Substituted glycals as probes of glycosidase mechanisms

D-Glucal and a series of substituted derivatives have been tested as substrates, inhibitors and inactivators of the Agrobacterium faecalis beta-glucosidase in order to probe structure/function relationships in this enzyme. D-Glucal is shown to be a substrate (kcat = 2.3 min-1, Km = 0.85 mM) undergoing hydration with stereospecific protonation from the alpha-face to yield 2-deoxy-beta-D-glucose. 1-Methyl-D-glucal surprisingly serves as only a poor substrate (kcat = 0.056 min-1, Km = 57 mM), also undergoing protonation from the alpha-face. 2-Fluoro-D-glucal, however is completely inert, as a result of inductive destabilisation of the oxocarbenium ion-like transition state for protonation, and functions only as a relatively weak (Ki = 24 mM) inhibitor. Similar behaviour was seen with almond beta-glucosidase and yeast alpha-glucosidase and for the interaction of 2-fluoro-D-galactal with Escherichia coli beta-galactosidase. A series of of alpha, beta-unsaturated glucal derivatives was also synthesised and tested as potential substrates, inhibitors or inactivators of A. faecalis beta-glucosidase. Of these only 1-nitro-D-glucal functioned as a time dependent, irreversible inactivator (ki = 0.011 min-1, Ki = 5.5 mM), presumably acting as a Michael acceptor. Electrospray mass spectrometric analysis revealed multiple labeling of the enzyme by this inactivator, lessening its usefulness as an affinity label. Less reactive Michael acceptor glycals which might have been more specific (1-cyano-, 2-cyano-, 1-carboxylic acid, 1-carboxylic acid methyl ester) unfortunately did not function as inactivators or substrates, only as relatively weak reversible inhibitors (Ki = 3-96 mM).     

The control of protein release from poly(DL-lactide co-glycolide) microparticles by variation of the external aqueous phase surfactant in the water-in oil-in water method

A unique feature of p21 that distinguishes it from the other cyclin-dependent kinase (CDK) inhibitors is its ability to associate with the proliferating cell nuclear antigen (PCNA), an auxiliary factor for DNA polymerases delta and epsilon. While it is now well established that inhibition of cyclin/CDK complexes by p21 can result in G1 cell cycle arrest, the consequences of p21/PCNA interaction on cell cycle progression have not yet been determined. Here, we show, using a tetracycline-regulated system, that expression of wild-type p21 in p53-deficient DLD1 human colon cancer cells inhibits DNA synthesis and causes G1 and G2 cell cycle arrest. Similar effects are observed in cells expressing p21CDK-, a mutant impaired in the interaction with CDKs, but not in cells expressing p21PCNA-, a mutant deficient for the interaction with PCNA. Analysis of cells treated with a p21-derived PCNA-binding peptide provides additional evidence that the growth inhibitory effects of p21 and p21CDK result from their ability to bind to PCNA. Our results suggest that p21 might inhibit cell cycle progression by two independent mechanisms, inhibition of cyclin/CDK complexes, and inhibition of PCNA function resulting in both G1 and G2 arrest.     

Free amino acid supplementation to steers: effects on ruminal fermentation and performance

Three studies were conducted to evaluate amino acid utilization by cattle. In Exp. 1, five steers (580 kg) were fed 86% rolled corn diets with mixtures of amino acids containing up to 6 g/d DL-Met, 24 g/d L-Lys, 6 g/d L-Thr, and 3 g/d L-Trp. Treatments had little effect on ruminal fermentation, diet digestibility, N flow to the duodenum, or microbial efficiency. Ruminal concentrations of Met and Lys increased linearly (P < .05) with amino acid supplementation, whereas Thr responded quadratically, and Trp was not altered. In Exp. 2, four steers (414 kg) were used to measure effects of dietary monensin or laidlomycin propionate in high-grain diets supplemented with amino acids. Ionophores had no significant effect on ruminal fermentation or outflows of amino acids from the rumen. In Exp. 3, 100 steers (287 kg initial BW) were fed diets containing 1% of a nonprotein N source. Treatments were 1) no supplemental N (UREA), 2) UREA plus soybean meal (SBM), 3) UREA plus 2 g/d DL-Met, 8 g/d L-Lys, 2 g/d L-Thr, and 1 g/d L-Trp, or 4) UREA plus 4 g/d DL-Met, 16 g/d L-Lys, 4 g/d L-Thr, and 2 g/d L-Trp. During the growing period (diets based on whole-plant milo silage), gains were higher for SBM-supplemented steers than for UREA steers and intermediate for steers supplemented with amino acids. Few significant differences in performance were observed among treatments during the finishing phase (diets based on dry-rolled corn) or for the entire experiment, but cattle fed SBM or amino acids tended to be fatter and have better marbling scores and quality grades. Amino acids did not greatly alter ruminal fermentation or cattle performance.     

The role of small bowel follow-through examination in the diagnosis of coeliac disease

A retrospective study of 105 barium follow-through examinations on 102 patients suspected of having malabsorption syndrome has been carried out to assess the usefulness of this technique in the diagnosis of coeliac disease. Comparison with histological and biochemical data has been made. Of the 34 examinations on 31 patients with a proven diagnosis of coeliac disease, 30 examinations showed both abnormal X-ray and biopsy findings, two examinations showed normal X-ray appearance but abnormal jejunal biopsy and two showed normal jejunal histology but abnormal X-ray findings. There were two cases with sensitivity, non-invasive nature, ease and cheapness of the technique, barium follow-through examination is suggested as the initial investigation in patients suspected of coeliac disease.     

The Imbrication Model for the integration of social services in a community

An innovative model for organising social services in a community, the Imbrication Model, is contrasted with two traditional models, the Entrepreneurial and the Umbrella Agency. The structural characteristics and dynamics of the three models are illustrated with actual case histories. Imbrication Model calls for the interlocking of personnel from several agencies, with the purpose of redirecting the dysfunctional interagency rivalry prevalent in the traditional models. Imbrications at all organisational levels--Board of Directors, Administrators and Staff--facilitate adoption of the superordinate goal of providing clients with the best services available, regardless of which particular agency delivers the service. Few observers of the current social service scene would challenge the statement that needs for service are unlimited and resources limited. In the USA the imbalance between needs and resources persists despite a decade of massive governmental programmes intended to alleviate social ills. Recent substantial cutbacks in federal funds, moreover are not likely to improve the situation. The resource shortage involves more than a limitation of funds. Deliverers of service and competent programme administrators are also on critically short supply. These shortages are more often than not exacerbated by a chronic spirit of competition among agencies and programmes at the local level. Three organizational models for the delivery and administration of social services, two conventional and one of more recent date, are examined in this article. The innovative model, which has been named the Imbrication Model, explicitly calls for redirecting interagency rivalry and competition. Its ambitious goal is to integrate the efforts of those attempting to meet a community's social service needs.