Open-array analysis of genetic variants in Egyptian patients with type 2 diabetes and obesity

Background: Diabetes mellitus is considered a major public health problem worldwide. Susceptibility to diabetes is influenced by both genetic and environmental determinants

Aims/hypothesis: The aim of the present study was to test for 16 independent single nucleotide polymorphisms [SNPs] in established Type 2 diabetes [T2D] and obesity susceptibility loci by GWAS in a sample of Egyptian patients to find out if there is shared genetic background underlying both disease entities

Methods: Genotyping was performed using OpenArray protocol on the QuantStudio 12K Flex Real-Time PCR System. In the present case control study a custom array was designed to facilitate cost-effective analysis of selected SNPs related to glycolysis, gluconeogenesis, inflammation, insulin signalling, and immune function

Results: Seven gene variants showed significant association with the risk of T2D patients including FCGRA2, STAT4, CELSR2, PPARG, EXT2 rs3740878, GCKR, PTGS1. Factors that significantly affect T2D were obesity [p < 0.001] and GCKR [p = 0.001] and PTGS1 [p = 0.001] gene variants. Gene variants that showed significant or borderline effect on obesity were MTHFD1, EXT2 rs3740878, GCKR and PTGS1 [p = 0.03, 0.017, 0.059, 0.006] respectively

Conclusions/interpretation: Overlapping genetic aspects should be considered and the presence of risk alleles of different genes together could contribute to the risk of T2D or obesity or both. The MTHFD1 and EXT2rs3740878 gene variants significantly affect obesity and not shared with T2D. Gene variants that showed combined effect on both disease entities were GCKR and PTGS1. These findings provide a basis for future studies on a larger scale. More stress on the risk gene variants that have a combined impact on both diabetes and obesity is recommended to improve risk prediction and preventive strategies

Contribution of different mechanisms to the resistance to fluoroquinolones in clinical isolates of Salmonella enterica

OBJECTIVES: To study the potential factors include gene mutation, efflux pump and alteration of permeability associated with quinolone-resistance of Salmonella enterica strains isolated from patients with acute gastroenteritis and to evaluate the degree of synergistic activity of efflux pump inhibitors when combined with ciprofloxacin against resistant isolates. METHODS: Antimicrobial resistance patterns of fifty-eight Salmonella isolates were tested. Five isolates were selected to study the mechanism of resistance associated with quinolone group, including mutation in topoisomerase-encoding gene, altered cell permeability, and expression of an active efflux system. In addition, the combination between antibiotics and efflux pump inhibitors to overcome the microbial resistance was evaluated. RESULTS: Five Salmonella isolates totally resistant to all quinolones were studied. All isolates showed alterations in outer membrane proteins including disappearance of some or all of these proteins (Omp-A, Omp-C, Omp-D and Omp-F). Minimum inhibitory concentration values of ciprofloxacin were determined in the presence/absence of the efflux pump inhibitors: carbonyl cyanide m-chlorophenylhydrazone, norepinephrin and trimethoprim. Minimum inhibitory concentration values for two of the isolates were 2-4 fold lower with the addition of efflux pump inhibitors. All five Salmonella isolates were amplified for gyrA and parC genes and only two isolates were sequenced. S. Enteritidis 22 had double mutations at codon 83 and 87 in addition to three mutations at parC at codons 67, 76 and 80 whereas S. Typhimurium 57 had three mutations at codons 83, 87 and 119, but no mutations at parC. CONCLUSIONS: Efflux pump inhibitors may inhibit the major AcrAB-TolC in Salmonella efflux systems which are the major efflux pumps responsible for multidrug resistance in Gramnegative clinical isolates.

P53 Sensitizes human colon cancer cells to hesperidin through upregulation ofbaxand p21

Hesperidin, a flavonoid found mainly in citrus, was reported to inhibit growth and proliferation of several cancers, including colon cancer cells. However, the question does p53 tumor suppressor protein is required for the effect of hesperidin is not yet clarified. In the present study, the effect of hesperidin on p53-expressing [HCT116 p53[+/+]] and p53-knockout [HCT116 p53[+/+] human colon cancer cells was investigated. Hesperidin inhibits cell growth of both HCT116 p53[+/+] and HCT116 p53 [A]cells, however, it was more effective in p53-expressing cells. Hesperidin induced Gl cell cycle arrest in only HCT116 p53[++] cells however induction of reactive oxygen species [ROS] and apoptosis was induced in both cells. Furthermore, hesperidin activates the proapoptotic [Bax] and cyclin dependent kinase inhibitor [p21] in only HCT116 p53[+/+] cells. Interestingly, using p53 transcriptional inhibitor [pifithrin-alpha] hesperidin-inducing Bax and p21 upregulation in only HCT116 p53[+/+] cells was reduced by cotreatment with pifithrin-alpha without inducing any changes in HCT116 p53-/- cells. Altogether; these results showed that hesperidin induced apoptosis and Gl cell cycle arrest in colon cancer cells in a p53/Bax - dependent and- independent, and p53/p21 - dependentmanners; respectively

Efficacy of different antibiotic combinations on uropathogenic multi-drug resistant Escherichia coli

Using checkerboard titration method as well as time-kill curve technique, the potential synergy of combinations of beta-lactams [ceftazidime and cefepime], aminoglycosides [amikacin, gentamicin, and tobramycin] and fluoroquinolones [ciprofloxacin, levofloxacin, and ofloxacin] were compared against multidrug resistant uropathogenic Escherichia coli isolated from patients with urinary tract infections. In the checkerboard titration studies, none of 21 combinations demonstrated antagonism against 8 strains, five of the 21 combinations showed synergism for more than 40% of the test strains, i.e. ceftazidime tobramycin, ceftazidime levofloxacin, amikacin levofloxacin, amikacin with ofloxacin, and tobramycin with levofloxacin, synergy occurred more often with levofloxacin combined with amikacin [7/8 strains]. Corresponding to the respective fractional inhibitory concentration [FIC] indices, the bactericidal activity determined in combinations of amikacin with ceftazidime or levofloxacin at sub-minimum inhibitory concentrations produced a significant reduction in bacterial count [>/= 2 log 10 cfu/ml] observed within 6 hours of drug administration, at 24 hours, both combinations were bactericidal [reduction in colony count >/= 3 log 10]. Results of this study suggest that amirioglycosides [amikacin and or tobramycin] combination with levofloxacin or ceftazidime could be promising alternatives for the treatment of serious urinary tract infections due to multi drug resistant E. coli

antimicrobial activity of actinomycetes against multi-drug resistant bacteria

The study investigates the antibacterial and antifungal activity of some actinomycetes microorganisms. Twenty actinomycetes were isolated from different soils at various locations in Egypt. Cultural, morphological and physiological characteristic of this isolates indicated that they belong to Streptomyces sp. The isolates have antagonistic activity against different Gram negative and Gram positive multi-drug resistant bacteria and pathogenic fungi. Two isolates namely Streptomyces bobili FS-3 and Streptomyces poonensis FS-4 were further investigated. The potency of antimicrobial activity depend on incubation time, the nutritional factors such as carbon, nitrogen and phosphorus sources and environmental factors such a initial pH, medium capacity, size, type and age of inoculum

Single staged surgery [SSS] for left sided colonic obstruction [LSCO]

There are controversial opinions for management of left sided colonic obstruction. The orthodox surgeons prefer to do twice or triple sittings for the treatment of this situation. Because of the safety and avoid time consuming and cost-effectiveness, single staged colonic surgery is a safe and definitive procedure. From a period of four years duration between March 2000 to March 2004, 37 cases [22 men and 15 women] were under went to this single-typed technique for left sided colonic obstructive patients. We performed resection anastomosis [one layer] without fecal [bowel] diversion. The results were good and encouraging the surgeon to continue and perform this single-staged procedure for selected patients presented with left-sided colonic obstruction. This decreases the hospital stay, cost-effectiveness and reduce surgical intervention

Anaesthesia and intensive care for patients with morbid obesity

Fifty major surgical procedures were carried out on 48 morbidly obese patients in king Fahd Hospital -Jeddah and Jubail Hospital, Eastern Province, from year 1407 to year 1414. There were 44 females and 4 males. The age range was 14-52 Years [mean 31.8, S.D. 8.08]. The weights ranged from 97-183 kg [mean 126.97 kg, S.D 17.86]. All patients received general anaesthesia and they were all admitted to the Intensive Care Unit after operation. Fourteen patients needed prolonged endotracheal intubation with assisted ventilation after operation for an average of 9.7 hours. While the average stay in I. C. U. was 39.5 hours. No special anaesthesia tevhnique was recommended, the only emphasis was on careful monitoring during anaesthesia of all the vital signs, the arterial blood gases and acid base balance and on the necessity of having a central venous line and arterial line. In the I. C. U., hypoxaemnia occurred in 14 patients [simple in 9, severe and recurrent in 5]. Other metabolic changes included hyperglycaemia in [35 cases], hypocalcaemia [13 cases], elevation of the liver enzymes in [32 cases] and acidosis in [4 cases] We recommend post-operative extubation as early as possible and we reserve I. C. U. beds only for patients who have additional medical problems