Redox properties of human medium-chain acyl-CoA dehydrogenase, modulation by charged active-site amino acid residues

The modulation of the electron-transfer properties of human medium-chain acyl-CoA dehydrogenase (hwtMCADH) has been studied using wild-type and site-directed mutants by determining their midpoint potentials at various pH values and estimating the involved pKs. The mutants used were E376D, in which the negative charge is retained; E376Q, in which one negative charge (pKa approximately 6. 0) is removed from the active center; E99G, in which a different negative charge (pKa approximately 7.3) also is affected; and E376H (pKa approximately 9.3) in which a positive charge is present. Em for hwtMCADH at pH 7.6 is -0.114 V. Results for the site-directed mutants indicate that loss of a negative charge in the active site causes a +0.033 V potential shift. This is consistent with the assumption that electrostatic interactions (as in the case of flavodoxins) and specific charges are important in the modulation of the electron-transfer properties of this class of dehydrogenases. Specifically, these charge interactions appear to correlate with the positive Em shift observed upon binding of substrate/product couple to MCADH [Lenn, N. D., Stankovich, M. T., and Liu, H. (1990) Biochemistry 29, 3709-3715], which coincides with a pK increase of Glu376-COOH from approximately 6 to 8-9 [Rudik, I., Ghisla, S., and Thorpe, C. (1998) Biochemistry 37, 8437-8445]. From the pH dependence of the midpoint potentials of hwtMCADH two mechanistically important ionizations are estimated. The pKa value of approximately 6.0 is assigned to the catalytic base, Glu376-COOH, in the oxidized enzyme based on comparison with the pH behavior of the E376H mutant, it thus coincides with the pK value recently estimated [Vock, P., Engst, S., Eder, M., and Ghisla, S. (1998) Biochemistry 37, 1848-1860]. The pKa of approximately 7.1 is assigned to Glu376-COOH in reduced hwtMCADH. Comparable values for these pKas for Glu376-COOH in pig kidney MCADH are pKox = 6.5 and pKred = 7.9. The Em measured for K304E-MCADH (a major mutant resulting in a deficiency syndrome) is essentially identical to that of hwtMCADH, indicating that the disordered enzyme has an intact active site.     

Contingent reinforcement of marijuana abstinence among individuals with serious mental illness: A feasibility study.

The feasibility of using monetary incentives to promote abstinence from marijuana use among individuals with serious mental illness was examined by using a within-subjects experimental design. Participants were 18 adults with schizophrenia or other serious mental illness who reported regular marijuana use. During 2 baseline conditions, participants received payment for submitting urine specimens independent of urinalysis results. During 3 incentive conditions, participants received varying amounts of money if urinalysis results were negative for recent marijuana use. The number of marijuana-negative specimens obtained was significantly greater during incentive than baseline conditions. These results provide evidence that marijuana use among at least some mentally ill individuals is sensitive to contingent reinforcement and support the potential feasibility of using contingency management interventions to reduce substance abuse among the mentally ill. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Features of autoimmune hepatitis in primary sclerosing cholangitis: an evaluation of 114 primary sclerosing cholangitis patients according to a scoring system for the diagnosis of autoimmune hepatitis

Overlapping features between primary sclerosing cholangitis (PSC and autoimmune hepatitis (AIH) have previously been noted. To assess systematically similarities between these disorders, we have evaluated 114 PSC patients (36 women; 78 men), all confirmed by endoscopic retrograde cholangiography (ERC), according to a scoring system proposed by The International Autoimmune Hepatitis Group for the diagnosis of AIH. The scoring system attributes positive or negative scores to the parameters sex, ratio of elevation of serum levels of alkaline phosphatase (ALP) vs. aminotransferase, serum levels of immunoglobulins and autoantibodies, viral markers, history of drug and alcohol intake, genetic factors, liver histology, and response to therapy. Two of the PSC patients (2%) obtained scores above 15 before treatment, satisfying the diagnostic criterion of "definite" AIH. Thirty-eight patients (33%) scored between 10 and 15 points and could be classified as "probable" AIH. The serum level of immunoglobulin G (IgG) was elevated in 68 patients (61% of 111 cases tested), and positive titers of antinuclear antibodies (ANA) or smooth muscle antibodies (SMA) were detected in 24 patients (22% of 111 cases tested). Thirty-five of the PSC patients (33% of 105 evaluable biopsy specimens) obtained positive scores for histological features similar to those of AIH, but the total score for histology was in the negative range in 72 patients (69%) because of the presence of biliary changes. The frequent finding of high scores in PSC patients underlines the similarities PSC may have with AIH. A modification of the scoring system, in particular by increasing the negative score for histological biliary changes, would improve its potential to discriminate between AIH and PSC.     

Adhesion,plasticity and other peculiar properties of solid methane

Adhesion and plasticity of polycrystalline solid methane have been studied in the temperature range 10–90 K. It was shown that adhesion is very strong in the 50–90 K temperature range where the solid methane is soft and sticky. Closer to the melting point (at 90.6 K) the solid methane behaves as extremely viscous non-Newtonian liquid. Below 30 K solid methane loses its stickiness and ductility and acts like ordinary glass. Methane adhesion remains similar for very different materials. We have also made some video observations which reveal unexpected and mostly inexplicable properties of solid methane which could be of some interest for designing new methane moderators for neutron sources and planetary physics particularly the study of the environment of Titan.     

Lake extent changes in Basotu,Tanzania: a mixed-methods approach to understanding the impacts of anthropogenic influence and climate variability

Incompatible land use is a major contributor to ecosystem degradation, and is often exacerbated by climate change impacts. We investigate Lake Basotu, Tanzania as a case study where natural lake variability has been affected by agricultural land use. Comparisons between a satellite-derived history of lake surface area, local precipitation records, and corresponding anthropogenic activity show the impacts of agricultural and historical practices. We argue that insufficient consideration to the wider ecological impacts of large agricultural projects has lasting implications. This is particularly true in semi-arid environments where food production demands need to be continuously met. In the future, major conservation strategies should be investigated to maintain the environmental integrity and sustainability of freshwater resources.     

Effects of Vitamin D3‐Enriched Diet on Egg Yolk Vitamin D3 Content and Yolk Quality

A 40‐wk experiment was conducted using Hy‐Line W‐36 laying hens (19‐wk old) to investigate the impact of feeding cholecalciferol‐enriched diets on egg yolk quality. Feeds were enriched with 4 cholecalciferol levels, 9700 (diet 2), 17200 (diet 3), 24700 (diet 4), and 102200 (diet 5) IU/kg feed. The control (diet 1) contained 2200 IU cholecalciferol/kg feed. Eggs from each replicate group of enriched diets were collected daily and the yolks were pooled into 2‐d period during the first 2 wk. During weeks 3, 4, 6, 8, 12, 16, 20, 24, 28, 32, 36, and 40, pooled samples were generated by daily collection of 3 consecutive days of egg production. The cholecalciferol content of egg yolk from the enriched diets increased rapidly during the first 3 wk. The peak cholecalciferol concentrations in egg yolk that occurred at week 3 were 865, 1641, 2411, and 34815 IU/100 g egg yolk (wet basis) from diet 2 to 5. The average cholecalciferol concentration in yolk during weeks 3 to 40 and the deposition rate of cholecalciferol during the first 3 wk were both linearly increased with the dietary cholecalciferol level when the feed contained no more than 24700 IU/kg cholecalciferol. Egg yolk lipid profile (total lipid content, fatty acid composition, phospholipid composition, and unsaponifiables), physical and functional properties (yolk viscosity and emulsifying property), and sensory quality of hard‐boiled egg yolk were not affected by the cholecalciferol enrichment in the feed. Practical Application: A linear dose‐response relationship between dietary vitamin D₃ level and egg yolk vitamin D₃ content was established at relatively low enrichment levels. Such relationship can be used to formulate feed to achieve a target egg vitamin D level. High vitamin D yolk showed no difference from the conventional yolk in other compositional, functional, and sensory properties.     

Studies of cyclic and linear poly(dimethyl siloxanes): 12. Observation of diffusion behaviour by quasielastic neutron scattering

High resolution neutron scattering experiments have been used to observe the diffusive motion of low molecular weight linear and cyclic poly(dimethyl siloxane) molecules in dilute solution in deuterated benzene. Diffusion coefficients (D) and hydrodynamic radii (R_H) have been compared with values obtained by light scattering for higher molecular weight samples and with radii of gyration (R_g) obtained by small-angle neutron scattering. While the ratio $\text{D}{}_{\mathrm{<mtext>ring</mtext>}}\text{D}{}_{\mathrm{<mtext>chain</mtext>}}$ is close to the predicted value of 0.85, the ratio $\text{R}{}_{\mathrm{g}}\text{R}{}_{\mathrm{<mtext>H</mtext>}}$ falls below the theoretical value for both ring and chain molecules. The scattering curves show effects arising from both centre of mass diffusion and internal molecular motion, and the observed inverse correlation times are compared with calculated behaviour as a function of scattering vector, Q.     

The contribution of electron microscopic cytochemistry to an understanding of the biogenesis of the endoplasmic reticulum of rat hepatocytes

The endoplasmic reticulum has the enzymic machinery for the synthesis of both protein and phospholipid and hence plays a central role in its own biogenesis and that of other cellular membranes. The evidence available concerning the biogenesis of the phospholipid bilayer of the endoplasmic reticulum, particularly from the application of fine structural cytochemical methods for the localization of acyltransferases, is reviewed. The observations are consistent with a model in which phospholipid is synthesized in situ at the site of membrane growth. Synthesis is asymmetric, with most enzymes located at the cytoplasmic side of the membrane, and controlled transmembrane movement of phospholipid results in an asymmetric bilayer.     

Influence of Biodopants on PEDOT Biomaterial Polymers: Using QCM‐D to Characterize Polymer Interactions with Proteins and Living Cells

Organic conducting polymers (OCPs) are currently the subject of intense research in the area of biomaterials and bioelectronics. Of the OCPs, poly(3,4‐ethylenedioxythiophene) (PEDOT) has attracted significant interest, however there has been little work on investigating the incorporation of biological compounds as the dopant species in the polymer which are aimed at enhancing the biocompatibility and biofunctionality of the material. Here, we incorporate the biological dopants dextran sulphate, chondroitin sulphate, and alginate, into PEDOT polymers and investigate their influence on a suite of physicochemical and electrochemical properties. We employ QCM‐D to study the mass of adsorption and the viscoelastic properties of the important extracellular matrix proteins fibronectin and collagen. Furthermore, we use QCM‐D to study the adhesion of PC12 neural cells to the PEDOT‐biodopant polymers with and without an adsorbed protein conditioning layer. QCM‐D was found to be an excellent tool with which to study conducting polymer–biological interactions, with this report the first time that QCM‐D has been used to study cell interactions with conducting polymer biomaterials.     

Long-term outcome after marrow transplantation for severe aplastic anemia

We reviewed the records and reevaluated 212 patients with aplastic anemia transplanted at the Fred Hutchinson Cancer Research Center (FHCRC) between 1970 and 1993 who survived >/=2 years and who have been followed for up to 26 years. Parameters analyzed included hematopoietic function, chronic graft-versus-host disease (GVHD), skin disease, cataracts, lung disease, skeletal problems, posttransplant malignancy, depression, pregnancy/fatherhood, and the return to work or school, as well as patient self-assessment of physical and psychosocial health, social interactions, memory and concentration, and overall severity of symptoms. Survival probabilities at 20 years were 89% for patients without (n = 125) and 69% for patients with chronic GVHD (n = 86) (the status was uncertain in 1 surviving patient). All patients had normal hematopoietic parameters. Skin problems occurred in 14%, cataracts in 12%, lung disease in 24%, and bone and joint problems in 18% of patients. Eleven patients (12%) developed a solid tumor malignancy and 19% of patients experienced depression. Chronic GVHD was the dominant risk factor for late complications. Seventeen patients died at 2.5 to 20.4 years posttransplant; 13 of these had chronic GVHD and related complications. At 2 years, 83% of patients had returned to school or work; the proportion increased to 90% by 20 years. At least half of the patients preserved or regained the ability to become pregnant or father children. Patients rated their quality of life as excellent and symptoms as minimal or mild. In conclusion, marrow transplantation in patients with aplastic anemia established long-term normal hematopoiesis. No new hematologic disorders occurred. The major cause of morbidity and mortality was chronic GVHD. However, the majority of patients who survived beyond 2 years returned to a fully functional life.