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Chronic hepatitis C virus infection is a major cause of mortality due to liver cirrhosis globally. Despite the advances in recent therapeutic strategies, there is yet a high burden of HCV-related cirrhosis worldwide concerning low coverage of newly developed antiviral therapies, insufficient validity of the current diagnostic methods for cirrhosis, and incomplete understanding of the pathogenesis in this stage of liver disease. Hence we aimed to clarify the molecular events in HCV-related cirrhosis and identify a liver-specific gene signature to potentially improve diagnosis and prognosis of the disease.Through RNA-seq transcriptome profiling of liver samples of Iranian patients with HCV-related cirrhosis, the differentially expressed genes (DEGs) were identified and subjected to functional annotation including biological process (BP) and molecular function (MF) analysis and also KEGG pathway enrichment analysis. Furthermore, the validation of RNA-seq data was investigated for seven candidate genes using qRT-PCR. Moreover, the diagnostic and prognostic power of validated DEGs were analyzed in both forms of individual DEG and combined biomarkers through receiver operating characteristic (ROC) analysis. Finally, we explored the pair-wise correlation of these six validated DEGs in a new approach.We identified 838 significant DEGs (padj ˂0.05) enriching 375 and 15 significant terms subjected to BP and MF, respectively (false discovery rate ˂ 0.01) and 46 significant pathways (p-value ˂ 0.05). Most of these biological processes and pathways were related to inflammation, immune responses, and cellular processes participating somewhat in the pathogenesis of liver disease. Interestingly, some neurological-associated genes and pathways were involved in HCV cirrhosis-related neuropsychiatric disorders. Out of seven candidate genes, six DEGs, including inflammation-related genes ISLR, LTB, ZAP70, KLRB1, and neuronal-related genes MOXD1 and Slitrk3 were significantly confirmed by qRT-PCR. There was a close agreement in the expression change results between RNA-seq and qRT-PCR for our candidate genes except for SAA2-SAA4 (P= 0.8). High validity and reproducibility of six novel DEGs as diagnostic and prognostic biomarkers were observed. We also found several pair-wise correlations between validated DEGs.Our findings indicate that the six genes LTB, ZAP70, KLRB1, ISLR, MOXD1, and Slitrk3 could stand as promising biomarkers for diagnosing of HCV-related cirrhosis. However, further studies are recommended to validate the diagnostic potential of these biomarkers and evaluate their capability as targets for the prevention and treatment of cirrhosis disease.

Abstract Background The antiviral properties of metal nanoparticles against various viruses, including those resistant to drugs, are currently a subject of intensive research. Recently, the green synthesis of nanoparticles and their anti-viral function have attracted a lot of attention. Previous studies have shown promising results in the use of Arabic gum for the green synthesis of nanoparticles with strong antiviral properties. In this study we aimed to investigate the antiviral effects of MnO2 nanoparticles (MnO2-NPs) synthesized using Arabic gum, particularly against the influenza virus. Methods Arabic gum was used as a natural polymer to extract and synthesize MnO2-NPs using a green chemistry approach. The synthesized MnO2-NPs were characterized using SEM and TEM. To evaluate virus titration, cytotoxicity, and antiviral activity, TCID50, MTT, and Hemagglutination assay (HA) were performed, respectively. Molecular docking studies were also performed to investigate the potential antiviral activity of the synthesized MnO2-NPs against the influenza virus. The molecular docking was carried out using AutoDock Vina software followed by an analysis with VMD software to investigate the interaction between Arabic gum and the hemagglutinin protein. Results Simultaneous combination treatment with the green-synthesized MnO2-NPs resulted in a 3.5 log HA decrement and 69.7% cellular protection, which demonstrated the most significant difference in cellular protection compared to the virus control group (p-value

Abstract Background Human papillomavirus (HPV) is one of the most prevalent sexually transmitted diseases worldwide. The present review was conducted to accumulate evidence on the relationship between cervicovaginal human papillomavirus infection and serum vitamin D status. Methods Electronic databases including Web of Science, Embase, Scopus, and PubMed were searched by different combinations of keywords related to “human papillomavirus” and “vitamin D”, obtained from Mesh and Emtree with AND, and OR operators without any time restriction until December 24, 2022. Selection of articles was based on the inclusion and exclusion criteria. Newcastle-Ottawa Scale was used for quality assessment. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) checklist was applied for reporting. Results In total, 276 citations were retrieved. After removing duplicates, and non-related articles, the full texts of 7 articles were reviewed including 11168 participants. Three studies reported that there was a positive relationship between vitamin D deficiency and cervicovaginal human papillomavirus while three studies did not. One study showed a significant positive association between higher vitamin D stores and short-term high-risk human papillomavirus persistence. Conclusions The findings showed no firm evidence for any association between serum vitamin D level and cervicovaginal human papillomavirus infection, although the possible association could not be discarded. Further investigations are needed to reach sound evidence.

Abstract Malaria has not yet been eradicated in Iran, and Plasmodium vivax (P. vivax) is the main cause of malaria in the country. This study aimed to investigate and analyze the amount of genetic diversity of Plasmodium vivax merozoite surface protein-5 (PvMSP-5) exon 1 gene in the southeast of Iran. Thirty-five patients with clinical symptoms of P. vivax malaria participated. The exon 1 of PvMSP-5 was amplified by PCR, and the PCR product of all isolates was sequenced, and genetic polymorphisms were determined using various genetic software. The analysis showed that studied isolates are different from one another in the DnaSP software version. Out of the 612 sites, 477 were monomorphic and 135 were segregated. The total number of mutations was 143. The singleton variable and the parsimony informative sites were 23 and 112, respectively. There were 17 specific haplotypes with haplotype diversity equal to 0.943. Nucleotide diversity was equal to 0.06766 in the isolates. The ratio of nonsynonymous (0.06446) to synonymous (0.07909) mutations was 0.815020. Tajima’s D, which expressed coding, and non-coding regions, was 0.72403, which was not deemed significant (P > 0.10). The analysis of intrapopulation diversity revealed nucleotide and haplotype diversity in the msp-5 gene of Iranian P. vivax isolates. In addition to balancing or purifying selection, intragenic recombination also contributed to the variation observed in exon 1 of PvMSP-5, according to the findings.

Background: Toxoplasma gondii infects nearly one-third of the world's population. Due to the significant side effects of current treatment options, identifying safe and effective therapies seems crucial. Nanoparticles (NPs) are new promising compounds in treating pathogenic organisms. Currently, no research has investigated the effects of zinc oxide NPs (ZnO-NPs) on Toxoplasma parasite. We aimed to investigate the therapeutic efficacy of ZnO-NPs against tachyzoite forms of T. gondii, RH strain in BALB/c mice. Methods: In an experiment with 35 female BALB/c mice infected with T. gondii tachyzoites, colloidal ZnO-NPs at concentrations of 10, 20, and 50 ppm, as well as a 50 ppm ZnO solution and a control group, were orally administered four hours after inoculation and continued daily until the mices’ death. Survival rates were calculated and tachyzoite counts were evaluated in the peritoneal fluids of infected mice. Results: The administration of ZnO-NPs resulted in the reduction of tachyzoite counts in infected mice compared to both the ZnO-treated and control group (P

Background: Cutaneous leishmaniasis (CL) is a parasitic disease that presents a broad spectrum of clinical features. Treatment of CL is problematic. We aimed to compare the field therapeutic efficacy of topical nanoliposomes containing 0.4% amphotericin B (Nano Lip-AmB) alone and in combination with cryotherapy and/or Glucantime® on human CL in the endemic areas of Iran. Methods: This retrospective study was performed based on the results of using Nano Lip-AmB alone or with Glucantime® and/or cryotherapy in the treatment of zoonotic cutaneous leishmaniasis (ZCL) in patients referred to health centers of Isfahan, Golestan and Ilam Provinces of Iran as endemic foci of ZCL caused by Leishmania major besides Mashhad and Bam cities as endemic foci of anthroponotic cutaneous leishmaniasis (ACL) caused by with L. tropica. Results: Two hundred and seventy-eight patients with CL were included in the current study. All of the patients (100%) who received Nano Lip-AmB alone or in combination with Glucantime® and/or cryotherapy based on guideline of Iranian national committee for the treatment of CL. Two patients with 7 skin lesions, who was resident in ACL endemic area and received Nano Lip-AmB plus Glucantime® and another patient was a resident of ZCL endemic area and received Nano Lip-AmB plus cryotherapy showed clinical relapses after treatment. Conclusion: Sina Ampholeish® in combination with other standard protocols of treatment of CL is well tolerated and with acceptable clinical efficacy rate.

Background: Visceral leishmaniasis (VL) is one of the most important neglected tropical diseases. The zoonotic form of VL is endemic in some areas of Iran. We aimed to determine the status of VL identified in humans and canines in different parts of Iran from 2013 to 2022. Method: A national representative cross-sectional study was conducted in 10 provinces of Iran, including the national leishmaniasis reference lab. We employed the direct agglutination test (DAT) as a reliable serological method to detect anti-Leishmania infantum antibodies in humans and animal reservoir hosts. Additionally, a narrative literature review was conducted to identify relevant studies on VL seroprevalence in Iran from 2013 to 2023. Results: The results of 21281 human and 5610 canine serum samples from 2013 to 2022 are reported. Altogether, 448 (2.1%, 95%CI: 2.0-2.3) human serum samples showed anti-L. infantum antibody levels of ≥1:3200. Of these samples, 13716 (64.5%) were collected actively, which showed a seroprevalence of 0.6% (95% CI: 0.5-0.8) and 7565 (35.5%) were collected passively, which showed a seroprevalence of 4.8% (95%CI: 4.3-5.3). Overall, 1035 (20.1%, 95%CI: 19.0-21.2) of 5160 domestic dogs (Canis familiaris) samples showed anti-L. infantum antibody levels of ≥1:320. Northwest (2.8%) and northeast (0.96%) regions had the highest human VL seroprevalence, while northwest (21.5%) and south (14.4%) regions had the highest canine VL seroprevalence. Conclusion: Zoonotic VL, an endemic parasitic disease, is still present in several different distinct areas across Iran. While human VL cases have shown a declining trend over the last decade, the prevalence of canine VL remains significant.

Background: Cutaneous leishmaniasis (CL) is an endemic infection in the Middle East, including Iran that is also spreading to new foci. We aimed to determine the leishmaniasis species causing CL in Alborz province. Methods: Overall, out of 55-suspected CL patients referred to health centers in Alborz Province, north central Iran in 2019, 40 patients had positive smear for CL based on optical microscopy. The internal transcribed spacer 1 (ITS1) of nuclear ribosomal DNA (rDNA) was amplified by PCR. Leishmania species were identified by PCR–restriction fragment length polymorphism (PCR-RFLP) using BshF I (Hae III) enzyme. Results: Out of the 40 positive patients with CL, 34 cases (85%) had been caused by Leishmania (L) major and six (15%) by L. tropica. Fifteen patients had no history of traveling to the disease endemic areas, of which nine were Iranians. Skin lesions and scars caused by CL were mostly observed on the hands and face. Moreover, more than two skin lesions were observed in 22 cases (55%), all of which were infected with L. major. A single skin ulcer was seen in 18 (45%) of the CL patients. Conclusion: Climate change, reduced rainfall, and demographic changes such as migration into Alborz Province and the increasing marginalization of the population and their entry to settle in new areas might have caused natural transmission of both L. tropica and L. major in this province.

Background: Currently, there are conflicting reports on the associations between Toxoplasma gondii infection and multiple sclerosis (MS) in humans. In the present study, a case–control study was carried out to assess associations between seropositivity to T. gondii infection and MS. Methods: This case-control study was carried out on 200 MS patients (cases) attended in Sina Hospital affiliated to Tehran University of Medical Sciences, Tehran, Iran, and 200 healthy subjects from the general population of the same city, March to July 2017. Blood samples were collected from individuals and were examined using Enzyme-linked immunosorbent assay (ELISA) for the presence of T. gondii IgG antibodies and the IgG-positive samples were further analyzed for specific anti-T. gondii IgM. Results: The overall seroprevalence of anti-T. gondii IgG was 44.2% (177/400) in 121 (60.5%) sera of the 200 MS patients (cases) and 56 (28.0%) sera of the 200 controls (OR = 3.94; 95% CI: 2.59–5.99; P < 0.001). Seroprevalence of T. gondii infection in MS patients increased significantly with increasing of age (P < 0.001). In the control group, no statistically significant differences were seen between the seroprevalence of T. gondii infection in various age groups (P = 0.858). Moreover, no statistically significant relationships were reported between the seropositivity to T. gondii and the sex for the cases and controls (P>0.05). Anti-T. gondii IgM antibodies were not detected in anti-T. gondii IgG positive patients. Conclusion: T. gondii infection might be a probability risk factor for MS. However, further studies are necessary to describe clearly the roles of T. gondii infection in MS.

Abstract Background MDA-7/IL-24 cytokine has shown potent antitumor properties in various types of cancer without exerting any significant toxicity on healthy cells. It has also been proved to encompass pro-immune Th1 cytokine-like behavior. Several E7 DNA vaccines have developed against human papillomavirus (HPV)-related cervical cancer. However, the restricted immunogenicity has limited their clinical applications individually. To address this deficiency, we investigated whether combining the E7 DNA vaccine with MDA-7/IL-24 as an adjuvant would elicit efficient antitumor responses in tumor-bearing mouse models. Next, we evaluated how suppression of immunosuppressive IL-10 cytokine would enhance the outcome of our candidate adjuvant vaccine. Methods For this purpose, tumor-bearing mice received either E7 DNA vaccine, MDA-7/IL-24 cytokine or combination of E7 vaccine with MDA-7/IL-24 adjuvant one week after tumor challenge and boosted two times with one-week interval. IL-10 blockade was performed by injection of anti-IL-10 mAb before each immunization. One week after the last immunization, mice were sacrificed and the treatment efficacy was evaluated through immunological and immunohistochemical analysis. Moreover, the condition of tumors was monitored every two days for six weeks intervals from week 2 on, and the tumor volume was measured and compared within different groups. Results A highly significant synergistic relationship was observed between the E7 DNA vaccine and the MDA-7/IL-24 cytokine against HPV-16+ cervical cancer models. An increase in proliferation of lymphocytes, cytotoxicity of CD8+ T cells, the level of Th1 cytokines (IFN-γ, TNF-α) and IL-4, the level of apoptotic markers (TRAIL and caspase-9), and a decrease in the level of immunosuppressive IL-10 cytokine, together with the control of tumor growth and the induction of tumor regression, all prove the efficacy of adjuvant E7&IL-24 vaccine when compared to their individual administration. Surprisingly, vaccination with the DNA E7&IL-24 significantly reduced the population of Regulatory T cells (Treg) in the spleen of immunized mice compared to sole administration and control groups. Moreover, IL-10 blockade enhanced the effect of the co-administration by eliciting higher levels of IFN-γ and caspase-9, reducing Il-10 secretion and provoking the regression of tumor size. Conclusion The synergy between the E7 DNA vaccine and MDA-7/IL-24 suggests that DNA vaccines’ low immunogenicity can be effectively addressed by coupling them with an immunoregulatory agent. Moreover, IL-10 blockade can be considered a complementary treatment to improve the outcome of conventional or novel cancer therapies.

Abstract Background Ocular infection with Toxoplasma gondii is a major preventable cause of blindness, especially in young people. The aim of the present study was to assess detection rate of T. gondii DNA in blood samples of clinically diagnosed of ocular toxoplasmosis using uracil DNA glycosylase-supplemented loop-mediated isothermal amplification (UDG-LAMP) and real-time quantitative PCR (qPCR) based on REP-529 and B1. Methods One hundred and seventeen patients with clinically diagnosed ocular toxoplasmosis (OT) were participated in the study as well as 200 control patients. Peripheral blood samples were assessed using UDG-LAMP and qPCR techniques targeting REP-529 and B1. Results Detection limits of qPCR using REP-529 and B1 were estimated as 0.1 and 1 fg of T. gondii genomic DNA, respectively. The limits of detection for UDG-LAMP using REP-529 and B1 were 1 and 100 fg, respectively. In this study, 18 and 16 patients were positive in qPCR using REP-529 and B1, respectively. Based on the results of UDG-LAMP, 15 and 14 patients were positive using REP-529 and B1, respectively. Results of the study on patients with active ocular lesion showed that sensitivity of REP-529 and BI targets included 64 and 63%, respectively using qPCR. Sensitivity of 62 and 61%, were concluded from UDG-LAMP using REP-529 and B1 in the blood cases of active ocular lesion. qPCR was more sensitive than UDG-LAMP for the detection of Toxoplasma gondii DNA in peripheral blood samples of patients with clinically diagnosed toxoplasmic chorioretinitis. Furthermore, the REP-529 included a better detection rate for the diagnosis of ocular toxoplasmosis in blood samples, compared to that the B1 gene did. Moreover, the qPCR and UDG-LAMP specificity assessments have demonstrated no amplifications of DNAs extracted from other microorganisms based on REP-529 and B1. Conclusions Data from the current study suggest that qPCR and UDG-LAMP based on the REP-529 are promising diagnostic methods for the diagnosis of ocular toxoplasmosis in blood samples of patients with active chorioretinal lesions.

Abstract Background Despite recent advancements, limitations in the treatment and control of hepatitis C virus (HCV) infection reprioritized the studies for invention of an efficient HCV vaccine to elicit strong neutralizing antibodies (NAbs) and cellular responses. Methods Herein, we report molecular construction of a BacMam virus-based surface display for a subtype-1a HCV gpE2 (Bac-CMV-E2-gp64; Bac) that both expressed and displayed gpE2 in mammalian cells and bacouloviral envelope, respectively. Results Assessments by western blotting, Immunofluorescence and Immunogold-electron microscopy indicated the proper expression and incorporation in insect cell and baculovirus envelope, respectively. Mice immunized in three different prime-boost immunization groups of: Bac/Bac, Bac/Pro (bacoulovirus-derived gpE2) and Bac/DNA (plasmid DNA (pCDNA)-encoding gpE2) developed high levels of IgG and IFN-γ (highest for Bac/Bac group) indicating the induction of both humeral and cellular immune responses. Calculation of the IgG2a/IgG1 and IFN-γ/IL-4 ratios indicated a Th1 polarization of immune responses in the Bac/Bac and Bac/DNA groups but a balanced Th1-Th2 phenotype in the Bac/Pro group. Sera of the mice in the Bac/Bac group provided the highest percentage of cross-NAbs against a subtype-2a HCVcc (JFH1) compared to Bac/Pro and Bac/DNA groups (62% versus 41% and 6%). Conclusions Results indicated that BacMam virus-based surface display for gpE2 might act as both subunit and DNA vaccine and offers a promising strategy for development of HCV vaccine for concurrent induction of strong humoral and cellular immune responses.

Background: Toxoplasma gondii infection (toxoplasmosis) has the potential to cause a serious disease in immunocompromised patients and can be fatal in this population. We conducted a systematic review and meta-analysis to assess comprehensively the pooled seroprevalence of toxoplasmosis among immunocompromised patients including HIV/AIDS patients, cancer patients, and transplant recipients in Iran. Methods: PubMed, Web of Science, Scopus, Embase, and Google Scholar databases (international) and Scientific Information Database (SID), Magiran, IranMedex, and IranDoc databases (national) were systematically searched for all reports that possibly contained data for T. gondii prevalence in different immunocompromised populations in Iran between 2013 and 2022. Results: Overall, IgG seroprevalence rate of toxoplasmosis in Iranian immunocompromised patients was 45.1% (95% confidence interval (CI), 37.4–52.9). IgG seroprevalence rate of toxoplasmosis in 12 studies that included 2279 cancer patients, 19 studies that included 2565 HIV/AIDS patients and in 3 studies that included 200 transplant recipients was 43.6% (95% CI, 30.2–57.0), 45.9% (95% CI, 34.8–57.1) and 45.8% (95% CI, 32.5–59.0), respectively. Moreover, IgM seroprevalence rate in the 26 studies was 2.6% (95% CI, 1.4–3.7). Conclusion: Our findings represent a high seroprevalence rate of Toxoplasma IgG among immunocompromised patients. Health improvement and education toward prevention of toxoplasmosis is of great importance for these susceptible populations.

Background: Toxoplasma gondii is a neuroinvasive protozoa pathogen that could manipulate its intermediate host's behavior. However, the possible link between T. gondii infection and the development of neurodegenerative disorders such as Parkinson’s disease (PD) has been proposed, we tested the hypothesis that in chronic toxoplasmosis neuroinflammation, and molecular mediators potentiate behavioral-cognitive impairments in BALB/c mice with PD. Methods: To establish chronic toxoplasmosis by Tehran strain, cysts of T. gondii were injected intraperitoneally into BALB/c mice in Kerman, Iran in 2019. To induce the PD model, mice (BALB/c) were treated with Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The behavioral experiments such as anxiety and motor coordination were performed using the Open field and Rotarod tests. Additionally, we investigated the contribution of Toxoplasma-induced neuroinflammation, and behavioral-cognitive impairments in the PD mice model. Results: Chronic toxoplasmosis caused PD-like symptoms and induced various behavioral changes in infected BALB/c mice. In T. gondii infected+MPTP treated group, T. gondii infection could potentiate PD in infected mice receiving MPTP and caused remarkable dysfunction in motor coordination and change in anxiety and depression-like behaviors similar or more severe than PD group. Conclusion: Chronic T. gondii infection exacerbates pathological progression of PD in BALB/c mice brain by promoting neuroinflammation, and behavioral changes establishing.

Chronic granulomatous disease (CGD) described as an essential immunodeficiency problem of phagocytic cells resulting in a phagocyte dysfunction and inability to kill a spectrum of bacteria and fungi. Despite the fact that CGD patients are more susceptible to intracellular infections, visceral leishmaniasis has been reported rarely in these cases. Here, we report an uncommon case of visceral leishmaniasis in a child with CGD. An 8-yr old boy with CGD presented to the infectious disease ward, Children's Medical Center, Tehran University of Medical Sciences, Iran after the onset of 20 days fever with chronic crusted ulcer approximately 3 cm × 3cm on the left upper limb and a small ulcer measuring 0.5 cm × 0.5 cm on the right knee with moderate secretion. Bone Marrow Aspiration (BMA) and Bone Marrow Biopsies (BMB) of fragmented samples were performed and polymorphic population of hematopoietic cells, Megakaryocytes and Leishman bodies were seen. The patient was treated with meglumine antimoniate (Glucantime®) 20 mg/kg for 28 days and after partial improvement patient discharged and continue the treatment at home. Amphotericin B lipid complex (Ambisome®) (3–5 mg/kg per dose once) was administered every 3 -4 weeks for 18 months as secondary prophylaxis that was well tolerated and effective.

Abstract Background Direct agglutination test (DAT) as a simple, accurate and reliable method, has been widely used for serodiagnosis of visceral leishmaniasis (VL) during the last three decades. The present study is a systematic review and meta-analysis to evaluate the diagnostic accuracy of DAT for serodiagnosis of human VL. Methods Electronic databases, including MEDLINE (via PubMed), SCOPUS, Web of Science, SID and Mag Iran (two Persian scientific search engines) were searched from December 2004 to April 2019. We determined the pooled sensitivity and specificity rates of DAT for the diagnosis of human VL, calculated positive and negative likelihood ratios (LR+ and LR-), and constructed summary receiver operating characteristic (ROC) curves parameters across the eligible studies. Results Of the 2928 records identified in the mentioned electronic databases and after examining reference lists of articles, 24 articles met inclusion criteria and were enrolled in the systematic review and out of them 20 records qualified for meta-analysis. The pooled sensitivity and specificity rates of DAT was 96% [95% CI, 92–98] and 95% [CI95% 86–99], respectively. The likelihood ratio of a positive test (LR+) was found to be 21 [CI95%, 6.6–66.5] and the likelihood ratio of a negative test (LR−) was found to be 0.04 [(CI95%, 0.02–0.08]. The combined estimate of the diagnostic odds ratio for DAT was high [467 (CI95%, 114–1912]). We found that the summary receiver operating characteristic curve (SROC) is positioned near the upper left corner of the curve and the area under curve (AUC) was 0.98 (95% CI, 0.97 to 0.99). Conclusion Referring to our analysis, we determined that DAT can be considered as a valuable tool for the serodiagnosis of human VL with high sensitivity and specificity. As DAT is a simple, accurate and efficient serological test, it can be recommended for serodiagnosis of human VL particularly in endemic areas.

Abstract Background Influenza virus infection is among the most detrimental threats to the health of humans and some animals, infecting millions of people annually all around the world and in many thousands of cases giving rise to pneumonia and death. All those health crises happen despite previous and recent developments in anti-influenza vaccination, suggesting the need for employing more sophisticated methods to control this malign infection. Main body The innate immunity modules are at the forefront of combating against influenza infection in the respiratory tract, among which, innate T cells, particularly gamma-delta (γδ) T cells, play a critical role in filling the gap needed for adaptive immune cells maturation, linking the innate and adaptive immunity together. Upon infection with influenza virus, production of cytokines and chemokines including CCL3, CCL4, and CCL5 from respiratory epithelium recruits γδ T cells at the site of infection in a CCR5 receptor-dependent fashion. Next, γδ T cells become activated in response to influenza virus infection and produce large amounts of proinflammatory cytokines, especially IL-17A. Regardless of γδ T cells’ roles in triggering the adaptive arm of the immune system, they also protect the respiratory epithelium by cytolytic and non-cytolytic antiviral mechanisms, as well as by enhancing neutrophils and natural killer cells recruitment to the infection site. Conclusion In this review, we explored varied strategies of γδ T cells in defense to influenza virus infection and how they can potentially provide balanced protective immune responses against infected cells. The results may provide a potential window for the incorporation of intact or engineered γδ T cells for developing novel antiviral approaches or for immunotherapeutic purposes.

Abstract Background The spread of airborne infectious diseases such as measles is a critical public health concern. The U.S. was certified measles-free in 2000, but the number of measles cases has increased in recent years breaking the record of the nationwide annual number of cases since 1992. Although the characteristics of schools have made them one of the most vulnerable environments during infection outbreaks, the transmission risk of measles among students is not completely understood. We aimed to evaluate how three factors influence measles transmission in schools: personal (vaccination), social (compartmentalizing), and building systems (ventilation, purification, and filtration). Methods We used a combination of a newly developed multi-zone transient Wells-Riley approach, a nationwide representative School Building Archetype (SBA) model, and a Monte-Carlo simulation to estimate measles risk among U.S. students. We compared our risk results with the range of reported transmission rates of measles in school outbreaks to validate the risk model. We also investigated the effectiveness of vaccination and ten supplemental infection control scenarios for reducing the risk of measles transmission among students. Results Our best nationwide estimate of measles transmission risk in U.S. schools were 3.5 and 32% among all (both unvaccinated and immunized) and unvaccinated students, respectively. The results showed the transmission risk of measles among unvaccinated students is > 70 times higher than properly immunized ones. We also demonstrated that the transmission risk of measles in primary schools (assuming teacher self-contained classrooms) is less than secondary schools (assuming departmentalized systems). For building-level interventions, schools with ductless-with-air-filter and ductless-without-air-filter systems have the lowest and highest transmission risks of measles, respectively. Finally, our simulation showed that infection control strategies could cut the average number of infected cases among all students in half when a combination of advanced air filtration, ventilation, and purification was adopted in the modeled schools. Conclusions Our results highlight the primary importance of vaccination for reducing the risk of measles transmission among students. Yet, additional and significant risk reduction can be achieved through compartmentalizing students and enhancing building ventilation and filtration systems.

Abstract Background Human papillomavirus (HPV)-associated malignancy remain a main cause of cancer in men and women. Cancer immunotherapy has represented great potential as a new promising cancer therapeutic approach. Here, we report Mesenchymal stem cells (MSCs) as a carrier for the delivery of oncolytic Newcastle disease virus (NDV) for the treatment of HPV-associated tumor. Methods For this purpose, MSCs obtained from the bone marrow of C57BL mice, then cultured and characterized subsequently by the flow cytometry analysis for the presence of cell surface markers. In this study, we sought out to determine the impacts of MSCs loaded with oncolytic NDV on splenic T cell and cytokine immune responses, caspase-3 and -9 expression, and myeloid and myeloid-derived suppressor cells (MDSCs) by histological and immunohistochemical studies in the tumor microenvironment (TME). Results Our findings proved that MSCs possess both migratory capacity and tumor tropism toward transplanted tumor tissue after peritumoral administration. Tumor therapy experiments indicated that oncolytic NDV delivered by MSCs-engineered system significantly reduces tumor growth, which is associated with the enhancement of E7-specific lymphocyte proliferation, CD8+ T cell cytolysis responses, and splenic IFN-γ, IL-4 and IL-12 responses compared with control groups. Moreover, the treatment upregulated the concentration of apoptotic proteins (caspase 9) and increased infiltration of tumor microenvironment with CD11b + myeloid and Gr1 + MDSCs cells. Conclusions Our data suggest MSCs carrying oncolytic NDV as a potentially effective strategy for cancer immunotherapy through inducing splenic Th1 immune responses and apoptosis in the tumor microenvironment.

Abstract Background Echinococcosis is a zoonotic disease caused by the larval stages of taeniid cestodes of the genus Echinococcus. The two major types of infection in humans are cystic echinococcosis (CE) or hydatidosis and alveolar echinococcosis (AE). It is endemic in some parts of the world, such as the Middle East, with Iran being a part of it. This systematic review and meta-analysis were conducted to determine the prevalence of CE and AE echinococcosis and their epidemiological and clinical aspects in Iran. Methods Electronic databases, including MEDLINE (via PubMed), SCOPUS, Web of Science, SID and Mag Iran (two Persian scientific search engines) were searched from 1 January 1990 to 8 August 2017. The prevalence of CE and AE echinococcosis was estimated using the random effects meta-analysis. Heterogeneity was evaluated by subgroup analysis. Data were analyzed by STATA version 12. Results Of the 2051 records identified in the mentioned electronic databases, Seventy-eight articles met our eligibility criteria, with a total of 214124individuals. The meta-analysis was performed on only 37 out of 78 included studies. The pooled prevalence of CE and AE in Iran was 5% [95% confidence interval )CI(: 3-6%] and 2% [95% CI: 0-5%], respectively. Subgroup meta-analysis revealed that the prevalence of CE was significantly higher in North [9%, 95% CI: 4-18%] and West of Iran [6%, 95% CI: 3-11%], patients younger than 40 years of age [7%, 95% CI: 4-12%], villagers and nomads [6%, 95% CI: 2-12%], and studies that used the combination of serological, clinical, and imaging diagnostic methods [7%, 95% CI: 5-9%]. There were no significant differences between the prevalence of CE among low and high-quality studies. Housewives were the most affected group by hydatidosis (n=24/77, 31%), followed by illiterate people (n=11/77, 14%) and farmers (n= 9/77, 12%). Liver [55%, 95% CI: 46-65%] and lung [28%, 95% CI, 22-35%] were the most common sites of cyst formation. Conclusions Given to the importance of echinococcosis on human health and domestic animals industry, it is necessary to implement monitoring and control measures in this regard.

Aref Teimouri,1,2 Mohammad Hossein Modarressi,3 Saeedeh Shojaee,1 Mehdi Mohebali,1,4 Mostafa Rezaian,1 Hossein Keshavarz1,41Department of Medical Parasitology and Mycology, Tehran University of Medical Sciences, Tehran, Iran; 2Students Scientific Research Center, Tehran University of Medical Sciences, Tehran, Iran; 3Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran; 4Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, IranCorrespondence: Hossein KeshavarzDepartment of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, PO Box 1417613191, Pour Sina Street, Ghods Avenue, Enghelab Avenue, Tehran, IranTel +98 218 895 1392Fax +98 218 895 1392Email hkeshavarz@tums.ac.irBackground: The aim of the present study was to develop a simple, portable, and rapid assay for serodiagnosis of toxoplasmosis based on recombinant Toxoplasma gondii (T. gondii) SAG1 (rSAG1) and GRA7 (rGRA7) proteins.Methods: The rSAG1 and rGRA7 proteins were expressed in Escherichia coli (E. coli) and purified in a single step by immobilized metal ion affinity chromatography. The immunoreactivity of the recombinant antigens was tested in an in-house IgG and IgM Dot enzyme-linked immunosorbent assay (Dot-ELISA) for potential use in serodiagnosis of T. gondii infection.Results: Results from the comparison of in-house rSAG1-Dot-ELISA with ELISA for the detection of anti-Toxoplasma IgG and IgM include sensitivity of 83.7% and 81.2%, specificity of 90.2% and 89.3%, positive predictive values of 85.9% and 68.4%, and negative predictive values of 88.6% and 94.3%, respectively. Sensitivity of 66.2%, specificity of 81.2%, positive predictive values of 71.6%, and negative predictive values of 77.1% were concluded from in-house IgG rGRA7-Dot-ELISA. The sensitivity and specificity of IgM rGRA7-Dot-ELISA included 87.5% and 83.9%, respectively. Sensitivity and specificity of in-house Dot-ELISA for a combination of rSAG1 and rGRA7 included 87.5% and 91.1% for IgG and IgM, respectively. Sensitivity and specificity of a combination of rSAG1 and rGRA7 for the detection of IgM in suspected sera to acute toxoplasmosis were higher than those for the detection of IgG in sera with chronic infections (90.6% and 92% instead of 86.2% and 91.6%, respectively).Conclusion: The highlighted parameters of combined recombinant proteins were more significant than those of single recombinant proteins in in-house Dot-ELISA. These data suggest that the in-house Dot-ELISA based on rSAG1 and rGRA7 combination is a promising diagnostic tool with a similar sensitivity to the native antigens of T. gondii, which can be used for the serodiagnosis of toxoplasmosis in fields as well as less equipped laboratories.Keywords: Toxoplasma gondii RH strain, in-house Dot-ELISA, rSAG1, rGRA7, soluble tachyzoite antigen (STAg), recombinant proteins

Sara Ayazian Mavi,1 Mohammad Hossein Modarressi,2 Mehdi Mohebali,1,3 Saeedeh Shojaee,1 Hojjat Zeraati,4 Aref Teimouri,1,5 Hossein Keshavarz1,31Department of Medical Parasitology and Mycology, Tehran University of Medical Sciences, Tehran, Iran; 2Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran; 3Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran; 4Department of Epidemiology and Biostatistics, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran; 5Students Scientific Research Center, Tehran University of Medical Sciences, Tehran, IranBackground: Toxoplasmosis, a protozoan parasitic disease caused by Toxoplasma gondii, has been a serious human and veterinary medicine problem with global distribution. In the current study, we assessed immunogenicity and protective efficiency of a novel dual-promoter DNA vaccine, harboring SAG1 and GRA7 genes, from RH strain of Toxoplasma gondii (T. gondii) with or without CpG-ODN as adjuvant in a murine model.Methods: BALB/c mice were immunized intramuscularly with pVitro-SAG1-GRA7 alone and pVitro-SAG1-GRA7 with CpG-ODN three times at three-week intervals. Enzyme-linked immunosorbent assay (ELISA) was used to assess total IgG, IgG1 and IgG2a antibodies and gamma interferon (IFN-γ) and interleukin-10 (IL-10) cytokines in mice sera. Four weeks post final vaccination, MTT assay and lethal challenge-infection with 1×103 tachyzoites of T. gondii RH strain were carried out to assess stimulation index (SI) and mice survival time, respectively.Results: The IgG levels in mice immunized with multicomponent vaccines, including pVitro-SAG1–GRA7 alone and pVitro-SAG1–GRA7 with CpG-ODN, were significantly higher than those in control mice or single-gene DNA-vaccinated ones (P

Abstract Background The increased levels of blood cytokines is the main immunopathological process that were attributed to severe clinical outcomes in cases of influenza A, influenza B and people with influenza-like illness (ILI). Functional genetic polymorphisms caused by single nucleotide polymorphisms (SNPs) in inflammatory cytokines genes can influence their functions either qualitatively or quantitatively, which is associated with the possibility of severe influenza infections. The aim of the present case-control study was to investigate the association of polymorphisms in inflammatory cytokines genes with influenza patients and ILI group in an Iranian population. Methods Total number of 30 influenza B, 50 influenza A (H1N1) and 96 ILI inpatient individuals were confirmed by Real-time RT-PCR and HI assays. The genotype determination was assessed for defined SNPs in IL-1β, IL-17, IL-10 and IL-28 genes. Results The frequencies of the IL-1β rs16944 (P = 0.007) and IL-17 rs2275913 (P = 0.006) genotypes were associated with severe influenza disease, while the frequencies of IL-10 rs1800872 and IL-28 rs8099917 were not associated with the disease (P > 0.05). Also, the absence of A allele in IL-17 rs2275913 SNP increased the risk of influenza A (H1N1) infection (P = 0.008). Conclusions This study demonstrated that influenza A- (H1N1) and B-infected patients and also ILI controls have different profiles of immune parameters, and individuals carrying the specific cytokine-derived polymorphisms may show different immune responses towards severe outcome.

Multidrug-resistant (MDR) Mycobacterium tuberculosis strains, defined as resistant to at least isoniazid and rifampin, have emerged as a major worldwide health threat. Spontaneous point mutations in various genes of M. tuberculosis cause resistance to isoniazid, with the most frequent gene target being katG; and resistance to rifampin is usually due to mutation in the rpoB gene. The current study was aimed to detect the point mutations in the katG and rpoB regions related to isoniazid and rifampin resistance. A total of 203 respiratory specimens were collected from patients suspected of having tuberculosis respiratory infections referred to hospitals of Tehran, Iran, during 2018–2019. The isolation and identification of M. tuberculosis isolates were performed according to the WHO protocol. Drug susceptibility testing was carried out by proportional method. PCR analysis and sequencing were used to detect mutations in the selected katG and rpoB regions. Forty-four M. tuberculosis strains were isolated, of which 12 (27.3%) and 10 (22.7%) were resistant to isoniazid and rifampin, respectively. Ten isolates were resistant to both isoniazid and rifampin and were considered as MDR isolates. Of the ten MDR isolates, six (60%) carried mutations in both rpoB and katG. The most common mutations among isoniazid- and rifampin-resistant isolates were in codon 315 of the katG gene (70%) and codon 441 of the rpoB gene (50%), respectively. The results of this study indicated that MDR-TB continues to be a serious public health problem in Iran.

Background: One of the main obstacles to malaria control in the world has been the emergence of resistance in Plasmodium falciparum to chloroquine and other anti-malarial drugs. This study aimed to review studies in Iran on resistance in P. falciparum and P. vivax to drugs, and to reveal the mechanisms and molecular markers of resistance of these two species. Methods: The databases of PubMed, Scopus, Google Scholar, Magiran, and reputable Iranian journals were searched to find published studies on the resistance in P. falciparum and P. vivax to antimalarial drugs in Iran. Results: There is a significant relationship between resistance to chloroquine in P. falciparum and the emergence of K76T mutation in the P. falciparum chloroquine-resistance transporter gene in Iran. Resistance to sulfadoxine-pyrimethamine (SP) in P. falciparum is also significantly associated with the development of mutations in the dihydrofolate reductase and dihydropteroate synthase genes. Resistance to chloroquine in P. vivax has not been reported in Iran and it is used as a first-line treatment for P. vivax malaria. Conclusion: P. falciparum has become resistant to chloroquine in different regions of Iran and is not currently used to treat malaria. Besides, cases have emerged of P. falciparum resistance to SP in different parts of southern Iran, and SP is not administered alone for treating P. falciparum.

Abstract Background Toxoplasma gondii is a protozoan parasite with worldwide distribution, infecting a broad-range of humans and warm-blooded animals. In the current study, role of this parasite on secondary sex ratio and risk of miscarriage was investigated. Methods In this cross-sectional study, 850 cord blood samples were collected in Tehran, Iran, 2014–2015. Enzyme-linked immunosorbent assay (ELISA) was used to assess anti-Toxoplasma IgG in samples. Information such as sex of the neonates and age, number of previous pregnancies and history of miscarriage of the mothers were recorded in questionnaires. Logistic regression analysis was used to assess the possible relationship between the latent toxoplasmosis and the highlighted parameters. Results Logistic regression analysis showed that the odds of having a male neonate in seropositive women is nearly 64% higher than that in seronegative women (OR = 1.64, CI95 = 1.16–2.33, P = 0.005). The odds ratio of having male neonate increased to 2.10 (CI95 = 1.24–3.57, P = 0.006) in high-titer seropositive women, compared to that in seronegative control group. The odds of having a miscarriage history was approximately two and a half times greater in seropositive women than in seronegative ones (OR = 2.45, CI95 = 1.56–3.87, P

Rotavirus is associated with increased risk for severe diarrhea in infants and young children worldwide. This systematic review and meta-analysis was performed to determine the prevalence rate of rotavirus from different parts of Iran and provide an overall relative frequency (RF) for Iran. We performed a systematic literature review from several databases including PubMed, ISI Web of Science, Scopus, OVID, MAG IRAN, IranMedex, and Iranian Scientific Information Database. We searched the following keywords: “rotavirus,” “rotavirus infection,” “acute gastroenteritis,” “diarrhea,” “children,” “infant,” and “Iran.” The purpose of this study was to report the prevalence of rotavirus with the application of meta-analysis. We selected 43 researches out of 1147 for our study. From all the samples, the pooled estimate of prevalence (95% confidence interval) =39.9% (0.396%–0.409%) were rotavirus positive. It should be noted that rotavirus infection's RF varied from 6.4% to 79.3% in Birjand and Tehran Provinces, respectively. Thereupon, it is divergent in different studies. According to our study result, rotavirus RF has a wide range in Iran and is associated with diarrhea in children. Thus, further researches should be taken to minimize the emergence and transmission of rotavirus.

Background: Toxoplasma gondii, the coccidian protozoan parasite with worldwide distribution, is the agent of toxoplasmosis. The disease is life threatening in congenital form and in immunocompromised patients. The present study was carried out in 2016 to evaluate the in vitro effects of nanosilver colloid on tachyzoites and bradyzoites of T. gondii, RH and Tehran strains. Methods: Different concentrations (5, 10 , 20 ppm) of nanosilver colloid were added to tachyzoites of T. gondii , RH strain (type I) and bradyzoites and tissue cysts of T. gondii , Tehran strain (type II) and incubated for 30, 60, 90 and 120 minutes. The mortality rates of tachyzoites and bradyzoites were evaluated by trypan blue dye and MTT assay. Then SEM carried out to show the changes between control and exposed parasites. Results: The greatest mortality rate was seen in 20 ppm concentration and after 120 minutes of exposure. By electron microscopy, the structural changes were seen in tachyzoites of RH and tissue cyst of Tehran strain in comparison with control groups. Conclusion: Nanosilver colloid was effective on both tachyzoites and bradyzoites of T. gondii, RH and Tehran strains.

Background: Malaria is one of the most important transfusion-transmitted infections (TTIs) worldwide. To prevent the occurrence of transfusion-transmitted malaria (TTM), potential blood donors with a history of malaria or travel to, or residence in, malarious areas are permanently or temporarily deferred from donating blood. The objective of the present study was to evaluate the blood donor deferrals for malaria in Iran. Methods: This descriptive cross-sectional study was conducted in the Iranian Blood Transfusion Organization (IBTO) from 21 Mar 2011 to 19 Mar 2016 (5 yr). The data were collected and extracted from IBTO comprehensive database, namely Negareh, and then recorded in a structured template form. Statistical analysis was performed using SPSS. Results: Of the 12,790,859 blood donation volunteers, 23,084 (0.18%) were deferred due to the risk of malaria. More than 90% of malaria-deferrals were because of travel to and residence in malaria endemic areas. Among the malaria-deferred volunteers, 22,139 (95.91%) were male and 945 (4.09%) were female; 2,053 (8.89%) were permanently deferred, while 21,031 (91.11%) were temporarily deferred. The highest malaria-deferral rates were observed in South Khorasan (0.82%), Razavi Khorasan (0.79%) and Yazd (0.54%) provinces, respectively. Conclusion: Given the prevalence of malaria in neighboring countries (Pakistan and Afghanistan) and several provinces of Iran and the increasing human migration and movement between malaria non-endemic and endemic areas, the malaria-deferral rate might be higher than 0.18% in Iran. Thus, the changing, as well as the precise and accurate implementation of donor selection process must be considered in all blood transfusion centers of Iran.

An amendment to this paper has been published and can be accessed via the original article.

An amendment to this paper has been published and can be accessed via the original article.

Background: Toxoplasma gondii, cause severe medical complications in infants and immune-compromised individuals. As using early, sensitive and rapid technique has major in diagnosis of toxoplasmosis, the present study was aimed to detect parasite by using from repetitive element (RE) and B1genes, in blood samples of seropositive immunocompromised patients and pregnant women. Methods: A total of 110 peripheral blood samples were collected from seropositive cases with anti-T. gondii antibodies, including immunocompromised patients and pregnant women. DNA was extracted by a commercial kit and subjected to TaqMan probe-based real-time PCR assay by using primers and probes specific for RE and B1 genes, separately. The data were analyzed by Kappa test and SPSS-22 software. Results: In the pregnant women, 17 (68%) and 14 (56%) samples from 25 IgM+/ IgG+ cases and, 7 (25%) and 6 (21.4%) samples from 28 IgG+/IgM- cases were positive by RE and B1 real time PCR, respectively. Likewise, in immunocompromised group, 20 (66.6%) and 17 (56.6%) samples from 30 IgM+/ IgG+ cases and 2 (7.4%) and 2 (7.4%) samples from 27 IgG+/ IgM- cases were positive by RE and B1 real time PCR, respectively. Conclusion: Probe-based real time PCR assay is a quantitative approach for early diagnosis of T. gondii infection in clinical samples. Moreover, this method can be more appropriate in diagnosis of acute and reactivated toxoplasmosis. In addition our results indicated that RE gene is more sensitive than B1 gene.

Background: Vivax malaria is more prevalent in the malarious areas of Iran, which makes vaccine research a high priority. Serine Repeat Antigens (SERA) have essential role in the parasite life cycle and high expression profiles of PvSERA5 make it suitable vaccine candidates. This study aimed to evaluate the genetic diversity of C-terminal region of PvSERA5 in Iranian isolates of Plasmodium vivax in Sistan and Baluchistan. Methods: Totally, 49 blood samples were taken from symptomatic malaria patients in Sistan and Baluchistan Province in 2016. Mono-infection to P. vivax was confirmed by 18srRNA-Nested-PCR. Genomic DNA was extracted and C-terminal region of PvSERA5 was amplified by specific primers. PCR-products have been sequenced and analysis was done by using bioinformatics software, mainly DnaSP & MEGA5. Results: Genetic diversity was calculated 14.8% in C-terminal region of PvSERA5 in Iranian isolates, 19 different sequences and 4 haplotypes existed. The amount of Tajima’s D (0.3805) and ratio of non-synonymous to synonymous mutation (1.82) showed that C-terminal region of PvSERA5 is under positive natural selection; also intragenic recombination could interfere. Conclusion: Results could be helpful in any research, regarding this antigen as vaccine candidate in Iran or worldwide.

Background: Cutaneous leishmaniasis (CL) is one of the world health problems. Leishmania major is the etiological agent of zoonotic cutaneous leishmaniasis (ZCL). Promastigote and amastigote are two morphological forms of Leishmania parasites that express different proteins and p27 is an important gene encoding cytochrome c oxidase (COX) component. P27 gene expresses a 27 kDa protein that essential in ATP synthesis. This study aimed to compare p27 gene expression in promastigote and amastigote forms in Iranian strain of L. major (MRHO/IR/75/ER). Methods: This study was conducted in 2015. Clinical isolates of CL patients from north, center, west and south parts of Iran were collected and identified by PCR-RFLP. After RNA extraction of promastigotes and amastigotes and cDNA synthesis, the expression level of p27 gene was compared by real-time RT-PCR. Results: By comparison of expression level between amastigote and promastigote forms of Iranian strain of L. major, up-regulation of p27 gene (2.73 fold) was observed in amastigotes. Moreover, there was no significant difference in p27 gene expression between L. major isolates. Conclusion: p27 gene and protein can be considered as a target in recombinant vaccine production and treatment process.

Background: Although rigorous efforts have substantially decreased the malaria burden through decades, it still threatens the lives of millions of children. Development of an effective vaccine can provide important approach in malaria control strategies. Unfortunately, development of an effective vaccine for falciparum malaria has been hindered by the extreme complexity of malaria parasite biology, complex and diverse parasite genomes, and immune evasion by the parasites as well as the intricate nature of the parasites infection cycle. The aim of this review was to discuss the different approaches to malaria vaccine development until now. Methods: Scientific databases, including MEDLINE (via PubMed) and SCOPUS were searched up to 30 Jan 2017 and the articles regarding malaria vaccine development were taken into examination. Results: Several strategies for malaria vaccine development including pre-erythrocytic vaccines, antibody-based subunit vaccines, vectored vaccines, whole sporozoite vaccines, genetically Attenuated parasites and sporozoite subunit vaccine, erythrocytic vaccines, sexual stage vaccine, transmission-blocking vaccine as well as synthetic peptides and conjugate vaccine has been introduced. However, the success has been limited thus far. Conclusion: Although development of malaria vaccine over the past 70 year has been continued, the discovery, development, and licensing of a malaria vaccine formulation, which meets safety, affordability, accessibility, applicability, and efficacy has not yet been achieved.

Background: Visceral leishmaniasis (VL) has strong links with poverty, substantial medical and veterinary impacts. This review aimed to focus in studies published during 1994-2016 on VL in southeastern Iran. Methods: The present review is based on expert knowledge and historical studies published during the past 23 yr (1994-2016) on VL in southeastern Iran. In addition, related literature found in PubMed by using the keywords such as visceral leishmaniasis, kala-azar, and Leishmania infantum are included. Results: Overall, 118 children aged 4.2 yr were detected as infected with human VL (HVL). The majority of the cases were from Orzoieh district (37.1%) in southwest of Kerman Province, followed by Sirjan (15.7%), Jiroft (14.8%), Kahnuj (9.3%) and to lesser extent from other areas. The male to female ratio was 1.7. The three most frequent clinical features were represented by fever (100.0%), anemia (95.0%) and splenomegaly (91.5%). Altogether, 42.0% of the VL cases developed secondary bacterial infections, the overall case-fatality rate was 3.4%, and majorities (88.0%) of the VL patients were undernourished. Overall, 733 dogs and wild canines were examined by different techniques with various seroprevalence ranges. Conclusion: In southeastern Iran, VL is endemic in Orzoieh district in Kerman Province. While the dogs are implicated as the main domestic reservoir of VL, wide range of wild canines can serve as a secondary potential reservoir host.

Background: Here, we established the mouse models of chronic toxoplasmosis by T. gondii Tehran strain to provide a good understanding about defining the possible association between T. gondii exposure and learning and memory impairments. Moreover, as secondary objective of the present study, we hypothesized whether administration of an acetylcholinesterase (AChE) inhibitor could reduce learning and memory impairments induced by T. gondii infection. Methods: Twenty-four male BALB/c mice were used to establishment of latent toxoplasmosis. The animal model of Toxoplasma infection was established by the intraperitoneal inoculation of 20-25 tissue cysts from Tehran strain of T. gondii. Donepezil (2 mg/kg) an AChE inhibitor to treat Alzheimer disease was injected intraperitoneally once a day for two weeks starting from post-infection day 90. Morris water maze (MWM) task was used to assay spatial learning and short term spatial memory in all groups. One-way ANOVA with Tukey’s post-hoc test was used to assess differences between experimental groups. P

Background: Malaria is the most important transfusion-transmitted infection (TTI) in worldwide after viral hepatitis and human immunodeficiency virus (HIV) infection. The main objective of the present study was to review and evaluate the transmission of malaria via blood transfusion in Iran. Methods: A literature search was done without time limitation in the electronic databases as follows: PubMed, Scopus, Google Scholar, Web of Science, Science Direct, scientific information database (SID), Magiran, IranMedex and Irandoc. The searches were limited to the published papers to English and Persian languages. Results: Six papers were eligible. From 1963 to 1983, 344 cases of Transfusion-transmitted malaria (TTM) had been reported from different provinces of Iran. The most prevalent species of involved Plasmodium in investigated cases of TTM was Plasmodium malariae (79.24%). The screening results of 1,135 blood donors for malaria were negative by microscopic examination of peripheral blood smears and rapid diagnostic test (RDT) methods. Conclusion: Lack of TTM report from Iran in the last three decades indicates that the screening of blood donors through interviewing (donor selection) may be effective in the prevention of the occurrence of transfusion-transmitted malaria.

Background: As a waterborne pathogen, Cryptosporidium is one of the most common causes of gastroenteritis in human and hoofed livestock animals. This study aimed to investigate the distribution of Cryptosporidium spp. in human and livestock wastewaters in Iran, by the 18S rRNA sequence analysis. Methods: A total of 54 raw wastewater samples collected from three urban treatment plants and two slaughterhouses during 2014-2015 in Tehran, Iran. The presence of the Cryptosporidium oocysts was assessed by immunofluorescence with monoclonal antibodies. To characterize the oocysts at the molecular level, the 18S rRNA gene of Cryptosporidium was PCR amplified and sequenced. Results: Of the 54 wastewater samples examined, 34 (62.9%) were positive for Cryptosporidium oocysts using the IFA. Of these, 70.5% (24/34) were positive by PCR, that 91.6% (22/24) were successfully sequenced. The species of C. andersoni (95.4%) and C. xiaoi (4.6%) were detected in livestock wastewater samples. Conclusion: C. andersoni was the major Cryptosporidium sp. found in the aquatic environmental wastewater samples. The high rate of detection of C. andersoni in domestic wastewater was probably the result of the predominancy of this species in cattle herds in Iran. The current study is the first report of C. xiaoi in Iran.

Background: We aimed to quicken and simplify the detection of Plasmodium in blood samples by developing and testing a pan-Plasmodium real-time PCR for accurate screening of individuals suspected of malaria. Methods: A single primer/probe set for pan-species Plasmodium-specific real time PCR targeting a conserved region of the small subunit 18S ribosomal DNA was designed and evaluated for rapid diagnosis and screening of malaria infections using dried blood smears. FTA cards were used for rapid and simple DNA extraction. Results: The primers and probes showed a positive response with the DNA extracted from bloods infected with P. falciparum and P. vivax but not with DNA extracted from various smears from uninfected blood samples. Seven positive cases positive by both microscopy and nested PCR were found among 280 blood samples taken from in South and Southeast Iran. Five samples were identified as positive for P. vivax and two as positive for P. falciparum. All positive samples were positive by real-time PCR. Furthermore, all 38-blood samples positive by microscopy were positive by real-time PCR. No microscopy-negative samples were positive by real-time PCR. Conclusion: By using a simple FTA card for DNA extraction and by application of the real-time PCR developed in this study, sensitivity similar to nested-PCR and microscopy was achieved. This format simplifies the detection of Plasmodium in large numbers of samples.

Background: Based on recent studies, there are controversial reports on the capac­ity of tissue cyst forming of Toxoplasma gondii RH strain. In this study, the capac­ity was evaluated by in vivo and in vitro experiments.Methods: RH strain was subcutaneously inoculated to ten Wistar rats. After one month, their blood, brain, tongue and diaphragm were collected and evaluated by MAT, PCR, pathological and bioassay methods. The parasite was cultivated in the cell monolayer. To change to bradyzoite, the media pH was altered to 6.8. Biologi­cal aspect of the bradyzoites was evaluated by incubation in acidic pepsin and it's inoculation in ten BALB/c mice.Results: All rats showed antibodies to Toxoplasma at titers ≥1:320 but no DNA and tissue cyst were detected in the tissues. Following intraperitoneal inoculation of rats’ brain homogenate into BALB/c mice, no infection was established in none of the animals. During presence of cell culture, in acid media for a 3-5 days period, cyst-like structures were noticed when they were stained with PAS. The visible brady­zoites in the cysts that were incubated in acid pepsin medium were not able to kill any mice.Conclusion: This study confirmed that Iranian RH strain has lost the potential of tissue cyst forming in rats and bradyzoites cultivated in cell culture lost their resis­tance to acidic condition, so this strain can be a candidate for future vaccine re­searches.

Background: Toxoplasma gondii is an obligate intracellular parasite that infects humans at high preva­lence rates. The virulent RH strain of T. gondii is generally considered to have lost its cyst forming capacity. This study performed to obtain tissue cysts in mice infected with tachyzoites of RH strain treated with sulfadiazine (SDZ). It provides the opportunity to analyze the conversion of tachyzoite to bradyzoite stage of the RH strain, followed by stage-specific gene-expression analyzing.Methods: Two groups of Swiss-Webster and BALB/c mice were infected subcutaneously with 104 tachyzoites of T. gondii, RH strain and given SDZ (300 mg/l) with NaHCO3 (5 g-1) in drinking water from day1 to day 14 post infection (p.i). The infected mice were sacrificed on day 50 post infection. Their brains were removed and the numbers of tissue cysts were microscopically counted. Total RNA was extracted from brains and cDNA synthesis was carried out. Finally, RT-PCR (Reverse transcrip­tion PCR) was used to detect the expression of bradyzoite (BAG1) and tachyzoite (SAG1) specific genes during tachyzoite / bradyzoite stage conversion.Results: Sixty five percent of all infected mice were survived. Cysts were detectable in mice brain (45%) on day 50 p.i. Also RT-PCR of the brain samples was positive for SAG1 and BAG1.Conclusion: It seems that conversion of tachyzoites to bradyzoites in brain of mice undergoing SDZ was not completed until 50 days after inoculation.

Background: The main goal of present study was to detect polymorphism in MSP-1 gene which is a major blood stage candidate for vaccine in Plasmodium vivax by Single Strand Conformational Polymor­phism-Polymerase Chain Reaction (SSCP-PCR).Methods: During 2008 to 2010 fifty samples were collected from Iranian patients with P. vivax in Hormozgan Province, southern Iran. All of the samples were detected by microscopical examination. Amplification of MSP-1 gene was done by PCR after DNA extraction. Single strand DNAs due to using in SSCP, was electrophoresed on polyacrylamid- Bisacrylamid gel then banding patterns were revealed by silver-staining method. Sequencing as a typing method was performed for some isolates.Results: All of the 50 isolates were positive microscopically. Totally 12 (24%) isolates showed 440 bp and 38 (76%) showed 500 bp in PCR assay. SSCP analysis revealed four banding patterns. Pattern I (10/50), Pattern II (12/50), Pattern III (27/50), and Pattern IV (1/50). The results sequencing analy­sis of the MSP-1 gene in 19 isolates revealed diversity in nucleotides and amino acid in Iranian P. vivax isolates.Conclusion: Our study confirms that the SSCP-PCR is a rapid method for detecting polymorphism in MSP-1 gene in P. vivax. The presence of different haplotypes in MSP-1 gene shows that several P. vivax strains exist in malaria endemic areas of Iran.

Background: This study was performed to induce conversion of RH strain tachyzoites of Toxoplasma gondii to bradyzoites by pH changing of the cul­ture medium. Methods: HeLa cell monolayers were infected at a 1:1 tachyzoite to cell ratio. Four hours after infection, the culture medium was removed and re­placed with culture medium and 5% FCS, adjusted to pH 8 with NaOH. The culture was maintained at 37 °C without CO2 until the end of the ex­periment. Cyst-like structures were collected and stained with periodic acid schiff (PAS) staining method. The soluble antigens of cyst-like structures of RH strain, in addition to RH tachyzoite, bradyzoites of avirulent Tehran strain and uninfected HeLa cells were electrophoresed on 12.5% poly­acrylamide gel. The gel was stained by coomassie brilliant blue R-250. Results: Four days after infection of HeLa cells with tachyzoites of T. gondii, RH strain, cyst- like structures were noticed and stained with PAS. In the SDS-PAGE, protein bands of these structures had some differences with tachyzoites of RH strain, but there was quite similarity between pro­tein bands of these structures and tissue cysts (bradyzoites) of Tehran strains. P34 and P36 (bradyzoite-specific proteins) were observed only in T. gondii bradyzoites of RH (cyst like structures) and bradyzoites of Tehran strains. Conclusion: Alkalization of culture medium to pH 8 induced expression of bradyzoite- specific proteins and production of RH cysts in cell culture.

Background: The assays currently available for the detection of specific anti-Toxoplasma antibodies may vary in their abilities to detect serum immunoglobulins, due to the Lack of a purified standardized antigen. The aim of this study was evaluation the recombinant Toxoplasma gondii SAG1 antigen for the serodiagnosis of acute and chronic toxoplasmosis. Methods: This study describes an ELISA using recombinant SAG1 for detection of IgM and IgG antibodies against Toxoplasma gondii in human sera. Genomic DNA of T. gondii (RH Strain) was isolated and PCR reaction was performed. Recovered DNA was cloned into PTZ57R cloning vector. The recombinant plasmid was detected by restriction analysis. The SAG1 gene was subcloned in the pET- 28a expression vector. Protein production was then induced with 1 mM isopropyl-D - thiogalactopyrano­side (IPTG). A total of 204 sera were tested using a commercial IgG and IgM ELISA kit (Trinity, USA) as gold standard prior to testing them with the recombinant antigen. Results: Tested sera were divided into the following groups:(a) The 74 T. gondii IgG positive (b) 70 T.gondii IgM positive (c) 60 sera who had no serological evidence of toxoplasmosis as negative sera.To determine the specificity of the test, we used other parasitic diseases including echinococusis (N=5), malaria (N=14), leishmania­sis (N=7),fasciolasis (N=4 ), sterengyloidiasis (N=1 ). Sensitivity and specificity of the generated recombinant IgG ELISA in comparison with commercial ELISA (Com ELISA) were 93% and 95%, and the sensitivity and specificity of the generated recombinant IgM ELISA were 87% and 95% respectively. Conclusion: The results acquired here show that this antigen is useful for diagnostic purposes and could be replaced by lysed, whole cell antigens for diagnosis of chronic toxoplasmosis.

Background: Hydatidosis is one of the major zoonotic diseases that cause considerable public health problems in Iran. The present study was designed to investigate pediatric hydatidosis in patients referred to the Children Medical Center Hospital in Tehran, Iran during 2005-2010.Methods: Data were collected from the records of 17 patients referred to the center with hydatidosis. Data included demographic data; laboratory results, type, and site of cysts, clinical manifestations, and treatment.Results: Nine patients were boys (52.9 %) and eight (47.1 %) were girls. Most patients referred from central areas of Iran (58.8%). Seven patients had cysts in their lungs (41.2%) and three cases (17.6 %) in liver. Six cases (35.3 %) had simultaneous lung and liver cysts, 3 patients (17.6%) had brain cysts (alone or in combination with other organs involvement) and 2 patients (11.7%) showed multi-organ involvement. All patients were treated by albendazole and underwent surgery, recurrence was seen in 4 (23.5 %) of the cases and one patient died due to rupture of the cyst and anaphylactic shock.Conclusion: Multi-organ involvement seems to be on the rise in children, this has led to the necessity for physicians to be more aware of clinical features, search, and rule out other organs for involvement diagnosis once a cyst is detected in one organ.

Background: Plasmodium vivax is the predominant species causes of malaria with about 90% total annual reported malaria in Iran. This study conducted to determine the susceptibility of Plasmodium vivax isolates to chloroquine in Sistan and Balochistan Province, southeastern Iran.Methods: A total 270 subjects with symptomatic malaria and confirmed P. vivax infection completed the designed 28-day in vivo study. The thick and thin film blood smears were screened for malaria parasites by microscopy. The nested PCR was applied using the Plasmodium 18 subunit ribosomal ribonu­cleic (Ssr RNA) genes for detecting mixed infections and diagnosis of parasites in the samples with low parasite on days 0, 5, 6, 7, and 28. Results: P. vivax was cleared in 15%, 50%, 95%, and 100% of patients on days 1, 2, 3, 4 respectively by microscopy assessment. Six patients were exhibited specific P. vivax band in nested PCR on day 5. No recurrence was observed on days 7, 14 and 28. Mean (±standard deviation) parasite clearance time was 2.41 (±0.8) days. Conclusion: P. vivax is still susceptible to chloroquine in Southeatern Iran. This finding is compati­ble with results of neighboring countries Pakistan and Afghanistan.

Background: Cryptosporidium is a worldwide protozoan parasite and one of the most common causes of infection and diarrhea in humans and cattle. The aim of the present study was determina­tion of subtypes of Cryptosporidium among children with diarrhea in Tehran by se­quence analysis of the highly polymorphic 60-kDa glycoprotein (GP60) gene.Methods: Fecal samples were collected from 794 diarrheic children. Initial identification of Crypto­spo­ridium was carried out on stool samples by Ziehl-Neelsen acid-fast staining method. DNA was extracted from positive microscopically samples and Cryptosporidium genotypes and subtypes were determined, accordingly."nResults: Out of 794 collected samples, 19 (2.40 %) were positive for Cryptosporidium oocysts. Sequences analysis of GP60 gene showed that 17 (89.47 %) of the positive isolates were Crypto­spori­dium parvum and 2 (10.52 %) were C. hominis. All subtypes of C. parvum isolates belonged to allele families IIa (6/17) and IId (11/17). The most common allele in all 17 isolates belonged to IId A20G1a (41.18%). A22G1 (IF) subtype was detected in two C. hominis isolates of the chil­dren."nConclusion: The predominancy of C. parvum species (specially, IId A20G1a sub­type) in current study underlines the importance of zoonotic Cryptosporidium transmission in Iran.

Background: Schizophrenia is a serious, chronic, and often debilitating neuropsychiatric disor­der. Its causes are still poorly understood. Besides genetic and non-genetic (environmental) fac­tors are thought to be important as the cause of the structural and functional deficits that character­ize schizophrenia. This study aimed to compare Toxoplasma gondii infection between schizo­phrenia patients and non-schizophrenia individuals as control group.Methods: A case-control study was designed in Tehran, Iran during 2009-2010. Sixty-two pa­tients with schizophrenia and 62 non-schizophrenia volunteers were selected. To ascertain a possible relationship between T. gondii infection and schizophrenia, anti-Toxoplasma IgG antibodies were detected by indirect-ELISA. Data were statistically analyzed by chi- square at a confidence level of 99%.Results: The sero-positivity rate among patients with schizophrenia (67.7%) was significantly higher than control group (37.1) (P

Background: Toxoplasmosis is a worldwide endemic disease. In congenitally infected infants and AIDS patients, toxoplasmosis causes high rates of morbidity and mortality. In these cases antibody detection is difficult; so detection of parasite or its components could be useful tool for early detection and following treatment of the infection.Methods: Sixty-three BALB/c mice were injected intra-peritoneal with 5×103 tachyzoites of Toxoplasma gondii RH strain, nine mice were sacrificed daily for 7 days. Fourteen mice were in­jected with phosphate buffer saline as control group. Dot-ELISA was performed for detection of T.gondii antigen in mice sera and capture - ELISA was done as golden standard assay too.Results : Toxoplasma gondii antigen was detected from day 2 in mice sera ; 22% of mice sera on day 2, 33% on day 3,77% on day 4 and 100% on day 5 till their death on day 7 had shown antigene­mia by dot - ELISA, no positive result was detected in control mice by dot- ELISA.Conclusion: Dot-ELISA is a sensitive method for diagnosis of T. gondii infection in the animal model; also, this technique is more rapid and easy to perform method in comparison with cap­ture-ELISA.