Application of green chemistry in decreasing adverse effect of (R,S)-ibuprofen

Lipases from Candida rugosa (OF and MY) were tested for their application in the enzymatic kinetic resolution of (R,S)-ibuprofen by enantioselective esterification. In this study, screening of enzymes was performed, and lipase MY was selected as an optimal catalyst, which allows to obtain products with high enantiopurity. Additionally, the influence of reaction time on the enantiomeric ratio and conversion was tested. High values of enantiomeric ratio (E in the range of 40.1–71.3) of the esterification of (R,S)- -ibuprofen were obtained using lipase MY, which has a great significance in the field of pharmaceutical synthesis of drugs. The chiral compounds (substrates and products) were analysed with the use of chiral stationary phases. As a result of the optimization, the reaction performed with the application of lipase MY allowed to achieve less toxic for human health (S)-enantiomer of ibuprofen with the high enantiomeric excess of product eep = 95%. Conversion of the reaction was c = 30.6% and enantioselectivity E = 58.9 after 126 h of incubation

Metabolic chiral inversion of 2-arylpropionic acid derivatives (profens)

2-arylpropionic acid derivatives (profens) are one of the most popular anti-inflammatory, analgesic, and antipyretic drugs. They belong to a group of nonsteroidal anti-inflammatory drugs (NSAID) and exhibit metabolic chiral inversion. Enantiomers of these chiral drugs are often characterised by different pharmacological activity. It is estimated that the values of metabolic chiral inversion of (R)-ibuprofen in humans are between 35 and 70%, depending on the condition of the liver and the intake of other medicines, while (R)-flurbiprofen undergoes chiral metabolic inversion to its opposed (S) form only in small range. The described phenomenon in the case of (R)-ketoprofen is limited to a maximum of around 10%. The metabolic chiral inversion is associated with potentially important pharmacotherapeutic and toxicological consequences, and so an attempt was made to analyse this phenomenon for the most commonly used drugs from the profens group

Biomarkers of bone cell activity in children and adolescents with newly diagnosed, untreated acute lymphoblastic leukemia

Introduction. Controversial data on disturbances in mineral homeostasis and bone mass were reported in children with diagnosed untreated acute lymphoblastic leukemia (ALL). Early detection of bone metabolism abnormalities is important for monitoring the effect of therapy on the skeleton. The purpose of this study was to evaluate bone metabolism in children and adolescents with newly diagnosed acute lymphoblastic leukemia by assessing biomarkers of bone cell activity. Materials and methods. Propeptide of type 1 procollagen (P1NP) and osteocalcin (OC) as bone formation markers and C-terminal telopeptide of type 1 collagen (CTX) and tartrate resistant acid phosphatase 5b (TRAP 5b) as resorption markers were determined in 22 Caucasian children and adolescents (12 boys 4–21 years, 10 girls 4–16 years) with newly diagnosed, untreated ALL and in 22 age- and gender-matched controls. Results. Bone formation, in particular, and bone resorption were significantly reduced in ALL children and adolescents compared with controls (Me P1NP 51.9 vs. 433.4 μg/L and OC 16.1 vs. 80.5 μg/L; p < 0.0001; Me CTX 0.454 vs. 1.225 μg/L and TRAP 5b 2.8 vs. 5.6 U/L; p < 0.001). P1NP positively correlated with OC (r = 0.56; p = 0.01) and CTX correlated with TRAP 5b (r = 0.54; p = 0.02) in children and adolescents with ALL. Median P1NP and OC concentrations in ALL children (4–9 years) were dramatically reduced compared with the healthy ones (10-fold and 9-fold respectively), whereas in adolescents with ALL (10–21 years) both bone formation markers were reduced in a lesser degree in comparison with the healthy adolescents. Conclusions. Acute lymphoblastic leukemia influences bone metabolism which is strongly related to the age of onset. More significant disturbances in bone turnover, particularly in bone formation (suppression of collagen synthesis), are observed in children with untreated ALL in comparison with adolescents with ALL.

Evaluation of Designed Immobilized Catalytic Systems: Activity Enhancement of Lipase B from Candida antarctica

Publisher's version (útgefin grein)Immobilized enzymatic catalysts are widely used in the chemical and pharmaceutical industries. As Candida antarctica lipase B (CALB) is one of the more commonly used biocatalysts, we attempted to design an optimal lipase-catalytic system. In order to do that, we investigated the enantioselectivity and lipolytic activity of CALB immobilized on 12 different supports. Immobilization of lipase on IB-D152 allowed us to achieve hyperactivation (178%) in lipolytic activity tests. Moreover, the conversion in enantioselective esterification increased 43-fold, when proceeding with lipase-immobilized on IB-S861. The immobilized form exhibited a constant high catalytic activity in the temperature range of 25 to 55°C. Additionally, the lipase immobilized on IBD152 exhibited a higher lipolytic activity in the pH range of 6 to 9 compared with the native form. Interestingly, our investigations showed that IB-S500 and IB-S60S offered a possibility of application in catalysis in both organic and aqueous solvents. A significant link between the reaction media, the substrates, the supports and the lipase was confirmed. In our enzymatic investigations, highperformance liquid chromatography (HPLC) and the titrimetric method, as well as the Bradford method were employed.This work was supported by the National Science Centre Poland grant DEC-2013/09/N/NZ7/03557.Peer Reviewe

Alpha-methylacyl-CoA racemase and hepsin as urinary prostate cancer markers

Background: Because of the numerous limitations of prostate-specific antigen (PSA), α-methylacyl-CoA racemase (AMACR) and hepsin have recently been suggested as potential biomarkers in prostate cancer (PC). This report presents a comparison study of the presence of AMACR and hepsin in urine collected before and after digital rectal examination (DRE) as a previously suggested diagnostic marker for PC. Methods: Seventy-six urine samples (38 before and 38 after prostate massage) from patients with benign prostatę hyperplasia (BPH) and 66 urine samples (33 before and 33 after prostate massage) from patients with PC were analyzed. PC was confirmed by prostate biopsy. Urinary levels of AMACR and hepsin were determined by ELISA and related to the tumor stage, Gleason score and PSA level. Results: AMACR and hepsin levels in urine collected after prostate massage were higher only in the PC group. There were no correlations between AMACR levels, hepsin levels, tumor stage and Gleason score. AMACR and hepsin did not differentiate between BPH and PC with better true positive and false negative rates than serum PSA. Conclusions: AMACR and hepsin were unable to diagnose PC with better true positive and false negative rates than PSA. An additional procedure combined with other markers should be applied for the reliable diagnosis of PC

Comparison of two different methods for routine 25(OH)D measurement in paediatric serum samples

Over the last decade interest in automated assays for 25-hydroxy-vitamin D measurement have greatly increased. The presence of different metabolites of vitamin D in the blood influences measurement of its concentration. In paediatric subjects the basic interference is due to the presence of 3-epi-25(OH)D2/D3, which  despite their biological inactivity, influences the total concentration of 25(OH)D. Aim: We assessed the analytical performance and usefulness of two different assays for measurement of total 25(OH)D in children. Materials and Methods: The study was performed in blood samples taken from 100 school-children aged 9–11 years. In all serum samples 25(OH)D total concentration was measured with the use of chemilumi-nescent assay, which is known to show no cross-reactivity with 3-epi-25(OH)D, and with the use of a newly developed enzyme-immunosorbent method. Results: The mean 25(OH)D concentration in children measured with enzyme-immunosorbent assay (EIA) was significantly higher, at 28.06 ng/mL, than with the chemiluminescent assay (CLIA), at 21.13 ng/mL;   < 0.0001. In children with optimal weight the average 25(OH)D was 32.93 ng/ml (EIA) and 21.5 ng/mL (CLIA) (p < 0.0001), respectively, whereas in a subgroup with overweight/obesity the mean concentra-tion of 25(OH)D was similar, at 23.2 ng/ml (EIA) and 20.76 ng/ml (CLIA) (p = 0.15). The nonparametric Spearman’s rank correlation of two methods equalled 0.47; 95%CI (0.11 to 0.60) with a significance level  p < 0.0001. The calculated concordance correlation coefficient between two methods in the whole group was 0.26; 95%CI (0.17 to 0.35). In a subgroup of children with optimal body mass (N = 50) the concor-dance correlation coefficient was 0.18; 95%CI (0.06 to 0.29), whereas in children with overweight/obesity (N = 50) it was 0.44; 95%CI (0.29 to 057). Mean bias for the enzyme-immunosorbent method equalled 18.7%; +/- 1.96 SD (101.3% to -64%). Conclusions: With reference to 25(OH)D measurement in children, Spearman’s correlation coefficient indicated “moderate correlation” between the two compared methods, whereas the strength of agree-ment (concordance) between both methods was characterised as “poor”. The proper selection of assay for accurate assessment of vitamin D status in paediatric samples is necessary to avoid misdiagnosis

Serum 25(OH)D status and lipid profile in children with newly diagnosed asthma

Background. The problem of the influence of hyperlipidemia on asthma was addressed several years ago. Systematic review and meta-analysis performed in the pediatric population on the association between vitamin D status and lipid profile components revealed discordant results and indicated that higher serum 25(OH)D is related to a more favorable lipid profile. Objective. We aimed to elucidate whether there was an association between vitamin D status and lipid profile components and apolipoprotein B in a sample of children with newly diagnosed atopic asthma. Methods. The study included 150 children aged 2–12 years. Atopic asthma was diagnosed in 110 children; 40 children constituted a reference group. Fasting blood was collected to measure 25(OH)D total, lipid profile and apolipoprotein B concentrations. Results. Children with asthma had significantly lower 25(OH)D (p < 0.0001) but similar lipid and apolipoprotein B concentrations. The proportions of hypercholesterolemia, hypertriglyceridemia and increased apoB concentrations were similar in both groups. HDL-C concentrations in asthmatic 25(OH)D-deficient children were higher compared with the children with sufficient levels (p = 0.05). ApoB concentration was lower in 25(OH)D-deficient compared with vitamin D sufficient asthmatics (p = 0.0008). Correlations between 25(OH)D concentration and lipids and apoB in asthmatics revealed gender differences. An inverse relationships between vitamin D and total cholesterol and HDL-C (R= –0.39, p < 0.05; R= –0.475, p < 0.001) were found in girls. In boys vitamin D correlated with LDL-C and apoB (R = 0.376, R = 0.498; p < 0.001). Conclusion. In children with asthma lower 25(OH)D had more favorable gender-dependent effect on the lipid profile. The association of serum 25(OH)D and lipid levels in children with asthma remains for further studies.

Engrailed-2 protein as a potential urinary prostate cancer biomarker: a comparison study before and after digital rectal examination

This study was designed to compare and evaluate the presence of engrailed-2 (EN2) protein in urine collected before and after prostate massage as a diagnostic marker for prostate cancer (PCa). We analysed and compared 76 urine samples (38 before and 38 after prostate massage) from the benign group (BPH) and 66 urine samples (33 before and 33 after prostate massage) from patients with PCa confirmed by prostate biopsy. EN2 levels from the PCa and men with BPH (age range 50–82) were related to the tumour stage, Gleason score and prostate-specific antigen. EN2 levels were determined by enzyme-linked immunosorbent assay in urine. The median EN2 levels in urine after prostate massage were significantly different from those determined in urine before prostate massage (1.25 ng/ml in the PCa group and 0.34 ng/ml in the BPH). The mean EN2 levels in PCa patients were 3.76-fold higher than those in non-PCa patients after prostate massage. The distinct influence of prostate massage on EN2 levels was found to be related to the Gleason score and tumour stage. EN2 may be considered a marker of PCa with certain limitations, such as those related to tumour staging. The specificity and sensitivity of the protocol are highly dependent on prostate massag

25-hydroxyvitamin D insufficiency in children with newly diagnosed asthma

Background. 25-hydroxyvitamin D [25(OH)D] deficiency seems to be related to the development of asthma. Any evaluation of the relationship between asthma and 25(OH)D deficiency must consider the association between increased airway responsiveness, eosinophil counts and serum immunoglobulin E (IgE), and 25(OH)D as a potential player in airway remodelling. Objective. We assessed the association of 25(OH)D with markers of atopy and eosinophilic inflammation in children with newly diagnosed asthma. Methods. The study included 165 children aged 2–12 years. The diagnosis of asthma was performed by an experienced paediatric pulmonologist. Allergic asthma was diagnosed in 106 children, and non-allergic asthma in ten; in 49 children, asthma was excluded. Fasting blood was collected for cell counts, and serum was obtained to measure lipids, C-reactive protein (hsCRP), 25(OH)D and IgE. Results. Children with asthma had significantly lower 25(OH)D (p < 0.001). Both groups had similar lipid values. Elevated total IgE concentration and eosinophil counts were found in asthmatics; neutrophils were similar in asthmatic and reference groups. There was a strong tendency to higher eosinophil counts in 25(OH)D-deficient children (< 20 ng/mL) with atopic asthma (p < 0.08). Conclusion. In children with asthma, 25(OH)D insufficiency/deficiency is associated with higher eosinophil counts and IgE. 25(OH)D monitoring is important in the prevention and management of children with asthm

The occurrence of cardiovascular risk factors and functioning in chronic illness in the Polish population of EUROASPIRE V

Background: The aim of this study was to assess the impact of cardiovascular risk on the functioning of patients without a history of atherosclerotic cardiovascular disease. Methods: Two hundred patients diagnosed with arterial hypertension, hypercholesterolemia, or diabetes were enrolled in the study. The median age was 52.0 years (interquartile range [IQR] 43.0–60.0). The following risk factors were assessed: blood pressure, body mass index, waist circumference, physical activity, smoking, LDL-cholesterol, triglycerides, and fasting plasma glucose concentration. Total cardiovascular risk was determined as the number of uncontrolled risk factors, and with the Systemic Coronary Risk Evaluation Score (SCORE). The Functioning in the Chronic Illness Scale (FCIS) was applied to assess the physical and mental functioning of patients. Results: The median number of measures of cardiovascular risk factors was 4.0 (IQR 3.0–5.0). The median of SCORE for the whole study population was 2.0 (IQR 1.0–3.0). Patients with lower total cardiovascular risk as defined by SCORE and number of uncontrolled risk factors had better functioning as reflected by higher FCIS (R = –0.315, p < 0.0001; R = –0.336, p < 0.0001, respectively). Multivariate logistic regression analysis identified abnormal blood pressure, abnormal waist circumference, tobacco smoking, and lack of regular physical activity to be negative predictors of functioning. Lack of regular physical activity was the only predictor of low FCIS total score (odds ratio 9.26, 95% confidence interval 1.19–71.77, p = 0.03). Conclusions: The functioning of patients worsens as the total cardiovascular risk increases. Each of the risk factors affects the functioning of subjects without coronary artery disease with different strength, with physical activity being the strongest determinant of patient functioning