Investigation of the biofilm formation in extra-intestinal pathogenic Escherichia coli ST131 strains and its correlation with the presence of fimH, afa, and kpsMSTII genes.

Escherichia coli sequence type 131 (ST131) is a multidrug-resistant strain with the global dissemination. Biofilm formation-related factors include the most important virulence factors in extra-intestinal pathogenic E. coli (ExPEC) ST131 strains causing infections with treatment-limited subjects. This study aims to investigate the biofilm formation ability and its correlation with the presence of fimH, afa, and kpsMSTII genes in clinical isolates of ExPEC ST131. In this regard, the prevalence and characteristics of these strains collected and evaluated. The results revealed strong, moderate, and weak attachment abilities related to biofilm formation attributes in 45%, 20%, and 35% of strains, respectively. In the meantime, the frequency of the fimH, afa, and kpsMSTII genes among the isolates was observed as follows: fimH positive: 65%; afa positive: 55%; and kpsMSTII positive: 85%. The results convey a significant different of biofilm formation ability between clinical E. coli ST131 and non-ST131 isolates. Furthermore, while 45% of ST131 isolates produced strong biofilms, only 2% of non-ST131 isolates showed the ability to form strong biofilms. The attending of fimH, afa, and kpsMSTII genes in the majority of ST131 strains demonstrated a key role leading to biofilm formation. These findings suggested the application of fimH, afa, and kpsMSTII gene suppressors for treating biofilm infections caused by drug-resistant ST131 strains.

Prevalence of Listeria monocytogenes infection in women with spontaneous abortion, normal delivery, fertile and infertile.

BACKGROUND: Listeria monocytogenes with a vast range of natural reservoirs is more known for being a food-borne pathogen. Human infections have shown an impact on pregnancy outcomes, so, this study surveyed the frequency of L. monocytogenes infection involving different groups of women. METHODS: This study enrolled a total sample consisting of 109 women with spontaneous abortion, 109 women with normal delivery, 100 fertile women, and 99 infertile women aged 19-40 years and willing to participate in the study. The research tool in this study was a questionnaire and Polymerase chain reaction (PCR) test. RESULTS: According to the results, the frequency of L. monocytogenes infection was 4/109 (3.66%) observed among women with spontaneous abortion, 2/109 (1.83%) among women with normal delivery, 3/100 (3%) among fertile women, and 0/99 (0%) among infertile women. CONCLUSION: There was no significant relationship between Listeria monocytogenes infection and pregnancy outcomes of spontaneous abortion and infertility.

Virulence genes, phylogenetic analysis, and antimicrobial resistance of Escherichia coli isolated from urinary tract infection in hospitalized patients and outpatients.

Uropathogenic Escherichia coli (UPEC) strains are the most common cause of urinary tract infection (UTI) in hospitalized and community patients. The aim was to compare the genetic characteristics of E. coli isolated from inpatients (IPs) and outpatients (OPs) with UTI regarding their phylogenies, virulence traits, and resistance trends. In this cross-sectional study, 130 epidemiologically unrelated E. coli isolates were collected from patients with UTI. Extended-spectrum beta-lactamase (ESBL) production was detected by the combination disk method. UPEC and intestinal pathogenic E. coli (IPEC) virulence genes were detected by polymerase chain reaction. The isolates were analyzed for phylogenetic grouping. A P value of < 0.05 was considered significant. Of the 130 isolates, 62.3% were from OPs and 37.7% from IPs. About 35.8% of the OPs and 49% of the IPs were ESBL positive. Moreover, 56.8% of the OPs and 59.2% of the IPs were positive for UPEC virulence genes. Notably, 50% of the isolates from each group exhibited IPEC virulence properties. The predominant phylogroup was B2 (43.2% in the OPs and 40.8% in the IPs). No significant difference was found between the IP and OP isolates (P > 0.05). Our results may indicate that consideration should also be given to hygienic standards in the community. The marked genetic plasticity of E. coli has allowed the emergence of strains showing arrays of genes from different pathotypes. Characterization of E. coli isolates in different areas may guide the selection of effective infection control strategies.

Genotyping Helicobacter pylori and fgf7 gene expression in gastric cancer.

BACKGROUND: Helicobacter pylori as the causative agent of the most common chronic bacterial infectious disease in human still involves a range of clinical challenging complications. In this meantime, the survey of the interaction between H. pylori virulence genes expression and its consequences on gastric antral epithelial cells is Controversial. This study surveyed the correlations between H. pylori cag Pathogenicity Island and virulence factors genes with Fgf7 gene expression as an angiogenic factor in developing gastric cancer in gastric antral epithelial cells of patients with H. pylori infection. METHOD: Gastric antral biopsy samples collected from patients out of exclusion criteria, including consumption of tobacco, alchohol and anti-H. pylori drugs, were categorized into gastric cancer (case group n:53) and gastritis (control group n:50) with and without H. pylori infection to detect changes in cDNA of fgf7 in gastric antral epithelial cells by using Real Time RT PCR. Extracted total RNA from gastric antral biopsy samples was used to synthesize cDNA for real time PCR. Furthermore, the cDNA of H. pylori cag Pathogenicity Island and other virulence factors genes were detected by using specific designed primers and simple PCR. RESULTS: Fgf7 gene expression revealed a significantly increase in gastric antral epithelial cells of gastric cancer and H. pylori-positive patients in contrast with gastritis and H. pylori-negative patients (p < 0.05). In the meanwhile, cag Pathogenicity Island and hopQ genotypes showed a positive correlation with Fgf7 gene expression (fold changes of cDNA) in gastric antral epithelial cells (p < 0.05). CONCLUSION: This study revealed an obvious correlation between Fgf7 gene expression in gastric antral epithelial cells of patients with H. pylori carcinogenic genotypes infection and some host factors including age.

Sirt3, 6, and 7 Genes Expression in Gastric Antral Epithelial Cells of Patients with Helicobacter pylori Infection.

Sirtuins, known as the intracellular acylation enzymes, play a major role in regulating the cell's physiological activities. The relevant studies have shown diversely sirtuin genes expression in various cancers in humans. This study has surveyed the transcription of sirt3, 6, and 7 genes in gastric antral epithelial cells (GAECs) of gastritis and gastric adenocarcinoma patients with and without Helicobacter pylori infection. First of all, a case-control study was conducted, including 50 and 53 gastric antral biopsy samples collected from gastritis and gastric adenocarcinoma patients with and without H. pylori infection referred to hospitals of Sanandaj City during 2018-2019. Total RNAs were extracted from biopsy samples, then cDNAs were synthesized by using TaKaRa kits. Quality essay of H. pylori virulence genes expression and relative quantitative essay of sirt3, 6, and 7 genes expressions in gastric antral biopsy samples were performed using the real-time RT-PCR method. The statistical analysis showed the significant correlations between H. pylori vacA s1m2 and sabA cDNAs with sirt3 genes expression in GAECs (P < 0.05, 0.05 respectively). In addition, sirt6 gene's expression decreased along increasing age in gastric adenocarcinoma patients (P < 0.05). The samples of gastritis patients with gastric antral epithelial biopsy containing H. pylori hopQII, oipA, and sabB cDNA showed an increased amount of sirt7 genes expression (P < 0.05, 0.05, and 0.05 respectively). In conclusion, the H. pylori virulence genes expression and increasing age of patients showed the significant correlations with sirt3, 6, and 7 genes expressions in GAECs of gastric and gastric cancer patients.

Prevalence of Neisseria gonorrhoeae in Western Iran.

Neisseria gonorrhoeae is the causative agent of the sexually transmitted disease gonorrhoea. This bacterium infects the epithelial cells of the cervix of women and the urethra of men. However, its disease symptoms in the lower genitalia are found only in a small percentage of people. This study aimed to compare the frequency of N. gonorrhoeae genital infection among two groups of pregnant women, those with spontaneous abortions and those with normal pregnancies. This cross-sectional study was conducted in Western Iran. It included 417 women: 109 of whom had spontaneous abortions, 109 had normal deliveries, 100 were fertile, and 99 were infertile. Specific primers were used and DNA was extracted by endocervical swabs. A polymerase chain reaction test was then performed to detect N. gonorrhoeae. Data analysis was performed using the chi-squared test and t-tests. In all the above steps, a level of 5% was considered statistically significant, and the average ages in women with normal delivery, women with spontaneous abortion, fertile women, and infertile women were 27.8 ± 4.87, 29.6 ± 5.9, 32.1 ± 5.1, and 29.1 ± 6.3 years, respectively. The total frequency of N. gonorrhoeae infection was 0 (0%). The prevalence of N. gonorrhoeae infection was zero, and the disease was not associated with spontaneous abortion or infertility.

ZAK Gene Expression in Patients with Helicobacter pylori Infection.

BACKGROUND: ZAK protein is a member of the MLK family proteins defined as mediators in the cell cycle. A survey of ZAK gene expression in gastric antral epithelial cells (GAECs) of gastritis and gastric adenocarcinoma patients with Helicobacter pylori genotypes infection can elucidate carcinogenesis of H. pylori genotypes. METHODS: In a case-control study, ZAK gene expression was evaluated in GAECs biopsy samples of gastritis and gastric adenocarcinoma patients with (n 23, 21) and without H. pylori infection (n 27, 32), respectively. Total RNA was extracted from each gastric antral biopsy samples and cDNA synthesized by using Takara kits. H. pylori virulence genes֝ cDNA were detected by traditional PCR and specific primers. The ZAK gene expression was measured using the relative Real-Time RT PCR. RESULTS: The prevalence of gastric adenocarcinoma was the highest in man and 61-85 aged groups (p < .05). There was no significant correlation between the prevalence of H. pylori infection and patients' demographic groups. This study showed that ZAK gene overexpression gradually increases with increasing age and tumor grade among gastric adenocarcinoma patients. The gastric antral biopsy samples with H. pylori vacA s1m2 genotype infection showed a weak correlation with ZAK gene overexpression (p < .1). CONCLUSION: ZAK gene expression was higher in GAECs of gastritis cancer than in gastric adenocarcinoma, indicating the protective effect of ZAK against gastric cancer (p < .005). Reducing ZAK gene expression shows the negative correlations with H. pylori infection and gastric adenocarcinoma.

PREX2 gene's expression in gastric antral epithelial cells of patients with H. pylori infection.

BACKGROUND: The Prex2 protein is a member of the Rac family proteins that belongs to small G proteins with a critical role in cell migration, cell proliferation, and apoptosis through its effects on PI3K cell signaling pathway and phosphatase activity of PTEN protein. The effect of PREX2 gene expression has been shown in some cancer cells. A survey of PREX2 gene expression in gastric antral epithelial cells of gastric cancer patients with Helicobacter pylori various genotypes infection can conduct to better understanding H. pylori infection's carcinogenesis. METHODS: In a case-control study, PREX2 gene expression was evaluated in gastric antral biopsy samples on four groups of patients referred to Sanandaj hospitals, including gastritis with (n=23) and without (n=27) H. pylori infection and gastric cancer with (n=21) and without (n=32) H. pylori infection. Each gastric biopsy sample's total RNA was extracted and cDNA synthesized by using Kits (Takara Company). The PREX2 gene expression was measured using the relative quantitative real-time RT-PCR method and ΔΔCt formula. RESULTS: The PREX2 gene expression increased in gastric antral biopsy samples of gastritis and gastric cancer patients with H. pylori infection (case groups) than patients without H. pylori infection (control groups) 2.38 and 2.27 times, respectively. The patients with H. pylori vacA s1m1 and sabB genotypes infection showed a significant increase of PREX2 gene expression in gastric cancer antral epithelial cells. CONCLUSION: H. pylori vacA s1m1 and sabB genotypes have the positive correlations with PREX2 gene expression in gastric antral epithelial cells of gastritis and gastric cancer patients.

PREX2 gene's expression in gastric antral epithelial cells of patients with H. pylori infection / Expressão do gene PREX2 em células epiteliais antrais gástricas em pacientes com infecção por H. pylori

ABSTRACT BACKGROUND: The Prex2 protein is a member of the Rac family proteins that belongs to small G proteins with a critical role in cell migration, cell proliferation, and apoptosis through its effects on PI3K cell signaling pathway and phosphatase activity of PTEN protein. The effect of PREX2 gene expression has been shown in some cancer cells. A survey of PREX2 gene expression in gastric antral epithelial cells of gastric cancer patients with Helicobacter pylori various genotypes infection can conduct to better understanding H. pylori infection's carcinogenesis. METHODS: In a case-control study, PREX2 gene expression was evaluated in gastric antral biopsy samples on four groups of patients referred to Sanandaj hospitals, including gastritis with (n=23) and without (n=27) H. pylori infection and gastric cancer with (n=21) and without (n=32) H. pylori infection. Each gastric biopsy sample's total RNA was extracted and cDNA synthesized by using Kits (Takara Company). The PREX2 gene expression was measured using the relative quantitative real-time RT-PCR method and ΔΔCt formula. RESULTS: The PREX2 gene expression increased in gastric antral biopsy samples of gastritis and gastric cancer patients with H. pylori infection (case groups) than patients without H. pylori infection (control groups) 2.38 and 2.27 times, respectively. The patients with H. pylori vacA s1m1 and sabB genotypes infection showed a significant increase of PREX2 gene expression in gastric cancer antral epithelial cells. CONCLUSION: H. pylori vacA s1m1 and sabB genotypes have the positive correlations with PREX2 gene expression in gastric antral epithelial cells of gastritis and gastric cancer patients.

RESUMO CONTEXTO: A proteína Prex2 é membro das proteínas da família Rac que pertencem a pequenas proteínas G com um papel crítico na migração celular, na proliferação celular e na apoptose através de seus efeitos na via de sinalização celular PI3K e atividade fosfatase da proteína PTEN. O efeito da expressão genética PREX2 tem sido mostrada em algumas células cancerosas. Um levantamento da expressão genética PREX2 em células epiteliais antrais gástricas de pacientes infectados com vários genótipos de Helicobacter pylori pode conduzir a um melhor entendimento da carcinogênese da infecção por H. pylori. MÉTODOS: Em estudo de caso-controle, a expressão genética PREX2 foi avaliada em amostras de biópsia antral gástrica em quatro grupos de pacientes encaminhados aos hospitais de Sanandaj, incluindo gastrite com (n=23) e sem (n=27) infecção por H. pylori e de câncer gástrico com (n=21) e sem (n=32) infecção por H. pylori. O RNA total de cada amostra de biópsia gástrica foi extraído e cDNA sintetizado por meio de kits (Takara Company). A expressão genética PREX2 foi medida utilizando-se o método RT-PCR em tempo real quantitativo relativo e a fórmula ΔΔCt. RESULTADOS: A expressão genética PREX2 aumentou em amostras de biópsia antral gástrica de pacientes com gastrite e câncer gástrico com infecção por H. pylori (grupos de casos) em relação aos sem infecção por H. pylori (grupos de controle) 2,38 e 2,27 vezes, respectivamente. Os pacientes com infecção por genótipos H. pylori vacA s1m1 e sabB apresentaram um aumento significativo da expressão genética PREX2 em células epiteliais antrais de câncer gástrico. CONCLUSÃO: Os genótipos H. pylori vacA s1m1 e sabB têm correlações positivas com a expressão genética PREX2 em células epiteliais antrais gástricas de pacientes com câncer gástrico e gastrites.

Transcriptional Profile of Helicobacter pylori Virulence Genes in Patients with Gastritis and Gastric Cancer.

INTRODUCTION: Numerous molecular epidemiology studies have been performed about the frequency of Helicobacter pylori virulence genes in patients with H. pylori infection so far. This study was conducted to detect transcriptional profile by cDNA of H. pylori virulence genes in gastric biopsy samples of gastritis and gastric carcinoma patients. MATERIALS AND METHODS: In a case-control study, based on the prevalence of gastritis and gastric cancer in Sanandaj city during 2018 and 2019, 23 and 11 gastric antral biopsy samples with H. pylori infection were collected from gastritis and gastric carcinoma patients by the consecutive and available sampling method. Pathological characters, including tumor grades and tumor areas for gastric carcinoma biopsy samples prepared from gastric cancer areas, were determined by the pathologist. Total RNA of gastric antral biopsy samples was extracted, and their cDNA was synthesized by TaKaRa kit. H. pylori virulence genes' cDNA using specific primers and PCR was detected. This study's results were analyzed by SPSS version 25 and statics chi-square tests for determination of relationship and correlation between cDNAs of H. pylori transcriptional profile and clinical outcomes of H. pylori infection, including gastritis, gastric carcinoma, tumor grades, and tumor area. RESULTS: The positive statistical correlations were observed between transcripts of cagA, cagA-EPIYAC, cagE, and cagY genes and H. pylori infection clinical outcomes (P < 0.05). CONCLUSION: Detection of the H. pylori virulence genes' cDNA in gastric biopsy samples can help provide the prognosis of clinical outcomes.