Association of polymorphisms in promoter region of TNF-α -238 and -308 with clinical outcomes in patients with immune-mediated inflammatory diseases on anti-TNF therapy.

The hypothesis of the study was that polymorphisms in promoter regions -238 and -308 of TNF-α could be associated with different clinical outcomes in inflammatory bowel diseases (IBD) and immune-mediated rheumatic diseases (IMRD). The aim was to examine the possible association of both polymorphisms with concentration of C-reactive protein (CRP) and fecal calprotectin (fCAL), onset of the remission and development of the ADA in patients on therapy with anti-TNF inhibitors. The prospective study was done in patients with IBD and IMRD on infliximab (IFX) or adalimumab (ADM). Patients were genotyped for TNF-α -238 and -308 polymorphisms. The concentration of CRP, fCAL, IFX or ADM and antibodies to drugs were measured according to manufacturer's instructions and followed-up for 6 or 12 months. Out of all patients (N = 112), number of patients in remission did not differ according to genotypes (for IBD patients P = 0.509 vs 0.223; for IMRD patients P = 0.541 vs 0.132 for TNF-α -238 and -308, respectively). Initial CRP concentration was higher in IBD patients with TNF-α -308 GG than GA/AA genotypes in patients who failed to achieve remission [11.8 (4.4-39.6) vs 3.1 (1.5-6.5), P = 0.033]. In IBD patients with remission, fCAL concentration after at least 6 months of therapy was higher in TNF-α-308 GG than in GA genotype [52 (25-552) vs 20 (20-20) µg/g, P = 0.041]. Our results showed the association of TNF-α -308 GG genotype with a higher concentration of CRP and fecal calprotectin in patients with inflammatory bowel diseases on IFX or ADM therapy. Clinical remission and development of antibodies to anti-TNF drugs were not associated with TNF-α -238 and -308 polymorphisms.

Lower concentration of vitamin D is associated with lower DAS28 and VAS-pain scores in patients with inflammatory rheumatic diseases treated with infliximab: a pilot study.

Vitamin D is beneficial in patients with immune-mediated rheumatic diseases as it has been shown that it lowers the incidence risk and the level of inflammation. To examine the association between clinical outcomes and initial 25-hydroxyvitamin D [25(OH)D] concentrations in patients with the immune-mediated rheumatic diseases treated with infliximab for 9 months. This study was performed in patients with rheumatoid arthritis (RA), ankylosing spondylitis (AS) and psoriatic arthritis (PsA) treated with infliximab for at least 38 weeks. Disease activity was assessed using Disease Activity Score (DAS28) for RA and PsA and Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) for AS, while the global assessment was performed using the Visual Analogue Scale (VAS). Patients were divided into 2 groups according to 25(OH)D concentration which was classified as deficient or non-deficient (below and above 50 nmol/L, respectively). Concentrations of infliximab (IFX) and C-reactive protein (CRP) were measured according to the manufacturer's instructions.This study was performed in patients with rheumatoid arthritis (RA), ankylosing spondylitis (AS) and psoriatic arthritis (PsA) treated with infliximab for at least 38 weeks. Disease activity was assessed using Disease Activity Score (DAS28) for RA and PsA and Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) for AS, while the global assessment was performed using the Visual Analogue Scale (VAS). Patients were divided into 2 groups according to 25(OH)D concentration which was classified as deficient or non-deficient (below and above 50 nmol/L, respectively). Concentrations of infliximab (IFX) and C-reactive protein (CRP) were measured according to the manufacturer's instructions. The study included 23 patients (14 with RA, 6 with AS and 3 with PsA), median age 54 years, 15 females. Vitamin D deficient and non-deficient groups had median initial concentrations of 38 and 61 nmol/L, respectively. DAS28 and pain on VAS calculated at the 2nd and 38th week showed a statistically significant decrease only in RA and PsA patients with vitamin D deficiency (P = 0.02 and 0.06, respectively). Lower initial concentration of 25(OH)D in patients treated with infliximab was associated with better improvement of clinical measures (DAS28 and VAS) of disease after 9 months of therapy.

Effect of extract from Boswellia serrata gum resin on decrease of GAD65 autoantibodies in a patient with Latent Autoimmune Diabetes in Adults.

BACKGROUND: Extracts from Boswellia serrata gum resin have anti-inflammatory effect and are used for treatment of a variety of chronic inflammatory diseases. It was previously demonstrated that the treatment with Boswellia serrata gum resin of LADA (Latent Autoimmune Diabetes in Adults) patients decreased blood levels of IA2 antibodies, one of the markers associated with LADA autoimmune diabetes. PRIMARY STUDY OBJECTIVE: The purpose of this study was to test whether Boswellia serrata gum resin also influences GAD65 autoantibodies as the other marker associated with LADA. METHODS/DESIGN: We report a case study of male patient diagnosed with LADA with positive GAD65 autoantibodies who was treated with extract from Boswellia serrata gum resin, during 9 months. Blood levels of GAD65 autoantibodies, fasting blood glucose levels and HbA1c were measured before the treatment and periodically during the treatment. RESULTS: Over the observed period, the blood levels of GAD65 autoantibodies linearly decreased about 25%. CONCLUSION: The study confirms that extract of Boswellia serrata gum resin seems to prevent insulitis in patients with LADA, as indicated by its action on both markers of autoimmune diabetes, i.e., GAD65 and IA2 autoantibodies. The possibility that the treatment with boswellic acids of LADA patients with positive autoantibodies could be beneficial on the course of the disease, calls for further investigation and a clinical study.

CHOLESTEROL AND CHLORIDE IN ACUTE HEART FAILURE.

Detecting predictors of poor outcome is crucial for understanding the underlying pathophysiology of heart failure (HF) and thus creating new therapeutic concepts. It is well established that low serum lipid levels are associated with unfavorable outcomes in HF patients. Several studies examined the association between serum lipids and established predictors of mortality in HF patients. The aim of the present study was to examine the association of serum lipid and chloride concentrations, as well as their impact on survival in acute heart failure (AHF). The present study was performed as a prospective, single-centre, observational research. The study included 152 patients with AHF. Spearman's correlation coefficient revealed a significant positive correlation of serum chloride levels with serum levels of total cholesterol (ρ 0.221, p=0.006), low-density lipoprotein cholesterol (LDL-c) (ρ 0.187, p=0.015) and high-density lipoprotein-cholesterol (HDL-c) (ρ 0.169, p=0.038). Binary logistic regression revealed a significant association of chloride, total cholesterol and LDL-c serum levels measured at admission with hospital survival (OR 1.077, CI 1.01-1.154, p=0.034), (OR 1.731, CI 1.090-2.748, p=0.020) and (OR 1.839, CI 1.033-3.274, p=0.038), respectively, as well as with 3-month survival (OR 1.065, CI 1.002-1.131, p=0.042), (OR 1.625, CI 1.147-2.303, p=0.006) and (OR 1.711, CI 1.117-2.622, p=0.014), respectively. In conclusion, positive statistical association between serum cholesterol (total cholesterol, LDL-c and HDL-c) and chloride levels may suggest their similar modulation by AHF pathophysiology. Serum levels of total cholesterol, LDL-c and chloride contribute to patient survival.

Ascorbic acid-A black hole of urine chemistry screening.

BACKGROUND: Study was performed in order: (i) to assess the comparability of glucose, bilirubin, hemoglobin, leukocyte esterase, and protein; (ii) to assess accuracy of glucose, bilirubin, hemoglobin, leukocyte esterase, and protein; and (iii) to evaluate interference of ascorbic acid on the glucose, bilirubin, hemoglobin, and nitrite determination using 2 different dipsticks: iChem Velocity, Iris Diagnostics and Combur-10M, Roche Diagnostics. METHODS: Random urine specimens were included in the study. Comparability, accuracy, and ascorbic acid interference testing were performed. RESULTS: Obtained results have shown almost perfect agreement for all parameters between 2 dipsticks in samples with negative ascorbic acid. Agreement in samples with positive ascorbic acid was not acceptable for bilirubin, protein, nitrite, and hemoglobin. Accuracy was not acceptable for hemoglobin and leukocyte esterase on both dipsticks. Ascorbic acid interference examination has shown that intensity of interference differs between dipsticks. Ascorbic acid interferes with glucose, hemoglobin, nitrite, and bilirubin at different concentrations causing false-negative results. CONCLUSION: Obtained results indicate that it is necessary to determine diagnostic accuracy of used dipstick in order to define purpose of urinalysis. It is very important to choose dipstick with ascorbic acid indicator and to examine ascorbic acid impact on dipstick analytes independently of manufacturer claims.

Kappa Free Light Chains in Cerebrospinal Fluid of Patients with Identified Oligoclonal Immunoglobulin G.

BACKGROUND: Production of kappa free light chains (KFLC) represents a part of humoral immune response, along with the synthesis of intrathecal immunoglobulins. Increased concentrations of immunoglobulin G light chains, kappa and lambda chains, were identified through research of numerous diseases of central nervous system. The qualitative method of isoelectric focusing (IEF) followed by immunofixation currently represents the accepted standard in identifying oligoclonal bands (OCB), but establishing a sensitive immunonephelometric method for quantification of kappa free light chains (KFLC) in cerebrospinal fluid (CSF) has paved a way for new diagnostic possibilities. Andersson classified the pattern types of OCB, ranging from type 1 to type 5, wherein types 2 and 3 indicate intrathecal synthesis. Our aim was to determine KFLC in CSF of patients with clinically isolated syndrome (CIS) who had presented with type 2 and type 3 OCB, to determine if there is a difference in concentrations between those two groups and to establish a borderline value of KFLC which would enable differential diagnostics. SUBJECTS AND METHODS: 70 patients, who underwent lumbar punction for CSF analysis and had their blood sampled through the cubital vein, participated in the study. Patients were classified according to Andersson as type 2 or type 3, which besides adulthood, represented the inclusion criteria. The average age of patients classified as type 2 was 36 years, and those classified as type 3 was 39 years, where it is evident that there was not a statistically significant difference (p=0.0685). We used a qualitative electrophoretic technique of IEF with agarose gel followed by immunofixation, and a quantitative immunonephelometric method. All results were interpreted on a level of statistic significance of p<0.05. RESULTS: CSF KFLC concentrations in type 3 were statistically and significantly elevated with regard to type 2 (Mann-Whitney test, p=0.0430). The median for KFLC in type 2 was 0.9 mg/L, while the median for KFLC in type 3 was 2.71 mg/L, and the detection limit for both types was 0.18 mg/L. We used a statistical ROC curve to determine that KFLC concentration can be used for differential diagnostics, meaning it can discriminate type 2 from type 3 with clinical sensitivity of 61% and clinical specificity of 71% (AUC=0.641) (p=0.037). CONCLUSION: Despite the obtained statistically significant differences in concentrations of KFLC between types of OCBs and ROC analysis results, determination of KFLC by a nephelometric method, insufficiently strong clinical sensitivity and specificity does not justify abandonment of IEF method followed by immunofixation.

Increased calcium-independent lipoprotein phospholipase A2 but not protein S100 in patients with schizophrenia.

BACKGROUND: The aim of this paper was to investigate serum concentrations of calcium-independent lipoprotein phospholipase A2 (PLA2) and protein S100 in schizophrenia patients in comparison to healthy controls and correlate them with the clinical severity, duration, and number of schizophrenia relapses. SUBJECTS AND METHODS: This study included 65 schizophrenia patients and 70 controls. Schizophrenia was diagnosed according to DSM-IV-TR criteria. Clinical severity was determined by PANSS. PLA2 and protein S100 concentration were assessed by the enzyme-linked immunosorbent assay (ELISA). RESULTS: PLA2 concentrations were higher in patients with schizophrenia, whereas protein S100 concentrations were not. Higher concentrations of PLA2 were positively correlated with the duration of illness and number of episodes, as determined by multivariate analysis. CONCLUSION: PLA2 might be considered a possible biochemical trait marker for schizophrenia. Further research with larger and more homogeneous clinical samples is required.

Effect of Extra Virgin Olive Oil on Biomarkers of Inflammation in HIV-Infected Patients: A Randomized, Crossover, Controlled Clinical Trial.

BACKGROUND: Premature atherosclerosis in HIV-infected patients is associated with chronic infection by itself and adverse effects of antiretroviral treatment (ART). Extra virgin olive oil (EVOO) has a beneficial effect on the cardiovascular system because of its anti-inflammatory properties. The objective of this study was to determine whether the consumption of EVOO improves inflammation and atherosclerosis biomarkers in HIV-infected patients receiving ART. MATERIAL AND METHODS: This randomized, crossover, controlled trial included 39 HIV-positive male participants who consumed 50 mL of EVOO or refined olive oil (ROO) daily. Four participants dropped out of the study. Leukocyte count, erythrocyte sedimentation rate (ESR), high-sensitivity C-reactive protein (hsCRP), interleukin-6, fibrinogen, total cholesterol, LDL cholesterol, HDL cholesterol, triglycerides, malondialdehyde, glutathione-peroxidase, superoxide dismutase, oxidized LDL and von Willebrand factor were determined before the first and after each of the 2 intervention periods. Intervention and washout periods lasted for 20 and 14 days, respectively. RESULTS: In participants with >90% compliance (N=30), hsCRP concentrations were lower after EVOO intervention (geometric mean [GM], 1.70 mg/L; 95% confidence interval [CI], 1.15-2.52) compared to ROO administration (GM, 2.92 mg/L; 95% CI, 1.95-4.37) (p=0.035). In participants using lopinavir/ritonavir, ESR and hsCRP concentrations decreased 62% and 151%, respectively, after EVOO administration. In the whole study population (N=35) we found no difference in analyzed biomarkers after EVOO administration. CONCLUSIONS: Our exploratory study suggests that EVOO consumption could lower hsCRP in patients on ART.

[Compensated creatinine method and glomerular filtration rate estimation in a heterogeneous population of patients]. / Kompenzirana metoda za odredivanje kreatinina i procjena glomerularne filtracije u heterogenoj populaciji bolesnika.

Creatinine is a metabolite excreted mainly by glomerular filtration, which makes it an important endogenous indicator of kidney function. Creatinine clearance is defined as the ratio of the concentration of creatinine in serum and urine. It assesses glomerular filtration. Creatinine and creatinine clearance have the leading role in the early diagnosis, monitoring and classification of chronic kidney disease. The routine method for determining the concentration of creatinine is the Jaffé photometric method. A newer version is the compensated method. Furthermore, the recommended equation for the estimation of glomerular filtration rate (GFR) is the one based on the MDRD study (eGFR) intended for people over 18 years. The aim of the study was to evaluate how the introduction of the compensated method would affect the clinical use and influence the assessment of GFR in the interpretation of findings and treatment monitoring for people over 20 years. The study group included 130 men and 142 women whose requested laboratory test was creatinine clearance. Data were collected over 20 days at Sestre milosrdnice University Hospital. Serum creatinine concentration and eGFR were determined by the compensated and uncompensated Jaffé method. In conclusion, the compensated creatinine method is not statistically comparable with the uncompensated method, but is clinically fully applicable to the general population above the age of 20, given that the reference intervals are changed. Comparison of eGFR as estimated by the compensated and uncompensated methods to determine creatinine concentration showed the same results as the comparison of clearance. Using the compensated method yielded statistically incomparable results in GFR estimation. However, in clinical practice, patient classification according to stages of chronic kidney disease (CKD) was comparable in the male group according to clearance and eGFR (pi=0.922 and pi=0.230, respectively), while the female group was classified significantly different according to clearance and eGFR (pi<0.016 and pi<0.001, respectively). Switching to the compensated creatinine method while simultaneously applying the eGFR formula was shown to be valid, as patient classification according to CKD stages was comparable (pi=0.921); thus, the methods are reliable for use instead of creatinine clearance in the general population with various diagnoses, which can be noted in all laboratories and which is, although inhomogeneous, routinely used to measure daily creatinine clearance.

Differential diagnostic performance of Rose Bengal Score Test in Sjøgren's syndrome patients.

Aim of the study was to evaluate the diagnostic performance of the Rose Bengal score test for Sjøgren's syndrome (SS), and to explore differences between other tests and examinations. All participants were examined, including (but not limited to) unstimulated (UWS) and stimulated (SWS) whole saliva, labial gland biopsy (LGB or focus score), ophthalmologic questionnaire (ocular surface disease index OSDI) and objective tests: Schirmer test 1 (Sch. 1), Schirmer test 2 (Sch.2), Tear Break-up Time (TBUT) test and Rose Bengal score (RBS). Data were analyzed using Mann Whitney U-test, Receiver Operating Characteristic analysis, with specificity and sensitivity calculations and Spearman's correlation test. ROC curves showed a poor diagnostic performance of TBUT and OSDI. Sch. 1, Sch.2 and LGB all exhibited a high diagnostic performance. RBS exhibited the best performance (sensitivity 100,00; specificity 100,00; AUC 1,000). Study reveals the scarce reliability of TBUT, OSDI and Sch.1, and emphasizes RBS as the test of choice in the SS diagnosis.

Serum Brain-derived neurotrophic factor (BDNF): the severity and symptomatic dimensions of depression.

INTRODUCTION: The aim of this study was to compare the concentration of serum Brain-derived neurotrophic factor (BDNF) in patients suffering from major depressive disorder (MDD) considering the severity of MDD episode defined by the Hamilton rating scale for depression (HAMD-17). The other aim was to research the connection between serum BDNF and the symptomatic dimensions of MDD. SUBJECTS AND METHODS: The study includes 139 participants with major depressive disorder (MDD). Diagnosis of MDD was set by DSM-IV-TR criteria. The severity of MDD was estimated with HAM-D-17 in the manner that mild episode was diagnosed if the score on HAMD-17 was up to 18, moderately severe 18-25 and severe over 25. Concentration of BDNF was determined by the ELISA method. RESULTS: This research could not find a difference in BDNF concentration considering the severity of the depressive disorder in groups suffering from mild, moderately severe and severe episodes of MDD (F=1.816; p=0.169). Factor analysis of HAMD-17 extracted four dimensions of depressive symptoms. None of the symptomatic dimensions was significantly related to BDNF concentration. CONCLUSION: Results of this study indicate that serum BDNF levels are not related to the severity of depression and its specific symptomatic dimensions. These findings support the idea of a complex relationship between BDNF concentration at the periphery and in the CNS.

The prevalence of preanalytical errors in a Croatian ISO 15189 accredited laboratory.

BACKGROUND: The preanalytical phase is the most common source of laboratory errors. The goal of this descriptive study was to analyze the prevalence and type of preanalytical errors in relation to the site of sample collection (inpatient vs. outpatient) and the type of laboratory unit (hematology and coagulation vs. biochemistry). For the biochemistry unit, the data were also analyzed relative to the type of the analysis (stat vs. routine). METHODS: We retrospectively analyzed the sample and test request form error rate for a 1-year period, from January to December 2008. RESULTS: The frequency of the sample errors differed significantly between the emergency and routine biochemistry unit (0.69% vs. 2.14%; p<0.0001), and between inpatients and outpatients (1.12% vs. 1.36%; p=0.0006). Hemolysis was the most frequent sample error, accounting for 65% of all unsuitable specimens in the emergency biochemistry unit. The total sample error rate did not differ between hematology and coagulation vs. the biochemistry unit. The frequency of test request form errors differed significantly with respect to the sample collection site (p<0.0001), laboratory unit (p<0.0001) and type of the analysis (p<0.0001). Errors in the test request form were least frequent in the outpatient unit (2.98%) and most frequent in the routine biochemistry unit (65.94%). CONCLUSIONS: Sample and test request form errors in our laboratory are occurring with a frequency comparable to that reported by others. Continuous educational action is needed for all stakeholders involved in laboratory testing to improve the quality of the preanalytical phase of the total testing process.

Antibodies to mutated citrullinated vimentin and antibodies to cyclic citrullinated peptides in juvenile idiopathic arthritis.

BACKGROUND: The goal of the study was to assess the presence of antibodies to mutated citrullinated vimentin (anti-MCV) and cyclic citrullinated peptides (anti-CCP) in patients with juvenile idiopathic arthritis (JIA) compared with patients with other juvenile onset rheumatic diseases. METHODS: The study included 56 patients who fulfilled the International League of Associations for Rheumatology (ILAR) classification criteria for JIA, and 17 control patients with other juvenile onset rheumatic diseases. Data on six core outcome variables and the Sharp score were collected for patients with JIA. Sera and synovial fluid, if available, were tested for anti-CCP and anti-MCV antibodies using a commercial enzyme-linked immunosorbent assay (ELISA). RESULTS: Anti-MCV antibodies were positive in 3/56 (5.4%) and anti-CCP in 1/56 (1.8%) of patients with JIA. Two out of three anti-MCV positive patients (one of them also anti-CCP positive) were found to be rheumatoid factor (RF)-positive with polyarticular disease. Within the control group, anti-MCV was positive in 4/17 (23.5%) patients, while anti-CCP positivity was not observed. No correlation between anti-MCV with anti-CCP antibody levels was found for any of the six core outcome variables or for the adapted Sharp score. CONCLUSIONS: Our results show that antibodies targeting citrullinated proteins are not a useful diagnostic marker for JIA, but can indicate severe patterns of disease in JIA.

Relevance of KFLC quantification to differentiate clinically isolated syndrome from multiple sclerosis at clinical onset.

OBJECTIVE: The objective of the research was to evaluate diagnostic and predictive value for determination of KFLC in cerebrospinal fluid (CSF) compared to the qualitative procedure of OCB determination in patients with CIS who converted to MS during a two-year period. PATIENTS AND METHODS: KFLC, total immunoglobulin G (IgG), serum albumin and CSF albumin were determined with an immunonephelometric method in 151 patients with suspected MS who were admitted to the Neurology Clinic while CSF/serum quotients (QKFLC, QIgG and QAlb) and indexes were calculated with regards to albumin (QCSF/Qserum). Presence of OCBs was determined by isoelectric focusing with immunofixation. Based on their clinical, OCB and magnetic resonance imaging (MRI) findings, 50 patients were classified as other neurological disorder patients (OND), and 101 patients were classified as CIS, 50 of which converted to MS during the two-year period. ROC analysis, ROC curve comparisons and comparison of median KFLC parameters were used to find optimal cut-off with regards to CIS diagnosis and conversion to MS. RESULTS: CSF KFLC median was 2,01 mg/L in MS group contrary to 0,68 mg/L and 0,17 mg/L in CIS and OND group, and KFLC index was 33,52 mg/L contrary to 9,68 mg/L and 3,71 mg/L (p < 0,0001). ROC analysis for accuracy of detection of intrathecal synthesis for QKFLC and KFLC index showed an AUC of 0,891 and 0,839 and the cut-off of 0,027 and 8,82, respectively (sensitivity 73,2% and 71,3%; specificity 96,0% and 98,0%; +PV 97,4% and 98,6%). The diagnostic accuracy of KFLC index for conversion from CIS to MS showed AUC of 0,840 and a cut-off of 9,092 (sensitivity 90,0%; specificity 73,3%; -PV 93,7%). Life age correlates significantly with serum KFLC (r = 0,34; p < 0,0001) and through aging process lower KFLC indexes can be expected, i.e. likelihood of false negative diagnoses. CONCLUSION: KFLC index showed diagnostic value, although it is not more specific and more sensitive than OCB. Application of KFLC might serve as a screening method while OCB could be used in uninterpretted cases only. Patients who converted to MS have significantly higher KFLC which can contribute to an early diagnosis and prompt therapy with its predictive role.

Comparison of Enzymatic Assay for HBA1C Measurement (Abbott Architect) With Capillary Electrophoresis (Sebia Minicap Flex Piercing Analyser).

OBJECTIVE: To compare the analytical performances of the enzymatic method (EM) and capillary electrophoresis (CE) for hemoglobin A1c (HbA1c) measurement. METHODS: Imprecision, carryover, stability, linearity, method comparison, and interferences were evaluated for HbA1c via EM (Abbott Laboratories, Inc) and CE (Sebia). RESULTS: Both methods have shown overall within-laboratory imprecision of less than 3% for International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) units (<2% National Glycohemoglobin Standardization Program [NGSP] units). Carryover effects were within acceptable criteria. The linearity of both methods has proven to be excellent (R2 = 0.999). Significant proportional and constant difference were found for EM, compared with CE, but were not clinically relevant (<5 mmol/mol; NGSP <0.5%). At the clinically relevant HbA1c concentration, stability observed with both methods was acceptable (bias, <3%). Triglyceride levels of 8.11 mmol per L or greater showed to interfere with EM and fetal hemoglobin (HbF) of 10.6% or greater with CE. CONCLUSION: The enzymatic method proved to be comparable to the CE method in analytical performances; however, certain interferences can influence the measurements of each method.

Association of methylenetetrahydrofolate (MTHFR) and apolipoprotein E (apo E) genotypes with homocysteine, vitamin and lipid levels in carotid stenosis.

The aim of the study was to investigate the association between methylenetetrahydrofolate (MTHFR) genotypes and levels of homocysteine (Hcy), folate, vitamin B12 and lipids as well as the association between apolipoprotein E (apo E) genotypes and levels of lipids in a Croatian healthy control group and a group of patients with > 70% carotid stenosis (CS). The study included 98 Croats, 38 patients with > 70% carotid stenosis and 60 age- and sex-matched controls. The MTHFR and apo E genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), Hcy by enzyme immunoassay, vitamins by immunochemiluminiscence, and lipids by spectrophotometric method. There was no difference between control subjects and CS patients in the distribution of C677T MTHFR genotypes (p=0. 786) and alleles (p=0.904), however, differences in the frequencies of apo E genotypes (p=0.012) and alleles (p=0.029) were statistically significant. The odds ratio for apo E 3/4 genotype was 3.93 (95% CI 1.23-12.61). Hyperhomocysteinemia (> or =15 micromol/L) was found in 11% of CS patients and 5% of control subjects. Total cholesterol, triglycerides, vitamin B12 and folate were statistically different in "all MTHFR genotypes" (p<0.001, p<0.01, p=0.044 and p=0.036, respectively), and in TC/TT (p<0.001, p=0.003, p=0.030 and p=0.032, respectively) groups. The levels of total cholesterol, LDL cholesterol and triglycerides in the apo E 3/3, and total cholesterol in the apo E 3/4 group yielded statistical difference. An association was found of apo E 3/4 genotype but not of MTHFR genotypes with the risk of CS. MTHFR and apo E affect blood lipid levels, which was statistically confirmed. An association was also recorded between hyperhomocysteinemia and patients with CS. Vitamin status in CS showed a statistically verified association with TC/TT MTHFR genotype. In the group of patients with TC/TT MTHFR genotype, lower vitamin B12 and higher folate values were recorded. The results of multiple logistic analysis showed that there was no statistical significance of Hcy levels (OR 2.403, p=0.334) or conventional vascular risk factors such as smoking habit (OR 0.505, p=0.149), age (OR 1.048, p=0.087) or sex (OR 2.037, p=0.112) in predicting CS.

Hypochloraemia as a predictor of developing hyponatraemia and poor outcome in acute heart failure patients.

AIMS: Heart failure (HF) is a major public health issue currently affecting more than 23 million patients worldwide. Hyponatraemia has been shown to be a predictor of poor outcome in patients with acute and chronic HF. Therefore, we aimed at finding a marker for early detection of patients at risk for developing hyponatraemia. To this end, the present study investigated the relationship between initial serum chloride and follow-up sodium levels in acute heart failure (AHF) patients. METHODS AND RESULTS: The present study was performed as a prospective, single-centre, observational research with a total of 152 hospitalised AHF patients. Compared to patients with initial normochloraemia, patients with initial hypochloraemia had a statistically significantly higher incidence of hyponatraemia after a 3-month follow-up [P<0.001; odds ratio (OR)=27.08, CI: 4.3-170.7]. A similar finding was obtained upon exclusion of patients with initial hyponatraemia with Fishers test [P=0.034; odds ratio (OR)=15.5, CI:1.7-140.6]. Binary logistic regression revealed a significantly increased in-hospital mortality in the hypochloraemic/normonatriaemic (OR=4.08, CI 1.08-15.43, P=0.039), but not in the hypochloraemic/hyponatraemic, normochloraemic/hyponatraemic or normonatriaemic/normochloraemic patients. Ejection fraction (EF) at admission was significantly higher in hypochloraemic/normonatriaemic, compared to normonatriaemic/normochloraemic patients, but similar to EF in both hypochloraemic/hyponatraemic and normochloraemic/hyponatraemic patients. The N-terminal precursor Brain Natriuretic Peptide (Nt-proBNP) levels at admission were significantly lower in hypochloraemic/normonatriaemic compared to hypochloraemic/hyponatraemic and normonatriaemic/normochloraemic patients, respectively. CONCLUSION: The data show that initial low serum chloride concentration is predictive of developing hyponatraemia and associated with increased in-hospital mortality in AHF patients.

Effects of simvastatin on malondialdehyde level and esterase activity in plasma and tissue of normolipidemic rats.

BACKGROUND: We investigated the possible non-lipid effects of simvastatin (SIMV) on paraoxonase 1 (PON1) and butyrylcholinesterase (BuChE) activity, as well as on malondialdehyde (MDA) levels in normolipidemic rats. METHODS: Two experimental groups of Wistar rats (10mg/kg/day of SIMV) and two control groups (saline) underwent a 21-day treatment period (TP). On the 22nd day one experimental and one control group of rats were sacrificed. Remaining groups of animals were sacrificied on the 32nd day of the study (10-day after-treatment period (AT)). Blood samples and slices of liver, heart, kidney, and brain tissue were obtained for the measurement of PON1 and BuChE activity and levels of MDA. Data were analyzed by means of t-test for independent samples. p values≤0.05 were considered as statistically significant. RESULTS: SIMV caused a significant decrease of serum and liver PON1 activity (18-24%, p≤0.05) and MDA concentrations in the plasma, heart, liver, kidney, and brain (9-40%, p≤0.05), while plasma and liver BuChE activity increased by 29% (p≤0.05) and 18%, respectively. All effects of SIMV were largely diminished following AT. The exception was MDA, which remained significantly decreased in plasma and all tissues analyzed. CONCLUSION: SIMV significantly decreased PON1 activity and MDA levels and increased BuChE activity. We suggest that the decrease of MDA levels is a beneficial therapeutic effect of SIMV, for example in cardiovascular disorders, while the increase of BuChE activity, especially in brain, may be a potential adverse effect in patients with Alzheimer disease.

5, 10-Methylenetetrahydrofolate reductase (MTHFR) 677 C --> T genetic polymorphism in 228 Croatian volunteers.

5, 10-Methylenetetrahydrofolate Reductase (MTHFR) is one of the key enzymes in the metabolism of homocysteine, where it catalyses its remethylation. The autosomal recessive bp 677 C --> T mutation in the MTHFR gene leads to the substitution of valine for alanine. Individuals who are homozygous for this C677T mutation exhibit a decreased specific activity and increased thermolability of this enzyme. This leads to increased plasma levels of homocysteine, which is a known risk factor for atherosclerosis and various manifestations of the atherosclerotic disease. The aim of this study was to find out the distribution and frequency of this mutation in the general Croatian population. A group of 228 volunteers (175 males and 53 females) has been analyzed for the MTHFR polymorphism, which revealed the following distribution: 105 (46.05%) individuals were without mutation (C/C), 102 (44.74%) were heterozygous (C/T) and 21 (9.21%) homozygous (T/T). These findings are within the results of studies on other European populations.

Folic acid and vitamin B(12) supplementation lowers plasma homocysteine but has no effect on serum bone turnover markers in elderly women: a randomized, double-blind, placebo-controlled trial.

An elevated homocysteine level is a newly recognized risk factor for osteoporosis. Older individuals may have elevated homocysteine levels due to inadequate folate intake and/or lower absorption of vitamin B(12). The aim of this study was to determine whether there is an impact of folic acid and vitamin B(12) supplementation on homocysteine levels and, subsequently, on bone turnover markers in older women with mildly to moderately elevated homocysteine levels. It is hypothesized that supplementation with folic acid and vitamin B(12) will improve homocysteine levels and, in turn, positively modify bone turnover markers in this population. This randomized, double-blind, placebo-controlled trial included 31 women (65 to 93 years) with homocysteine levels greater than 10 µmol/L. Participants were randomly assigned to receive either a daily folic acid (800 µg) and vitamin B(12) (1000 µg) (n = 17) or a matching placebo (n = 14) for 4 months. The results showed significantly lower homocysteine concentrations in the vitamin group compared to the placebo group (10.6 vs 18.5 µmol/L, P = .007). No significant difference in serum alkaline phosphatase or C-terminal cross-linking telopeptide of type I collagen was found between the vitamin and placebo groups before or after supplementation. The use of folic acid and vitamin B(12) as a dietary supplement to improve homocysteine levels could be beneficial for older women, but additional research must be conducted in a larger population and for a longer period to determine if there is an impact of supplementation on bone turnover markers or other indicators of bone health.