Diagnostic Accuracy of Brain-derived Neurotrophic Factor and Nitric Oxide in Patients with Schizophrenia: A pilot study.

BACKGROUND: Brain-derived neurotrophic factor (BDNF) and nitric oxide (NO) play multiple roles in the developing and adult CNS. Since BDNF and NO metabolisms are dysregulated in schizophrenia, we measured these markers simultaneously in the blood of schizophrenics and assessed their diagnostic accuracy. METHODS: Thirty-eight patients with schizophrenia classified according to demographic characteristics, symptomatologyand therapy and 39 age- and gender-matched healthy controls were enrolled. BDNF was determined by the ELISA technique while the concentration of nitrite/nitrate ([Formula: see text]) was measured by the colorimetric method. RESULTS: Serum BDNF levels were significantly lower (20.38±3.73 ng/mL, P = 1.339E-05), whilst plasma [Formula: see text] concentrations were significantly higher (84.3 (72-121) µmol/L, P=4.357E-08) in patients with schizophrenia than in healthy controls (25.65±4.32 ng/mL; 60.9 (50-76) µmol/L, respectively). The lowest value of BDNF (18.14±3.26 ng/mL) and the highest [Formula: see text] concentration (115.3 (80-138) µmol/L) were found in patients treated with second-generation antipsychotics (SGA). The patients diseased before the age of 24 and the patients suffering for up to one year had significantly lower serum BDNF levels than those diseased after the age of 24 and the patients who were ill longer than one year. Both BDNF and [Formula: see text] showed good diagnostic accuracy, but BDNF had better ROC curve characteristics, especially in patients with negative symptomatology. CONCLUSIONS: BDNF and nitrite/nitrate showed inverse changes in schizophrenic patients. The most pronounced changes were found in patients treated with second-generation antipsychotics. Although BDNF is not specific of schizophrenia, it may be a clinically useful biomarker for the diagnosis of patients expressing predominantly negative symptoms.

Ocular fundus abnormalities in patients with Balkan endemic nephropathy and other chronic kidney diseases.

AIM: The aim of this study was to examine the ocular fundus pathology in patients with Balkan endemic nephropathy (BN) and chronic kidney diseases (CKD). METHODS: The study included 51 patients with BN from the South Morava River region in Serbia, and 102 subjects with different stages of chronic renal diseases, matched according to age and gender, obtained from a database used in a recently published study. All patients had visited Outpatient Department of the Clinic of Nephrology, Clinical Center Nis. All patients underwent routine ophthalmic examinations. RESULTS: There were significantly more (P < 0.001) patients with age-related macular degeneration (AMD) in the group with BN (31.37 %) than in those with CKD (5.88 %). Multivariate logistic regression analysis confirmed that the significant factors related to AMD in the group with BN were albuminuria (P < 0.05) and proteinuria (P < 0.05); in CKD patients, the level of HDL (P < 0.05), while negative correlation with the level of triglyceride was registered (P < 0.05). There was no association between estimated glomerular filtration rate and AMD. The significant factors related to retinopathy in the group with BN are age (P < 0.05) and serum creatinine values (P < 0.05), in patients with CKD increasing age (P < 0.001) and DM (P < 0.05). CONCLUSION: Ocular fundus pathology in patients with BN is similar to the pathology of other CKD, but with significantly more AMD (about four times), probably related to the genetic/epigenetic factors.

Ocular and systemic factors associated with glaucoma in chronic kidney disease patients.

The goal of this study was to examine, the relationship between chronic kidney disease (CKD) and glaucomatous optic disc neuropathy in a cohort of patients from the south-east Serbia and to determine whether limited screening for glaucoma in specific subgroups of patients with CKD is reasonable and justifiable. This cross-sectional study included 328 subjects with various stages of CKD. All patients had visited the Outpatient Department of the Nephrology Clinic, Clinical Center Nis, Serbia. All patients underwent routine ophthalmic examinations. Glaucoma diagnosis based on elevated intraocular pressure (IOP), the presence of excavation of the optic nerve head (C/D ratio), and characteristic glaucomatous visual field loss (MD-mean deviation, PSD-pattern standard deviation). CKD was defined as kidney damage or glomerular filtration rate (GFR) of <60 ml/min/1.73 m(2) for >3 months. A total number of 328 CKD patients, 33 (10.1 %) with primary open angle glaucoma and 28 (8.5 %) with ocular hypertension (OH), were included in the study. Patients with CKD and glaucoma had significantly higher mean values of C/D ratio (0.59), visual field mean deviations (dB)-MD (p < 0.001), and visual field pattern standard deviations (dB)-PSD (p < 0.001) than patients with CKD and OH. Stepwise multivariate linear regression analysis confirmed that the most significant factors related to IOP are age (p < 0.05), AHT (p = 0.01), and eGFR (p = 0.001). Multivariate regression analysis also confirmed that the most significant factors related to cup-to-disc ratio are number of years of smoking (p < 0.05), AHT, and sCr (p < 0.01). In conclusion, the prevalence of glaucoma among CKD patients in the cohort from south-east Serbia is 10.1 %. Patients with CKD and glaucoma, eGFR and current cigarette smoking are associated with IOP level, MD, and PSD of visual field and C/D ratio.

Global and specific histone acetylation pattern in patients with Balkan endemic nephropathy, a worldwide disease.

UNLABELLED: Abstract Background: Balkan endemic nephropathy (BEN) is a chronic tubulointerstitial nephropathy present in the Danube river regions in several Balkan countries. There appears to be a polygenic susceptibility to the disease in interaction with multiple environmental factors (aristolochic acid, ochratoxin A). In a previous study SEC61G, IL17RA, HDAC11 proved to be differently methylated throughout all patient-control pairs of BEN patients from Serbia and Bulgaria. Emerging connections between DNA methylation and histone acetylation prompted the present study on histone acetylation in patients with BEN. METHODS: The study involved 39 patients with BEN, and 39 controls collected from non-endemic regions in Serbia. The EpiSeeker Histone H3 and H4 Total Acetylation Detection colorimetric Kits and specific acetylated at lysine 18 H3K18 and H3K36 acetylated at lysine 36 detection kits were used. RESULTS: It was documented that total H4 histone acetylation level was increased significantly, while total H3 histone acetylation did not differ significantly. Specific histone structure and functional properties may be affected by the observed derangement of H3 histone acetylation pattern, since H3K36 site was significantly more acetylated, while H3K18 tended to be less acetylated than in control subjects. Multiple regression analysis revealed a statistically significant relationship between H4, H3T and H3K36 in BEN patients. CONCLUSION: This preliminary study suggests that the acetylation of histone lysine residues was detectable and found increased at specific sites of H3 and total H4 histones isolated from urothelial cells of patients with BEN. Having in mind a possible mechanism and biological role of epigenetic chromatin modification in urothelial tumor development they obtained results may open opportunity for selective therapeutic interventions in patients with BEN.

Urinary protein patterns in patients with Balkan endemic nephropathy.

PURPOSE: Urinary excretion of beta2-microglobulin (beta2-MG), albumin, immunoglobulin G (IgG) and protein was examined in patients with Balkan endemic nephropathy (BEN), glomerulonephritis (GN) and healthy controls. METHODS: The proteins were measured in morning urine samples from 74 patients with BEN, 50 healthy persons and 22 patients with GN. RESULTS: In BEN patients, median values for albumin, beta2-MG and protein were above upper normal limits, but median IgG was inside normal range. All patients with GN had microalbuminuria (MAU) and half of them had increased urinary beta2-MG, which was also found in eleven patients with increased urinary IgG. In BEN patients, there were significant negative correlations between eGFR and all measured urinary proteins, the composition of which changed during the course of BEN. In patients with eGFR > 60 ml/min/1.73 m(2) isolated beta2-MG was the most frequent finding (10/12 patients), but MAU was present in 4/12 patients. In BEN patients with eGFR between 30 and 59 ml/min/1.73 m(2), beta2-MG appeared as often as the combination of beta2-MG and albumin and isolated MAU. Out of 49 BEN patients with eGFR > 30 ml/min/1.73 m(2) 15 had increased urinary IgG either alone (1) or together with beta2-MG (3) or albumin (3) or beta2-MG and albumin (8). In BEN patients with GFR < 30 ml/min/1.73 m(2) only 1/25 had isolated beta2-MG but increased urinary IgG with increased beta2-MG, and albumin was the most frequent. CONCLUSION: Although low-molecular weight proteinuria was the most frequent urinary finding in BEN patients, MAU was frequently detected in advanced stages of BEN but also in some patients with eGFR > 60 ml/min/1.73 m(2). IgG was increasingly found as eGFR decreased.

Toxic essential oils. Part II: chemical, toxicological, pharmacological and microbiological profiles of Artemisia annua L. volatiles.

Botanical drugs based on Artemisia annua L. (Asteraceae) are important in the treatment of malaria. Alongside with artemisinin, this aromatic species produces high and variable amounts of other chemicals that have mostly unknown biological/pharmacological activities. Herein, we have studied the toxicological/pharmacological profile of volatile constituents of a Serbian population of A. annua. Fifty-eight components were identified, among them, artemisia ketone (35.7%), α-pinene (16.5%) and 1,8-cineole (5.5%) were the most abundant ones. Significant variability of A. annua volatile profile was confirmed by means of agglomerative hierarchical cluster analysis indicating the existence of several different A. annua chemotypes. In an attempt to connect the chemical profile of A. annua oil with its biological/toxicological effects, we have evaluated in vivo and/or in vitro toxicity (including hepato- and nephrotoxicity/protection), antinociceptive, antioxidant (DPPH, ABTS and superoxide radical scavenging activity assays), enzyme inhibiting (protein kinase A and α-amylase) and antimicrobial potential of A. annua oil and of its constituents. Our results revealed that the beneficial properties of A. annua botanical drugs are not limited only to their antimalarial properties. Taking into account its relatively low toxicity, the usage of A. annua volatiles (at least of the herein studied population) does not represent a health risk.

The importance of L-arginine metabolism modulation in diabetic patients with distal symmetric polyneuropathy.

BACKGROUND: Micro and macrovascular disease is the most frequent cause of morbidity and mortality of the patients with diabetes mellitus. The recent investigations have pointed out the relationship between endothelial dysfunction and the progression of these diabetic complications, suggesting the crucial role of nitric oxide, vasodilator molecule of endothelial origin, in these events, including diabetic symmetric polyneuropathy. MATERIAL AND METHODS: The present study encompassed 100 individuals with diabetes mellitus type II and diabetic distal symmetric polyneuropathy (DSP). Nitrate+nitrite concentration, 3-nitrotyrosine, S-nitrosothiols, ADMA and SDMA levels and arginase activity were determined compared to the control group consisted of 50 age and sex matched voluntary blood donors. RESULTS: NO(2)+NO(3) concentrations, as well as 3-nitrotyrosine, S-nitrosothiol, ADMA and SDMA levels were significantly higher in patients with DSP compared to the control group. Plasma arginase activity in the patients with diabetic DSP was significantly lower compared to the values in plasma of control subjects. CONCLUSION: The obtained results confirmed that nitrate+nitrite, 3-nitrotyrosine, S-nitrosothiols, ADMA, SDMA and arginase activity determination in plasma of patients with diabetic DSP could be useful in monitoring the disease development and in assesing the therapy effects.

Schizophrenia is associated with increased levels of serum Fas and FasL.

BACKGROUND: A growing body of evidence suggests that the apoptotic process is dysregulated in schizophrenia. However, only a few studies have evaluated apoptotic markers in vivo in patients or their cell cultures. METHODS: Serum concentrations of Fas receptor (Fas/APO-1) and Fas ligand (FasL) were measured by ELISA techniques. The differences were tested according to the patients' demographic, clinical and drug treatment characteristics. The clinical accuracy of the examined markers was assessed using receiver operating characteristic (ROC) curve analysis. RESULTS: In this case-controlled study both sFas/APO-1 and FasL were significantly higher in the patients with schizophrenia than in the controls. An increase in apoptotic markers was independent of the symptomatology, drug treatment, heredity, the first onset of the disease, the duration of the psychotic disease as well as the tobacco abuse. A significant negative correlation between the duration of the disease and sFasL concentration was found. At the same time, a significant positive correlation was found between sFasL and lymphocyte caspase-3 activity. ROC curve analysis showed that sFasL was the most strongly associated with the presence of schizophrenia. CONCLUSIONS: We can conclude that the extrinsic apoptotic pathway is dysregulated in schizophrenia and sFasL may be a clinically useful disease predictor.

Transforming growth factor beta1, matrix-metalloproteinase-2 and its tissue inhibitor in patients with pseudoexfoliation glaucoma/syndrome.

BACKGROUND/AIM: Transforming growth factor-beta1 (TGF-beta1), oxidative stress and imbalance between matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) may play an important role in pathogenesis of pseudoexfoliation syndrome/glaucoma (PEX Sy/Gl). The aim of the study was to measure concentrations of TGF-beta1, MMP-2, TIMP-2 in the aqueous humor in the examined group, as well as to compare the biochemical findings with the following clinical parameters: degree of chamber angle pigmantation, presence of pseudoexfoliation and the value of intraocular pressure (IOP). METHODS: Aqueous samples from 30 patients with cataract, 30 patients with PEX Sy, 36 patients with PEX Gl, and 42 patients with primary open-angle glaucoma (POAG) were collected during phacoemulsification cataract surgery. TGF beta1, MMP-2, TIMP-2 Fluotokine Multi Analyze Profiling kits and Luminex technology were used to simultaneously measure TGF beta1, MMP-2 and TIMP-2. RESULTS: TGF-beta1, MMP-2, TIMP-2 were detected in human aqueous from all the groups with the highest level in the group with PEX Gl. Statistically, a significant correlation between the levels of TGF beta1, MMP-2, TIMP-2 in the aqueous humor of the patients with PEX Gl and the IOP value was confirmed (p < 0.05). In this group, the positive correlations between the TGF beta1 concentration in the aqueous humor and the presence of pseudoexfoliation (p < 0.01), on the one hand, and between the TIMP-2 level and the presence of pseudoexfoliation (p < 0.05), on the other, were reported. A statistically significant positive correlation of TGF-beta1 and MMP-2, and the degree of chamber angle pigmentation in the PEX Gl group was confirmed (p < 0.05). In the POAG group, TIMP-2 values were in a negative correlation with the degree of pigmentation (p < 0.05), and the IOP value (p < 0.05). CONCLUSION: TGF beta1 and MMP-2 affect the degree of chamber angle pigmentation and the degree of pseudoexfoliation in patients with pseudoexfoliative glaucoma.

Prognostic significance of microalbuminuria in patients with acute myocardial infarction.

BACKGROUND: Microalbuminuria has been reported to occur in patients with acute myocardial infarction (AMI) and associated with worse outcome. In our prospective analysis we included patients with AMI with the primary aim to examine whether urinary albumin excretion is increased in those patients and whether it is associated with worse in-hospital prognosis (major complications). The secondary objective was to examine the predictive power of microalbuminuria for 6-month mortality and re-hospitalization for cardiovascular disease. METHODS: One hundred thirty patients admitted to the Coronary Care Unit were studied prospectively. The diagnosis of myocardial infarction was based on the latest criteria of the European Cardiac Society. Microalbuminuria was defined as a urinary albumin creatinine ratio (UACR) and was measured on the third day after admission in the first morning urine sample. RESULTS: One hundred thirty patients were enrolled in this study--82 (63.03%) men and 48 (36.92%) women, age 62.48 +/- 12 years. A high proportion of study patients (27.7%) had microalbuminuria and 8.5% had overt albuminuria (UACR over 25 mg/mmol in men and over 35 mg/mmol in women) at the time of urine examination. During the hospital stay (average 7.6 +/- 3.0 days) 4 patients (3.1%) died from cardiovascular complications and all had microalbuminuria. In our study a high percentage of patients with in-hospital nonfatal complications had microalbuminuria but it did not have positive predictive association with the occurrence. During a 6-month follow-up period, 8 patients died from cardiovascular cause. In-hospital and total mortality (in-hospital and the during six-month follow-up) were significantly frequent in patients with microalbuminuria (p < 0.05). During a six-month follow-up period, 24 patients (18.5%) were re-hospitalized for cardiovascular disease and, among them, 54.2% had microalbuminuria. In univariant regression analysis microalbuminuria increased the risk for re-hospitalization, but multiple analysis didn't show the significance. CONCLUSIONS: We found that UACR measured during the first week after AMI is independently associated with increased long-term risk for in-hospital and six-month mortality. On the basis of these results, we suggest that this measurement should be included in the routine clinical work up of patients with AMI.

Lipoprotein(a) Is the Best Single Marker in Assessing Unstable Angina Pectoris.

This study evaluated whether statin therapy changed a diagnostic validity of lipid and inflammatory markers in ischemic heart disease (IHD) patients. Levels of lipids, lipoproteins, apolipoproteins, inflammatory markers, and atherogenic indexes were determined in 49 apparently healthy men and women, 82 patients having stable angina pectoris (SAP), 80 patients with unstable angina (USAP), and 106 patients with acute ST-elevation myocardial infarction (STEMI) treated or not treated with statins. Diagnostic accuracy of markers was determined by ROC curve analysis. Significantly lower apoA-I in all statin-treated groups and significantly higher apoB in statin-treated STEMI group compared to non-statin-treated groups were observed. CRP showed the best ROC characteristics in the assessment of STEMI patients. Lp(a) is better in the evaluation of SAP and USAP patients, considering that Lp(a) showed the highest area under the curve (AUC). Regarding atherogenic indexes, the highest AUC in SAP group was obtained for TG/apoB and in USAP and STEMI patients for TG/HDL-c. Statins lowered total cholesterol, LDL-c, and TG but fail to normalize apoA-I in patients with IHD.

Beta2-microglobulin and alpha1-microglobulin as markers of Balkan endemic nephropathy, a worldwide disease.

BACKGROUND: Urine beta2-microglobulin (beta2-MG) was mainly used as a tubular marker of Balkan endemic nephropathy (BEN) but recently alpha1-microglobulin (alpha1-MG) was proposed for the diagnosis of BEN. In this study, the potential of urine beta2-MG, alpha1-MG, albumin, and total protein in the differentiation of BEN from healthy persons and patients with glomerulonephritis (GN) and nephrosclerosis (NS) was examined. METHODS: This study involved 47 patients with BEN, 36 with GN, 11 with NS, 30 healthy subjects from BEN families, and 46 healthy subjects from non-BEN families. RESULTS: In BEN patients area under the curve (AUC) for urine beta2-MG (0.828) and alpha1-MG (0.782) was higher than for urine albumin (0.740), but in GN patients AUC for urine protein (0.854) and albumin (0.872) was significantly higher than for the two low molecular weight proteins. AUC for all four urinary markers in NS patients was significantly lower than in BEN patients, ranging between 500 and 595. Median urine beta2-MG excretion in BEN patients was 17.5 times higher than in GN patients and 18.3 times higher than in controls; median alpha1-MG excretion was higher only 3.0 and 2.25 times, respectively. In the differentiation of BEN from healthy controls, beta2-MG had higher sensitivity and specificity at the cutoff levels (p < 0.001) than alpha1-MG (p < 0.05). In the differentiation of BEN from GN, beta2-MG was the best marker. CONCLUSION: All four urinary markers can be used for the differential diagnosis of BEN, beta2-MG being the best. Like in aristolochic acid nephropathy, beta2-MG seems to be an early marker of tubular damage in BEN.

Serum Fas/FasL levels in dependence on clinical presentations of coronary disease and their relationship with risk factors.

BACKGROUND/AIM: Ischemic heart disease is mostly a consequence of atherosclerosis. Besides the inflammation, the Fas/Fas ligand (FasL)/caspase death pathway is documented to be activated in atherosclerotic lesions. The aim of this study was to compare the values of soluble forms of Fas and FasL in patients with different presentations of coronary disease and to correlate Fas/FasL with risk factors. METHODS: We studied 30 patients with chronic stable angina pectoris (SAP), 27 with non-stable angina pectoris (NSAP), and 39 with acute ST-elevation myocardial infarction (STEMI) and 27 age-matched healthy volunteers (the control group). Serum Fas/APO1 and FasL concentrations were determined using a commercially available enzyme-linked immunoassays (ELISA). RESULTS: Fas/APO-1 levels in the STEMI patients (6.981 +/- 2.689 ng/mL) were significantly higher than Fas levels in the controls (5.092 +/- 1.252 ng/mL, p < 0.01), but not significantly higher than Fas values in the SAP (5.952 +/- 2.069 ng/mL) and the USAP patients (5.627 +/- 2.270 ng/ml). Levels of FasL did not show any significant difference among the studied groups. In the SAP patients Fas/APO1 showed a significant positive correlation with high sensitivity C-reactive protein (hsCRP) (p < 0.05) and a negative correlation with high-density lipoprotein cholesterol (HDL-C) (p < 0.05), while FasL showed a significant positive correlation with low-density lipoprotein cholesterol (LDL-C) (p < 0.05). Fas levels between the patients having cholesterol within normal range and those whose cholesterol was above the normal range showed a significant difference (p < 0.05) only in the NSAP patients. Fas and FasL levels between the patients with hsCRP lower than 3.0 mg/L and those with hsCRP higher than 3.0 mg/L of the SAP group showed a significant differences (p < 0.001, p < 0.05, respectively). Strong correlation between Fas concentration and diabetes mellitus (p < 0.05) and FasL concentrations and both cholesterol (p < 0.01) and triglycerides (p < 0.01) in the NSAP patients was observed. The patients in the SAP group showed no strong correlation between Fas and FasL concentration and risk factors. CONCLUSIONS: The obtained results showed that apoptotic process is dysregulated in the patients with ischemic heart disease. Interdependence between Fas and FasL and inflammatory and lipid markers as well as with cardiovascular risk factors was established.

Plasma nitrite/nitrate concentrations in patients with schizophrenia.

BACKGROUND: Nitric oxide (NO) is known to be a signaling molecule with many physiogical functions including apoptotic process regulation. Since apoptosis may contribute to the pathophysiology of schizophrenia, this study was undertaken to determine the plasma concentrations of NO in schizophrenics. METHODS: Nitrite/nitrate (NO(2)(-)/NO(3)(-)) concentrations were measured in plasma from 40 patients with schizophrenia, and 36 age- and gender-matched healthy persons using a colorimetric test. RESULTS: Plasma NO(2)(-)/NO(3)(-) concentrations were significantly higher in patients with schizophrenia (102.8+/-34.7 micromol/L, p<0.0001) than in controls (69.2+/-13.2 micromol/L). Also, mean NO(2)(-)/NO(3)(-) values in female patients and controls were significantly higher (118.2+/-44.7 micromol/L, p<0.001; 74.8+/-16.1 micromol/L, p<0.05, respectively) compared to males (94.7+/-25.3 micromol/L, 67.6+/-10.8 micromol/L). Significant correlation was seen between plasma NO(2)(-)/NO(3)(-) concentrations and heredity, number of episodes and peripheral blood mononuclear cell (PBMC) caspase-3 activity, which was significantly higher in patients than in controls (p<0.05). There was no significant difference in NO(2)(-)/NO(3)(-) concentrations between patients with different Positive and Negative Syndrome Scale (PANSS) scores or between patients treated with haloperidol (97.2+/-31.2 micromol/L) and those treated with other atypical antipsychotic drugs (109.8+/-33.7 micromol/L). Both parameters showed no significant differences between smokers and non-smokers. CONCLUSIONS: This study showed that plasma NO(2)(-)/NO(3)(-) concentrations were significantly increased in patients with schizophrenia, being significantly higher in female than male patients, and showing a significant correlation with heredity, number of episodes and PBMC caspase-3 activity. These results suggest that NO could be considered an inducer or regulator of apoptosis in patients with schizophrenia.

Tubular marker excretion in children from families with Balkan nephropathy.

Balkan nephropathy (BN) has not been described in children; however, some previous studies in children from families with BN have revealed abnormalities of the urinary tract. In this study, urinary excretion of beta2-microglobulin, N-acetyl-beta-D-glucosaminidase (NAG) and gamma-glutamyl transpeptidase (GGT) was studied three times a year: spring, autumn, and winter, during a 3-year period, in 703 healthy children, initial age 9-13, from endemic and nonendemic settlements around the South Morava River. Beta-2-microglobulin excretion in urine, in all three seasons, was highest in children from families with BN compared with the excretion in children from the city, nonendemic villages, and those from nonendemic families. Increased urinary GGT excretion in children from endemic villages in October was higher than in children from the city and control villages, being the same in both endemic and nonendemic families. However, in February, it was similar in children from the city, endemic, and control villages. In conclusion, children from families with BN excreted significantly more beta2-microglobulin in all three seasons (spring, autumn, winter) of the study, in multivariate analysis significant for family status, gender, and the season (p < 0.001). NAG emerged as a potentially useful marker for seasonal exposure to an environmental nephrotoxin.

Polymorphisms of tumor-necrosis factor-alpha - 308 and lymphotoxin-alpha + 250: possible modulation of susceptibility to apoptosis in chronic lymphocytic leukemia and non-Hodgkin lymphoma mononuclear cells.

Tumor necrosis factor alpha (TNF-alpha) and lymphotoxin alpha (LT-alpha) have been shown to play an important role in the pathogenesis of limphoproliferative disease. Both cytokines regulate cell-survival and cell-death in leukemic cells. TNF-alpha and LT-alpha are highly produced in chronic lymphotic leukemia (CLL) and non-Hodgkin lymphoma (NHL) patients. Genetic polymorphism within regulatory regions of these cytokine genes can alter their expression levels. This study investigates an influence of TNF-alpha - 308 and LT-alpha + 250 polymorphisms on the activity of alkaline DNase in mononuclear cells of both patient groups as a potent biochemical marker of DNA fragmentation in the terminal phase of apoptosis. Study was performed on mononuclear cells of CLL and NHL patients. SNP were obtained by PCR-RFLP method. The activity of alkaline DNase was measured by spectrophotometric method. The study provided evidence of the influence of TNFG/A genotype and A alleles in the susceptibility to NHL, since the association of LT-alphaG/G genotype with CLL was observed. High-producing TNF-alpha - 308/LT-alpha + 250 heterozygous haplotype is associated with high NHL incidence. The investigated SNP influence the activity of alkaline DNase in CLL and NHL patients. The observed polymorphisms may modulate susceptibility of leukemic cells to apoptosis by way of DNase activity.

[Oxidative stress in human diseases].

Oxidative stress is a "privilege" of aerobic organisms. It can be induced by endogenous and exogenous factors. Most often, it is characterized by the production of free radicals and nonradical oxygen and nitrogen products, referred to under a single term "reactive species" (RS). Oxidative stress is a deleterious process that can be an important mediator of damage to cell structures, including lipids and membranes, proteins and DNA. However, reactive oxygen (ROS) and nitrogen species (RNS) are "two-faced" products. Produced in low/moderate concentrations as molecular signals that regulate a series of physiological processes, such as a defence against infectious agents, the maintenance of vascular tone, the control of ventilation and erythropoietin production, and signal transduction from membrane receptors in various physiological processes. Many of ROS-mediated responses protect cells against oxidative stress and maintain "redox homeostasis". Then, both reactive species are produced by strictly regulated enzymes, such as nitric oxide synthase (NOS), and isoforms of NADPH oxidase, or as by-products from not so well regulated sources, such as the mitochondrial electron-transport chain. An excessive increase in ROS production has been implicated in the pathogenesis of atherosclerosis, cardiovascular diseases, hypertension, ischemia/reperfusion injury, diabetes mellitus, neurodegenerative and immuno-inflammatory diseases. Within the cells, ROS can act as secondary messengers in intracellular signalling cascades, which can induce the oncogenic phenotype of cancer cells, cellular senescence and apoptosis.

Effects of regular exercise on cardiovascular risk factors profile and oxidative stress in obese type 2 diabetic patients in regard to SCORE risk.

AIM: The aim of the present study was to investigate not only the effects of aerobic exercise on overall cardiovascular risk factors profile and oxidative stress in obese, type 2 diabetic patients, but to elucidate if those effects depended on the previously estimated Systematic Coronary Risk Evaluation (SCORE) risk. SUBJECTS AND METHODS: Changes in several well-established cardiovascular risk factors and oxidative stress-defense parameters were measured in a total of 30 previously sedentary, obese type 2 diabetic patients, including 16 low-risk (SCORE < 5%, aged 48.8 +/- 6.0 years, with a mean BMI of 33.28 +/- 2.94 kg/m2) and 14 high-risk (SCORE > or = 5%, aged 56.3 +/- 6.9 years, with a mean BMI of 31.40 +/- 1.13 kg/m2) patients, in regard to the SCORE model, during six months of regular aerobic exercise, performed under supervision. RESULTS: Significant improvement was observed in the majority of cardiovascular risk factors, including body mass index, waist circumference, blood pressure, glycaemia, glycated haemoglobin, median blood glucose and lipid profile parameters in both diabetic subgroups during the exercise programme. However, the benefits of exercise on the majority of examined parameters became more evident in the low-risk subgroup, compared to the high-risk subgroup from baseline to 3 months. Regular exercise markedly reduced oxidative stress in both subgroups as well, as demonstrated for glutathione, plasma malondialdehyde, sulphydryl groups and catalase. CONCLUSION: Regular aerobic exercise, performed under supervision, has many beneficial effects in improving overall cardiovascular risk factors profile and reducing oxidative stress in both low-risk and high-risk (according to SCORE model), previously sedentary and obese type 2 diabetic patients.

Serum neopterin, nitric oxide, inducible nitric oxide synthase and tumor necrosis factor-alpha levels in patients with ischemic heart disease.

BACKGROUND: Atherosclerosis, a chronic inflammatory disease, underlies the pathogenesis of coronary artery disease. The present study assessed the diagnostic possibilities of inflammatory biomarkers, serum neopterin, nitrite/nitrate (NO2(-)/NO3(-)), inducible nitric oxide synthase (iNOS) and tumor necrosis factor-alpha (TNF-alpha), and their correlation with risk factors in patients with acute coronary syndromes and stable angina pectoris. METHODS: We studied 44 patients with chronic stable angina pectoris, 46 with unstable angina, 55 with acute ST-elevation myocardial infarction and 39 age-matched healthy volunteers (control group). Serum neopterin, iNOS and TNF-alpha were determined with commercially available enzyme linked immunosorbent assay methods and NO2(-)/NO3(-) by the modified cadmium-reduction method. RESULTS: Mean serum neopterin levels were significantly higher in patients with unstable and stable angina pectoris in comparison to control subjects (p<0.01 and p<0.05, respectively). Serum NO2(-)/NO3(-) values were significantly elevated (p<0.01) only in patients with unstable angina. ST-elevation myocardial infarction patients with cardiac death during follow-up showed significantly lower baseline neopterin values (p<0.001), and higher NO2(-)/NO3(-) levels (p<0.05) in comparison to those without adverse events. Significantly higher NO2(-)/NO3(-) values (p<0.05) were also found in patients who had myocardial reinfarction. Serum iNOS and TNF-alpha in all patient groups were within control ranges. A strong correlation was found between neopterin and both smoking (p<0.01) and triglycerides (p<0.05) in unstable angina patients. In stable angina patients, neopterin, iNOS and TNF-alpha significantly correlated with hypertension (p<0.01) and triglycerides (p<0.05). A significant difference in neopterin concentration was found between smokers and non-smokers (p<0.05). CONCLUSIONS: The results of this study suggest that in stable angina patients, if studied over time, serum neopterin or NO2(-)/NO3(-) levels may indicate future plaque instability. In ST-elevation myocardial infarction patients, neopterin and/or NO2(-)/NO3(-) levels may identify patients at long-term risk of death or recurrent acute coronary events after myocardial infarction.

Effects of aerobic exercise on microalbuminuria and enzymuria in type 2 diabetic patients.

Increased urinary albumin excretion is a strong predictor for the development of overt diabetic nephropathy and overall cardiovascular morbidity and mortality in patients with type 2 diabetes. In a previous study, regular aerobic physical activity in overweight/obese patients with type 2 diabetes mellitus was found to have significant beneficial effects on glycemic control, insulin resistance, cardiovascular risk factors, and oxidative stress. The aim of the present study was to investigate the effects of aerobic exercise in the same cohort of type 2 diabetic patients on urinary albumin excretion, serum levels and urinary excretion of enzymes, tubular damage, and metabolic control markers in type 2 diabetic patients. Changes from baseline to 3 and 6 months of aerobic exercise were assessed for urinary albumin excretion, serum activities, and urinary excretion of N-acetyl-beta-D-glucosaminidase (NAGA), plasma cell glycoprotein 1 (PC-1) and aminopeptidase N (APN), as well as their association with insulin resistance, cardiovascular risk factors, and oxidative stress parameters in 30 male type 2 diabetic patients (aged 54.8 +/- 7.3 years, with a mean BMI of 30.8 +/- 3.0 kg/m2). Microalbuminuria was found in six (20%) diabetic patients at baseline, three of them (10%) after three months, and only one patient (3.33%) at the end of the study period. A significant correlation was found for urinary albumin excretion at baseline both with sulfhydryl-groups and catalase, but not for urinary albumin excretion with MDA and glutathione. The prevalence of microalbuminuria tended to decrease after six months of aerobic exercise in type 2 diabetic patients, independently of any improvement in insulin resistance and oxidative stress parameters. Neither between-group nor within-group changes were found for urinary PC-1, APN, and NAGA activity. Serum NAGA was significantly increased (p < 0.05) over the control level in diabetic patients at baseline, but it decreased to the normal level after six months of exercise. This study has shown that a six-month aerobic exercise, without any change in the medication, tended to decrease microalbuminuria without changing enzymuria. However, further studies are needed not only to confirm those findings, but to elucidate potential mechanisms that would clarify the beneficial effects of exercise.