Lactate in cardiogenic shock - current understanding and clinical implications.

Adenosine triphosphate (ATP) is an essential substrate metabolite in human beings. Mitochondrial oxidative phosphorylation provides > 95% of ATP with the remainder derived from glycolysis or tricarboxylic acid cycle (TCA). In normal hearts, acetyl-CoA is synthesized from the ß-oxidation of free fatty acids (FFA) and the oxidation of pyruvate. Pyruvate is synthesized from glycolysis and can be submitted either for decarboxylation to acetyl-CoA or for dehydrogenation to lactate. Moreover, pyruvate, as well as lactate, plays a key role in aerobic glucose metabolism which is highly dependent on ubiquitous regulatory mechanisms. Many recent advances in molecular biology, genetics, and physiology have revealed new insights into the metabolic flux of lactate. The initial perception characterized by increased lactate production and accumulation in peripheral tissues in anaerobic conditions has been recently contested. The paradigm of increased lactate concentration in the anaerobic setting is discussed according to contemporary reports. Nevertheless, the clinical role of lactate as a prognostic factor in cardiovascular diseases is undisturbed, especially in the field of innovative technology of left/bi ventricular-assist devices and biochips where it reassured its diagnostic and prognostic impact on the cardiovascular system.

Prognosis of patients with implanted pacemakers in 4­year follow-up : Impact of right ventricular pacing site.

BACKGROUND: Pacing remains the method of choice for treatment of cardiac electrical conduction disorders. This study examined the interrelationship between the site of the right ventricular lead tip and patient prognosis in association with other cardiovascular risk factors over a 4­year follow-up period. PATIENTS AND METHODS: The study comprised 450 consecutive patients (223 women; aged 69.16 ± 9.63 years) who had their first SSI or DDD pacemaker implanted for typical indications. RESULTS: During follow-up, 91 (20.2%) patients died. The positive prognostic factors were: female sex (hazard ratio [HR] = 0.426), DDD pacemaker (HR = 0.526), oral anticoagulant use (HR = 0.330; all groups), sodium concentration (HR = 0.926), oral anticoagulant (HR = 0.115) and statin (HR = 0.260) use (female group), and non-apical location of the right ventricular lead tip (HR = 0.549; male group). Risk factors for death were: age (HR = 1.063), diabetes requiring insulin (HR = 2.832), creatinine concentration (HR = 1.005; all groups), age (HR = 1.11; female group), and elevated creatinine level (HR = 1.012; male group). In all patients, the non-apical location of the right ventricular lead tip was associated with an 18.92% reduced mortality rate during the 4­year follow-up, which was statistically significant for the male group. CONCLUSION: The non-apical location of the right ventricular lead tip was a positive prognostic factor and was statistically significant in the male subgroup.

One session of exercise regulates cathepsin B activity in the livers of trained and untrained rats.

Physical exercise causes adaptive changes, mainly in muscles, but it also influences other organs, including liver. Most changes are beneficial; however, strenuous exercise is a strong stressor, and it can result in splanchnic hypoperfusion with subsequent disturbances in liver homeostasis and energy. Cathepsin B is a protease linked to protein turnover and extracellular matrix degradation. It is also involved in autophagy and the activation of proinflammatory and profibrotic pathways. This study investigated the influences of one session of exercise and endurance training on the mRNA, protein level, and activity of cathepsin B in rat liver. Healthy rats were randomly divided into two groups (n = 30, each); one group was untrained and the other received 6-weeks of endurance training with an increasing load. For each group, rats were sacrificed before (controls, n = 10), immediately after (n = 10), and 3 h after (n = 10) an acute bout of intense exercise. Liver gene expression was evaluated with quantitative real-time PCR. Liver protein content was measured with ELISA. Liver enzyme activity was measured fluorometrically. One session of exercise or training did not influence cathepsin B gene expression or protein concentration at any investigated time point. In untrained rats, cathepsin B activity decreased 3 hours after (P = 0.027) one session of exercise. In trained rats, cathepsin B activity increased immediately (P = 0.005) after one session of exercise. Training did not influence baseline cathepsin B activity. In conclusion, one session of exercise differentially influenced cathepsin B activity in the liver, depending on training status.

Comparison of Two Methods for Determination of NGAL Levels in Urine: ELISA and CMIA.

BACKGROUND: Neutrophil gelatinase-associated lipocalin (NGAL) is a new useful biomarker for the early diagnosis of acute kidney injury. The aim of the study was to compare two analytical methods for measurement of urinary NGAL: enzyme-linked immunosorbent assay (ELISA) and chemiluminescent microparticle immunoassay (CMIA). METHODS: Two assays were used to measure urinary NGAL: ELISA kit (R&D Systems) and ARCHITECT Urine NGAL (Abbott Laboratories). The study material was the urine obtained from 30 healthy subjects (mean age 56.4 ± 15.2). RESULTS: The median value and interquantile range of urinary NGAL in the studied group measured by ELISA (R&D Systems) were 3.5 ng/ml (1.2; 6.6) and by CMIA (ARCHITECT Urine NGAL assay, Abbott Diagnostics) were 4.4 ng/ml (1.9; 9.4). Levels of urinary NGAL obtained by CMIA were significantly higher than by ELISA. There was a significant positive correlation between the concentration of urinary NGAL determined by both methods (r = 0.8625, P < 0.01). CONCLUSION: The comparison of individual data obtained by ELISA and CMIA should be taken with care. From laboratory's point of view, ELISA is less expensive than CMIA method for the determination of NGAL in urine. However, CMIA method allows rapid determination of urinary NGAL concentration through automated assay.

Strong association between fibronectin accumulation and lowered cathepsin B activity in glomeruli of diabetic rats.

In diabetic nephropathy the progressive accumulation of extracellular matrix (ECM) proteins results from an imbalance between synthetic and degradative pathways. While the role of the different matrix metalloproteinases in the impaired ECM degradation has been studied in detail, the function of lysosomal cysteine proteinases has not received adequate attention. The aim was to investigate a potential relationship between the accumulated ECM protein fibronectin (FN), and cathepsin B activity in isolated glomeruli of diabetic and healthy rats. Twenty male Wistar rats were included: 10 healthy and 10 with streptozotocin-induced diabetes. After 6 weeks, the experiments were terminated. In the homogenates of isolated glomeruli, FN content and cathepsin B activity were measured by ELISA or spectrofluorometry. FN was also analyzed by immunohistochemistry. Diabetic rats showed a significant rise of systolic blood pressure, impaired renal function and an enhanced urinary excretion of albumin, FN and cathepsin B. In the homogenates of the isolated glomeruli the ratios of FN/protein and FN/DNA showed a trend to higher values, while the ratios of cathepsin B/protein and cathepin B/DNA were reduced. The strong positive association between intraglomerular FN content and cathepsin B activity of in both groups suggests that this cysteine proteinase contributes to the degradation of the ECM protein FN. The much higher FN content in DN rats associated with an inadequate cathepsin B activity implies the role of an insufficient FN degradation by cathepsin B and other proteinases.

Inflammatory cytokines gene expression in bone tissue from patients with chronic rhinosinusitis- a preliminary study.

BACKGROUND: It is unclear whether mucosal inflammation has an effect on the bone under the mucosa in patients with chronic rhinosinusitis (CRS). OBJECTIVES: The aim of this study was evaluation of inflammatory cytokines genes expression in bone tissue taken from the patients who had undergone endoscopic sinus surgery for CRS. METHODS: A total group of a consecutive 49 patients with diagnosis of chronic rhinosinusitis based on EPOS 2007 criteria undergoing endoscopic sinus surgery for CRS were enrolled in the study. Based on histopathologic findings of the mucosal and bone tissues we evaluated the rate of inflammation. Expression of target genes: interleukin 1ß (IL1ß), interleukin 6 (IL6), interleukin 11 (IL11), tumor growth factor ß (TGF ß) and tumor necrosis factor α (TNF α) were analysed by real-time PCR method in samples of the ethmoid bone taken during endoscopic sinus surgery for CRS. RESULTS: Based on histopathological findings in the studied population we found symptoms of osteitis in 5 patients. In the studied population we found significant differences between patients with osteitis and without osteitis with respect to IL6 gene expression in bone tissue (p=0.0003), IL11 gene expression (p=0.02) and TNFα gene expression in bone tissue (p=0.0035). CONCLUSION: In our study we have demonstrated that in some patients with CRS and coexisting symptoms of osteitis some inflammatory markers genes expression are increased in this population.

Circulating electronegatively charged low-density lipoprotein in patients with angiographically documented coronary artery disease.

UNLABELLED: There is growing experimental evidence to suggest the role of oxidatively modified low-density lipoprotein (LDL) in the initiation and progression of atherosclerosis. The oxidation of lipoprotein moiety causes modification of positively charged lysine residues and results in negative net charge of lipoprotein particles. OBJECTIVE: To measure the amount of circulating electronegatively charged LDL particles (LDL-) in plasma of patients with angiographically documented coronary artery disease (CAD). METHODS: Thirty patients were assigned to the study group (CAD+) and 10 patients to the control group (Ctrl). LDL- was quantitated in homogeneous LDL fractions obtained by ultracentrifugation, using ion exchange high performance liquid chromatography. Plasma lipids were measured using enzymatic kits. RESULTS: The CAD+ group had significantly higher levels of LDL- in the whole LDL fraction (7.66+/-1.92 vs. 5.14+/-0.84%, p=0.0003). Moreover the CAD+ group had significantly higher levels of total cholesterol (255.4+/-35.1 vs. 210.4+/-22.4 mg/dL), LDL cholesterol (154.5+/-26.9 vs. 122.4+/-21.1 mg/dL) and significantly lower levels of high-density lipoprotein (HDL) cholesterol (40.4+/-9.4 vs. 51.0+/-11.5 mg/dL). LDL- remained significantly higher in the CAD+ group after adjustment for total cholesterol, LDL cholesterol and HDL cholesterol (6.3 vs. 5.14% at p=0.0095). There is a trend towards a positive correlation between LDL- levels and LDL cholesterol in the control group (Spearman R=0.55 at p=0.098). CONCLUSIONS: Electronegatively charged LDL appears to be an additional hallmark of coronary artery disease, independently of established lipid risk factors. The trend towards a positive correlation between LDL cholesterol concentration and the level of LDL- in the control group may reflect the susceptibility of LDL cholesterol to autoxidation, Moreover, this may indicate other oxidative mechanisms in coronary artery disease. Nonetheless, further studies assessing the prognostic value of electronegatively charged LDLs are necessary.

Selectin-P and interleukin-8 plasma levels in coronary heart disease patients.

BACKGROUND: Alterations of the immune system are now believed to play crucial role in the pathogenesis of atherosclerosis. The aim of this study was analysis of soluble forms of selectin-P and interleukin-8 levels in patients with different form of coronary heart disease. MATERIALS AND METHODS: In the study took part 18 patients with stable coronary heart disease, 20 patients with unstable coronary heart disease and 15 healthy persons from control group. Soluble selectin-P and interleukin-8 levels were measured in EDTA plasma with the use of enzyme immunoassay ELISA. RESULTS: The level of soluble selectin-P was significantly higher in unstable coronary heart disease patients in comparison to the stable coronary heart disease patients (P < or = 0.01) and nonsignificantly higher in comparison to the control group. The level of interleukin-8 were significantly higher in unstable coronary heart disease patients in comparison to the stable coronary heart disease patients (P < or = 0.01) and in comparison to the control group (P < or = 0.02). CONCLUSION: Our findings suggest that soluble form of selectin-P and interleukin-8 may be useful clinical predictors of unstable coronary heart disease. The assessment of the risk for the development of coronary heart disease requires further serial investigation.

Evidence of oxidative stress in the renal cortex of diabetic rats: favourable effect of vitamin E.

The aim of this study was to determine whether there are any disturbances of red/ox balance in the renal cortex of rats during the course of experimental diabetes. In the renal cortex of rats with streptozotocin-induced diabetes, the activity of superoxide dismutase (SOD) isoenzymes, glutathione peroxidase (GSH-Pox). glutathione S-transferase (GST) and glutathione reductase (GSH-RED) was measured in the 5th, 10th and 15th weeks of diabetes. Free radical cell damage was assessed on the basis of malonyldialdehyde (MDA) concentration. The influence of lipophilic antioxidant vitamin E on these analytes was also studied. An increase in MDA concentration in the 10th and 15th weeks of diabetes correlated significantly with plasma glucose concentration (r=0.47; p<0.001). Moreover, MDA concentration was influenced by time (+); p<0.001, diabetes (+); p<0.001, vitamin E (-) p<0.001 (ANOVA). Plasma creatinine concentration in rats was elevated by diabetes (p<0.001), whereas vitamin E decreased the concentration (p<0.05). Vitamin E lowered the activity of GSHPox (p<0.001) and GST (p<0.01) (ANOVA). Our results indicate that during experimental diabetes, disturbances of red/ox balance lead to disturbance in renal function manifested as increased creatinine blood concentration. We suggest that oral supplementation of vitamin E protects the renal cortex of rats during experimental diabetes.

Nephrotic origin hyperlipidemia, relative reduction of vitamin E level and subsequent oxidative stress may promote atherosclerosis.

BACKGROUND: The relation between nephrotic syndrome and atherosclerosis has not yet been fully clarified, although the high levels of low-density lipoprotein cholesterol usually found in this syndrome may give rise to atherosclerosis. This study was intended to test the disturbances of antioxidant/oxidant status in children with nephrotic syndrome (NS). METHODS: 8 children in the active stage (AS) of NS, 7 children during the remission stage (REM) of NS, and 14 control subjects (CTRL) were enrolled into the study. The levels of plasma total cholesterol (TC), HDL-cholesterol (HDL-chol), LDL-cholesterol (LDL-chol), triglycerides (TG), vitamin E and 7-ketocholesterol (7KCH) before and after plasma saponification were measured. RESULTS: A significant increase in the concentrations of TC, LDL-chol, vitamin E and total 7KCH in AS patients have been found. These patients had also a lower vitamin E/LDL-chol ratio. These changes have not been observed in the remission stage of nephrotic syndrome. Higher amounts of electronegatively charged-(oxidized) LDL particles as well as different oxysterols in AS patients have also been demonstrated. CONCLUSION: The study revealed significant disturbances in oxidant status during NS leading to plasma accumulation of oxidized LDL and cholesterol oxidation products that exert cytotoxicity and are known to induce atherosclerosis. We suggest that this may constitute an important link between nephrotic syndrome and atherosclerosis.

Impact of baseline level of 7-ketocholesterol on the rate of coronary reintervention in coronary artery disease patients--results of an observational study with questionnaire-based follow up.

INTRODUCTION: The aim of study was to assess the relation between plasma level of one of the oxysterols--7-ketocholesterol and angiographically evidenced coronary artery disease. MATERIAL AND METHODS: We measured plasma level of 7-ketocholesterol, malondialdehyde, total cholesterol, HDL cholesterol, triglycerides, erythrocyte activity of superoxide dismutase in 233 patients who previously underwent coronary angiography for diagnosis of coronary artery disease. The follow-up of patients (28 months) was done on basis of questionnaires sent out to them. RESULTS: We found that the subgroup of patients with 7-ketocholesterol above the median value (62 ng/mL) has higher rate of coronary reintervention (8/54 pts vs. 5/43 pts in 7-ketocholesterol below 62 ng/mL respectively, p = 0.284 Log-rank test). The finding was more pronounced, yet insignificant, in the subgroup of patients who underwent primary PTCA (6/19 pts vs. 2/20 pts, p = 0.1 Log-rank test). There were 3 patients in high 7-ketocholesterol subgroup who entered the study with previously undergone primary coronary intervention. There were not such patients in low 7-ketocholesterol subgroup. 4 deaths were observed in high 7-ketocholesterol subgroup, and one in low 7-ketocholesterol subgroup (they were predominantly not of cardiovascular origin). CONCLUSIONS: We failed to find a significant relation between 7-ketocholesterol and coronary artery disease, though there is a trend of such relation in patients who undergo PTCA, evidenced by a slightly higher number of repeated revascularizations among patients with high 7-ketocholesterol. The study indicates the areas of further studies with agents capable of lowering oxysterols and assessing also other end points.

[Troponin T--is it a marker of restenosis after transluminal percutaneous angioplasty in unstable angina patients?]. / Troponina T--marker restenozy po przezskórnej plastyce tetnic wiencowych u chorych na niestabilna chorobe wiencowa?

Troponin T (TpT) is a protein implicated in skeletal muscle contractions, including myocardium. It was shown that the presence of troponin TpT in unstable angina patients' blood is associated with poor prognosis. In the present study amongst 25 patients with unstable angina 12 were found to have TpT present in their blood. TpT concentration was higher in patients with III and IVo CCS symptoms in comparison with class I and IIo CCS symptoms: 0.207 +/- 0.275 and 0.144 +/- 0.186 ng/mL respectively (p = 0.053; nonparametric Kolmogorow-Smirnov test). Patients were subjected to percutaneous transluminal coronary angioplasty (PTCA). After 3 months of follow up 17 patients (the rest of them dropped out) were assigned to two groups: A (n = 8)--without and B (n = 9)--with clinical and electrocardiographic signs of restenosis. Retrospective analysis revealed the presence of TpT before PTCA in 6 group B patients and 2 group A patients. Relative risk of stenocardia recurrence was calculated as 2.25. TpT was present in the blood of 20 patients in the first 24 hours after PTCA, and group B patients had higher mean TpT concentration; that could result from reperfusion of more ischaemic myocardium. It seems that the presence of TpT in unstable angina patients' blood may be an important factor characterizing patients with more serious prognosis.

Origin of serum alpha-amylase isoenzymes from healthy adult rats.

In connection with common use of rats in experimental testing drug toxicity in the liver or pancreas, we checked the electrophoretic patterns of amylase isoenzymes extracted from the liver, pancreas, parotis and serum of healthy adult rats. Separation of isoenzymes was made by polyacrylamide gel electrophoresis. The results indicate the salivary glands and liver as sources of serum alpha-amylase.

Changes of the activities of urea cycle enzymes extracted from rat liver after surgical reduction of stomach capacity and after feeding a high-fat diet.

Fourty-two Wistar FL tribe rats were divided into control groups, fed a high-fat diet or subjected to surgical reduction of stomach capacity by 75%. After 60 days of experiment we observed decrease of animal body weight gain, as well as decrease of ornithine transcarbamylase and arginase activity. As indicated by the activity of these enzymes high-fat diet leads to an increase in urea cycle activity probably due to metabolic and hormonal imbalance. The utilisation of carbamoylphosphate for the pyrimidine biosynthesis pathway did not seem to change the ATC-ase activity.

[Measurements of amylase isoenzymes in sera and saliva of patients after radiotherapy because of larynx carcinoma]. / Aktywnosc izoenzymów amylazy sliny i osocza krwi pacjentów poddanych radioterapii z powodu nowotworów krtani.

Serum and salivary alpha-amylase were measured for controls and patients with laryngeal carcinoma before, and after localised irradiation including salivary glands. Additionally amylase isoenzymes in sera were measured using mini-column method. A significant increase in amylasemia was observed after irradiation. Alpha-amylase activity in saliva was decreased after irradiation but differences were not statistically significant due to the significant decrease of protein in saliva of irradiated group. An increase of salivary isoenzyme S activity was observed while pancreatic isoenzyme activity was not altered. This method allows easy differentiation of hyperamylasemia due to irradiation of parotid gland and disorders of the pancreas. Alpha-amylase activity measurements may detect metabolic changes in salivary glands after irradiation.