Multiplex PCR for detection of bla CTX-M genes among the extended spectrum beta lactamase [ESBL] producing gram-negative isolates

Broad spectrum beta-Lactamase producing organisms are a growing world wide problem. Resistance has emerged ever to newer, more potent antimicrobial agents. Although there are several guidelines available for the phenotypic detection of ESBL producing bacteria. This remains a continuous issue. In this study, we used a multiplex PCR as a rapid method to identify bla CTX-M genes and discriminate between its groups that are responsible for ESBL production in members of Enterobacteriacae. Our study includes: 250 clinical isolates [23 sputum, 64 urine, 46 from blood, 28 from pus aspirates, 58 from entotracheal secretions, and 31 swabs from cellulitis, impetigo contagiosum [non bullous] and sycosis]. All isolates were biochemically identified, based on colony morphology, and was speciated by standard biochemical tests. ESBL enzyme production was confirmed by double disc synergy test according to CLSI guidelines. Multiplex PCR was performed for bla CTX-M of ESBL +ve isolates for detection and discrimination between groups. Our findings were as follows: out of 250 isolates; only 98 were proved to be resistant to different antibiotics by the disc diffusion method according to NCCLS: 3 of 53 [5.66%] Enterobacter. All from group [25/26]. 65 of 74 [87.8%] E.coli strains: -37 of which from groups [1] [CTX-M 15], 9 from group [1] [CTX-M-3], 8 from group [9] [CTX-M-14], 9 from group [9] [CTX M-9], 2 from group [25/26] [CTX-M 26]. 1 of 50 [2%] non fermenting gram -ve bacilli which is from group [25/26]. 29 of 73 Klebsiella strains [39.7%]: 19 from group [9] [CTX-M14] and 10 from group [9] [CTX-M 9]

Increased expression of soluble fractalkine [CX3CL1] in systemic sclerosis - possible role in vascular inflammation

The pathogenesis of systemic sclerosis [SSc] involves interplay between obliterative vasculopathy in multiple vascular beds, inflammation, autoimmunity and progressive fibrosis. Vascular injury and activation are the earliest and possibly primary events in the pathogenesis of SSc. To determine levels of serum soluble fractalkine [sFKN] and its receptor CX3CR1 in peripheral blood mononuclear cells [PBMCs] in systemic sclerosis [SSc] patients and healthy controls. In addition, to assess any possible association between sFKN and clinical features of SSc. Serum levels of soluble fractalkine [sFKN, CX3CL1] assessed by enzyme linked immunosorbent assay [ELISA] and expression of its receptor [CX3CR1] on peripheral blood mononuclear cells by flow cytometric analysis, were measured in 18 systemic sclerosis [SSc] patients and 15 age and sex matched healthy controls. The degree of skin involvement was estimated by modified Rodnan skin thickness score [mRSS], pulmonary involvement was assessed in all patients by high resolution computerized tomography [HRCT] and pulmonary function tests [PFTs]. Serum sFKN levels and expression of its receptor CX3CR1 were significantly elevated in SSc patients than in healthy controls [P < 0.0.05]. SSc patients with pulmonary fibrosis had sFKN levels three times higher than those without PF. Serum sFKN correlated inversely with forced vital capacity of lungs [FVC%] but correlated positively with severity of pulmonary fibrosis, extent of skin fibrosis [mRSS], pitting scars, skin ulcers, anti topo-isomerase 1 antibody and CRP. Serum sFKN may play an important role in the pathogenesis of SSc, including tissue inflammation and vascular injury, hence, its measurement may be a useful serologic marker for the diagnosis and follow up of pulmonary and skin complications. So strategies to target CX3CL1-CX3CR1 interaction could provide a new therapeutic approach in SSc, potentially by blocking endothelial cell injury, leucocyte

Exercise as a non pharmacological intervention and alternative to oral estrogen therapy in post menopausal women with endothelial dysfunction

Endothelial dysfunction is a key event in the progression of atherosclerosis and heart failure. When the vascular endothelium is healthy it become like Teflon and things don't stick but when it is unhealthy it become like a Velcro attracting blood born junk. Both exercise and postmenopausal estrogen therapy augments endothelial function through increasing bioavailability of nitric oxide [NO] which is a substance that keep your blood vessels opened. Determines the effects of acute bouts of exercise on brachial artery endothelium dependent flow mediated vasodilatation FMD in postmenopausal women. Whether these responses were augmented by the concurrent use of oral estrogen. Whether these two interventions independently or together achieve FMD values observed in pre menopausal women. This study was conducted on 30 apparently healthy post menopausal women their mean age was [54 +/- 4 years old]. FMD was quantified during supine rest and again 60 minutes after treadmill exercise for 45 minute at 60% v02 max - subjects were studied twice, before and after 4 weeks of oral estradiol. The normal reference values was obtained from concurrent determinant of FMD in 30 pre menopausal women their mean age was [28 +/- 2] years old under identical basal conditions. Flow mediated vasodilatation in post menopausal women markedly impaired when compared with pre menopausal women, the mean of absolute diameter change in brachial artery for flow mediated dilatation in post menopausal women was significantly less than premenopausal women [[2.01 +/- 0.2mm [6.1%] Vs 4.1 +/- 0.4mm [12%] P<0.05]. After exercise the absolute change in the brachial artery diameter for FMD in postmenopausal women significantly approximate normal values] [3.8 +/- 0.3mm [11.4%] P<0.05]. In contrast after estrogen therapy the mean of absolute brachial artery diameter change for FMD was augmented at rest [P < 0.01] but was not further enhanced after exercise. [3.7 +/- 0.6 mm [11.5%] VS 3.50 +/- 5mm [10.5%] P > 0.05]. Both interventions increased FMD to values in pre menopausal women. In post menopausal women both acute exercise and oral estrogen normalize FMD. However their effects weren't additive, so these results reinforce the concepts that exercise is an alternative non pharmacological intervention to estrogen in post menopausal women with endothelial dysfunction. For every post menopausal woman regular moderate intensity exercise training must continue to maintain improvement in your endothelial function

Mannose binding lectin: serum level, gene polymorphism in SLE patients and its relation to lupus nephritis

Systemic lupus erythematosus [SLE] is an autoimmune disease in which the complement system plays a crucial role in its pathogenesis. Mannan-binding lectin [MBL] is a recognition molecule of the lectin pathway of complement activation. The presence of several polymorphisms at the promoter and coding regions of the MBL-2 gene determines alterations at MBL serum concentration. MBL variant alleles that lead to low serum levels and/or functional deficits of MBL are postulated to contribute to the susceptibility of SLE. Moreover, the influence of MBL variation on antibodies production and renal involvement in SLE patients remains controversial. MBL serum level and genotypes were studied in SLE patients with evaluation of its role in auto antibodies production and lupus nephritis development. MBL genotypes and serum level were screened in a case control study included 30 SLE patients as well as 30 healthy controls. MBL polymorphism at exon 1 codons 54 and 57 was detected by PCR using sequence-specific priming [SSP] and serum MBL level was determined by ELISA technique. There was predominance of AA genotype [80%] in control group. Genotype frequencies of MBL variants in patients with SLE showed significant differences when compared with controls [AA 53.3% vs 80%, P=0.03, OR = 0.29 and A O+O O 46.6% vs 20%, P = 0.03, OR =3.5, respectively]. Serum MBL in SLE patients [900 ng/ml] was significantly lower than that of the control group [2750 ng/ml, P = 0.001] with positive correlation with low MBL genotypes. SLE patients with mutant alleles were more likely to produce anti dsDNA [92.8% vs 75%, OR = 4.3] and anti-Smith antibodies [35.7% vs 18.7%, OR = 2.3]. Patients carrying MBL-low genotypes have an increased risk of development of lupus nephritis than those carrying MBL-high genotypes [64.7% vs 35.2%, P - 0.02, OR= 2.4]. MBL gene polymorphism associated with low MBL serum levels that were found with significantly increased frequency in SLE patients may be one of the genetic factors that determine the susceptibility to develop lupus nephritis

Receptor activator of nuclear factor-kappa B ligand [RANKL] and osteoprotegrin [OPG] in rheumatoid arthritis

To evaluate serum and synovial fluid levels of receptor activator of nuclear factor Kappa B ligand [RANKL] and osteoprotegrin [OPG] in patients with rheumatoid arthritis [RA] and to determine the level of osteoclastic bone resorption by the ratio of RANKL to OPG as well as their correlation with the clinical activity, radiological grades and bone mineral density. 45 RA female patients and 15 age-matched healthy females with post traumatic knee effusion [as controls] were subjected to full history taking, complete clinical examination, assessment of disease activity using DAS score, radiological progression assessment using Larsen score, laboratory investigations including ESR, CRP and RF. Also, serum and synovial fluid levels of RANKL and OPG and bone mineral density [BMD] were done. RA patients had significantly higher levels of serum and synovial fluid RANKL, OPG and RANKL/OPG ratio than controls. Levels of RANKL, OPG and RANKL/OPG ratio were significantly higher in SF than in serum and in the active than in non active RA patients. The serum RANKL significantly correlated with disease duration, RF, ESR, DAS score and T- score, with no correlation with CRP or Larsen score. Synovial RANKL did not correlate with any of the previous parameters. As regards to serum OPG, it correlated with disease duration, RF, T-score and Larsen score, while synovial OPG showed correlation only with CRP. The serum RANKL/OPG ratio had significant positive correlation with CRP, DAS score and Larsen score. RANKL has a crucial role in the pathogenesis of bone loss in rheumatoid arthritis and can be used as a marker of bone erosion and disease activity. OPG plays a protective role against bone erosion and joint destruction in RA. RANKL/OPG ratio is more informative about bone resorption than each of RANKL or OPG alone

Clinical and surgical predictors of early post operative atrial fibrillation and sustained ventricular tachycardia after cardiac surgery

Post operative Atrial Fibrillation [AF]. Occurs up to 50% in cardiac surgery patients and represent the most common post operative complication. Although malignant ventricular tachycardia [V.T.] is uncommon arrhythmic complication early after cardiac surgery - it has a negative impact on mortality. The etiology of these arrhythmias [AF, V.T] after open heart surgery is incompletely understood and their prevention remains suboptimal. Identification of patients vulnerable for post operative [AF, V.T.] would allow targeting of these patients to benefit from aggressive prophylactic intervention. The aim of this work is to evaluate the incidence and identify risk factors of [AF, sustained V.T.] early postoperatively after cardiac surgery. 40 patients with a mean of age [55 +/- 10] years old [20 male, 20 female] under went isolated elective cardiac surgery [20 patients for valve replacement and 20 patients for CAPG]. Demographic and clinical data preoperative, operative and postoperative were collected. Patients continuously monitored and hemodynamically significant [AF, VT] were recorded. Detailed analysis was performed to define the risk factors. Post operatively AF occurred in [17/40] 42.5%. The mean age for patients with postoperative AF was 55 +/- 7.3 years old compared with 47.7 +/- 9.3 years old for patients without AF P<0.05. The mean heart rate variability [RMSSD] significantly differed between patients with post operative AF and patients without [15 +/- 2.1 msec VS 25 +/- 3 msec P<0.05]. The mean of P wave dispersion for patients with po AF was significantly prolonged compared to patients without AF [80 + 11 msec VS 42 +/- 12 msec, P<0.05]. Multivariate logesitic analysis [odds ratio +/- 95% CI, P value] was used to identify the following independent predictors of post operative AF: increasing age above VS below the mean age [OR = 2.8 CI [1.2-3.5] P<0.0] valve surgery VS CAPG [OR= 2.75 CI [1.2- 3.2] P<0.05], preoperative non use of beta blockers [OR= 1.5 CI [1.1-4.2] P<0.05] Considering several operative variables, use of internal mammary artery, pulmonary venting, cardiopulmonary bypass time, and aortic cross clamping time were significantly differed between the group with AF. And the group without AF. [26.6% VS 73.4% P=0.001] [71.4% versus 28.6% P=0.001], [113.8 +/- 33.5 m versus 92.4 +/- 36.3 m, P=0.002]. [97.8 +/- 21.5, versus 71.3 +/- 9.3, P = 0.001] respectively. Only one patient developed sustained VT post operatively [2.5%] of total study population, she was female had longer pump time than patient without sustained VT [120 min VS 80 +/- 9.5 min P<0.05]; longer Aortic cross clamping time [103 min VS 60 +/- 20 min P<0.05]; had increased QT[c]D than patient without sustained VT [120 msec VS 80 +/- 5 msec, P<0.05]. Patients with and without hemodynamically significant AF and sustained VT had similar body mass index preoperative heart rate and preoperative blood pressure. AF remains the most common complication after cardiac surgery. Increasing age and type of surgery identifies patients at risk for development of AF after cardiac surgery. Female sex, longer pump time, aortic cross clamping time, are independent predictors of developing sustanined VT post operatively. Increased QT[c] dispersion, decreased HRV, Root square of the mean of the sum of the Square of differences between adjacent R-R intervals [RMSSD] and increased PWD after cordic surgery may reflect disrupted electrophysiological stability of the myocardium and thus electrophysiological substrate for triggering malignant arrhythmias

Exercise as a non pharmacological intervention and alternative to oral estrogen therapy in post menopausal women with endothelial dysfunction

Both exercise and postmenopausal estrogen therapy augment endothelial function through increasing bioavailability of nitricoxide [NO]. The aim of this study was to: 1- determine the effects of acute bouts of exercise on brachial artery endothelium dependent flow mediated vasodilatation FMD in postmenopausal women. 2- Whether these responses were augmented by the concurrent use of oral estrogen. 3- Whether these two interventions independently or together achieve FMD values observed in pre menopausal women. This study was conducted on 30 apparently healthy post menopausal women their mean of age was [54 +/- 4 years old]. FMD was quantified during supine rest and again 60 minutes after treadmill exercise for 45 minute at 60% v[02] max - subjects were studied twice, before and after 4 weeks of oral estradiol. The normal reference values was obtained from concurrent determinant of FMD in 30 pre menopausal women their mean of age was [28 +/- 2] years old under identical basal conditions. flow mediated vasodilatation in post menopausal women markedly impaired when compared with pre menopausal women. The mean of absolute diameter change in brachial artery for flow mediated dilatation in post menopausal women was significantly less than premenopausal women [2.01 +/- 0.2mm [6.1%] Vs 4.1 +/- 0.4mm [12%] P<0.05]. After exercise the absolute change in the brachial artery diameter for FMD in postmenopausal women significantly approximate normal values [3.8 +/- 0.3mm [11.4%] P<0.05]. In contrast after estrogen therapy the mean of absolute brachial artery diameter change for FMD was augmented at rest [P < 0.01] but was not further enhanced after exercise. [3.7 +/- 1.32 mm [11.5%] VS 3.5 +/- 1.4mm [10.5%] P > 0.05]. Both interventions increased FMD to values in pre menopausal women. in post menopausal women both acute exercise and oral estrogen normalize FMD. However there effects weren't additive so these results reinforce that exercise is an alternative non pharmacological intervention to estrogen in post menopausal women with endothelial dysfunction

Methylene tetrahydrofolate reductase polymorphism as an atherosclerotic coronary risk factor among patients with coronary artery disease

Recently several studies revealed discrepancies about the association of methylenetetrahydrofolate reductase [MTHFR] gene polymorphism and the development atherosclerotic coronary artery disease [ACAD] in different races and population. The present study aimed to study MTHFR gene polymorphism among the controls and evaluate it as a risk factor for ACAD Egyptian patients. The study was conducted on 30 patients [58.7 +/- 6.02 years old]. Compared to age matehed 30 normal subjects. All patients underwent complete clinical ECG, enzymatic, assessment and coronary angiography fasting polymerase chain reaction-restricion fragment length polymorphism analysis was used to detect the C677T variants of the MTHFR gene in 30 patients with ACAD versus 30 healthy controls. Their serum homocysteine level was determined by ELISA technique. Fasling blood glucose, creatinine, cholesterol, high density lipoprotein cholesterol [HDL-C] and triglyceride were estimated by spectrophometric method, and low density lipoprotein cholesterol [LDL-C] was calculated. Frequencies of MTHFR CC, CT and TT genotypes were 50%, 30% and 20% in ACAD patient group versus 56.7%, 26.6% and 16.7% in control. Palients with ACAD showed significantly higher plasma homocyteine concentrations than control [12.03 +/- 3.4 imol/L VS 10.77 +/- 1.9 Rmol/L, P<0.05]. When palients and controls were classified according to their genotypes, no significant difference was found among the different genotypes in control group, as well as, between them and CC, CT type in patients group. In contrast, patients with MTHFR TT genotype had significantly higher homocysteine concentration compared to the same genotype in control group [17.01 +/- 3.4 Rmol/L VS 11.1 +/- 0.89 Rmol/L]. Furthermore, they had higher serum cholesterol and lower HDL-C, which have been correlated to their serum homocysteine concentrations. Relalive risk estimation revealed that palients with TT genotype were at greater risk than CT and CC genotypes. It could be concluded that MTHFR TT genotype could be considered as a risk factor that could interact with other environmental factors and contribute to ACAD