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]

Diagnostic value of umbilical cord blood procalcitonin level as an early marker of neonatal sepsis

Septicemia is a systemic disease associated with the presence and persistence of bacteria and their toxins in the blood. It is one of the major causes of morbidity and mortality in the neonatal period. Early diagnosis and proper manag-ement of neonatal septicemia can bring down the morbidity and mortality rates. Procalcitonin [PCT] has recently become of interest and proposed as a possible marker of the systemic inflammatory response to infection in critically ill patients. The aim of this study was to clarify the diagnostic value of serum Procalcitonin [PCT] level as an early marker for diagnosis of neonatal sepsis. Twenty neonates with maternal risk factors for sepsis were included in this study. They included 8 males, 12 females with a mean gestational age of 38.2 +/- 1.76 weeks and mean birth weight of 3.25 +/- 0.34 kg. Twenty healthy children of matched age and sex with no maternal risk factors for sepsis were included in the study as a control group. Serum PCT concentrations were measured at umbilical cord blood samples, taken at time of delivery and peripheral blood samples taken 48 hours later using an immunochromatographic semi quantitative test [PCT-Q; Brahms, Hennigsdorf, Germany] and C- reactive protein [CRP] concentration was measured using an immune-oreactive quantitative method. Serum PCT concentrations were significantly higher in neonates with confirmed sepsis than both control group and neonates with clinically suspected but not confirmed sepsis. PCT is a more sensitive and specific early marker of neonatal sepsis compared with other laboratory parameters of sepsis

role of hepcidin level in iron overload in children with beta-thalassemia

Thalassemias are a group of inherited blood disorders with defective production of hemoglobin. Patients with beta-thalassemia develop iron overload due to increased iron absorption and transfusion therapy. Hepcidin is a hepatic hormone released in case of iron overload to regulate systemic iron homeostasis by inhibiting iron absorption from diet and recycling of iron by macrophages. To determine role of hepcidin in the pathogenesis of iron overload in 6-thalassemia. 20 Patients with beta thalassemia major [TM] included 10 males and 10 females, 20 patients with beta thalassemia intermedia [TI] included 10 males and 10 females and twenty healthy children of matched age and sex were included in this study. We assessed iron overload by measuring serum ferritin, assessed erythropoietic activity by measuring serum erythropoietin levels, and correlated these with urinary hepcidin measurements. We found severe urinary hepcidin deficiency in TI with strong inverse relationship between urinary hepcidin and serum erythropoietin levels in comparison with control group. In contrast, urinary hepcidin levels were elevated in TM with decrease of erythropoietin levels. In addition, serum ferritin level was significantly higher in TM than TI and significantly higher in TM and TI compared with normal control. Hepcidin deficiency may be the key factor allowing excessive iron absorption and iron overload in TI while in TM, chronic hemolysis and frequent blood transfusions may be the main factors that increase iron load

Study of soluble thrombomodulin as an early marker of endothelial cell injury in obese children

Obesity in children is associated with various cardiovascular risk factors as well as an increased cardiovascular disease morbidity and mortality in adulthood. Thrombomodulin, a glycoprotein with vasoprotective and anticoagulant properties, is located at the luminal endothelial cell surface. Its soluble form results from endothelial cell injury. The aim of this work was to study thrombomodulin as an early marker of endothelial cell injury in obese children. This study was done on 40 children: 20 obese and 20 non-obese control children. They were subjected to clinical examination and the following investigations, CBC, blood urea and serum creatinine, fasting blood glucose, serum cholesterol serum triglyceride, HDL-cholesterol, LDL- cholesterol and soluble thrombomodulin by ELISA. There was significant increase in blood pressure, fasting blood glucose, cholesterol, LDL-cholesterol, triglycerides and soluble thrombomodulin in obese children compared to control group [p < 0.05]. HDL-cholesterol was significantly lower in obese children compared to non-obese ones [p < 0.05]. There was significant positive correlation between blood pressure and plasma soluble thrombomodulin. From this study it could be concluded that obese children had more elevated blood pressure, fasting blood glucose and dyslipidemia. Soluble thrombomodulin was higher in obese children compared to controls. It was positively correlated with elevated blood pressure denoting endothelial cell injury in obese children

Medication non-compliance and oxidant-induced adverse effects of antiepileptic drugs in children

Poor compliance with prescribed medications and their side effects are significant health problems in chronic disease states as epilepsy. This work aimed to study medication non compliance of epileptic children and the possible role of free radical injury in antiepileptic drugs side effects. The study was done on eighty-two epileptic children subjected to history taking, clinical examination and they were given antiepileptic drugs with follow-up for one year. Thirty healthy, age and sex matched children were studied as a control group. Complete blood count, liver and renal function tests were performed for all patients at the beginning and the end of study. Therapeutic drug monitoring for antiepileptic drugs given was performed for all patients. Nitric oxide [NO], superoxide dismutase [SOD] and malondialdehyde [MDA] were measured for all patients who developed any drug side effects, thirty patients without drug side effects chosen randomly, and control group. Unsatisfactory compliance was reported in 51% of cases. Serum levels of antiepileptic drugs were not matched with seizure outcome. Complicated regimens were associated with unsatisfactory compliance [P<0.05]. NO, MDA and SOD were significantly higher in both patients groups receiving antiepileptic drugs with or without side effects compared to control group. Also, these parameters were significantly higher in patients who developed side effects compared to patients without side effects to antiepileptic drugs [P<0.05]. It could be concluded that simple antiepileptic drug regimens are needed, focusing on drug compliance is essential by depending not only on drug levels but also on regular follow-up of patients and good physician- patients relation. Role of oxidant injury in producing antiepileptic drugs side effects is suspected and should be confirmed by further studies

Prognostic value of serum S100 protein level in newborn infants with hypoxic ischemic encephalopathy

Hypoxic ischemic encephalopathy [HIE] is the most common cause of neurologic disease during neonatel period and is associated with high mortality and morbidity rate including cerebral palsy, mental retardation and seizures. S100 beta[2] is normally present in serum in very low concentrations, but found in high concentrations in the brain both in glial cells and in neurons. Serum S100 protein peaked in the first day after birth in asphyxiated newborn infants. The aim of this study was to clarify the prognostic value of serum S100 protein level as a marker of cerebral injury in newborn infants with birth asphyxia [HIE]. 20 newborns with HIE were investigated successively in the first 3 days after birth in comparison with 20 healthy newborn infants as a control group. S100 protein levels were detected by a monoclonal two-site immuno-luminometric assay. Follow up of the cases for 6 months after discharge from incubators was done to detect cases that developed cerebral palsy. We found significant increase in serum S100 protein level in newborn infants with birth asphyxia as compared to control group, also we found significant positive correlation between day 1 S100 protein levels and severity of HIE and positive correlation between day 1 S100 protein levels and future development of cerebral palsy. Early determination of serum S100 protein in first day after birth in newborn infants with hypoxic ischemic encephalopathy may be used as a good marker for assessment of severity of HIE and extent of brain damage and to predict the possibility of future development of cerebral palsy in newborn infants with HIE