Emergence of extensive drug resistance and high prevalence of multidrug resistance among clinical Proteus mirabilis isolates in Egypt.

BACKGROUND: Proteus mirabilis is an opportunistic pathogen that has been held responsible for numerous nosocomial and community-acquired infections which are difficult to be controlled because of its diverse antimicrobial resistance mechanisms. METHODS: Antimicrobial susceptibility patterns of P. mirabilis isolates collected from different clinical sources in Mansoura University Hospitals, Egypt was determined. Moreover, the underlying resistance mechanisms and genetic relatedness between isolates were investigated. RESULTS: Antimicrobial susceptibility testing indicated elevated levels of resistance to different classes of antimicrobials among the tested P. mirabilis clinical isolates (n = 66). ERIC-PCR showed great diversity among the tested isolates. Six isolates (9.1%) were XDR while all the remaining isolates were MDR. ESBLs and AmpCs were detected in 57.6% and 21.2% of the isolates, respectively, where blaTEM, blaSHV, blaCTX-M, blaCIT-M and blaAmpC were detected. Carbapenemases and MBLs were detected in 10.6 and 9.1% of the isolates, respectively, where blaOXA-48 and blaNDM-1 genes were detected. Quinolone resistant isolates (75.8%) harbored acc(6')-Ib-cr, qnrD, qnrA, and qnrS genes. Resistance to aminoglycosides, trimethoprim-sulfamethoxazole and chloramphenicol exceeded 80%. Fosfomycin was the most active drug against the tested isolates as only 22.7% were resistant. Class I or II integrons were detected in 86.4% of the isolates. Among class I integron positive isolates, four different gene cassette arrays (dfrA17- aadA5, aadB-aadA2, aadA2-lnuF, and dfrA14-arr-3-blaOXA-10-aadA15) and two gene cassettes (dfrA7 and aadA1) were detected. While class II integron positive isolates carried four different gene cassette arrays (dfrA1-sat1-aadA1, estXVr-sat2-aadA1, lnuF- dfrA1-aadA1, and dfrA1-sat2). CONCLUSION: P. Mirabilis ability to acquire resistance determinants via integrons may be held responsible for the elevated rates of antimicrobial resistance and emergence of XDR or even PDR strains limiting the available therapeutic options for management of infections caused by those strains.

Sulfadiazine analogs: anti-Toxoplasma in vitro study of sulfonamide triazoles.

Toxoplasmosis is an infection that prevails all over the world and is caused by the obligate intracellular protozoan parasite Toxoplasma gondii (T. gondii). Promising novel compounds for the treatment of T. gondii are introduced in the current investigation. In order to test their in vitro potency against T. gondii tachyzoites, six 1,2,3-triazoles-based sulfonamide scaffolds with terminal NH2 or OH group were prepared and investigated as sulfadiazine equivalents. When compared to sulfadiazine, which served as a positive control, hybrid molecules showed much more anti-Toxoplasma activity. The results showed that the IC50 of the examined compounds 3(a-f) were recoded as 0.07492 µM, 0.07455 µM, 0.0392 µM, 0.03124 µM, 0.0533 µM, and 0.01835 µM, respectively, while the sulfadiazine exhibited 0.1852 µM. The studied 1,2,3-triazole-sulfadrug molecular conjugates 3(a-f) revealed selectivity index of 10.4, 8.9, 25.4, 21, 8.3, and 29; respectively. The current study focused on the newly synthesized amino derivatives 3(d-f), as they contain the more potent amino groups which are recognized to be essential elements and promote better biological activity. Extracellular tachyzoites underwent striking morphological alterations after 2 h of treatment as seen by scanning electron microscopy (SEM). Additionally, the intracellular tachyzoite exposed to the newly synthesized amino derivatives 3(d-f) for a 24-h period of treatment revealed damaged and altered morphology by transmission electron microscopic (TEM) indicating cytopathic effects. Moreover, compound 3f underwent the most pronounced changes, indicating that it had the strongest activity against T. gondii.

Molecular and biological characterization of pyocyanin from clinical and environmental Pseudomonas aeruginosa.

BACKGROUND: Pyocyanin is a secondary metabolite secreted by P. aeruginosa. It is a redox-active blue/green phenazine pigment that has various beneficial applications. The present study aims at screening the production of pyocyanin among clinical and environmental P. aeruginosa isolates in Dakahlya governorate, Egypt. Thereafter, large-scale production, purification, structure elucidation, and assessment of the biological activity of the highest pyocyanin producers were targeted. RESULTS: Pyocyanin from the highest clinical (PsC05) and environmental (PsE02) producers were subjected to large-scale production, followed by purification using silica gel column. Pyocyanin was characterized using TLC, UV-Vis, 1 H NMR, and FTIR spectroscopy to confirm its structure and purity. Purified pyocyanin showed remarkable antimicrobial efficacy against all tested food-borne pathogens, MDR/XDR clinically isolated bacteria and C. albicans. Furthermore, it showed a substantial effect on biofilm inhibition and eradication of pre-formed biofilm against strong biofilm producing bacterial pathogens. However, it had limited antibiofilm activity against C. albicans. Pyocyanin from PsC05 had higher antioxidant and radicals scavenging activity than that from PsE02 as determined by FRAP, DPPH, and ABTS assays. Likewise, pyocyanin from PsC05 was more active against tested cancer cell lines, especially human Breast Cancer (MCF-7) and Colorectal Carcinoma (HCT-116), than that from PsE02. More importantly, it showed minimal cytotoxicity to normal cells. CONCLUSIONS: P. aeruginosa clinical and environmental isolates produce pyocyanin pigment in varying amounts. Pyocyanin exhibits substantial anti-bacterial, and anti-fungal activity; thus, enhancing its medical applicability. It could be used to inhibit and/or eradicate biofilm from the surfaces of medical devices which is a chief source of nosocomial infections. Its antioxidant along with cytotoxic activity against cancer cell lines, make it a promising contender for use as a substitute for synthetic agents in cancer treatment.

Nanoformulation-Based 1,2,3-Triazole Sulfonamides for Anti-Toxoplasma In Vitro Study.

Toxoplasma gondii is deemed a successful parasite worldwide with a wide range of hosts. Currently, a combination of pyrimethamine and sulfadiazine serves as the first-line treatment; however, these drugs have serious adverse effects. Therefore, it is imperative to focus on new therapies that produce the desired effect with the lowest possible dose. The designation and synthesis of sulfonamide-1,2,3-triazole hybrids (3a-c) were performed to create hybrid frameworks. The newly synthesized compounds were loaded on chitosan nanoparticles (CNPs) to form nanoformulations (3a.CNP, 3b.CNP, 3c.CNP) for further in vitro investigation as an anti-Toxoplasma treatment. The current study demonstrated that all examined compounds were active against T. gondii in vitro relative to the control drug, sulfadiazine. 3c.CNP showed the best impact against T. gondii with the lowest IC50 value of 3.64 µg/mL. Using light microscopy, it was found that Vero cells treated with the three nanoformulae showed remarkable morphological improvement, and tachyzoites were rarely seen in the treated cells. Moreover, scanning and transmission electron microscopic studies confirmed the efficacy of the prepared nanoformulae on the parasites. All of them caused parasite ultrastructural damage and altered morphology, suggesting a cytopathic effect and hence confirming their promising anti-Toxoplasma activity.

Evaluation of the effect of moringa oleifera gel and autologo platelet-rich fibrin in the treatment of rabbit intra bony defects. (Radio graphic and Histological study).

Purpose: Periodontitis is the most common condition, which causes bony defects; the ultimate goal of periodontal therapy is the regeneration of the destroyed tissues. There is always a need to search for better biomaterials that can be used for the treatment of intrabony defects. This study evaluated the effect of Moringa oleifera (MO) gel and platelet-rich fibrin (PRF) in the treatment of bone defects. Hypothesis: We hypothesized that MO gel may increase the bone mineral contents and density of bone. Methods: The study was conducted on 16 defects in 8 adult male rabbits divided into 2 groups; group (1) buccal bone defect treated with moringa hydrogel and PRF (right site), group (2) buccal bone defect treated with PRF (left site). Computed tomography (CT) radiography and histological examination were assessed at baseline, 14 and 28 days. The defects were induced in the form of one osseous wall defect between the 1st and the 2nd molars. Comparisons between groups were done using an unpaired t-test. For comparison within each group, analysis of variance (ANOVA) was used. Results: CT radiograph results showed there was a significant increase in bone density at 28 days in group 1 than in group 2 (843.13 ± 97.82 to 713.0 ± 51.09). The histological result revealed the defect area on the (PRF + Moringa) was almost filled completely by newly formed bone with few spots of retarded calcification. While (PRF) showed complete filling of the defect area by more fibrous tissue. The healing score showed a significant elevation of bone defect healing score in (PRF + Moringa group) when compared to (PRF group) at both times of evaluation. Conclusion: Radiographical examination, and histological and healing scores confirmed the superiority of Moringa + PRF results in an increase in bone fill and density in induced periodontal intrabony defects regeneration. Clinical trials should be considered to detect the effectiveness of MO in intrabony defects.

Inhibition of Erythromycin and Erythromycin-Induced Resistance among Staphylococcus aureus Clinical Isolates.

The increasing incidence of erythromycin and erythromycin-induced resistance to clindamycin among Staphylococcus aureus (S. aureus) is a serious problem. Patients infected with inducible resistance phenotypes may fail to respond to clindamycin. This study aimed to identify the prevalence of erythromycin and erythromycin-induced resistance and assess for potential inhibitors. A total of 99 isolates were purified from various clinical sources. Phenotypic detection of macrolide-lincosamide-streptogramin B (MLSB)-resistance phenotypes was performed by D-test. MLSB-resistance genes were identified using PCR. Different compounds were tested for their effects on erythromycin and inducible clindamycin resistance by broth microdilution and checkerboard microdilution methods. The obtained data were evaluated using docking analysis. Ninety-one isolates were S. aureus. The prevalence of constitutive MLSB, inducible MLSB, and macrolide-streptogramin (MS) phenotypes was 39.6%, 14.3%, and 2.2%, respectively. Genes including ermC, ermA, ermB, msrA, msrB, lnuA, and mphC were found in 82.6%, 5.8%, 7.7%, 3.8%, 3.8%, 13.5%, and 3.8% of isolates, respectively. Erythromycin resistance was significantly reduced by doxorubicin, neomycin, and omeprazole. Quinine, ketoprofen, and fosfomycin combated and reversed erythromycin/clindamycin-induced resistance. This study highlighted the significance of managing antibiotic resistance and overcoming clindamycin treatment failure. Doxorubicin, neomycin, omeprazole, quinine, ketoprofen, and fosfomycin could be potential inhibitors of erythromycin and inducible clindamycin resistance.

A novel pentavalent vaccine candidate completely protects against Acinetobacter baumannii in a mouse model of peritonitis.

Acinetobacter baumannii is considered as one of the most virulent and infectious organisms that have an increased ability to both evade host immune response and resist various classes of antibiotics, leading to life-threatening infections. Multiple virulence factors have been implicated in the high prevalence rate of A. baumannii in hospitalized and immunocompromised patients. Moreover, improper use of antibiotics has led to the emergence of extensive drug-resistant strains that urgently require alternative strategies to control this superbug. Unfortunately, the availability of a licensed vaccine against A. baumannii infections is still challenged by the vast diversity among A. baumannii strains. Here, we report the development of a novel pentavalent vaccine candidate composed of two recombinant proteins (Wza and YiaD) and a pool of capsular polysaccharides isolated from 3 clinical isolates. We tested this new vaccine in vivo in a mouse model of peritonitis against the standard strain ATCC 19606 in addition to 3 clinical isolates of A. baumannii. Immunization with this vaccine completely protected the challenged mice with 100% survival rate in the case of all the tested bacteria. Further clinical studies are urgently needed to evaluate the efficacy and safety of this proprietary vaccine to protect patients from A. baumannii lethal infections. KEY POINTS: • Recombinant proteins pool (Wza and YiaD) immunization led to a synergistic immune response. • Capsular polysaccharides pool induced up to 90% protection of tested clinical isolates. • The pentavalent pool showed superiority with 100% survival of immunized mice.

Novel PCR detection of CRISPR/Cas systems in Pseudomonas aeruginosa and its correlation with antibiotic resistance.

CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated proteins) systems are considered as acquired immune mechanisms in Gram-positive and Gram-negative bacteria and also in archaea. They provide resistance/immunity to attacking bacteriophages or mobile genetic elements as integrative conjugative elements (ICE) as well as plasmid transformation. As an opportunistic pathogen, Pseudomonas aeruginosa has been held responsible for serious infections especially in hospitalized and immunocompromised patients. Three subtypes of type I CRISPR system (I-C, I-E, & I-F1) have been detected in P. aeruginosa genomes. In this work, P. aeruginosa isolates were collected from different clinical sources, and the three CRISPR/Cas subtypes (I-C, I-E, & I-F1) were detected via singleplex and multiplex PCR techniques using novel universal primers that were designed specifically in this study. CRISPR subtypes I-C, I-E, and I-F1 were detected in 10, 9, and 13 isolates, respectively. Furthermore, antimicrobial susceptibility of CRISPR/Cas-positive and negative isolates to different antibiotics and the capacity of biofilm formation were detected using disc diffusion method and tissue culture plate method, respectively. There was a significant correlation between the presence/absence of CRISPR/Cas system and both antimicrobial susceptibility to some antibiotics and biofilm-forming capacity among P. aeruginosa clinical isolates. KEY POINTS: • A novel multiplex-PCR for detection of CRISPR/Cas-positive strains of P. aeruginosa. • Understand the correlation between CRISPR/Cas systems and other characters of P. aeruginosa. • Correlation between antimicrobial susceptibility and CRISPR systems in P. aeruginosa.

Nationwide hepatitis C virus screening and treatment of adolescents in Egyptian schools.

Until 2018, Egypt had the highest prevalence of hepatitis C virus (HCV) infection globally, affecting approximately 7% of the population. Despite efforts in diagnosis and treatment since 2006, nearly 2 million individuals with chronic HCV infection had yet to be diagnosed as of early 2018. In December, 2018, a mass HCV screening campaign for adolescents aged 15-18 years was initiated. Among 3 024 325 adolescents screened, the HCV antibody seroprevalence was 11 477 (0·38%), of whom 8187 (78·7%) were HCV RNA-positive. Sustained virological response 12 weeks after completion of treatment (SVR12) was attained by 7327 (99·6%) adolescents with a fixed-dose combination of generic ledipasvir 90 mg plus sofosbuvir 400 mg. Although mass screening in this age group might not be regularly adopted by many health systems and its cost-effectiveness might be lower than the screening of adults and high-risk groups (eg, patients on haemodialysis, people who inject drugs), breaking the chain of transmission in younger populations should lead to a reduction in HCV incidence and complications, and hasten the elimination of the disease.

Characterization of virulence determinants and phylogenetic background of multiple and extensively drug resistant Escherichia coli isolated from different clinical sources in Egypt.

Escherichia coli is a multifaceted microbe since some are commensals, normally inhabiting the gut of both humans and animals while others are pathogenic responsible for a wide range of intestinal and extra-intestinal infections. It is one of the leading causes of septicemia, neonatal meningitis, urinary tract infections (UTIs), cystitis, pyelonephritis, and traveler's diarrhea. The present study aims to survey the distribution and unravel the association of phylotypes, virulence determinants, and antimicrobial resistance of E. coli isolated from different clinical sources in Mansoura hospitals, Egypt. One hundred and fifty E. coli isolates were collected from different clinical sources. Antimicrobial resistance profile, virulence determinants, and virulence encoding genes were detected. Moreover, phylogenetic and molecular typing using ERIC-PCR analysis was performed. Our results have revealed that phylogroup B2 (26.67%) with the greatest content in virulence traits was the most prevalent phylogenetic group. Different virulence profiles and varying incidence of virulence determinants were detected among tested isolates. High rates of resistance to different categories of antimicrobial agents, dramatic increase of MDR (92.67%), and emergence of XDR (4%) were detected. ERIC-PCR analysis revealed great diversity among tested isolates. There was no clustering of isolates according to resistance, virulence patterns, or phylotypes. Our research has demonstrated significant phylogenetic diversity of E. coli isolated from different clinical sources in Mansoura hospitals, Dakahlia governorate, Egypt. E. coli isolates are equipped with various virulence factors which contribute to their pathogenesis in human. The elevated rates of antimicrobial resistance and emergence of MDR and XDR mirror the trend detected globally in recent years. KEY POINTS: • Clinical E. coli isolates exhibited substantial molecular and phylogenetic diversity. • Elevated rates of antimicrobial resistance and emergence of XDR in pathogenic E. coli. • B2 Phylogroup with the highest VS was the most prevalent among pathogenic E. coli.

Emergence of multidrug resistance and extensive drug resistance among enterococcal clinical isolates in Egypt.

INTRODUCTION: Enterococci commonly inhabit the gastrointestinal tract of both human and animals; however, they have emerged as a leading cause of several infections with substantial morbidity and mortality. Their ability to acquire resistance combined with intrinsic resistance to various antimicrobials makes treatment of enterococcal infections challenging. MATERIALS AND METHODS: The aim of the study was to evaluate the antimicrobial resistance pattern, and assess the prevalence of multidrug resistance (MDR) and extensive drug resistance (XDR) among enterococcal isolates, collected from different clinical sources, in Mansoura University Hospitals, Egypt. RESULTS: Antibiotic sensitivity testing revealed elevated levels of resistance among enterococcal clinical isolates (N=103). All E. faecium (N=32) and 74.6% of E. faecalis isolates(N=71) were MDR, while two E. faecalis and four E. faecium isolates were XDR. High level gentamicin resistance was detected in 79.6%, most of them carried the aac(6')-Ie-aph(2'')-Ia gene. High level streptomycin resistance was seen in 36.9%, of which 52.6% carried the ant(6')-Ia gene. Resistance to macrolides and lincosamides were mediated by ermB (92.2%) and msrA/B (42.7%). tetK, tetL, andtetM genes were detected among tetracyclines resistant isolates. Resistance to vancomycin was detected in 15.5%, where vanB and vanC1 gene clusters were detected in VRE isolates. Ten isolates (9.7%) were resistant to linezolid, eight of which harbored the optrA gene. Vancomycin and linezolid resistant enterococci were more likely to exhibit strong/moderate biofilm formation than vancomycin and linezolid sensitive ones. CONCLUSION: Elevated levels of resistance to different classes of antimicrobial agents and emergence of MDR and XDR strains pose a major threat with limited therapeutic options for infections caused by this emerging pathogen.

Incidence of Virulence Determinants Among Enterococcal Clinical Isolates in Egypt and Its Association with Biofilm Formation.

Background: Although Enterococci compromise an essential part of normal gut microbiota of both animals and humans, they have emerged as a leading opportunistic pathogen causing infections. The pathogenesis of enterococci is attributed to an array of virulence determinants. Objectives: This study aims to explore the prevalence and characteristics of enterococcal clinical isolates collected from Mansoura University Hospitals, Egypt, assess their ability to form biofilm, and the correlation with virulence determinants and antimicrobial resistance. Materials and Methods: A total of 70 Enterococcal clinical isolates were collected from different clinical sources between June and December 2016. Biofilm formation capacity was assessed, and characterization of virulence factors and antibiotic susceptibility was performed. Clonal relatedness between isolates was assessed using enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) approach. Results and Conclusion: The molecular analysis demonstrated high genetic diversity among enterococcal clinical isolates. The gelE was the most frequently detected gene (91.4%), followed by asa1 (70%), esp (65.7%), and cylA (17.1%), while hyl was not detected in any isolate. Gelatinase activity was detected in 35.7%, while hemolysin and lipase activity was detected in 12.9% and 78.5%, respectively. Most of the enterococcal isolates were biofilm producers, of which 67.1% were strong/moderate biofilm producers. All linezolid-resistant isolates exhibited strong/moderate biofilm formation capacity. Strong/moderate biofilm formation was more frequently observed among esp-positive (esp+) and gelatinase nonproducing (gelatinase-) enterococcal isolates. Multiple regression analysis denoted that esp (odds ratio [OR] 5.371, p = 0.003) and gelatinase production (OR 0.264, p = 0.015) were associated with strong/moderate biofilm formation capacity. These findings suggest that esp gene positivity and gelatinase production may affect biofilm formation capacity among enterococcal clinical isolates.

Prevalence and Mechanisms of Carbapenem Resistance Among Acinetobacter baumannii Clinical Isolates in Egypt.

The increasing number of carbapenem-resistant Acinetobacter baumannii clinical isolates is a major concern, which restricts therapeutic options for treatment of serious infections caused by this emerging pathogen. The aim of this work is to assess the antimicrobial resistance profile and identify the molecular mechanisms involved in carbapenem resistance in A. baumannii isolated from different clinical sources in Mansoura University Hospitals, Egypt. Antimicrobial susceptibility testing has shown that resistance to carbapenem has dramatically increased (98%) with concomitant elevated levels of resistance to quinolones, trimethoprim/sulfamethoxazole, and aminoglycosides. Polymyxin B and colistin are considered the last resort. Random amplified polymorphic DNA (RAPD) typing method revealed great diversity among A. baumannii isolates. Coexistence of diverse intrinsic and acquired carbapenem-hydrolyzing ß-lactamases has been detected in the tested isolates: Ambler class A: blaKPC (56%) and blaGES (48%), and Ambler class B: blaNDM (30%), blaSIM (28%), blaVIM (20%), and blaIMP (10%). Most isolates (94%) carried blaOXA-23-like and blaOXA-51-like simultaneously. blaOXA-23-like was preceded by ISAba1 providing a potent promoter activity for its expression. Sequencing analysis revealed that ISAba1 has been also inserted in carbapenem resistance-associated outer membrane protein (OMP) (carO) gene in three isolates, two of which were clonal based on RAPD typing, leading to interruption of its expression as confirmed by SDS-PAGE analysis of OMP fractions. Carbapenem resistance genes are widely distributed among A. baumannii clinical isolates from different clinical sources. Therefore, enhanced infection control measures, effective barriers, and rational use of antimicrobials should be enforced in hospitals for minimizing the widespread resistance to carbapenems and all other antibiotics.

Phenethyl isothiocyanate Triggers Apoptosis, Combats Oxidative Stress and Inhibits Growth of Ehrlich Ascites Carcinoma Mouse Model.

The aim of this study is to investigate the antitumor activity and possible molecular mechanism of Phenethyl isothiocyanate (PEITC) against Ehrlich ascites carcinoma in-vivo and in-vitro. In-vivo, ascetic fluid volume, body weight, serum malondialdehyde (MDA) level and total antioxidant capacity (TAC) were determined using Ehrlich ascites carcinoma (EAC) bearing mice. In-vitro, MTT assay was used. RT-PCR was used to investigate role of PEITC in apoptosis by analyzing the expression of Bax, caspase-9, and Bcl-2 genes. The effect of PEITC on caspase-9 enzyme activity was also tested. PEITC and/or Doxorubicin (Dox) treatment significantly suppressed EAC growth as compared to EAC/oil control mice. PEITC treatment showed a dose-dependent inhibition of EAC cells as indicated by MTT assay. We found that significant increase in MDA level and decrease in TAC caused by Dox treatment were significantly reduced by combination with PEITC treatment. Bax, caspase-9 genes' expression and caspase-9 enzymatic activity were significantly increased, while Bcl-2 gene expression was significantly decreased in PEITC treated mice. PEITC may act as a promising anticancer agent either alone or more effectively in combination with Dox through apoptotic cell death induction.

Co-production of AmpC and extended spectrum beta-lactamases in cephalosporin-resistant Acinetobacter baumannii in Egypt.

Acinetobacter baumannii is an opportunistic pathogen that has been held responsible for a lot of infections worldwide. Infections caused by this pathogen are difficult to control because of the widespread of antimicrobial resistance mechanisms. The aim of the present study is to assess the prevalence of extended spectrum ß-lactamases (ESBLs) and AmpC ß-lactamases among isolates of A. baumannii collected from different clinical sources in Mansoura University Hospitals, Egypt. Antimicrobial susceptibility testing has demonstrated elevated resistance level to ß-lactams, quinolones and aminoglycosides. All isolates were sensitive to colistin and polymyxin B. ESBL activity was detected in 86% of the isolates. Among the tested ESBL encoding genes, blaTEM gene was the most prevalent gene as it was detected in 52% of the isolates. While blaPER, blaSHV and blaVEB were detected in 12%, 4%, and 2%, respectively. AmpC activity and blaADC gene were detected in 90% of the tested isolates. Insertion sequence ISAba1 was located 9 bp upstream of blaADC gene in 88.9% of the ADC-expressing isolates providing a potent promoter activity for its expression. To our knowledge this is the first report of loss of intrinsic ADC activity, in 10% of the tested isolates, as a result of insertional inactivation by an element belonging to IS5 family transposase. Co-expression of both ESBLs and AmpC ß-lactamases was detected in 78% of the isolates. The study demonstrates high prevalence of resistance to ß-lactam antibiotics through ESBLs and AmpC ß-lactamases production among A. baumannii clinical isolates. Prevalence of ß-lactamases should be detected routinely and reported in hospitals to avoid inappropriate use of antibiotics and therapeutic failure.

A randomized controlled clinical trial of 4% sodium citrate versus heparin as locking solution for temporary dialysis catheters among hemodialysis patients
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BACKGROUND: Limited reports are available on the role of 4% citrate as a locking solution for temporary dialysis catheters. Hence, the aim of this study is to investigate the role of 4% citrate vs. heparin 5,000 µ/mL as a catheter-locking solution in a randomized controlled trial. MATERIALS AND METHODS: The trial was conducted in Egypt where the use of non-tunneled temporary catheters is prevalent compared to tunneled long-term catheters. The efficacy of catheter-locking solutions was compared regarding observation of rate of catheter dysfunction, low-flow pump, fever as a sign of central-line blood-stream infection (CLBSI), catheter-site infection, thrombosis, local bleeding, and systemic bleeding in each group of the study. RESULTS: Each group consisted of 105 patients. The number of patients who developed CLBSI in the citrate group was 11 (10.5%) compared to 23 (21.9%) in the heparin group (p < 0.025). The number of patients who developed catheter dysfunction in the citrate group was similar to those in the heparin group. The incidence of catheter-site infection, thrombosis, and local bleeding in the citrate group was similar to that in the heparin group. CONCLUSION: Citrate 4% lock solution is equally effective as heparin in maintaining catheter patency in dialysis patients. It may have a favorable effect on prevention of catheter-related infection due to its additional antiseptic properties as compared to heparin. Citrate-based locking solutions are a promising alternative to unfractionated heparin as a locking solution for dialysis catheters.
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Relationship of serum haemojuvelin and hepcidin levels with iron level and erythropoietin requirement in prevalent hepatitis C virus positive haemodialysis patients.

AIM: Iron overload is frequently reported in haemodialysis (HD) patients particularly those with chronic hepatitis C virus (HCV) infection. Soluble haemojuvelin (sHJV) has recently emerged as one of the significant regulators of iron homeostasis and hepcidin expression. The aim of the present study was to evaluate the potential associations of sHJV and hepcidin with inflammation, iron parameters and erythropoietin requirement in prevalent HD patients with HCV. METHODS: Serum sHJV and hepcidin were measured in 60 prevalent HD patients with [group I (n = 30)] and without [group II (n = 30)] HCV, and controls (n = 30) by enzyme-linked immunosorbent assay. Parameters related to anaemia, iron metabolism, inflammation, sHJV and hepcidin were measured. RESULTS: Serum hepcidin in HCV positive versus negative groups was 89.40 ± 46.08 ng/mL and 224.1 ± 72.36 ng/mL, P = 0.000, respectively, while sHJV was 245 ± 1.338 ng/mL and 254 ± 0.762 ng/mL, P = 0.147, respectively in positive versus negative patients. In group I, hepcidin correlated with serum ferritin (r = -0.512 P = 0.005) and transferrin saturation (TSAT%) (r = 0.572, P = 0.000) and sHJV correlated with ferritin (r = 0.40, P 0.000), TSAT% (r = 0.450, P = 0.002) and a significant correlation also existed between sHJV and hepcidin (r = -0.259, P = 0.045). In the regression analysis, ferritin and TSAT% were able to predict sHJV; (standardized ß = 0.52, P 0.001) and (standardized ß = 0.48, P 0.010). Ferritin and sHJV were also able to predict hepcidin (standardized ß = 0.627, P = 0.006) and (standardized ß = 0.300, P = 0.007) in group I. CONCLUSION: Soluble haemojuvelin levels seem to be associated with iron overload parameters and hepcidin levels in HCV positive HD patients.

Ectopic colonization of oral bacteria in the intestine drives TH1 cell induction and inflammation.

Intestinal colonization by bacteria of oral origin has been correlated with several negative health outcomes, including inflammatory bowel disease. However, a causal role of oral bacteria ectopically colonizing the intestine remains unclear. Using gnotobiotic techniques, we show that strains of Klebsiella spp. isolated from the salivary microbiota are strong inducers of T helper 1 (TH1) cells when they colonize in the gut. These Klebsiella strains are resistant to multiple antibiotics, tend to colonize when the intestinal microbiota is dysbiotic, and elicit a severe gut inflammation in the context of a genetically susceptible host. Our findings suggest that the oral cavity may serve as a reservoir for potential intestinal pathobionts that can exacerbate intestinal disease.

Central obesity and risks of cardiovascular events and mortality in prevalent hemodialysis patients.

BACKGROUND: To date, no attempt has been made to assess the best anthropometric method for defining abdominal adiposity in hemodialysis (HD) patients or to determine whether the quantity of intra-abdominal fat relates to morbidity and mortality in that population. We aimed to describe the prevalence of central obesity in HD patients and to investigate the relationship between central obesity assessed by anthropometric variables, and composite outcomes, cardiovascular morbidity and mortality among HD patients and whether this parameter correlates with intra-abdominal fat assessed by computed tomography scan (CT scan). METHODS: The procedures followed were in accord with the ethical standards of the committee on human experimentation of our institution. Informed oral consent was obtained from all patients. This was a cross-sectional study of 120 prevalent HD patients. Anthropometric measurements including body mass index, conicity index (Ci), waist-hip ratio (WHR), waist circumference (WC), waist-to-height ratio (WHtR), and visceral adiposity index (VAI) were recorded. Visceral and subcutaneous abdominal fat were assessed by CT scan. Comorbidity was scored for both the Charlson comorbidity index (CCI) and Davies comorbidity index. RESULTS: Twenty-eight patients (23.3%) were centrally obese based on anthropometry. By linear regression analysis, Ci, WHR, and VAI were predictors of CT assessed central obesity; p 0.042, 0.001, and 0.010, respectively. On assessment of the relationship between the abdominal obesity and the comorbidity indices, there was a positive significant correlation between Ci and CCI (p 0.025) and Davies score (p 0.002) which are predictors of mortality. During the mean follow-up period (3.2 years), 56 patients reached the composite outcome; eight patients died and 48 experienced CV events. Central obesity measured by anthropometry was a predictor of composite outcomes, cardiovascular morbidity, and mortality in HD patients by regression analysis and cox regression model. Only WC and WHtR did not predict mortality. CONCLUSION: Ci, WHR, and VAI are cheap alternatives for accurate assessment of morbidity and mortality risk in centrally obese prevalent HD patients.

Confounding effects of microbiome on the susceptibility of TNFSF15 to Crohn's disease in the Ryukyu Islands.

Crohn's disease (CD) involves chronic inflammation in the gastrointestinal tract due to dysregulation of the host immune response to the gut microbiome. Even though the host-microbiome interactions are likely contributors to the development of CD, a few studies have detected genetic variants that change bacterial compositions and increase CD risk. We focus on one of the well-replicated susceptible genes, tumor necrosis factor superfamily member 15 (TNFSF15), and apply statistical analyses for personal profiles of genotypes and salivary microbiota collected from CD cases and controls in the Ryukyu Islands, southernmost islands of the Japanese archipelago. Our association test confirmed the susceptibility of TNFSF15 in the Ryukyu Islands. We found that the recessive model was supported to fit the observed genotype frequency of risk alleles slightly better than the additive model, defining the genetic effect on CD if a pair of the chromosomes in an individual consists of all risk alleles. The combined analysis of haplotypes and salivary microbiome from a small set of samples showed a significant association of the genetic effect with the increase of Prevotella, which led to a significant increase of CD risk. However, the genetic effect on CD disappeared if the abundance of Prevotella was low, suggesting the genetic contribution to CD is conditionally independent given a fixed amount of Prevotella. Although our statistical power is limited due to the small sample size, these results support an idea that the genetic susceptibility of TNFSF15 to CD may be confounded, in part, by the increase of Prevotella.