Modeling and evaluation of the sucrose-degrading activity of recombinantly produced oligo-1,6-glucosidase from A. gonensis.

Fructose is the most preferred sugar to provide benefits for sweetening and health. As many industrial enzymes are used to produce High Fructose Syrup (HFS), it is vital to explore alternative enzymes for fructose production. Oligo-α-1,6-glucosidase (O-1-6-glucosidase) hydrolyzes non-reducing ends of isomaltooligosaccharides, panose, palatinose, and an a-limit dextrin by breaking α-1,6-glucoside bonds, although it generally has no activity on α-1,4-glucoside bonds of maltooligosaccharides. In this study, sucrose-hydrolyzing activity of O-1-6-glucosidase of thermophilic A. gonensis was evaluated. For this purpose, O-1-6-glucosidase gene region of A. gonensis was cloned in the pET28(a)+ expression vector, the expression product was purified, modeled, and biochemically characterized. The optimal activity of the enzyme was to be at pH 7.0 and 60 °C. The enzyme activity was halved at the end of the 276th h at 60 °C. The enzyme maintained its activity even after 300 h at pH 6.0-10.0. The values of Km, Vmax, kcat, and kcat/Km were determined as 44.69 ± 1.27 mM, 6.28 ± 0.05 µmoL/min/mg protein, 6.70 1/s and 0.15 1/mMs-1, respectively. While Zn2+, Cu2+, Pb2+, Ag2+, Fe3+, Hg2+, and Al2+ metal ions inhibited O-1-6-glucosidase, Mn2+, Fe2+, and Mg2+ ions activated the enzyme. Consequently, A. gonensis O-1-6-glucosidase (rAgoSuc2) has interesting properties, especially for HFS production.

Risk estimation for biliary tract cancer: Development and validation of a prognostic score.

BACKGROUND & AIMS: Biliary tract cancer is a rare tumour entity characterized by a poor prognosis. We aimed to identify prognostic factors and create a prognostic score to estimate survival. METHODS: Clinical data of the training set, consisting of 569 patients treated from 2000 to 2010 at Hannover Medical School, were analysed. A prognostic model defining three prognostic risk groups was derived from Cox regression analyses. The score was applied and validated in an independent cohort of 557 patients from four different German centres. RESULTS: Median overall survival (OS) was 14.5 months. If complete resection was performed, the patients had a significantly improved OS (23.9 months; n=242) as compared to patients with non-resectable tumours (9.1 months; n=329, P<.0001). Based on univariable and multivariable analyses of clinical data, a prognostic model was created using variables available before treatment. Those were age, metastasis, C-reactive protein (CRP), international normalized ratio (INR) and bilirubin. The prognostic score distinguished three groups with a median OS of 21.8, 8.6 and 2.6 months respectively. The validation cohort had a median OS of 20.2, 14.0 and 6.5 months respectively. The prognostic impact of the score was independent of the tumour site and of treatment procedures. CONCLUSIONS: Here, we identified prognostic factors and propose a prognostic score to estimate survival, which can be applied to all patients independent of tumour site and before initial treatment. Further validation in prospective trials is required.

Childhood obesity-related cardiovascular risk factors and carotid intima-media thickness.

The purpose of this study was to investigate the relationship between childhood obesity and carotid intima-media thickness (IMT). This is a cross-sectional study in obese children and non-obese control subjects. This study included 75 obese children and 40 non-obese control children. Systolic and diastolic blood pressure (SBP, DBP) values and waist and hip circumferences were measured. Fasting blood glucose and insulin concentrations, total cholesterol, triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were assayed. The carotid IMT was measured by high resolution B-mode ultrasonography. Waist/hip ratios, SBP and DBP were significantly increased in the obese group compared to the non-obese children (all p < 0.001). The total cholesterol, LDL-C, HDL-C, and TG in the obese children were significantly different from values in the control subjects (all p < 0.001). Compared to the controls, the obese children demonstrated significant differences in a number of clinical risk factors including body weight, body mass index (BMI), BMI-standard deviation score (SDS), SBP/DBP, waist circumference, hip circumference, and waist/hip ratio (all p < 0.001). Compared to the controls, the obese children showed increased mean carotid IMT values [0.52 mm (95% confidence interval [CI], 0.40-0.64 mm) vs. 0.35 mm (95% CI, 0.24-0.38 mm), p < 0.001]. Univariate correlation analysis revealed that the carotid IMT was closely related to the BMI-SDS, SBP/DBP, waist and hip circumferences, serum TG, cholesterol, LDL-C, HDL-C, fasting serum insulin level, and insulin resistance indices including the homeostasis model assessment of insulin resistance (HOMA-IR), fasting glucose-to-insulin ratio (FGIR), and quantitative insulin-sensitivity check index (QUICKI). Multiple regression analysis showed that the BMI-SDS, TG and QUICKI were independent predictive risk factors for increased carotid IMT. Measurements of BMI-SDS, blood pressure, waist and hip circumferences, serum TG levels, the QUICKI insulin resistance index, and carotid IMT by ultrasonography are suitable in pediatric patients in a clinical setting and may be used for screening of obese children.

Solvothermal synthesis of WO3 /TiO2 /carbon fiber composite photocatalysts for enhanced performance under sunlight illumination.

Carbon fiber (CF)-based WO3 /TiO2 composite catalysts (WO3 /TiO2 /CF) were successfully synthesized by solvothermal method. The catalysts were characterized by XPS, SEM, BET, XRD, FTIR, Raman and UV-Vis. The analyses confirmed the WO3 /TiO2 nanoparticles with high crystallinity deposited on the carbon structure. The photocatalytic degradation of Orange II azo dye under UV and sunlight illumination with the synthesized catalyst was explored. The composite catalyst displayed high performance (85%) for Orange II degradation while that of for WO3 /TiO2 was found as 76%. The effects of CF amount, solution pH, initial dye concentration and catalyst dose on photocatalytic performance were studied. It was found that the degradation efficiency increased from 68% to 90% with the increasing CF amount from 3 wt% to 5 wt%, while the further increase in CF amount (7-10 wt%) decreased the photodegradation due to the blocking the active sites of WO3 /TiO2 . The enhanced photocatalytic efficiency was mainly attributed to the electrical properties of the CF and reduced bandgap.

Hodgkin's lymphoma in a patient with Jo-1 syndrome.

Jo-1 syndrome is an autoimmune disease with autoantibodies against the histidyl tRNA synthetase. Characteristic clinical findings include inflammatory myopathy and interstitial lung disease. We present the first case of a patient with Jo-1 syndrome (positive Jo-1 autoantibodies, myositis, interstitial alveolitis) who developed Hodgkin's lymphoma of nodular-sclerosing type. Thus, patients with Jo-1 syndrome and immunosuppressive therapy similar to other patients with autoimmune disease are at risk to develop lymphomas and should therefore be monitored carefully.

Determination of total antioxidant capacity of humic acids using CUPRAC, Folin-Ciocalteu, noble metal nanoparticle- and solid-liquid extraction-based methods.

Total antioxidant capacity (TAC) of humic acid (HA) samples was determined using CUPRAC (CUPric Reducing Antioxidant Capacity), FC (Folin-Ciocalteu), QUENCHER-CUPRAC, QUENCHER-FC, Ag-NP (Silver nanoparticle)‒ and Au-NP (Gold nanoparticle)‒based methods. Conventional FC and modified FC (MFC) methods were applied to solid samples. Because of decreased solubility of Folin-Ciocalteu's phenol reagent in organic solvents, solvent effect on TAC measurement was investigated using QUENCHER-CUPRAC assay by using ethanol:distilled water and dimethyl sulfoxide:distilled water with varying ratios. To see the combined effect of solubilization (leaching) and TAC measurement of humic acids simultaneously, QUENCHER experiments were performed at 25°C and 50°C; QUENCHER-CUPRAC and QUENCHER-FC methods agreed well and had similar precision in F-statistics. Although the Gibbs free energy change (ΔG°) of the oxidation of HA dihydroxy phenols with the test reagents were negative, the ΔG° was positive only for the reaction of CUPRAC reagent with isolated monohydric phenols, showing CUPRAC selectivity toward polyphenolic antioxidants. This is the first work on the antioxidant capacity measurement of HA having a sparingly soluble matrix where enhanced solubilization of bound phenolics is achieved with coupled oxidation by TAC reagents.

Extracorporeal Treatment in Severe Hypertriglyceridemia-Induced Pancreatitis.

Plasmapheresis is a well-accepted treatment option in severe hypertriglyceridemia-induced pancreatitis (HTGP). The rationale behind this approach is the depletion of triglycerides and the reduction of inflammatory cytokines. The time span between onset of clinical symptoms and start of plasmapheresis might have an important impact on mortality. Hyperviscosity of patients' plasma represents another special challenge for the applied separation technology. The procedures can be performed either by centrifugal device (CFD) or membrane based (MBS) units. The present study reports the outcome of 10 patients suffering from HTG. The expected mortality of the collective was 25%. Plasmapheresis was started after an average 16.3 h (SD ± 6.7 h) after onset of symptoms. No mortality occurred. Apheresis was statistically equally effective with both devices. A median of 3 sessions reduced the TG level to normal and correlated with patients' improvement. During follow up, three patients developed a pancreatic pseudocyst requiring surgical intervention without further complication.