Performance and tissue zinc and metallothionein accumulation in chicks fed a high dietary level of zinc

Four experiments were conducted to identify several factors that might improve the accuracy and reproducibility of Zn bioavailability assays for chicks. Response of tissue Zn and metallothionein (MT) concentrations to various elevated levels and soluble sources of dietary Zn were measured, as well as the effect of delaying high Zn administration until 7 d posthatching to alleviate the detrimental effect of Zn sulfate on feed intake to 3 wk of age. Bone Zn increased (P < 0.01) in all experiments in response to increasing dietary Zn concentrations. Liver and pancreas MT were affected (P < 0.01) by a source by age interaction and variability that made this criterion unsuitable for bioavailability assays. Lastly, 1-d-old chicks were used to study the effect of delaying feeding of a high-Zn diet up to 7 d of age. The basal diet was fed continuously for 21 d as a control. A diet containing 1,000 ppm Zn was either fed continuously from Day 1, or started on Day 3, 5, or 7. Chicks given high Zn on Day 3, 5, or 7 decreased (P < 0.01) feed intake within 24 h of feeding. Delayed feeding of high dietary Zn might help to alleviate decreased feed intake observed in previous studies. Delaying the onset of high Zn feeding by several days may help alleviate feed intake problems observed with Zn sulfate. Use of either Zn gluconate or Zn acetate as a standard in assays or use of MT synthesis as a bioavailability criterion will probably not be useful to improve accuracy of the estimates.     

The upstream, direct repeat sequence of Prague A Rous sarcoma virus is deficient in mediating efficient Gag assembly and particle release

Rous sarcoma virus (RSV) contains two approximately 135-nt imperfect direct repeats composed of smaller repeats, dr1 (approximately 100 nt) and dr2 (approximately 36 nt), that are between the env and src genes and downstream of src in the 3' untranslated region, respectively. It has previously been shown that a Prague A RSV mutant in which both dr1 sequences are deleted is defective at several points in the virus life cycle, including unspliced RNA and env mRNA stability, unspliced RNA transport, and virus particle assembly. A defect in unspliced RNA transport occurs because a cytoplasmic transport element is present within the dr1. We have suggested that the defect of particle production may arise from the failure of the unspliced RNA to be targeted to sites in the cytoplasm where its translation is favorable for Gag protein assembly. In this report, we have further investigated the function of the direct repeats by comparing virus mutants containing either a single upstream or downstream dr1 sequence. Both mutants were delayed in replication compared to the wild-type; the mutant with a single upstream dr1 (delta DDR) is significantly more defective than the mutant with a single downstream dr1 (delta UDR). While both mutants appear capable of efficiently transporting unspliced RNA to the cytoplasm, the delta DDR mutant with only the upstream dr1 is defective in its ability to support Gag assembly and particle release. The replication defect cannot be repaired by placing the upstream dr1 at the location of the downstream dr1 in the 3' untranslated region. A single point mutation in the upstream dr1 (U to C) restored replication and particle production to near normal levels. The results suggest that unspliced RNA transport and Gag assembly functions may be mediated by different elements within the dr1 and that the Prague A upstream dr1 is defective in the latter but not the former function.     

A dose-dependent increase in mortality with vesnarinone among patients with severe heart failure. Vesnarinone Trial Investigators

BACKGROUND: Vesnarinone, an inotropic drug, was shown in a short-term placebo-controlled trial to improve survival markedly in patients with severe heart failure when given at a dose of 60 mg per day, but there was a trend toward an adverse effect on survival when the dose was 120 mg per day. In a longer-term study, we evaluated the effects of daily doses of 60 mg or 30 mg of vesnarinone, as compared with placebo, on mortality and morbidity. METHODS: We enrolled 3833 patients who had symptoms of New York Heart Association class III or IV heart failure and a left ventricular ejection fraction of 30 percent or less despite optimal treatment. The mean follow-up was 286 days. RESULTS: There were significantly fewer deaths in the placebo group (242 deaths, or 18.9 percent) than in the 60-mg vesnarinone group (292 deaths, or 22.9 percent) and longer survival (P=0.02). The increase in mortality with vesnarinone was attributed to an increase in sudden death, presumed to be due to arrhythmia. The quality of life had improved significantly more in the 60-mg vesnarinone group than in the placebo group at 8 weeks (P<0.001) and 16 weeks (P=0.003) after randomization. Trends in mortality and in measures of the quality of life in the 30-mg vesnarinone group were similar to those in the 60-mg group but not significantly different from those in the placebo group. Agranulocytosis occurred in 1.2 percent of the patients given 60 mg of vesnarinone per day and 0.2 percent of those given 30 mg of vesnarinone. CONCLUSIONS: Vesnarinone is associated with a dose-dependent increase in mortality among patients with severe heart failure, an increase that is probably related to an increase in deaths due to arrhythmia. A short-term benefit in terms of the quality of life raises issues about the appropriate therapeutic goal in treating heart failure.     

Effect of vaccine-induced immune activation on the performance of early-weaned pigs

Two groups of 96 pigs were studied to determine the influence of weaning age, nursery site and a challenge to their immune system on their performance. The weaning ages were 11 to 16 days and 16 to 21 days. One nursery was on-site and the second nursery was off-site. Immune activation was stimulated by the administration of infectious bovine rhinotracheitis virus (IBR) vaccine to half of the pigs at each site. Serum virus neutralisation titres to IBR and total immunoglobulins were monitored in some of the pigs in each group. Performance was measured in terms of feed intake, average daily gain in weight, and feed conversion ratio (FCR). The mean serum immunoglobulin concentrations of all the groups of pigs tended to decrease in the first two weeks after weaning and then increase. Twelve of 20 pigs vaccinated with IBR had neutralisation titres to the virus. The site of the nursery did not significantly affect average daily gain in weight, feed intake or FCR. Pigs weaned at 16 to 21 days of age had a significantly better daily gain in weight than the pigs weaned at 11 to 16 days of age. Immune stimulation of the older weaned pigs did not influence their performance, but it had a significantly (P < 0.016) negative effect on the performance of the younger weaned pigs.     

Empirical models based on free-modulus magnitude estimation of perceived presence in virtual environments

A series of 3 studies was conducted to test free-modulus magnitude estimation as a measure of perceived presence in virtual environments (VEs) and to model the first- and second-order effects of 11 VE system parameters on perceived presence across 5 subtasks. Sequential experimentation techniques were used to build 4 empirical models using polynomial regression. An integrated empirical model of data combined across 2 experiments demonstrated that all significant factors had a positive effect on perceived presence. Three of these parameters--field of view, sound, and head tracking--had almost 3 times as much influence on presence than the other 4 significant parameters, which were visual display resolution, texture mapping, stereopsis, and scene update rate. Sequential experimentation was an efficient tool for building empirical models of perceived presence, but the subjective nature of this phenomenon and individual differences made data bridging across sequential studies problematic. It was concluded that magnitude estimation is a useful measure of perceived presence, and the resulting polynomial regression models can be used to facilitate VE system design decisions. This research has broad application in the selection and design of VE system components and overall design of VE systems.     

Function of galanin in the anterior pituitary of estrogen-treated Fischer 344 rats: autocrine and paracrine regulation of prolactin secretion

Estrogen is a robust stimulator of galanin synthesis and secretion in the anterior pituitary. Galanin is colocalized in lactotrophs in the estrogen-treated anterior pituitary, and its roles in lactotroph function are still being elucidated. In the present studies, we quantified the phenotypes of estrogen-treated Fischer 344 rat anterior pituitary cells expressing the galanin gene by dual in situ hybridization. The total population of galanin-positive pituitary cells increased from undetectable levels to 16% of all cells after 2 weeks of estrogen treatment. More than 90% of the galanin-positive cells coexpressed PRL messenger RNA, and one-third of the lactotrophs expressed galanin messenger RNA. We hypothesized that galanin in the anterior pituitary may contribute to the heterogeneous secretion of PRL, and that one of the functions of galanin is to regulate PRL secretion in an autocrine/paracrine manner. To test this hypothesis, we performed the reverse hemolytic plaque assay combined with in situ hybridization to measure PRL secretion and galanin gene expression within the same individual cells. PRL secretion from galanin-positive lactotrophs was significantly greater than that from galanin-negative lactotrophs. Moreover, treatment with galanin antiserum significantly attenuated PRL secretion from galanin-positive cells, and treatment with galanin significantly enhanced PRL secretion from galanin-negative lactotrophs. In conclusion, these data provide direct evidence that galanin derived from the estrogen-treated anterior pituitary stimulates PRL secretion in both autocrine and paracrine manners.     

Coenzyme A disulfide reductase, the primary low molecular weight disulfide reductase from Staphylococcus aureus. Purification and characterization of the native enzyme

The human pathogen Staphylococcus aureus does not utilize the glutathione thiol/disulfide redox system employed by eukaryotes and many bacteria. Instead, this organism produces CoA as its major low molecular weight thiol. We report the identification and purification of the disulfide reductase component of this thiol/disulfide redox system. Coenzyme A disulfide reductase (CoADR) catalyzes the specific reduction of CoA disulfide by NADPH. CoADR has a pH optimum of 7.5-8.0 and is a dimer of identical subunits of Mr 49,000 each. The visible absorbance spectrum is indicative of a flavoprotein with a lambdamax = 452 nm. The liberated flavin from thermally denatured enzyme was identified as flavin adenine dinucleotide. Steady-state kinetic analysis revealed that CoADR catalyzes the reduction of CoA disulfide by NADPH at pH 7.8 with a Km for NADPH of 2 muM and for CoA disulfide of 11 muM. In addition to CoA disulfide CoADR reduces 4,4'-diphosphopantethine but has no measurable ability to reduce oxidized glutathione, cystine, pantethine, or H2O2. CoADR demonstrates a sequential kinetic mechanism and employs a single active site cysteine residue that forms a stable mixed disulfide with CoA during catalysis. These data suggest that S. aureus employs a thiol/disulfide redox system based on CoA/CoA-disulfide and CoADR, an unorthodox new member of the pyridine nucleotide-disulfide reductase superfamily.