Ladislav Trefil

Influence of chromium-enriched yeast on blood glucose and insulin variables, blood lipids, and markers of oxidative stress in subjects with type 2 diabetes mellitus

The aim of this study was to determine the effect of chromium (Cr)-enriched yeast on blood glucose and insulin variables, blood lipids, and blood markers of oxidative stress in persons with type 2 diabetes mellitus (median duration: 3.0 yr). Thirty-six subjects (9 men, 27 women; mean age: 61.3 yr; mean body mass index: 34.33 kg/m2) were supplemented with 400 μg Cr/d as Cr-enriched yeast (n=19) or placebo (n=17) for 12 wk in a randomized, double-blind study. The most interesting results were obtained by comparison of the change in the placebo group to the change in the Cr group. The Cr group showed a significantly greater increase in serum Cr compared to the placebo group (p<0.05). Supplementation with Cr-enriched yeast was associated with a significant decrease in fasting serum glucose compared to placebo (p<0.01). Blood markers of oxidative stress glutathione peroxidase activity and levels of reduced glutathione were essentially unchanged in the Cr group after 12 wk, but decreased significantly in the placebo group (p<0.05, p<0.01, respectively), Serum HbA1c and glycated protein (fructosamine) were essentially unchanged in the Cr group, whereas HbA1c tended to increase in the placebo group (from 6.94% to 7.11%). Fasting serum insulin decreased in both groups, with a greater tendency in the Cr group (−16.5% vs −9.5%). These data suggest that supplementation of well-controlled type 2 diabetics with Cr-enriched yeast is safe and can result in improvements in blood glucose variables and oxidative stress.

Parameters of oxidative stress in children with Type 1 diabetes mellitus and their relatives

Oxidative stress (OS) plays an important role in the pathogenesis of Type 1 diabetes mellitus (DM). The aim of the study was to compare OS parameters in diabetic children and their first-degree relatives. Fifty diabetic children from the West Bohemian Region were examined as well as their 32 siblings (12 Boys and 20 girls) and 65 of their parents during a period of 6 months. Thirty healthy sex- and age-matched children studied before planned surgeries were normal controls for children, 40 healthy adult volunteers were controls for parents. OS parameters were evaluated in all participants of the study (superoxide dismutase, SOD; glutathione peroxidase, GSHPx; plasma antioxidant capacity, AOC; reduced glutathione, GSH; and malondialdehyde, MDA) and also Type 1 DM-associated antibodies (ICA and GADA). The results in diabetic children showed significantly lower GSHPx and AOC and increased MDA when compared with healthy children. Similar findings were found in their siblings but without statistical significance. It is consequently evident that decreased antioxidative protection and simultaneous free radical (FR) overproduction occur in diabetic children and that there is a similar, but not significant, tendency in their siblings. The findings warrant reducing OS in diabetic children and postponing disease onset in susceptible relatives.

Brain Natriuretic Peptide and N-Terminal proBNP in Chronic Haemodialysis Patients

Background: Brain natriuretic peptide (BNP) and N-terminal proBNP (NT-proBNP) are released into circulation as a result of congestive heart failure (HF). As HF and water overload are frequent complications in haemodialysis (HD) patients, we decided to study the levels of BNP and NT-proBNP and their changes during HD. Methods: BNP and NT-proBNP levels were determined in 94 HD patients before and after a regular 4-h HD. We followed changes in these peptides during HD depending on age, sex, HF (NYHA classification and left ventricular ejection fraction [LVEF]), duration on HD, presence of hypertension, coronary artery disease, type of membrane used for HD [low-flux (LFx) or high-flux (HFx)] and body mass change during HD. Furthermore, patients´ basic medication and creatinine levels and presence of diabetes mellitus were monitored. Results: Respectively,94% and 100% of the patients had pre-dialysis concentrations of BNP and NT-proBNP above the cut-off values for HF. The marker levels correlated significantly both before and after HD (r = 0.903 and 0.888, respectively, p < 0.001). BNP levels significantly decreased (p < 0.0001), whereas NT-proBNP significantly increased (p < 0.0001) during HD on LFx membranes. HD on HFx membranes caused greater decrease of BNP (compared to LFx membranes, p < 0.001), but also a decrease of NT-proBNP (p < 0.001).We did not find any significant differences in marker levels for HF and non-HF patients (NYHA classification). However, both peptides reached higher levels in the group with LVEF ≤50% (p < 0.001 for both peptides). Body mass change during HD negatively correlated only with the change of NT-proBNP (r = –0.27, p < 0.05). In the multiple regression model, the change of both peptides during HD was significantly influenced by membrane type (p = 0.003 for BNP and p = 0.001 for NT-proBNP). NT-proBNP change during HD was further significantly influenced by LVEF (p = 0.012), sex (p = 0.002) and duration on HD (p = 0.006). Conclusions: Both BNP and NT-proBNP levels were significantly increased in HD patients prior to dialysis. The change in concentrations of both peptides during HD is influenced by membrane type. HD probably triggers increased production of both peptides and this increase is emphasized by impaired LVEF. This fact can be clinically observed only on NT-proBNP levels, because BNP levels are biased by significant removal of this protein during HD.

Asymmetric dimethylarginine in hemodialysis, hemodiafiltration, and peritoneal dialysis.

Asymmetric dimethylarginine (ADMA) is a mediator of endothelial dysfunction. Production and elimination of ADMA may be affected by the type of renal replacement therapy used and oxidative stress. Plasma ADMA, advanced glycation end products (AGE), and homocysteine were assessed in 59 subjects: 20 hemodialysis (HD) patients, 19 patients undergoing peritoneal dialysis (PD), and 20 controls. Results were compared between the groups. The effect of 8 weeks of HD and high-volume predilution hemodiafiltration (HDF) was compared in a randomized study. HD patients showed higher ADMA (1.20 [0.90-1.39 micromol/L]) compared to controls (0.89 [0.77-0.98], P < 0.01), while ADMA in PD did not differ from controls (0.96 [0.88-1.28]). AGE and homocysteine were highest in HD, lower in PD (P < 0.01 vs. HD), and lowest in controls (P < 0.001 vs. HD and PD). PD patients had higher residual renal function than HD (P < 0.01). The decrease in ADMA at the end of HD (from 1.25 [0.97-1.33] to 0.66 [0.57-0.73], P < 0.001) was comparable to that of HDF. Switching from HD to HDF led to a decrease in predialysis homocysteine level in 8 weeks (P < 0.05), while ADMA and AGE did not change. Increased ADMA levels in patients undergoing HD, as compared to PD, may be caused by higher oxidative stress and lower residual renal function in HD. Other factors, such as diabetes and statin therapy, may also be at play. The decrease in ADMA at the end of HD and HDF is comparable. Switching from HD to HDF decreases in 8 weeks the predialysis levels of homocysteine without affecting ADMA.

The Effect of Intravenous Iron on Oxidative Stress in Hemodialysis Patients at Various Levels of Vitamin C

Background/Aims: Vitamin C levels decrease during hemodialysis (HD), which deteriorates antioxidant defense. Vitamin C may also act pro-oxidatively, via reduction in Fe(III). We sought to determine whether intravenous iron (Fe_iv)-induced oxidative stress differs in HD patients with low and physiological vitamin C levels and whether intravenous vitamin C (C_iv) administration during HD would change the response to Fe_iv. Patients and Methods: Twenty patients with vitamin C deficiency (median 15.7 µmol/l, range 8.0–22.7) received Fe_iv (100 mg iron sucrose between 150 and 180 min of HD). After 4 weeks of oral supplementation, the levels of vitamin C were comparable with those of controls (60.1 µmol/l, range 47.4–70.9). Patients were subsequently treated with (1) Fe_iv, (2) Fe_iv and continuous 2 mg/min C_iv throughout HD, (3) saline (S), and (4) S+C_iv. Plasma thiobarbituric acid reacting substances (TBARS) and vitamin C were assessed before, during and after FE_iv(S), and 15, 30 and 60 min after infusion. Results: Fe_iv induced a comparable rise in TBARS in patients with vitamin C deficiency (before Fe_iv, 1.9 µmol/l, range 1.4–1.9; after Fe_iv, 2.6 µmol/l, range 2.3–2.9; p < 0.01) and in those with normal vitamin C (before Fe_iv, 1.9 µmol/l, range 1.7–2.1; after Fe_iv, 2.6 µmol/l, range 2.5–2.9; p < 0.01). Fe_iv+C_iv resulted in a greater increase in TBARS (after Fe_iv, 3.1 µmol/l, range 2.8–3.2) compared with Fe_iv (p < 0.01). Conclusion: Iron sucrose-induced oxidative stress is comparable in HD patients with vitamin C deficiency and in those with normal vitamin C. We documented a pro-oxidative effect of vitamin C during Fe_iv+C_iv administration.

Effect of Folic Acid, Betaine, Vitamin B6, and Vitamin B12 on Homocysteine and Dimethylglycine Levels in Middle-Aged Men Drinking White Wine

Moderate regular consumption of alcoholic beverages is believed to protect against atherosclerosis but can also increase homocysteine or dimethylglycine, which are putative risk factors for atherosclerosis. We aimed (1) to investigate the effect of alcohol consumption on vitamins and several metabolites involved in one-carbon metabolism; and (2) to find the most effective way of decreasing homocysteine during moderate alcohol consumption. Methods: Male volunteers (n = 117) were randomly divided into five groups: the wine-only group (control, 375 mL of white wine daily for one month) and four groups combining wine consumption with one of the supplemented substances (folic acid, betaine, and vitamins B12 or B6). Significant lowering of homocysteine concentration after the drinking period was found in subjects with concurrent folate and betaine supplementation. Vitamin B12 and vitamin B6 supplementation did not lead to a statistically significant change in homocysteine. According to a multiple linear regression model, the homocysteine change in the wine-only group was mainly determined by the interaction between the higher baseline homocysteine concentration and the change in dimethylglycine levels. Folate and betaine can attenuate possible adverse effects of moderate alcohol consumption. Dimethylglycine should be interpreted together with data on alcohol consumption and homocysteine concentration.

Serum citrulline levels as a marker of enterocyte function in patients after allogeneic hematopoietic stem cells transplantation – a pilot study

Background Citrulline is an amino acid produced by enterocytes. Serum citrulline concentration has been proposed as a marker of enterocyte mass and function. Our study focused on evaluation of citrulline levels in patients with diarrhea related to toxic intestinal damage (mucositis), intestinal graft versus host disease (GVHD), and other etiology of diarrhea (e.g., dysmicrobia) after allogeneic stem cells transplantation (SCT). Material/Methods This was a prospective study in 11 adults (18 blood samples) with diarrhea developed after allogeneic SCT in 4/2011–1/2012 compared to twenty healthy control samples. Results The median (interquartile range) of citrulline levels was significantly lower in the transplanted patients group compared to healthy controls: 9.3 (3.62–15.38) vs. 33.3 (26.82–36.23) μmol/L, p<0.0001. The median values of citrulline in patients with post-transplant toxic intestinal mucositis (n=8, days 1–22 post-transplant) vs. intestinal GVHD (n=7, day 43–142) vs. other etiology of diarrhea (e.g., dysmicrobia) (n=3, day 120–127) were: 9.55 (2.95–12.03) vs. 5 (3.85–9.05) vs. 15.6 (15.45–18.3) μmol/L resp. Serum citrulline levels were significantly higher in other (eg, dysmicrobic) etiology of diarrhea in comparison with mucositis (p=0.0336) and GVHD (p=0.0152). Conclusions Citrulline levels are very low shortly after the myeloablative FLU/MEL or BuCY2 conditioning allogeneic SCT due to the toxic intestinal damage. Significantly low levels of citrulline were also in patients with intestinal GVHD later on. Other observations in larger groups of patients are necessary before any specific recommendation for citrulline levels monitoring in intestinal GVHD can be made.

Intraperitoneal IL-6 Signaling in Incident Patients Treated with Icodextrin and Glucose Bicarbonate/Lactate–Based Peritoneal Dialysis Solutions

♦ Objective: In this study, we compared the activity of interleukin-6 (IL-6), a marker of ongoing peritoneal inflammation and biocompatibility, and its other signaling components, the soluble IL-6 receptor (sIL-6R) and soluble Gp130 (sGp130), in peritoneal effluent from patients treated with icodextrin-based (E) peritoneal dialysis (PD) solution and glucose-based bicarbonate/lactate–buffered (P) solution. ♦ Methods: Using baseline peritoneal ultrafiltration capacity, 33 stable incident PD patients were allocated either to P only (n = 20) or to P plus E for the overnight dwell (n = 13). We used ELISA to determine IL-6, sIL-6R, and sGp130 in timed overnight effluent at 1, 6, and 12 months after PD initiation. Flow cytometry was used to measure expression of IL-6R and Gp130 on isolated peritoneal leukocytes at the same time points. Peritonitis was an exclusion criterion. ♦ Results: At all time points, levels of IL-6 and sIL-6R, and the appearance rates of IL-6 (90.5 pg/min vs. 481.1 pg/min, p < 0.001; 138.6 pg/min vs. 1187.5 pg/min, p < 0.001; and 56.1 pg/min vs. 1386.0 pg/min, p < 0.001), sIL-6R (2035.3 pg/min vs. 4907.0 pg/min, p < 0.01; 1375.0 pg/min vs. 6348.4 pg/min, p < 0.01; and 1881.3 pg/min vs. 5437.8 pg/min, p < 0.01), and sGp130 (37.6 ng/min vs. 65.4 ng/min, p < 0.01; 39.2 ng/min vs. 80.6 ng/min, p < 0.01; 27.8 ng/min vs. 71.0 ng/min, p < 0.01) were significantly higher in peritoneal effluent from E-treated patients than from P-treated patients. Expression of IL6-R and Gp130 on individual leukocyte types isolated from PD effluent did not differ between E- and P-treated patients. The numbers of white blood cells present in effluent were higher in E-treated than in P-treated patients at all time points, but no significant differences were seen in the differential counts or in the number of exfoliated mesothelial cells. The IL-6 parameters in effluent from E-treated patients correlated with their plasma C-reactive protein. Despite the increased activation of the IL-6 system, no increase in peritoneal permeability as assessed by the dialysate-to-plasma ratio of creatinine in E effluent or by systemic inflammation was observed throughout the study. ♦ Conclusions: Higher levels of IL-6, its soluble receptors, and leukocyte expression were observed in E-treated than in P-treated patients, but this difference was not associated with alterations in peritoneal permeability or systemic inflammation during 1 year of follow-up. Leukocyte counts in effluent from E-treated patients were within the normal range previously reported for glucose solutions. This lack of clinical consequences may be a result of a parallel rise in sIL-6R and sGp130, which are known to control the biologic activity of IL-6. The utility of IL-6 level determinations, in isolation, for assessing the biocompatibility of PD solutions is questionable.

Unexpected inverse relationship between impaired glucose metabolism and lipoprotein-associated phospholipase A2 activity in patients with stable vascular disease

BACKGROUND Elevated lipoprotein-associated phospholipase A2 activity (aLp-PLA2) is associated with increased risk of cardiovascular events. In patients with stable atherovascular disease, we aimed to investigate whether impaired glucose metabolism might be associated with higher risk of elevated aLp-PLA2. METHODS We conducted a cross-sectional study in 825 stable patients after acute coronary syndrome, coronary revascularization or after first ischemic stroke (Czech part of EUROASPIRE III surveys). We measured aLp-PLA2 using diaDexus commercial kit. RESULTS In multiple step-wise regression analysis, the aLp-PLA2 was significantly positively associated with male gender, current smoking, LDL cholesterol and metabolic syndrome and negatively with statin treatment, body mass index and LDL/apoB ratio. After adjustment for these confounders, we observed an inverse relationship between aLp-PLA2 and fasting glycemia [β coefficient -2.18 (p<0.0001)] or glycated hemoglobin A1c (HbA1c) [β coefficient -5.89 (p<0.0001)]. Moreover, we found a positive association between aLp-PLA2 and pancreatic β cell function [β coefficient +0.10 (p<0.0001)], but not with an insulin sensitivity. CONCLUSION In present study, we cannot confirm any additive risk of impaired glucose metabolism in terms of increased activity of Lp-PLA2. On the contrary, presence of inadequately controlled diabetes mellitus was independently associated with lower risk of elevated aLp-PLA2 .

Asymmetric dimethylarginine, homocysteine and renal function – is there a relation?

Abstract The adverse effect of hyperhomocysteinemia on the vascular wall can be partially explained by increasing plasma concentration of asymmetric dimethylarginine (ADMA), a potent inhibitor of nitric oxide synthase. The aim of the study was to compare ADMA and homocysteine levels in three groups of subjects: blood donors with normal homocysteine concentration (group A), patients with hyperhomocysteinemia and normal kidney function (group B) and hemodialysis patients who are known to be hyperhomocysteinemic (group C). Concentrations of homocysteine (enzymatic method), ADMA (enzyme-linked immunoassay) and creatinine (Jaffe method) in EDTA plasma were measured. Plasma ADMA levels were significantly higher in both groups with hyperhomocysteinemia (1.60±0.56μmol/L in group B, 1.81±0.57μmol/L in group C) when compared with those in blood donors (0.82±0.29μmol/L, p<0.001 in both cases). Significant positive correlations were found between concentrations of ADMA and homocysteine (r=0.42, p<0.0001), ADMA and creatinine (r=0.39 p<0.001), homocysteine and creatinine (r=0.69, p<0.0001), age and homocysteine (r=0.47, p<0.001), age and ADMA (r=0.57, p<0.001) and age and creatinine (r=0.37, p<0.001). Increased ADMA concentrations in hyperhomocysteinemic patients were confirmed, but multiple linear regression analysis showed that this significant correlation is only apparent due the dependence of both parameters on age.