Diagnostic criteria for diabetes revisited: making use of combined criteria

Background  

Existing cut-offs for fasting plasma glucose (FPG) and post-load glucose (2hPG) criteria are not equivalent in the diagnosis of diabetes and glucose intolerance. Adjusting cut-offs of single measurements have not helped so we undertook this project to see if they could be complementary.     

Chemical modification of chlorinated microbial polyesters

Chlorination of microbial polyesters poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxyoctanoate) (PHO) was carried out by passing chlorine gas through their solutions. The chlorine contents in chlorinated PHB (PHB-Cl) and chlorinated PHO (PHO-Cl) were between 5.45 and 23.81 wt % and 28.09 and 39.09 wt %, respectively. Molecular weights of the chlorinated samples were in the range of between one-half to one-fourth of the original values because of hydrolysis during the chlorination process. Thermal properties of the PHO-Cl were dramatically changed with an increase in its glass transition (T(g) = 2 degrees C) and the melting transition (T(m)). The T(g) of PHB-Cl varied from -20 to 10 degrees C, and its T(m) decreased to 148 degrees C. The chlorinated poly(3-hydroxyalkanoate)s (PHA-Cl) were converted to their corresponding quaternary ammonium salts (PHA-N(+)R(3)), sodium sulfate salts (PHA-S), and phenyl derivatives (PHA-Ph). Cross-linked polymers were also formed by a Friedel-Crafts reaction between benzene and PHA-Cl. The modified PHO derivatives were characterized by (1)H NMR and (13)C NMR spectrometry, Fourier transform infrared spectroscopy, gel permeation chromatography, and differential scanning calorimetry techniques.     

Inter- and intra-individual variation in resting oxygen consumption in post-larvae of the giant freshwater prawn,Macrobrachium rosenbergii (De Man)

In a study of the factors that influence metabolic rate in the giant freshwater prawn Macrobrachium rosenbergii, resting oxygen consumption (ROC) was measured in 90 post-larvae ranging in size from 0.1 to 2.8 g. As in many other animal species, ROC was strongly negatively related to body weight. A stressful event (anaesthesia with or without tagging) caused a sharp increase in the ROC that disappeared over a time scale of hours. As has been demonstrated for other species of crustaceans, ROC was highest in prawns in the pre-moult stage. Individual differences in ROC among prawns handled in the same way and in the same moult stage persisted over a period of hours, but not over days. It remains unclear, therefore, whether early differences in resting metabolic rate can explain the conspicuous differences in growth rate that are found in this species during the first few weeks of life and that profoundly influence subsequent life history events.     

Polyethyleneimine-based immunopolyplex for targeted gene transfer in human lymphoma cell lines

Background

Specific and efficient delivery of genes into targeted cells is a priority objective in non‐viral gene therapy. Polyethyleneimine‐based polyplexes have been reported to be good non‐viral transfection reagents. However, polyplex‐mediated DNA delivery occurs through a non‐specific mechanism. This article reports the construction of an immunopolyplex, a targeted non‐viral vector based on a polyplex backbone, and its application in gene transfer over human lymphoma cell lines.

Methods

Targeting elements (biotin‐labeled antibodies), which should recognize a specific element of the target cell membrane and promote nucleic acid entry into the cell, were attached to the polyplex backbone through a bridge protein (streptavidin). Immunopolyplex transfection activity was studied in several hematological cell lines [Jurkat (CD3+/CD19?), Granta 519 (CD3?/ CD19+), and J.RT3‐T3.5 (CD3?/CD19?)] using the EGFP gene as a reporter gene and anti‐CD3 and anti‐CD19 antibodies as targeting elements. Transfection activity was evaluated via green fluorescence per cell and the percentage of positive cells determined by flow cytometry.

Results

A significant selectivity of gene delivery was observed, since the anti‐CD3 immunopolyplex worked only in Jurkat cells while the anti‐CD19 immunopolyplex worked only in the Granta cell line. Moreover, transfection of a CD3+/CD3? cell mixture with anti‐CD3 immunopolyplexes showed up to 16‐fold more transfection in CD3+ than in CD3? cells. Several non‐specific transfection reagents showed poor or no transfection activity.

Conclusion

It is concluded that immunopolyplex is a good non‐viral vector for specific and selective nucleic acid delivery. Immunopolyplex design allows easy replacement of the targeting element (antibody) – the streptavidin–polyplex backbone remaining intact – thereby conferring high versatility. Copyright © 2002 John Wiley & Sons, Ltd.

     

New action pattern of a maltose-forming alpha-amylase from Streptomyces sp. and its possible application in bakery

An a-Amylase (EC 3.2.1.1) was purified that catalyses the production of a high level of maltose from starch without the attendant production of glucose. The enzyme was produced extracellularly by thermophilic Streptomyces sp. that was isolated from Thailand's soil. Purification was achieved by alcohol precipitation, DEAE-Cellulose, and Gel filtration chromatographies. The purified enzyme exhibited maximum activity at pH 6-7 and 60 degrees C. It had a relative molecular mass of 45 kDa, as determined by SDS-PAGE. The hydrolysis products from starch had alpha-anomeric forms, as determined by 1H-NMR. This maltose-forming alpha-Amylase completely hydrolyzed the soluble starch to produce a high level of maltose, representing up to 90%. It hydrolyzed maltotetrose and maltotriose to primarily produce maltose (82% and 62% respectively) without the attendant production of glucose. The high maltose level as a final end-product from starch and maltooligosaccharides, and the unique action pattern of this enzyme, indicate an unusual maltose-forming system. After the addition of the enzyme in the bread-baking process, the bread's volume increased and kept its softness longer than when the bread had no enzyme.     

Periodic change in DO concentration for efficient poly-β-hydroxy-butyrate production using temperature-inducible recombinant<Emphasis Type="Italic">Escherichia coli</Emphasis> with proteome analysis

RecombinantEscherichia coli strain harboring the λp _R-p _L promotor and heterologus poly-β-hydroxybutyrate (PHB) biosynthesis genes was used to investigate the effect of culture conditions on the efficient PHB production. The expression ofphb genes was induced by a temperature upshift from 33°C to 38°C. The protein expression levels were measured by using two-dimensional electrophoresis, and the enzyme activities were also measured to understand the effect of culture temperature, carbon sources, and the dissolved oxygen (DO) concentration on the metabolic regulations. AcetylCoA is an important branch point for PHB production. The decrease in DO concentration lowers the citrate synthase activity, thus limit the flux toward the TCA cycle, and increase the flux for PHB production. Since NADPH is required for PHB production, the PHB production does not continue leading the overproduction of acetate and lactate. Based on these observations, a new operation was considered where DO concentration was changed periodically, and it was verified its usefulness for the efficient PHB production by experiments.