The Relation Between Host TLR9 -1486T/C, rs187084 Gene Polymorphisms and Helicobacter pylori cagA, sodB, hsp60, and vacA Virulence Genes among Gastric Cancer Patients.

To identify the associations between different genotypes of TLR9 -1486T/C (rs187084) with gastric cancer patients and reveal their relation to Helicobacter pylori virulence genes (cagA, sodB, hsp60 and vacA). Patients with gastric cancer were recruited to our study, diagnosed both endoscopically and histopathologically. H. pylori were isolated from gastric samples by culture and PCR amplification of the glmM gene. Virulence genes cagA, sodB, hsp60, and vacA were detected by multiplex PCR. Blood samples were used for genotyping of TLR9 -1486T/C (rs187084) by PCR-RFLP. Out of 132 patients with gastric cancer, 106 (80.3%) were positive for H. pylori. A similar number of healthy participants was recruited as controls. The prevalence of cagA, sodB, hsp60, and vacA genes among H. pylori was 90.6%, 70.8%, 83.0%, and 95.3%, respectively. The vacA gene alleles had a prevalence of 95.3% for vacAs1/s2, 52.8% for vacAm1, and 42.5% for vacAm2. The CC genotype of TLR9 -1486T/C had a significantly higher frequency in gastric cancer patients when compared to healthy participants (p = 0.045). Furthermore, the CC genotype demonstrated a significant association with H. pylori strains carrying sodB, hsp60, and vacAm1 virulence genes (p = 0.021, p = 0.049, and p = 0.048 respectively). Patients with CC genotype of TLR9 -1486T/C (rs187084) might be at higher risk for the development of gastric cancer, and its co-existence with H. pylori strains carrying sodB, hsp60, or vacAm1 virulence genes might have a synergistic effect in the development of gastric cancer. Further studies on a wider scale are recommended.

Quinolone-resistant uropathogenic E. coli: is there a relation between qnr genes, gyrA gene target site mutation and biofilm formation?

Introduction. The resistance to quinolone reported in uropathogenic Escherichia coli (UPEC) is commonly caused by mutations in the target site encoding genes such as the gyrA gene. Bacterial plasmids carrying resistance genes such as qnr genes can also transfer resistance. Biofilms produced by UPEC can further aid the development of resistant urinary tract infections (UTIs).Hypothesis. Biofilm production is associated with higher prevalence of quinolones resistance genetic determinants.Aim. To detect the prevalence of qnr genes and gyrA gene mutation among quinolone-resistant UPEC and to investigate the relation between these genetic resistance determinants and biofilm production.Methodology. Catheterized urine samples were collected from 420 patients with evidence of UTIs and processed using standard techniques. Isolated UPEC were screened for quinolone resistance using an antimicrobial susceptibility test. Biofilm production among quinolone-resistant isolates was detected using the tissue culture plate method. All quinolone-resistant isolates were screened for qnr genes (qnrA, qnrB and qnrS) by multiplex PCR and for gyrA gene mutation by PCR-RFLP.Results. Two hundred and sixty-four UPEC isolates were detected from 420 processed urine samples. Out of the identified 264 UPEC, 123 (46.6 %) isolates were found to be quinolone-resistant, showing resistance to 1 or more of the tested quinolones. Of the 123 quinolone-resistant UPEC detected, 71(57.7 %) were biofilm producers. The qnr genes were detected among 62.6 % of the quinolone-resistant UPEC, with an estimated prevalence of 22.8, 32.5 and 37.4 % for qnrA, qnrB and qnrS genes, respectively. Additionally, the gyrA gene mutation was identified among 53.7 % of the quinolone-resistant isolates. We reported a significant association between biofilm production and the presence of qnrA, qnrB and qnrS genes. Furthermore, the gyrA gene mutation was significantly associated with biofilm-producing isolates. The coexistence of qnr genes, gyrA gene mutation and biofilm production was demonstrated in almost 40 % of the quinolone-resistant isolates.Conclusions. A significantly higher prevalence of qnr genes (qnrA, qnrB and qnrS) as well as the gyrA gene mutation was found among biofilm-forming UPEC. The reported coexistence of these different resistance mechanisms could aggravate quinolone resistance. Therefore, monitoring of resistance mechanisms and a proper stewardship programme are necessary.

Intrauterine Bacterial Colonization and Endometrial MicroRNA-17-5p Levels in Association to Endometriosis: A Study in an Egyptian Population.

We aimed to study the relation between both bacterial colonization of the uterine endometrium & endometrial miR-17-5p levels and endometriosis, and then to evaluate endometrial miR-17-5p as a biomarker of endometriosis. A comparative observational study was carried over 51 endometriosis patients and 51 controls admitted into Obstetrics and Gynecology department, Mansoura Faculty of Medicine. Endometrial tissue samples were collected and aimed for bacterial culture and identification of resulting organisms besides estimation of tissue levels of microRNA-17-5p by quantitative real time PCR. G. vaginalis, S. agalactiae, S. aureus, Mobiluncus and E. coli were associated with endometriosis. MicroRNA-17-5p was up-regulated in endometriosis patients (P value was <0.0001*). Its sensitivity and specificity were 90% and 76.5%. MiR-17-5p showed higher results in culture positive than negative cases. On studying the relation between the positivity of endometrial tissue culture and miR-17-5p and so endometriosis, P value was <0.0001*. We concluded that G. vaginalis, S. agalactiae, S. aureus, Mobiluncus and E. coli were associated with development of endometriosis. Endometrial miR-17-5p was elevated in association to positive detection of bacterial species. MiR-17-5p might be a bio- marker of endometriosis. ABBREVIATIONS: CFU/ml: Colony Forming Unit per Milliliter; miR-17-5p: MicroRNA-17-5p; qRT PCR: Quantitative Real Time Polymerase Chain Reaction.

Serum CD64 and ascitic fluid calprotectin and microRNA-155 as potential biomarkers of spontaneous bacterial peritonitis.

AIM: Patients with ascites are at a higher risk for associated of on top bacterial infections with subsequent life-threatening complications. We aimed to evaluate CD64, calprotectin, and microRNA-155 (miR-155) levels as diagnostic markers of spontaneous bacterial peritonitis (SBP) and the effect of using more than one use on the same spot over their diagnostic efficiency. PATIENTS AND METHODS: An observational comparative study included 103 patients with ascites admitted to the Tropical Medicine Department, Mansoura University Hospital, Egypt, divided into two groups: case group (64 patients) with ascites with SBP and control group (39 patients) with decompensated cirrhotic non-SBP ascites. Twenty milliliters of ascetic fluid was obtained from all participants for bacterial culture, and assessment of calprotectin and miR-155, in addition to 2 ml blood for the CD64 marker expression assay by a flowcytometer. RESULTS: The sensitivity and specificity of CD64 expression assay were 95.3 and 92.3%, respectively, area under the curve (AUC)=0.93, whereas those of ascetic fluid calprotectin and miR-155 were 87.5 and 82.1%, AUC=0.90 and 95.3 and 97.4%, with AUC of 0.95. Combined blood CD64 and ascetic fluid calprotectin had a diagnostic accuracy of 0.988 for blood CD64 and ascetic fluid miR-155, AUC=0.991, and that for ascetic fluid calprotectin and miR-155 was 0.988. On using the three studied markers together, the diagnostic accuracy was the best recorded, AUC=0.994. P values were less than 0.001. CONCLUSION: CD64, calprotectin, and miR-155 were good diagnostic markers of SBP and on using this combination, greater efficiency in diagnosis was achieved.

Maternal Serum sEndoglin and Cell-Free Fetal DNA as Probable Markers of Preeclampsia: A Study in Single Center, Egypt.

Background: This study was conducted to compare the levels of maternal serum soluble endoglin (sEng) and cell-free fetal DNA (cffDNA) in pregnant females with PE to normotensive pregnant ones, together with relating these levels to preeclampsia (PE) severity and onset. Method of the study: It was a comparative study in Mansoura University Hospital, Egypt, to detect the levels of serum sEng by ELISA besides the levels of cffDNA by quantitative real-time polymerase chain reaction in 80 pregnant females suffering from PE in addition to 80 normotensive pregnant ones that were included as control. Results: Levels of serum sEng and cffDNA were higher in PE cases than control (p < 0.0001Ù­ both) and were significantly related to the severity of the disease. Levels were also higher in early than late onset PE (p < 0.003Ù­ and <0.002Ù­, respectively). Sensitivities, specificities, positive, and negative predictive values in addition to accuracy of serum sEng and cffDNA were 97.5%, 98.8%, 98.7%, 97.5%, and 98.1% and 97.5%, 93.8%, 94.0%, 97.4%, and 95.6%, respectively. Conclusion: Maternal serum sEng and cffDNA can be good markers for diagnosis of PE in Egyptian patients. They are positively related to the disease severity. Abbreviations: cffDNA; Cell-Free Fetal DNA, sEng; soluble Endoglin, PE; preeclampsia, qRT PCR; Quantitative real-time polymerase chain reaction.

High-risk and low-risk human papilloma virus in association to spontaneous preterm labor: a case-control study in a tertiary center, Egypt.

INTRODUCTION: This study aimed to detect the correlation between human papillomavirus (HPV) and spontaneous preterm labor in Egyptian women and its association to the human papilloma viral load and MPP2 gene expression. MATERIAL AND METHODS: We performed an observational comparative case-control study in Department of Obstetric and Gynecology, Mansoura University Hospitals over women presented with spontaneous preterm labor, besides females admitted for giving birth at full term to detect conserved sequence in HPV-L1 gene (GP5/GP6) followed by genotype detection of high- and low-risk HPVs with quantification of the viral load and the MMP2 gene expression using real-time polymerase chain reaction (PCR). RESULTS: The prevalence of HPV was 18.1% in preterm females, but only 4% in full-term women (p value = 0.019*). Twenty percent were PCR positive for HPV 16 and 40% for HPV 18 whereas none of the control was positive for any of the studied high-risk genotypes. Thirty percent were PCR positive for HPV 6 and 10% were positive for HPV 11. MMP2 gene expression was significantly higher in preterm than full term. Human papilloma viral load was found to be positively correlated to the rate of MMP2 expression and the gestational age was significantly related to the viral load and the rate of expression of MMP2 gene. CONCLUSION: Human pabilloma virus especially high-risk genotypes was correlated to spontaneous preterm labor in Egyptian females through increasing early expression of MMP2 gene. The time of occurrence of preterm labor was affected by the viral load and so the rate of expression of MMP2 gene.

IL12 Gene Polymorphism in Association with Hepatocellular Carcinoma in HCV-infected Egyptian Patients.

BACKGROUND: Hepatocellular carcinoma has been recorded the commonest cancer in Egypt. This increasing incidence may be attributed to the high prevalence of hepatitis C virus with its complications, so this study aimed to investigate the association between the potentially functional polymorphisms of IL12A and IL12B genes as a risk factor of development of hepatocellular carcinoma (HCC) on top of hepatitis C virus (HCV) infection in an Egyptian population. MATERIALS AND METHODS: We genotyped two loci of IL12 which were rs568408 (3'UTR G>A) for IL12A and rs3212227 (3'UTR A>C) for IL12B in 78 patients with HCC on top of chronic HCV infection. In addition, 64 cancer-free chronic HCV patients were studied, besides 92 healthy subjects who were included as control. RESULTS: Study of rs568408 (G>A) gene polymorphism showed that the A allele is higher while the G allele is lower in HCC cases than cancer-free chronic HCV patients (p = 0.006*). The A-containing genotypes AG and (AG+AA) were higher while the GG was lower (p = 0.009* and p = 0.005*), respectively. The study of the rs3212227 (A>C) polymorphism showed neither statistically significant differences between the C and A allele (p = 0.2) nor between CC, AC, or AC+CC in HCC cases and cancer-free chronic HCV patients (p = 0.7, p = 0.2, and p = 0.29), respectively. CONCLUSION: Our findings showed that IL12A rs568408 (G>A) polymorphism may contribute to the risk of HCC on top of chronic HCV infection, whereas that of IL12B rs3212227 (A>C) do not.

UreA and cagA genes of Helicobacter pylori in Egyptian patients with laryngeal squamous cell carcinoma and benign laryngeal polyps: a cohort study.

This work aims to estimate the prevalence of Helicobacter pylori ureA gene and evaluate cagA gene-positive strains in both patients of laryngeal squamous cell carcinoma (LSCC) and those with benign laryngeal polyps. This study included 49 patients confirmed pathologically to have LSCC and 15 patients with benign laryngeal polyps over a period from June 2013 to March 2015. Samples of laryngeal tissue were collected during direct laryngoscope under general anesthesia to be pathologically evaluated followed by analysis for H. pylori detection. Each laryngeal tissue sample was divided into three parts; one for bacteriological examination, the second for pathological examination and the third for PCR to detect both ureA and cagA genes. Out of 49 LSCC samples, 31 (64.6 %) was positive for ureA by PCR. Out of them, 29 samples (93.5 %) were cagA positive. Only three cases (20 %) of the benign laryngeal polyp were ureA positive by PCR and one of them was cagA positive by PCR. By the bacteriological culture, only eight samples (25.8 %) gave growth. All of them were ureA positive and only seven of them were cagA positive. There was a significant association between presence of H. pylori and LSCC as compared to benign laryngeal polyp which may contribute in the pathogenesis of laryngeal carcinoma. These results should be confirmed by further studies over larger number of cases.

Interleukin-6, intracellular adhesion molecule-1, and glycodelin A levels in serum and peritoneal fluid as biomarkers for endometriosis.

OBJECTIVE: To compare levels of interleukin-6 (IL-6), intracellular adhesion molecule-1 (ICAM-1), and glycodelin A in serum and peritoneal fluid of patients with and without endometriosis, and to correlate levels with disease stage. METHODS: An observational study was undertaken at Mansoura University Hospital, Egypt, between March 2014 and June 2015. Patients aged 21-48 years laparoscopically diagnosed with endometriosis and those without endometriosis who underwent laparoscopy for tubal ligation were included. Levels of IL-6, ICAM-1, and glycodelin A were measured in samples of serum and peritoneal fluid. Receiver operating characteristic curves were used to evaluate diagnostic accuracy. RESULTS: Forty-eight women with endometriosis and 20 without the disorder were included. IL-6 and glycodelin A levels in serum and peritoneal fluid were higher in the endometriosis group than in the control group (P<0.001 for all); ICAM-1 levels did not differ. The sensitivity and specificity values were 93.8% and 80.0% for serum IL-6, 58.3% and 60.0% for serum ICAM-1, and 91.7% and 75.0% for serum glycodelin A. The corresponding values for peritoneal fluid markers were 85.4% and 89.0%, 60.4% and 50.0%, and 89.6% and 90.0%, respectively. IL-6 and glycodelin A levels in serum and peritoneal fluid increased with disease stage (P<0.001 for all). CONCLUSION: IL-6 and glycodelin A, but not ICAM-1, are potential biomarkers for endometriosis and are positively correlated with the disease stage.

Helicobacter pylori, Chlamydiae pneumoniae and trachomatis as probable etiological agents of preeclampsia.

OBJECTIVES: The aim of this study was to evaluate the evidence for Helicobacter pylori, Chlamydiae pneumoniae and trachomatis to act as a probable etiology for preeclampsia (PE), together with estimating the prevalence of such infections in pregnant women with PE. METHODS: We performed a prospective study in Mansoura University Hospitals, Egypt, for detecting H. pylori infection by estimating H. pylori IgG and IgM, in addition to detection of Chlamydiae infections by PCR in 90 pregnant women with PE and 90 normotensive pregnant women of the same age and body mass index who were studied as control. RESULTS: The prevalence of H. pylori infection in preeclamptic pregnant women was 54.4% with a statistically significant association to PE. The prevalence of C. pneumonia was 27.8% whereas that of C. trachomatis was 4.44%. The infected preeclamptic cases showed high levels of leucocytes besides elevated C-reactive protein concentrations. CONCLUSION: Helicobacter pylori was found to act as a cofactor in the development of PE. Occurrence of C. trachomatis was low in pregnant women in our community; however, it showed that it may act as a cofactor in PE, whereas C. pneumoniae was attributed to have no role in PE pathogenesis until supported by further studies.