Molecular characteristics of extended-spectrum beta-lactamases among gram-negative isolates collected in Cairo University Hospital

Phenotyping is commonly used for detection of extended-spectrum beta-lactamase (ESBL) production in gram-negative isolates. ESBLs are mainly coded for by four important genes, namely bla (TEM), bla (SHV), bla (CTX-M), and bla (OXA). Our aim in this study is to assess use of a multiplex PCR as a rapid method to identify four common genes responsible for ESBL production in different gram-negative isolates. All 793 clinical isolates are subjected to both screen and confirmatory testing for ESBL production using double disc synergy testing (DDST). Two hundred isolates with the ESBL phenotype are subjected to multiplex PCR for detection of the four genes bla (TEM, SHV, CTX-M, and OXA). The isolates were obtained from various clinical specimens: 68 (34 %) were isolated from urine cultures, 43 (21.5 %) from sputum, 26 (13 %) from wounds, 34 (17 %) from blood culture, 20 (10 %) from stool of healthy carrier and nine (4.5 %) from bronchoalveolar lavages. In this study, 83 isolates (41.5 %) were from outpatients (urine and stool specimens only), and the remaining 117 isolates (58.5) were from inpatients. By PCR technique, 181 isolates were found to be ESBL producers. blaTEM was the commonest genotype (39.2 %), followed by blaSHV (32.5 %) and blaCTX-M (30.9 %), either alone or in combination. Acinetobacter baumannii isolate had none of the ESBL genes. Eighteen (9.9 %) out of 181 isolates carried more than one type of beta-lactamase genes. Our study demonstrated rapid detection of bla (TEM, SHV, CTX-M, and OXA) in isolates belonging to Enterobacteriaceae and other nonfermenting clinical isolates using multiplex PCR. This genotypic method provided a rapid and efficient differentiation of ESBLs in the laboratory.     

Serum neutrophil gelatinase-associated lipocalin and cystatin C as markers of renal function in patients with stages 2–5 chronic kidney disease

Individuals with reduced glomerular filtration rate (GFR) are at greater risk for cardiovascular disease and other comorbidities. Creatinine-based equations are used to estimate GFR, identify patients with potential kidney disease, and classify them into different stages since serum creatinine is insensitive to changes in the GFR. The aim of our work was to evaluate diagnostic performance of serum cystatin C and neutrophil gelatinase-associated lipocalin (NGAL) as markers of kidney function in patients with reduced GFR. Fifty cases at different stages of renal impairment and 30 healthy control subjects were tested. Only serum NGAL and cystatin C were higher in stage 2 chronic kidney disease (CKD) when compared to the control group (p?<?0.05). For stages 3–5 the median levels of cystatin C, NGAL, and creatinine were found to be significantly higher than the control group. ROC curve constructed to differentiate stage 2 patients from the controls showed AUC for NGAL 0.795, sensitivity 86%, and specificity 63.3%; AUC for cystatin C 0.957, sensitivity 86%, and specificity 90%; and AUC for creatinine 0.738. Frequency of cases that tested positive for NGAL and cystatin C in stage 2 was higher than those in control group (p?<?0.05) with an OR of 10.364 (95% CI 1.099–97.686) for NGAL and OR 54 (95% CI 4.7–613) for cystatin C. NGAL and cystatin C exhibited higher sensitivity than creatinine for diagnosis of stage 2 CKD. Their use as adjunctive diagnostic tools in patients with mildly reduced GFR may be justified on the long term to diagnose early renal insult.     

Molecular epidemiology of cutaneous leishmaniasis and heterogeneity of Leishmania major strains in Iran

Objective To determine the geographical distribution of Leishmania species causing cutaneous leishmaniasis (CL) and to study the genetic heterogeneity of Leishmania major isolates from different endemic areas of Iran. Methods A total of 341 isolates from lesions of patients living in 11 provinces of Iran were grown in culture medium and inoculated to BALB/c mice to detect possible visceralisation. The species were identified by isoenzyme analysis using a battery of six enzymes and kinetoplast (k) DNA‐PCR technique. Genetic variation among L. major isolates was analysed by random amplified polymorphic DNA (RAPD) technique. Results Of the total 341 isolates, 283 isolates were L. major and 58 isolates were Leishmania tropica. In rural areas, the causative agent of CL was mainly L. major (95%L. major vs. 5%L. tropica), in urban areas it was L. tropica (65%L. tropica vs. 35%L. major). All isolates of L. major and 8.6% of L. tropica isolates showed visceralisation in BALB/c mice. There is considerable genetic diversity between L. major strains from different endemic areas and even between some isolates of the same endemic area. Conclusion Leishmania major is the most frequent species in the endemic areas of CL in eleven provinces of Iran, and genetic diversity is a common feature of L. major in the country.     

Pathogenesis of severe asthma

Patients with severe asthma have asthma symptoms which are difficult to control, require high dosages of medication, and continue to experience persistent symptoms, asthma exacerbations or airflow obstruction. Epidemiological and clinical evidences point to the fact that severe asthma is not a single phenotype. Cluster analyses have identified subclasses of severe asthma using parameters such as patient characteristics, and cytokine profiles have also been useful in classifying moderate and severe asthma. The IL-4/IL-13 signalling pathway accounts for the symptoms experienced by a subset of severe asthmatics with allergen-associated symptoms and high serum immunoglobulin E (IgE) levels, and these patients are generally responsive to anti-IgE treatment. The IL-5/IL-33 signalling pathway is likely to play a key role in the disease pathogenesis of those who are resistant to high doses of inhaled corticosteroid but responsive to systemic corticosteroids and anti-IL5 therapy. The IL-17 signalling pathway is thought to contribute to 'neutrophilic asthma'. Although traditionally viewed as players in the defence mechanism against viral and intracellular bacterial infection, mounting evidence supports a role for Th1 cytokines such as IL-18 and IFN-γ in severe asthma pathogenesis. Furthermore, these cytokine signalling pathways interact to contribute to the spectrum of clinical pathological outcomes in severe asthma. To date, glucocorticoids are the most effective anti-asthma drugs available, yet severe asthma patients are typically resistant to the effects of glucocorticoids. Glucocorticoid receptor dysfunction and histone deacetylase activity reduction are likely to contribute to glucocorticoid resistance in severe asthma patients. This review discusses recent development in different cytokine signalling pathways, their interactions and steroid resistance, in the context of severe asthma pathogenesis.     

Comparison between Gallbladder Serosal and Mucosal Patch in Duodenal Injuries Repair in Dogs

Background: One of the most challenging problems in clinical surgery is management of an extensive duodenal injury. In its management, there are limitations in using jejunal serosal patch and other conventional methods in specific conditions. This study was performed to compare treatment of large duodenal defects by a gallbladder serosal patch and the gallbladder mucosal patch in a dog as an animal model. Methods: A duodenal defect (2 cm, about 50% of the total circumference) was created in the second portion of the duodenum in eight dogs. The animals were divided into two equal groups, with group 1 undergoing serosal patch repair and group 2 undergoing mucosal patch repair. The macroscopic and microscopic healing features of the gallbladder serosal and mucosal patch were compared. Results: None of the dogs died due to surgical complications. The whole grafted area was covered by neomucosa at the end of the third week in all animals with the gallbladder serosal patch (group 1). In this group, the scar was small; no significant narrowing of lumen was noted and serosal healing was uniformly complete. In histological examination, a complete coverage of the gallbladder serosal patch by neomucosa consisting of columnar epithelium with short villous formations was observed. In mucosal patch models (group 2), complete epitheliazation, mild fibrosis, and incomplete repair were visible. In histological examination, severe inflammation was noticed too. Conclusion: In patients with multiple trauma affecting upper gastrointestinal tracts, use of the gallbladder serosal patch method is easy and reliable. So it may be considered in the surgical management of large duodenal defects, which cannot be repaired by available conventional methods.