Elucidating the impact of Y chromosome microdeletions and altered gene expression on male fertility in assisted reproduction.

BACKGROUND: Genetic abnormalities like Y chromosome microdeletions are implicated in male infertility. This study investigated the association of azoospermia factor (AZF) region microdeletions with unsuccessful assisted reproductive techniques (ART), including in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). METHODS: This cross-sectional analysis study examined 80 Iranian oligospermic men (mean age 34 years) with prior failed ICSI and IVF cycles (IR.IAU.TNB.REC.1401.041). Semen analysis evaluated quantity/quality parameters based on World Health Organization guidelines. Participants were stratified by sperm DNA fragmentation (SDF) levels into: control (SDF < 15%, n = 20), mild elevation (15% ≤ SDF ≤ 30%, n = 60), and high (SDF > 30%, n = 20). Multiplex PCR mapped AZF microdeletions in the high SDF group. The AZF-associated genes were selected by RNA Seq analysis, and the candidate genes were checked for expression level by real-time PCR. RESULTS: High SDF individuals exhibited poorer semen metrics, including 69% lower sperm concentration (P = 0.04) than those without SDF. Of this subset, 45% (9/20 men) harboured predominately AZF microdeletions. Men with AZF microdeletions showed higher SDF (32% vs 21%, P = 0.02) and altered AZF-associated genes expression. As USP9Y 3-fold, UTY 1.3-fold, and BPY2 1-fold revealed up-regulation, while IQCF1 8-fold, CDY 6.5-fold, DAZ 6-fold, and DDX3Y 1-fold underwent down-regulation. The PAWP gene was also down-regulated (5.7-fold, P = 0.029) in the IVF/ICSI failure group. CONCLUSION: AZF microdeletions significantly impact male infertility and ART outcomes. High SDF individuals exhibited poorer semen metrics, with 45% AZF microdeletions. These microdeletions altered AZF-associated genes expression, affecting fertility mediator PAWP independently. Dual AZF and SDF screening enables personalized management in severe male infertility, potentially explaining IVF/ICSI failures.

Suppression of B7-H7 Enhanced MCF-7 Cancer Cell Line's Chemosensitivity to Paclitaxel.

The B7-H7 is the newest addition to the B7 family of proteins that is present in the majority of malignancies. In this respect, the goal of the work was to investigate the impact of B7-H7 inhibition on breast cancer cells when paclitaxel and small interference RNA (siRNA) were combined. B7-H7 siRNA was used with Paclitaxel to treat MCF-7 cells. The IC50 of Paclitaxel and the cell survival was then assessed through using MTT assay. Investigation was conducted using flow cytometry to both the induction of apoptosis and the cell cycle. In addition, the clonogenic capacity of MCF-7 cells was investigated. Furthermore, qRT-PCR, was used to evaluate expression of genes. Our results demonstrated that suppressing B7-H7 sensitizes MCF-7 cells to Paclitaxel by triggering apoptosis and altering the expression of critical apoptosis mediator genes. In addition, the cell cycle was stopped in the sub-G1 and also G2-M phases as a result of the combination therapy leading prevention of developing colonies by MCF-7 cells. B7-H7 silencing improved the chemosensitivity of MCF-7 cells to Paclitaxel and demonstrated antiproliferative effects. After the adequate study has been conducted, this strategy may be regarded as a possible alternative treatment option for this cancer.

Association of Y chromosome AZF region microdeletions with recurrent miscarriage in Iranian couples: A case-control study.

BACKGROUND: Recent studies have linked recurrent pregnancy loss (RPL) to abnormalities in the sperm genome, specifically microdeletions in the azoospermia factor (AZF) region. This study investigated the potential association between Y chromosome microdeletions in the AZF region and RPL in Iranian couples. METHODS: The research presents a case-control study of 240 men: 120 whose partners experienced recurrent miscarriage, and 120 who had successful pregnancies without history of miscarriage. The study used semen parameters, hormone analyses, and microdeletion analysis via multiplex PCR and the YChromStrip kit. Thus, the sequence-tagged site (STS) markers of AZFa (sY84, sY86), AZFb (sY127, sY134), and AZFc (sY254, sY255) regions were examined. RESULTS: The variations in semen parameters and sex hormone levels between cases and controls are suggest impaired testicular function in men whose partners had recurrent miscarriages (p < 0.05). Furthermore, the study revealed a negative correlation between sperm count and follicle-stimulating hormone (FSH) level, and a positive one between sperm motility and testosterone concentration. There were no microdeletions in the control group, while the RPL group showed 20 deletions in AZFb (sY134) (16.66%) and 10 deletions each in AZFb (sY127) (8.33%) and AZFc (sY254) (8.33%). CONCLUSION: Microdeletions in sY134 (AZFb) were significantly associated with RPL in Iranian men (p = 0.03). AZF microdeletion screening in couples with RPL can provide valuable information for ethnical genetic counseling and management of recurrent miscarriage. Further studies on larger populations or across various ethnic groups, conclusions and the inclusion of other factors like epigenetic changes explain the role of AZF microdeletions in RPL.

Isolated Lactobacillus fermentum Ab.RS22 from traditional dairy products inhibits HeLa cervical cancer cell proliferation and modulates apoptosis by the PTEN-Akt pathway.

Objectives: It is worthwhile to note that, some probiotics such as Lactobacilli and Bifidobacteria isolated from dairy products have significant therapeutic effects against cancer cells. Here, we evaluated anti-proliferation and the apoptotic effects of isolated Lactobacillus fermentum Ab.RS22 from traditional dairy products on the HeLa cervical cancer cells in vitro. Materials and Methods: The viability of treated HeLa cells with supernatant of Lactobacillus in 0.5, 0.75, 1, 1.5, and 2 ng/ml concentrations, and IC50 values were detected by tetrazolium bromide. The L. fermentum Ab.RS22-induced cell death by flow cytometry was confirmed through evaluation of the expression of caspase-3, P53, PTEN, and AKT genes by quantitative reverse transcription-polymerase chain reactions (qRT-PCR). Results: Most cytotoxicity effects of Lactobacillus on HeLa cells were detected in 2 ng/ml at 24 hr (P<0.01); also, the IC50 value was measured as 1.5 ng/ml. The findings of the flow cytometry assay showed that L. fermentum Ab.RS22 in 1.5 ng/ml concentration at 24 hr increased the percentage of both apoptosis and necrosis cells. Lactobacillus-induced cell death was verified through results of Real-time PCR; where expression of caspase-3, P53, and PTEN genes was increased (P<0.01), and also expression of AKT gene (anti-apoptotic) was decreased (P<0.05). Conclusion: Our findings showed that L. fermentum Ab.RS22 could dose-dependently inhibit the proliferation of the HeLa cells. Its apoptotic effect was confirmed via modulating PTEN/p53/Akt gene expression and activation of the caspase-3 mediated apoptosis pathway. Therefore, L. fermentum Ab.RS22 can be considered a valuable anticancer candidate against cervical cancer progression in subsequent studies.

Tramadol induces apoptosis, inflammation, and oxidative stress in rat choroid plexus.

BACKGROUND: The choroid plexus (CP) is the principal source of cerebrospinal fluid (CSF). It can produce and release a wide range of materials, including growth and neurotrophic factors which have a crucial role in the maintenance and proper functioning of the brain. Tramadol is a synthetic analog of codeine, mainly prescribed to alleviate mild to moderate pains. Nevertheless, it causes several side effects, such as emotional instability and anxiety. METHODS: In this study, we focused on alterations in the expression of inflammatory and apoptotic genes in the CP under chronic tramadol exposure. Herein, rats were treated daily with tramadol at 50 mg/kg doses for three weeks. CSF samples were collected, with superoxide dismutase (SOD) and glutathione (GSH) measured in the CSF. RESULTS: We found that tramadol reduced the SOD and GSH levels in the CSF. Furthermore, the stereological analysis revealed a significant increase in the CP volume, epithelial cells, and capillary number upon tramadol administration. Tramadol elevated the number of blob mitochondria in CP. Also, we observed the upregulation of inflammatory and apoptosis genes following tramadol administration in the CP. CONCLUSIONS: Our findings indicate that tramadol induces neurotoxicity in the CP via apoptosis, inflammation, and oxidative stress.

The nano-micellar curcumin improves International Prostate Symptoms Score (IPSS) in patients with benign prostatic hyperplasia: a randomized clinical trial.

PURPOSE: Benign prostatic hyperplasia (BPH) is the main prevalent disorder in men over forty years, usually revealing itself with lower urinary tract symptoms. Despite the existence of different treatments, the incidence of BPH is increasing, so further studies for better management are a necessity. This research was designed to assay the effectiveness of nano-micellar curcumin on biomedical indicators of patients with BPH. METHODS: The present research was a double-blind, randomized, and placebo-controlled trial that enrolled fifty-two patients with BPH between June 2021 and December 2021. Participants were randomized to receive 160 mg/d nano-micellar curcumin (n = 26) or placebo (n = 26) as soft gel during 3 months. Primary end point was changes in International Prostate Symptoms Score (IPSS). Data gathering was occurred using a standard inquiry form and measuring other biomedical parameters based on routine laboratory techniques. To compare the distribution of demographics and covariates, independent t-test and Chi-square were used. RESULTS: Nano-micellar curcumin had significant effect on IPSS (p value: 0.010), low effect on high-sensitive C-reactive protein (hs-CRP) (p value: 0.032), and low to intermediate effect on malondialdehyde (MDA) (p value: 0.014) level as secondary end points after the intervention. The effect of nano-micellar curcumin on other parameters was negligible. CONCLUSION: Overall, this trial indicated 3-month intake of nano-micellar curcumin had considerable effects on IPSS as the most common clinical symptom and also two biomedical parameters including serum hs-CRP and MDA. TRIAL REGISTRATION: http://www.irct.ir : IRCT20170430033730N3.

Nanoimmunoengineering strategies in cancer diagnosis and therapy

Cancer immunotherapy strategies in combination with engineered nanosystems have yielded beneficial results in the treatment of cancer and their application is increasing day by day. The pivotal role of stimuli-responsive nanosystems and nanomedicine-based cancer immunotherapy, as a subsidiary discipline in the field of immunology, cannot be ignored. Today, rapid advances in nanomedicine are used as a platform for exploring new therapeutic applications and modern smart healthcare management strategies. The progress of nanomedicine in cancer treatment has confirmed the findings of immunotherapy in the medical research phase. This study concentrates on approaches connected to the efficacy of nanoimmunoengineering strategies for cancer immunotherapies and their applications. By assessing improved approaches, different aspects of the nanoimmunoengineering strategies for cancer therapies are discussed in this study (AU)

Nanoimmunoengineering strategies in cancer diagnosis and therapy.

Cancer immunotherapy strategies in combination with engineered nanosystems have yielded beneficial results in the treatment of cancer and their application is increasing day by day. The pivotal role of stimuli-responsive nanosystems and nanomedicine-based cancer immunotherapy, as a subsidiary discipline in the field of immunology, cannot be ignored. Today, rapid advances in nanomedicine are used as a platform for exploring new therapeutic applications and modern smart healthcare management strategies. The progress of nanomedicine in cancer treatment has confirmed the findings of immunotherapy in the medical research phase. This study concentrates on approaches connected to the efficacy of nanoimmunoengineering strategies for cancer immunotherapies and their applications. By assessing improved approaches, different aspects of the nanoimmunoengineering strategies for cancer therapies are discussed in this study.

The Remarkable Roles of the Receptor for Advanced Glycation End Products (RAGE) and Its Soluble Isoforms in COVID-19: The Importance of RAGE Pathway in the Lung Injuries.

The respiratory symptoms of acute respiratory distress syndrome (ARDS) in the coronavirus disease 2019 (COVID-19) patients is associated with accumulation of pre-inflammatory molecules such as advanced glycation end-products (AGES), calprotectin, high mobility group box family-1 (HMGB1), cytokines, angiotensin converting enzyme 2 (ACE2), and other molecules in the alveolar space of lungs and plasma. The receptor for advanced glycation end products (RAGEs), which is mediated by the mitogen-activated protein kinase (MAPK), plays a critical role in the severity of chronic inflammatory diseases such as diabetes mellitus (DM) and ARDS. The RAGE gene is most expressed in the alveolar epithelial cells (AECs) of the pulmonary system. Several clinical trials are now being conducted to determine the possible association between the levels of soluble isoforms of RAGE (sRAGE and esRAGE) and the severity of the disease in patients with ARDS and acute lung injury (ALI). In the current article, we reviewed the most recent studies on the RAGE/ligands axis and sRAGE/esRAGE levels in acute respiratory illness, with a focus on COVID-19-associated ARDS (CARDS) patients. According to the research conducted so far, sRAGE/esRAGE measurements in patients with CARDS can be used as a powerful chemical indicator among other biomarkers for assessment of early pulmonary involvement. Furthermore, inhibiting RAGE/MAPK and Angiotensin II receptor type 1 (ATR1) in CARDS patients can be a powerful strategy for diminishing cytokine storm and severe respiratory symptoms.

Short report of an Iranian Prenatal Methamphetamine Exposure effect cohort showed DNA methylation alteration of mitochondria function associated genes.

BACKGROUND: Use of Methamphetamine during pregnancy is significant public health concern since it affects the development of the brain and poor behavioral outcomes in children. Prenatal methamphetamine exposure (PME) may cause developmental disabilities and several gene expression and molecular pathways alterations. In the present study, DNA methylation of Propionyl-CoA Carboxylase subunit Beta (PCCB) and Protocadherin Alpha 12 (PCDHA12) genes were assessed in two groups of three-year-old children, those exposed to PME and healthy control children. AIMS: Clarification of PME role in methylation level of two mitochondria function associated genes; PCCB and PCDHA12. METHODS AND PROCEDURES: In this study, 2629 children with PME (1531male, 1098 female) and 3523(2077male, 1446 female) control children were recruited based on maternal self-report of prenatal exposure. Genomic DNA extracted from peripheral blood and pyrosequencing was used to determine the association between prenatal MA exposure and methylation in nine CpG sites of PCCB and PCDHA12 genes. OUTCOMES AND RESULTS: Prenatal methamphetamine exposure was associated with significant DNA hypomethylation of four out of five CpG sites in the PCCB gene and three out of four CpG sites in the PCDHA12 gene. Also, significant hypomethylation in the biding site of p53 transcription factor in PCCB gene was detected in children with PME. CONCLUSIONS AND IMPLICATIONS: Prenatal methamphetamine exposure is related to epigenetic alterations in PCCB and PCDHA12, as important mitochondria function associated genes. Detected hypomethylation in these genes was reported in neurodevelopmental and bioenergetics disabilities. It seems that PME could cause mitochondrial dysfunctions associated with developmental abnormalities. What this paper adds?

Activation of apoptosis and G0/G1 cell cycle arrest along with inhibition of melanogenesis by humic acid and fulvic acid: BAX/BCL-2 and Tyr genes expression and evaluation of nanomechanical properties in A375 human melanoma cell line.

Objectives: Humic acid (HA) and Fulvic acid (FA) are major members of humic substances, which are extracted from organic sources including soil and peat. The pro-apoptotic and anti-melanogenic effects of HA and FA at the cellular and molecular levels in the A375 human melanoma cell line were examined in this study. Materials and Methods: The cytotoxicity effect of HA and FA were evaluated by cell viability assay. Apoptosis and cell cycle were investigated by flow cytometry. Real-time PCR was carried out to measure the expression of BAX, BCL-2, and Tyr genes. Moreover, the changes in nanomechanical properties were determined through atomic force microscopy (AFM). Results: It was found that HA and FA decrease cell viability with an IC50 value of 50 µg/ml (dose-dependent) for 14 hr, arrested cells in the G0/G1 phase, and increased the sub-G1 phase (induce apoptosis). Based on the AFM analysis, Young's modulus and adhesion force values were increased, also ultrastructural characteristics of cells were changed. Results of Real-time PCR revealed that HA and FA lead to a decrease in the expressions of BCL-2 and Tyr genes, and increase the BAX gene expression. Conclusion: These results exhibited that HA and FA possess pro-apoptotic effects through increasing the BAX/ BCL-2 expression in A375 cells. These molecular reports were confirmed by cellular nanomechanical assessments using AFM and flow cytometry. In addition, HA and FA inhibited melanogenesis by decreasing the expression of the Tyr gene. It is worthwhile to note that, HA and FA can be regarded to design new anti-cancer and anti-melanogenesis products.

Prevalence and molecular assessment of Sarcocystis infection in livestock in northeast Iran.

Sarcocystis is an intracellular parasite of the apicomplexa phylum. More than one hundred species of Sarcocystis can infect wildlife and livestock animals. Samples of the liver, heart, muscle and diaphragm were collected from sheep, goats and cattle in Gonabad, northeast Iran and subjected to macroscopic, microscopic, tissue digestion, sequencing and phylogenetic analyses of 18S-rRNA region. Tissue sections stained with hematoxylin and eosin were also provided for surveying sarcocysts in the samples. Tissue digestion showed that all animal samples (100%) were infected with Sarcocystis bradyzoites. Phylogenetic analysis indicated that out of 50% of sheep genotypes belonged to S. tenella, and 20% to S. moulei and S. arieticanis. Moreover, three samples of macroscopic specimens of sheep were identified as S. gigantea, 100% of cattle isolates infected with S. cruzi, 80% of goat isolates belonged to S. capracanis and one macroscopic specimen of goat (20%) identified as S. moulei. Sarcocystis infection is highly prevalent in livestock in northeast Iran, where a variety of Sarcocystis species are unequivocally circulating in the region.

Prevalence and Subtype Analysis of Blastocystis hominis Isolated from Patients in the Northeast of Iran.

Blastocystis hominis is the most common intestinal parasite found in humans and many other hosts. Pathogenicity of Blastocystis spp. remains controversial, and it has been suggested that it may be associated with specific subtypes of the organism. This study identified the B. hominis subtypes and their prevalence rates in the northeast of Iran. A total of 1878 samples were collected from the northeast of Iran from January to December 2017. The patients' demographic details were recorded. Samples were examined by a wet mount, and genomic DNA was extracted from positive samples. Also, PCR was done on the positive samples, and sequencing and phylogenetic analysis were subsequently performed. From 1878 collected stool samples, 152 (8.1%) Blastocystis samples were detected by the microscopic method. Of the 152 samples, Blastocystis spp. were found in 53.6% of the men and 28.9% of the women who showed clinical gastrointestinal symptoms, and a significant relationship was observed between gender and clinical symptoms (P = 0.002). A meaningful relationship was found between the season and infection with this parasite (P value = 0.003). The results of the sequencing of 22 PCR products showed the dominance of ST3, which was isolated from 10 (45.45%) patients, while ST1, ST2, and ST7 were found in 4 (18.19%), 7 (31.81%), and 1 (4.55%) patients, respectively. In this study, ST7 had a low prevalence in the northeast of Iran, and similar to previous studies, ST3 was the dominant subtype.

Effects of Coenzyme Q10 and Diamond Nanoparticles on Ischemia-Reperfusion-Induced Testicular Damages in Rats.

Background: Ischemia-reperfusion (I/R) induced by testicular torsion can damage the testicles. In the present study, we assessed the effects of coenzyme Q10 (CoQ10) and diamond nanoparticles on sperm parameters in I/R testes in rats. Materials and Methods: Forty-eight Wistar adult male rats were divided into eight groups: healthy control (Ch), diamond nanoparticle healthy control group (Ch+Dia), CoQ10 healthy control group (Ch+Q10), diamond nanoparticles+CoQ10 healthy control group (Ch+Q10+Dia), torsion/detorsion group (Ct), the Ct group that received diamond nanoparticles (Ct+Dia), the Ct group that received CoQ10 (Ct+Q10), and Ct group that received diamond nanoparticles and CoQ10 (Ct+Q10+Dia). The rats were euthanized, and we collected the semen from the epididymal tissues to evaluate sperm viability, motility, concentration, and morphology parameters. Results: The I/R of the testicles significantly reduced sperm concentration, motility, viability, and altered sperm morphology in the rats. However, the administration of CoQ10 significantly improved sperm parameters in the rats with testicular I/R. Diamond nanoparticles decreased the sperm parameters; however, simultaneous administration of diamond nanoparticles and CoQ10 led to improved sperm parameters. Conclusion: CoQ10 potentially appeared to have protective effects against the long-term side-effects of I/R in testes in rats. Co-administration of diamond nanoparticles with CoQ10 significantly improved sperm parameters and greatly reduced the negative effects of diamond nanoparticles alone. Therefore, green synthesis of nanoparticles with the use of antioxidants such as CoQ10 is recommended.

Investigation of Toxoplasma gondii Infection in Aborted Fetuses of Sheep Using PCR: A Study in North Khorasan Province, Iran.

Toxoplasma gondii is a zoonotic obligate intracellular protozoan parasite that infects warm-blooded animals as well as humans worldwide. The purpose of this study was to delineate the prevalence of Toxoplasma infection in aborted fetuses of sheep in North Khorasan province, Iran. Three hundred and ninety-nine samples of the liver (133 samples), placenta (133 samples), and brain (133 samples) from 133 aborted fetuses of sheep were collected from 2015 to 2017. The ages of aborted fetuses were higher than 120 days' gestational age in this study. According to the samples, sixteen out of 133 aborted fetuses of sheep were infected with T. gondii. Toxoplasma DNA was found in the placenta (68.75%) and liver (31.25%) samples of infected fetuses using the PCR method. The highest and lowest rates of Toxoplasma infection were observed during 2016 and 2017, respectively. Shirvan and Faruj provinces were recognized as the two most infected districts among others. There was a significant difference between the year and abortion rate in sheep due to infection by the Toxoplasma parasite (P < 0.05). Furthermore, no significant difference between the prevalence of T. gondii infection and aborted fetuses was seen (P > 0.05) in different areas. According to the present study, T. gondii infection can be one of the causes of fetus abortion of sheep in North Khorasan province, Iran.

Detection and characterization of purified antigenic proteins from culture filtrate of Mycobacterium bovis strain AN5.

BACKGROUND AND OBJECTIVES: Bovine tuberculosis diagnosis is usually performed by various tests with specific limitations. Mycobacterium bovis culture filtrate contains antigenic proteins that could be used to improve the sensitivity of bovine tuberculosis diagnosis. The objective of this study was to identify and purify antigenic proteins from culture filtrates of M. bovis strain AN5 for use in immunological assays. MATERIALS AND METHODS: Secreted proteins were purified from the heat-treated culture filtrate of M. bovis strain AN5. Proteins were precipitated with ammonium sulfate, fractionated by Sephadex G50 chromatography. The protein concentrations and the approximate molecular weight were determined by lowry method and 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), respectively. Immunological methods, including dot-blotting and western blotting, assessed the quality of the isolated proteins. RESULTS: The quantity of antigenic proteins in the culture medium was measured at far more than 15% of the amount of proteins secreted into medium. Three main chromatographic fractions obtained and showed concentrations of proteins ranging from 14 to 60 µg/ µl with molecular weights in the 10 to 180 kDa range. The purified antigens showed positive reactions to the infected cattle serum throughout dot-blotting. Western blotting revealed a total of 15 to 70 kDa molecular weight proteins. CONCLUSION: Immunoblotting analysis made it possible to detect and recognize novel antigens that are useful for bovine tuberculosis diagnosis improvement. This is significant since non-specific reactions were not observed when we utilized serum of cattle experimentally infected with M. bovis as a polyclonal antibody.

SNP Scanning in mecA Gene for Methicillin-Resistant Staphylococcus aureus.

BACKGROUND: Staphylococcus aureus (SA) is known as an important human pathogen, which is responsible for many cases of both hospital and community-acquired infections all over the world. Studying on drug resistance is regarded as an important prevention strategy regarding these types of infections. OBJECTIVES: The current study is aimed to assess the association between the single-nucleotide polymorphism (SNP) and resistance to antibiotics in the methicillin-resistant Staphylococcus aureus (MRSA) strains as well as the molecular typing of isolates, collected from the clinical samples. MATERIALS AND METHODS: We used the disc-diffusion method to test the isolates antibiotic resistance. In addition, the genotypes of staphylococcal cassette chromosome mec (SCCmec) in the Methicillin-resistant Staphylococcus aureus isolates were determined by multiplex -polymerase chain reaction (PCR). SNP was identified in the mecA gene using sequencing and amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) method. RESULTS: The highest resistance was shown against oxacillin, and erythromycin and cephalexin. The most sensitive antibiotic was vancomycin (97%) and resistance to at least three antibiotic classes were identified in all isolates. Eighty six percent of isolates were positive for mecA gene and more than 50% of which were healthcare-acquired methicillin-resistant Staphylococcus aureus (HA-MRSA). Moreover, SCCmec type 3, 1were the predominant strains of the identified MRSA. Also, 23 isolates (23%) were non-typable. By using the ARMS-PCR method, it was found that 10% of the clinical specimens had SNP in the mecA gene. CONCLUSION: According to the Chi-square test (χ2), it reveals that the association between SNP in the mecA gene and oxacillin, cefoxitin, and erythromycin resistance was confirmed among clinical MRSA. Furthermore, there is a 95%probability of association between SNP and resistance to more than three antibiotics in MRSA strains.

Isolation and genotyping of Acanthamoeba strains from water sources of Kermanshah, Iran

Acanthamoeba spp. are free-living amoeba commonly found in environmental sources such as soil, water, and dust. This ubiquitous amoeba is the causative agent of amoebic keratitis (AK) and encephalitis. The present study aimed to investigate the presence of Acanthamoeba spp. in the water sources of Kermanshah city, Iran. Sixty water samples were taken from different localities of Kermanshah including agricultural canals, rivers, and swimming pools. Filtration and cultivation were carried out on non-nutrient agar medium (NNA). The axenic cultivation was performed for all of the positive isolates. PCR analysis was performed on positive samples. Sequencing was done for 12 PCR products. Genotypes were identified by blast search and homology analysis. The obtained data were analyzed using Statistical Package for the Social Sciences (SPSS 16) software. Acanthamoeba spp. was found in 46 (76.66%) water samples and amoebae were grown in the TYI-S-33 medium. Sequencing of 12 samples proved that Acanthamoeba belonged to T4 (75%), T2 (8.34%), T5 (8.33%) and T11 (8.33%) genotypes. In this study, Acanthamoeba T4 (75%), T2 (8.34%), T5 (8.33%) and T11 (8.33%) genotypes were isolated from the water of Kermanshah city. Thus, hygiene consideration is recommended to prevent the contamination.