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Trends in population parameters and best linear unbiased prediction of progeny performance in a European F(2) maize population under modified recurrent full-sib selection.

Recurrent selection is a cyclic breeding procedure designed to improve the mean of a population for the trait(s) under selection. Starting from an F(2) population of European flint maize (Zea mays L.) intermated for three generations, we conducted seven cycles of a modified recurrent full-sib (FS) selection scheme. The objectives of our study were to (1) monitor trends across selection cycles in the estimates of the population mean, additive and dominance variances, (2) compare predicted and realized selection responses, and (3) investigate the usefulness of best linear unbiased prediction (BLUP) of progeny performance under the recurrent FS selection scheme applied. Recurrent FS selection was conducted at three locations using a selection rate of 25% for a selection index, based on grain yield and grain moisture. Recombination was performed according to a pseudo-factorial mating scheme, where the selected FS families were divided into an upper-ranking group of parents mated to the lower-ranking group. Variance components were estimated with restricted maximum likelihood. Average grain yield increased 9.1% per cycle, average grain moisture decreased 1.1% per cycle, and the selection index increased 11.2% per cycle. For the three traits we observed, no significant changes in additive and dominance variances occurred, suggesting future selection response at or near current rates of progress. Predictions of FS family performance in Cn+1 based on mean performance of parental FS families in Cn were of equal or higher precision as those based on the mean additive genetic BLUP of their parents, and corresponding correlations were of moderate size only for grain moisture. The significant increase in grain yield combined with the decrease in grain moisture suggest that the F(2) source population with use of a pseudo-factorial mating scheme is an appealing alternative to other types of source materials and random mating schemes commonly used in recurrent selection.

Breeding↗

Transformation of carbon tetrachloride by bisulfide treated goethite, hematite, magnetite, and kaolinite.

This study investigated the transformation of carbon tetrachloride (CT) by goethite, hematite, magnetite, and kaolinite treated with bisulfide to form coatings of iron monosulfide (FeS) and other Fe(II) species. These coatings contribute to abiotic natural attenuation in anaerobic environments. Batch kinetic experiments were performed under anoxic conditions at pH 8.0. Surface-area-normalized pseudo-first-order rate constants for CT transformation did not differ significantly for the three HS- treated iron oxides, but the rate of CT transformation by bisulfide-treated kaolinite was significantly lower, most likely due to kaolinite's lower iron content. The yield of chloroform (CF) from CT transformation was typically approximately 1%. There was negligible or only slight adsorption of several natural organic matter (NOM) model compounds to the surface of HS- treated goethite, and these compounds had no influence on CT transformation rate constants or CF yields. Juglone, on the other hand, adsorbed to a greater extent, and also significantly influenced the CF yield, increasing it by a factor of approximately 20 for HS- treated hematite. We speculate that juglone or its HS- addition product adsorbed to the mineral surface and acted as a hydrogen atom donor that reacted with the trichloromethyl radical intermediate, increasing the CF yield.

Adsorption↗

Application of model-predictive control based on artificial neural networks to optimize the fed-batch process for riboflavin production.

The fed-batch process for commercial production of riboflavin (vitamin B2) was optimized on-line using model-predictive control based on artificial neural networks (ANNs). The information required for process models was extracted from both historical data and heuristic rules. After each cultivation the process model was readapted off-line to include the most recent process data. The control signal (feed rate), however, was optimized on-line at each sampling interval. An optimizer simulated variations in the control signal and assessed the forecasted model outputs according to an objective function. The optimum feed profile for increasing the product yield (YB2/S) and the amount of riboflavin at the time of harvesting was adjusted continuously and applied to the process. In contrast to the control by set-point profiles, the novel ANN-control is able to react on-line to variations in the process and also to incorporate the new process information continuously. As a result, both the total amount of riboflavin produced and the product yield increased systematically by more than 10% and the reproducibility of seven subsequently optimized batches was enhanced.

Algorithms↗

Amplifying the cellular reduction potential of Streptococcus zooepidemicus.

The valuable pharmaceutical polymer, hyaluronic acid, is produced industrially using the gram-positive bacterium Streptococcus zooepidemicus. Synthesis of this polymer is a significant energetic burden upon the microorganism hence the native NADH oxidase gene was cloned and overexpressed to increase the energy yield of catabolism during aerobic cultivation on glucose. Elevated NADH oxidase levels led to a decline in lactic acid generation and prevented ethanol formation, leaving acetate as the main fermentation product. Biomass yield increased due to the energy gained from the formation of acetate. Evaluation of the acetate flux control coefficient over a range of NADH oxidase expression levels revealed that acetate production was sensitive to the NADH oxidase level. However, at high NADH oxidase levels, the acetate flux was mainly influenced by another factor. The concomitant excretion of pyruvate at high NADH oxidase levels suggested that the flux through the pyruvate dehydrogenase enzyme complex was limiting the conversion of pyruvate to acetate.

Acetates↗

The occurrence of salmonellas and lactose-negative Arizonas in reptiles in The Netherlands, and a comparison of three enrichment methods used in their isolation.

A survey was conducted in 1971 in healthy reptiles supplied to the Central Animal Laboratory of Nijmegen for experimental animal research. In order to determine which salmonella serotypes occur, and whether there are several serotypes per animal, several strains of each positive sample were typed.It was found that 160 of 169 samples contained salmonellas or lactose-negative Arizonas or both, and 95 different serotypes were isolated.Of 127 animals examined individually, 67 were carriers of more than one serotype, 42 animals having two types, 21 three types and 4 animals four types.Three enrichment methods were compared. These were tetrathionate broth incubated at 37 degrees C. (T37) and at 43 degrees C. (T43), and selenite broth incubated at 37 degrees C. (SB). All were incubated for 48 hr. before subculture on brilliant-green agar plates. The enrichment methods T37, T43 and SB produced 99, 125 and 123 positive samples respectively, when taken separately. The combinations of T37 and T43, T37 and SB, and T43 and SB produced 145, 142 and 150 positive samples respectively.The yield of serotypes in comparable samples showed no difference between the three enrichment methods. With the use of two methods the yield increased by about 38% compared with one method, and the combination of three methods showed an increase in serotype yield of about 64% compared with one method. A distinct preference by serotypes for definite enrichment methods was not proved.

Animals↗

Accessibility to bacterial nucleic acids of the intercalating drug aliphatic amino acid ellipticinium derivatives in Escherichia coli and Salmonella typhimurium.

The fluorescence of the aliphatic (amino acido)-N2-methyl-9-hydroxyellipticinium (AA-NMHE) derivatives [Auclair, C., Voisin, E., Banoun, H., Bernardou, J., Meunier, B., & Paoletti, C. (1984) J. Med. Chem. 27, 1161-1166], namely, dehydroglycino-NMHE, dehydroalanino-NMHE, dehydrovalino-NMHE, and dehydroleucino-NMHE, has been characterized. The changes in the fluorescence properties of the drugs, including increase in quantum yields, increase in fluorescence lifetimes, and occurrence of energy transfer upon binding to DNA in vitro, have been further investigated. The measurement of the fluorescence increment of AA-NMHE when bound to fluorescent sites inside intact bacteria has been found to be suitable for the determination of the accessibility of the drugs to bacterial nucleic acids according to the method of Lambert and Le Pecq [Lambert, B., & Le Pecq, J.B. (1984) Biochemistry 23, 166-176]. With this methodology, the kinetics of drug uptake, the ability of the drug to reach the bacterial nucleic acids at equilibrium, and the nature of the ligand binding model have been determined in two AA-NMHE-sensitive strains, Escherichia coli BL 101 (Lambert & Le Pecq, 1984) and Salmonella typhimurium TA 98 [Ames, B.N., Lee, F.D., & Durston, W.E. (1973) Proc. Natl. Acad. Sci. U.S.A. 70, 782-786]. The main results obtained are the following: At nonsaturating concentrations, each AA-NMHE exhibits a marked difference in its ability to reach the bacterial nucleic acids. This parameter seems to be correlated with the antibacterial efficiency of the drugs.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaloids↗

Low molecular weight RNA species from chromatin.

Several methods of preparing low molecular weight RNA from chick embryo chromatin have been examined. Traditional methods for dissociating chromatin utilizing high concentrations of salt (greater than 2 M) followed by high-speed centrifugation resulted in very low yields of RNA. Increased yields of RNA were obtained by treating chromatin at lower salt concentration (0.2-0.5 M). By using low salt extraction and sodium dodecyl sulfate-phenol deproteinization, six to eight low molecular weight homogeneous RNA species were isolated from chick embryo chromatin and mouse myeloma chromatin. In the myeloma system, all these RNAs are metabolically stable. Each component is homogeneous as examined by gel electrophoresis and hybridizes with mouse DNA at a rate consistent with a single species. There are multiple gene copies for these RNA species in the mouse genome, varying from 100 to 2000 copies for the different species. One of these RNAs is identical with 5S rRNA. In addition, the redundancy of genes for 18S, 28S, and 5S rRNA and tRNA was determined. Approximately 300 copies for 18 and 28S rTRNA and 500 copies for 5S rRNA were found. tRNAs were on an average 110-fold redundant with about 55 different species measured.

Animals↗

Bubble recycling and ultrasonic cell lysis in a stationary exposure vessel.

An exposure system producing interleafed pulses of 1-MHz ultrasound from balanced, opposing transducers was used to test the bubble recycling hypothesis (as formulated with respect to rotating exposure vessels) in a stationary exposure vessel containing P388 cell suspensions. Cell lysis was measured in cell suspensions which had been exposed or sham-exposed to 1-MHz ultrasound (5 W/cm2 spatial peak pulse average intensity) delivered in continuously alternating, opposing pulses. The opposing pulse cycle frequency (i.e., gating frequency) varied from 0.1 Hz to 100 kHz, corresponding to single pulse durations ranging from 5 microseconds to 5 s. Unequivocal support for the bubble recycling hypothesis was obtained. Lytic yield increased as the gating frequency increased, passing through a maximum at gating frequencies of 100 Hz-1 kHz. As the gating frequency increased further, lytic yield declined rapidly. The results are discussed in relation to recent theoretical estimates of bubble translation speeds in an ultrasound field, and are found to be consistent with those estimates.

Animals↗

Structure-activity relationships for growth inhibition and induction of apoptosis by 4-hydroxy-2-nonenal in raw 264.7 cells.

4-Hydroxy-2-nonenal (HNE) is a highly reactive lipid aldehyde byproduct of the peroxidation of cellular membranes. The structure of HNE features three functional groups, a C1 aldehyde, a C2==C3 double bond, and a C4- hydroxyl group, each of which may contribute to the toxicity of the compound. In addition, the length of the aliphatic chain may influence toxic potency by altering lipophilicity. Using analogous compounds that lacked one or more of the structural moieties, the role of each of these structural motifs in the cytotoxicity of HNE was examined in a mouse alveolar macrophage cell line (RAW 264.7) by a cell survival and growth assay. The importance of these functional groups in the potency of HNE for induction of apoptosis was also examined. The rank order of effects on toxicity was C1---aldehyde >/= C2==C3 double bond >> C4---hydroxyl, with parallel results in both the survival/growth inhibition and apoptosis induction assays. The chain length also influenced toxicity in a series of alpha,beta-unsaturated alkenyl aldehydes, with increasing chain length yielding increasing toxicity. To confirm the importance of the aldehyde moiety, and to examine the role of metabolic detoxification in cellular defenses against HNE toxicity, a RAW 264.7 cell line overexpressing human aldehyde dehydrogenase-3 (hALDH3) was generated. This cell line exhibited nearly complete protection against HNE-protein adduct formation as well as HNE-induced apoptosis. These results illustrate the comparative significance of key structural features of HNE in relation to its potent toxicity and induction of apoptosis.

Aldehydes↗

Silencing of the aspergillopepsin B (pepB) gene of Aspergillus awamori by antisense RNA expression or protease removal by gene disruption results in a large increase in thaumatin production.

Aspergillopepsin B was identified in culture broths of Aspergillus awamori by in situ detection of its proteolytic activity and by immunodetection with anti-aspergillopepsin B antibodies. Severe thaumatin degradation was observed after in vitro treatment of thaumatin with purified aspergillopepsin B. The pepB gene encoding aspergillopepsin B of A. awamori was cloned and characterized. It is located in chromosome IV of A. awamori, as shown by pulsed-field gel electrophoresis, and encodes a protein of 282 amino acids with high similarity to the aspergillopepsin B of Aspergillus niger var. macrosporus. The pepB gene is expressed at high rates as a monocistronic 1.0-kb transcript in media with casein at acidic pH values. An antisense cassette constructed by inserting the pepB gene in the antisense orientation downstream from the gpdA promoter resulted in a good level of antisense mRNA, as shown by reverse transcription-PCR. Partial silencing of the pepB gene by the antisense mRNA resulted in a 31% increase in thaumatin yield. However, significant residual degradation of thaumatin still occurred. To completely remove aspergillopepsin B, the pepB gene was deleted by double crossover. Two of the selected transformants lacked the endogenous pepB gene and did not form aspergillopepsin B. Thaumatin yields increased by between 45% in transformant APB 7/25 and 125% in transformant 7/36 with respect to the parental strain. Reduction of proteolytic degradation by gene silencing with antisense mRNA or total removal of the aspergillopepsin B by directed gene deletion was a very useful method for improving thaumatin production in A. awamori.

Aspergillus↗

Secretion of human serum albumin by Kluyveromyces lactis overexpressing KlPDI1 and KlERO1.

The control of protein conformation during translocation through the endoplasmic reticulum is often a bottleneck for heterologous protein production. The core pathway of the oxidative folding machinery includes two conserved proteins: Pdi1p and Ero1p. We increased the dosage of the genes encoding these proteins in the yeast Kluyveromyces lactis and evaluated the secretion of heterologous proteins. KlERO1, an orthologue of Saccharomyces cerevisiae ERO1, was cloned by functional complementation of the ts phenotype of an Scero1 mutant. The expression of KlERO1 was induced by treatment of the cells with dithiothreitol and by overexpression of human serum albumin (HSA), a disulfide bond-rich protein. Duplication of either PDI1 or ERO1 led to a similar increase in HSA yield. Duplication of both genes accelerated the secretion of HSA and improved cell growth rate and yield. Increasing the dosage of KlERO1 did not affect the production of human interleukin 1beta, a protein that has no disulfide bridges. The results confirm that the ERO1 genes of S. cerevisiae and K. lactis are functionally similar even though portions of their coding sequence are quite different and the phenotypes of mutants overexpressing the genes differ. The marked effects of KlERO1 copy number on the expression of heterologous proteins with a high number of disulfide bridges suggests that control of KlERO1 and KlPDI1 is important for the production of high levels of heterologous proteins of this type.

Endoplasmic Reticulum↗

Pharmacoeconomic consequences of measurement and modification of hospital drug use.

Patterns of drug usage affect hospital-based delivery of healthcare in a variety of ways. Adverse reactions to drugs (ADR) precipitate some 5% of admissions and prejudice the care of some 20% of patients who are in hospital, while inadequate drug therapy prejudices outcomes and prolongs hospital stay. Conversely, appropriate application of drugs can promote recovery and increase the quality of care. Well documented examples include prevention of deep vein thrombosis and postoperative wound infections. Accordingly, optimisation of drug use represents a major quality assurance issue in addition to determining cost-efficiency of healthcare delivery. Drug utilisation review (DUR) requires all elements of the quality assurance process. In practice, therapeutically meaningful and cost-efficient exercises can only be mounted if there is knowledge of the linkage between patterns of drug use and clinical outcomes. These processes of measurement are currently rate-limiting in quality assurance. There are various ways that hospital drug usage can be measured. These range from readily available and relatively cheap quantitative methods to methods requiring the availability of expert staff. There is a sequence of methods involving increasing costs and increasing resource demands yielding increasing detail of information obtained. This sequence commences with pharmacy purchases, followed by pharmacy issues to particular clinical areas, prescription or treatment sheet survey, clinical record review, and finally the reports of trained investigators working in the clinical area. The simpler methods can provide useful information and an efficient basis for choosing and planning definitive studies. Once a category of drug use is appropriately targeted for intervention, drug use can be modified by planned intervention with improvement in clinical outcomes and reduced economic costs in many instances. The intervention strategies to modify drug usage may be classed as re-educative, persuasive, facilitative and power strategies. Other models for implementing behavioural change have been considered, including the impact of trained investigators and the use of online computer prescribing with interactive software with appropriate guidelines. The challenge is to achieve sustained change when interventions are implemented. Cost-efficient quality assurance of drug use is possible with modest resources if outcome-orientated activities are prioritised.

Drug Prescriptions↗

Selection for milk production and persistency using eigenvectors of the random regression coefficient matrix.

The purpose of this study was to investigate the relationships of the eigenvectors of the additive genetic random regression coefficient matrix (K) to selection responses and to determine how many eigenvectors are necessary in the breeding goal to explain the variation. The construction of various eigenvector indexes was based on the K matrix estimated from test-day records of Japanese Holstein cattle. The first (leading) eigenvector index produced constant responses for each day of lactation, indicating that the first eigenvector is responsible for scaling the lactation curve without altering its shape. Daily genetic responses to the second eigenvector index increased linearly as DIM increased. Genetic responses to the third eigenvector index were negative in mid-lactation but were positive in early and late lactation (concave curve). Genetic responses to the fourth and fifth eigenvector indexes hovered around zero across the lactation. The results suggest that both second and third eigenvectors account for the change in the shape of the lactation curve and there is little utility of the fourth and fifth eigenvectors in improving lactation milk or persistency. When the goal is to increase lactation milk yield alone, the index based on the first eigenvector produced a similar response to the index based on all 5 eigenvectors. When the goal is to improve both lactation milk yield and persistency, the index based on the first 3 eigenvectors achieved more than 99.9% of the genetic response to an index based on all 5 eigenvectors. The advantage of an eigenvector index over conventional selection based on total lactation milk yield increases with increasing economic weight assigned to persistency.

Animals↗

Killing and mutation induction in quiescent V79 cells as influenced by nalidixic acid.

Involvement of topoisomerase II in the repair of damage by N-methyl-N'-nitro-N-nitrosoguanidine, UV light and gamma-rays has been studied using quiescent V79 cells. In each case, the presence of nalidixic acid, the inhibitor of topoisomerase II, decreased the survival through suppression of potentially lethal damage repair. There was also an increase in the yield of mutants because of such suppression. The observations were in contrast with those made with exponential cells in the following aspects: (a) density-inhibited cells showed a positive response to suppression of topoisomerase II activity after treatment with UV light and gamma-rays and (b) for MNNG exposure, mutational yield increased instead of decreasing as in exponential cells. The results showed that topoisomerase II played an important part in the repair of damage of density-inhibited cells.

Animals↗

Thrombopoietin therapy increases platelet yields in healthy platelet donors.

The recombinant thrombopoietins have been shown to be effective stimulators of platelet production in cancer patients. It was therefore of interest to determine if one of these, pegylated recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF), could be used to increase platelet counts and consequently platelet yields from apheresis in healthy platelet donors. In a blinded, 2-cycle, crossover study, 59 platelet donors were randomized to receive a single subcutaneous injection of PEG-rHuMGDF (1 microg/kg or 3 microg/kg) or placebo and 15 days later undergo platelet apheresis. Donors treated with placebo had a median peak platelet count after PEG-rHuMGDF injection of 248 x 10(9)/L compared with 366 x 10(9)/L in donors treated with 1 microg/kg PEG-rHuMGDF and 602 x 10(9)/L in donors treated with 3 microg/kg PEG-rHuMGDF. The median maximum percentage that platelet counts increased from baseline was 10% in donors who received placebo compared with 70% in donors who received 1 microg/kg and 167% in donors who received 3 microg/kg PEG-rHuMGDF. There was a direct relationship between the platelet yield and the preapheresis platelet count: Placebo-treated donors provided 3.8 x 10(11) (range 1.3 x 10(11)-7.9 x 10(11)) platelets compared with 5.6 x 10(11) (range 2.6 x 10(11)-12.5 x 10(11)) or 11.0 x 10(11) (range 7.1 x 10(11)-18.3 x 10(11)) in donors treated with 1 microg/kg or 3 microg/kg PEG-rHuMGDF, respectively. Substandard collections (<3 x 10(11) platelets) were obtained from 26%, 4%, and 0% of the placebo, 1 microg/kg, and 3 microg/kg donors, respectively. No serious adverse events were reported; nor were there events that met the criteria for dose-limiting toxicity. Thrombopoietin therapy can increase platelet counts in healthy donors to provide a median 3-fold more apheresis platelets compared with untreated donors.

Adult↗

Ascorbic acid increases the yield of dopaminergic neurons derived from basic fibroblast growth factor expanded mesencephalic precursors.

CNS precursors derived from E12 rat mesencephalon proliferate in the presence of basic fibroblast growth factor and differentiate in vitro into functional dopaminergic neurons, which upon transplantation alleviate behavioral symptoms in a rat model of Parkinson's disease. Here we show that the efficiency of dopaminergic differentiation decreases in the mesencephalic precursors that were proliferated or passaged for extended periods in vitro. Ascorbic acid treatment restored dopaminergic differentiation in these precursors and led to a greater than 10-fold increase in dopamine neuron yield compared with untreated cultures. The effect of ascorbic acid was stereospecific and could not be mimicked by any other antioxidants. The expression of sodium-dependent vitamin C transporter, a recently identified stereospecific ascorbic acid transporter, was maintained in mesencephalic precursors for extended in vitro periods. Pre-treatment of in vitro expanded mesencephalic precursors with ascorbic acid might facilitate the large-scale generation of dopaminergic neurons for clinical transplantation.

Animals↗

UV photolysis of nitrate: effects of natural organic matter and dissolved inorganic carbon and implications for UV water disinfection.

Nitrite (NO2-) formation during ultraviolet (UV) photolysis of nitrate was studied as a function of pH and natural organic matter (NOM) concentration to determine water-quality effects on quantum yields and overall formation potential during UV disinfection of drinking water with polychromatic, medium-pressure (MP) Hg lamps. Quantum yields measured at 228 nm are approximately 2 times higher than at 254 nm under all conditions studied. In the absence of NOM, NO2- quantum yields decrease with time. With addition of NOM, initial quantum yields increase, and the time-dependent decrease is eliminated. At 15 ppm dissolved organic carbon (DOC) as NOM, the quantum yield increases with time. Dissolved inorganic carbon significantly decreases NO2- yields at pH 8 but not pH 6, presumably by reaction of CO2(aq) with peroxynitrite, a major intermediate in NO2- formation. The results indicate important and previously unrecognized roles for NOM and CO2(aq) in nitrate photolysis. When photolysis was carried out using the full spectrum MPUV lamp and germicidally relevant UV doses, NO2- concentrations remained well below the U.S. maximum contaminant level of 1 ppm N, even with nitrate initially present at 10 ppm N. Under current U.S. regulations, NO2- formation should not pose a significant problem for water utilities during UV disinfection of drinking water with MP Hg lamps.

Carbon↗

Association of heifer fertility with cow fertility and yield in dairy cattle.

Genetic correlations of heifer fertility and first-parity yield usually were negative and opposite in sign from genetic correlations of first-parity fertility and yield. Most estimates of genetic correlation between heifer and first-parity fertility were not significantly different from zero. Increasing yield may improve genetic potential for fertility, but stress of increased yield may overcome genetic potential for improved fertility. Days open and 305-day yield for first parity were considered with index selection; sires were evaluated on daughter performance. Days open had a small influence on the selection index; genetic gains in days open for first parity were largely offset by correlated losses in service period for heifers. Restricted index selection, holding days open constant, required considerable economic importance of days open.

Animals↗