BRITISH JOURNAL OF PHARMACOLOGY, Issue 3 2010
Morag K Mansley
BACKGROUND AND PURPOSE Insulin-induced Na+ retention in the distal nephron may contribute to the development of oedema/hypertension in patients with type 2 diabetes. This response to insulin is usually attributed to phosphatidylinositol-3-kinase (PI3K)/serum and glucocorticoid-inducible kinase 1 (SGK1) but a role for protein kinase B (PKB) has been proposed. The present study therefore aimed to clarify the way in which insulin can evoke Na+ retention. EXPERIMENTAL APPROACH We examined the effects of nominally selective inhibitors of PI3K (wortmannin, PI103, GDC-0941), SGK1 (GSK650394A) and PKB (Akti-1/2) on Na+ transport in hormone-deprived and insulin-stimulated cortical collecting duct (mpkCCD) cells, while PI3K, SGK1 and PKB activities were assayed by monitoring the phosphorylation of endogenous proteins. KEY RESULTS Wortmannin substantially inhibited basal Na+ transport whereas PI103 and GDC-0941 had only very small effects. However, these PI3K inhibitors all abolished insulin-induced Na+ absorption and inactivated PI3K, SGK1 and PKB fully. GSK650394A and Akti-1/2 also inhibited insulin-evoked Na+ absorption and while GSK650394A inhibited SGK1 without affecting PKB, Akti-1/2 inactivated both kinases. CONCLUSION AND IMPLICATIONS While studies undertaken using PI103 and GDC-0941 show that hormone-deprived cells can absorb Na+ independently of PI3K, PI3K seems to be essential for insulin induced Na+ transport. Akti-1/2 does not act as a selective inhibitor of PKB and data obtained using this compound must therefore be treated with caution. GSK650394A, on the other hand, selectively inhibits SGK1 and the finding that GSK650394A suppressed insulin-induced Na+ absorption suggests that this response is dependent upon signalling via PI3K/SGK1. [source]

EFFECT OF BUTYRIC ACID SUPPLEMENTATION ON SERUM AND RENAL ANTIOXIDANT ENZYME ACTIVITIES IN STREPTOZOTOCIN-INDUCED DIABETIC RATS

JOURNAL OF FOOD BIOCHEMISTRY, Issue 2010
A. PUNEETH KUMAR
ABSTRACT Reactive oxygen metabolites, which are constant products of normal aerobic cell metabolism, play a key role in worsening the pathophysiological complications of diabetes. The present investigation was aimed at understanding the effect of butyric acid supplementation along with wheatbran and guar gum on serum and renal antioxidant enzyme activities and lipid peroxidation in streptozotocin (STZ)-induced diabetic rats. Activities of superoxide dismutase, catalase, glutathione peroxidase were evaluated in serum and kidney of control and experimental rats. Results clearly showed that the altered activity of the enzymes during diabetes was significantly ameliorated by butyric acid (500 mg/kg body weight/day) supplementation compared with other experimental groups. Further, the increased lipid peroxidation in serum and kidney of diabetic rats was also significantly reduced in butyric acid-supplemented diabetic rats. The study led us to conclude that butyric acid exert antioxidant property, thereby minimizing oxidative stress induced diabetes and its related complications. PRACTICAL APPLICATIONS Butyric acid , a product of dietary fiber fermentation , is a four-carbon fatty acid, which has wide range of application in disease management. This product is involved in various physiological functions of body like cell differentiation, apoptosis, colonic homeostasis, histone acetylation, etc. It is also known to decrease the incidence of bowel cancer and some of its analogues are shown to selectively improve glucose-stimulated insulin release and glucose tolerance in both normal and diabetic rats. This study aims to evaluate the beneficial effects of butyric acid supplementation on oxidative stress-induced diabetic complications in rats. [source]

Serum and 24-hour Urine Analysis in Adult Cyanotic and Noncyanotic Congenital Heart Disease Patients

CONGENITAL HEART DISEASE, Issue 3 2009
Efrén Martínez-Quintana MD
ABSTRACT Introduction., Glomerulopathy is a complication of congenital heart disease patients. The risk of developing renal impairment is particularly high in cyanotic patients. Objective., The aim of this study was to determine the prevalence of renal dysfunction and microalbumiuria in adult cyanotic and non cyanotic congenital heart disease patients. Methods., Fourteen cyanotic and 22 noncyanotic congenital heart disease patients were studied in the Adult Congenital Heart Disease Unit at the Complejo Hospitalario Universitario Insular-Materno Infantil. Demographic characteristics, complete blood count, and 24-hour urianalysis were obtained, including abdominal ultrasound in those with cyanosis. Results., No differences were seen between age (years) (27.4 ± 8.2; 26.4 ± 8.3; P = .71), sex, size, weight, or glomerular filtration rate (mL/min/1.73 m2) (81.1 ± 22.9 vs. 84.9 ± 9.2, P = .482) between cyanotic and noncyanotic patients. However, Eisenmenger patients had significantly impaired renal function when compared with noncyanotic patients (73.0 ± 17.3 vs. 84.9 ± 9.2 mL/min/1.73 m2, P = .023). Significant differences were obtained in oxygen saturation (%) (83.8 ± 5.8 vs. 97.8 ± 0.8; P = .000), hematocrit (%) (59.3 ± 8.1 vs. 40.9 ± 8.5; P = .000), platelets (103/µL) (161.5 ± 70.5 vs. 277.9 ± 57.6; P = .000), serum uric acid (mg/dL) (7.5 ± 2.3 vs. 5.6 ± 1.5; P = .008) and microalbuminuria (mg/24 hours) (12.8 [0, 700.2] vs. 2.4 [0, 18.9]; P = .000) between cyanotic and noncyanotic patients. Five cyanotic patients (35.7%) had microalbuminuria (>30 mg/24 hours) and three of them (21.4%) proteinuria (>1 g/24 hours). No significant differences were seen between serum and urine parameters between cyanotic patients who had microalbuminuria (>30 mg/24 hours) and those cyanotic patients who did not have it (<30 mg/24 hours). Conclusions., Renal impairment is frequently seen in congenital heart disease patients, being associated occasionally with proteinuria and microalbuminuria in cyanotic ones. [source]

Serum ,-glutamyltransferase within its normal concentration range is related to the presence of diabetes and cardiovascular risk factors

DIABETIC MEDICINE, Issue 9 2005
D.-J. Kim
Abstract Aims Although many studies have reported an association between serum ,-glutamyltransferase (GGT) and cardiovascular risk factors, the mechanism of this relationship has not been clarified. Methods The medical records of 29 959 subjects (age, median 48, range 14,90 years; 16 706 men, 13 253 women) who visited the Center for Health Promotion at Samsung Medical Center for a medical check-up between January 2001 and December 2003, were investigated. Subjects with hepatic enzyme/GGT concentrations higher than three times the upper limit of the reference range, a positive test for hepatitis C virus antibody, a positive test for hepatitis B virus surface antigen, currently taking anti-diabetic/anti-hypertensive/anti-lipid medication, or a white blood cell (WBC) count higher than 10 000 cells/ml, were excluded. The subjects of each gender were classified into five groups according to their serum GGT concentrations, into quartiles of the normal range of GGT (groups 1, 2, 3 and 4) and into a group with elevated GGT (group 5). Results As the group number increased (group 1 , 5), the frequencies of all of the following increased: (i) diabetes and impaired fasting glucose (IFG); (ii) hypertension, obesity (body mass index , 27 kg/m2), dyslipidaemia (LDL-cholesterol , 4.1 mmol/l and/or triglyceride , 2.46 mmol/l, or HDL-cholesterol < 1.16 mmol/l); (iii) metabolic syndrome. Moreover, these significant relationships between GGT concentrations within its normal range and the presence of diabetes/IFG, hypertension, obesity, dyslipidaemia, and metabolic syndrome persisted after adjusting for several clinical and biochemical variables and for the presence of fatty liver based on ultrasonographic findings. Odds ratios (95% CI) for group 4 (highest quartile of normal range of GGT) vs. group 1 (lowest quartile of normal range of GGT); the referent group, were 3.16 (2.15,4.65) for diabetes, 2.24 (1.73,2.90) for IFG, 1.93 (1.59,2.33) for obesity, 1.38 (1.23,1.55) for dyslipidaemia and 2.88 (2.28,3.65) for metabolic syndrome in men. In women, the odds ratios were 2.72 (1.34,5.52), 3.67 (2.26,5.97), 2.10 (1.61,2.74), 1.80 (1.58,2.04) and 3.57 (2.52,5.07), respectively. Conclusions Our data show that, even within its normal range, serum GGT concentrations are closely associated with the presence of diabetes and cardiovascular risk factors, and that these associations are independent of a fatty liver by ultrasonography. [source]

Laparoscopic findings of liver cirrhosis due to non-alcoholic steatohepatitis

DIGESTIVE ENDOSCOPY, Issue 4 2003
Teruki Miyake
A 42-year-old Japanese man was admitted to our hospital for investigation of abnormal liver function tests. He had no history of drug use, and drank little alcohol. Body mass index was 30. Serum was negative for viral markers and autoantibodies. Laparoscopy revealed diffuse small nodules on the liver surface. Liver biopsy revealed small nodules with pericellular fibrosis and macrovesicular fat deposition throughout the acini. Some inflammatory changes were observed. Liver cirrhosis due to non-alcoholic steatohepatitis (NASH) was diagnosed. NASH displays similar histological and laparoscopic characteristics to alcoholic liver diseases. [source]

Detection of C Reactive Protein (CRP) in Serum by an Electrochemical Aptamer-Based Sandwich Assay

ELECTROANALYSIS, Issue 11 2009
Sonia Centi
Abstract A disposable electrochemical assay involving magnetic particles and carbon-based screen-printed electrodes (SPCEs) was developed for the detection of C Reactive Protein (CRP). CRP is a plasma protein and is among the most expressed proteins in acute phase inflammation cases, being a known biomarker for inflammatory states. The assay was based on a sandwich format in which a RNA aptamer was coupled to a monoclonal antibody and alkaline phosphatase (AP) was used as enzymatic label. After the sandwich assay, the modified magnetic beads were captured by a magnet on the surface of a graphite working electrode and the electrochemical detection was thus achieved through the addition of the AP substrate (,-naphthyl-phosphate) and ,-naphthol produced during the enzymatic reaction was detected using differential pulse voltammetry (DPV). The parameters influencing the different steps of the assay were optimized in order to reach the best sensitivity and specificity. With the optimized conditions, the assay was applied to the analysis of CRP free serum and serum samples. [source]

Electrochemical Investigation of Strontium,Metallothionein Interactions , Analysis of Serum and Urine of Patients with Osteoporosis

ELECTROANALYSIS, Issue 3-5 2009
Ivo Fabrik
Abstract The main aim of this paper is to study interaction between strontium and metallothionein (MT), and to determine changes in strontium and thiols (MT, reduced glutathione, cysteine, and homocysteine) level in plasma, serum, and urine samples of patients treated with strontium ranelate (SrR). To investigate the interactions between MT and strontium(II) ions, adsorptive transfer stripping technique coupled with differential pulse voltammetry (DPV) the Brdicka reaction was employed. Besides standard Brdicka signals (Co, RS2Co, Cat1, Cat2, Cat3), we observed new signal related to Sr-MT interaction. Further we investigated the effect of various time of interaction, concentration of strontium(II) ions and temperature of supporting electrolyte on Brdicka signals. Optimal time of interaction was 240,s. Under temperature of supporting electrolyte 20,°C, we measured linear dependence of Cat3 signal height on strontium(II) ions concentration. After that we have investigated the possibility of strontium-MT interactions, we were interested in strontium, MT and low molecular mass thiols levels in serum and urine of patients treated with strontium(II) ions to cure osteoporosis. Strontium concentration determined by atomic absorption spectrometry was 55±5,,g/L before and 10,500±1,400,,g/L at the 30th day of SrR administration. Levels of metallothionein in serum ranged from 0.1 to 6.4,,M. Correlation between serum strontium concentration and MT level was determined and correlation coefficient was R=0.93. [source]

Electrochemical Evaluation of Nucleoside Analogue Lamivudine in Pharmaceutical Dosage Forms and Human Serum

ELECTROANALYSIS, Issue 20 2005
Burcu Dogan
Abstract Lamivudine (LAM) is a synthetic nucleoside analogue with activity against human immunodeficiency virus-type 1 (HIV-1) and Hepatitis B virus (HBV). The aim of this study was to determine LAM levels in serum and pharmaceutical formulations, by means of electrochemical methods using hanging mercury drop electrode (HMDE). On this electrode, LAM undergoes irreversible reduction at the peak potential near Ep,1.26,V (vs. Ag/AgCl/3,M KCl). Reduction LAM signals were measured by cyclic voltammetry (CV), differential pulse voltammetry (DPV) and square-wave voltammetry (OSW). DPV and OSW techniques for the determination of LAM in acetate buffer at pH,4.5, which allows quantitation over the 4×10,6 to 1×10,4,M range in supporting electrolyte for both methods, were proposed. The linear response was obtained in acetate buffer in the ranges of 2×10,6 to 2×10,4,M for spiked serum samples at pH,4.5 for both techniques. The repeatability and reproducibility of the methods for all media were determined. The standard addition method was used in serum. Precision and accuracy were also checked in all media. No electroactive interferences from the endogenous substances were found in serum. With respect to side effects of high doses and short half-life of LAM, a fast and simple detection method is described in this study. [source]

Bone mineral density in familial amyloid polyneuropathy and in other neuromuscular disorders

EUROPEAN JOURNAL OF NEUROLOGY, Issue 6 2005
I. M. Conceição
Neuromuscular diseases are a known risk factor for immobilization-induced osteoporosis. The aim of the study was to analyse bone mineral density (BMD) in patients with familial amyloid polyneuropathy (FAP) type I (Val30 Met) and to compare them with a population of patients with other neuromuscular disorders. We studied 24, ambulatory, neuromuscular patients, all men and premenopausal women. We included 12 FAP patients (GI) and 12 patients with other disorders (GII). Clinical data included age, sex, height, weight, alcohol intake, smoking, calcium intake, physical activity and history of fractures. Serum and urinary calcium, osteocalcin, bone alkaline phosphatase, parathyroid hormone, thyroid stimulating hormone and urinary N-telopeptide cross-linked type 1 collagen were determined in all patients. Bone mineral density of lumbar spine, hip and wrist were determined by dual energy X-ray absorptiometry scan. No statistical differences were found in clinical or analytic data between the two groups, except for body mass index and calciuria, which were lower in GI. In GI, 54.5% were osteoporotic, against 23.1% in GII (P = 0.04). Bone mineral density was lower in GI when compared with GII, and tended to decrease with disease duration. Decreased BMI and the early autonomic involvement in GI probably explain the results. The prevention and early treatment of osteoporosis, in FAP patients should be considered a priority. [source]

Serum or target deprivation-induced neuronal death causes oxidative neuronal accumulation of Zn2+ and loss of NAD+

EUROPEAN JOURNAL OF NEUROSCIENCE, Issue 6 2010
Christian T. Sheline
Abstract Trophic deprivation-mediated neuronal death is important during development, after acute brain or nerve trauma, and in neurodegeneration. Serum deprivation (SD) approximates trophic deprivation in vitro, and an in vivo model is provided by neuronal death in the mouse dorsal lateral geniculate nucleus (LGNd) after ablation of the visual cortex (VCA). Oxidant-induced intracellular Zn2+ release ([Zn2+]i) from metallothionein-3 (MT-III), mitochondria or ,protein Zn2+', was implicated in trophic deprivation neurotoxicity. We have previously shown that neurotoxicity of extracellular Zn2+ required entry, increased [Zn2+]i, and reduction of NAD+ and ATP levels causing inhibition of glycolysis and cellular metabolism. Exogenous NAD+ and sirtuin inhibition attenuated Zn2+ neurotoxicity. Here we show that: (1) Zn2+ is released intracellularly after oxidant and SD injuries, and that sensitivity to these injuries is proportional to neuronal Zn2+ content; (2) NAD+ loss is involved , restoration of NAD+ using exogenous NAD+, pyruvate or nicotinamide attenuated these injuries, and potentiation of NAD+ loss potentiated injury; (3) neurons from genetically modified mouse strains which reduce intracellular Zn2+ content (MT-III knockout), reduce NAD+ catabolism (PARP-1 knockout) or increase expression of an NAD+ synthetic enzyme (Wlds) each had attenuated SD and oxidant neurotoxicities; (4) sirtuin inhibitors attenuated and sirtuin activators potentiated these neurotoxicities; (5) visual cortex ablation (VCA) induces Zn2+ staining and death only in ipsilateral LGNd neurons, and a 1 mg/kg Zn2+ diet attenuated injury; and finally (6) NAD+ synthesis and levels are involved given that LGNd neuronal death after VCA was dramatically reduced in Wlds animals, and by intraperitoneal pyr vate or nicotinamide. Zn2+ toxicity is involved in serum and trophic deprivation-induced neuronal death. [source]

A novel N-terminal hydrophobic motif mediates constitutive degradation of serum- and glucocorticoid-induced kinase-1 by the ubiquitin,proteasome pathway

FEBS JOURNAL, Issue 13 2006
Agata M. Bogusz
Serum- and glucocorticoid-induced protein kinase-1 (SGK-1) plays a critical role in regulation of the epithelial sodium channel, ENaC. SGK-1 also shares significant catalytic domain homology with protein kinase B (PKB/AKT-1) and is a downstream effector of antiapoptotic phosphoinositide 3-kinase signaling. Steady-state levels of an active SGK-1 are tightly regulated by rapid transcriptional activation and post-translational modification including phosphorylation. We show here that endogenous SGK-1 protein is polyubiquitinated and rapidly degraded by the 26S proteasome. In contrast to other rapidly degraded kinases, neither the catalytic activity of SGK-1 nor activation site phosphorylation was required for its ubiquitin modification and degradation. Instead, SGK-1 degradation required a lysine-less six-amino-acid (amino acids 19,24) hydrophobic motif (GMVAIL) within the N-terminal domain. Deletion of amino acids 19,24 significantly increased the half-life of SGK1 and prevented its ubiquitin modification. Interestingly, this minimal region was also required for the association of SGK-1 with the endoplasmic reticulum. Ubiquitin modification and degradation of SGK-1 were increasingly inhibited by the progressive mutation of six N-terminal lysine residues surrounding the GMVAIL motif. Mutation of all six lysines to arginine did not disrupt the subcellular localization of SGK-1 despite a significant decrease in ubiquitination, implying that this modification per se was not required for targeting to the endoplasmic reticulum. These results suggest that constitutive ubiquitin-mediated degradation of SGK-1 is an important mechanism regulating its biological activity. [source]

DNA Detection: Intercalating Dye Harnessed Cationic Conjugated Polymer for Real-Time Naked-Eye Recognition of Double-Stranded DNA in Serum (Adv. Funct.

ADVANCED FUNCTIONAL MATERIALS, Issue 9 2009
Mater.
On page 1371, Liu Bin and Pu Kan-Yi demonstrate multicolor detection of double-stranded DNA in biological media using an intercalating-dye-harnessed cationic conjugated polymer. As demonstrated in the cover image, the intercalating-dye-harnessed polymer emits blue fluorescence both in the absence and presence of single-stranded DNA in serum-containing solution, while its fluorescence gradually turns from blue to dark yellow with increasing double-stranded DNA concentration. [source]

Intercalating Dye Harnessed Cationic Conjugated Polymer for Real-Time Naked-Eye Recognition of Double-Stranded DNA in Serum

ADVANCED FUNCTIONAL MATERIALS, Issue 9 2009
Kan-Yi Pu
Abstract Thiazole orange (TO), an intercalating dye, is integrated into cationic poly(fluorene- alt -phenylene) (PFP) to develop a macromolecular multicolor probe (PFPTO) for double-stranded DNA (dsDNA) detection. This polymer design not only takes advantage of the high affinity between TO and dsDNA to realize dsDNA recognition in biological media, but also brings into play the light-harvesting feature of conjugated polymers to amplify the signal output of TO in situ. PFPTO differentiates dsDNA from single-stranded DNA (ssDNA) more effectively upon excitation of the conjugated backbone relative to that upon direct excitation of TO as a result of efficient fluorescence resonance energy transfer from the polymer backbone to the intercalated TO. In the presence of dsDNA, energy transfer within PFPTO is more efficient as compared to that for free TO/PFP system, which leads to better dsDNA discriminability for PFPTO in contrast to that for TO/PFP. The distinguishable fluorescent color for PFPTO solutions in the presence of dsDNA allows naked-eye detection of dsDNA with the assistance of a hand-held UV lamp. The significant advantage of this macromolecular fluorescent probe is that naked-eye detection of label-free dsDNA can be performed in biological media in real-time. [source]

Serum and forskolin cooperate to promote G1 progression in Schwann cells by differentially regulating cyclin D1, cyclin E1, and p27Kip expression

GLIA, Issue 16 2007
Jared Iacovelli
Abstract Proliferation of Schwann cells in vitro, unlike most mammalian cells, is not induced by serum alone but additionally requires cAMP elevation and mitogenic stimulation. How these agents cooperate to promote progression through the G1 phase of the cell cycle is unclear. We studied the integrative effects of these compounds on receptor-mediated signaling pathways and regulators of G1 progression. We show that serum alone induces strong cyclical expression of cyclin D1 and E1, 6 and 12 h after addition, respectively. Serum also promotes strong but transient erbB2, ERK, and Akt phosphorylation, but Schwann cells remain arrested in G1 due to high levels of the inhibitor, p27Kip. Forskolin with serum promotes G1 progression in 22% of Schwann cells between 18 and 24 h by inducing a steady decline in p27Kip levels that reaches a nadir at 12 h coinciding with peak cyclin E1 expression. Forskolin also delays neuregulin-induced loss of erbB2 receptors allowing strong acute activation of PI3K, sustained erbB2 phosphorylation and G1 progression in 31% of Schwann cells. We find that the ability of forskolin to decrease p27Kip is associated with its ability to decrease Krox-20 expression that is induced by serum and further increased by neuregulin. Our results explain why serum is required but insufficient to stimulate proliferation and identify two routes by which forskolin promotes proliferation in the presence of serum and neuregulin. These findings provide insights into how G1 progression and, cell cycle arrest leading to myelination are regulated in Schwann cells. © 2007 Wiley-Liss, Inc. [source]

Prognostic significance of serum p53 protein and p53 antibody in patients with surgical treatment for head and neck squamous cell carcinoma

HEAD & NECK: JOURNAL FOR THE SCIENCES & SPECIALTIES OF THE HEAD AND NECK, Issue 4 2001
Vivian Chow M. Phil
Abstract Objectives This study aims at investigating the prognostic values of serum p53 protein and anti-p53 antibody in patients undergoing surgical treatment for head and neck squamous cell carcinoma (HNSCC). Methods Serum p53 protein and anti-p53 antibody concentrations were determined by an enzyme-linked immunosorbent assay (ELISA) in 75 patients with HNSCC and 28 healthy controls. In 28 patients, formalin-fixed tumor tissues were also available for immunohistochemical staining by an anti-p53 DO7 monoclonal antibody. The results were correlated with the clinicopathologic parameters. Results The mean preoperative serum concentration of p53 protein in patients with HNSCC was significantly higher than healthy controls (59.45 pg/mL vs 16.4 pg/mL, p = .007). Preoperative serum p53 antibody was present in 23 (31%) patients and was present in one healthy control. Eighteen (62%) tumor tissues showed p53 overexpression by immunohistochemistry. The presence of serum anti-p53 antibody before operation was associated with a significantly higher incidence (65%) of nodal metastasis compared with 27% nodal metastasis in patients with absence of serum anti-p53 antibody (p = .002). Conclusion Preoperative serum p53 antibody was a significant prognostic factor for nodal metastasis of HNSCC. © 2001 John Wiley & Sons, Inc. Head Neck 23: 286,291, 2001. [source]

Role of Immune Serum in the Killing of Helicobacter pylori by Macrophages

HELICOBACTER, Issue 3 2010
Stacey Keep
Abstract Background:,Helicobacter pylori infection can lead to the development of gastritis, peptic ulcers and gastric cancer, which makes this bacterium an important concern for human health. Despite evoking a strong immune response in the host, H. pylori persists, requiring complex antibiotic therapy for eradication. Here we have studied the impact of a patient's immune serum on H. pylori in relation to macrophage uptake, phagosome maturation, and bacterial killing. Materials and Methods:, Primary human macrophages were infected in vitro with both immune serum-treated and control H. pylori. The ability of primary human macrophages to kill H. pylori was characterized at various time points after infection. H. pylori phagosome maturation was analyzed by confocal immune fluorescence microscopy using markers specific for H. pylori, early endosomes (EEA1), late endosomes (CD63) and lysosomes (LAMP-1). Results:, Immune serum enhanced H. pylori uptake into macrophages when compared to control bacteria. However, a sufficient inoculum remained for recovery of viable H. pylori from macrophages, at 8 hours after infection, for both the serum-treated and control groups. Both serum-treated and control H. pylori phagosomes acquired EEA1 (15 minutes), CD63 and LAMP-1 (30 minutes). These markers were then retained for the rest of an 8 hour time course. Conclusions:, While immune sera appeared to have a slight positive effect on bacterial uptake, both serum-treated and control H. pylori were not eliminated by macrophages. Furthermore, the same disruptions to phagosome maturation were observed for both serum-treated and control H. pylori. We conclude that to eliminate H. pylori, a strategy is required to restore the normal process of phagosome maturation and enable effective macrophage killing of H. pylori, following a host immune response. [source]

Systemic Immunization with Unadjuvanted Whole Helicobacter pylori Protects Mice Against Heterologous Challenge

HELICOBACTER, Issue 6 2008
Stacey N. Harbour
Abstract Background:, Adjuvant-free vaccines have many benefits, including decreased cost and toxicity. We examined the protective effect of systemic vaccination with adjuvant-free formalin-fixed Helicobacter pylori or bacterial lysate and the ability of this vaccine to induce protection against heterologous challenge. Materials and Methods:, Mice were vaccinated subcutaneously with H. pylori 11637 lysate or formalin-fixed bacteria, with or without ISCOMATRIXTM adjuvant, then orally challenged with H. pylori SS1. Serum was taken prior to challenge to examine specific antibody levels induced by the vaccinations, and protection was assessed by colony-forming assay. Results:, Vaccination with H. pylori 11637 lysate or formalin-fixed bacteria delivered systemically induced significantly higher levels of Helicobacter -specific serum IgG than the control, unvaccinated group and orally vaccinated group. After heterologous challenge with H. pylori SS1, all vaccinated groups had significantly lower levels of colonization compared with unvaccinated, control mice, regardless of the addition of adjuvant or route of delivery. Protection induced by systemic vaccination with whole bacterial preparations, without the addition of adjuvants, was only associated with a mild cellular infiltration into the gastric mucosa, with no evidence of atrophy. Conclusions:, Subcutaneous vaccination using unadjuvanted formalin-fixed H. pylori has the potential to be a simple, cost-effective approach to the development of a Helicobacter vaccine. Importantly, this vaccine was able to induce protection against heterologous challenge, a factor that would be crucial in any human Helicobacter vaccine. Further studies are required to determine mechanisms of protection and to improve protective ability. [source]

Serum cytokeratin-18 fragment level: A noninvasive biomarker for not only nonalcoholic steatohepatitis, but also alcoholic steatohepatitis,

HEPATOLOGY, Issue 5 2010
M.D., Xiaohua Li Ph.D.
No abstract is available for this article. [source]

Serum laminin-2 and hyaluronan predict severe liver fibrosis in children with chronic hepatitis B

HEPATOLOGY, Issue 3 2004
Dariusz M. Lebensztejn
First page of article [source]

Early identification of recipients with progressive histologic recurrence of hepatitis C after liver transplantation

HEPATOLOGY, Issue 5 2000
Raghavakaimal Sreekumar
Approximately half of patients undergoing liver transplantation (LT) for hepatitis C virus (HCV) develop histologic evidence of recurrence within the first postoperative year. Early identification of recipients at risk for more severe recurrence of HCV may be useful in selecting patients for antiviral therapy. We determined whether recipients at greatest risk for more severe recurrence of HCV can be identified by pre- and/or early post-LT HCV-RNA levels in serum or tissue. Serum and tissue samples were prospectively collected pre-LT and at 7 days, 4 months, 1 year, and at 3 years posttransplantation from patients undergoing LT for HCV. Hepatitis activity index (HAI) and fibrosis stage (FS) were assessed in all liver biopsies. Forty-seven patients (32 men) were studied. Higher HCV-RNA levels at 4 months post-LT (,109 copies/mL, n = 29) were associated with higher HAI at 1 year and at 3 years post-LT. The HAI seen on protocol biopsies at 4 months correlated significantly with fibrosis stage (FS) at 1 year (r = .56, P , .001) and 3 years (r = .53, P = .002). Higher HCV-RNA levels at 7 days and 4 months post-LT were sensitive (66% and 84%, respectively) and specific (92% and 63%, respectively) in identifying recipients with an HAI greater than 3 at 3 years. Higher pre- and early post-LT HCV-RNA levels are associated with more severe recurrence of HCV. The correlation of early HAI with subsequent FS suggests that higher mean HAI will eventually translate into more advanced stages of fibrosis. Patients at risk for more severe post-LT recurrence of HCV can be identified by early posttransplant HCV-RNA levels. [source]

Local and systemic interleukin-18 and interleukin-18-binding protein in children with inflammatory bowel disease

INFLAMMATORY BOWEL DISEASES, Issue 1 2008
Steven T. Leach
Abstract Background: Interleukin-18 (IL-18) is increased in the inflamed mucosa of patients with Crohn's disease (CD). The balance between this pleiotropic proinflammatory cytokine and its natural inhibitor, IL-18-binding protein (IL-18BP), may contribute to the pathogenesis of inflammatory bowel disease (IBD). Methods: Serum and mucosal biopsies were collected from children with IBD, from children with celiac disease, and from controls. Biopsies were maintained in culture for 24 hours, and supernatant was collected. Serum and supernatant IL-18 and IL-18BPa concentrations were measured by immunoassay. Disease activity score (PCDAI) and standard serum inflammatory markers (albumin, platelets, ESR, and CRP) were recorded. Results: Serum IL-18 was greater in children with CD (537 pg/mL) than in controls (335 pg/mL; P < 0.05) but not in children with ulcerative colitis (UC) or IBD type unclassified (IBDU). Mucosal IL-18 was greater in children with CD and UC/IBDU than in controls (P < 0.01). Serum IL-18BPa was increased in children with CD compared with that in controls (3.9 versus 2.6 ng/mL; P < 0.05), but was not elevated in children with UC/IBDU. Furthermore, calculated free-serum IL-18 was elevated in CD, but not UC/IBDU, compared with that in controls (P = 0.001). Total and free-serum IL-18 were elevated in severe CD relative to in mild/moderate disease. Conclusions: IL-18, produced in the colons of children with IBD, may contribute to local inflammatory changes. Systemic IL-18 level may be a useful indicator of gut inflammation. Furthermore, free IL-18 is greatly elevated in children with CD, suggesting that compensatory increases in IL-18BPa are insufficient. Further exploration of the role of this cytokine in the pathogenesis of IBD is now required. (Inflamm Bowel Dis 2007) [source]

Simple Fabrication of Antibody Microarrays on Nonfouling Polymer Brushes with Femtomolar Sensitivity for Protein Analytes in Serum and Blood

ADVANCED MATERIALS, Issue 19 2009
Angus Hucknall
A multianalyte antibody array that is spotted on a poly(oligo(ethylene glycol) methacrylate) brush 100,nm thick, grown on glass via surface-initiated atom transfer radical polymerization, has femtomolar limit-of-detection (LOD) of cytokines in serum and whole blood, and a dynamic range of six orders of magnitude for a range of protein analytes. [source]

Vitamin D deficiency in a multinational refugee population

INTERNAL MEDICINE JOURNAL, Issue 12 2007
H. D. Wishart
Abstract Background: Populations with increased skin pigmentation who have migrated to countries of high latitude are at increased risk of low vitamin D. This study aimed to determine the prevalence of low vitamin D among the refugee population arriving in New Zealand. Methods: An audit of all refugees arriving at the national refugee resettlement centre from May 2004 to May 2005 was carried out. Serum 25-hydroxyvitamin D3 levels were measured and defined as normal (50,150 nmol/L) or low, with low subdivided into insufficient (25 to <50 nmol/L) and deficient (<25 nmol/L). Whether vitamin D status varied with age and sex was determined. Results: Vitamin D was measured in 869 (99%) of the refugees and was low in 470 (54%, 95% confidence interval (CI) 51,57%). It was insufficient in 323 (37%, 95%CI 34,41%) and deficient in 147 (17%, 95%CI 15,20%). Female sex was associated with at least a 10 times increased risk of vitamin D deficiency (relative ratio 13.93, 95%CI 10.15,17.96). Women aged between 17 and 45 years and men aged 46 years and more were at greatest risk. Conclusion: Poor vitamin D status is prevalent among refugees arriving in New Zealand. Women, particularly those of child-bearing age are at greatest risk. Screening and ongoing surveillance for vitamin D deficiency should be considered for all recent refugee immigrants to New Zealand. [source]

Emergency Department Lactate Is Associated with Mortality in Older Adults Admitted With and Without Infections

ACADEMIC EMERGENCY MEDICINE, Issue 3 2010
Daniel A. Del Portal MD
Abstract Objectives:, Serum lactate values in the emergency department (ED) have been associated with mortality in diverse populations of critically ill patients. This study investigates whether serum lactate values measured in the ED are associated with mortality in older patients admitted to the hospital, both with and without infections. Methods:, This is a retrospective cohort study performed at two urban teaching hospitals. The study population includes 1,655 older ED patients (age , 65 years) over a 3-year period (2004,2006) who had serum lactate measured prior to admission. The presence or absence of infection was determined by review of International Classification of Diseases Ninth Revision (ICD-9) admission diagnosis codes. Mortality during hospitalization was determined by review of inpatient records. Mortality at 30 and at 60 days was determined using a state death registry. Results:, In patients with infections, increasing serum lactate values of ,2.0 mmol/L were linearly associated with relative risk (RR) of mortality during hospitalization (RR = 1.9 to 3.6 with increasing lactate), at 30 days (RR = 1.7 to 2.6), and at 60 days (RR = 1.4 to 2.3) when compared to patients with serum lactate levels of <2.0 mmol/L. In patients without infections, a similar association was observed (RR = 1.1 to 3.9 during hospitalization, RR = 1.2 to 2.6 at 30 days, RR = 1.1 to 2.4 at 60 days). In both groups of patients, serum lactate had a greater magnitude of association with mortality than either of two other commonly ordered laboratory tests, leukocyte count and serum creatinine. Conclusions:, Higher ED lactate values are associated with greater mortality in a broad cohort of admitted patients over age 65 years, regardless of the presence or absence of infection. ACADEMIC EMERGENCY MEDICINE 2010; 17:260,268 © 2010 by the Society for Academic Emergency Medicine [source]

Heterogeneity in Serum 25-Hydroxy-Vitamin D Response to Cholecalciferol in Elderly Women with Secondary Hyperparathyroidism and Vitamin D Deficiency

JOURNAL OF AMERICAN GERIATRICS SOCIETY, Issue 8 2010
Andrea Giusti MD
OBJECTIVES: To compare the effects on parathyroid hormone (PTH) and 25-hydroxy-vitamin D (25(OH)D) of two dosing regimens of cholecalciferol in women with secondary hyperparathyroidism (sHPTH) and hypovitaminosis D and to investigate variables affecting 25(OH)D response to cholecalciferol. DESIGN: Randomized-controlled trial with 6-month follow-up. SETTING: Two osteoporosis centers in northern Italy. PARTICIPANTS: Sixty community-dwelling women aged 65 and older with sHPTH and hypovitaminosis D, creatinine clearance greater than 65 mL/min and without diseases or drugs known to influence bone and vitamin D metabolism. INTERVENTION: Cholecalciferol 300,000 IU every 3 months, once at baseline and once at 3 months (intermittent D3 group) or cholecalciferol 1,000 IU/day (daily D3 group). MEASUREMENTS: Serum PTH, 25(OH)D, calcium, bone-specific alkaline phosphatase, ,-C-terminal telopeptide of type I collagen, phosphate, 24-hour urinary calcium excretion. RESULTS: The two groups had similar baseline characteristics. All participants had vitamin D deficiency [25(OH)D<20 ng/mL)], and 36 subjects (60%) had severe deficiency (<10 ng/mL), with no difference between the groups (severe deficiency: intermittent D3 group, n=18; daily D3 group, n=18). After 3 and 6 months, both groups had a significant increase in 25(OH)D and a reduction in PTH. Mean absolute increase±standard deviation of 25(OH)D at 6 months was higher in the intermittent D3 group (22.7±11.8 ng/mL) than in the daily D3 group (13.7±6.7 ng/mL, P<.001), with a higher proportion of participants in the intermittent D3 group reaching desirable serum concentration of 25(OH)D , 30 ng/mL (55% in the intermittent D3 group vs 20% in the daily D3 group, P<.001). Mean percentage decrease of PTH in the two groups was comparable, and at 6 months, a similar proportion of participants reached normal PTH values. 25(OH)D response to cholecalciferol showed a wide variability. In a logistic regression analysis, body mass index and type of treatment appeared to be significantly associated with normalization of 25(OH)D values. CONCLUSION: Cholecalciferol 300,000 IU every 3 months was more effective than 1,000 IU daily in correcting vitamin D deficiency, although the two groups achieved similar effects on PTH at 6 months. Only 55% of the higher-dose intermittent group reached desirable concentrations of 25(OH)D, suggesting that yet-higher doses will be required for adequate vitamin D repletion. [source]

Effect of Fracture on Bone Turnover Markers: A Longitudinal Study Comparing Marker Levels Before and After Injury in 113 Elderly Women,

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 8 2007
Kaisa K Ivaska PhD
Abstract In this longitudinal, prospective, and population-based study (n = 1044), seven BTMs were assessed before and after trauma in 113 elderly women (85 with fractures). Markers were not altered in the immediate postfracture period but were clearly elevated during fracture repair. Recent fracture should thus be taken into account when markers are used in clinical practice. Introduction: Fracture may influence the levels of bone turnover markers (BTM) and have implications for their use in clinical practice. In this longitudinal, prospective, and population-based study, we assessed prefracture levels of BTMs and compared them with postfracture levels of the same individuals immediately after fracture and during fracture repair. This is the first study in which the effect of fracture on bone markers has been evaluated with prefracture samples available. Materials and Methods: Serum and urine were collected at the emergency unit from 85 women (77.9 ± 1.8 yr) who sustained a fracture after low-energy trauma and 28 controls (77.8 ± 2.0 yr) with similar trauma but no fracture. All were participants of the Malmö OPRA study (n = 1044), and pretrauma samples were collected 1.05 ± 0.85 yr before. Bone turnover was assessed by seven different BTMs reflecting different stages of bone metabolism {C-terminal cross-linked telopeptides of type I collagen [S-CTX], S-TRACP5b, N-terminal propeptides of type I collagen [S-PINP], serum osteocalcin (S-OC[1,49] and S-TotalOC), urinary deoxypyridinoline [U-DPD], and urinary osteocalcin [U-OC]}. Results: BTMs sampled within a few hours after fracture were not altered from preinjury levels. Both bone formation and bone resorption markers were, however, significantly increased 4 mo after fracture. The elevation was most pronounced after hip fracture. Bone turnover remained elevated up to 12 mo after fracture. Conclusions: We believe this study extends our knowledge on the skeletal postfracture metabolic processes. In addition, it may provide a basis for future means to monitor pharmacological intervention promoting fracture healing. [source]

Cross-Sectional Evaluation of Bone Metabolism in Men,

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 9 2001
P. Szulc
Abstract There are relatively few data concerning age-related changes of bone turnover in men. The aim of the study was to evaluate age-related changes of the levels of serum and urinary biochemical markers of bone metabolism in a large cohort of 934 men aged 19,85 years and to investigate their association with bone mineral density (BMD). Bone formation was evaluated using serum levels of osteocalcin (OC), bone alkaline phosphatase (BAP), and N-terminal extension propeptide of type I collagen (PINP). Bone resorption was evaluated by measurement of urinary excretion of ,-isomerized C-terminal telopeptide of collagen type I (,-CTX) of free deoxypyridinoline (fDpyr) and total Dpyr (tDPyr) and of the serum level of ,-CTX. Levels of biochemical bone markers were very high in young men and decreased rapidly until the age of 40 years and then more slowly until 60 years of age. After the age of 60 years, markers of bone formation remained stable while resorption markers showed a moderate and variable increase with aging. Serum and urinary ,-CTX levels were elevated only in about 5% of elderly men. The age-related increase of urinary excretion of tDpyr and of its free and peptide-bound fractions was related to the presence of elevated levels in a subgroup of about 15% of elderly men. Before 60 years of age, levels of biochemical bone markers were not correlated with BMD, whereas after 60 years of age, they were correlated negatively with BMD. After adjustment for age and body weight, BMD in men with the highest levels of biochemical bone markers (i.e., in the upper quartile) was 1.8,12.5% (i.e., 0.25,0.89 SD) lower than in men with levels of biochemical bone markers in the lowest quartile. In conclusion, bone turnover in men is high in young adults and decreases to reach a nadir at 55,60 years of age. After the age of 60 years, bone resorption markers,but not bone formation markers,increase in some men and are associated with lower BMD, suggesting that this imbalance is responsible for increasing bone loss in elderly men. [source]

Serum and glucocorticoid-regulated protein kinases: Variations on a theme

JOURNAL OF CELLULAR BIOCHEMISTRY, Issue 6 2006
Maude Tessier
Abstract The phosphatidylinositol 3, kinase (PI3K)-signaling pathway plays a critical role in a variety of cellular responses such as modulation of cell survival, glucose homeostasis, cell division, and cell growth. PI3K generates important lipid second messengers,phosphatidylinositides that are phosphorylated at the 3, position of their inositol ring head-group. These membrane restricted lipids act by binding with high affinity to specific protein domains such as the pleckstrin homology (PH) domain. Effectors of PI3K include molecules that harbor such domains such as phosphoinositide-dependent kinase (PDK1) and protein kinase B (PKB), also termed Akt. The mammalian genome encodes three different PKB genes (,, ,, and ,; Akt1, 2, and 3, respectively) and each is an attractive target for therapeutic intervention in diseases such as glioblastoma and breast cancer. A second family of three protein kinases, termed serum and glucocorticoid-regulated protein kinases (SGKs), is structurally related to the PKB family including regulation by PI3K but lack a PH domain. However, in addition to PH domains, a second class of 3, phosphorylated inositol phospholipid-binding domains exists that is termed Phox homology (PX) domain: this domain is found in one of the SGKs (SGK3). Here, we summarize knowledge of the three SGK isoforms and compare and contrast them to PKB with respect to their possible importance in cellular regulation and potential as therapeutic targets. J. Cell. Biochem. © 2006 Wiley-Liss, Inc. [source]

Suppressive role of endogenous regucalcin in the enhancement of protein kinase activity with proliferation of cloned rat hepatoma cells (H4-II-E)

JOURNAL OF CELLULAR BIOCHEMISTRY, Issue S36 2001
Shyuichiroh Inagaki
Abstract The role of endogenous regucalcin, which is a regulatory protein in calcium signaling, in the regulation of protein kinase activity in the proliferation of the cloned rat hepatoma cells (H4-II-E) was investigated. Hepatoma cells were cultured for 6,72,h in the presence of fetal bovine serum (FBS; 1 or 10%). The number of cells and protein kinase activity in the 5500,g supernatant of cell homogenate was significantly increased 24 and 48,h after the culture with FBS (1 or 10%); the culture with 10% FBS was potent effect as compared with that of 1% FBS. FBS (10%)-increased protein kinase activity preceded a significant elevation of cell number of 6,h after culture. Serum stimulation-induced increase in protein kinase activity was significantly decreased in the presence of trifluoperazine (50,,M), staurosporine (10,6,M) or genistein (10,5,M) in the enzyme reaction mixture. The presence of anti-regucalcin monoclonal antibody (40 or 80,ng/ml) in the reaction mixture caused a significant increase in protein kinase activity in the cells cultured with FBS (1 or 10%). This increase was completely blocked by addition of regucalcin (10,6,M), which can reveal an inhibitory effect on protein kinase activity. Moreover, the effect of antibody in increasing protein kinase activity was significantly inhibited in the presence of trifluoperazine, staurosporine, or genistein, indicating that endogenous regucalcin has an inhibitory effect on Ca2+/calmodulin-dependent protein kinase, protein kinase C, and protein tyrosine kinase. The present study suggests that endogenous regucalcin plays a suppressive role in the enhancement of various protein kinase activities associated with a proliferation of the cloned rat hepatoma cells (H4-II-E). J. Cell. Biochem. 36: 12-18, 2001. © 2001 Wiley-Liss. Inc. [source]

Factors affecting the serum gastrin 17 level: an evidence-based analysis of 3906 serum samples among Chinese

JOURNAL OF DIGESTIVE DISEASES, Issue 2 2007
Zhong ZHANG
OBJECTIVE: To investigate the influence of gender, age, site of lesion, disease type and Helicobacter pylori (H. pylori) infection on the human serum gastrin-17 level and to study the diagnostic value of serum gastrin-17 in gastric precancerous lesions and gastric cancer. METHODS: Serum gastrin-17 and serum H. pylori IgG antibody were detected by the ELISA method. The different gastric disease groups were confirmed by endoscopy and histopathology. RESULTS: Among the 3906 serum samples according to the gender, age, site of lesion and the data of different gastric disease groups, the serum gastrin-17 level was markedly higher in people ,60 years old than that in younger age groups. The serum gastrin-17 level increased progressively in the following order: healthy control group, nonatrophic gastritis group, gastric ulcer group, and the serum gastrin-17 level was higher in the atrophic gastritis with dysplasia group than that without it, the lowest level being in the gastric cancer group. Among the 2946 serum samples matched with the site of the lesion, the serum gastrin-17 level was higher in those with antral diseases than in those with gastric corpus diseases. Among the 3805 serum samples matched with the H. pylori infection data, the serum gastrin-17 level was higher in the H. pylori -positive group than in the H. pylori -negative group. CONCLUSIONS: In people over 60 years of age, the serum gastrin-17 level tends to increase. In subjects with precancerous gastric lesions, it may increase significantly with the progression of gastric disease, and ultimately decrease in gastric cancer. Serum gastrin-17 is a good biomarker to differentiate benign from malignant gastric diseases. The site of the gastric lesions is an important factor affecting the serum gastrin-17 level, whereas H. pylori infection is usually associated with its increment. [source]