Large scale identification of genes involved in cell surface biosynthesis and architecture in Saccharomyces cerevisiae

The sequenced yeast genome offers a unique resource for the analysis of eukaryotic cell function and enables genome-wide screens for genes involved in cellular processes. We have identified genes involved in cell surface assembly by screening transposon-mutagenized cells for altered sensitivity to calcofluor white, followed by supplementary screens to further characterize mutant phenotypes. The mutated genes were directly retrieved from genomic DNA and then matched uniquely to a gene in the yeast genome database. Eighty-two genes with apparent perturbation of the cell surface were identified, with mutations in 65 of them displaying at least one further cell surface phenotype in addition to their modified sensitivity to calcofluor. Fifty of these genes were previously known, 17 encoded proteins whose function could be anticipated through sequence homology or previously recognized phenotypes and 15 genes had no previously known phenotype.     

Polymer outdoor insulating materials. II. Material considerations

The following article is Part II of a three-part series on the use of polymer insulating materials in outdoor insulation applications. Despite three decades of service history, polymer insulation materials do not have the same level of standardization as porcelain. As with any technology that is not well standardized, misunderstanding and confusion result, which can influence user selection and application. The objective of this series is to review the benefits of polymer insulating materials. Part II discusses the factors that affect the engineering of materials and addresses the validity of some industry perceptions. A future article will explore some of the material science aspects of silicone elastomer materials by using the case of a hybrid insulator design     

Introduction to the AFDC program. Aid to Families with Dependent Children

The NADYA Group, integrated in the Spanish Society of Parenteral and Enteral Nutrition (SENPE), and made up of professionals dedicated to Artificial Nutrition, and specifically, to Artificial Nutrition in the home, annually undertakes the task of collecting data on diagnosis, type of support, follow up characteristics, complications, and quality of life, of patients included in programs of at home artificial nutrition in Spain. In the Annual Register corresponding to 1994, 17 hospitality groups have participated, providing 369 patients with Home Enteral Nutrition, and 30 with Home Parenteral Nutrition. Home Enteral Nutrition is mainly applied in patients with neoplasias (36%) or neurological alterations (35%). The most commonly used access route in the nasogastric tube, although there is an observed increase in the application of Percutaneous Gastrostomies (21%) in relation to previous data of the Spanish population. There is an observed complications index of 0.07 episodes/patient-year, a mortality of 30% (neoplasias) and 20% (neurological alterations), and low rehabilitation indexes in this group. In Home Parenteral Nutrition, post-radiation enteritis, neoplasias, and mesenteric ischemia, are the main diagnostic groups. The majority of the patients have a tunneled tube (63%), with 37% using an implanted tube. With an index of hospitalizations of 0.83 hospitalizations/ patient-year, catheter septicemia justifies the majority of the re-hospitalizations derived from nutritional treatment (0.56 hospitalizations/patient-year), note the mortality of 37%. There are complete rehabilitations, continuing the previously normal activity in 80% of the cases.     

Effect of essential fatty acid depletion on tissue phospholipid fatty acids in spontaneously hypertensive and normotensive rats

Weanling male spontaneously hypertensive (SHR) and normotensive (WKY) rats were maintained on a fat-free semisynthetic diet and killed at various intervals. The effects of fat-depletion on the appearance of essential fatty acid (EFA) deficiency symptoms, the progressive changes of major fatty acids in plasma, liver, heart, and kidney phospholipids (PL), and in skin total lipids were compared between these two strains. After five weeks on the diet, the slower growth and the appearance of EFA deficiency symptoms became evident in SHR. In general, fat-depletion reduced the levels of n−6 fatty acids, whereas it increased those of 20∶3n−9. However, the fat-depletion induced reduction of 18∶2n−6 in heart PL and 20∶4n−6 in kidney, while the elevation of 20∶3n−9 in plasma, heart, and kidney PL were greater in WKY than in SHR. As a result, the elevation of biochemical EFA deficiency index—20∶3n−9/20∶4n−6 ratio—was greater in WKY than in SHR. In comparison with WKY, the concentrations of liver triacylglycerols and the weights of adipose tissues in SHR were reduced to a greater extent, indicating an active catabolism of triacylglycerols in SHR. This study suggests that the earlier appearance of morphological symptoms of EFA deficiency in SHR was not associated with the reducing n−6 EFA levels or with an elevation of triene/tetraene ratio, but possibly to a reduced supply of n−6 EFA for skin prostaglandin synthesis.     

Use of 2'-fluoro-5-methyl-beta-L-arabinofuranosyluracil as a novel antiviral agent for hepatitis B virus and Epstein-Barr virus

A novel anti-hepatitis B virus (anti-HBV) agent, 2'-fluoro-5-methyl-beta-L-arabinofuranosyluracil (L-FMAU), was synthesized and found to be a potent anti-HBV and anti-Epstein-Barr virus agent. Its in vitro potency was evaluated in 2.2.15 and H1 cells for anti-HBV and anti-Epstein-Barr virus activities, respectively. In vitro cytotoxicity in MT2, CEM, 2.2.15, and H1 cells was also assessed, and the results indicated high antiviral selectivities of L-FMAU in these cells.     

Kinetic and Stability Study of an Investigative Antirhinovirus Compound

A hydrolytic acid and base catalyzed ring opening reaction has been demonstrated for a 1,2,4-oxadiazole antiviral compound (WIN 63843) resulting mainly in an amidoxime product. Decomposition products and related impurities were detected using a gradient HPLC method. The hydrolysis reaction was first-order for 35% ethanol/buffer solutions in a 50°C chamber or a light cabinet (1000 ft-candles), the greatest stability being between pH 4 and 6. Furthermore, increasing ethanol concentrations resulted in a great decrease in reaction rates. Therefore, for oral or aerosol solution formulations, light protection, pH control between 4 and 6 and the highest permissible ethanol concentrations would be advantageous. This study has shown that the highly electronegative trifluoromethyl group at the 5 position increases the lability of a 1,2,3-oxdiazole compound.     

Interferon-gamma activation of a mitogen-activated protein kinase, KFR1, in the bloodstream form of Trypanosoma brucei

KFR1, a mitogen-activated protein (MAP) kinase identified in the African trypanosome, Trypanosoma brucei, is a serine protein kinase capable of phosphorylating the serine residues in histone H-1, myelin basic protein, and beta-casein. It phosphorylates four proteins with estimated molecular masses of 22, 34, 46, and 90 kDa from the T. brucei bloodstream-form lysate in vitro. KFR1 bears significant sequence similarity to the yeast MAP kinases KSS1 and FUS3 but cannot functionally complement the kss1/fus3 yeast mutant. It is encoded by a single-copy gene in the diploid T. brucei, and only one of the two alleles can be successfully disrupted, suggesting an essential function of KFR1 in T. brucei. KFR1 activity is present at a much enhanced level in the bloodstream form of T. brucei when compared with that in the insect (procyclic) form. This enhanced activity can be eliminated in vitro by the treatment with protein phosphatase HVH2 known to act specifically on MAP kinases. It can also be decreased in the bloodstream form of T. brucei by serum starvation but induced specifically by interferon-gamma. The production of interferon-gamma in the mammalian host is known to be triggered by T. brucei infection, and this cytokine, as has been reported, promotes the proliferation of T. brucei in the mammalian blood. Since none of these phenomena can be observed in the procyclic form of T. brucei, activation of KFR1 is most likely involved in mediating the interferon-gamma-induced proliferation of T. brucei in the mammalian host.