Increased plasma acylation-stimulating protein correlates with hyperlipidemia at late gestation

Objectives: Obesity is often associated with negative consequences, including hyperlipidemia and insulin resistance. Weight gain during pregnancy is also associated with major lipid alterations. Fat storage is enhanced in early pregnancy. At late gestation, hyperlipidemia becomes a major manifestation. The acylation‐stimulating protein (ASP) is a potent lipogenic adipocytokine that correlates with postprandial triglyceride (TG) clearance in vivo and has been linked to hyperlipidemic disorders. The role of ASP during a normal pregnancy is unknown. The objective of this study was to investigate plasma ASP levels in correlation with the lipid profile during late gestation. Research Methods and Procedures: Seventy healthy women at late gestation and 60 non‐pregnant controls of similar age and prepregnancy BMI were included in a cross‐sectional study. Fasting plasma ASP levels and the lipid profile of all of the women were measured. Results: ASP levels were markedly elevated in the pregnant women (66%, p < 0.001). ASP levels correlated strongly with the elevated levels of TGs (r = 0.608, p < 0.000), apolipoprotein B (0.519, p < 0.000), and low‐density lipoprotein‐cholesterol (r = 0.405, p < 0.000). Multivariate analysis adjusting for BMI and age showed that changes in ASP levels at late gestation were best predicted by TG and apoB levels, accounting for 53.8% of plasma ASP variation. For the controls, ASP strongly correlated with BMI, which was the only significant predictor of ASP levels. Discussion: Gestational hormone alterations during pregnancy may affect ASP function as a lipogenic factor. Increased plasma ASP levels at late gestation and their strong correlation with parameters reflecting very low‐density lipoprotein accumulation are suggestive of ASP resistance, which may further contribute to the hyperlipidemic state, shifting energy in the form of TGs to the rapidly growing fetus.     

Lignans and secoiridoids from the root bark of Chionanthus virginicus L.: isolation, identification and HPLC analysis

Five lignans, phillyrin (1), pinoresinol-beta-D-glucoside (2), pinoresinol di-beta-D-glucoside (3), phillyrin-2-O-beta-D-glucoside (6), phillyrin-6-O-beta-D-glucoside (7), and three secoiridoids, oleuropein (4), ligustroside (5) and angustifolioside B (8), have been isolated from the root bark of Chionanthus virginicus L. (Oleaceae), a raw material used in the commercial preparation of homeopathy tinctures. Compound 6 is a new lignan, and compounds 2, 3, 4, 5, 7 and 8 are described for the first time in the title plant. The structures of these compounds were elucidated by spectroscopic methods (NMR and HPLC-MS). An isocratic HPLC method for the rapid characterisation and quantification of the major constituents, 1 and 4, of the root bark is described and validated.     

Correlation between mass transfer coefficient k<Subscript>L</Subscript>a and relevant operating parameters in cylindrical disposable shaken bioreactors on a bench-to-pilot scale

Background

Among disposable bioreactor systems, cylindrical orbitally shaken bioreactors show important advantages. They provide a well-defined hydrodynamic flow combined with excellent mixing and oxygen transfer for mammalian and plant cell cultivations. Since there is no known universal correlation between the volumetric mass transfer coefficient for oxygen k_La and relevant operating parameters in such bioreactor systems, the aim of this current study is to experimentally determine a universal k_La correlation.

Results

A Respiration Activity Monitoring System (RAMOS) was used to measure k_La values in cylindrical disposable shaken bioreactors and Buckingham’s π-Theorem was applied to define a dimensionless equation for k_La. In this way, a scale- and volume-independent k_La correlation was developed and validated in bioreactors with volumes from 2 L to 200 L. The final correlation was used to calculate cultivation parameters at different scales to allow a sufficient oxygen supply of tobacco BY-2 cell suspension cultures.

Conclusion

The resulting equation can be universally applied to calculate the mass transfer coefficient for any of seven relevant cultivation parameters such as the reactor diameter, the shaking frequency, the filling volume, the viscosity, the oxygen diffusion coefficient, the gravitational acceleration or the shaking diameter within an accuracy range of +/? 30%. To our knowledge, this is the first k_La correlation that has been defined and validated for the cited bioreactor system on a bench-to-pilot scale.

     

Utility of large consanguineous family-based model for investigating the genetics of type 2 diabetes mellitus

Objectives

This study examined the utility of a family-based model for replicating the results of genome-wide association studies (GWAS) of type 2 diabetes (T2D).

Methods and results

In a total of 232 members of a large consanguineous Omani Arab pedigree (age: 16–80 years), there were 27 diabetics and 50 prediabetics (17 with impaired fasting glucose and 33 with impaired glucose tolerance). All 232 individuals underwent anthropometric and biochemical investigations and genotyped for 14 known common gene variants of modest effect on T2D risk. Power analysis at a LOD score of 3, gave 80% power to locate a single specific locus that accounts for 52% of the total phenotypic variation. Measured genotype analysis (MGA) was used to determine heritability of various quantitative traits (QTs) which ranged 25–56%. Using MGA, some common gene variants were found to have little (< 5%) but significant impact on the heritability of T2D related QTs [KCNJ11 (rs5219), p = 0.004]; [IGF2BP2 (rs4402960), p = 0.02]; [SLC30A8 (rs13266634), p = 0.05]; [CAPN10 (rs2975760), p = 0.031]; [FTO (rs8050136), p = 0.023]; [FTO (rs9939609), p = 0.018] and [SLC30A8 (rs13266634), p = 0.05]. Sib-TDT analysis showed that some gene variants were significantly associated with T2D risk but didn't reach the level of significance after Bonferroni correction [KCNJ11 (rs5219), p = 0.047] and [CAPN10 (rs41266971), p = 0.035].

Conclusion

We have demonstrated that, in principle, a family-based model with minor limitations could be used to replicate some of the results of large GWAS case–control studies. This model could successfully be applied for the future discovery, by deep sequencing, of rare gene variants.     

Functional analysis of cells obtained from bronchoalveolar lavage fluid (BALF) of lung cancer patients

BALF from tumor segments provides access to immune system cells in contact with lung tumors. We analyzed BALF cells as to their production of H2O2 and NO, comparing tumor-affected to non-affected lung segments. Twelve patients were studied (4 NSCLC, 3 SCC, 5 Adenocarcinoma). The cell numbers recovered from BALF varied, and, in adenocarcinoma patients, smaller numbers were recovered from tumor-affected segments. H2O2 production (up to 6.3 nmoles/2x10(5)cells) was obtained in 7/12 patients and, in these, it was more frequent in non-affected segments (7/7) than in affected segments (2/7). After culture, NO production was observed in three patients (6 to 314 microM) that also produced H2O2. These functional characteristics of cells in contact with neoplasia may have a role in determining the fate of the interactions between the immune system and lung cancer.     

Effects of Agricultural Management Policies on the Exposure of Black-Winged Stilts (Himantopus himantopus) Chicks to Cholinesterase-Inhibiting Pesticides in Rice Fields

Levels of exposure to pesticides in rice fields can be significant depending on the environmental policies practiced. The aim of European Union integrated management policy is to reduce pesticide use and impact on environment. Rice fields provide an alternative breeding habitat for many waterbirds that are exposed to the pesticides used and therefore can be valuable indicators of their risk for wildlife. To evaluate integrated management success we examined exposure of Black-winged Stilts (Himantopus himantopus) to cholinesterase-inhibiting pesticides in rice fields under different types of management by measuring plasma cholinesterase activity. Cholinesterase activity was lower in birds sampled in (a) 2008 after a period of intense pesticide application, than in (b) 2005-2007 and 2011 in rice fields subject to integrated management in Doñana (SW Spain) and (c) in control natural wetlands in Spain and Morocco. During 2009 and 2010, cholinesterase activity was lower in rice fields in Doñana than in rice fields in Larache and Sidi Allal Tazi (NW Morocco). Our results suggest that integrated management successfully reduced the exposure of Black-winged Stilts to pesticides in most of the years. Care should be taken to implement mosquito and pest crop controls on time and with environmentally friendly products in order to reduce its impact on wildlife.     

Identification of a glutathione peroxidase inhibitor that reverses resistance to anticancer drugs in human B-cell lymphoma cell lines

Cancer cells isolated from two patients with malignant non-Hodgkin B-cell lymphomas that became resistant to chemotherapy during clinical treatment were made ?fourfold resistant in culture to anticancer drugs, that is cisplatin, etoposide, methotrexate and bortezomib. Because most resistant lines showed significantly increased expression of the anti-oxidative enzyme glutathione peroxidase 1 (GPx1), GPx1 was investigated as a target for inhibitor development. Virtual screening of a library of diverse structures by docking them to the active site of the X-ray crystal structure of bovine GPx1 uncovered compounds that might block the enzyme. An enzyme assay confirmed an acylhydrazone heterocycle (3) with GPx inhibitory activity. Combinations of 3 with the anticancer drugs listed above led to reversal of resistance in the lymphoma cell lines.     

Altered serotonin and dopamine metabolism in the CNS of serotonin 5-HT(1A) or 5-HT(1B) receptor knockout mice

Measurements of serotonin (5-HT), dopamine (DA), and noradrenaline, and of 5-HT and DA metabolites, were obtained by HPLC from 16 brain regions and the spinal cord of 5-HT(1A) or 5-HT(1B) knockout and wild-type mice of the 129/Sv strain. In 5-HT(1A) knockouts, 5-HT concentrations were unchanged throughout, but levels of 5-HT metabolites were higher than those of the wild type in dorsal/medial raphe nuclei, olfactory bulb, substantia nigra, and locus coeruleus. This was taken as an indication of increased 5-HT turnover, reflecting an augmented basal activity of midbrain raphe neurons and consequent increase in their somatodendritic and axon terminal release of 5-HT. It provided a likely explanation for the increased anxious-like behavior observed in 5-HT(1A) knockout mice. Concomitant increases in DA content and/or DA turnover were interpreted as the result of a disinhibition of DA, whereas increases in noradrenaline concentration in some territories of projection of the locus coeruleus could reflect a diminished activity of its neurons. In 5-HT(1B) knockouts, 5-HT concentrations were lower than those of the wild type in nucleus accumbens, locus coeruleus, spinal cord, and probably also several other territories of 5-HT innervation. A decrease in DA, associated with increased DA turnover, was measured in nucleus accumbens. These changes in 5-HT and DA metabolism were consistent with the increased aggressiveness and the supersensitivity to cocaine reported in 5-HT(1B) knockout mice. Thus, markedly different alterations in CNS monoamine metabolism may contribute to the opposite behavioral phenotypes of these two knockouts.