mohammadbagher khadem-erfan

The role and regulation mechanisms of the interleukin-6 and 10 (IL6 and IL-10) serum levels and the interaction between CD4+ and CD8+ lymphocytes with SARS-COV-2 IgM and IgG in the context of COVID-19 infection are not fully understood.

This study was conducted on 45 COVID-19 patients and 45 healthy individuals. The IL-6 and IL-10 promoter methylation, IL-6 and IL-10 gene expression, SARS-COV-2 IgM, and IgG antibodies and CD4+ and CD8+ lymphocytes were studied by qMSP-PCR, Real-time PCR, ELISA, and flow cytometry techniques, respectively.

The male ratio and mean age of critically ill patients’ group were significantly higher in compared to controls (P< 0.05). IL-6 gene expression and serum levels were significantly increased in patients compared to controls (P=0.002, 0.001), but IL-6 promoter methylation was not significantly decreased in patients (P=0.835). The IL-10 promoter methylation and expression were not different between cases and controls (0.326, 0.455), but serum IL-10 levels were higher in patients (P< 0.001). The CD4+ and CD8+ lymphocytes decreased (P< 0.001) and mean SARS-COV-2 IgG increased (P=0.002) in the patients compared to controls.

The COVID-19 disease result in severe complications in men and elderly. The serum levels of interleukin-6 and 10 increases in COVID-19 infection, and the gene expression of these two interleukins underlying in this increase. The serum levels of IL-6, IL-10 and SARS-COV-2 IgG as well as CD4+ and CD8+ lymphocyte counts should be investigated to monitor patients and predict the course of disease.

Dental enamel formation is a complex process that is regulated by various genes. One such gene, Family With Sequence Similarity 83 Member H (Fam83h), has been identified as an essential factor for dental enamel formation. Additionally, Fam83h has been found to be potentially linked to the Wnt/β-catenin pathway.

This study aimed to investigate the effects of the Fam83h knockout gene on mineralization and formation of teeth, along with mediators of the Wnt/β-catenin pathway as a development aspect in mice.

To confirm the Fam83h-KnockOut mice, both Sanger sequencing and Western blot methods were used. then used qPCR to measure the expression levels of genes related to tooth mineralization and formation of dental root, including Fam20a, Dspp, Dmp1, Enam, Ambn, Sppl2a, Mmp20, and Wnt/β-catenin pathway mediators, in both the Fam83h-Knockout and wild-type mice at 5, 11 and 18 days of age. also the expression level of Fgf10 and mediators of the Wnt/β-catenin pathway was measured in the skin of both Knockout and wild-type mice using qPCR. A histological assessment was then performed to further investigate the results.

A significant reduction in the expression levels of Ambn, Mmp20, Dspp, and Fgf10 in the dental root of Fam83h-Knockout mice compared to their wild-type counterparts was demonstrated by our results, indicating potential disruptions in tooth development. Significant down-regulation of CK1a, CK1e, and β-catenin in the dental root of Fam83h-Knockout mice was associated with a reduction in mineralization and formation-related gene. Additionally, the skin analysis of Fam83h-Knockout mice revealed reduced levels of Fgf10, CK1a, CK1e, and β-catenin. Further histological assessment confirmed that the concurrent reduction of Fgf10 expression level and Wnt/β-catenin genes were associated with alterations in hair follicle maturation.

The concurrent reduction in the expression level of both Wnt/β-catenin mediators and mineralization-related genes, resulting in the disruption of dental mineralization and formation, was caused by the deficiency of Fam83h. Our findings suggest a cumulative effect and multi-factorial interplay between Fam83h, Wnt/Β-Catenin signaling, and dental mineralization-related genes subsequently, during the dental formation process.

Blastocystis sp. is a common intestinal protozoan that can infect humans and a wide range of animals such as mammals, birds, reptiles, and arthropods. The prevalence of Blastocystis species is relatively high in developing countries and has been reported as the most common parasite in Iran. In recent years, various studies have shown the importance of its pathogenicity in humans. Blastocystis sp. infection is associated with skin lesions and a variety of gastrointestinal disorders such as diarrhea and irritable bowel syndrome (IBS). The aim of this study was to evaluate the prevalence of Blastocystis sp. in children with cancer in Sananadaj City by PCR method.

To determine prevalence  rate of Blastocystis sp infection in children with cancer, 93 fecal samples from available cancer patients were collected from the hospitals in Sananadaj City on the basis of census method from 2019 to 2021. The samples were analyzed by molecular method (PCR) using primers in the barcoding region of 18 rRNA gene for Blastocystis sp. amplification.

The results of PCR showed 15 cases (16.1%) had 600 bp fragment for Blastocystis sp.Statistical analysis showed that Blastocystis sp.had no significant correlation with infection and residential place (Pvalue = 0. 48), age (Pvalue = 0.88) and contact to animals (Pvalue =0.83).

According to the results of this study, Blastocystis sp. is prevalent in cancer patients in Sananadaj city. These data can play an important role in the development of studies and research on Blastocystis sp.  infections in cancer patients. In general, determination of the prevalence rate of the infection and identification of the different types of Blastocystis sp.  in children with cancer and reporting these data to regional and national health officials can be beneficial in order to adopt a more appropriate treatment strategy and provide health strategies to control this infection.

Reteplase (recombinant plasminogen activator, r-PA) is a recombinant protein designed to imitate the endogenous tissue plasminogen activator and catalyze the plasmin production. It is known that the application of reteplase is limited by the complex production processes and protein’s stability challenges. Computational redesign of proteins has gained momentum in recent years, particularly as a powerful tool for improving protein stability and consequently its production efficiency. Hence, in the current study, we implemented computational approaches to improve r-PA conformational stability, which fairly correlates with protein’s resistance to proteolysis.

The current study was developed in order to evaluate the effect of amino acid substitutions on the stability of reteplase structure using molecular dynamic simulations and computational predictions.

Several web servers designed for mutation analysis were utilized to select appropriate mutations. Additionally, the experimentally reported mutation, R103S, converting wild type r-PA into non-cleavable form, was also employed. Firstly, mutant collection, consisting of 15 structures, was constructed based on the combinations of four designated mutations. Then, 3D structures were generated using MODELLER. Finally, 17 independent 20-ns molecular dynamics (MD) simulations were conducted and different analysis were performed like root-mean-square deviation (RMSD), root-mean-square fluctuations (RMSF), secondary structure analysis, number of hydrogen bonds, principal components analysis (PCA), eigenvector projection, and density analysis.

Predicted mutations successfully compensated the more flexible conformation caused by R103S substitution, so, improved conformational stability was analyzed from MD simulations. In particular, R103S/A286I/G322I indicated the best results and remarkably enhanced the protein stability.

The conformational stability conferred by these mutations will probably lead to more protection of r-PA in protease-rich environments in various recombinant systems and potentially enhance its production and expression level.

 SARS-CoV-2 virus caused the COVID-19 pandemic worldwide. One of the main pathogenic features of this virus is unpredictable clinical sequence rapidly progressing to lethal complications. Identification of clinical and laboratory prognostic factors for severe COVID-19 may contribute to the management of follow-up and treatment modalities of the disease. Complete blood count (CBC) is a fast and inexpensive laboratory test which provides a wealth of information on individual health status.

 This study aimed to evaluate the CBC test parameters in COVID-19 patients compared to healthy individuals.

 We performed a retrospective study on 147 patients with COVID-19 and 95 healthy subjects as the control group. Based on the severity of COVID-19, the patients were divided into 4 groups including mild, moderate, severe and deceased. Complete blood count parameters were recorded in all groups and obtained data were analyzed using the chi-square test at the significance level of P < 0.05 (SPSS, version 23).

 The severity of the disease increased with age and men were more likely to die from disease complications. Most CBC parameters were found to be significantly increased in patients. Significantly increased neutrophils and decreased lymphocytes were the most relevant predictors of severe disease and death. The only CBC parameter that was not associated with the disease was basophil count.

 The CBC, especially in terms of the neutrophil and lymphocyte parameters is a simple and accurate test that can be used to predict the severity of the COVID-19 disease.

Fam83h Protein is a non-secretory protein that interacts with Casein Kinase1Alpha1(CSNK1A1) through its N-terminal.

  In this study, the C-terminal domains of Fam83h in human and mouse were modeled using the I-TASER software for prediction of protein structure and function. The molecular dynamics (MD) simulations were performed by GROMACS version 5.0.2 to investigate their temporal behavior. FEL analysis was done for each model after MD. Protein-protein docking was done by PIPER to investigate the interaction of Fam83h C-terminal with cytoskeletal keratins5 as a cellular keratin model.

The results showed that the human protein is more stable and compact than the mouse protein. Assessment of the interaction between the C-terminal of the mouse Fam83h and Keratin-5 proteins showed the residues Arg378, Pro391, Ser663, Glu710, Arg923 in mouse Fam83h protein made hydrogen bonds with Leu474, Arg417, Tyr453, Glu 397, Gln 396 and Glu 390 of the chain A and B of mouse keratin-5, respectively. Human Fam83h and keratin 5 form hydrogen bonds via residues of Thr433, His447 and Arg477 and Leu473, Gly476, Glu420 and Arg407.

According to the previous experimental reports, Fam83h can interact with keratin cytoskeleton and has a role in cancer progression. It can be concluded that Fam83h protein can directly interact with keratin filaments through its C-terminal. Therefore, our hypothesis based on  in-silico study is: non-sense mutation in C-terminal of Fam83h protein may lead to truncated protein and subsequently disruption of keratin cytoskeleton bundling which can be regarded as a mechanism involved in cancer progression.

It is important to find novel therapeutic molecular targets for curing Parkinson’s disease (PD). Accordingly, this study aimed to evaluate the effect of over-expression of the survivin gene, a gene frequently reported as neuroprotective, on the in vitro model of PD.  Survivin was over-expressed in SH-SY5Y cells. Next, the cells were treated with rotenone (500 nM) for 24 hr. Then, viability and the total antioxidant capacity were assessed. The expression levels of 15 important genes of key cellular processes (oxidative stress, apoptosis, cell cycle, and autophagy) were assessed. The studied genes included survivin, superoxide dismutase, catalase, BAX, bcl2, caspase 3, caspase 8, caspase 9, p53, SMAC, β-catenin, mTOR, AMPK, ATG7, RPS18. The apoptosis level and the frequency of cell cycle stages were assessed by flow cytometry. For analyzing the data, the ANOVA test followed by Tukey’s test was used to evaluate the significant differences between the experimental groups. P<0.05 was considered significant.  Survivin could significantly decrease the rotenone-induced apoptosis in SH-SY5Y cells. The rotenone treatment led to down-regulation of catalase and up-regulation of bax, bcl2, caspase 3, caspase 8, P53, β-catenin, and ATG7. Survivin could significantly neutralize the effect of rotenone in most the genes. It could also increase the total antioxidant capacity of SH-SY5Y cells.  Survivin could prevent the toxic effect of rotenone on SH-SY5Y cells during the development of in vitro PD model via regulating the genes of key cellular processes, including anti-oxidation, apoptosis, cell cycle, and autophagy.

Engyodontium album, Fusarium venenatum, and Phanerochaete chrysosporium fungi have an important role in the production of proteinase K, Quorn mycoprotein, and bioremediation, respectively. There are several techniques for the detection of fungi in soil. The purpose of this study was to detect and identify E. album, F. venenatum, and P. chrysosporium, directly from soil using the Polymerase Chain Reaction (PCR) method.

A total of 240 soil samples were collected from different regions of Iran, including Tehran, Zanjan, Hamadan, Kermanshah, Kurdistan, and Hormozgan Provinces. The DNA was extracted and purified directly from soil samples with the modified phenol-chloroform method and by PVPP (Polyvinylpolypyrrolidone) column, respectively. The PCR method was performed using designed specific primers. The PCR products of the E. album, F. venenatum and P. chrysosporium with an approximate size of 248, 202, and 502 bp were sequenced, respectively.

In this study two isolates of P. chrysosporium, 1 isolate of F. venenatum, and 1 isolate of E. album were identified. The fungi detected with specific primers in soil samples were compatible with the results of sequencing.

This investigation described a reliable method that can be used to detect important fungi in the industries and biotechnology directly in soil using specific primers. The results can provide an appropriate platform for next applied research and mass production of valuable fungal products in industries and biotechnology.

Mouse embryo culture condition is an essential part of transgenic, reproductive and developmental biology laboratories. Mouse embryonic culture media may have a high risk of serum contamination with pathogens.

To investigated the effect of sericin as an embryo culture medium supplement on in vitro maturation (IVM), in vitro fertilization (IVF), and development of the preimplantation embryo in mice.

The effects of sericin at three concentrations (subgroups) of 0.1%, 0.5%, and 1% as a medium supplement on IVM, IVF, and in vitro development of mouse embryos were separately investigated and compared with a sericin-free (control) group. The cumulative effect of the three concentrations was evaluated for IVM+ in vitro development and IVF+ in vitro development as follow-up groups.

In the IVM group, compared to the control group, the number of oocysts reaching the MII stage was significantly higher when 1% sericin was used (161/208 = 77.4%). No significant results were observed in the IVF and in vitro development groups with different concentrations of sericin compared to the control group. Among the follow-up groups, in the IVM + in vitro development group, the number of oocytes was higher after passing the IVM and IVF and reaching the blastocysts stage when 1% sericin was used, compared with other sericin subgroups. A significant difference was also noted when compared with the control group (p = 0.048). The IVF+ in vitro development study group, on the other hand, did not show any significant relationship.

It can be concluded that 1% sericin can be used as a supplement in mouse embryo cultures to improve the IVM rate. Also, based on the findings, sericin appears to be an effective supplement which can have a positive effect on the development of embryos derived from IVM.

Toxocariasis is a disease caused by Toxocara nematodes and occurs from consuming their eggs. The main hosts of these worms are dogs and cats. The disease in humans becomes a visceral larva migrans (VLM). This descriptive cross-sectional study was conducted to determine the prevalence of toxocariasis in children aged 6–14 years.

This cross-sectional descriptive study was conducted from Jun 1 2016 to Dec 1 2017 in Sanandaj, west of Iran. A total of 182 serum samples were collected from children age 6 to14 yr referred to medical diagnostic laboratories. Demographic data (age, sex, and parents' literacy status), clinical signs (cough, headache, fever, abdominal pain), and the history of contact with dogs and cats was collected by a questionnaire. The presence of anti-Toxocara IgG antibody was detected by T. canis IgG ELISA (IBL, Germany) kit.

Of 182 subjects, 97 (53.3%) were male and 85 (46.7%) female. The average age was 9.2 years. Antibodies against T. canis were positive in three cases (1.65%) of all the studied subjects.

The results showed a low prevalence of toxocariasis in children studied.

Toxoplasma gondii is a widely-distributed parasite all over the world whose attribut-ed severe afflicting complications in human necessitate the development of serodiagnostic tests and vaccines for it. Immunological responses to monovalent vaccines and the application of diagnostic reagents including single antigens are not optimally effective. Bioinformatics approaches were used to introduce these epitopes, predict their immunogenicity and preliminarily evaluate their potential as an effective DNA vaccine and for serodiagnostic goals.

A 3D structure of proteins was predicted by I-TASSER server, and linear and conformational B cell and T cell epitopes were predicted using the online servers. Then, the predicted epitopes were constructed and called Toxoeb, and their expression in the prokaryotic and eukaryotic cells was demonstrated using SDS-PAGE. In the next step, Western blotting with pooled sera of mice infected with T. gondii was done.

The current in silico analysis revealed that the B cell epitopes with high immunogenicity for GRA4 protein were located in the residues 34-71, and 230-266, for GRA14 in 308-387, for SAG1 in 182-195, 261-278, and for GRA7 in residues 101-120, 160-176. The T cell epitopes were selected in overlapping regions with the B cell epitopes. The immunogenic region for GRA4 are in the residues 245-253, 50-58, and 40-54, for GRA14 in 307-315, 351-359, and 308-322, for SAG1 261-269, and 259-267, and for GRA7 in the residues 103-112, and 167-175. The results of the western blotting showed that the expressed protein had immunogenicity.

Our constructed multi-epitope of T. gondii could be considered as a candidate for diagnostic and vaccination purposes.

 Multiple sclerosis (MS) is one of the chronic inflammatory diseases of the nervous system. The cause of the disease has not yet been clearly identified. Environmental factors and infections, including the toxoplasma, are hypothesized to be the cause of the disease. Toxoplasma has important effects in diseases related to the nervous system. Our goal was to compare the serum antibody level against toxoplasma in patients with MS and healthy people in Sanandaj, Iran.     In this case-control study, 100 patients with MS who were registered in the MS Society of Sanandaj and 200 matched healthy blood donors from the Sanandaj Blood Transfusion Organization (control group) were studied from 2015 to 2016. 5 ml blood sample was obtained from all subjects and then after isolation of patients' sera, IgG antibodies against toxoplasma -antigens were measured by ELISA method. Data were analyzed by SPSS software and Chi-square test.     Toxoplasma antigen was found in 13% of patient group (p=0.204) and 14.5% of control group, there is no significant difference between the levels of anti-toxoplasma antibody in the 2 groups (P > 0.05). The age of patients was between 20-40 years and the control group was also matched accordingly. The place of residence of all patients and all the control group was urban.     It seems that toxoplasma gondii has not relation with MS.

Iran is among the first six countries in the world with the highest annual incidence of cutaneous leishmaniasis. This study aimed at estimating the incidence, burden, and trend of cutaneous leishmaniasis at the national level in Iran from 1977 to 2015. This study was conducted in 2017, used Disability Adjusted Life Years (DALYs) index, recom-mended by the WHO for assessing the Global Burden of Diseases, to estimate the burden of cutaneous leish-maniasis. The data on the incidence of the disease and the number of cases was obtained from the communi-cable diseases surveillance system (Center for Communicable Diseases Control (CCDC), Ministry of Health and Medical Education (MOHME) of Iran) that routinely collects data from all over the country; in addition, some data was extracted by reviewing the texts. Considering the views expressed in a panel of experts, to calcu-late the actual incidence of the disease, the number of registered cases was multiplied by 5. The incidence of cutaneous leishmaniasis varied from approximately 50 to 250 cases per 100,000 population during the studied period. During these years, the incidence of the disease was higher in males than females. Moreover, the burden of cutaneous leishmaniasis varied between 1.18 and 5.7 DALYs per 100000 population during the studied period. The incidence and burden of cutaneous leishmaniasis have not signifi-cantly decreased in recent years. Despite the implementation of a program for controlling cutaneous leishmaniasis in Iran since 1977, the incidence and burden of the disease are still high in the country; it is seriously alarming for policy makers and managers of the health system in Iran, indicating the presence of some problems in controlling the disease.

Calodium hepaticum (syn. Capillaria hepatica) is a nematode with worldwide distribution among rodents (Rattus rattus, Rattus norvegicus) and also can infect human beings through ingestion of embryonated eggs which causes zoonosis Capillariasis. This study aimed to determine the Capillaria hepatica infection in Rattus spp. in Sanandaj, Kurdistan province of Iran, as well as orally infecting Balb/c mice with embryonated eggs.
Material and Area of study was the city of Sanandaj located in the west of Iran, in which four Rattus norvegicus, six Rattus rattus and 10 Mus musculus were captured using live traps in a period of more than two years. Livers were examined for Capillaria hepatica infection, and sectioning of the livers and staining with Hematoxylin and Eosin were done. Embryonated eggs were used in order to experimentally infect three Balb/c mice orally.
Capillaria hepatica was detected in five out of 20 (25%) livers; rates of infection were 50% (2/4) for Rattus norvegicus, 50% (3/6) for Rattus rattus, and 0% (0/10) for Mus musculus. Cross sections of the livers showed active infections as eggs were deposited beside the worms, and they were also observed inside the female worms. After performing the autopsy on the three mice that had been infected experimentally, there was no sign of infection based on gross and microscopic examinations.
Discussion and This study is the first report on Capillaria hepatica infection in Rattus spp. in Sanandaj. The main reservoir hosts of Capillaria hepatica infection in different areas of Sanandaj were Rattus rattus and Rattus norvegicus; on the other hand, no significant role could be considered for either infection or transmission of Capillaria for Mus musculus in these areas.

The effect of frequent examinations on the students’ learning has had inconsistent results. This study aimed to assess the effectiveness of fre­quent announced quizzes on the learning of a representative sample of Iranian medical students. This experimental study was conducted among 37 fifth semester medi­cal students who had taken the course in Protozoology and Helminthology, in which the same basic information were provided about different types of proto­zoa and worms. Initially, in the teaching of helminthology, ten routine ses­sions were handled with lectures and interactive questions and answers. Then at the beginning of the protozoology topic in the beginning of all of the next 9 ses­sions, the students were informed that they will have a quiz at the end of each session. At the end of the semester, the total scores of quizzes were compared with the mean final scores of protozoology and helminthology using paired t and repeated measure tests. The mean final scores of the protozoology lesson were not significantly different from that of the helminthology (10.45 ± 2.75 vs.11.25 ± 2.56 on the scale of 20, respectively, P=0.13). There was no significant difference in the mean score of the five quizzes compared with the mean final term score of protozo­ology. The overall mean scores in the helminthology lesson (11.25±2.56), protozoology lesson (10.45±2.75), and the quizzes (9.16 ± 3.55) were significantly different (P <0.0001). Frequent announced quizzes were not effective on increasing the medical students'' motivation and learning.

Pentavalent antimonials are the first line drugs for the treatment of leishmaniasis. Unresponsiveness of Leishmania spp. to antimonial drugs is a serious problem in some endemic areas. Investigations on molecular mechanisms involved in drug resistance are essential for monitoring and managing of the disease. Calcineu­rin is an essential protein phosphatase for number of signal transduction pathways in eukaryotic cells and it has a mediated role in apoptosis. This study aimed to determine of biomarker(s) in Glucantime® resiatance strain of L. infan­tum. We used cDNA amplified fragment length polymorphism (cDNA-AFLP) and real time-RT PCR assays to compare gene expression profiles at the mRNA levels in resistant and susceptible L. infantum field isolates. The cDNA-AFLP results showed downlegulation of calcineurin in resis­tant isolate in comparison with susceptible one. Significant downregulation of calcineu­rin (0.42 fold) (P<0.05) was found in resistant isolate compared to suscepti­ble one by Real time-RT PCR. This is the first report of calcineurin implication in Glucantime® drug resistance of field (natural) isolate of L. infantum. Downregulation of calcineurin could protect parasites from antimonial-induced apoptosis.