Molecular Identification and Phylogenetic Analysis of Leishmania major Isolated from Zoonotic Cutaneous Leishmaniasis Foci in Northeast Iran.

Background: Epidemiological studies, classification and genetic studies of Leishmania species are effective in treatment, control and prevention in endemic areas. We aimed to investigate the genetic diversity and phylogeny of Leishmania in Zoonotic foci located in northeastern Iran using nagt gene for the first time. Methods: DNA of 100 confirmed positive slides collected from the health centers of Sarkhes, Darghez, Fariman, Esfarayen, and Sabzevar were extracted during 2020-2021. The partial sequence of kDNA was amplified to identify the species. Twenty-five DNA samples were randomly subjected to amplify by nagt gene primes and were sequenced. The sequences were aligned with reference sequences in National Center for Biotechnology Information (NCBI). Then, the genetic similarities of the sequences were checked using Clustalx2.1 software and the phylogenetic tree was drawn by Mega 7 software. Results: All the positive samples were diagnosed as L. major. Approximately, half of the sequences of species were similar to two reference genes JX103550.1:404-712 L. major Esfahan and KX759012.1:568-807 L. Major Ilam (more than 90% similarity). According to the results of the phylogeny tree, the closest genotype to our study samples was JX103550.1:404-712 L. major Esfahan. Conclusion: The most causative agent CL in these areas was L. major. The genetic diversity of L. major was high such as other zoonotic foci in Iran. Due to the high similarity of the strains in the study areas with the strains of Isfahan and Ilam, similar control and prevention methods is suggested in these areas.

Development of a Novel HTLV-1 Protease: Human Fcγ1 Recombinant Fusion Molecule in the CHO Eukaryotic Expression System.

Human T-cell leukaemia virus type 1 (HTLV-1) is the causative agent of two life-threatening diseases, adult T cell leukaemia/lymphoma (ATLL), and HTLV-1-associated myelopathy/tropical spastic (HAM/TSP). HTLV-1 protease (HTLV-1-PR) is an aspartic protease that represents a promising target for therapeutic purposes like human immunodeficiency virus-PR inhibitors (HIV-PR). Therefore, in this study, the human Fc fusion recombinant-PR (HTLV-1-PR:hFcγ1) was designed and expressed for two applications, finding a blocking substrate as a potential therapeutic or a potential subunit peptide vaccine. The PCR amplified DNA sequences encoding the HTLV-1-PR from the MT2-cell line using specific primers with restriction enzyme sites of Not1 and Xba1. The construct was then cloned to pTZ57R/T TA plasmid and, after confirming the PR sequence, subcloned into the pDR2ΔEF1α Fc-expression vector to create pDR2ΔEF1α.HTLV-1-PR:hFcγ1. The integrity of recombinant DNA was confirmed by sequencing to ensure that the engineered construct was in the frame. The recombinant fusion protein was then produced in the Chinese hamster ovary cell (CHO) system and was purified from its supernatant using HiTrap-rPA column affinity chromatography. Then, the immunofluorescence assay (IFA) co-localisation method showed that HTLV-1-PR:hFc recombinant fusion protein has appropriate folding as it binds to the anti-Fcγ antibody; the Fcγ1 tag participates to have HTLV-1-PR:hFcγ1 as a dimeric secretory protein. The development and production of HTLV-1-PR can be used to find a blocking substrate as a potential therapeutic molecule and apply it in an animal model to assess its immunogenicity and potential protection against HTLV-1 infection.

Examining the Relationship between Salivary Amylase Level, Head Trauma Severity and CT Scan Results in Patients with Isolated Mild Head Trauma.

Objective: To investigate the relationship between salivary amylase level and computed tomoraphy (CT scan) findings in patients with isolated mild traumatic Brain Injury (mTBI) referred to the emergency department of Shahid Hasheminejad Hospital. Methods: Patients with isolated mTBI and indication for brain CT scan who referred to the trauma center of Shahid Hasheminejad Hospital, Mashhad, Iran in 2019 were included in a cross-sectional study. In the initial examination, the patient's level of consciousness was measured using the Glasgow Coma Scale (GCS), and saliva samples were taken at the emergency department to determine the level of salivary amylase. A brain CT scan was performed for all patients. Age, gender, cause of trauma, the trauma severity and CT scan results were recorded. Statistical analysis was performed on the data. Results: One-hundred fifty patients were enrolled in this study (men=101, women=49). The trauma causes were included accidents (n=88; 58%), falls (n=37; 25%) and miscellaneous factors (e.g., quarrels; n=25; 17%). GCS was 15 in 142 patients and 14 in the rest. In all patients, the trauma severity was mild to high risk (Minor). CT scan results unfolded pathology in 10 cases (7%), while the residues (93%) had normal CT scans with no pathological evidence. Salivary amylase level in the patients' saliva samples was between 137 to 8000 units per liter. Using the t-test to evaluate the relationship between salivary amylase levels and CT scan results uncovered a significant relationship. Spearman correlation revealed no significant relationship between the amylase and GCS levels. Conclusion: Data statistical analysis from 150 patients with isolated head trauma manifested that salivary amylase levels were significantly higher in the patients with pathological findings on CT scans. However, no significant relationship was found between salivary amylase level and age, gender, cause of trauma, and level of consciousness.

Local and systemic gene expression levels of IL-10, IL-17 and TGF-ß in active ocular toxoplasmosis in humans.

BACKGROUND: To compare mRNA expression of interleukin 10 (IL-10), interleukin 17 (IL-17) and Transforming Growth Factor-ß (TGF-ß) in aqueous humor (AH) and peripheral blood mononuclear cells (PBMCs) in human ocular toxoplasmosis (OT) and controls. METHOD: RNA isolation, cDNA synthesis and real-time polymerase chain reaction were performed on AH sediments and PBMCs of 16 patients with active OT and 21 controls at the Khatam-al-Anbia Eye Hospital, Iran. For comparison, Mann Whitney U test was used at a discrimination level of p < 0.05. Pearson and Spearman rank correlation test were applied for correlation with clinical parameters. RESULTS: The expression for IL-10 and IL-17 in the AH was 3.7- and 88.0-fold higher in OT than in controls (P = 0.04 and P = 0.03, respectively) whereas that of TGF-ß was 7.7-fold lower (P < 0.001). The expression levels for these cytokines in PBMC followed a similar pattern (IL-10 13.8-fold down-regulated (P = 0.001), IL-17 with 1.9-fold insignificantly upregulated (p = 0.43), TGF-ß 452.8-fold down-regulated (P = 0.002). Compared to PBMC, IL-10 coding mRNA was 1876-fold higher in the almost cell-free AH in OT (39.2-fold in controls), IL-17 coding mRNA was 9.4-fold higher (17.7-fold down-regulated in controls), and that coding for TGF-ß 207-fold higher in OT (7x105-fold in controls). The expression for IL-10, IL-17 and TGF-ß in AH thus followed an opposite pattern compared to that in PBMC. CONCLUSION: OT induces a highly-specific local immunoregulatory process as evidenced by an intraocular up-regulation of IL-10 and down-regulation of TGF-ß mRNA. This could indicate an attempt to prevent unnecessary tissue damage which is in line with a moderate local mRNA up-regulation for IL-17 which seems sufficient to control parasite proliferation. That this regulation is opposite to that in PBMC may be linked to intraocular immune deviation in the course of disease.

Are certain factors involved in calcium metabolism associated with temporomandibular disorders?

Objective: The aim of the present study was to investigate whether there is an association between serum concentrations of calcium, phosphate, alkaline phosphatase, parathyroid hormone (PTH), and vitamin D and temporomandibular disorders (TMDs).Methods: In this case-control study, 51 patients with TMDs (28 patients with TMJ clicking, 5 patients with muscular disorders, and 18 osteoarthritis patients) and 29 healthy subjects within the age range of 20-50 years as the control group, were referred to the laboratory for calcium, phosphate, alkaline phosphatase, PTH, and vitamin D (25 OHD) analysis tests. The TMJ status of all participants was examined according to RDC-TMD criteria by a prosthodontist who is an expert in TMDs. The results were analyzed with Fisher's precise test.Results: No statistically significant difference was observed between TMD patients and healthy subjects.Conclusion: Serum concentrations of calcium, phosphate, alkaline phosphatase, PTH, and vitamin D are not associated with TMDs.

Communication barriers between basic scientists and clinicians in regenerative medicine: A qualitative study from Iran.

RATIONALE, AIMS, AND OBJECTIVES: Cell-based therapeutics are among the latest advances in health care technologies. The rapid evolution of stem cell science in Iran has necessitated the application of scientific achievements in clinical settings. However, various issues hindered their translation, in particular, impediments in the interactions of basic stem cell scientists and clinicians. We highlighted the impediments in the interactions of stem cell scientists and physicians involved in the opinion of professionals from both groups. METHOD: This qualitative research was conducted with thematic analysis, performed by purposive sampling. Thirty-two distinguished stem cell scientists and clinicians were interviewed to identify their perspectives on this matter. MAXQDA 2018 was used to classify the axial codes based on factors related to communications inefficiencies. The analysis of coded data recognized 18 subthemes and six major themes. RESULTS: Central themes include different registers of the two parties, counterproductive clusters hampered networking, external communication barriers, the competition to access resources, leadership conflicts, and the dissatisfaction of stakeholders with their share. CONCLUSIONS: Most of the impediments were seemingly global, for example, the incoherent medical and basic science educational systems, the vulnerable career path of physician-scientists, and an increasing tendency towards overspecialization. However, some local specific issues were also described, for example, limited funding opportunities and the negative impacts of the division of medical education from the ministry of science, research, and technology in Iran. Proposed interventions include the reinforcement of physician-scientist programs, designing a distributed leadership model, and bringing back the scientific integrity to higher education in Iran.

Clinical and Laboratory Findings of Visceral Leishmaniasis in Children Hospitalized in Mashhad, Northeastern Iran: A Twenty-Year Retrospective Study.

BACKGROUND: Over the last decade, a few cases of visceral leishmaniasis (VL) have been reported in some provinces of northeastern Iran. We aimed to investigate clinical and laboratory findings of VL among children who admitted to the pediatric ward in a referral hospital in Mashhad, northeastern Iran. METHODS: A retrospective study, between 1997 and 2017, was performed on the data sheet registered for children with confirmed VL at the referral Emam Reza Hospital in Mashhad. Hematological and biochemical profiles of the patients were analyzed. RESULTS: A total of 35 children with VL, confirmed by the presence of amastigotes of Leishmania in Giemsa stained smears of the bone marrow, had been recorded through 20 yr. The mean age of patients was 3.7±4 yr. The majority of the patients suffered from hepatosplenomegaly (100%, n=35/35), followed by prolonged fever and pallor (91%, n=32/35), weight loss (85%, n=30/35). The main laboratory findings were anemia (94.1%), leukopenia (52.9%) and thrombocytopenia (70.5%). Almost one-third (37.1%; 13/35) of VL patients inhabited in rural areas of the Bojnoord district as a known VL endemic focus in northeastern Iran. CONCLUSION: Our preliminary data showed that the origin of VL is still in some districts other than Mashhad, where VL just will be diagnosed.

IgG Avidity Test for Ocular Toxoplasmosis Diagnosis at a Tertiary Center, Northeast of Iran.

BACKGROUND: The diagnostic methods which are used for acute ocular toxoplasmosis are very important; if the treatment is delayed, it sometimes leads to loss of vision. Few studies have been performed to evaluate serological tests used in the diagnosis of acute ocular toxoplasmosis. OBJECTIVES: To evaluate the immunoglobulin (Ig) M, G and IgG avidity tests for diagnosis of acute ocular toxoplasmosis in the northeast of Iran. METHODS: A cross-sectional study was carried out from January 2014 to December 2016. After an opthalmic examination was conducted by a retina specialist, 16 typical acute and 34 typical chronic ocular toxoplasmosis cases were included in this study. Information on clinical manifestations, age and occupation was recorded. Anti-Toxoplasma IgG, IgM and IgG avidity tests were administered on serum samples using the ELISA method. RESULTS: Blurring of vision in all patients was the most clinical presentation. The IgG avidity test could diagnose all acute and recent cases. However, three false positive and one false negative result occurred using the IgM test by ELISA. The false negative result in all likelihood occurred because the patient was at the beginning stage of the infection. CONCLUSION: The result of this study showed that IgM is not a reliable marker of acute disease. Repetition of the serology tests was proposed in cases with clinical manifestations without detectable antibody titer after approximately two weeks. IgG avidity testing results coincided with clinical diagnosis and it could therefore considered to be a reliable method to differentiate between recently acquired and chronic ocular toxoplasmosis.

Diagnosis of acute toxoplasmosis by IgG avidity method in pregnant women referred to health centers in south-eastern Iran.

Toxoplasmosis, one of the most common parasitic infections, can cause abortions in human. The purpose of this study was to determine seroprevalence of toxoplasmosis and acute form of toxoplasmosis in pregnant women. 208 pregnant women who referred to health centers in south-eastern Iran were taken under study after signing the informed consent forms and filling out the checklists. For those with high levels of IgG or IgM antibody titers, IgG Avidity test was performed to determine the acute infection. High level of IgG anti-body were found in sera of 81 pregnant women and 7 persons had borderline titer. IgM antibody results were positive in 33 and borderline in 4 cases. Then, the samples with positive and borderline results for IgM and IgG were evaluated by IgG avidity test. In this study, approximately 60% of pregnant women were not immune to risks posed by Toxoplasma gondii. Therefore, this group are at risk of acquiring primary infection of toxoplasmosis during gestation.

Genotyping of Echinococcus granulosus Isolates from Human in Khorasan Province, North-Eastern Iran.

BACKGROUND: Human hydatidosis is endemic in northeastern Iran. The present study aimed to investigate molecular diversity of Echinococcus granulosus isolates collected from human surgically. METHODS: Sixty human hydatid cysts (58 lung cysts and 2 liver cysts) were collected through surgery from Ghaem and Emam Reza hospitals in Mashhad University of Medical Sciences during 2015-2016. Cysts were characterized using polymerase chain reaction-restriction fragment length polymorphism (PCRRFLP) analysis of the internal transcribed spacer 1 (ITS1) gene and sequencing fragments of the genes coding for mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit I (nad1). RESULTS: Overall, 55 out of 60 Echinococcus granulosus cysts (91.6%) were determined as the G1 strain, 4 cases (6.6%) were determined as the G6 strain and 1 sample was not identified. CONCLUSION: Although sheep strain (G1) is dominated in human patients in Great Khorasan, the prevention of camel-dog cycle should pay attention in this region.

Investigation of Visceral Leishmaniasis among 192 Dog Carcasses Killed by Road Accidents in Khorasan Razavi, Northeastern Iran during 2014-2016.

BACKGROUND: Visceral leishmaniasis (VL), so-called Kala-azar is a life threating parasitic infectious disease caused by Leishmania spp. L. infantum is the main causative agent for Mediterranean form of Kala-azar which is endemic in northeastern Iran. This study attempted to investigate existence of canine visceral leishmaniasis (CVL) in Khorasan Razavi. METHODS: Between 2014 and 2016, tissue samples collected from spleen and liver of 192 stray dogs were examined to investigate existence of L. infantum. Kinetoplast DNA (k-DNA) PCR was performed to identify the species of parasites. The positive PCR products were sequenced in both directions to confirm the kDNA PCR results. RESULTS: Among samples obtained from 192 dogs, kinetoplast DNA of L. infantum was detected in two female dogs. L. infantum was confirmed by sequence analysis of PCR products. CONCLUSION: Our data confirm stray dogs play as potential reservoirs for VL in this province. Further investigation will be necessary to clear role of stray dogs in the transmission of L. infantum to human and domestic dogs.

Correction to: Assessment of ocular toxoplasmosis patients reported at a tertiary center in the northeast of Iran.

Unfortunately, in the original publication of the article, given name and family name of the first and fourth author names were mentioned incorrectly. The exact given name and family name of those author names have been provided in the erratum.

Assessment of ocular toxoplasmosis patients reported at a tertiary center in the northeast of Iran.

PURPOSE: Ocular toxoplasmosis, which is caused by the single-cell parasite Toxoplasma gondii, is currently the most significant cause of posterior uveitis in the world. No previous studies have described the prevalence and clinical features of ocular toxoplasmosis in the northeast of Iran. The purpose of the current study was to address this gap. METHODS: In this retrospective study, the medical records of 488 uveitis patients who presented to the Khatam-al-Anbia Eye Hospital of Mashhad University of Medical Sciences, a tertiary ophthalmology center in the northeast of Iran, between January 2013 and December 2015 were evaluated. The clinical features and risk factors of 99 (20%) consecutive patients with ocular toxoplasmosis were extracted. RESULTS: Ninety-nine including 53 (53.5%) female and 46 (46.5%) male patients with ocular toxoplasmosis were included in the analysis. Reduced vision (77%) and floaters (15.2%) were the most common presenting symptoms. The age category that was most affected by ocular toxoplasmosis was 20-40 years (range: 11-65 years) with a mean age of 27.2. All patients had retinochoroiditis, but just two had anterior uveitis. All of the extracted patients, with the exception of three patients, had unilateral involvement. None of the patients had any other medical disorders with the exception of one woman, who had diabetes. Only four recurring ocular toxoplasmosis patients were referred to the education hospital during the study. Serology data were available for just 32 patients, of which 31 (96.8%) were IgG positive, and 1 (3.2%) was IgM positive. CONCLUSION: Toxoplasma gondii was responsible for 20% of the patients of uveitis that presented to the largest ophthalmology center in the northeast of Iran. There is a high incidence of patients of ocular toxoplasmosis in the northeast of Iran, and it is a significant cause of uveitis and visual impairment in this area.

Old-world leishmaniasis presenting as a nasal mass.

Mucosal leishmaniasis is a major problem in Latin America but has been rarely noticed in our region. Although there have been a few reports of mucosal involvement especially in the oral cavity from Southwest Iran, yet none have been presented from the Northeast where Leishmania tropica is the major concern. We report a patient with endonasal leishmaniasis due to L. tropica, an extremely rare entity in immunocompetent patients in our region. He presented with a mass in the left nasal vestibule causing a sense of obstruction on the same side, accompanied by occasional rhinorrhea and mild epistaxis. This case exemplifies the need for considering leishmaniasis as a differential diagnosis for nasal obstruction in this endemic area.