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This work is investigating functional, histopathological, and molecular changes underlying Helicobacter pylori hypomotility and is correlating these with miR-1, whose disturbance is supposed to be involved in smooth muscle dysfunction and cell proliferation according to literature. Epithelial to mesenchymal transition and reduced ghrelin hormone may contribute to H. pylori infection-associated hypomotility. H. pylori infection was associated with reduced pyloric miR-1 expression. Targeting miR-1 could be valuable in the clinical management of pyloric hypofunction.

Multiple theories have been proposed describing the pathogenic mechanisms of

Mera Mohammed Galal Anan,1 Eman Ahmed El-Seidi,1 Marwa Salah Mostafa,1 Laila Ahmed Rashed,2 Doaa Mahdy El-Wakil1 1Department of Medical Microbiology and Immunology, Faculty of Medicine, Cairo University, Cairo, Egypt; 2Department of Biochemistry and Molecular Biology, Faculty of Medicine, Cairo University, Cairo, EgyptCorrespondence: Mera Mohammed Galal AnanDepartment of Medical Microbiology and Immunology, Faculty of Medicine, Cairo University, Kasr Al-Ainy St., Cairo, 11562, EgyptTel +20 1288866285Email mira.hassan36203@postgrad.kasralainy.edu.egPurpose: Colistin represents one of the last treatment options for infections caused by multi-drug resistant (MDR) Enterobacterales. The emergence of a plasmid-mediated mobile colistin resistance-1 (mcr-1) gene has raised serious concerns about its potential dissemination among bacteria.Methods: In this study, we evaluated the chromogenic medium, CHROMID® Colistin Resistance (COLR) agar, for the rapid detection of colistin-resistant Enterobacterales using broth microdilution (BMD) as a reference method. We also attempted to detect mcr-1, − 2, − 3, − 4, and − 5 genes, as well as the insertion sequence ISApl1 via polymerase chain reaction (PCR), followed by sequencing of mcr gene(s).Results: Among the 100 studied Enterobacterales isolates, 53% of them were colistin-resistant, with higher rate among Klebsiella pneumoniae (75%) as compared to Escherichia coli (44.4%). The COLR agar showed 83.2% sensitivity and 97.9% specificity for the detection of colistin resistance. Among colistin-resistant isolates, mcr-1 gene was only detected in four (7.5%) E. coli isolates. The ISApl1 was not found among mcr-1 positive isolates. Sequencing of mcr-1 gene revealed nucleotide sequence homogeneity with the wild-type mcr-1 gene in BLAST.Conclusion: The COLR agar is a promising phenotypic method for the detection of colistin-resistant Enterobacterales. Multiplex PCR followed by sequencing can be used for mcr genes’ detection and characterization.Keywords: colistin resistance, Enterobacterales, mcr genes, CHROMID® COLR agar, multiplex PCR, sequencing

Purpose Colistin represents one of the last treatment options for infections caused by multi-drug resistant (MDR) Enterobacterales. The emergence of a plasmid-mediated mobile colistin resistance-1 (mcr-1) gene has raised serious concerns about its potential dissemination among bacteria. Methods In this study, we evaluated the chromogenic medium, CHROMID® Colistin Resistance (COLR) agar, for the rapid detection of colistin-resistant Enterobacterales using broth microdilution (BMD) as a reference method. We also attempted to detect mcr-1, −2, −3, −4, and −5 genes, as well as the insertion sequence ISApl1 via polymerase chain reaction (PCR), followed by sequencing of mcr gene(s). Results Among the 100 studied Enterobacterales isolates, 53% of them were colistin-resistant, with higher rate among Klebsiella pneumoniae (75%) as compared to Escherichia coli (44.4%). The COLR agar showed 83.2% sensitivity and 97.9% specificity for the detection of colistin resistance. Among colistin-resistant isolates, mcr-1 gene was only detected in four (7.5%) E. coli isolates. The ISApl1 was not found among mcr-1 positive isolates. Sequencing of mcr-1 gene revealed nucleotide sequence homogeneity with the wild-type mcr-1 gene in BLAST. Conclusion The COLR agar is a promising phenotypic method for the detection of colistin-resistant Enterobacterales. Multiplex PCR followed by sequencing can be used for mcr genes’ detection and characterization.

Background Helicobacter pylori (H. pylori) infection has been recognized as a significant threat for gastric cancer. However, studies that investigated the oncogenic factors and antimicrobial resistance of H. pylori in Egyptian isolates with gastric cancer are rare. The current study aimed to examine: (1) The pattern of antimicrobial resistance of H. pylori isolates of Egyptian gastric cancer patients, and (2) the prevalence of Cytotoxin-associated gene A (CagA). Methods Samples were collected from patients with gastric cancer. Isolation of H. pylori was performed using Columbia blood agar supplemented with 10% horse blood, and selective supplement of H. pylori for 3 to 5 days at 37 °C under microaerophilic condition. Isolates were identified by biochemical traits of H. pylori: oxidase, urease and catalase tests. Antimicrobial susceptibility of H. pylori isolates was examined against five antimicrobial agents using disc diffusion method. After that, extraction of DNA and Polymerase Chain Reaction (PCR) were performed to amplify the target genes. Results Twelve samples were collected from six males and six females Egyptian patients with cancer with an age range from 22 to 65 years. These cases are scarce and samples were collected over a period of almost eleven months. All isolates were confirmed as positive H. pylori through colony morphology and biochemical tests. The most effective antibiotic found was ciprofloxacin whereas all isolates showed resistance to metronidazole and erythromycin. The target CagA oncogene gene with expected product size was reported and seven (out of twelve) isolates (58%) were identified as CagA positive. Conclusion The current study is unique in two main aspects. First, it reported the pattern of antimicrobial susceptibility and prevalence of CagA gene in H. pylori from Egyptian patients. Second, it exclusively recruited isolates from gastric cancer patients which were confirmed by clinical and laparoscopic examination. The moderately high prevalence of CagA gene in Egyptian cancer patients calls for more vigilance against that oncogene.

Background: Urine cultures constitute the majority of the workload for a microbiology laboratory with only 20%-30% of urine sample resulting in significant growth. Chromogenic media (CM) are available for urine specimens to enable rapid identification of common pathogens and also has been reported to increase mixed culture detection, reducing unnecessary workup. Chromogenic media offers the potential to lower costs by providing decreased work time, storage space and identification costs. Objectives: The present study focused on evaluation of the chromogenic medium (CPS) for the diagnosis of UTI in comparison with CLED as a conventional medium. Methodology: Over the period of January to July 2014, fifty urine samples with >100 pus cells /HPF were examined. CPS and CLED media were used for direct inoculation in addition to conventional biochemical reactions and/or API as needed. Results: In comparison with CLED, CPS showed a sensitivity of 93.5%, specificity of 100%, positive predictive value of 100%, negative predictive value of 57.1% and total agreement of 94%. The sensitivity of CPS for E. coli was 95%, for KESC was 75%, for Proteeae was 100% and for Enterococcus was 100%. The specificity of CPS for detection of E. coli was 100%, for KESC was 100%, for Proteeae was 97.9% and for Enterococcus was 100%. Conclusion: CPS proved to be a rapid, cost-effective diagnostic method for urinary tract infections. Therefore, CPS can replace the standard primary plating media used in routine diagnosis of urinary tract infection

The present study aimed to investigate the potential virulence factors and antifungal resistance of 31 Candida albicans and 21 non-albicans Candida isolates colonizing preterm neonates. The study also compared the susceptibility results with the in vitroactivity of chlorhexidine in the eradication of Candida colonization. Candida albicansproduced significantly more phospholipase and coagulase than non-albicans Candida, whereas proteinase production was higher in non-albicans Candida. Biofilm production was demonstrated in Candida albicans and non-albicans Candida (P = 0.214). None of the planktonic growth of Candida isolates were resistant to either fluconazole or amphotericin B, whereas 40% and 84% Candida isolates grown as biofilm became resistant to fluconazole and amphotericin B, respectively. Both coagulase and phospholipase production strongly correlated with the resistance of sessile Candidaisolates to amphotericin B (P < 0.001). Whereas both proteinase and phospholipase correlated with the resistance of in vitro Candida biofilms to fluconazole (P < 0.05 and P = 0.001; respectively). Chlorhexidine was comparable to fluconazole towards planktonic and sessile grown Candida isolates. In conclusion, the study demonstrated an association between certain virulence factors and the development of biofilm drug resistance and highlighted the value of chlorhexidine as a promising prospect in the eradication ofCandida colonization. Key words: Antifungal susceptibility, biofilm resistance, Candida colonization,chlorhexidine, preterm neonates, virulence factors.

Background: Estimated as the second or third most prevalent respiratory pathogen in the pediatric population, routine testing for human metapneumovirus (hMPV) can have a pivotal impact on children’s clinical outcome. Objectives: This cross-sectional analytical study aimed to determine the efficiency of direct fluorescent antibody (DFA) assay as a rapid tool for the diagnosis of hMPV infection as compared to real time reverse transcriptase polymerase chain reaction (rRT-PCR). In the meantime, we endeavored to analyze the clinical features in hMPV patients. Methods: A total of 50 children aged ≤ 24 months presenting with manifestations of acute respiratory tract infection (ARTI) at El-Mounira pediatric university hospital, Cairo university were enrolled in the study. Nasopharyngeal aspirates (or endotracheal aspirates in intubated children) were examined with the DFA assay as well as rRT-PCR as a gold standard for the detection and quantification of hMPV. Results and conclusion: Human MPV was detected in two cases by DFA and in four cases by rRT-PCR among hospitalized children with ARTIs. The DFA assay proved to be a highly specific test, yet with low sensitivity when compared to rRT-PCR. Most of hMPV-infected cases presented during the winter season, with January and February exhibiting the highest hMPV activity. Pneumonia was the most common presentation of ARTIs in hMPV-infected patients. Direct evaluation of respiratory specimens by DFA provides rapid results with low cost and a subsequent early medical management. However, its use should be restricted as a first-line approach, and a confirmatory test would be needed for a definite diagnosis.

Objective Over 50% of the world populations are infected with Helicobacter pylori (H. pylori). Most subjects are asymptomatic; however, in 1994, H. pylori has been categorized as group I carcinogen. The aim of the study was to investigate the relationship between H. pylori infection and gastric cancer.

Background and study aims Ascitic fluid infections (AFIs) are the frequent complications of advanced liver disease. Bacterial translocation is considered a key step in the pathogenesis of gut-derived bacterial infections; mainly spontaneous bacterial peritonitis (SBP) in cirrhotic patients. Bacterial DNA (bactDNA) in ascitic fluid and serum has been suggested as a surrogate marker for bacterial translocation. We attempted at the isolation and identification of bacteria in ascitic fluid in cirrhotic patients and the assessment of polymerase chain reaction (PCR) in ascitic fluid and serum. Patients and methods Fifty cirrhotic patients having ascites with no signs of infection were included. Ascitic fluid cultures were obtained from patients. Ascitic fluid and serum were subjected to DNA extraction and PCR for the universal amplification of a region of the 16S ribosomal RNA (16S rRNA) gene to detect bactDNA. Results Bacteria were isolated from 9 (18%) of the ascitic fluid samples, and were mainly Gram-positive bacteria. BactDNA was detected simultaneously in the ascitic fluid and serum of 17 (34%) patients and in the ascitic fluid of only 2 patients. In a single patient with positive ascitic fluid culture no bactDNA was detected in ascitic fluid or serum. By considering AFIs as a positive ascitic fluid culture and/or the presence of bactDNA in the ascitic fluid and/or serum, ascitic fluid culture could detect 9 out of 20 patients with AFIs (45%), PCR of ascitic fluid could detect 19 out of 20 (95%) while PCR of serum could detect 17 out of 20 (85%). In 10 patients with culture negative non-neutrocytic ascites (CNNNA) bactDNA could be detected in serum and ascitic fluid. Conclusion AFI can be caused by Gram positive as well as Gram negative organisms. A substantial percentage of cases with CNNNA show bactDNA in serum and ascitic fluid. PCR of ascitic fluid should, therefore, be used in the diagnostic workup of suspected cases of ascitic fluid infections.