Evaluation of antibacterial activity of five biocides and the synergistic effect of biocide/EDTA combinations on biofilm-producing and non-producing Stenotrophomonas maltophilia strains isolated from clinical specimens in Iran.

BACKGROUND: The overuse of biocides in healthcare-facilities poses risk for emergence and spread of antibiotic resistance among nosocomial pathogens. Hospital-acquired infections due to S. maltophilia have been increased in the recent years and with its various resistance mechanisms contribute to patient morbidity and mortality in hospitals. The current study aimed to evaluate the susceptibility of biofilm-producing and non-producing S. maltophilia clinical isolates to five commonly used hospital biocides, alone and in combination with EDTA to examine the synergistic effect of combining EDTA on the bactericidal activity of them by microbroth dilution method. As well as the frequency of efflux genes encoding resistance to biocides among isolates. This study also intended to assess the effect of exposure of S. maltophilia isolates to sub-inhibitory concentrations of sodium hypochlorite upon the antimicrobial susceptibility patterns. RESULTS: Based on minimum inhibitory and bactericidal concentrations of biocides sodium hypochlorite 5% (w/v) and ethyl alcohol 70% (v/v) were the strongest and weakest biocides against S. maltophilia isolates, respectively. The combination of EDTA with biocides significantly increased the effectiveness of the studied biocides. Exposure to sub-inhibitory concentration of sodium hypochlorite showed a significant change in the susceptibility of isolates towards ceftazidime (p = 0.019), ticarcillin/clavulanate (p = 0.009), and chloramphenicol (p = 0.028). As well as among the isolates examined, 94 (95%) were able to produce biofilm. The frequency of sugE1 resistance genes was found in 90.7% of our clinical S. maltophilia isolates. None of the isolates carried qacE and qacEΔ1 gene. CONCLUSIONS: The current study recommended that using the mixture of biocides with EDTA can be effective in reducing nosocomial infections. Also, this study demonstrated that exposure to sub-inhibitory concentrations of sodium hypochlorite leads to reduced antibiotic susceptibility and development of multidrug-resistant S. maltophilia strains.

Detection of bacterial agents causing prostate infection by culture and molecular methods from biopsy specimens.

Background and Objectives: Prostatitis affects about 16% of men in their lifetime and sometimes leading to prostate cancer. Bacterial infections are the most common causes of prostatitis. Diagnosis of the causative agents of bacterial prostate infections plays an essential role in timely treating and preventing secondary complications. This study isolated bacterial infectious agents in patients' surgical prostate and evaluated them by routine and molecular microbiological methods. Materials and Methods: In this cross-sectional study, 72 prostate biopsy specimens were collected from the Orology Departmen of hospitals of Qazvin University of Medical Sciences. All samples were cultured in aerobic and anaerobic conditions. Antibiotic susceptibility test by Kirby-Bauer standard method was performed for all isolated bacteria. In addition, all isolated bacteria were identified using 16S rDNA PCR and sanger sequencing methods. Also, TaqMan real-time PCR was applied to detect Ureaplasm aurealyticum, Mycoplasma hominins, and Mycoplasma genitalium. Results: In conventional culture method, out of 18 positive samples, 15 samples (83.3%) were Gram-negative bacteria and 3 samples (16.6%) were Gram-positive bacteria, containing Escherichia coli (55.5%), Klebsiella pneumoniae (11.1%), Enterobacter cloacae (5.5%), Pseudomonas aeruginosa (11.1%), Staphylococcus aureus (11.1%), and Enterococcus faecalis (5.5%). The results of molecular identification methods were the same as conventional culture results. Also, four patients were Ureaplasm aurealyticum, and three patients were positive for Mycoplasma hominis. Conclusion: Most bacteria isolated from prostate specimens belonged to the Enterobacteriaceae family, especially Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae. Staphylococcus aureus and Enterococcus faecalis were cocci isolated in the specimens too. Also, Ureaplasma urealyticum, and Mycoplasma hominis were identified in prostatitis.

Phenotype and genetic determination of resistance to common disinfectants among biofilm-producing and non-producing Pseudomonas aeruginosa strains from clinical specimens in Iran.

BACKGROUND: Pseudomonas aeruginosa is a common pathogen in Hospitalized patients, and its various resistance mechanisms contribute to patient morbidity and mortality. The main aims of the present study were to assess the susceptibility of biofilm-producing and non-producing P. aeruginosa isolates to the five commonly used Hospital disinfectants, to evaluate the synergistic effect of selected disinfectants and Ethylene-diamine-tetra acetic acid (EDTA), and the effect of exposure to sub-inhibitory concentrations of Sodium hypochlorite on antimicrobial susceptibility test. RESULTS: The results showed that sodium hypochlorite 5% and Ethanol 70% were the most and least effective disinfectants against P. aeruginosa, respectively. The addition of EDTA significantly increased the effectiveness of the selected disinfectants. The changes in the antibiotic-resistance profiles after exposure to sub-inhibitory concentrations of disinfectants were observed for different classes of antibiotics (Carbapenems, Aminoglycosides, Cephalosporins, Fluoroquinolones). As well as near the all isolates harbored efflux pump genes and 117 (97.5%) of isolates produced biofilm. CONCLUSION: In the current study, the mixture of disinfectant and EDTA were the most suitable selection to disinfect Hospital surfaces and instruments. Also, it was clear that exposure to sub-inhibitory concentrations of Sodium hypochlorite results in resistance to some antibiotics in P. aeruginosa species. Strong and intermediate biofilm formers belonged to MDR/XDR strains. Future studies should include more complex microbial communities residing in the Hospitals, and more disinfectants use in Hospitals.

Screening and molecular characterization of Trichomonas vaginalis genotypes isolated from married women in northern Iran.

Trichomonas vaginalis is an anaerobic protozoan parasite that causes trichomonosis in human. It is one of the most common non-viral sexually transmitted infections. It has been found to be most prevalent in patients referred to sexually transmitted disease clinics. In recent years, molecular methods have been used to identify genotypes of this parasite in different parts of the world and so far 6 types of T. vaginalis have identified. The aim of this study was to investigate the prevalence and genotype identification of T. vaginalis from married women in northern Iran. A total of 450 vaginal specimens were taken from married women, referring to health centers in northern Iran. Demographic information of women was collected through a questionnaire. The samples were first examined microscopically and then monitored in Dorsch culture medium for up to 10 days. Actin genes of positive samples were amplified by PCR. Finally, PCR products were used to determine the sequence and genotype of the parasite. Overall, 0.7% (3/450) samples were positive for T. vaginalis. All of the three infected women were housewives. After sequencing, the genotype of these parasites were type H (66.7%) (Accession no; MW414672-MW414673) and type E (33.3%) (Accession no: MW414671). Low prevalence of T. vaginalis in north of Iran indicate high level of hygiene in sexual intercourse and avoiding from high risk sexual behaviors, and also it seems that genotype H is dominant type of the parasite in the study area.

Development of a SYBR Green Multiplex Real Time PCR for Simultaneous Detection of Mycobacterium Tuberculosis and Nocardia Asteroides in Respiratory Samples.

BACKGROUND: Nocardia asteroides and Mycobacterium tuberculosis are worldwide-distributed bacteria. These infectious agents can cause many infections in humans, especially in immunocompromised individuals. Pulmonary infections are more common and have similar clinical symptoms. Proper diagnosis and treatment of these patients are important for accurate treatment and could be lifesaving. METHODS: In this study, a multiplex real-time PCR assay was established for the simultaneous detection of the N. asteroides and M. tuberculosis. Both this homemade multiplex real time PCR and routine commercial tuberculosis tests were performed on 150 pulmonary specimens collected from individuals suspected to have tuberculosis. RESULTS: From 150 specimens, 20 samples were acid fast positive, 14 positives for M. tuberculosis by singleplex real time PCR, 10 positives for N. asteroides by singleplex real time PCR and 2 positives for M. tuberculosis and N. asteroides by multiplex real time PCR whereas 14 samples were positive for M. tuberculosis with commercial test. Differential diagnosis of pulmonary tuberculosis is useful for their proper treatment. CONCLUSION: Our test had good performance for differential diagnosis of tuberculosis and nocardiosis. Therefore, it is recommended to be used to diagnose such patients.

Isolation and Molecular Identification of Acanthamoeba and Naegleria from Agricultural Water Canal in Qazvin, Iran.

BACKGROUND: Free-living amoeba (FLA) are widely distributed in different environmental sources. The most genera of the amoeba are Acanthamoeba, Naegleria and Vermamoeba. The most common consequences of the infections in immune-deficient and immuno-competent persons are amoebic encephalitis and keratitis. The aim of this study was to investigate the presence of Acanthamoeba spp. and Naegleria spp., isolated from the main agricultural water canal in Qazvin. METHODS: Totally, 120 water specimens were collected and later the specimens were cultured and cloned to identify positive samples. PCR amplification and sequencing were carried out to identify the isolated species as well as the genotypes of amoeba. RESULTS: According to morphological surveys, 41.7% (50/120) of water specimens were positive for FLA. Molecular analysis revealed that 68.6% and 31.4% of Acanthamoeba specimens were identified as T3 and T4 genotypes, respectively. Also, two species of Naegleria named as N. lovaniensis (57.1%) and Naegleria sp. (42.8%) were identified. The results of pathogenicity assays demonstrated that 38.5% of T3 and 61.5% of T4 genotypes of Acanthamoeba were highly pathogenic parasites. CONCLUSION: The water flowing in the agricultural canal of the area is contaminated with potential pathogenic FLA, therefore, it is recommended that more attention to be paid towards proper treatment of water sources to prevent possible risk of the disease.

Multifunctional betanin nanoliposomes-incorporated gelatin/chitosan nanofiber/ZnO nanoparticles nanocomposite film for fresh beef preservation.

The objective of this study was to fabricate betanin nanoliposomes incorporated gelatin/chitosan nanofiber/ZnO nanoparticles bionanocomposite film (G/CH NF/ZnO NPs/B NLPs) and investigate its effects on the preservation of fresh beef. The scanning electron microscopy image of nanocomposite film displayed a good inter-connective porous morphology. Fourier transform infrared and X-ray diffraction analysis confirmed the formation of new hydrogen bonds and enhanced crystallinity through the addition of CH NF, ZnO NPs, and B NLPs. The G/CH NF/ZnO NPs/B NLPs film exhibited satisfactory mechanical properties and high surface hydrophobicity (water contact angle = 92.49 ± 3.71°). The incorporation of ZnO NPs and B NLPs in the nanocomposite film provided high antibacterial activity and DPPH inhibition activity (53.02 ± 3.26%). The growth of inoculated bacteria, lipid oxidation, and the changes in the pH and color quality of the beef samples were controlled by packaging with the fabricated film. In conclusion, the G/CH NF/ZnO NPs/B NLPs nanocomposite has a high potential for meat preservation.

Molecular epidemiology of Blastocystis spp. in children referred to Qods hospital in northwest of Iran.

Blastocystis sp. is a polymorphic intestinal parasite in humans and animals. The parasite has a worldwide distribution, especially in developing countries with poor sanitation, exposure to animals, and improper disposal systems. The aim of this study was to identify the subtypes of Blastocystis sp. among children of Qazvin, northwest Iran. Totally, 864 stool samples were collected from the children referred to Qods hospital in Qazvin, Iran. Fecal specimens were investigated by formalin-ethyl acetate concentration method and trichrome staining as well as cultivation of all samples in clotted fetal bovine medium. DNA extraction of culture-positive specimens and PCR amplification of 18S ribosomal RNA gene region was performed. The sequences detected were compared with reference genes in the GenBank, and the sequences further deposited in the GenBank database. Data analysis was performed by Chi square test while a p value of < 0.05 was considered as significant. Of 864 isolates, 4.1% (36/864) were positive for Blastocystis sp. with infection rate insignificantly higher among the females than males. The highest infection rate was estimated at 6.8% in 6-9 years old age group with abdominal pain as the most common (33%) gastrointestinal sign. No statistically significant difference was found between the variables and Blastocystis infection. Molecular analysis clarified the presence of three subtypes of Blastocystis including ST1 (56%), ST2 (28%), and ST3 (16%) of among specimens with ST1 as the predominant subtype. A significant association between intestinal signs and the subtypes was not found. Considering ST1 as the predominant subtype, it seems that zoonotic transmission is a main route of human infections with Blastocystis sp. in the area studied.

Distribution and molecular analysis of Blastocystis subtypes from gastrointestinal symptomatic and asymptomatic patients in Iran.

INTRODUCTION: Blastocystis is a common intestinal parasite of human and animal hosts. The parasite has 17 subtypes, and among those at least nine subtypes (ST1-ST9) are found in human hosts. OBJECTIVE: The aim of the present study was to investigate the presence of different subtypes of Blastocystis spp. among the patients referred to Velayat hospital of Qazvin province, Iran. METHODS: Overall, 864 stool samples were examined by using formalin-ethyl acetate concentration method and Trichrome staining. All specimens were cultured in clotted fetal bovine medium. Later, DNA extraction and PCR amplification of 18S ribosomal RNA gene region was conducted and phylogenetic tree constructed. RESULTS: The results revealed 7.9% (68/864) of the study population were infected with Blastocystis. Intestinal symptoms were observed in 61% (36/59) of individuals positive for Blastocystis, with abdominal pain in 58% (21/36) of cases which was more frequent than other intestinal signs. No significant relationship was observed among the study variables. By molecular and phylogenetic analysis, three subtypes ST1 (45%), ST2 (30%) and ST3 (23%) of parasite were identified. CONCLUSION: This study showed ST1 subtype was the predominant subtype among the positive specimens, meanwhile the highest haplotype and nucleotide diversity were clarified in ST3 subtype.

Molecular Characterization of Fasciola and Dicrocoelium Species Isolated from Ruminant Livestock in Qazvin, Iran.

INTRODUCTION: Fascioliasis and dicrocoeliasis are the most frequent zoonotic diseases with increasing human health problems in different parts of Iran. Two species, Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica), are spread in the country. Molecular approaches have a decisive role in identifying both the species. The aim of this study was to detect Fasciola spp. and Dicrocoelium spp. by amplifying the ITS-2 and 28S rDNA gene sequence. METHODS: Overall, 30 infected liver samples were collected from the livestock of Qazvin, Iran. The adult flukes were collected from different livestock. DNA extraction and PCR amplification of ribosomal RNA gene region (ITS2) and 28S rDNA gene fragment were conducted and a phylogenetic tree was constructed. RESULT: All the isolates obtained from the cattle (No: 7) and 82.6% (No: 19) of sheep isolates were infected with F. hepatica species, whereas 17.4% (No: 4) of sheep isolates were infected with F. gigantica. It was also shown that F. hepatica was the predominant species of Fasciola present in the region. All the specimens were infected with Dicrocoelium dendriticum (D. dendriticum). CONCLUSION: Both the species of Fasciola were found in Qazvin. D. dendriticum was the sole infecting species of the Dicrocoelium genus in the livestock of the city of Qazvin. Further research studies are needed to determine the intermediate host of the parasites in the region.

Probiotic mixture of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 attenuates hippocampal apoptosis induced by lipopolysaccharide in rats

In recent years, the beneficial impact of targeted gut microbiota manipulation in various neurological disorders has become more evident. Therefore, probiotics have been considered as a promising approach to modulate brain gene expression and neuronal pathways even in some neurodegenerative diseases. The purpose of this study was to determine the effect of probiotic biotherapy with combination of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 on the expression levels of proteins critical to neuronal apoptosis in hippocampus of lipopolysaccharide (LPS)-exposed rats. Four groups of animals (Control, LPS, Probiotic + LPS, and Probiotic) were treated with maltodextrin (placebo) or probiotic (109 CFU/ml/rat) for 2 weeks by gavage. On the 15th day, a single intraperitoneal dose of saline or LPS (1 mg/kg) was injected and 4 h later, protein assessment was performed by western blotting in hippocampal tissues. LPS significantly increased the Bax, Bax/Bcl-2 ratio, and cleaved caspase-3 expression along with decreased the Bcl-2 and procaspase-3 protein levels. However, probiotic pretreatment (L. helveticus R0052 + B. longum R0175) significantly downregulated the Bax and Bax/Bcl-2 ratio accompanied with upregulated Bcl-2 expression. Prophylactic treatment with these bacteria also attenuated LPS-induced caspase-3 activation by remarkably increasing the expression of procaspase-3 while reducing the level of cleaved caspase-3 in target tissues. Our data indicate that probiotic formulation (L. helveticus R0052 + B. longum R0175) alleviated hippocampal apoptosis induced by LPS in rats via the gut-brain axis and suggest that this probiotic could play a beneficial role in some neurodegenerative conditions

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Molecular characterization of Hymenolepis nana based on nuclear rDNA ITS2 gene marker.

INTRODUCTION: Hymenolepis nana is a zoonotic tapeworm with widespread distribution. The goal of the present study was to identify the parasite in the specimens collected from NorthWestern regions of Iran using PCR-sequencing method. METHODS: A total of 1521 stool samples were collected from the study individuals. Initially, the identification of hymenolepis nana was confirmed by parasitological method including direct wet-mount and formalin-ethyl acetate concentration methods. Afterward, PCR-sequencing analysis of ribosomal ITS2 fragment was targeted to investigate the molecular identification of the parasite. RESULTS: Overall, 0.65% (10/1521) of the isolates were contaminated with H. nana in formalin-ethyl acetate concentration. All ten isolates were succefully amplified by PCR and further sequenced. The determined sequences were deposited in GenBank under the accession numbers MH337810 -MH337819. CONCLUSION: Our results clarified the presence of H. nana among the patients in the study areas. In addition, the molecular technique could be accessible when the human eggs are the only sources available to identify and diagnose the parasite.

Probiotic mixture of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 attenuates hippocampal apoptosis induced by lipopolysaccharide in rats.

In recent years, the beneficial impact of targeted gut microbiota manipulation in various neurological disorders has become more evident. Therefore, probiotics have been considered as a promising approach to modulate brain gene expression and neuronal pathways even in some neurodegenerative diseases. The purpose of this study was to determine the effect of probiotic biotherapy with combination of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 on the expression levels of proteins critical to neuronal apoptosis in hippocampus of lipopolysaccharide (LPS)-exposed rats. Four groups of animals (Control, LPS, Probiotic + LPS, and Probiotic) were treated with maltodextrin (placebo) or probiotic (109 CFU/ml/rat) for 2 weeks by gavage. On the 15th day, a single intraperitoneal dose of saline or LPS (1 mg/kg) was injected and 4 h later, protein assessment was performed by western blotting in hippocampal tissues. LPS significantly increased the Bax, Bax/Bcl-2 ratio, and cleaved caspase-3 expression along with decreased the Bcl-2 and procaspase-3 protein levels. However, probiotic pretreatment (L. helveticus R0052 + B. longum R0175) significantly downregulated the Bax and Bax/Bcl-2 ratio accompanied with upregulated Bcl-2 expression. Prophylactic treatment with these bacteria also attenuated LPS-induced caspase-3 activation by remarkably increasing the expression of procaspase-3 while reducing the level of cleaved caspase-3 in target tissues. Our data indicate that probiotic formulation (L. helveticus R0052 + B. longum R0175) alleviated hippocampal apoptosis induced by LPS in rats via the gut-brain axis and suggest that this probiotic could play a beneficial role in some neurodegenerative conditions.

A serologic study on Toxoplasma gondii infection in slaughtered sheep and goats in Qazvin Province, Iran.

BACKGROUND: Toxoplasma gondii is a common protozoan parasite among all mammals, in particular small ruminants, worldwide. Traditional husbandry can be a major risk factor for infection of sheep and goats with this parasite. OBJECTIVES: The present study aimed to determine the current status of the prevalence for T. gondii in livestock of Qazvin Province. METHODS: In this cross-sectional study, the sera of 455 sheep and 375 goats were examined to detect anti-Toxoplasma IgG antibodies by using in-house indirect ELISA. RESULTS: Overall, 33.62% (153/455) of sheep and 36.41% (130/375) of goats were positive for anti-Toxoplasma IgG antibodies with no statistically significant difference. The prevalence rate of T. gondii among the sheep of Qazvin County was significantly higher than in Abyek and Abhar counties (p < 0.001). CONCLUSIONS: The results of the present study indicate that the prevalence of T. gondii in sheep and goats of the study area is high. Therefore, the meat of the animals reared in this area can be a potential source of human infections by this parasite.

Investigation of Giardia intestinalis Genotypes among the Food Handlers of Qazvin, Iran.

BACKGROUND: We aimed to investigate the genotypes of Giardia intestinalis among the food handlers in Qazvin, Iran. METHODS: Overall, 1530 stool specimens were collected from the food handlers who visited Shahid Bolandian Health Center, Qazvin, Iran during 2016. Specimens were evaluated by microscopic and concentration methods. Twenty specimens with appropriate number of giardia cysts were selected followed by DNA extraction. Determination of giardia genotypes was achieved through PCR and sequencing the glutamate dehydrogenase gene. The phylogenetic tree was drawn using the MEGA7 software. Finally, the data were analyzed statistically with a P-value<0.05 was considered as significant. RESULTS: Twenty stool samples (1.3%) were positive for Giardia cyst. All positive specimens were obtained from male participants with abdominal cramp being their most common symptoms. The mean age for infected individuals was 32 yr. Molecular characterization was successfully performed for 17 isolates and two genotypes A (AII, 65%) and B (BIII, 35%) were identified. CONCLUSION: The most prevalent giardia genotypes among the food handlers in Qazvin were A (AII) and B (BIII) genotypes with A (AII) genotype as the dominant one in the region. Considering the direct association between the food handlers and public health as well as the impact of geographical and host conditions on dispersion and pathogenicity of various genotypes and their zoonotic aspects, further investigations are necessary.

The Effects of Probiotic Formulation Pretreatment (Lactobacillus helveticus R0052 and Bifidobacterium longum R0175) on a Lipopolysaccharide Rat Model.

OBJECTIVE: The role of gut microbiota in the pathogenesis of several neurodegenerative disorders, including Alzheimer's disease (AD), via the gut-brain axis has recently been demonstrated; hence, modification of the intestinal microbiota composition by probiotic biotherapy could be a therapeutic target for these conditions. The aim of this study was to assess the effects of a probiotic formulation (Lactobacillus helveticus R0052 and Bifidobacterium longum R0175) on inflammatory and memory processes in lipopolysaccharide (LPS)-induced rats, one of the animal models used in peripherally induced neuroinflammation and neurodegeneration. METHODS: Rats were randomly divided into four groups (Control, LPS, Probiotic + LPS, and Probiotic). All experimental groups were orally administrated maltodextrin (placebo) or probiotic (109 CFU/ml/rat) for 14 consecutive days and then were injected with saline or LPS (1 mg/kg, intraperitoneally [i.p.], single dose) 20 hours later. Memory retention ability and systemic and neuroinflammatory markers were assessed 4 hours after the injections. RESULTS: Systemic exposure to LPS resulted in significant elevation of both the circulating and hippocampal levels of proinflammatory cytokines, which decreased remarkably following probiotic pretreatment. Oral bacteriotherapy with a combination of L. helveticus R0052 and B. longum R0175 also attenuated the decremental effect of LPS on memory through brain-derived neurotrophic factor (BDNF) expression at the molecular level; however, this effect was not significant in the passive avoidance test at the behavioral level. CONCLUSIONS: These results suggest that the management of gut microbiota with this probiotic formulation could be a promising intervention to improve neuroinflammation-associated disorders such as AD.

Molecular identification and phylogenetic analysis of human Trichostrongylus species from an endemic area of Iran.

Human infections with Trichostrongylus species have been reported in most parts of Iran. The aim of this study was the identification, molecular characterization and phylogenetic analysis of human Trichostrongylus species based on ITS2 region of ribosomal DNA from Guilan Province, northern Iran. Stool samples were collected from rural inhabitants and examined by formalin-ether concentration and agar plate culture techniques. After anthelmintic treatment, male adult worms were collected from five infected cases. Genomic DNA was extracted from one male worm of each species in every treated individual and one filariform larva isolated from each case. PCR amplification of ITS2-rDNA region was performed and the products were sequenced. Among 1508 individuals, 46 (3.05%) were found infected with Trichostrongylus species using parasitological methods. Male worms of T. colubriformis, T. vitrinus and T. longispicularis were expelled from five patients after treatment. Out of 41 filariform larvae, 40 were T. colubriformis, and the other one was T. axei. Phylogenetic analysis showed that each species was placed together with reference sequences submitted to GenBank database. Intra-species similarity for all species obtained in the current study was 100%. T. colubriformis was found to be probably the most common species in this region of Iran. For the first time, the authors of the present study report the occurrence of natural human infection by T. longispicularis in the world. Therefore, the number of Trichostrongylus species infecting human in Iran now increased to ten.

An association study between CHEK2 gene mutations and susceptibility to breast cancer.

CHEK2 gene is known as a tumor suppressor gene in breast cancer (BC), which plays a role in DNA repair. The germ line mutations in CEHK2 have been associated with different types of cancer. The present study was aimed at studying the association between CHEK2 mutations and BC. Peripheral blood was collected from patients into a test tube containing EDTA, and DNA was extracted from blood samples. Then, we analyzed mutations including 1100delc, IVS2+1>A, del5395bp, and I157T within CHEK2 gene in patients with BC and 100 normal healthy controls according to PCR-RFLP, allelic specific PCR, and multiplex-PCR. Although IVS2+1G>A mutation within CHEK2 gene was found in two BC patients, other defined mutants were not detected. For the first time, we identified CHEK2 IVS2+1G>A mutation, one out of four different CHEK2 alterations in two Iranian BC patients (2%). Also, our results showed that CHEK2 1100elC, del5395bp, and I157T mutations are not associated with genetic susceptibility for BC among Iranian population.

Effect of calprotectin subunit S100A9 on the expression and methylation of OCLN in human melanoma cell line A-375.

Increased levels of calprotectin subunits S100A8 and S100A9 have been detected in human cancers. Melanoma is the most aggressive type of skin cancer, and its treatment is challenging because of its brain metastasis. OCLN encodes occluding, which plays a major role in the formation and regulation of tight junctions. The aim of this study was to evaluate the methylation status of the OCLN promoter and its expression in A-375 melanoma cells treated with or without various concentrations of S100A9 for 24, 48, and 72 h. Total RNA was extracted, and synthesized cDNA was assessed by performing real-time PCR. MSP-PCR was performed after treatment with bisultfie. Recombinant S100A9 inhibited the proliferation of the A-375 cell line and the expression of the OCLN gene was downregulated in a time- and concentration-dependent manner. Results of MSP-PCR showed that the OCLN gene promoter in a human melanoma cell line (A-375) was semimethylated.

The 763C>G Polymorphism of The Secretory PLA2IIa Gene Is Associated with Endometriosis in Iranian Women.

BACKGROUND: Endometriosis is a chronic gynecological disease resulting from complex interactions between genetic, hormonal, environmental and oxidative stress and intrinsic inflammatory components. The aim of this study was to investigate the potential association of the 763C>G polymorphism in the secretory phospholipase A2 group IIa gene (PLA2G2A) with the risk of endometriosis in Iranian women. MATERIALS AND METHODS: Ninety seven patients with endometriosis along with 107 women who were negative for endometriosis after laparoscopy and laparatomy, and served as the control group, were enrolled for this cross-sectional study. Samples were genotyped using the polymerase chain reaction-restriction fragment length polymorphism method. RESULTS: Multivariate analysis was used to examine the association between the risk of endometriosis and the 763C>G polymorphism of PLA2G2A. Genotype distributions of PLA2G2A were significantly different between patients and the controls (p<0.001, OR=0.22, 95% CI=0.21-0.39). Correlation analysis showed that there was a significant association between the normal homozygous genotype and susceptibility to endometriosis (p<0.001). CONCLUSION: The present study suggests that the 763C>G polymorphism of PLA2G2A plays an important role as an independent factor in the risk of endometriosis in Iranian women.