Cadherin 5 and Annexin V as Circulating Endothelial Microparticles: Markers for Atherosclerotic Vascular Lesions in Patients with Chronic Renal Failure

Cardiovascular disease is the leading cause of death in Chronic kidney disease patients. This study tries to identify circulating endothelial microparticles {MPs} [such as Cadherin 5 and Anexin V] in CKD patients with and without IHD as potential new risk factors of atherosclerotic vascular disease. This study was carried out in Theodor Bilharz Research Institute [TBRI] on 60 patients with chronic kidney disease on maintenance hemodialysis. They were 41 male and 19 females selected from hemodialysis unit in TBRI. They were further subclassified into the following two groups according to the Echocardiography and Electrocardiogram (ECG) to 25 patients of chronic kidney disease without cardiac complications (17 males, 8 females and ages were 53.5 ± 9.9 years) and 35 patients of chronic kidney disease with cardiac complications (24 males, 11 females and ages were 57.5 ± 7.4 years). Twenty healthy subjects were selected as healthy control, their age 50 ± 9 years. Cadherin 5 & Annexin V Were done by enzyme linked immunosorbant assay (ELISA). The mean cadherin 5 levels in CKD with ischemic HD, CKD without ischemic HD and control group were 86.99 ± 21.51, 33.21 ± 8.65 and 2.63 ± 1.47 respectively which significantly higher in CKD with ischemic HD and CKD without ischemic HD than control group (p < 0.01) and significantly higher in CKD with ischemic HD than CKD without ischemic HD (p < 0.01). As regard to the mean annexin v levels in CKD with ischemic HD, CKD without ischemic HD and control group were 83.73 ± 22.64, 28.51 ± 9.73 and 0.47 ± 0.36 respectively which significantly higher in CKD with ischemic HD and CKD without ischemic HD than control group (p < 0.01) and significantly higher in CKD with ischemic HD than CKD without ischemic HD (p < 0.01). Endothelial dysfunction leading to atherosclerotic vascular disease in patients with CKD can be assessed quantitatively by measurement of plasma levels of endothelial microparticles such as CD144-EMP (Cadherin 5) and Annexin V.     

How to protect liver graft with nitric oxide

Organ preservation and ischemia reperfusion injury as- sociated with liver transplantation play an important role in the induction of graft injury. One of the earliest events associated with the reperfusion injury is en- dothelial cell dysfunction. It is generally accepted that endothelial nitric oxide synthase (e-NOS) is cell-pro- tective by mediating vasodilatation, whereas inducible nitric oxide synthase mediates liver graft injury after transplantation. We conducted a critical review of the literature e...     

Method for efficient transfection of in vitro-transcribed mRNA into SK-N-AS and HEK293 cells: difference in the toxicity of nuclear EGFP compared to cytoplasmic EGFP

Here we report a method for efficient transfection of in vitro-transcribed mRNA into two different types of human adherent cells, the neuroblastoma cell line SK-N-AS, and the transformed kidney cell line HEK293. By using newly trypsinized adherent cells in suspension and Lipofectaminetrade mark 2000, we detected a transfection efficiency of 80-90% in both cell lines and a cell viability of 90% in SK-N-AS and 60% in HEK293, 24 h after transfection when using cytoplasmic enhanced green fluorescent protein (EGFP)-mRNA. We have evaluated the different effects of the generally used EGFP that mainly localizes to the cytoplasm and nuclear EGFP, where the nuclear EGFP are more toxic to the cells than the cytoplasmic EGFP. In order to develop a null experiment, we constructed a short non-functional mRNA including a nuclear localization signal and evaluated the concentrations at which mRNA encoding nuclear proteins can be added without a general toxicity, depending on the fact that the proteins are localized to the nucleus. For both SK-N-AS and HEK293 cells, a concentration of up to 100 ng mRNA in 10(5) cells, encoding a nuclear protein with no other function, did not affect the cells. For evaluation of the method, we screened four different human mRNAs, PDG, DFFA, CORT and PEX14, for their ability to affect cell proliferation in these cells. PEX14 was the only gene that significantly (p=0.03) reduced cell proliferation for both cell types, DFFA significantly (p=0.04) reduced cell proliferation in SK-N-AS but not in HEK293 cells. PGD and CORT did not have any effect on cell proliferation. We have developed an easy method for efficient delivery of in vitro-transcribed mRNA into the adherent cell lines, SK-N-AS and HEK293. This method is useful for a quick screening of how different genes affect cell proliferation.     

Long-term rugby practice enhances bone mass and metabolism in relation with physical fitness and playing position

We attempted to clarify whether rugby was able to enhance bone mineral density (BMD) and content (BMC) of weight-bearing bones by modifying bone remodelling in relation to physical fitness in male rugby players compared to controls. 22 male international rugby players aged 20–31 years (forwards, n = 12 and backs, n = 10) were compared to 14 age-matched controls of the same ethnic origin aged 21–33 years. BMD and BMC were measured by DEXA and bone metabolism was evaluated by specific makers. Physical fitness level was analysed in parallel. Both forwards and backs exhibited significantly higher BMD (+15%, +6%, respectively) and BMC (+36%, +18%, respectively) for whole body and all examined sites, regions and segments than controls, with more pronounced difference at forwards. The difference between both groups of rugby players and controls noted for BMD became more pronounced for BMC (+52% for lower limbs). BMD and BMC values were higher in forwards than backs at total body (+9, +16%) and at stressed sites such as spine (+12%, +13%), pelvis (+10%, +11%), upper (+17% for BMC) and lower limbs (+9%, +18%). Athletes, especially forwards, presented higher concentrations of OC and CTX resulting in positive values of the uncoupling index. Total body lean mass as well as muscular strength strongly correlated with total and stressed regional sites such as spine, pelvis, and lower limbs BMD and BMC (r = 0.59, p < 0.01 to r = 0.84, p < 0.001). Similarly, OC, but not CTX, is highly correlated with the same regional and total BMD and BMC (r = 0.67 to r = 0.81, p < 0.001). Rugby participation is associated with improved physical fitness, enhanced axial and appendicular bone mass and increased bone turnover in adult athletes. Moreover, the positive bone adaptation of these athletes may be related to an increase of bone remodelling rate in favour of bone formation, especially in forwards.     

Correlation of Resistin with Inflammatory and Cardiometabolic Markers in Obese Adolescents with and without Metabolic Syndrome

ObjectiveThe link between plasma resistin and obesity-related cardiometabolic disorders in children remains debatable. This study assessed the relationships of plasma resistin with cardiovascular risk factors, pro-inflammatory markers and insulin resistance index (HOMA-IR) in obese (Ob) adolescents and obese adolescents with metabolic syndrome (Ob-MS) compared to healthy controls (CO).Methods114 obese adolescents (60 Ob, age 13.6 ± 0.9 years, BMI 28.0 ± 2.2 kg/m², and 54 Ob-MS, age 13.8 ± 1.0 years, BMI 32.5 ± 4.8 kg/m²) and 37 CO (age 13.7 ± 0.8 years, BMI 22.8 ± 0.8 kg/m²) were studied. Anthropometrics, cardiac variables as well as fasting plasma concentrations of lipids, glucose, insulin, and adipocytokines (resistin, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), C-reactive protein (CRP)) were measured. HOMA-IR was calculated, and the presence of MS was assessed.ResultsPlasma resistin was significantly higher in Ob-MS than in both Ob and CO and was correlated with anthropometric, cardiovascular, pro-inflammatory markers and several components of MS as was HOMA-IR in Ob and Ob-MS. With increasing the number of MS components, plasma resistin, pro-inflammatory markers, and HOMA-IR were also increased. Multiple regression models highlighted significant correlation between resistin and both HOMA-IR (r = 0.40, p < 0.05) and systolic blood pressure (r = 0.63, p < 0.01) in Ob-MS. Conclusion:These results support the hypothesis that there is an association between circulating resistin and childhood obesity-related inflammatory and cardiometabolic events.Key Words: Resistin, Metabolic syndrome, Insulin resistance, Childhood obesity, Inflammation     

Antioxidant status and circulating lipids are altered in human gestational diabetes and macrosomia

Fetuses from mothers with gestational diabetes are at increased risk of developing neonatal macrosomia and oxidative stress. We investigated the modulation of antioxidant status and circulating lipids in gestational diabetic mothers and their macrosomic babies and in healthy age-matched pregnant women and their newborns. The serum antioxidant status was assessed by employing anti-radical resistance kit (KRL; Kirial International SA, Couternon, France) and determining levels of vitamin A, C, and E and the activity of superoxide dismutase (SOD). Circulating serum lipids were quantified, and lipid peroxidation was measured as the concentrations of serum thiobarbituric acid-reactive substances (TBARS). As compared with non-diabetic mothers, gestational diabetic women exhibited decreased levels of vitamin E and enhanced concentrations of vitamin C without any changes in vitamin A. Vitamin A and C levels did not change in macrosomic babies except vitamin E whose levels were lower in these infants than in the newborns of non-diabetic mothers. Gestational diabetes mellitus (GDM) and macrosomia were also associated with impaired SOD activities and enhanced TBARS levels. Globally, total serum antioxidant defense status in diabetic mothers and their macrosomic babies was diminished as compared with control subjects. Triglyceride and cholesterol concentrations did not differ significantly between gestational diabetic and control mothers; however, macrosomia was associated with enhanced plasma cholesterol and triglyceride levels. These results suggest that human GDM and macrosomia are associated with downregulation of antioxidant status, and macrosomic infants also exhibit altered lipid metabolism.     

Unilateral pseudouveitis revealing a pancreatic neuroendocrine carcinoma: A case report

Neuroendocrine tumors are a heterogeneous group of tumors with a wide range of malignant potential that tend to have a relative prolonged course. These tumors infrequently metastasize to the orbit. To the best of our knowledge, ocular metastases from pancreatic neuroendocrine tumors (PNETs) have never been reported in the literature. We report the case of a 61‐year‐old man who presented with progressive deterioration of general condition with unilateral recurrent episodes of non‐granulomatous panuveitis of the left eye related to a choroidal metastasis. Radiological imaging and histopathological analyses led to the diagnosis of metastatic pancreatic neuroendocrine carcinoma as the primary tumor. Choroidal metastases from neuroendocrine tumors are extremely rare, but compromise patients’ well‐being because of visual impairment. Uncommonly, these metastases can be the first manifestation of unknown tumors, warranting further investigations to detect the primary cancer.