Publications by authors named "Seyed Alireza Nadji"

51 Publications

Clinical Manifestations of Patients with Coronavirus Disease 2019 (COVID-19) in a Referral Center in Iran.

Tanaffos 2020 Nov;19(2):122-128

Clinical Tuberculosis and Epidemiology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Background: Following the recent epidemic of coronavirus disease 2019 (COVID-19) in Wuhan, China, a novel betacoronavirus was isolated from two patients in Iran on February 19, 2020. In this study, we aimed to determine the clinical manifestations and outcomes of the first confirmed cases of COVID-19 infection (n=127).

Materials And Methods: This prospective study was conducted on all COVID-19-suspected cases, admitted to Masih Daneshvari Hospital (a designated hospital for COVID-19), Tehran, Iran, since February 19, 2020. All patients were tested for COVID-19, using reverse transcription-polymerase chain reaction (RT-PCR) assay. Data of confirmed cases, including demographic characteristics, clinical features, and outcomes, were collected and compared between three groups of patients, requiring different types of admission (requiring ICU admission, admission to the general ward, and transfer to ICU).

Results: Of 412 suspected cases, with the mean age of 54.1 years (SD=13.4), 127 (31%) were positive for COVID-19. Following the patients' first visit to the clinic, 115 cases were admitted to the general ward, while ten patients required ICU admission. Due to clinical deterioration in the condition of 25 patients (out of 115 patients), ICU admission was essential. Based on the results, the baseline characteristics of the groups were similar. Patients requiring ICU admission were more likely to have multiorgan involvement (liver involvement, P<0.001; renal involvement, P<0.001; and cardiac involvement, P=0.02), low O saturation (P<0.001), and lymphopenia (P=0.05). During hospital admission, 21 (16.5%) patients died, while the rest (83.5%) were discharged and followed-up until March 26, 2020. Also, the survival rate of patients, who received immunoglobulin, was higher than other patients (60.87% vs. 39.13%).

Conclusion: The mortality rate of COVID-19 patients was considerable in our study. Based on the present results, this infection can cause multiorgan damage. Therefore, intensive monitoring of these patients needs to be considered.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7680520PMC
November 2020

Effects of Exogenous Melatonin on MAM Induced Lung Injury and Lung Development in Mice Offspring.

Tanaffos 2020 Jan;19(1):66-73

Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

Background: Melatonin as an antioxidant agent can have an effective role in lung development. In this study, the effect of melatonin administration on lung injury in the neonate mice was assessed.

Materials And Methods: Lung injury was induced by two injections of 15 mg/kg methylazoxymethanol (MAM) on gestational day 15 (E15). Pregnant BALB/c mice were randomly divided into five groups: Control (CO), Melatonin (MEL), Luzindole (Luz), MAM, and MAM+MEL. Melatonin and luzindole were intra-peritoneally injected at a dose of 10 mg/kg (from E15 until delivery). Histopathological changes including: hemorrhage, neutrophils infiltration and fibrosis in the neonate lung were studied by hematoxylin and eosin (H&E) and Masson's Trichrome staining. Alveolarization and alveolar wall thickness were measured.

Results: In histological examination, hemorrhage, neutrophils infiltration and fibrosis were seen in the MAM and Luz groups; however, these injuries were attenuated in the MAM plus melatonin group. Significant reduction of alveolarization was recorded in the MAM and Luz groups compared to the control group, while the alveolar wall thickness was significantly increased in these groups compared to control group.

Conclusion: Administration of exogenous melatonin in pregnant mice could have a protective effect on the pulmonary development of neonates and could decrease lung injury in neonate mice.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7569496PMC
January 2020

Frequent detection of enterovirus D68 and rhinovirus type C in children with acute respiratory infections.

Eur J Clin Microbiol Infect Dis 2021 Mar 3;40(3):637-642. Epub 2020 Oct 3.

Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, 1471613151, Iran.

This study aimed to evaluate the prevalence of human rhinoviruses (HRVs) and the emergence of enterovirus D68 (EV-D68) in children. A total of 322 nasopharyngeal swab samples were provided from children with an initial diagnosis of upper and lower respiratory tract infections. A total of 34 and 70 cases were positive for EV-D68 and HRV, respectively. The phylogenetic analysis revealed that the clades A and B are the prevalent genotypes for EV-D68 and the HRV-positive samples belong to three types including HRV-A, HRV-B, and HRV-C. The results showed that EV-D68 and HRV-C are circulating in Iran especially in the winter.
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http://dx.doi.org/10.1007/s10096-020-04051-yDOI Listing
March 2021

Diffuse alveolar damage and thrombotic microangiopathy are the main histopathological findings in lung tissue biopsy samples of COVID-19 patients.

Pathol Res Pract 2020 Oct 19;216(10):153228. Epub 2020 Sep 19.

Chronic Respiratory Diseases Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Background: Since the outbreak of the novel coronavirus disease-2019 (COVID-19) in December 2019, limited studies have investigated the histopathologic findings of patients infected with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2).

Material And Methods: This study was conducted on 31 deceased patients who were hospitalized for COVID-19 in a tertiary hospital in Tehran, Iran. A total of 52 postmortem tissue biopsy samples were obtained from the lungs and liver of decedents. Clinical characteristics, laboratory data, and microscopic features were evaluated. Reverse transcription polymerase chain reaction (RT-PCR) assay for SARS-CoV-2 was performed on specimens obtained from nasopharyngeal swabs and tissue biopsies.

Results: The median age of deceased patients was 66 years (range, 30-87 years) and 25 decedents (81 %) were male. The average interval from symptom onset to death was 13 days (range, 6-34 days). On histopathologic examination of the lung specimens, diffuse alveolar damage and thrombotic microangiopathy were the most common findings (80 % and 60 %, respectively). Liver specimens mainly showed macrovesicular steatosis, portal lymphoplasmacytic inflammation and passive congestion. No definitive viral inclusions were observed in any of the specimens. In addition, 92 % of lung tissue samples tested positive for SARS-CoV-2 by RT-PCR.

Conclusions: Further studies are needed to investigate whether SARS-CoV-2 causes direct cytopathic changes in various organs of the human body.
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http://dx.doi.org/10.1016/j.prp.2020.153228DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7837112PMC
October 2020

Promising effects of tocilizumab in COVID-19: A non-controlled, prospective clinical trial.

Int Immunopharmacol 2020 Nov 4;88:106869. Epub 2020 Aug 4.

Clinical Tuberculosis and Epidemiology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Masih Daneshvari Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Electronic address:

Background: The clinical presentation of SARS-CoV-2 infection ranges from mild symptoms to severe complications, including acute respiratory distress syndrome. In this syndrome, inflammatory cytokines are released after activation of the inflammatory cascade, with the predominant role of interleukin (IL)-6. The aim of this study was to evaluate the effects of tocilizumab, as an IL-6 antagonist, in patients with severe or critical SARS-CoV-2 infection.

Methods: In this prospective clinical trial, 76 patients with severe or critical SARS-CoV-2 infection were evaluated for eligibility, and ultimately, 42 patients were included. Tocilizumab was administered at a dose of 400 mg as a single dose via intravenous infusion. Primary outcomes included changes in oxygenation support, need for invasive mechanical ventilation, and death. Secondary outcomes included radiological changes in the lungs, IL-6 plasma levels, C-reactive protein levels, and adverse drug reactions. The data were analyzed using SPSS software.

Results: Of the 42 included patients, 20 (48%) patients presented the severe infection stage and 22 (52%) were in the critical stage. The median age of patients was 56 years, and the median IL-6 level was 28.55 pg/mL. After tocilizumab administration, only 6 patients (14%) required invasive ventilation. Additionally, 35 patients (83.33%) showed clinical improvement. By day 28, a total of 7 patients died (6 patients in the critical stage and 1 patient in the severe stage). Neurological adverse effects were observed in 3 patients.

Conclusions: Based on the current results, tocilizumab may be a promising agent for patients with severe or critical SARS-CoV-2 infection, if promptly initiated during the severe stage.
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http://dx.doi.org/10.1016/j.intimp.2020.106869DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7402206PMC
November 2020

Subcutaneous administration of interferon beta-1a for COVID-19: A non-controlled prospective trial.

Int Immunopharmacol 2020 Aug 7;85:106688. Epub 2020 Jun 7.

Clinical Tuberculosis and Epidemiology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran. Electronic address:

Background: Recently, a new coronavirus spreads rapidly throughout the countries and resulted in a worldwide epidemic. Interferons have direct antiviral and immunomodulatory effects. Antiviral effects may include inhibition of viral replication, protein synthesis, virus maturation, or virus release from infected cells. Previous studies have shown that some coronaviruses are susceptible to interferons. The aim of this study was to evaluate the therapeutic effects of IFN-β-1a administration in COVID-19.

Methods: In this prospective non-controlled trial, 20 patients included. They received IFN-β-1a at a dose of 44 µg subcutaneously every other day up to 10 days. All patients received conventional therapy including Hydroxychloroquine, and lopinavir/ritonavir. Demographic data, clinical symptoms, virological clearance, and imaging findings recorded during the study.

Results: The mean age of the patients was 58.55 ± 13.43 years. Fever resolved in all patients during first seven days. Although other symptoms decreased gradually. Virological clearance results showed a significant decrease within 10 days. Imaging studies showed significant recovery after 14-day period in all patients. The mean time of hospitalization was 16.8 ± 3.4 days. There were no deaths or significant adverse drug reactions in the 14-day period.

Conclusions: Our findings support the use of IFN-β-1a in combination with hydroxychloroquine and lopinavir/ritonavir in the management of COVID-19.

Clinical Trial Registration Number: IRCT20151227025726N12.
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http://dx.doi.org/10.1016/j.intimp.2020.106688DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7275997PMC
August 2020

Tocilizumab administration in a refractory case of COVID-19.

Int J Antimicrob Agents 2020 Aug 2;56(2):106043. Epub 2020 Jun 2.

Clinical Tuberculosis and Epidemiology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran. Electronic address:

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http://dx.doi.org/10.1016/j.ijantimicag.2020.106043DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7264941PMC
August 2020

Association of specific viral infections with childhood asthma exacerbations.

Interv Med Appl Sci 2019 Mar;11(1):17-20

Mycobacteriology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Masih Daneshvari Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Introduction: Asthma exacerbations may occur due to a variety of triggers including respiratory viruses. The aim of this study was to determine the role of particular viral infections in asthma exacerbations in children.

Materials And Methods: The study was performed at Dr. Daneshvari Hospital Pediatric Emergency Department, Shahid Beheshti University of Medical Sciences, Tehran, Iran between 2014 and 2015. A nasopharyngeal aspirate or swab was obtained from each patient during admission. All samples were maintained at 4 °C until submission to the virology laboratory and were tested for respiratory viruses by nucleic acid testing.

Results: A total of 60 patients with asthma exacerbations were recruited for this study. Of the 60 samples collected from the patients with acute asthma exacerbations, rhinovirus was detected in 12 patients (20%), respiratory syncytial virus in 5 (8%), adenovirus in 5 (8%), and influenza virus in 1 (1.6%). Respiratory pathogens were not detected in 37 (61%) samples. All the samples investigated showed single viral infection.

Conclusions: To conclude, the most common viruses detected were rhinovirus followed by respiratory syncytial virus (RSV) and adenovirus. RSV was more commonly associated with more severe attacks. Both the study design (e.g., time of sampling, age of the patients, etc.) and also the method used for viral detection influence the frequency of detection of the respiratory viruses.
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http://dx.doi.org/10.1556/1646.10.2018.35DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7044566PMC
March 2019

Declining trends in HIV and other sexually transmitted infections among female sex workers in Iran could be attributable to reduced drug injection: a cross-sectional study.

Sex Transm Infect 2020 02 13;96(1):68-75. Epub 2019 Dec 13.

Social Determinants of Health Research Center, Institute for Futures Studies in Health, Kerman University of Medical Sciences, Kerman, Iran.

Objective: The HIV trend among female sex workers (FSWs) is understudied. We assessed the prevalence and trend of HIV and five other STIs among FSWs in Iran.

Methods: We recruited FSWs (1337 in 2015, 1005 in 2010) from 21 sites in 13 cities in two cross-sectional biobehavioural surveys. Eligible FSWs were women aged ≥18 years who reported selling sex to more than one male client in the past 12 months. Consenting FSWs were interviewed using a behavioural questionnaire and tested for HIV and five other STIs. We considered study sites as clusters in the analysis and two-sided Fisher's exact test to compare the HIV prevalence between the two survey rounds.

Results: HIV prevalence was 2.1% in 2015 (vs 4.0% in 2010, p=0.007). Lifetime drug injection was reported by 6.1% of participants in 2015 (vs 14.6% in 2010, p=0.003). In 2015, among FSWs with history of lifetime drug injection, HIV prevalence was 8.6% (vs 9.8% in 2010, p=0.425). The prevalence of other STIs in 2015 was 0.4% (95% CI 0.2 to 1.0) for syphilis, 1.3% (95% CI 0.8 to 2.1) for gonorrhoea, 6.0% (95% CI 4.8 to 7.4) for chlamydia, 11.9% (95% CI 8.5 to 16.5) for trichomoniasis and 41.8% (95% CI 39.2 to 44.5) for human papillomavirus.

Conclusions: HIV prevalence among FSWs in Iran decreased, but remains considerably high. The decrease in HIV prevalence compared with 2010 might be explained by a decrease in drug injection. Other STIs are also high in this population. Harm reduction programmes need to be continued and scaled up among this underserved population in Iran.
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http://dx.doi.org/10.1136/sextrans-2018-053800DOI Listing
February 2020

Absence of in Benign and Malignant Breast Tissue.

Iran J Pathol 2019 22;14(4):279-283. Epub 2019 Sep 22.

Department of Pathology, Pathobiology Laboratory Center, Tehran, Iran.

Background & Objective: Malignant breast tumors, which are one of the most important deadly cancers in women, like many other cancers, are proposed to be related to viruses etiologically. Proper management of breast carcinoma necessitates an identification of the etiological factors. is considered to have an etiological role in breast carcinoma. We carried out this study to find out if -DNA is present in the malignant and benign breast tissue in our patients.

Methods: Seventy-five paraffin-embedded breast cancer tissues and 75 normal breast tissues and benign breast lesions were examined in this study (case-control) to look for -DNA employing Nested Polymerase Chain reaction. The tissues were examined over a period of ten years in the pathology department of the Pathobiology Laboratory Center of Tehran.

Results: No -DNA was found in any of the malignant or control group specimens.

Conclusion: Our results showed no evidence of Human Papillomavirus in cancerous and benign tissues, which is consistent with some other studies in English medical literature. More investigations using more specimens from different parts of the country are required to confirm the presence or absence of any connection between Human Papillomavirus and development of breast carcinoma in Iran.
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http://dx.doi.org/10.30699/ijp.2019.89684.1847DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6824765PMC
September 2019

Enterovirus-Human Rhinovirus as a Leading Cause of ARDS in a Liver Transplant Recipient.

Tanaffos 2019 Feb;18(2):169-172

Clinical Tuberculosis and Epidemiology Research Center, NRITLD, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

A 35- year- old man with a prior history of liver transplantation 18 months ago was admitted to our Intensive Care Unit (ICU) with fever and worsening dyspnea and was diagnosed with severe pneumonia leading to Acute Respiratory Distress Syndrome (ARDS). He had a prolonged hospitalization and was treated with empiric broad spectrum intravenous antibiotics, oseltamivir, trimethoprim/sulfamethoxazole, and subsequently caspofungin and ganciclovir. Blood, nasopharyngeal, as well as Bronchoalveolar Lavage (BAL) culture and Polymerase Chain Reaction (PCR) were negative for all viral, bacterial, and fungal causes of pulmonary infection except Enterovirus-Human Rhinovirus (EV-HRV) that was positive with high titers on BAL and swab specimens. Consequently, the diagnosis of EV-HRV pneumonia complicated by ARDS was established. The patient gradually improved and was discharged from the hospital after 3 weeks. This report highlights EV-HRV as a cause of ARDS in immunocompromised adults.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7230127PMC
February 2019

Prevalence of cytomegalovirus infection in patients with ulcerative colitis: a prospective cross-sectional study in Tehran, Iran.

Iran J Microbiol 2018 Oct;10(5):342-347

Department of Pathology, Milad Hospital, Tehran, Iran.

Background And Objectives: Cytomegalovirus (CMV) infection has been reported in ulcerative colitis (UC), but limited data are available on its prevalence in Iran. The aim of this study was to evaluate the prevalence of CMV infection in patients with UC.

Materials And Methods: A prospective, cross-sectional study was conducted in 86 consecutive patients with UC. Prevalence of CMV infection was determined by rectal biopsies for hematoxylin and eosin staining and PCR. CMV-positive specimens was measured for CMV loads by real-time PCR assay.

Results: In six out of 86 (7%) patients with UC, CMV was diagnosed. These patients had detectable CMV DNA in their biopsies as indicated by PCR. In all CMV-positive patients, viral load was more than 250 copy/mg. Histochemical staining did not show any CMV inclusion bodies. No significant demographic and clinical differences existed between patients with and without a CMV infection.

Conclusion: UC and its treatment may put patients at risk of CMV infection. Real-time PCR test for the detection of CMV in UC patients may enable diagnosis of CMV infection with a high sensitivity and allow effective treatment to be administered in these patients. The impact of antiviral therapy on the clinical outcome of the UC patients with CMV remains to be elucidated.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6339997PMC
October 2018

Comparison of a new in-house HIV-1 TaqMan real-time PCR and three commercial HIV-1 RNA quantitative assays.

Comp Immunol Microbiol Infect Dis 2018 Aug 11;59:1-7. Epub 2018 Sep 11.

Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Department of Biotechnology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Electronic address:

Objectives: The aim of this study was to compare the analytical performance of an In-House HIV-1 viral load determination technique with three commercial kits including COBAS AmpliPrep, RealStar, and RTA HIV-1 Real-Time PCR.

Results: A total of 100 HIV-1 suspicious plasma samples were tested by the In-House TaqMan Real-Time PCR assay along with the above-mentioned kits. Comparative analysis between In-House and reference method (COBAS AmpliPrep/COBAS TaqMan HIV-1 Test version 2.0) showed high concordance with a mean difference of 0.08 log copies/ml. All samples results were within -0.16-0.31 log copies/ml. A suitable correlation was obtained with a coefficient (R) of 0.82 between the In-House assay and RTA Kit, however, two positive samples were not detected. The lowest agreement was detected with RealStar HIV Kit 1.0 (R = 0.49, r = 0.7).

Conclusions: The newly developed method has suitable sensitivity, accuracy, and precision. In addition, it is cost-effective and can be an alternative in all laboratories.
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http://dx.doi.org/10.1016/j.cimid.2018.09.002DOI Listing
August 2018

Small Heat Shock Proteins B1 and B6: Which One is the Most Effective Adjuvant in Therapeutic HPV Vaccine?

IUBMB Life 2018 10 1;70(10):1002-1011. Epub 2018 Sep 1.

Virology Research Center (VRC), National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Therapeutic human papillomaviruse (HPV) vaccines have the potential to inhibit the tumor growth by targeting HPV E6 and E7 oncoproteins. Among different vaccine strategies, DNA and protein-based approaches are the most effective candidates for stimulation of the immune responses against HPV infections. Our study was designed to assess the efficacy of small heat shock proteins B1 (Hsp27) and B6 (Hsp20) as an adjuvant accompanied by HPV16 E7 and hPP10-E7 antigens in tumor mouse model. A major key for successful DNA and protein transfer into cells is the development of delivery systems with high efficiency and low cytotoxicity. Herein, we used hPP10 and MPG cell penetrating peptides (CPPs) for protein and DNA delivery in vivo, respectively. Our data indicated that the combination of Hsp27 with the recombinant hPP10-E7 protein in homologous protein/protein (hPP10-E7 + Hsp27) and heterologous DNA/protein (pcDNA-E7 + MPG/ hPP10-E7 + Hsp27) significantly enhanced the E7-specific T cell responses. Indeed, these regimens induced high levels of IgG2a, IFN-γ and IL-2 directed toward Th1 responses and also Granzyme B secretion as compared to other immunization strategies, and also displayed complete protection more than 60 days after treatment. These data suggest that the use of Hsp27 as an adjuvant and MPG and hPP10 as a gene and protein carrier would represent promising applications for improvement of HPV therapeutic vaccines. © 2018 IUBMB Life, 70(10):1002-1011, 2018.
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http://dx.doi.org/10.1002/iub.1892DOI Listing
October 2018

Symptom-Based Versus Laboratory-Based Diagnosis of Five Sexually Transmitted Infections in Female Sex Workers in Iran.

AIDS Behav 2018 Jul;22(Suppl 1):19-25

HIV/STI Surveillance Research Center, and WHO Collaborating Center for HIV Surveillance, Institute for Futures Studies in Health, Kerman University of Medical Sciences, Kerman, Iran.

Among 1337 Iranian adult female sex workers in 2015, we assessed the diagnostic value of 4 self-reported sexually transmitted infection (STIs) symptoms for detecting laboratory-confirmed gonorrhea, chlamydia, trichomoniasis, human papillomavirus (HPV), and syphilis. While 37.7% reported vaginal discharge (VD), 25.9% reported pain or burning (P/B), 3.0% reported genital ulcers (GU), and 1.4% reported genital warts (GW), the prevalence of laboratory-confirmed syphilis, gonorrhea, chlamydia, trichomoniasis, and HPV was 0.4, 1.3, 6.0, 11.9, and 41.9%, respectively. The sensitivity of VD was 40.3% for detecting tricomoniasis, 37.5% for chlamydia, and 37.5% for gonorrhea. The sensitivity of P/B ranged from 12.5% for gonorrhea to 25.2% for trichomoniasis. The sensitivity of GU and GW was very low for 5 STIs. The sensitivity of all symptoms combined was also lower than 50%. Among asymptomatic participants, 41.2% tested positive for HPV, 11.8% for trichomoniasis, and less than 6.6% for other STIs. Symptom-based case management and surveillance of STIs can lead to misclassification of a large proportion of cases.
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http://dx.doi.org/10.1007/s10461-018-2130-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6226376PMC
July 2018

Delivery of molecular cargoes in normal and cancer cell lines using non-viral delivery systems.

Biotechnol Lett 2018 Jun 9;40(6):923-931. Epub 2018 Apr 9.

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

Objective: In this study, transfection efficiency of human papillomavirus (HPV) E7 DNA and protein constructs into HEK-293T normal cell line, and A549 and TC-1 tumor cell lines was evaluated by four delivery systems including supercharge GFP, hPP10 cell penetrating peptide, TurboFect and Lipofectamine using fluorescence microscopy and flow cytometry.

Results: The results indicated that Lipofectamine 2000 and TurboFect produced more effective transfection for GFP and E7-GFP DNA constructs in HEK-293T cells compared to in A549 and TC-1 cells (p < 0.05). In contrast, the supercharge GFP was efficient for E7 DNA and E7 protein delivery in both normal cell (~ 83.94 and ~ 77.01% for HEK-293T), and cancer cells (~ 71.69 and ~ 67.19% for TC-1, and ~ 73.86 and ~ 67.49% for A549), respectively. Indeed, in these cell lines, transfection efficiency by +36 GFP reached ~ 60-80%. Moreover, the hPP10 produced the best transfection result for E7-GFP protein in HEK-293T cells (~ 63.66%) compared to TurboFect (~ 32.95%); however, the efficiency level of hPP10 was only ~ 17.51 and ~ 16.36% in TC-1 and A549 cells.

Conclusions: Our data suggested that the supercharge GFP is the most suitable transfection vehicle for DNA and protein delivery into TC-1 and A549 tumor cell lines compared to other carriers.
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http://dx.doi.org/10.1007/s10529-018-2551-2DOI Listing
June 2018

Therapeutic effect of perinatal exogenous melatonin on behavioral and histopathological changes and antioxidative enzymes in neonate mouse model of cortical malformation.

Int J Dev Neurosci 2018 08 29;68:1-9. Epub 2018 Mar 29.

Institute of Neuroanatomy, RWTH Aachen University, Aachen, Germany; Giulan Neuroscience Research Center, Department of Anatomical Sciences, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran. Electronic address:

Background: Melatonin, which is an antioxidant and neuroprotective agent, can be an effective treatment for neurological disorders. We assessed the effect of melatonin administration on histological changes, antioxidant enzyme levels, and behavioral changes in a neonate mouse model of cortical malformation.

Materials And Methods: Cortical malformation was induced by two injections of 15 mg/kg methylazoxymethanol (MAM) on gestational day 15 (E15). Pregnant Balb/c mice were randomly divided into the following six groups: Control (CO), Melatonin (MEL), Luzindole (LUZ), MAM, MEL + MAM1 (co-treatment), and MEL + MAM2 (pretreatment). Melatonin was intraperitoneally injected at a dose of 10 mg/kg daily (from E15 until delivery of from E6 for 20 days after delivery). On postnatal day 31, the activity and anxiety of mice were assessed by open field and elevated plus maze tests, respectively. Histopathological changes in the neonate cortex were studied using hematoxylin and eosin staining and neurofilament immunohistochemistry. Enzyme-linked immunosorbent assays were used to measure the activity of nitric oxide (NO), malondialdehyde (MDA), and antioxidant enzymes, including catalase (CAT), super oxide dismutase (SOD), and glutathione peroxidase (GPX).

Results: In the behavioral assessment of neonate mice, a significant increase in the crossing activity and decrease in anxiety were recorded in groups treated with MAM plus melatonin. In histological examination, heterotopic, dysmorphic, and ectopic cells, as well as dyslamination, were seen in the MAM and LUZ groups. However, these defects were attenuated in the MAM plus melatonin groups. Significant reductions were recorded in the SOD and GPX levels in the MAM and LUZ groups compared to the control, while the NO level was increased in these groups. Groups that received MAM plus melatonin showed significant increases in the levels of SOD and GPX and a significant decrease in the level of NO, compared to the MAM group.

Conclusion: Melatonin increased the crossing activity and decreased the anxiety in the treated mice of the neonate mouse model of cortical malformation. Histologically, the administration of exogenous melatonin in pregnant mice and their neonates had a protective effect on the cerebral cortex of neonates. Also, this effect is elicited by decreasing NO and increasing antioxidative enzymes.
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http://dx.doi.org/10.1016/j.ijdevneu.2018.03.008DOI Listing
August 2018

and Langerhans Cell Histiocytosis.

Iran J Pathol 2017 1;12(4):323-328. Epub 2017 Nov 1.

Pediatric Infections Research Center, Research Institute for Children Health, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Background And Objective: Langerhans cell histiocytosis (LCH) is a rare histiocytic proliferative disorder of unknown etiology and mainly affects young children. The histological feature is granuloma-like proliferation of langerhans-type dendritic cells. Although the possible role of viruses such as (, ), (), () types 1 and 2 and (CMV, ) is suggested in the pathogenesis of LCH by some investigators, its exact pathophysiology has not been cleared yet. In this study, we investigated the presence of types 1 and 2 in Iranian children with LCH.

Methods: In this retrospective study, we investigated the prevalence of presence of types 1 and 2 (in 30 patients with LCH), using paraffin-embedded tissue samples and 30 age and tissue-matched controls (operated for reasons other than infectious diseases) from the Department of Pediatric Pathology, Tehran, Iran, by nested Polymerase Chain reaction method. No ethical issues arose in the study, because only the pathology reports were reviewed and patients were anonymous.

Results: We failed to find types 1 and 2 DNA in any of the 30 patients with LCH or the control group.

Conclusion: According to our findings, types 1 and 2 do not appear to have any etiologic role in the pathogenesis of LCH in Iranian children. These results are in accordance with previous investigations with negative findings.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5844676PMC
November 2017

New design, development, and optimization of an in-house quantitative TaqMan Real-time PCR assay for HIV-1 viral load measurement.

HIV Clin Trials 2018 04 23;19(2):61-68. Epub 2018 Feb 23.

e Cellular and Molecular Biology Research Center , Shahid Beheshti University of Medical Sciences , Tehran , Iran.

Background Viral load measurement is commonly applicable to monitor HIV infection in patients to determine the number of HIV-RNA in serum samples of individuals. The aim of the present study was to set up a highly specific, sensitive, and reproducible home-brewed Real-time PCR assay based on TaqMan chemistry to quantify HIV-1 RNA genome. Methods In this study, three sets of primer pairs and a TaqMan probe were designed for HIV subtypes conserved sequences. An internal control was included in this assay to evaluate the presence of inhibition. Standard curve and threshold cycle values were determined using in vitro transcribed RNA from int region of HIV-1. A serial dilution of RNA standards was generated by in vitro transcription, from 10 to 10 copies/ml to find the sensitivity and the limit of detection (LOD) of the assay and to evaluate its performance in a quantitative RT-PCR assay. Results The assay has a low LOD equivalent to 33.13 copies/ml of HIV-1 RNA and a linear range of detection from 10 to 10 copies/ml. The coefficient of variation (CV) for Inter and Intra-assay precision of this in-house HIV Real-time RT-PCR ranged from 0.28 to 2.49% and 0.72 to 4.47%, respectively. The analytical and clinical specificity was 100%. Conclusions The results indicate that the developed method has a suitable specificity and sensitivity and is highly reproducible and cost-benefit. Therefore, it will be useful to monitor HIV infection in plasma samples of individuals.
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http://dx.doi.org/10.1080/15284336.2018.1440991DOI Listing
April 2018

Neuraminidase Gene Variations in Influenza A(H1N1)pdm09 Virus among Patients Admitted to Refferal Pulmonary Hospital, Tehran, Iran in 2009-2013.

Tanaffos 2017 ;16(2):99-106

Virology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Background: Neuraminidase (NA) is one of the surface proteins of influenza A virus, which plays an important role in immunization against influenza infection and is recognized as an important therapeutic target. Genetic and antigenic changes and substitutions can influence the efficacy of vaccine and change viral sensitivity to NA inhibitors (NAIs). In this study, we performed phylogenetic and molecular analyses of NA changes in influenza A(H1N1)pdm09 virus, compared them with the corresponding vaccine strain, and examined drug resistance mutations in isolates from patients.

Materials And Methods: The complete sequence of NA genes from 34 pandemic H1N1 isolates (identified in 2009-2010, 2010-2011, and 2013) was determined and analyzed both genetically and antigenically. The phylogenetic tree was plotted relative to the corresponding vaccine strain, using MEGA6 software package, based on the maximum likelihood method and JTT matrix (bootstrap value of 1000).

Results: The phylogenetic analysis of pandemic isolates showed 31 amino acid substitutions in NA genes, compared to the vaccine strain. Some of these substitutions (N248D, V241I, N369K, N44S, and N200S) were important in terms of phylogenetic relationship, while the rest (D103N, V106I, R130T, N200S, G201E, and G414R) influenced the antigenic indices of B-cell epitopes. The catalytic sites, framework sites, and N-glycosylation remained unchanged in the studied samples. Meanwhile, H275Y substitution, related to oseltamivir resistance, was detected in 3 isolates. The average nucleotide identity of NAs with the corresponding vaccine strain was 99.415%, 98.607%, and 98.075% in 2009-2010, 2010-2011, and 2012-2013, respectively.

Conclusion: In this study, we provided basic information on the genetic and antigenic changes of NA genes in influenza A(H1N1)pdm09 virus from patients in 3 different seasons in Tehran, Iran. Considering the viral NAI resistance and changes in NA gene sequences of the isolates in comparison with the vaccine strain, further studies should be performed to monitor genetic changes in Iran. Moreover, the efficacy of vaccines should be examined.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5749334PMC
January 2017

Lack of HPV in Benign and Malignant Epithelial Ovarian Tumors in Iran

Asian Pac J Cancer Prev 2017 05 1;18(5):1233-1236. Epub 2017 May 1.

1. Preventative Gynecology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Email:

Background: Ovarian epithelial tumors one of the most common gynecological neoplasms; we here evaluated the presence of HPV in benign and malignant examples. Methods: In this cross-sectional study the records of 105 patients with epithelial ovarian tumors (benign and malignant) referred to Imam Hossein University Hospital from 2012 to 2015 were evaluated along with assessment of the presence of the HPV infection using PCR. Results: Among 105 patients, comprising 26 (24.8%) with malignant and 79 (75.2%) with benign lesions, the factors found to impact on malignancy were age at diagnosis, age at first pregnancy, number of pregnancies and hormonal status. However, malignancies was not related to abortion, late menopause, and early menarche. In none of the ovarian tissues (benign and malignant) was HPV DNA found. Conclusion: In this study HPV DNA could not be found in any epithelial ovarian tumors (benign and malignant) removed from 105 women; more studies with larger sample size are needed for a definite conclusion.
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http://dx.doi.org/10.22034/APJCP.2017.18.5.1233DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5555528PMC
May 2017

Association of Mycobacterium infections in patients with Mendelian susceptibility to mycobacterial disease with venous thromboembolism.

Microbiol Immunol 2016 Oct;60(10):678-686

Chronic Respiratory Diseases Research Center, National Research Institute of Tuberculosis and Lung Diseases.

An association between a hypercoagulable state and Mendelian susceptibility to mycobacterial disease (MSMD) has been established in a few studies; resultant thrombosis is considered rare. In a case-control study, the prevalence of factor V Leiden, prothrombin G20210A and methylenetetrahydrofolate reductase (MTHFR) C677T, A1298C mutations were investigated in mycobacterium-infected patients. The study comprised 30 patients with mycobacterial infections (invasive, disseminated and/or recurrent infections with Bacille Calmette-Guerin or non-tuberculosis mycobacteria and Mycobacterium Tuberculosis with positive results for acid-fast bacilli and tuberculin skin tests) and 30 normal healthy controls. Forty female (66.7%) and 20 male subjects (33.3%) aged from 3 to 70 years were recruited into this study. Genotyping of targeted genes was performed by RT-PCR and cytokine TNF-α concentrations were quantified using a commercially available ELISA kit. Significant associations between mycobacterial infection and TNF-α production after stimulating peripheral blood mononuclear cells with LPS alone and with IFN-γ plus LPS were identified. Moreover, genotyping analysis in the studied population revealed a significant association between MTHFR c.677C>T (OR, 3.28; 95% CI, 1.35-7.92; P < 0.05), MTHFR c.1298A>C (OR, 2.33; 95% CI, 1.10-4.93; P < 0.05) and mycobacterial infection in affected patients, indicating susceptibility to venous thromboembolism according to previous studies. Additionally, mycobacterium-infected patients had a significantly greater prevalence of MTHFR C677T and A1298C mutations than controls.
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http://dx.doi.org/10.1111/1348-0421.12442DOI Listing
October 2016

Prevalence of HLA-B*5701 and Its Relationship with Abacavir Hypersensitivity Reaction in Iranian HIV-Infected Patients.

Tanaffos 2016 ;15(1):48-52

Virology Research Center, NRITLD, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Background: Hypersensitivity reaction (HSR) is a major adverse effect of abacavir (ABC), which occurs in 5-8% of Caucasians. The relationship between Human Leukocyte Antigen (HLA) and ABC HSR has been reported in various populations. It has been proposed to administer ABC only to HLA-B*5701 negative patients to avoid this reaction. The purpose of this study was to assess the prevalence of HLA-B*5701 in Iranian HIV positive patients. We also sought to find the relationship between this allele with ABC HSR in patients who received the medication.

Materials And Methods: We screened patients for HLA-B*5701 allele using SybrGreen real time PCR-melting method on blood samples from HIV positive patients who were referred to our hospital. The quality of the extracted genome was evaluated by B-globin housekeeping gene as internal control prior to HLA-B*5701 allele screening.

Results: Of 198 HIV-infected patients, 6 (3.0%) had the HLA-B*5701 allele (95% CI, 1%-5%). Among the 28 patients who were given ABC, one individual had the HLA-B*5701 allele and experienced ABC HSR.

Conclusion: Prevalence of HLA-B*5701 in Iranian patients was lower than that in Caucasians but was comparable with that of other Middle Eastern populations. Screening for HLA-B*5701 before ABC administration as part of antiretroviral therapy may reduce the risk of HSR.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4937762PMC
July 2016

Cytomegalovirus and Langerhans Cell Histiocytosis: Is There a Link?

Iran J Pediatr 2016 Apr 27;26(2):e673. Epub 2016 Feb 27.

Department of Community Oral Health, Dental Research Center, Dentistry Research Institute, Tehran University of Medical Sciences, Tehran, IR Iran.

Background: Langerhans cell histiocytosis is a rare proliferative histiocytic disease of unknown etiology. Histologically, it is characterized by granuloma-like proliferation of Langerhans-type dendritic cells derived from bone marrow. Many investigators have suggested the possible role of viruses such as Epstein-Barr virus, human herpesvirus-6 (HHV-6), herpes simplex virus (HSV) types 1 and 2, and Cytomegalovirus in the pathogenesis of Langerhans cell histiocytosis.

Objectives: In this study, we have investigated the presence of Cytomegalovirus in Langerhans cell histiocytosis in Iranian children.

Patients And Methods: In this retrospective study, we have investigated the presence of Cytomegalovirus DNA expression, using paraffin-embedded tissue samples of 30 patients with Langerhans cell histiocytosis and 30 age and site-matched controls by qualitative Polymerase Chain Reaction (PCR) method.

Results: No significant difference in prevalence of Cytomegalovirus presence between patients and controls was found. Cytomegalovirus was found by qualitative PCR in only 2 (6.66%) out of 30 patients and in 1 (3.3%) of 30 control samples with a P value of 1 (1.00 > 0.05) using chi-square test with OR: 2.07; 95% CI of OR: 0.18 - 24.15.

Conclusions: Our findings do not support the hypothesis of a possible role for Cytomegalovirus in the pathogenesis of Langerhans cell histiocytosis.
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http://dx.doi.org/10.5812/ijp.673DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4904486PMC
April 2016

Epidemiology of Rotavirus-Norovirus Co-Infection and Determination of Norovirus Genogrouping among Children with Acute Gastroenteritis in Tehran, Iran.

Iran Biomed J 2016 Nov 3;20(5):280-6. Epub 2016 May 3.

Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

Background: Enteric viruses, particularly human rotavirus and norovirus, have been shown to replace bacteria and parasites, as the most common pathogens responsible for acute diarrhea. However, there are still few epidemiological data on the simultaneous occurrence of these viruses in Iran. In this regard, the aim of this study was to assess the useful epidemiological data on the gastroenteritis associated with rotavirus-norovirus mixed infection and to examine the prevalence of norovirus genogrouping among children aged less than five years old in Iran.

Methods: A total of 170 stool samples were collected from children under five years of age with the clinical signs and symptoms of acute gastroenteritis, from May 2013 to May 2014. For the detection of both rotavirus and norovirus, total RNA was extracted from all samples, followed by reverse transcription polymerase chain reaction (RT-PCR). For both detected rotaviruses and noroviruses, genogrouping was performed.

Results: Of 170 samples, 49 (28.8%) and 15 (8.8%) samples were found to be positive for rotavirus and norovirus infections by RT-PCR. Interestingly, 6 (3.5%) patients were positive for both infections. Among the 15 norovirus-positive patients, 13 (86.6%) and 2 (13.3%) belonged to genogroups GII and GI.

Conclusions: The norovirus genogroup GII and rotavirus lead to the serious infections in children with acute gastroenteritis. However, more well-designed studies are needed to further elucidate the role of other enteric viruses in acute gastroenteritis.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5075141PMC
http://dx.doi.org/10.22045/ibj.2016.05DOI Listing
November 2016

Detection of Epstein-Barr Virus DNA in Langerhans Cell Histiocytosis.

Jundishapur J Microbiol 2015 Dec 26;8(12):e27219. Epub 2015 Dec 26.

Department of Community Oral Health, Dental Research Center, Dentistry Research Institute, School of Dentistry, Tehran University of Medical Sciences, Tehran, IR Iran.

Background: Langerhans cell histiocytosis (LCH) is a rare histiocytic proliferation of unknown etiology. It is characterized by granuloma-like proliferation of Langerhans-type dendritic cells and mainly affects young children. Although multiple investigators have suggested the possible role of viruses, such as Epstein-Barr virus (EBV), human herpesvirus-6 (HHV-6), Herpes simplex virus (HSV) types 1 and 2, and Cytomegalovirus (CMV) in the pathogenesis of LCH, it remains, however, debated.

Objectives: The EBV infection is reported to be associated with LCH. Nevertheless, no report could be found about involved Iranian children in English medical literature. In this study, we investigated the presence of EBV in Iranian children with LCH.

Patients And Methods: In this retrospective study, in which we investigated the prevalence of presence of EBV DNA in LCH, using paraffin-embedded tissue samples of 30 patients with LCH and 30 age and tissue-matched controls, who were operated for reasons other than infectious diseases (between the years 2002 and 2012), by real-time polymerase chain reaction (RT-PCR) method, in the department of pediatric pathology. No ethical issues arose in the study, because only the pathology reports were reviewed, retrospectively, and the patients were anonymous.

Results: There was a significant difference in prevalence of EBV presence between patients and controls. The EBV was found by RT-PCR in 19 (63.33%) out of 30 patients and only in eight (26.7%) of 30 control samples. The P = 0.004, was calculated using chi-square test (OR: 4.75; 95% CI: 1.58 ‒ 14.25).

Conclusions: Our study is the first investigation performed on patients with LCH and its possible association with EBV in Iran. Considering the P = 0.004, which is statistically significant, the findings do support the hypothesis of a possible role for EBV in the pathogenesis of LCH. These results are in accordance with several previous investigations, with positive findings.
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http://dx.doi.org/10.5812/jjm.27219DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4746794PMC
December 2015

Telomere Shortening in Blood Leukocytes of Patients with Chronic Obstructive Pulmonary Disease.

Tanaffos 2015 ;14(1):10-6

Chronic Respiratory Diseases Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Introduction: Chronic obstructive pulmonary disease (COPD) is characterized by airflow limitation that is not completely reversible by administration of inhaled bronchodilators. Many studies propose that telomere length shortening might have occurred in COPD patients. We aimed to determine the telomere length in COPD patients and compare the results of non-smoking and smoking control subjects.

Materials And Methods: In our case-control study, 84 clinically stable COPD patients were recruited on admission to Masih Daneshvari Hospital. Eighty-five healthy controls were also selected including 45 non-smokers and 40 smokers admitted for diseases other than COPD. Spirometry was done for all subjects. Telomere length was measured by quantitative real time PCR as described by Cawthon. The telomere repeat copy number (T) to single-gene copy number (S) ratio was calculated using the comparative Ct method.

Results: The mean ±SD of age was 64.33±10.04 years in patients and 65.06 ±10.02 years in controls (P=0.693). The mean ±SD of FEV1 was 1.62±0.75 L in patients, 2.84±0.54 L in smoker controls and 2.83±0.56 L in non-smoker controls; significant differences were detected in this regard between cases and controls (P<0.001). T/S ratio was significantly lower in COPD patients (0.61±0.08) than in the control subjects (0.69±0.09) (P<0.001). However, telomere length was shorter in the patients than in controls in each age group (P<0.001). Additionally, there were no statistically significant differences in telomere length between the smoker and non-smoker control subjects. Regarding the correlation between BMI and telomere length, there were no significant differences among the patients and control groups.

Conclusion: In conclusion, we found that telomere length in COPD patients was shorter than that in smoker and non-smoker controls, irrespective of age, sex, spirometric variables, BMI and history of cigarette smoking.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4515325PMC
July 2015

Disseminated Kaposi's Sarcoma with the Involvement of Penis in the Setting of HIV Infection.

Indian J Dermatol 2015 Jan-Feb;60(1):104

Clinical Tuberculosis and Epidemiology Research Center, National Research Institute of Tuberculosis and Lung Disease, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Kaposi's sarcoma (KS) is a malignant proliferation of the endothelial cells. It typically presents with several vascular nodules on the skin and other organs. The penile localization of KS, particularly on the shaft area, is exceptional. We report an HIV-positive 34-year-old man who had multiple purplish-black plaques on his extremities and several small violaceous macules on the glans and shaft of the penis. Kaposi's sarcoma was diagnosed by histopathology.
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http://dx.doi.org/10.4103/0019-5154.147852DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4318030PMC
February 2015

Secondary infection and clinical aspects after pandemic swine-origin influenza a (H1N1) admission in an Iranian critical care unite.

Int J Crit Illn Inj Sci 2014 Oct-Dec;4(4):309-13

Mycobacteriology Research Center Virology Research Center, National Research Institute of Tuberculosis and Lung Disease, Masih Daneshvari Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Objective: A new flu virus (H1N1) swine origin and cause of human infection with acute lung disease was published in the world and led to many patients were admitted in intensive care unit (ICU).

Materials And Methods: In a prospective descriptive study, all ICU patients in a pulmonary disease specialist hospital between April 2010 and July 2011 with confirmed infection (H1N1) were evaluated. Information including demographic, clinical and microbiology using Statistical Package for Social Sciences (SPSS) software version 16 was studied and classified.

Results: Of 46 patients hospitalized with confirmed diagnosis of swine flu pneumonia (H1N1), 20 cases (43.7%) admitted in ICU out of which 10 cases were males (50%), the mean age was 36.9 and the range was 21-66 years. Nine patients (45%) had underlying diseases. Most underlying disease was respiratory disease in which four cases (20%) were of asthma and one patient had chronic obstructive pulmonary disease (COPD). No admission of pregnant patient with swine flu was reported in the ICU. Cough and sputum were the most frequent symptoms (19 patients equal 95%). Four patients (20%) were admitted with decreased level of consciousness and five cases (25%) died during hospitalization.

Conclusion: It seems, swine flu with high mortality and transfer rates is a worldwide health problem. Because of limited treatment regimen, the risk of secondary infection and high need to intensive care in H1N1 pneumonia, environmental control, including vaccination of high risk people and public announcement, make determining role in controlling of this disease.
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http://dx.doi.org/10.4103/2229-5151.147536DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4296334PMC
January 2015