L-Carnitine Supplementation Increases Trimethylamine-N-Oxide but not Markers of Atherosclerosis in Healthy Aged Women.

BACKGROUND: L-carnitine can be metabolized to trimethylamine N-oxide (TMAO), a molecule that promotes atherogenesis through its interaction with macrophages and lipid metabolism. OBJECTIVE: The aim of the present study was to assess whether L-carnitine supplementation may promote changes in selected serum biomarkers of atherosclerosis. METHODS: Before the start, in the mid-point and after completing the 24-weeks supplementation protocol, fasting blood samples were taken from the antecubital vein. Plasma free L-carnitine and TMAO were determined by the UPLC/MS/MS method. Serum proteins were determined by the enzyme immunoassay method using commercially available kits. Total cholesterol, high-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol, and triglycerides have been determined using standard automatic analyzer. RESULTS: L-carnitine supplementation elevated fasting plasma carnitine in the mid-point of our study and it remained increased until the end of supplementation period. Moreover, it induced tenfold increase in plasma TMAO concentration but did not affect serum C-reactive protein, interleukin-6, tumour necrosis factor-α, L-selectin, P-selectin, vascular cell adhesion molecule-1, intercellular adhesion molecule-1 or lipid profile markers. CONCLUSION: We demonstrated that -although oral L-carnitine supplementation significantly -increased plasma TMAO concentration, no lipid profile changes or other markers of adverse cardiovascular events were detected in healthy aged women over the period of 24 weeks.

Obesity and diet awareness among Polish children and adolescents in small towns and villages.

BACKGROUND: In addition to genetic predispositions and environmental factors, healthy lifestyle education is very important for children and adolescents. The purpose of this research was to estimate the number of overweight and obese children and adolescents from small towns and villages and to find out an association between health awareness in children and the risk of becoming overweight or obese. METHODS: The research was conducted in 1,515 healthy children aged 6-18 years from small towns and villages in Poland. Overweight was diagnosed when BMI for age and sex was over the 90th percentile; obesity--when it was over the 95th percentile. The study consisted of a lifestyle interview and anthropometrical measurements. The lifestyle interview was conducted with the use of an anonymous questionnaire form and included questions about food frequency, diet habits and physical activity. The research was analysed using the SAS System for Windows, release 8.02. RESULTS: Overweight status was diagnosed in 9.0% and obesity in 5.1% of respondents. Excess body mass was statistically more frequently diagnosed in girls than in boys aged 14-18 years. Girls of this age group significantly more frequently chose wholemeal bread, smoked sausages, meat and poultry as products that are believed to keep them fit. Older children substantially more often indicated that stress, smoking cigarettes, consuming fatty meat, sweets, being obese, and a lack of physical activity are factors that damage health. Boys spent more time in front of a computer or TV than girls; in the older group of children, the phenomenon even intensified. CONCLUSION: Awareness of healthy lifestyle behaviour is not sufficient to maintain optimal body mass. Knowledge about proper eating habits is better among girls than among boys, especially in the older age groups. However, in older groups, there was less physical activity due to spending more time in front of TV or the computer. High percentage of obese/overweight children and insufficient knowledge of nutrition may consequently result in increased risk of cardio-vascular diseases in adult population.

The concentration of 12-lipoxygenase in platelet rich plasma as an indication of cancer of the prostate.

AIM OF THE STUDY: The aim of this study was to determine whether measuring concentrations of 12-LOX in platelet-rich plasma patients can:Differentiate between the group of patients with prostate cancer and healthy men.Correlate the degree of severity of the disease and the concentration of the enzyme. MATERIAL AND METHODS: The study group comprised 88 men (40-88 years), including 54 patients diagnosed with prostate cancer. The population was divided into 4 groups:group 1 (22 men, aged 55-84 years) -with a negative biopsy,group 2 (36 men, aged 54-88 years) - with a positive biopsy result,group 3 (18 participants aged 58-83) - patients with cancer metastatic disease,group 4 of healthy men (12 people aged 40-66 years) - biopsy was not performed. Routine PSA, morphology and CRP analysis were performed and platelet rich plasma was used for 12(S)LOX determination using an ELISA kit. RESULTS: There was a weak (r = 0.0487) positive correlation between the number of blood platelets and plasma 12(S)LOX.An inverse relationship between 12(S)LOX and Gleason grade was found.Heterogeneity of 12(S)LOX in the group with prostate cancer metastatic disease may suggest differences in the response to the treatment carried out.There were no statistically significant differences in concentrations of 12(S)LOX in different groups of patients. CONCLUSIONS: Our results suggest that 12(S)LOX is relevant in prostate cancer; however, further study should include a larger, more select group of men with prostate cancer.

Parameters of antioxidant barrier in different histopathologic types of pediatric cancers.

AIM OF THE STUDY: The goal of this study was to evaluate the activities of erythrocyte superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and the levels of glutathione (GSH) and ischemia-modified albumin (IMA), as potential markers in different histopathologic types of pediatric neoplasms. No studies on this subject have been reported to date. MATERIAL AND METHODS: SOD, GSH-Px, GSH, and IMA were measured before oncologic treatment in 129 children with neuroblastoma (NB), soft tissue sarcomas (STS), brain tumors, Hodgkin's disease (HD), and acute leukemias, and in 30 healthy controls. RESULTS: The statistical significance of SOD was observed in patients with brain tumors (median 1840.2 U/g Hb, p = 0.0500). The level of GSH was significantly higher in patients with NB (median 6.38 U/g Hb, p = 0.0031) and leukemias (5.16 U/g Hb, p = 0.0200). IMA was statistically significant in cases of STS, NB, and leukemias compared to healthy children (p = 0.0244, p = 0.0069, and p = 0.0000, respectively). The activity of GSH-Px was not statistically significant. CONCLUSIONS: The antioxidant barrier in all types of pediatric cancers is disturbed. None of the measured parameters was specific enough to represent a reliable marker for any particular histopathologic type of children's neoplasm.

The antioxidant status and response to therapy in children with soft tissue sarcomas and neuroblastoma.

BACKGROUND: Antioxidant systems in cells maintain the proper homeostasis of reactive oxygen species, which at high concentrations can induce carcinogenesis. The aim of this study was to evaluate the serum levels of ischemia-modified albumin (IMA), erythrocyte superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) as markers for prognosis in children with neuroblastoma (NB) and soft tissue sarcomas (STS), two cancer types for which reliable prognostic factors are needed. PROCEDURE: SOD, GSH-Px, and IMA were measured before and during responses to therapy assessment in 99 children with NB and STS and in 30 healthy controls. RESULTS: There were no statistically significant differences in the erythrocyte SOD and GSH-Px activities between the patients with cancer and healthy controls. The levels of IMA in patients with STS and NB were found to be significantly higher than in the controls (P = 0.0013; P = 0.0066, and 0.0164, respectively). Decreased activities of SOD and GSH-Px were found in all patients with poor-responding (PRS) cancers and decreased SOD activity was found in patients with PRS NB. An increase in GSH-Px was observed in patients with good-responding (GR) NB. All patients with GR cancers demonstrated higher SOD and GSH-Px activities than patients with PRS cancers. CONCLUSIONS: While determining the levels of specific antioxidants as antioxidant-barrier parameters in children with cancer may be valuable in predicting therapeutic responses as well as outcomes, additional studies are required.

Effects of renal-specific oral supplementation in malnourished hemodialysis patients.

BACKGROUND: Protein-energy malnutrition is still a problem in patients with chronic renal failure, especially during replacement renal therapy. The chronic inflammatory status in these patients intensifies the malnutrition, as well as making treatment more complicated. The aim of the present study was to estimate the influence of oral supplementation on the nutritional status of malnourished hemodialysis (HD) patients depending on the existence of an inflammatory state. METHODS: To study the influence of oral supplementation on nutrition status, 30 HD patients with protein-energy malnutrition characteristics and 25 well-nourished HD patients were enrolled in the study. Malnourished HD patients were prescribed Renilon 7.5 at an oral intake dose of 125 mL twice a day for 3 months. The nutritional status was characterized based on body mass index, Subjective Global Assessment score, serum albumin and prealbumin concentrations. The intensity of the inflammatory state was determined by C-reactive protein and interleukin-6. Serum concentrations of leptin and adiponectin were also measured. RESULTS: After 3 months of supplementation, malnourished patients had an increase in prealbumin, albumin, and leptin concentrations. No statistically significant differences were observed between patients lacking inflammation and those with inflammation. CONCLUSIONS: The results indicate an improvement in the nutritional status of HD patients who were prescribed an oral supplementation. Furthermore, patients with inflammatory state characteristics also benefited from Renilon 7.5 treatment.

Very long half-life plasminogen activator inhibitor type 1 reduces bleeding in a mouse model.

OBJECTIVE: To investigate the potential for the future clinical use of a very long half-life plasminogen activator inhibitor type 1 (VLHL PAI-1) as a haemostatic agent. MATERIALS AND METHODS: We developed a VLHL PAI-1 (half-life >700 h) recombinant mutant of PAI-1 and assessed VLHL PAI-1 for its ability to inhibit fibrinolysis in vitro using human, rabbit, mouse and rat blood. Fibrin clot lysis time, monitored by thromboelastometry, was determined at various concentrations of VLHL PAI-1. Also, we determined total bleeding time and total blood loss of control, VLHL PAI-1-, tissue-type plasminogen activator (tPA)- and tPA + VLHL PAI-1-treated mice. RESULTS: Using a thromboelastometer, mouse blood was most similar to human blood in its coagulation and fibrinolytic characteristics. We evaluated the affect of VLHL PAI-1 on haemostasis using the mouse model and showed that VLHL PAI-1 is an effective inhibitor of fibrin clot degradation. It reduced time of bleeding and total blood loss. CONCLUSION: VLHL PAI-1 may provide an important physiological mechanism to protect clots from premature dissolution in surgical and trauma settings.

The effect of N-acetylcysteine on blood pressure and markers of cardiovascular risk in non-diabetic patients with chronic kidney disease: a placebo-controlled, randomized, cross-over study.

BACKGROUND: Cardiovascular complications in patients with chronic kidney disease (CKD) are frequent. They show increased cardiovascular mortality and morbidity attributable to accumulation of several risk factors; e.g., hypertension, oxidative stress and elevated plasma homocysteine concentration. Despite recent progress in their management, there is still no optimal therapy that can stop progression of CKD and decrease cardiovascular outcome in these patients. Antioxidants, e.g., N-acetylcysteine (NAC), have been suggested as a promising medicament in this field. MATERIAL/METHODS: In a placebo-controlled, randomized, two-period cross-over study we evaluated the influence of eight weeks of NAC therapy (1200 mg/day) added to pharmacological renin-angiotensin system blockade on ambulatory blood pressure and surrogate markers of cardiovascular risk and injury in 20 non-diabetic patients with albuminuria [30-915 mg per creatinine mg] and normal or slightly decreased kidney function [eGFR 61-163 ml/min]. After eight weeks run-in period during which the therapy using angiotensin converting enzyme inhibitors and/or angiotensin receptor blockers was settled, patients were randomly assigned to one of two treatment sequences: NAC/washout/placebo or placebo/washout/NAC. RESULTS: No significant changes in blood pressure, albuminuria and homocysteine plasma level were observed. CONCLUSIONS: NAC had no effect on blood pressure and surrogate markers of cardiovascular injury in non-diabetic patients with CKD.

Adipokines, endothelial dysfunction and nutritional status in peritoneal dialysis patients.

OBJECTIVE: Adipokines such as leptin and adiponectin are adipocyte-specific secretory proteins that play important roles in the metabolic regulation of body weight, insulin resistance and cardiovascular complications. The relationship between the malnutrition-inflammation complex syndrome and high levels of some adipokines in peritoneal dialysis (PD) patients is still unclear. An association between high body mass index (BMI) and improved survival in PD patients has also been proposed. The purpose of this study was to investigate the levels of plasma adipokines and inflammation and oxidative stress markers in overweight and normal weight PD patients. MATERIAL AND METHODS: Thirty PD patients (12 M, 18 F; mean age 57.3 ± 16.6 years) were examined and 23 healthy volunteers were included as a control group. The levels of high-sensitivity C-reactive protein (hsCRP), tumour necrosis factor-α, interleukin-6, leptin, the leptin receptor, adiponectin, malondialdehyde/4-hydroxynonenal, oxidized low-density lipoprotein, carbonyl groups and asymmetric dimethylarginine (ADMA) were measured in both groups. The nutritional status of each patient was determined by albumin levels, BMI, percentage of body fat (%F), lean body mass (LBM) and the Subjective Global Assessment (SGA) score. The adequacy of dialysis was estimated by weekly Kt/V measurements. RESULTS: According to the seven-point SGA scores and the albumin levels, the nutrition status of 15 patients was good (6-7 points), while 15 patients were mildly malnourished (3-5 points). The concentrations of hsCRP, leptin and adiponectin were statistically higher in the PD group than in the control group (p < 0.05). Markers of oxidative stress and inflammation were also higher in the PD group. The adiponectin level was inversely correlated with %F and BMI (Spearman's R = -0.3, p ≤ 0.05) and positively correlated with hsCRP level (R = -0.4). The level of leptin was positively correlated with %F, BMI and LBM (R = 0.4, p ≤ 0.05). Patients with normal BMI values had lower leptin concentrations (50.2 vs 242.8 µg/l) and higher adiponectin levels (30.0 vs 20.3 µg/ml) than overweight patients. The statistical analysis indicated that there were no differences in oxidative stress, inflammation and ADMA concentration between the lean and overweight PD patients. CONCLUSION: The nutritional status of lean and overweight patients was comparable. Signs of malnutrition were detected in both groups. The severity of chronic inflammation and oxidative stress were not related to BMI in PD patients.

The serum profile of adipokines in overweight patients with metabolic syndrome.

INTRODUCTION: Obesity is a disease that brings several complications and increases the risk of other diseases like metabolic syndrome, diabetes mellitus type 2, or coronary heart disease. Disturbances in secretion of adipokines caused by obesity have an influence on the development of metabolic complications. The aim of this study was an investigation of adipokines profile in overweight or obese people with metabolic syndrome in comparison to overweight/obese patients without metabolic syndrome. MATERIAL AND METHODS: The studied groups consisted of 38 obese or overweight patients without metabolic syndrome (nonMS) and 17 with recognized metabolic syndrome (MS), according to International Diabetes Federation (IDF) criteria. All individuals underwent anthropometrical and blood-pressure examination as well as biochemical analyses such as: serum concentrations of glucose, insulin, adiponectin, resistin, leptin, TNF-alpha, IL-6, hs-CRP, total cholesterol, HDL, and triglycerides. RESULTS: A significantly lower concentration of adiponectin, and a higher concentration of IL-6, was observed in patients with metabolic syndrome (MS) in comparison to nonMS. Moreover, higher concentrations of hs-CRP and TNF-alpha were observed in patients with metabolic syndrome. CONCLUSIONS: A decreased concentration of adiponectin in obese people is an early predictor of metabolic syndrome. A low adiponectin level could be a marker of high risk of cardiovascular disease in obese patients. (Pol J Endocrinol 2010; 61 (1): 36-41).

Adipose tissue activity in relation to overweight or obesity.

INTRODUCTION: Obesity is associated with a number of diseases resulting from the excessive amount of adipose tissue. The aim of this study was to investigate the correlation between the quantity of adipose tissue and the prevalence of metabolic disturbances, and the concentration of adipokines and proinflammatory cytokines in obese or overweight patients. MATERIAL AND METHODS: Fifty-five middle-aged subjects with body mass index (BMI) > 25 kg/m(2) took part in this study. Twenty-three healthy people with normal BMI formed the control group. Twenty-one people from the study group were on a low-calorie diet. All subjects underwent anthropometric assessment, laboratory investigations, and blood-pressure examination. RESULTS: Patients with obesity or overweight, in comparison to those with normal BMI, showed insulin resistance and a higher concentration of high sensitivity C-reactive protein (hs-CRP), plasminogen activator inhibitor 1 (PAI-1), and interleukin 6 (Il-6). The concentration of adiponectin was significantly lower in this group. The patients on the low-calorie diet had significantly lower concentrations of leptin when compared to other obese people; moreover, a trend towards decreased hs-CRP concentration was seen. A significant positive correlation between leptin and hs-CRP was observed. The serum concentration of adiponectin was inversely correlated with that of TNF-alpha, IL-6, hs-CRP, and PAI-1. CONCLUSIONS: The results of this study may suggest the beneficial impact of a low-calorie diet on the slowing down of inflammatory processes. The observed negative correlation between the concentrations of adiponectin and inflammatory cytokines may confirm the anti-inflammatory activity of this adipokine.

Do human lipoxygenases have a PDZ regulatory domain?

Human lipoxygenases and products of their catalytic reaction have a well established connection to many human diseases. Despite their importance in inflammation, cancer, cardiorenal and other ailments the drug development is impaired by the lack of structural details to understand their intricate specificity and function in molecular and cellular signaling. The major effort so far has been directed towards understanding the determinants of their specificity and inhibition of their active site with the iron cofactor. Their structure is believed to consist of only two domains: one regulatory - a beta-sandwich, important for membrane binding, and one, mostly helical, catalytic domain. Although recently published cohort studies on single nucleotide polymorphism and occurrence of diseases, SAXS analysis and new biochemical data throw new light on lipoxygenase suggesting symbiosis of regulatory functions with an allosteric mechanism and more flexible structure than anticipated. The goal of this brief review is to direct an attention to the structural features of an anticipated topology and stimulate discussion/research to prove or disapprove our hypothesis that lipoxygenases may possess about approximately 110 amino acids PDZ-like fragments of functional importance. If they do have a second regulatory domain, it might help to explain their association with other molecules, role in signaling pathways and present a new avenue to explore the regulation of their behavior, and thus intervention in the course of diseases.

Increased Trimethylamine N-Oxide Is Not Associated with Oxidative Stress Markers in Healthy Aged Women.

Increased plasma trimethylamine N-oxide (TMAO) levels have been associated with cardiovascular diseases (CVD). L-carnitine induces TMAO elevation in human blood, and thus, it has been suggested as developing atherosclerosis. The aim of this study was to determine the relation between selected markers of oxidative stress and plasma TMAO concentration induced by L-carnitine supplementation for 24 weeks in healthy aged women. Twenty aged women were supplemented during 24 weeks with either 1500 mg L-carnitine-L-tartrate (n = 11) or isonitrogenous placebo (n = 9) per day. Fasting blood samples were taken from antecubital vein. L-carnitine supplementation induced an increase in TMAO, but not in γ-butyrobetaine (GBB). Moreover, there were no significant changes in serum ox-LDL, myeloperoxidase, protein carbonyls, homocysteine, and uric acid concentrations due to supplementation. Significant reduction in white blood cell counts has been observed following 24-week supplementation, but not attributable to L-carnitine. Our results in healthy aged women indicated no relation between TMAO and any determined marker of oxidative stress over the period of 24 weeks. At the same time, plasma GBB levels were not affected by L-carnitine supplementation. Further clinical studies of plasma GBB level as a prognostic marker are needed.

Triple pharmacological blockade of the renin-angiotensin-aldosterone system in nondiabetic CKD: an open-label crossover randomized controlled trial.

BACKGROUND: Agents inhibiting the renin-angiotensin-aldosterone (RAAS) system have an important role in slowing the progression of chronic kidney disease. We evaluated the hypothesis that the addition of an aldosterone receptor antagonist to an angiotensin-converting enzyme (ACE) inhibitor and angiotensin II type 1 (AT-1) receptor blocker (ARB) (triple RAAS blockade) may provide an additional benefit compared with an ACE inhibitor and ARB (double RAAS blockade). DESIGN: Randomized open controlled crossover study. SETTING & PARTICIPANTS: 18 whites (7 women, 11 men) from the Outpatient Department of Nephrology with chronic nondiabetic proteinuric kidney diseases, mean age 42.4 +/- 1.9 years (SEM). INTERVENTIONS: In the 8-week run-in period, all participants received the ACE inhibitor cilazapril (5 mg), the ARB telmisartan (80 mg), and the diuretic hydrochlorothiazide (12.5 mg) as double RAAS blockade to achieve the target blood pressure of less than 130/80 mm Hg. Participants were then randomly assigned to 2 treatment sequences, either the addition of spironolactone (25 mg) (triple RAAS blockade) through 8 weeks followed by double RAAS blockade through 8 weeks (sequence 1) or double RAAS blockade followed by triple RAAS blockade (sequence 2). MAIN OUTCOME MEASURES: 24-hour urine protein excretion (primary end point) and markers of tubular injury and fibrosis (secondary end points). Analysis was performed using analysis of variance for repeated measurements. RESULTS: At baseline, mean serum creatinine level was 1.16 +/- 0.09 mg/dL (103 +/- 8 micromol/L), estimated glomerular filtration rate was 107.8 mL/min (95% confidence interval, 93 to 140.9 [1.8 mL/s; 95% confidence interval, 1.55 to 2.35; Cockcroft-Gault formula), and 24-hour mean proteinuria was 0.97 +/- 0.18 g. Mean urine protein excretion was 0.7 g/24 h (95% confidence interval, 0.48 to 0.92) less after triple RAAS blockade than after double RAAS blockade (P = 0.01), without change in blood pressure. Urine excretion of N-acetyl-beta-d-glucosaminidase (P = 0.02) and amino-terminal propeptide of type III procollagen (P = 0.05) also significantly decreased. Potassium levels increased significantly after triple therapy (P = 0.02). However, no patient was withdrawn because of adverse effects. LIMITATIONS: Absence of blinding, small sample size, short treatment period, absence of histological assessment. CONCLUSIONS: Administration of an aldosterone receptor antagonist in addition to double RAAS blockade with an ACE inhibitor and ARB may slow the progression of chronic kidney disease. Additional studies are necessary to confirm this result.

Uric acid excretion and dopamine-induced glomerular filtration response in patients with IgA glomerulonephritis.

BACKGROUND/AIM: It is unknown to what extent uric acid (UA) may affect vessel function and participate in tubulointerstitial damage. We examined the relationship between intrarenal vessel function and serum UA and its excretion in association with urinary N-acetyl-beta-D-glucosaminidase (NAG). METHODS: In 50 IgA patients (mean age 34.7 +/- 9.3 years) and 15 controls (mean age 33.5 +/- 6.9 years) with a creatinine clearance of 99.4 +/- 21.6 and 118.1 +/- 17.2 ml/min, respectively, the renal vascular function was estimated based on the dopamine-induced glomerular filtration response (DIR; see text). The DIR was measured using two 120-min creatinine clearance values (before and after intravenous administration of 2 g/kg/min dopamine). Serum UA, triglycerides and cholesterol and urinary NAG (24 h) and protein and UA excretion were measured. RESULTS: Patients with IgA nephropathy versus controls: DIR 8.80 +/- 6.6 vs. 12.83% (p < 0.01), NAG 7.25 +/- 3.30 vs. 4.69 +/- 1.12 U/g creatinine (p < 0.01), and fractional UA excretion 7.80 +/- 2.20 versus 6.29 +/- 1.80% (p < 0.01). A negative correlation between DIR and NAG was found; regression analysis showed a more prominent relationship in the patients (NAG = 9.99 - 0.29x DIR) than in the controls (NAG = 5.50 - 0.06x DIR). UA and urate excretion and NAG in the patients correlated with DIR (r = -0.39, p < 0.02; r = -0.29, p < 0.04, and r = 0.59, p < 0.001, respectively). Multivariate analysis showed an association of DIR (R(2) = 0.39) with NAG but not with proteinuria and UA and UA excretion; the NAG excretion (R(2) = 0.56) correlated significantly with UA and DIR. CONCLUSION: It is suggested that UA plays a role, associated with tubular dysfunction, in the regulation of intrarenal vessel function.

The effect of N-acetylcysteine on proteinuria and markers of tubular injury in non-diabetic patients with chronic kidney disease. A placebo-controlled, randomized, open, cross-over study.

BACKGROUND: Inhibition of the renin-angiotensin-aldosterone system with angiotensin-converting enzyme inhibitors (ACEI) and/or angiotensin II subtype 1 receptor antagonists (ARB) constitutes a strategy in the management of patients with chronic kidney disease. There is still no optimal therapy which can stop the progression of chronic kidney disease. Antioxidants such as N-acetylcysteine (NAC) have been reported as a promising strategy in this field. METHODS: In a placebo-controlled, randomized, open, 2-period cross-over study, we evaluated the influence of NAC (1,200 mg/day) added to renin-angiotensin-aldosterone system blockade on proteinuria and surrogate markers of tubular injury and renal fibrosis in 20 non-diabetic patients with proteinuria (0.4-6.36 g/24 h) with normal or decreased kidney function (estimated glomerular filtration rate 61-163 ml/min). Subjects entered the 8-week run-in period during which the therapy using ACEI and/or ARB was established with blood pressure below 130/80 mm Hg. Next, patients were randomly assigned to 1 of 2 treatment sequences: NAC/washout/placebo or placebo/washout/NAC. Clinical evaluation and laboratory tests were performed at the randomization point and after each period of the study. RESULTS: No significant changes in laboratory tests were observed. CONCLUSION: NAC had no effect on proteinuria, surrogate markers of tubular injury or renal fibrosis in non-diabetic patients with chronic kidney disease.

Serum concentration of amino acids versus nutritional status in hemodialysis patients.

OBJECTIVE: The aim of this study was to evaluate the correlation between the serum concentration of amino acids (AAs) and nutritional status in hemodialysis (HD) patients. METHODS: This study was performed in 22 HD patients dialyzed for 10 to 288 months, and in a control group of 20 healthy volunteers. Nutritional status was determined by the subjective global assessment method and by measuring albumin concentration. Body composition was determined using the parameters of body mass index, and the percentage of body fat and lean body mass (as measured by the near-infrared method). We measured C-reactive protein (CRP) as a marker of inflammatory status. Serum concentrations of 20 AAs were measured by precolumn orthophtalaldehyde derivatization, applying high-performance liquid chromatography (Hitachi-Merck HPLC, Tokyo, Japan) equipped with a C-18 reversed-phase column and a methanol/acetate buffer gradient. RESULTS: Thirteen of 22 (59%) patients were of good nutritional status, and 9/22 (41%) were malnourished, including 1 person with severe malnutrition. In dialyzed patients compared with control subjects, a decreased concentration of essential and nonessential AAs was observed (P < .05). Concentrations of the majority of studied AAs (16 out of 20) were lower in patients dialyzed for a period >2 years, compared with patients dialyzed for a shorter time. The ratio of branched-chain amino acids (BCAAs) to aromatic AAs was lower in the dialyzed group compared with control subjects. This ratio was also lower in patients dialyzed longer compared with patients dialyzed for <2 years. No correlation between the concentration of some AAs and CRP level or dialysis adequacy was observed. In the malnourished group, an insignificantly lower concentration of some essential AAs (lysine, leucine, isoleucine, valine, and threonine), and a significantly higher (P = .04) concentration of CRP, were observed. CONCLUSION: Despite quite good nutritional status, dialyzed patients present abnormalities in their AA profiles. Moreover, a significant decrease of BCAA concentration is related to calorie-protein malnutrition, inflammation, and a long period of hemodialysis.

l-Carnitine Supplementation in Older Women. A Pilot Study on Aging Skeletal Muscle Mass and Function.

Skeletal muscle wasting, associated with aging, may be regulated by the inflammatory cytokines as well as by insulin-like growth factor 1 (IGF-1). l-carnitine possesses anti-inflammatory properties and increases plasma IGF-1 concentration, leading to the regulation of the genes responsible for protein catabolism and anabolism. The purpose of the present study was to evaluate the effect of a 24-week l-carnitine supplementation on serum inflammatory markers, IGF-1, body composition and skeletal muscle strength in healthy human subjects over 65 years of age. Women between 65 and 70 years of age were supplemented for 24 weeks with either 1500 mg l-carnitine-l-tartrate or an isonitrogenous placebo per day in a double-blind fashion. Before and after the supplementation protocol, body mass and composition, as well as knee extensor and flexor muscle strength were determined. In the blood samples, free carnitine, interleukin-6, tumor necrosis factor-α, C-reactive protein and IGF-1 were determined. A marked increase in free plasma carnitine concentration was observed due to l-carnitine supplementation. No substantial changes in other parameters were noted. In the current study, supplementation for 24 weeks affected neither the skeletal muscle strength nor circulating markers in healthy women over 65 years of age. Positive and negative aspects of l-carnitine supplementation need to be clarified.

PAI-1 induces cell detachment, downregulates nucleophosmin (B23) and fortilin (TCTP) in LnCAP prostate cancer cells.

Plasminogen activator inhibitor (PAI-1) is an anticancer agent that inhibits plasmin driven proteolysis, limiting angiogenesis and metastasis. In low concentrations it could induce cancer cell motility by interacting with urokinase (uPA), its receptor (uPAR), vitronectin and integrins. Active PAI-1 binds to uPA forming a complex with uPAR, while the latent form of PAI-1 does not. PAI-1 is found in both forms in the circulation. It is not clear which form acts as an anticancer agent and how it interacts with malignant cells. To investigate how these forms reduce angiogenesis or metastasis, we have created PAI-1 cysteine mutants in the active conformation (VLHL PAI-1) with an extended half-life that reaches approximately 700 h and its R369A mutant, which has an active conformation but cannot bind to uPA (VLHLNS PAI-1). Both VLHL PAI-1s convert into the latent form when treated with a reducing agent (DTT) that breaks disulfide bridges. Unexpectedly, during routine investigation of LnCAP cell proliferation, we have found that cells detach from the culture vessels regardless of PAI-1 conformation or activity. Further investigation showed that treatment of cancer cells with VLHL PAI-1 downregulated nucleophosmin, while all forms of PAI-1 downregulated fortilin. These two proteins are implicated in important cellular processes (cell growth, cell cycle, malignant transformation). This suggests that PAI-1, in addition to its well-known anticancer properties, plays an important role in cell signaling. We hope that by exploring PAI-1's structure and function we might be able to understand and separate the different effects of PAI-1 on cancer cells and develop more effective therapeutic strategies in cancer treatment.

Highly stable plasminogen activator inhibitor type one (VLHL PAI-1) protects fibrin clots from tissue plasminogen activator-mediated fibrinolysis.

Plasminogen activator inhibitor-1 (PAI-1) is the major specific inhibitor of tissue-type plasminogen activator (tPA) which mediates fibrin clot lysis through activation of plasminogen. Wild-type-PAI-1 (wPAI-1) is rapidly converted to the latent form (half-life of approximately 2 h) and loses its ability to inhibit tPA. We developed a very long half-life PAI-1 (VLHL PAI-1), a recombinant protein with a half-life >700 h compared with wPAI-1. In this study, VLHL PAI-1 was assessed for its ability to inhibit clot lysis in vitro. Clot formation was initiated in normal plasma supplemented with tPA by the addition of either tissue factor or human recombinant FVIIa. Clot lysis time, monitored turbidimetrically in a microtiter plate reader, was determined at various concentrations of wPAI-1 and VLHL PAI-1. Both wPAI-1 and VLHL PAI-1 caused a significant increase in clot lysis time, although the latter was somewhat less effective at lower concentrations. The VLHL PAI-1, but not wPAI-1, maintained its anti-fibrinolytic activity after preincubation overnight at 37 degrees. These studies demonstrate that VLHL PAI-1 is an effective inhibitor of fibrin clot degradation. Due to the high stability of VLHL PAI-1 compared with wPAI-1, this novel inhibitor of tPA-mediated fibrinolysis may have therapeutic applications for treating surgical and trauma patients when used directly or in conjunction with the procoagulant recombinant FVIIa.