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Background: We aimed to investigate the molecular genotyping of Giardia duodenalis in Humans in the Yazd County, Central, Iran. Methods: Total of 35 fecal samples were collected from patients referred to Yazd Central Laboratory, Yazd, Iran from February to July 2022. All the samples were included in this study after microscopic observation of G. duodenalis. DNA samples were extracted using related kit and were analyzed by Nano Drop. The molecular assessment was carried out using semi-nested PCR using the target gene of gdh. All amplified samples were sequenced using Sanger method. BLAST analyzed the sequences for assemblage identification. Results: Out of 35 samples, 24 (68.57%) and 11 (31.43%) were male and female, respectively. All included samples were amplified using the specific gdh primer pair. The molecular analysis showed 17 isolates (48.57%) as assemblage BIV, 8 isolates (22.86%) as assemblage BIII, 6 isolates (17.14%) as assemblage AII and 4 isolates (11.43%) as assemblage AIII (P<0.05). Conclusion: Assemblages A and B are the most prevalent in Central Iran. The molecular identification of G. duodenalis isolates from animals and implementing control programs. [ABSTRACT FROM AUTHOR]

Background and Aim: Toxoplasmosis is a common parasitic infection that can endanger mother's and neonates' health during pregnancy. The disease is also prevalent in Iran. This study intended to evaluate the seroepidemiology of toxoplasmosis in neonates and postpartum mothers referred to health centers of Yazd in Iran in 2020. Materials and Methods: Totally, 184 postpartum mothers and 184 neonatal umbilical cords in health centers of Yazd were evaluated for Toxoplasma infection through a specific IgM and IgG antibodies kit. The obtained data were analyzed by SPSS18. Results & Conclusion: Out of 184 samples of postpartum mothers, 8 cases (4.35%) were seropositive, and 176 (95.65%) were seronegative for IgG antibody; moreover, 7 cases (3.80%) were seropositive, and 177 (96.20%) seronegative for IgM antibody. Also, 184 neonatal umbilical cords were IgM negative, and no toxoplasmosis infection was reported. No significant correlation was found between seroprevalence of Toxoplasma infection and caring for pets, consumption of raw meat, level of education, blood type, job, living area and type of delivery (P>0.05). However, a significant correlation was identified between the number of deliveries and the prevalence of toxoplasmosis (P=0.014). This study also illustrated a low prevalence of Toxoplasma infection in postpartum mothers and no congenital transmission of the disease in diverse health centers of the province. However, there was no statistically significant relationship between risk factors and the prevalence of Toxoplasma. [ABSTRACT FROM AUTHOR]

Genetically Modified (GM) foods are among the most important achievements of biotechnology. Given the safety importance of transgenic rice, this study was conducted to investigate the effect of GM rice consumption on serum concentrations of tumor markers in rats. In this experimental intervention, we used the blood serum samples from the Biobank taken from 60 males and 60 female Sprague-Dawley (SD) rats fed on three different diets, including rat's standard food, non-GM rice, and GM rice after 90 day. Tumor markers including Carcinogenic embryonic antigen (CEA), Alpha-Fito protein (AFP), Cancer Antigen 19-9 (CA19-9), Cancer Antigen 125 (CA125), and Cancer Antigen15-3 (CA15-3) were assessed by enzyme-linked immune sorbent assay (ELISA) method. Statistical analysis was conducted via SPSS software. The results show that the concentrations of tumor markers were within the normal range in the SD rats treated with diet, and since the concentration of tumor markers was lower than the acceptable index determined, according to the kit standard in both groups, no obvious carcinogenic effect was found. However, these findings are not enough to make a final decision regarding the safety assessment of GM rice consumption. [ABSTRACT FROM AUTHOR]

Objective: Male genital tract infections have been associated with infertility, and Escherichia coli has drawn increasing attention as an important bacterium in this context. This investigation aimed to characterize and compare the distributions of O-antigen serogroups of E. coli in the semen samples of fertile and infertile men. Methods: In this case-control study, semen samples were collected from 618 fertile and 1,535 infertile men. The E. coli-positive samples were evaluated in terms of concentration, morphology, viability, and motility parameters according to the World Health Organization 2010 guidelines. Finally, different serogroups of E. coli were identified by multiplex polymerase chain reaction targeting the O-antigen variations of the bacterium. Results: The prevalence of E. coli among fertile men was significantly higher than among infertile men (p<0.001). The sperm morphology, viability, and motility in the E. coli-positive fertile group were significantly higher than in the E. coli-positive infertile group (p<0.001). E. coli O6 was the most prevalent serogroup found in both groups. However, there was no significant difference in the frequency of different serogroups of E. coil between the two groups (p=0.55). Conclusion: Despite the higher prevalence of E. coli among fertile men, E. coli had more detrimental effects on semen parameters in infertile men. There was no significant difference in E. coli serogroups between the fertile and infertile groups. [ABSTRACT FROM AUTHOR]

Background: Toxoplasma gondii, an obligate intracellular protozoan, causes toxoplasmosis. The aim of this study was molecular detection of T. gondii in breast and goat milk samples by the molecular method in the central Iran. Methods: Totally, 300 human' and 200 goats' milk samples were collected randomly from different regions of central Iran in 2018. DNA extraction was performed by the salting-out method. Molecular detection of the parasite was done by nested-PCR using the specific primer pairs. Statistical analysis was performed by SPSS 23 using descriptive and Chi-square tests. Results: Out of 300 human milk samples, 1 sample (0.3%) was infected with T. gondii. Out of 200 samples of goat milk, 11 samples (5.5%) showed infection with T. gondii. The frequency of infection in goat's milk samples was 4.36% in the south and west, 1.9% in northern regions, and 2% in eastern regions of the province. The statistical analysis showed no significant difference between toxoplasmosis and different geographical regions in the province. Conclusion: Because of the popularity of the goat milk and the transfection probability with the milk to humans, it is recommended to boil milk prior to use. Furthermore, case contamination of T. gondii in the human milk sample showed one of the important paths for infection transmission, which requires further studies. [ABSTRACT FROM AUTHOR]

Background: Leishmaniasis is a prevalent tropical disease caused by more than 20 Leishmania species (Protozoa, Kinetoplastida and Trypanosomatidae). Among different clinical forms of the disease, cutaneous leishmaniasis is the most common form, with an annual 0.6-1 million new cases reported worldwide. This disease's standard treatment is pentavalent antimonial (SbV) that have been used successfully since the first half of the 20th century as a first-line drug. However, treatment failure is an increasing problem that is persistently reported from endemic areas. It is important to define and standardize tests for drug resistance in cutaneous leishmaniasis. SbV must be reduced to its trivalent active form (SbIII). This reduction occurs within the host macrophage, and the resultant SbIIIenters amastigotes via the aquaglyceroporin1 (AQP1) membrane carrier. Overexpression of AQP1 results in hypersensitivity of the parasites to SbIII, but resistant phenotypes accompany reduced expression, inactivation mutations, or deletion of AQP1. Hence, in this study, a phylogenetic analysis using barcode gene COXII and kDNA minicircle and expression analysis of AQP1 were performed in treatment failure isolates to assess the isolates' molecular characteristics and to verify possible association with drug response. Methods: Samples in this study were collected from patients with cutaneous leishmaniasis referred to the Diagnosis Laboratory Center in Isfahan Province, Iran, from October 2017 to December 2019. Among them, five isolates (code numbers 1-5) were categorized as treatment failures. The PCR amplification of barcode gene COXII and kDNA minicircle were done and subsequently analyzed using MEGA (10.0.5) to perform phylogenetics analysis of Treatment failures (TF) and Treatment response (TR) samples. Relative quantification of the AQP1 gene expression of TF and TR samples was assessed by real-time PCR. Results: All samples were classified as L. major. No amplification failure was observed in the cases of barcode gene COXII and kDNA minicircle amplification. Having excluded the sequences with complete homology using maximum parsimony with the Bootstrap 500 method, four major groups were detected to perform phylogenetic analysis using COXII. The phylogenetic analysis using the barcode target of minicircle showed that all five treatment failure isolates were grouped in a separate sub-clade. Conclusions: We concluded that the barcode gene COXII and the minicircle kDNA were suitable for identification, differentiation and phylogenetic analysis in treatment failure clinical isolates of Leishmania major. Also, AQP1 gene expression analyses showed that treatment failure isolates had less expression than TR isolates. The isolate with TF and overexpression of the AQP1 gene of other molecular mechanisms such as overexpression of ATP-binding cassette may be involved in the TR, such as overexpression of ATP-binding cassette which requires further research. [ABSTRACT FROM AUTHOR]

Background and Objectives: Due to the important role of Streptococcus agalactiae, Group B streptococci (GBS), in production of invasive disease in neonates, investigation regarding the pathogenicity and antibiotic resistance factors is necessary in selecting the appropriate therapeutic agents. Beside capsule, the pilus has been currently recognized as an important factor in enhancing the pathogenicity of GBS. Resistance of GBS to selected antibiotics is noticeably increasing which is mainly due to the anomalous use of these drugs for treatment. The aim of this study was to determine the prevalence of pili genes followed by antibiotic susceptibility of GBS, previously serotyped, isolated from pregnant women in the city of Yazd, Iran. Materials and Methods: Fifty seven GBS from pregnant women were subjected to multiplex PCR for determination of PI-1, PI-2a and PI-2b pilus-islands and simultaneously, the phenotype of antibiotic resistance to penicillin, tetracycline, erythromycin, clindamycin, gentamycin and levofloxacin was determined. Antibiotic resistance genes (ermA, ermB, mefA, tetM, int-Tn) were further diagnosed using PCR and multiplex PCR. Results: PI-1+PI-2a with 71.9%; followed by PI-2a (21.1%) and PI-2b (7%) were observed. PI-1+PI-2a in serotype III was (73.2%), serotype II, Ia, Ib and V were 12.2%, 9.8%, 2.4% and 2.4% respectively. GBS penicillin sensitive was 89.5% and 96.5% resistance to tetracycline. The frequency of resistance genes were as follows: tetM (93%), ermA (33.3%), ermB (8.8%), int-Tn (80.7%) and mefA (0). Conclusion: Majority of GBS contained PI-1+PI-2a. Hence presence of this pilus stabilizes the colonization, therefore designing a program for diagnosing and treatment of infected pregnant women seems to be necessary. [ABSTRACT FROM AUTHOR]

Background and Objectives: Burn wound infections have emerged as an important cause of morbidity and mortality in patients due to prolonged hospital stay. Pseudomonas aeruginosa, is the second cause of bacterial burn wound infections. Resistance mechanisms among P. aeruginosa are intrinsic or acquired. Intrinsic resistance mechanisms among P. aeruginosa isolates are inducible AmpC cephalosporinase, decrease of specific porin OprD, and overexpression of RND efflux pump. The aim of this study was detection of mutations in nalC gene in carbapenem resistant P. aeruginosa isolated from burn wounds. Materials and Methods: In this cross-sectional study, 180 burn-wound specimens were collected. Suspected lactose-negative colonies were identified by conventional biochemical methods. Kirby-Bauer and Etest methods were used for susceptibility testing. PCR and sequencing techniques were used for the detection of nalC mutation. Results: Out of 180 specimens received in the laboratory, 54 of isolates were isolated and identified as P. aeroginosa (30%). Of these isolates 20 (37%) were resistant to at least two carbapenems simultaneously. From these carbapenem resistant isolates, 19 (95%), 14 (70%), 14 (70%), 19 (95%) and 16 (80%) were resistant to imipenem, cefepime, piperacillin, ceftizoxime and gentamicin, respectively. Only 1 (2%) isolate was sensitive to all carbapenems and did not has mutation in nalC gene, 20 (37%) isolates were resistant to at least two carbapenems, and had mutations in nalC gene (Gly71?Glu and Ser209?Arg). Conclusion: As the results showed, mutation in efflux pump was observed in carbapenem resistant isolate and this confirmed that the indiscriminate use of antibiotics for treatment or prophylaxis can increase mutation in efflux pump. [ABSTRACT FROM AUTHOR]

Background: The first line treatment for cutaneous leishmaniasis is pentavalent antimony such as sodium stibogluconate (pentostam) and meglumine antimonite (glucantime). One of the most important ways to uptake the drug is by a trans-membrane protein, called aquaglyceroporin encoded by Aquaglyceroprotein1 (LmAQP1). In this study, molecular characterization of LmAQP1 was reported. Methods: Leishmania major (MRHO/IR/75/ER) promastigotes were cultured, and then DNA extraction and RNA extraction were done and followed by cDNA synthesis. Amplicons resulted from PCR and RT-PCR using specific primers were purified and sequenced. Molecular characterization was done by bioinformatically software such as BLST, ClustalW2, and RMSD. Results: Amplicons resulted from PCR and RT-PCR showed equal size in length. BLASTn analysis showed a point nucleotide change in LmAQP1 gene that encoded 282-amino-acid long protein with a mutation at position 154 including replacement of alanine by threonine. The observed mutation in the interested gene was assessed using the above-mentioned software. The mentioned gene was submitted at GenBank, NCBI with accession number of KU514052. Conclusion: The functional prediction of the protein encoded from LmAQP1 showed that the mentioned mutation could not affect the three-dimension structure, but it may modify the drug uptake potential of this important channel. Based on from LmAQP1 role, it seems to be an appropriate candidate for drug development. According to search through internet, this is the first report of LmAQP1 from L. major (MRHO/IR/75/ER). [ABSTRACT FROM AUTHOR]

Background: Regarding the antimonial-resistant of Leishmania spp., understanding of related mechanism is necessary. One of the most important involved molecules is aquaglyceropin1 (AQP1). The aim of this study was molecular analysis of AQP1 gene from antimonial-resistant clinical isolates and its expression. Methods: Overall, 150 patients with cutaneous leishmaniasis referring to the reference laboratories of Yazd and Varzaneh,, located 105km southeast of Isfahan and 240km away from Yazd, were assessed from Jun 2015 to Dec 2017. After sampling, staining was done and evaluated for Leishman by microscope. Samples were collected in RNAlater solution for gene expression analysis in non-healing isolates. DNA extraction was performed from each slide with Leishman body. All patients with L. major isolates detected by ITS1-PCR-RFLP were followed for finding the resistant isolates, consequence of molecular characterization of AQP1 using PCR-RFLP. Gene expression of AQP1 from all resistant isolates was assessed in comparison with the one in a sensitive isolate. Statistical analysis was done using SPSS. The significance level was considered ≤0.05. Results: Five isolates were detected as antimonial resistant. Molecular detection and identification were appeared that all were L. major. The molecular characterization of AQP1 showed G562A mutation. Gene expression of AQP1 in resistant isolates showed 1.67 fold higher than the sensitive isolate. Conclusion: We reported a new point mutation of G562A in AQP1 gene involved in molecular mechanism in resistant isolates. [ABSTRACT FROM AUTHOR]

Microorganisms used as a biodegradation technique can provide economic benefits and can be compatible with the environment. The aim of this study was to isolate low-density polyethylene (LDPE) degrading bacteria. The waste samples were collected from Yazd landfills. Biochemical and molecular tests based on 16S rDNA sequencing were done to identify the superior isolates. Biodegradation rate was measured using weight loss measurement, spectroscopic analysis with Fourier-transform infrared (FTIR) spectroscopy and an investigation of surface morphology changes using scanning electron microscopy (SEM). Two biodegrading isolates were identified as Pseudomonas aeruginosa strain SKN1 and strain SKN2. The weight loss of LDPE strips was measured at 10.32%. SEM micrographs showed the surface degradation and colony formation on LDPE strips. The FTIR spectrum revealed the structural changes on LDPE strip surfaces. Bio-decomposition was observed to have taken place and the bacterial strains had a special ability to biodegrade LDPE wastes. [ABSTRACT FROM AUTHOR]

Background and Objectives: Antibiotic resistance in Pseudomonas aeruginosa is an increasing health problem. Integrons are associated with a variety of gene cassettes, which confer resistance to multiple classes of antibiotics. This study aimed at screening the presence of class 1, 2 and 3 integrons in P. aeruginosa in Yazd, Iran. Materials and Methods: This study was carried out on P. aeruginosa strains from March 2016 to March 2017. Clinical specimens were initially identified by the standard biochemical methods and their resistance patterns to antibiotics were studied using the disc diffusion method. PCR was carried out for the detection of class 1, 2 and 3 integrons using intI1, intI2 and intI3 gene primers, respectively. Results: Antimicrobial susceptibility test showed that 75% of isolates were detected as multi-drug resistant (MDR), and lowest resistance was observed in ciprofloxacin (48.6%) and most resistance was in gentamicin (63.2%). Moreover, PCR results showed that 22 (15.3%) and 119 (82.6%) of P. aeruginosa isolates carried intI2 and intI1 genes, but intI3 gene was not found. Conclusion: Since it is customary to observe Class I integrons in P. aeruginosa isolated from clinical samples, they are often responsible for antibiotic resistance gene transfer, which calls for evaluation of integrons as contributing factors in antibiotic resistance. [ABSTRACT FROM AUTHOR]

Background: Klebsiella pneumoniae (K. pneumoniae) is an opportunistic pathogen that could be resistant to many antimicrobial agents. Resistance genes can be carried among gram-negative bacteria by integrons. Enzymatic inactivation is the most important mechanism of resistance to aminoglycosides. In this study, the frequencies of two important resistance gene aac(6')-IIa and ant(2'')-I, and genes coding integrase I and II, in K. pneumoniae isolates resistant to aminoglycosides were evaluated. Methods: In this cross-sectional study, an attempt was made to assess the antibiotic susceptibility of 130 K. pneumoniae isolates obtained from different samples of patients hospitalized in training hospitals of Yazd evaluated by disk diffusion method. The frequencies of aac(6')-IIa, ant(2'')-I, intl1, and intl2 genes were determined by PCR method. Data were analyzed by chi-square method using SPSS software (Ver. 16). Results: our results showed that resistance to gentamicin, tobramycin, kanamycin, and amikacin were 34.6, 33.8, 43.8, and 14.6%, respectively. The frequencies of aac (6')-IIa, ant(2'')-I, intl1, and intl2 genes were 44.6, 27.7, 90, and 0%, respectively. Conclusion: This study showed there are high frequencies of genes coding aminoglycosides resistance in K. pneumoniae isolates. Hence, it is very important to monitor and inhibit the spread of antibiotic resistance genes. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is an important opportunistic pathogen that mainly infects critically patients in intensive care units (ICU). The production of plasmid-mediated extendedspectrum β-lactamases (ESBLs) is one of the most important mechanisms of resistance against β-lactam antibiotics. This study aimed to evaluate the prevalence of ESBLs in A. baumannii isolated from ICU of Ghaem hospital, Mashhad, Iran. A total of 140 A. baumannii isolates recovered from hospitalized patients in ICU of Ghaem hospital in Mashhad city from December 2014 to March 2015. Identification of A. baumannii isolates carried out using biochemical laboratory methods and then confirmed by OXA-51 PCR screening. Susceptibility testing performed using disk diffusion (Kirby-Bauer) method as recommended by CLSI guidelines. A. baumannii isolates screened for production of ESBLs using combination disk test. blaPER, blaGES, blaTEM, blaSHV, blaCTX, blaVEB and blaOXA-10 beta-lactamase genes detected using conventional PCR. The most antibacterial resistance was against cefuroxime (99.3%) and colistin was the most effective antibiotic. None of the isolates were ESBL producer by combination disk test. However, results of PCR revealed that the prevalence of blaPER, blaGES and blaTEM genes were 7.1%, 4.3% and 27.1%, respectively. blaCTX, blaVEB, and blaOXA-10 were not found in any of isolates. According to the results, the high resistance was seen against selected antibiotics and the phenotypic tests are not sufficient alone for determination of ESBLs producer of A. baumannii isolates. So, molecular tests are also necessary for detection of these enzymes. [ABSTRACT FROM AUTHOR]

Background: One of the most important items in molecular characterization of food-borne pathogens is high quality genomic DNA. In this study, we investigated three protocols and compared their simplicity, duration and costs for extracting genomic DNA from Linguatula serrata. Methods: The larvae were collected from the sheep's visceral organs from the Yazd Slaughterhouse during May 2013. DNA extraction was done in three different methods, including commercial DNA extraction kit, Phenol Chloroform Isoamylalcohol (PCI), and salting out. Extracted DNA in each method was assessed for quantity and quality using spectrophotometery and agarose gel electrophoresis, respectively. Results: The less duration was regarding to commercial DNA extraction kit and then salting out protocol. The cost benefit one was salting out and then PCI method. The best quantity was regarding to PCI with 72.20±29.20 ng/µl, and purity of OD260/OD280 in 1.76±0.947. Agarose gel electrophoresis for assessing the quality found all the same. Conclusion: Salting out is introduced as the best method for DNA extraction from L. seratta as a food-borne pathogen with the least costand appropriate purity. Although, the best purity was regarding to PCI but PCI is not safe as salting out. In addition, the duration of salting out was less than PCI. The least duration was seen in commercial DNA extraction kit, but it is expensive and therefore is not recommended for developing countries where consumption of offal is common. [ABSTRACT FROM AUTHOR]

Background: Y-chromosome short tandem repeats (Y-STRs) are genetic markers with practical applications in human identification and population studies. Aim: Here we present the allelic and haplotype frequencies of 8 Y-STR loci most commonly used in forensic medicine in 103 unrelated native males of Isfahan province, central part of Iran. Subjects and methods: The cases were selected on the basis of strict criteria to assure pure native populations of Isfahan origin. DNA extracted from peripheral blood samples and PCR amplified for each marker. Y-specific STR loci DYS19, DYS385, DYS389I, DYS389II, DYS390, DYS391, DYS392 and DYS393 were included in this study. Results: The most common alleles for each locus were: DYS19, allele 12; DYS385, allele 12; DYS389I, allele 13; DYS389II, allele 29; DYS390, allele 24; DYS391, allele 10; DYS392, allele 11; and DYS393, allele 13. Gene diversity value was calculated from the allelic frequency for each locus. The average gene diversity was 0.6518. A total of 101 haplotypes were observed in eight Y-specific STR loci, the haplotype diversity was raised to 0.986. Conclusion: The results revealed that a set of eight Y-specific STR loci were able to discriminate most of the male individuals in the population studied. A search through the Y Haplotype Reference Database demonstrated 21 matched haplotypes to 160,693 haplotypes, exclusively with Eurasian-European, Eurasian, and Eurasian-Indo Iranian populations. [ABSTRACT FROM AUTHOR]

Background: Genotyping of Crithidia species is critical in order to design appropriate programs for control and prevention of cutaneous leishmaniasis (CL). Objectives: The purpose of this project was study and investigation genetic polymorphism of Crithidia in Isfahan city. Patients and Methods: In this descriptive-analytical study, 602 samples were isolated from suspicious patients with cutaneous leishmaniasis who referred to Leishmaniasis Research Center or other health centers in the vicinity of Isfahan. After culturing of the samples in NNN (Novy MacNeal Nicole, ©Merck) medium, DNA extraction was performed. Molecular detection and identification was done using PCR-RFLP technique. Results: Results showed that 50 samples (8.31%) were negative. Out of 201 samples (33.39%) were Crithidia Spp which 27 cases (13.44%) were Crithidia fasciculata, 11 cases (5.47%) were Crithidia luciliae and the rest (163 cases or 81.9%) were similar to Trypanosomatidae which submitted at gen bank with accession number of GQ331988. BLAST showed that the last ones have 92% and 89% similarity to C. fasciculata and C. luciliae, respectively. Out of 351 patients (58.31%) identified as Leishmania spp. Conclusions: The results of the present study suggested that there is some recombinant species in Isfahan and surrounding area that could be an important agent of cutaneous leishmaniasis. [ABSTRACT FROM AUTHOR]

The aim of the study was to determine the role of transforming growth factor-beta1 (TGF-β1) and Foxp3 (rs3761548) promoter polymorphisms in Iranian women with recurrent spontaneous abortion (RSA). Eighty women with RSA were compared with eighty in a control group. Serum levels of TGF-β1 were measured using ELISA and Foxp3 (rs3761548) promoter polymorphisms using a PCR-RFLP technique. In addition, serum levels of TGF-β1 were compared in different genotypes in the two groups. The women's ages in the two groups were similar (30.15 ± 4.42 years [RSA] vs. 29.97 ± 4.51 [control]) as were serum TGF-β1 concentrations in case and control groups (53.42 ± 2.08 ng/ml in control and 56.31 ± 2.58 ng/ml in the RSA group; p = 0.4). Furthermore, there was no significant difference in the genotype frequencies of the rs3761548 Foxp3 gene between the two groups ( p = 0.3) and the levels of TGF-β1 were similar in different genotypes. In conclusion, the data indicate that serum TGF-β1 levels and Foxp3 (rs3761548) promoter polymorphism is not a risk factor for RSA and that there is no association between the polymorphism and serum TGF-β1 levels. [ABSTRACT FROM AUTHOR]

Background: Bone marrow mesenchymal stem cells (MSCs) are one of the undifferentiated multipotential cell sources of human body. MSCs have the capacity to form a variety of cell types, especially chondrocytes and osteocytes. Learning about responses of MSCs to external milieu and chemical factors such as pH could recommend new approaches for preparation of suitable scaffolds for bone and cartilage tissue engineering. In present study, the effect of alkaline medium on chondrogenic and osteogenic differentiation of rat MSCs was evaluated Methods: MSCs were harvested from bone marrow of animals and then the response of passage1 and 2 of MSCs (P1 MSCs & P2 MSCs) to the culture in alkaline medium (pH: 8) was evaluated. Cytochemical and immunocytochemical staining were performed to distinguish chondrocytes and osteocytes. Real-time PCR was performed to evaluate the type II collagen and osteopontin mRNA levels. Results: Staining for type II collagen, a chondrocytic specific marker, revealed that after one-week culture in alkaline medium, a considerable amount of P1 MSCs had shown chondrocytic morphology. By prolonging the culture period up to 4 weeks, osteogenic cells with expanded matrix and mineralized areas around them were appeared. Results of real-time PCR showed that P1 MSCs after one week culture in alkaline medium expressed highest rate of type II collagen and osteopontin mRNA among all groups. Conclusion: This study demonstrated that alkaline medium is a potent chondrogenic differentiation inducer for MSCs in their first passage. [ABSTRACT FROM AUTHOR]

Objective: Adipocyte and skeletal muscle are important tissues which contribute the development and progression of metabolic disorder. Insulin has a major regulatory function on glucose metabolism in these tissues by redistributing glucose transporter (GLUT4) from intracellular vesicles to the cell surface. Today, due to the side effects of chemical medications attendance to herbal medicines is growing. In this study, the effect of Curcumin extract as main polyphenols in Turmeric on gene expression of GLUT4 was evaluated. Materials and Methods: Curcumin was extracted using alcohol and chloroform from turmeric powder.TLC chromatography was used to confirm purity of Curcumin extracted. Mouse C2C12 myoblasts were grown in Dulbecco's Modified Eagle Medium (DMEM 1.5 g/l glucose) supplemented with 10% fetal bovine serum, 50 U/ml penicillin, and 50 μg/ml streptomycin. After differentiating of C2C12 cells during 4 days to myotubes, cells were treated separately in the presence of insulin (100 nM), Curcumin (25 μM) and co-treatment for 24h.RNA extraction from C2C12 cells was performed and GLUT4 expression levels were examined by semi-quantitative RT-PCR. Results: The results showed a significant increase in the GLUT4expression in Curcumin treatment group, in compare with negative control but less than insulin as a positive control. The synergistic effect of Curcumin and insulin was lower in comparison with insulin as a positive control. Conclusion: Curcumin could increase the gene expression but synergistic effects of Curcumin and insulin is more powerful than Insulin which can be due to the competitive action of insulin and Curcumin in activation of gene expression pathway. [ABSTRACT FROM AUTHOR]

By considering stem cell-based therapies as a new hope for the treatment of some tragic diseases, marrow stromal cells or marrow mesenchymal stem cells (MSCs) were considered as a suitable and safe multipotential cell source for this new therapeutic approach. For this purpose, many investigations have been performed on differentiation of MSCs toward specific cell lines to overcome the demand for providing the organ specific cells for cell therapy or preparation of engineered tissues. In the present study, differentiation of MSCs to endothelial cells (ECs) by mechanical and chemical stimulation was evaluated. Fluid shear stress (FSS) was used as mechanical inducer, while platelet lysate (PL) and estradiol (E) were used as chemical induction factors. MSCs were placed under FSS with different forces (2, 5 and 10 dyn/cm²) for different periods (6, 12 and 24 hours). In some groups, PL and E were added to the culture media to evaluate their effect on expression of EC specific markers. This investigation revealed that FSS with low tension (2.5-5 dyn/cm²) for a long time (24 hours) or high tension (10 dyn/cm²) in short time (6 hours) in the presence of PL could differentiate MSCs toward ECs. The presence of PL was necessary for initiation of endothelial differentiation, and in the absence of PL, there was not any expression of CD34 and Cadherin5 (Cdh5) among cells. Adding E to the culture medium did not change the rate of endothelial differentiation under FSS. Generated endothelial progenitors could produce von Willebrand factor (vWF) after two weeks culture and also they formed tubular structures after culture on matrigel. [ABSTRACT FROM AUTHOR]

Cutaneous leishmaniasis in Iran is usually caused by Leishmania major or L. tropica. However, the direct examination or the cultures of biopsies for diagnosis are not very sensitive. The objective of this study was to identify the responsible species obtained from patients suspected of cutaneous leishmaniasis referred to the reference laboratory at Yazd in Iran during 2010-2011 using parasitological and molecular assays. After completing a clinical/epidemiologic data questionnaire for 145 patients with suspected skin lesions, scraping samples were collected. Each specimen was examined using both direct microscopy and molecular assay using polymerase chain reaction-restriction length polymorphism (PCR-RFLP). Location of the lesions included 47.7% on hands, 30.7% on face, 15.4% on feet, and the remainder on other regions. Out of 145 samples, Leishman body was observed in 52 by direct smear and 73 by PCR assay. Molecular assay indicated 36 cases as L. major, 36 cases as L. tropica and one case as unknown. In conclusion, molecular characterization showed changing profile of Leishmania species in the study area which may have implications on treatment and/or control strategies. [ABSTRACT FROM AUTHOR]