Which criteria should be used for starting pharmacologic therapy for management of gestational diabetes in pregnancy? Evidence from randomized controlled trials

Introduction: There is inconclusive evidence to support any specific criteria for starting pharmacologic therapy after diet in women with gestational diabetes mellitus (GDM). We aimed to analyze the most used criteria for starting pharmacologic treatment for patients with GDM. Material and methods: Electronic databases were searched from their inception to September 2017. We included all the randomized controlled trials (RCTs) of GDM managed initially by diet and exercise reporting criteria for starting pharmacologic therapy. RCTs in women with pregestational diabetes were excluded. Data regarding glucose values used for starting pharmacologic therapy were extracted and carefully reviewed. Results: We included 15 RCTs (4307 women) in the meta-analysis. For fasting glucose target, 8/14 (57%) used a value lower or equal to 90 mg/dL and the remainder used values 50% of the values higher than the target values and another one (7%) used >30%. Conclusion: The majority of RCTs (87%) used very tight criteria of either 1 or 2 values over the target values in the 1 or 2-week period for starting pharmacologic treatment for patients with GDM; more than 50% used 2 values. Key Message Pharmacologic therapy should be considered in women with gestational diabetes when, despite an adequate diet and exercise, 1 or 2 blood glucose values are over the target values of 90mg/dL fasting or 120mg/dL 2-hour postprandial over 1 or 2 weeks

Cellobiohydrolase B of Aspergillus niger over-expressed in Pichia pastoris stimulates hydrolysis of oil palm empty fruit bunches

Background. Aspergillus niger, along with many other lignocellulolytic fungi, has been widely used as a commercial workhorse for cellulase production. A fungal cellulase system generally includes three major classes of enzymes i.e.,β-glucosidases, endoglucanases and cellobiohydrolases. Cellobiohydrolases (CBH) are vital to the degradation of crystalline cellulose present in lignocellulosic biomass. However, A. niger naturally secretes low levels of CBH. Hence, recombinant production of A. niger CBH is desirable to increase CBH production yield and also to allow biochemical characterisation of the recombinant CBH from A. niger. Methods. In this study, the gene encoding a cellobiohydrolase B (cbhB) from A. niger ATCC 10574 was cloned and expressed in the methylotrophic yeast Pichia pastoris X-33. The recombinant CBHB was purified and characterised to study its biochemical and kinetic characteristics. To evaluate the potential of CBHB in assisting biomass conversion, CBHB was supplemented into a commercial cellulase preparation (Cellic ® CTec2) and was used to hydrolyse oil palm empty fruit bunch (OPEFB), one of the most abundant lignocellulosic waste from the palm oil industry. To attain maximum saccharification, enzyme loadings were optimised by response surface methodology and the optimum point was validated experimentally. Hydrolysed OPEFB samples were analysed using attenuated total reflectance FTIR spectroscopy (ATR-FTIR) to screen for any compositional changes upon enzymatic treatment. Results. Recombinant CBHB was over-expressed as a hyperglycosylated protein attached to N-glycans. CBHB was enzymatically active towards soluble substrates such as 4-methylumbelliferylβ-D-cellobioside (MUC), p-nitrophenyl-cellobioside (pNPC) and p-nitrophenyl-cellobiotrioside (pNPG3) but was not active towards crystalline substrates like Avicel ® and Sigmacell cellulose. Characterisation of purified CBHB using MUC as the model substrate revealed that optimum catalysis occurred at 50 °C and pH 4 but the enzyme was stable between pH 3 to 10 and 30 to 80 °C. Although CBHB on its own was unable to digest crystalline substrates, supplementation of CBHB (0.37%) with Cellic ® CTec2 (30%) increased saccharification of OPEFB by 27%. Compositional analyses of the treated OPEFB samples revealed that CBHB supplementation reduced peak intensities of both crystalline cellulose Iα and Iβ in the treated OPEFB samples. Discussion. Since CBHB alone was inactive against crystalline cellulose these data suggested that it might work synergistically with other components of Cellic R CTec2. CBHB supplements were desirable as they further increased hydrolysis of OPEFB when the performance of Cellic® CTec2 was theoretically capped at an enzyme loading of 34% in this study. Hence, A. niger CBHB was identified as a potential supplementary enzyme for the enzymatic hydrolysis of OPEFB

Bioinformatics and molecular modeling in glycobiology

The field of glycobiology is concerned with the study of the structure, properties, and biological functions of the family of biomolecules called carbohydrates. Bioinformatics for glycobiology is a particularly challenging field, because carbohydrates exhibit a high structural diversity and their chains are often branched. Significant improvements in experimental analytical methods over recent years have led to a tremendous increase in the amount of carbohydrate structure data generated. Consequently, the availability of databases and tools to store, retrieve and analyze these data in an efficient way is of fundamental importance to progress in glycobiology. In this review, the various graphical representations and sequence formats of carbohydrates are introduced, and an overview of newly developed databases, the latest developments in sequence alignment and data mining, and tools to support experimental glycan analysis are presented. Finally, the field of structural glycoinformatics and molecular modeling of carbohydrates, glycoproteins, and protein–carbohydrate interaction are reviewed

Optimization of callus and cell suspension cultures of Barringtonia racemosa (Lecythidaceae family) for lycopene production

Lycopene is present in a range of fresh fruits and vegetables, especially in the leaves of Barringtonia racemosa. The traditional lycopene extraction from the plant is being employed instead of an easy propagation technique like cell culture process from the leaf explants. We intend to assess how lycopene could be extracted via tissue culture under light (illuminance: 8,200 lux under white fluorescent lamps, photoperiod 16 h per day at 25ºC) and dark. Leaf explants of Barringtonia racemosa were cultured on modified Murashige and Skoog (MS), Woody Plant Medium (WPM) and B5 media, supplemented with different concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D). Optimal conditions for callus induction and maintenance under both dark and light were investigated, and growth and lycopene accumulation were evaluated. Among media with different concentrations of 2,4-D, fast growing, friable callus initiated within three weeks after culturing on WPM basal medium supplemented with 2.0 mg L-1 (weight per volume) of 2,4-D, whereas callus induction in explants cultured on all other media started only after five weeks. Calli were subcultured once every fortnight. Pale yellow and green calli developed under conditions of dark and light respectively were then selected for evaluation of their lycopene contents. An improved reversed phase of high performance liquid chromatography (HPLC) method was used for a selective chemical determination of the lycopene content. Light induced lycopene production; and likewise maximum lycopene level incubated in light was higher than those incubated in darkness. The best growth rates of callus and cell suspension were achieved in WPM and B5 media respectively. The production of lycopene was growth-dependent through analysis of growth and lycopene content of both callus and cell suspension cultures.O licopeno está presente numa série de frutas frescas e hortaliças principalmente na folhas de Barringtonia racemosa. A extração tradicional do licopeno tem sido empregada no lugar da fácil técnica de propagação como o processo de cultura de células de explantes de folhas. É nossa intenção demonstrar como o licopeno pode ser extraído através de cultura de tecido sob luz (iluminação com lâmpadas fluorescentes brancas de 8.200 lux, 16 h por dia a 25º C) e escuro. Explantes de folhas de Barringtonia racemosa foram cultivados em meio modificado de Murashige e Skoog (MS) para plantas lenhosas e meio B5, suplementado com diferentes concentrações de ácido 2,4-Diclorofenoxiacético (2,4-D). Condições ótimas para indução e manutenção de calos sob luz e escuro foram investigadas e avaliados o crescimento e acumulo de licopeno. Entre meios com diferentes concentrações de 2,4 -D, calos friáveis de crescimento rápido tiveram início em três semanas após serem cultivados em meio basal WPM suplementado com 2.0 mg L-1 (peso por volume) de 2,4-D enquanto indução de calos em explantes cultivados em todos os outros meios começaram somente após cinco semanas. Calos foram subrepicados a cada 15 dias. Calos amarelo-pálido e verdes desenvolvidos respectivamente sob condições escura e de luz foram então selecionados para avaliação do teor de licopeno. Um método aperfeiçoado de cromatografia líquida de alto desempenho foi usado para a determinação química seletiva do teor de licopeno. A produção de licopeno induzida sob luz e também o nível máximo de licopeno incubado em luz foi mais alto do que aqueles incubados no escuro. As melhores taxas de crescimento de calo e suspensões de células foram obtidas respectivamente em meio WPM e B5. A produção de licopeno dependeu do crescimento como demonstrado pela análise do crescimento e teor de licopeno de ambos calos e cultura de células em suspensão