Quantification of Mixtures of C2 and C3 Hydroxy Acids and Products of Glycerol Oxidation by High-Performance Liquid Chromatography and Quantitative 13C NMR

The aim of this study was to develop an improved general method for detecting and quantifying mixtures of hydroxy acids and other products of glycerol oxidation in aqueous media, to prevent the confusions that can occur due to similarities and interactions between these compounds depending on media conditions. Standard potential products of glycerol oxidation—glycerol, glyceraldehyde, dihydroxyacetone, glyceric acid, lactic acid, glycolic acid, glyoxylic acid, oxalic acid, tartronic acid, and mesoxalic acid—were analyzed by high-performance liquid chromatography (HPLC) and quantitative ¹³C nuclear magnetic resonance (NMR), in mixtures of known composition. The results obtained were concordant with the known compositions tested. HPLC was more accurate than quantitative ¹³C NMR for simple mixtures, but ¹³C NMR was required for complex mixtures containing dihydroxyacetone and glycerol, oxalic acid and mesoxalic acid, or glyoxylic acid and tartronic acid, pairs of compounds not well separated or detected by HPLC. As proof-of-concept, an unknown mixture generated by glycerol oxidation was analyzed by HPLC and quantitative ¹³C NMR. The results obtained were concordant and allowed accurate determination of the composition of the sample, which contained mesoxalic acid as the major product, with oxalic acid, tartronic acid, and glyceric acid as by-products.     

APOB CRISPR-Cas9 Engineering in Hypobetalipoproteinemia: A Promising Tool for Functional Studies of Novel Variants

Hypobetalipoproteinemia is characterized by LDL-cholesterol and apolipoprotein B (apoB) plasma levels below the fifth percentile for age and sex. Familial hypobetalipoproteinemia (FHBL) is mostly caused by premature termination codons in the APOB gene, a condition associated with fatty liver and steatohepatitis. Nevertheless, many families with a FHBL phenotype carry APOB missense variants of uncertain significance (VUS). We here aimed to develop a proof-of-principle experiment to assess the pathogenicity of VUS using the genome editing of human liver cells. We identified a novel heterozygous APOB-VUS (p.Leu351Arg), in a FHBL family. We generated APOB knock-out (KO) and APOB-p.Leu351Arg knock-in Huh7 cells using CRISPR-Cas9 technology and studied the APOB expression, synthesis and secretion by digital droplet PCR and ELISA quantification. The APOB expression was decreased by 70% in the heterozygous APOB-KO cells and almost abolished in the homozygous-KO cells, with a consistent decrease in apoB production and secretion. The APOB-p.Leu351Arg homozygous cells presented with a 40% decreased APOB expression and undetectable apoB levels in cellular extracts and supernatant. Thus, the p.Leu351Arg affected the apoB secretion, which led us to classify this new variant as likely pathogenic and to set up a hepatic follow-up in this family. Therefore, the functional assessment of APOB-missense variants, using gene-editing technologies, will lead to improvements in the molecular diagnosis of FHBL and the personalized follow-up of these patients.     

Silicon-doped hydroxyapatite prepared by a thermal technique for hard tissue engineering applications

Hydroxyapatite (HA, Ca₅(PO₄)₃OH) has been extensively used for bone implantation due to its similarity to the mineral component of bone, which makes it strongly osteoconductive. However, HA has low resorbability, and it is difficult to replace by a newly regenerated bone. Si doping can enhance the resorbability of HA by modifying its crystal structure. Here, we developed a simple thermal technique for preparing Si-doped HA from silica (SiO₂) and HA precursors, both of which are inexpensive and commercially available. This method included the physical binding of SiO₂ and HA particles, followed by pressing and sintering the mixture at an elevated temperature, which enhanced the atomic diffusion of Si into HA unit cells. We also evaluated the simulated body fluid (SBF) activity of the Si-doped HA prepared by this technique and showed that it significantly had higher resorbability and mineralizing potential compared to the pure HA. Our experimental design including, the individual precipitation and resorption assays enabled us to explain the mechanism behind the improved activity of Si-doped HA in SBF. This was attributed to the formation of new phases, such as β-tricalcium phosphate (β-TCP) and calcium silicate (Ca₂SiO₄) with higher solubility than HA on the SiO₂-contating HA during the sintering stage. This can provide some guidelines for designing new calcium phosphate-based materials for hard tissue engineering applications.     

Mid-infrared spectroscopy as a tool for rapid determination of internal quality parameters in tomato

The objective of this study was to investigate the possibility of predicting the quality parameters of tomato by mid-infrared spectroscopy. For 2 years, tomato samples, representing a large variability in the chemical composition, were scanned using the attenuated total reflectance accessory of a Fourier transform spectrometer in the wavenumber region between 4000 and 400 cm⁻¹_. Calibration models were developed using partial least squares (PLS) regression method and were tested with internal validation sample set in the first year. Different spectral preprocessing techniques were investigated and different spectral regions were selected to optimise the calibration models. In addition, the models obtained in 2007 were used to predict the soluble solids, dry matter and total acidity in tomato harvested in 2008.     

Amino‐functionalised mesoporous silica microspheres for immobilisation of Candida antarctica lipase B – application towards greener production of 2,5‐furandicarboxylic acid

In the present study, amino‐functionalised mesoporous silica microspheres were utilised as support for the covalent immobilisation of Candida antarctica lipase B (CaLB) for the subsequent production of 2,5‐furandicarboxylic acid (FDCA) from 2,5‐diformylfuran (DFF). Under the optimised operating conditions of pH 6.5, particle/enzyme ratio of 1.25:1.0 and glutaraldehyde concentration of 4 mM, a maximum CaLB immobilisation yield of 82.4% on silica microspheres was obtained in 12.25 h. The immobilised CaLB was used for the synthesis of alkyl esters, which were utilised along with hydrogen peroxide for FDCA synthesis. The biocatalytic conversion of 30 mM DFF dictated a 77–79% FDCA in 48 h at 30°C; where the turnover number and turnover frequency of immobilised CaLB were 6220.73 mol mol⁻¹ and 129.59 h⁻¹, respectively, for ethyl acetate, against 6297.65 mol mol⁻¹ and 131.2 h⁻¹, respectively, for ethyl butyrate. Upon examining the operational stability, the immobilised CaLB exhibited high stability till five cycles of FDCA production.Inspec keywords: mesoporous materials, organic compounds, biotechnology, silicon compounds, renewable materials, catalysis, catalysts, enzymes, thermal stability, biofuelOther keywords: FDCA production, amino‐functionalised mesoporous silica microspheres, greener production, 2,5‐furandicarboxylic acid, covalent immobilisation, 2,5‐diformylfuran, CaLB immobilisation, Candida antarctica lipase B immobilisation, ethyl butyrate, time 48.0 hour, temperature 30.0 degC, time 12.25 hour     

UNICORE 6 — Recent and Future Advancements

UNICORE is a European Grid Technology with more than 10 years of history. Originating from the Supercomputing domain, the latest version UNICORE 6 has turned into a general-purpose Grid technology that follows established standards and offers a rich set of features to its users. The paper starts with an architectural insight into UNICORE 6, highlighting the workflow features, standards and the different clients. Next, the current state of advancement is presented by describing recent developments. The paper closes with an outlook on future planned developments.     

Complexation Hydrogels for the Oral Delivery of Growth Hormone and Salmon Calcitonin

The hydrogel system of poly(methacrylic acid-co-N-vinyl pyrrolidone) was evaluated for use as an oral delivery system for growth hormone and salmon calcitonin. These proteins were selected because of their therapeutic importance and the insight provided by evaluating the delivery of a therapeutic agent with a high molecular weight (growth hormone) and a drug with a high isoelectric point (salmon calcitonin). Growth hormone loading and release studies were performed for both P(MAA-co-NVP) and P(MAA-g-PEG). Loading efficiencies for the respective systems were 50.9 ± 1.8% and 57.8 ± 4.1%; weight incorporation of the protein was determined to be 3.5 ± 0.1% and 4.0 ± 0.3%. At pH 7.4, growth hormone release of 90% occurred within 45 min for P(MAA-co-NVP) microparticles; 90% release was not achieved with P(MAA-g-PEG) microparticles until 180 min. At pH 1.2, no release occurred from P(MAA-co-NVP) microparticles but 10% release occurred from P(MAA-g-PEG) microparticles. Salmon calcitonin loading and release were shown to be affected by the negative charges of deprotonated MAA; for systems with monomer molar feed ratios of 4:1, 1:1 and 1:4 MAA:NVP, loading efficiencies were determined to be 70.6 ± 3.0%, 25.3 ± 1.2%, and 1.6 ± 1.3%. Salmon calcitonin release was minimal from the copolymer with 4:1 MAA:NVP monomer feed at pH 7.4. The release improved when the pH was raised above physiological levels. These studies confirmed that P(MAA-co-NVP) was an effective oral delivery system for high molecular weight drugs, but improvements are needed before the system could be utilized for high isoelectric point therapeutic delivery.     

Second-order facial information processing in schizophrenia.

This study investigated the processing of second-order relational face information in schizophrenia. Twenty-eight schizophrenic patients and 28 controls were asked to say whether the space between the eyes was the same in 2 side-by-side faces. The 2 faces were derived from the same original face, but the spacing between the eyes was either the same or differed by various distances. The results showed that schizophrenic patients needed a space that was twice as great as controls to see a difference. The authors conclude that schizophrenic patients have a deficit in processing second-order relational face information. (PsycINFO Database Record (c) 2010 APA, all rights reserved)