagy plays in the pathogenesis of HBV infected hepatocytes.
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http://dx.doi.org/10.1089/vim.2019.0042DOI Listing
October 2019

Epidemiologic and clinical findings of children with acquired immunodeficiency syndrome in four provinces of Iran.

Wien Med Wochenschr 2020 Jun 13;170(9-10):212-217. Epub 2019 Aug 13.

Pediatric Infectious Disease Research Center, Tehran University of Medical Science, Tehran, Iran.

Introduction: The human immunodeficiency virus (HIV) is still a remarkable challenge throughout the world, especially in developing countries. Despite the fact that HIV in children is becoming one of the most challenging diseases, it seems that pediatric AIDS in Iran is an unknown disease and there is a lack of studies about it. The aim of this study was to describe the characteristics of HIV-positive children who referred to the hospitals of Tehran, Kermanshah, Kurdistan, and Qom provinces of Iran.

Materials And Methods: This study was a retrospective investigation of medical records among 61 children with a diagnosis of HIV who were admitted to the children's hospitals in the four provinces of Iran during a 3-year period (2013-2016).

Results: The frequency of HIV in the center of Iran (Tehran and Qom provinces) was higher (N = 37, 61%). Most of the infected patients were between 5 and 15 years old, 52% were male, and 93% had a history of HIV in their family. Median age at diagnosis of HIV was 2 years. Most of the hospitalized patients were discharged and only two patients (3%) died due to HIV infection. The vast majority of patients (93%) were infected through maternal transmission and a low percentage (29%) were diagnosed before 1 year of age. All of them were in the third stage of the disease. All patients had a positive HIV PCR test. HIV EIA was positive in the majority of cases (98%). A history of addiction in the family was demonstrated in 22 cases (36%). Weight loss (N = 51, 84%), prolonged fever (N = 50, 81%), and respiratory infection (N = 26, 47%) were the most common symptoms.

Conclusion: This study demonstrated a high frequency of pediatric HIV among children aged 5-15 years in four provinces of Iran. Novel strategies to prevent and eliminate mother-to-child transmission of HIV, diagnostic facilities, and treatment of infected mothers during pregnancy in our country are highly recommended.
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http://dx.doi.org/10.1007/s10354-019-0703-1DOI Listing
June 2020

Updated Studies on the Development of HIV Therapeutic Vaccine.

Curr HIV Res 2019 ;17(2):75-84

Hepatitis, AIDS, and Bloodborne Diseases Department, Pasteur Institute of Iran, Tehran, Iran.

Background: Among the various types of pharmaceuticals, vaccines have a special place. However, in the case of HIV, nearly after 40 years of its discovery, an effective vaccine still is not available. The reason lies in several facts mainly the variability and smartness of HIV as well as the complexity of the interaction between HIV and immune responses. A robust, effective, and longterm immunity is undoubtedly what a successful preventive vaccine should induce in order to prevent the infection of HIV. Failure of human trials to this end has led to the idea of developing therapeutic vaccines with the purpose of curing already infected patients by boosting their immune responses against the virus. Nevertheless, the exceptional ability of the virus to escape the immune system based on the genetically diverse envelope and variable protein products have made it difficult to achieve an efficient therapeutic vaccine.

Objective: We aimed at studying and comparing different approaches to HIV therapeutic vaccines.

Methods: In this review, we summarized the human trials undergoing on HIV therapeutic vaccination which are registered in the U.S. clinical trial database (clinicaltrials.gov). These attempts are divided into different tables, according to the type of formulation and application in order to classify and compare their results.

Result/conclusion: Among several methods applied in studied clinical trials which are mainly divided into DNA, Protein, Peptide, Viral vectors, and Dendritic cell-based vaccines, protein vaccine strategy is based on Tat protein-induced anti-Tat Abs in 79% HIV patients. However, the studies need to be continued to achieve a durable efficient immune response against HIV-1.
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http://dx.doi.org/10.2174/1570162X17666190618160608DOI Listing
February 2020

Naturally occurring NS5A and NS5B resistant associated substitutions in HCV and HCV/HIV patients in iranian population.

Clin Res Hepatol Gastroenterol 2019 10 10;43(5):594-602. Epub 2019 May 10.

Clinical Research Dept, Pasteur Institute of Iran, Tehran, Iran. Electronic address:

Background: The introduction of direct acting antivirals (DAAs) for hepatitis C virus (HCV) treatment promises shorter treatment duration, higher cure rates and fewer side effects. Naturally, occurring Resistance Associated Substitutions (RASs) are major challenge to the success of the HCV antiviral therapy.

Aim: To determine the naturally occurring NS5A and NS5B RASs in Iranian HCV and HCV/human immunodeficiency virus (HIV) patients.

Methods: A total of 209 DAA-naïve chronic HCV patients including 104 HCV mono-infected and 105 HCV/HIV co-infected cases were enrolled. Amplification and Sanger population sequencing of NS5A and NS5B regions of HCV genome were carried out. The amino acid sequence diversity of the NS5A and NS5B regions were analyzed using geno2pheno HCV.

Results: NS5A RASs were detected in 25.5% of HCV and 16.9% of HCV/HIV subjects. In HCV cases, clinically relevant RASs were L28M followed by M28Vand Q30H and Y93H/N. In HCV/HIV subjects, clinically relevant RASs were Y93H/N followed by L28M and P58T and M28V/T and Q30R. NS5B RASs were observed in 11.8% of HCV and 5.9% of HCV/HIV subjects. Clinically relevant substitutions were included V321A/I, C316Y, S282R and L159F. The major S282T mutation was not observed.

Conclusion: The emergence of RASs is a growing issue in the setting of current treatment with DAAs. Although currently, screening of RASs is recommended before specific DAA regimens, it should be consider in patients with therapeutic failure and in the cases of retreatment.
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http://dx.doi.org/10.1016/j.clinre.2019.01.011DOI Listing
October 2019

Significance of serum antibodies against HPV E7, Hsp27, Hsp20 and Hp91 in Iranian HPV-exposed women.

BMC Infect Dis 2019 Feb 12;19(1):142. Epub 2019 Feb 12.

Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran.

Background: Among different types of human papillomavirus (HPV), types 16 and 18 were known to be high-risk agents causing mainly cervical cancer. Up to now, the potential of HPV E7 protein has been proved as a diagnostic marker of cervical cancer. Moreover, the levels of anti-heat shock protein (Hsp) and anti-high mobility group box-1 (HMGB1) antibodies in cancer patients have been useful in tumor diagnosis. The goal of the present study was to determine the efficiency of the potential serologic markers including HPV E7, Hsp20, Hsp27 proteins and Hp91 peptide in Iranian HPV-exposed women, for the first time.

Methods: At first, the recombinant HPV E7, Hsp20 and Hsp27 proteins were expressed in E. coli system, and purified by affinity chromatography under native conditions. Then, antibody responses were detected against the recombinant proteins as well as Hp91 peptide as potential markers in 49 Iranian women who were seropositive for HPV-16 and 18 L1 capsids (i.e., HPV-exposed women) and 49 controls using indirect ELISA.

Results: Our data indicated that the seroreactivities of women exposed to HPV16, HPV18 and both of them against the recombinant E7, Hsp20, Hsp27 proteins and Hp91 peptide were significantly higher than those in control group (p < 0.05 for HPV16 or HPV18; p < 0.01 for both of them versus all markers). HPV-exposed women with high antibody responses to HPV-16 and 18 L1 capsids as a commercial biomarker had significant seroreactivity to HPV-16 and 18 E7 and Hsp27 (p < 0.05). The recombinant E7 and Hsp27 proteins showed higher efficiency than Hsp20 and Hp91 for detection of individuals exposed to HPV infections (p < 0.05).

Conclusion: Generally, the levels of serum E7 and Hsp27 were increased in HPV-16 and 18 L1- seropositive women suggesting their potential value as a diagnostic marker for HPV infections.
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http://dx.doi.org/10.1186/s12879-019-3780-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6373072PMC
February 2019

In Silico Design and Immunologic Evaluation of HIV-1 p24-Nef Fusion Protein to Approach a Therapeutic Vaccine Candidate.

Curr HIV Res 2018 ;16(5):322-337

Hepatitis, AIDS and Bloodborne Diseases Department, Pasteur Institute of Iran, Tehran, Iran.

Background: Acquired immune deficiency syndrome (HIV/AIDS) has been a major global health concern for over 38 years. No safe and effective preventive or therapeutic vaccine has been developed although many products have been investigated. Computational methods have facilitated vaccine developments in recent decades. Among HIV-1 proteins, p24 and Nef are two suitable targets to provoke the cellular immune response. However, the fusion form of these two proteins has not been analyzed in silico yet.

Objective: This study aimed at the evaluation of possible fusion forms of p24 and Nef in order to achieve a potential therapeutic subunit vaccine against HIV-1.

Method: In this study, various computational approaches have been applied to predict the most effective fusion form of p24-Nef including CTL (Cytotoxic T lymphocytes) response, immunogenicity, conservation and population coverage. Moreover, binding to MHC (Major histocompatibility complex) molecules was assessed in both human and BALB/c.

Results: After analyzing six possible fusion protein forms using AAY linker, we came up with the most practical form of p24 from 80 to 231 and Nef from 120 to 150 regions (according to their reference sequence of HXB2 strain) using an AAY linker, based on their peptides affinity to MHC molecules which are located in a conserved region among different virus clades. The selected fusion protein contains seventeen MHC I antigenic epitopes, among them KRWIILGLN, YKRWIILGL, DIAGTTSTL and FPDWQNYTP are fully conserved between the virus clades. Furthermore, analyzed class I CTL epitopes showed greater affinity binding to HLA-B 57*01, HLA-B*51:01 and HLA-B 27*02 molecules. The population coverage with the rate of >70% coverage in the Persian population supports this truncated form as an appropriate candidate against HIV-I virus.

Conclusion: The predicted fusion protein, p24-AAY-Nef in a truncated form with a high rate of T cell epitopes and high conservancy rate among different clades, provides a helpful model for developing a therapeutic vaccine candidate against HIV-1.
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http://dx.doi.org/10.2174/1570162X17666190102151717DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6446525PMC
July 2019

Novel nano-sized chitosan amphotericin B formulation with considerable improvement against Leishmania major.

Nanomedicine (Lond) 2018 12 22;13(24):3129-3147. Epub 2018 Nov 22.

Department of Clinical Research, Pasteur Institute of Iran, Tehran, Iran.

Aim: Improvement in the treatment of Leishmania major's pathological effects through increasing the dose of amphotericin B loaded into nanochitosan.

Materials & Methods: The phase separation method was used for nanochitosan synthesis and amphotericin loading. Also a novel solvent was designed and the nanodrug efficacy was evaluated in vitro and in vivo (pathology) environments.

Results: The drug loading efficiency of 90%, along with slow drug-release with cellular uptake of 98.6% was achieved. The novel solvent was composed of 10% acetic acid, and it was succeeded to dissolve AK10 mg/kg. Also, AK10 mg/kg had no side effects in in vitro and in vivo environments. In addition, the complete wound healing and parasite inhibition were achieved by using AK10 mg/kg in terms of improvement the treatment indicators.

Conclusion: Increasing the therapeutic dose of AK to 10 mg/kg caused the successful treatment of L. major's pathological effects in in vitro and in vivo environments.
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http://dx.doi.org/10.2217/nnm-2018-0063DOI Listing
December 2018

Novel Nanosized Chitosan-Betulinic Acid Against Resistant Leishmania Major and First Clinical Observation of such parasite in Kidney.

Sci Rep 2018 08 6;8(1):11759. Epub 2018 Aug 6.

Department of Clinical Research, Pasteur Institute of Iran, Tehran, Iran.

Regarding the antiparasitic effects of Betulinic acid (B) against Leishmaniasis, it was loaded into nanochitosan (K) for the first time in order to improve its therapeutic effects and decrease its side effects for the treatment of Leishmania major-infected Balb/c mice. Improvement the therapeutic efficacy of Bas an anti-leishmania agent through increasing the effective dose was achieved by using a novel solvent and phase separation method for K synthesis. The synthesized K with the size of 102 nm and Betulinic acid-nanochitosan (BK) with the size of 124 nm and drug loading efficiency of 93%, cellular uptake of 97.5% with the slow drug release pattern was prepared. To increase the therapeutic dose, a modified 10% acetic acid solvent was used. The in vitro and in vivo results showed that the nanodrug of BK was non toxic by 100% and BK20 mg/kg could completely performed the wound healing and inhibit the parasite in a large extent (P ˂ 0.001) compared to other groups. Therefore, BK could be considered as an alternative regimen for treatment of L. major.
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http://dx.doi.org/10.1038/s41598-018-30103-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6078985PMC
August 2018

Reduction toxicity of Amphotericin B through loading into a novel nanoformulation of anionic linear globular dendrimer for improve treatment of leishmania major.

J Mater Sci Mater Med 2018 Jul 28;29(8):125. Epub 2018 Jul 28.

Department of Clinical Research, Pasteur Institute of Iran, Tehran, Iran.

Amphotericin B (A) as an antileishmanial drug has limited clinical application owing to severe side-effects and low-water solubility. This is the first study reported using Anionic Linear Globular Dendrimer (ALGD) as A carrier for the increase of A solubility rate, decrease its toxicity, and improve its therapeutic effects. ALGD was synthesized and A was loaded into nanoparticles for the first time with the drug-loading efficiency of 82%. Drug loading was confirmed using characterization methods. The drug solubility rate was increased by 478-folds. The results of the study showed that the A toxicity was significantly decreased by 95% in vitro and in vivo environments, which was confirmed by pathology findings and enzymatic evaluation. Furthermore, the nanodrug caused that mortality rate was reached to zero. Moreover, the nanodrug was as potent as the free drug and glucantime (GUL) in reducing the parasite burden and parasite number. These findings indicated the potency of ALGD to decrease the drug side-effects, increase the drug solubility rate, and improve the drug efficacy. Moreover, the nanoformulation was a non-toxic and cost-effective formulation. The conformity between in vitro and in vivo results suggested that the A-loaded ALGD could be considered as a promising candidate in reducing the side-effects of A in leishmaniasis treatment.
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http://dx.doi.org/10.1007/s10856-018-6122-9DOI Listing
July 2018

Seroprevalence, molecular epidemiology and quantitation of parvovirus B19 DNA levels in Iranian blood donors.

J Med Virol 2018 08 1;90(8):1318-1322. Epub 2018 May 1.

Department of Clinical Research, Pasteur Institute of Iran, Tehran, Iran.

Human parvovirus B19 (B19) infection is common among blood donors, and healthy blood donors can transmit virus via transfusion. Due to resistance of B19 to viral inactivation methods, there is a potential concern regarding transfusion safety in blood products. We aimed to determine the seroprevalence, molecular epidemiology, and quantitation of B19 DNA levels in blood donors in Tehran, Iran. A total of 500 blood donors from Blood Transfusion Research Center were studied. ELISA was used for detection of B19 IgG and IgM and nested PCR was carried out for detection of B19 DNA. PCR products were subjected to direct sequencing. B19 viral load was determined by real time PCR. B19 IgG, IgM, and DNA were detected in 27.6, 2.6, and 1.2% of donors respectively. Ten samples (2%) were positive for both antibodies while in four cases (0.8%), B19 IgG and DNA detected simultaneously. One case had B19 IgM, IgG, and viremia concurrently. The titers of B19 DNA in four of six donors were more than 10  IU/mL (high level viremia) and all four cases had IgG simultaneously. All B19 isolates categorized in genotype 1A. Our findings indicated that prevalence of B19 DNA in Iranian blood donors was comparable with previous studies throughout the world. High level B19 viremia found in 0.8% of our donors and all viremic donors revealed neutralizing B19 antibody. Therefore implementation of a B19 screening test for each volunteer blood donor does not appear to be necessary but B19 testing for plasma-derived products seems important in Iranian donors.
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http://dx.doi.org/10.1002/jmv.25195DOI Listing
August 2018

Intra-familial Transmission of Hepatitis B Virus Infection in Arak, Central Iran.

Iran J Pathol 2016 ;11(4):328-333

Clinical Research Dept., Pasteur Institute of Iran, Tehran, Iran.

Background: The household transmission of hepatitis B virus (HBV) is a major health problem. High incidence of HBV infection is observed within the household contacts of HBV carriers. We aimed to evaluate serological markers of hepatitis B infection among family members of HBV carriers in Arak, central Iran.

Methods: Data were collected from the 100 chronic HBV carriers (subjects with positive HBsAg for at least 6 months period) as index cases and 700 members of their family. Then, we checked serologic markers of hepatitis B [hepatitis B surface antigen (HBsAg), hepatitis B core antibody (anti-HBc) and hepatitis B surface antibody (anti- HBs)] using the ELISA test.

Results: The prevalence rate of HBsAg, anti-HBs and anti-HBc among household members was 23.3%, 20.4% and 23% respectively. Isolated anti-HBc (positive anti-HBc with negative HBsAg and anti-HBs) found in 0.4% of family members. Mothers and children with 47.6% and 17.2% had the highest and lowest rates of HBV infection, respectively (=0.00). There was a significant difference between mothers and spouses of index case (47.6% and 29.8%) regarding HBsAg positivity (=0.03).

Conclusion: The low rate of HBV infection reported in children reveal the effective prevention of HBV transmission with the universal vaccination programs and also importance of pregnant women screening for HBV serological markers.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5563930PMC
January 2016

No Role of Herpes Simplex Virus Type 2 (HSV-2) Infection on HIV Progression in Naïve HIV Patients

Iran Biomed J 2018 03 9;22(2):123-8. Epub 2017 Jul 9.

Department of Clinical Research, Pasteur Institute of Iran, Tehran, Iran.

Background: Herpes simplex virus type 2 (HSV-2) is a common infection in human immunodeficiency virus (HIV) patients and may accelerate HIV progression by rising HIV viral load and decreasing CD4 count. However, the available data regarding the influence of HSV-2 seropositivity on HIV progression in HIV individuals are inconclusive. Therefore, we aimed to determine HSV-2 seroprevalence in naïve HIV patients and normal controls and also investigate the relation of HIV viral load and CD4 count with HSV-2 seropositivity. Subsequently, we investigated the association of HSV-2 serostatus with changing in CD4 count and HIV viral load in our subjects, after one year follow-up.

Methods: In this study, 116 naïve HIV patients and 85 healthy controls from Tehran, Iran were enrolled. HSV-2 IgG antibody was detected by ELISA. CD4 count was determined by flowcytometry, and serum HIV RNA copy numbers were determined using real-time PCR.

Results: The prevalence of HSV-2 IgG was 18.1% in naïve HIV patients and 0% in the control group (P=0.000). HSV-2 seroconversion was observed in 2.43% of HIV patients after one year. There was no significant difference regarding HSV-2 serostatus with CD4 count and HIV RNA viral load in our study cohort at baseline and after one year.

Conclusion: Our results revealed that the prevalence and incidence of HSV-2 infection are low in our HIV cases, and it is negligible in control group. However, it seems that HIV/HSV2 co-infection has no role on HIV infection acceleration.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5786658PMC
http://dx.doi.org/10.22034/ibj.22.2.123DOI Listing
March 2018

Prevalence and antimicrobial susceptibility of Salmonella and Shigella spp. among children with gastroenteritis in an Iranian referral hospital.

Microb Pathog 2017 Aug 16;109:45-48. Epub 2017 May 16.

Pediatric Infectious Disease Research Center, Tehran University of Medical Science, Tehran, Iran; Department of Infectious Diseases, Pediatrics Center of Excellence, Children's Medical Center, Tehran University of Medical Sciences, Tehran, Iran. Electronic address:

Background: Gastroenteritis is one of the leading cause of illnesses through the world, especially in developing countries.Salmonella and Shigella infections are considered as the main public health problems in children. The aim of this study was to detect the prevalence and antimicrobial susceptibility of Salmonella and Shigella spp. among children with gastroenteritis in an Iranian referral hospital.

Methods: During April 2013 to April 2014, all medical records of children with gastroenteritis admitted to a pediatric medical center were evaluated. Positive stool cultures of children were evaluated and frequency of Salmonella and Shigella spp. and their antimicrobial susceptibility were detected.

Results: In this study, 676 patients with the mean age of 24.94 months were enrolled. Eighty-eight (42%) Salmonella spp., 85 (40%) Shigella spp., 33 (16%) E. coli and 5(2%) candida albicans were isolated from 211 positive stool cultures. Among 85 Shigella spp. isolates, S. sonnei, S. flexneri and other Shigella spp. were isolated from 39 (46%) isolates, 36(42%) and 10(12%), respectively. Among 88 isolated Salmonella spp., 36 (41%) isolates were Salmonella Serogroup D, 26 (30%) were Salmonella Serogroup B, 20 (23%) isolates were Salmonella Serogroup C and 6 (7%) were other Salmonella spp. isolates. Thirty-eight percent of Salmonella serogroup B were resistant to nalidixic acid, while higher frequency of nalidixic acid resistant was found in Salmonella serogroup C and Salmonella serogroup D. The higher frequency of ampicillin resistant was found in Shigella spp. than Salmonella spp. High frequency of cefotaxime resistant was seen in S. sonei and S. flexneri (77% and 56%, respectively), whereas more than 90% of Salmonella serogroup B, C and D were susceptible to this antibiotic.

Conclusion: In conclusion, Shigella and Salmonella serogroups can be considered as important etiological agents of acute diarrhea in children. Since the prevalence of antibiotic resistance is increasing in recent years in Iran, further studies on the prevalence, antimicrobial susceptibility pattern and mechanisms of antibiotic resistance in these species is highly recommended.
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http://dx.doi.org/10.1016/j.micpath.2017.05.023DOI Listing
August 2017

Distribution of hepatitis C virus genotypes in Arak city, central province of Iran.

Iran J Microbiol 2016 Oct;8(5):321-325

Department of Clinical Research, Pasteur Institute of Iran, Tehran, Iran.

Background And Objectives: Hepatitis C virus (HCV) infection is a worldwide concern and it is the major cause of liver disease. Several genotypes of the HCV have been reported from different regions of the world. The determination of the HCV genotypes is important for the prediction of response to antiviral treatment and clinical outcomes. So, HCV genotyping in each region is of great importance. This investigation was performed to determine the distribution of HCV genotypes in Arak city, Central province of Iran.

Patients & Methods: In this cross sectional study, 174 cases with chronic HCV infection from Arak city were enrolled. HCV infection was confirmed by positive results in HCV antibody (anti-HCV) and HCV-RNA tests. HCV genotypes were determined using a PCR based genotyping kit.

Results: A total of 174 HCV infected patients with mean age of 37.5±10.24 years were enrolled. 97.7% of cases were male and 2.3% were female. The main route of HCV transmission was injection drug use (IDU) which was observed in 59.8% of cases. Genotyping results demonstrated that subtype 3a (52.9%) was the most prevalent HCV type in Arak, followed by subtype 1a (22.9%) and subtype 1ab (17.8%).

Conclusion: This study showed that HCV subtype 3a was the most prevalent HCV type, followed by subtype 1a and subtype 1ab in Arak, central province of Iran. Investigation of HCV genotypes in different parts of the country is needed to facilitate treatment options and preventive strategies.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5277601PMC
October 2016

Frequency and Genotype of Human Parvovirus B19 among Iranian Hemodialysis and Peritoneal Dialysis Patients.

Intervirology 2016 1;59(3):179-185. Epub 2017 Feb 1.

Infectious Diseases Research Center, Kashan University of Medical Sciences, Kashan, Iran.

Objectives: The aim of this study was to evaluate the frequency and genotype of human parvovirus B19 and its relation with anemia among Iranian patients under dialysis.

Methods: Fifty hemodialysis (HD) and 33 peritoneal dialysis (PD) patients were enrolled. B19 IgG and IgM antibodies were assessed by ELISA, and the presence of B19 DNA was evaluated by nested PCR. PCR products were sequenced directly and phylogenetic analysis was performed.

Results: In the HD group, the prevalence of B19 antibodies was 54% for IgG and 4% for IgM. B19 DNA was detected in 10% of the cases, and 10% showed B19 IgG and viremia simultaneously. In the PD group, the prevalence of B19 IgG and IgM was 57.6 and 0% respectively, whereas B19 DNA was found in 12.1% of the group. A total of 9.1% showed B19 IgG and viremia concurrently. There was no significant difference regarding anemia and B19 infection in either group. All B19 isolates were clustered in genotype 1A.

Conclusion: Our findings indicate that B19 infection plays no role in leading chronic anemia in dialysis patients. However, persistent B19 viremia and the circulation of the same strains in dialysis patients may indicate a potential risk for the contamination of dialysis equipment and nosocomial spread of B19 infection within dialysis units.
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http://dx.doi.org/10.1159/000455124DOI Listing
March 2017

Interlukine-17 and TGF-β levels in patients with acute brucellosisbefore and after treatment.

Turk J Med Sci 2016 Nov 17;46(5):1348-1352. Epub 2016 Nov 17.

Department of Clinical Research, Pasteur Institute of Iran, Tehran, Iran.

Background/aim: T-helper cell type 1 (Th1)/Th2 cytokine balance is involved in the resistance or susceptibility to Brucella infection. The analysis of cytokine levels is valuable to determine the role of the immune system in Brucella prognosis. The aim of this study was to investigate the levels of serum interleukin-17 (IL-17) and transforming growth factor beta (TGF-β) and their alterations with treatment in patients with acute brucellosis.

Materials And Methods: TGF-β was tested in 33 acute brucellosis patients and 19 controls and IL-17 was analyzed in 40 patients and 12 controls. Cytokine levels were tested in controls and patients before and after treatment by ELISA.

Results: TGF-β levels were significantly lower in brucellosis cases compared to controls. At the end of the treatment, the serum levels of this cytokine had increased, but there was no significant difference between this cytokine level before and after treatment. The IL-17 level was significantly higher in the brucellosis group compared to controls and its value decreased in patients at the end of treatment without any significant difference.

Conclusion: This study indicated that TGF-β was lower and IL-17 was higher in brucellosis cases and, after treatment, the serum level of TGF-β increased and that of IL-17 decreased in these patients.
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http://dx.doi.org/10.3906/sag-1506-59DOI Listing
November 2016

Gender and age-specific seroprevalence of human papillomavirus 16 and 18 in general population in Tehran, Iran.

Med Microbiol Immunol 2017 Apr 17;206(2):105-110. Epub 2016 Nov 17.

Clinical Research Department, Pasteur Institute of Iran, No 69, Pasteur Ave., Tehran, 13164, Iran.

The assessment of the gender and age-specific seroprevalence of human papillomavirus (HPV) is essential for planning of HPV vaccine implementation into the preventive programs. In this study, we aimed to determine the age-specific seroprevalence of HPV-16 and 18 in both males and females in Tehran, Iran. Three hundred and seventy-eight women (10-35 years) and 162 men (10-25 years) from Tehran, Iran, were enrolled. Anti-HPV IgG antibodies against HPV-16 and HPV-18 were detected by ELISA using papillomavirus type 16 and 18 L1-capsids as antigen. HPV-16 antibody was detected in 15.6 and 13.6% of women and men, respectively. Antibody against HPV-18 was found positive in 12.7 and 8% of women and men, respectively. The highest seroprevalence of HPV-16 and 18 were seen in women aged 26-30 years (22.2 and 19.4%, respectively), and the lowest HPV-16 and 18 seropositivity rates were seen in males and females aged 10-15 years (9.3 and 1.9%, respectively). In our cohort of study, in males, both anti-HPV-16 and 18 increased after age 15 years, peaking in men aged 21-25 years. In women, both HPV-16 and 18 seropositivity increased after 15 years, declined at 21-25 years, peaked in women aged 26-30 years and again decreased after 30 years. Our data showed increasing exposure rate to high-risk HPV vaccine types in our studied population over 15 years of age. In order to prevent the HPV-related cancers, implementation of HPV vaccine into the national immunization program in Iran and vaccination of females and males less than 15 years of age are suggested.
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http://dx.doi.org/10.1007/s00430-016-0487-5DOI Listing
April 2017

BK Viremia among Iranian Renal Transplant Candidates.

Iran J Pathol 2016 ;11(3):210-215

Clinical Research Dept., Pasteur Institute of Iran, Tehran, Iran.

Background: Primary infection with BK virus (BKV) is occurred during childhood and usually asymptomatic, but after initial infection, BKV may persist lifelong in the kidney and genitourinary tract. Reactivation may occur in individuals with compromised immunity such as renal transplant recipients. Due to the role of BKV in BK virus-associated nephropathy (BKVAN) and potentially renal allograft rejection, the detection of BKV in renal transplant candidates is very important. The aim of this study was to evaluate the frequency of BK viremia in end stage renal disease cases who were candidates for renal transplantation.

Methods: In this cross-sectional study, 50 cases with end stage renal disease who were candidates for renal transplantation were recruited from the main dialysis unit in Tehran, Iran. Presence of BK viremia was determined in plasma samples of cases using real time PCR.

Results: A total of 50 renal transplant candidates with mean age 37.8±13 yr were enrolled in the study. Fifty two percent of subjects were male. Forty six (92%) of them were under HD and 4 (8%) were on PD. BK virus was not detected in any plasma samples of renal transplant candidates.

Conclusion: This study showed absence of BK viremia in our renal transplant candidates. However, due to the important role of BKV in BKVAN and renal graft failure and rejection, further studies involving larger number of cases are required to elucidate the rate of the BKV in renal transplant candidates.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5079453PMC
January 2016