Production of human recombinant proapolipoprotein A-I in Escherichia coli: purification and biochemical characterization

A human liver cDNA library was used to isolate a clone coding for apolipoprotein A-I (Apo A-I). The clone carries the sequence for the prepeptide (18 amino acids), the propeptide (6 amino acids), and the mature protein (243 amino acids). A coding cassette for the proapo A-I molecule was reconstructed by fusing synthetic sequences, chosen to optimize expression and specifying the amino-terminal methionine and amino acids -6 to +14, to a large fragment of the cDNA coding for amino acids 15-243. The module was expressed in pOTS-Nco, an Escherichia coli expression vector carrying the regulatable X P^ promoter, leading to the production of proapolipoprotein A-I at up to 10% of total soluble proteins. The recombinant polypeptide was purified and characterized in terms of apparent molecular mass, isoelectric point, and by both chemical and enzymatic peptide mapping. In addition, it was assayed in vitro for the stimulation of the enzyme lecithin: cholesterol acyltransferase. The data show for the first time that proapo A-I can be produced efficiently in E. coli as a stable and undegraded protein having physical and functional properties indistinguishable from those of the natural product

Trends in language choice and effects of choice on acquisition: Kaqchikel Maya and Spanish in central Guatemala

This study reveals trends in the choice of Kaqchikel Maya, Spanish, or both languages for household activities and by the generations, that is, the children, parents, and grandparents, in a village and a town in the central highlands of Guatemala. Similarities and differences between the village and town are described in both cases. The study also highlights the direct relationship between the language choice of parents and the acquisition of Kaqchikel by children. A total of 171 indigenous residents in the town and the village participated in the study. Findings are based on responses to a sociolinguistic questionnaire that also included measures of knowledge of Kaqchikel. Interviews were conducted by the authors working with bilingual Spanish and Kaqchikel Maya speaking assistants. The measures of knowledge of Kaqchikel were created by the bilingual assistants. The conclusion of this study includes discussion of limitations of current policies for promoting the Kaqchikel language through the education system and broadcasting medi

Effects of Various Pullet Grower Diets on Subsequent Layer Performance

Low protein-low energy, low protein-high energy and high protein-high energy diets were fed to 2100 commercial egg-type pullets for twelve weeks when the pul lets were from 8 to 20 weeks of age. The crude protein and metabolizable energy contents of these diets were 10%, 1950 Gal/kg; 12%, 2900 Gal/kg and 16%, 2900 Gal/kg, respectively

JTT-130, a microsomal triglyceride transfer protein (MTP) inhibitor lowers plasma triglycerides and LDL cholesterol concentrations without increasing hepatic triglycerides in guinea pigs

BACKGROUND: Microsomal transfer protein inhibitors (MTPi) have the potential to be used as a drug to lower plasma lipids, mainly plasma triglycerides (TG). However, studies with animal models have indicated that MTPi treatment results in the accumulation of hepatic TG. The purpose of this study was to evaluate whether JTT-130, a unique MTPi, targeted to the intestine, would effectively reduce plasma lipids without inducing a fatty liver. METHODS: Male guinea pigs (n = 10 per group) were used for this experiment. Initially all guinea pigs were fed a hypercholesterolemic diet containing 0.08 g/100 g dietary cholesterol for 3 wk. After this period, animals were randomly assigned to diets containing 0 (control), 0.0005 or 0.0015 g/100 g of MTPi for 4 wk. A diet containing 0.05 g/100 g of atorvastatin, an HMG-CoA reductase inhibitor was used as the positive control. At the end of the 7(th )week, guinea pigs were sacrificed to assess drug effects on plasma and hepatic lipids, composition of LDL and VLDL, hepatic cholesterol and lipoprotein metabolism. RESULTS: Plasma LDL cholesterol and TG were 25 and 30% lower in guinea pigs treated with MTPi compared to controls (P < 0.05). Atorvastatin had the most pronounced hypolipidemic effects with a 35% reduction in LDL cholesterol and 40% reduction in TG. JTT-130 did not induce hepatic lipid accumulation compared to controls. Cholesteryl ester transfer protein (CETP) activity was reduced in a dose dependent manner by increasing doses of MTPi and guinea pigs treated with atorvastatin had the lowest CETP activity (P < 0.01). In addition the number of molecules of cholesteryl ester in LDL and LDL diameter were lower in guinea pigs treated with atorvastatin. In contrast, hepatic enzymes involved in maintaining cholesterol homeostasis were not affected by drug treatment. CONCLUSION: These results suggest that JTT-130 could have potential clinical applications due to its plasma lipid lowering effects with no alterations in hepatic lipid concentrations

Plant Products for Pharmacology: Application of Enzymes in Their Transformations

Different plant products have been subjected to detailed investigations due to their increasing importance for improving human health. Plants are sources of many groups of natural products, of which large number of new compounds has already displayed their high impact in human medicine. This review deals with the natural products which may be found dissolved in lipid phase (phytosterols, vitamins etc.). Often subsequent convenient transformation of natural products may further improve the pharmacological properties of new potential medicaments based on natural products. To respect basic principles of sustainable and green procedures, enzymes are often employed as efficient natural catalysts in such plant product transformations. Transformations of lipids and other natural products under the conditions of enzyme catalysis show increasing importance in environmentally safe and sustainable production of pharmacologically important compounds. In this review, attention is focused on lipases, efficient and convenient biocatalysts for the enantio- and regioselective formation / hydrolysis of ester bond in a wide variety of both natural and unnatural substrates, including plant products, eg. plant oils and other natural lipid phase compounds. The application of enzymes for preparation of acylglycerols and transformation of other natural products provides big advantage in comparison with employing of conventional chemical methods: Increased selectivity, higher product purity and quality, energy conservation, elimination of heavy metal catalysts, and sustainability of the employed processes, which are catalyzed by enzymes. Two general procedures are used in the transformation of lipid-like natural products: (a) Hydrolysis/alcoholysis of triacylglycerols and (b) esterification of glycerol. The reactions can be performed under conventional conditions or in supercritical fluids/ionic liquids. Enzyme-catalyzed reactions in supercritical fluids combine the advantages of biocatalysts (substrate specificity under mild reaction conditions) and supercritical fluids (high mass-transfer rate, easy separation of reaction products from the solvent, environmental benefits based on excluding organic solvents from the production process)

Supporting information to Influence of data selection on aquatic ecotoxicity characterization factors for selected persistent and mobile substances

&lt;p&gt;Supplementary information to&nbsp;&lt;/p&gt; &lt;h1&gt;Influence of data selection on aquatic ecotoxicity characterization factors for selected persistent and mobile substances&lt;/h1&gt

Trends in language choice and effects of choice on acquisition: Kaqchikel Maya and Spanish in central Guatemala

This study reveals trends in the choice of Kaqchikel Maya, Spanish, or both languages for household activities and by the generations, that is, the children, parents, and grandparents, in a village and a town in the central highlands of Guatemala. Similarities and differences between the village and town are described in both cases. The study also highlights the direct relationship between the language choice of parents and the acquisition of Kaqchikel by children. A total of 171 indigenous residents in the town and the village participated in the study. Findings are based on responses to a sociolinguistic questionnaire that also included measures of knowledge of Kaqchikel. Interviews were conducted by the authors working with bilingual Spanish and Kaqchikel Maya speaking assistants. The measures of knowledge of Kaqchikel were created by the bilingual assistants. The conclusion of this study includes discussion of limitations of current policies for promoting the Kaqchikel language through the education system and broadcasting media.</jats:p