Morphogenetic, structural and production traits of marandu grass under nitrogen rates in Neo soil / [Características morfogênicas, estruturais e produtivas do capim-marandu sob doses de nitrogênio em Neossolo]

Most sandy soils have low natural fertility and low levels of organic matter, making nitrogen (N) fertilization essential. Thus, five doses of N were applied (0, 75, 125, 175 and 225mg dm-³) in a randomized block design to evaluate the effects of nitrogen fertilization on the morphogenetic, structural and production characteristics of Brachiaria brizantha cv. Marandu in a Quartzarenic neosoil. The doses of N did not affect the height of the canopy. The leaf elongation rate, final leaf length and number of live leaves increased linearly at the doses of N. Leaf appearance rate, stem elongation rate, leaf lifespan, phyllochron, leaf senescence rate and tiller density showed a quadratic response to the rates There was also an effect of N rates in herbage mass, leaf mass, stem mass, which increased linearly. Brachiaria brizantha cv. Marandu cultivated in Quartzarenic neosoil requires higher doses of N, 175 and 225mg dm-³. Under these conditions, increases in its morphogenetic, structural and productive characteristics are observed. These findings may not be repeated in the most fertile soils with the greatest capacity to supply N.(AU)

A maioria dos solos arenosos tem baixa fertilidade natural e baixos teores de matéria orgânica, tornando a adubação com nitrogênio (N) essencial. Assim, foram aplicadas cinco doses de N (0, 75, 125, 175 e 225mg dm-³) em delineamento de blocos ao acaso, para se avaliarem os efeitos da adubação com nitrogênio nas características morfogênicas, estruturais e produtivas da Brachiaria brizantha cv. Marandu, em um Neossolo Quartzarênico. As doses de N não afetaram a altura do dossel. A taxa de alongamento foliar, o comprimento final da folha e o número de folhas vivas aumentaram linearmente em função das doses de N. A taxa de aparecimento de folhas, a taxa de alongamento do caule, o tempo de vida da folha, o filocrono, a taxa de senescência foliar e a densidade de perfilhos apresentaram resposta quadrática às doses de N. A massa da forragem, a massa foliar e a massa do caule aumentaram linearmente. Brachiaria brizantha cv. Marandu cultivada em Neossolo Quartzarênico requer maiores doses de N, 175 e 225mg dm-³. Nessas condições, são observados aumentos em suas características morfogenéticas, estruturais e produtivas. Esses achados podem não se repetir nos solos mais férteis e com maior capacidade de suprir N.(AU)

Genetic-Quantitative Study of the First-Service Pregnancy Probability of Murrah Heifers.

Because of the importance of reproduction in stock breeding systems, it is necessary to find selection criteria that increase reproductive efficiency. The aim of this study was to estimate genetic parameters for the probability of conception on first service (PROB) in Murrah heifers, and its association with other traits of economic interest [age at first calving (AFC), service period, calving interval and milk yield at 270 days], with the purpose of evaluating their use as selection criteria. Reproductive information and first lactation records of 1200 Murrah heifers were used to perform two-trait analyses between PROB and the other characteristics. Bayesian inference was used to estimate the variance components, considering PROB as threshold and the other as linear factors. The results demonstrate that this trait has heritability of 0.15, indicating the possibility of a genetic gain by using it for selection. With respect to the genetic correlation estimates, the only high-magnitude association was with AFC (-0.899), which is the current criterion indicating sexual precocity of females. In the light of the parameters estimated, the first-service pregnancy rate is an alternative for indication of sexual precocity, although presenting a smaller genetic gain than the current standard AFC. Nevertheless, additional research should be conducted regarding this trait to assess the economic importance of its use in dairy buffalo production systems.

Molecular recognition: identifying compounds and their targets.

As a result of gene sequencing and proteomic efforts, thousands of new genes and proteins are now available as potential drug targets. The milieu of these proteins is complex and interactive; thousands of proteins activate, inhibit, and control each other's actions. The effect of blocking or activating a protein in a cell is far-reaching, and can affect whole, as well as adjacent pathways. This network of pathways is dynamic and a cellular response can change depending on the stimulus. In this section, the identification and role of individual proteins within the context of networked pathways, and the regulation of the activity of these proteins is discussed. Diverse chemical libraries, combinatorial libraries, natural products, as well as unnatural natural products that are derived from combinatorial biology (Chiu [2001] Proc. Natl. Acad. Sci. USA. 98:8548-8553), provide the chemical diversity in the search for new drugs to block new targets. Identifying new compounds that can become drugs is a long, expensive, and arduous task and potential targets must be carefully defined so as not to waste valuable resources. Equally important is the selection of compounds to be future drug candidates. Target selectivity in no way guarantees clinical efficacy, as the compound must meet pharmaceutical requirements, such as solubility, absorption, tissue distribution, and lack of toxicity. Thus matching biological diversity with chemical diversity involves something more than tight interactions, it involves interactions of the compounds with a variety host factors that can modulate its activity.

Molecular diversity - fifth Lake Tahoe symposium.

This annual conference has been designed to improve communication on chemical and biological molecular diversity research between scientists from industrial and academic sectors. Approximately 150 participants attended, including a mix of physicists, biologists, molecular biologists, chemists, engineers and informaticians. Lake Tahoe, which is surrounded by snow-covered mountains, was a beautiful setting for the conference. Those who enjoy skiing could do so during the afternoon breaks, while others could enjoy walks, visit the local town and take time out for informal scientific discussions.

The ATCG of drug discovery.

The publication of human and microbial genomic sequences has resulted in the availability of large amounts of data that are directly relevant to the discovery of novel and useful drug targets. The elucidation of the function of new genes is not a simple matter and human genetic studies have been among the most useful in identifying genes implicated in disease, although homologs, or orthologs, of human genes in mouse, worm, fly and yeast genomes have also been useful in deciphering gene function. The advent of genomics has led into the study of protein structure and function under the rubric of proteomics. Proteins function in a cell mostly by interacting with other proteins, and protein interaction maps of cellular circuits are now available. Screening strategies to address protein-protein interactions are being developed, and many drug targets in the future are expected to be directed towards these interactions. In addition to using genetic and analytical approaches for finding new drug targets, chemical libraries can be used to inhibit the activity of new proteins, and thus reveal function. The combination of high-throughput screening with testing compounds for ADME (absorption, distribution, metabolism and excretion) and toxicity will help in the early clarification of clinical utility of both new drug targets, as well as drug candidates.

Microbial resistance: novel screens for a contemporary problem.

Historically, natural products have been the source of a large variety of antibacterial agents. In the 1980s, no additional useful antibacterial agents were discovered, leading to the belief that most useful chemotypes from natural product sources had already been discovered. At this time, advances in biotechnology made it feasible to produce sufficient enzyme to set up cell-free screens. Chemical compound libraries and combinatorial synthesis became the source of chemical diversity for the screens. In spite of these efforts, very few new antibacterial agents have been discovered in the last decade. At Small Molecule Therapeutics, Inc., we have developed phenotype-based screens that take advantage of the natural physiology and biochemistry of the target enzymes. We have developed a screen to identify bacterial DNA gyrase and topoisomerase IV poisons. The "hits" identified in this screen are being characterized further. A second screen has also been developed against bacterial topoisomerase 1 in which compounds that cause DNA damage through their interaction with bacterial topoisomerase 1 have been identified. Three of the compounds identified in the screen inhibit DNA relaxation mediated by bacterial topoisomerase 1, induce DNA cleavage, are noncytotoxic at >10 microM, and have MICs of 4.0 microg/mL against Staphylococcus aureus.

Technological advances in high-throughput screening.

A variety of assay technologies continue to be developed for high-throughput screening. These include cell-based assays, surrogate systems using microbial cells such as yeast and bacterial two-hybrid and three-hybrid systems, and systems to measure nucleic acid-protein and receptor-ligand interactions. Modifications have been developed for cell-free, homogeneous assay systems, such as time-resolved fluorescence, fluorescence polarization and the scintillation proximity assay. Innovations in engineering and chemistry have led to delivery systems for nanoliter volumes and sensitive biosensors for ultra-high-throughout screening conducted in nanoliter and picoliter volumes. Spectroscopic methods have been extended to read single molecule fluorescence. Technologies are being developed to identify new targets from genomic information in order to design the next generation of screens.

The effects of space flight on the production of monorden by Humicola fuscoatra WC5157 in solid-state fermentation.

The effect of space flight on the production of the antibiotic monorden on two types of agar media, T8 and PG, by Humicola fuscoatra WC5157 was examined on board the US Space Shuttle mission STS-77 in May 1996. Paired space-flight and ground control samples were prepared using identical hardware, protocol, media, and inoculum. Inoculation occurred simultaneously for both groups 2.5 after launch. The flight and ground samples were allowed to grow for the entire 10-day mission in a dark, thermally controlled (22 degrees C) environment. Post-flight HPLC analysis of the flight and ground sample extracts indicated that the production of monorden by H. fuscoatra WC5157 in the flight samples was higher than in the ground samples in both agar media. In the T8 medium, the production of monorden in the flight and ground samples was 11.6 +/- 3.5 micrograms and 8.9 +/- 1.1 micrograms respectively (30% increase). In the PG medium, the production of monorden in the flight and ground samples was 23.8 +/- 3.3 micrograms and 8.2 +/- 2.2 micrograms respectively (190% increase). The production of monorden in the flight and ground control samples was confirmed by HPLC-MS analysis.

Interaction of NT-4 and BDNF with gp145trkb receptor: effect on cellular metabolism.

In the present study a silicon microphysiometer (Cytosensor) was applied in investigating interactions of gp145(trkb), a member of the tyrosine kinase receptor family, with different neurotrophic factors. NIH-3T3 cells transfected with gp145(trkb) receptors (NIH3T3/trkB cells) were utilized in the studies. Treatment with brain-derived neurotrophic factor (BDNF), neurotrophin-4 (NT-4) and neurotrophin-3 (NT-3) induced changes in the metabolic rate of NIH3T3/trkB cells. In contrast, no response was observed with nerve growth factor (NGF). The effects of NT-4 and BDNF on NIH3T3/trkB cells were receptor-specific in that they did not induce metabolic rate changes in wild type NIH3T3 cells or cells transfected with either gp140(trkb) (TrkA) or gp145(trkb) (TrkC) receptors. In contrast, NT-3 induced metabolic rate changes in cells transfected with each of the three different Trk receptors. The activity of NT-4 was significantly higher than that of BDNF. K252a, a protein kinase inhibitor, reduced the NT-4- and BDNF-induced response of the NIH3T3/trkB cells. This suggests that the NT-4 and BDNF-induced metabolic rate changes are associated with autophosphorylation of the tyrosine protein kinase residues. This hypothesis is further supported by results of western blot analysis. The results show that interactions of Trk receptors with neurotrophic factors result in metabolic changes in cells expressing the receptors.

Melatonin agonists induce phosphoinositide hydrolysis in Xenopus laevis melanophores.

Melatonin, the principal hormone of the vertebrate pineal gland, has been implicated in a variety of neurobiological processes such as circadian rhythmicity and reproductive function. One of the earliest described actions of melatonin was its ability to cause pigment translocation in the dermal melanophores of amphibians. Melatonin binding sites have been identified in the brain of many species and in pigmented tumour cell lines; however, the dermal melanophores of the frog Xenopus Laevis possess the highest known density of melatonin binding sites. These cells are the source from which a melatonin receptor has been cloned and provide an excellent model to study melatonin-mediated signal transduction in an isolated cell system. In Xenopus melanophores, melatonin induces a rapid perinuclear aggregation of intracellular pigment which is associated with a pertussis toxin-sensitive inhibition of cAMP. We have previously demonstrated that a subtype of melatonin binding sites found in selected regions of the pigeon brain and in Syrian Hamster RPMI 1846 melatonin cells are functionally coupled to phosphoinositide hydrolysis as a second messenger. Here we now present evidence to suggest that Xenopus Laevis melanophores also possess melatonin binding sites which are functionally linked to phosphoinositide hydrolysis. Melatonin agonists induced phosphoinositide hydrolysis in melanophores in a concentration-dependent manner with a rank order of potency of 2-iodomelatonin > 6-chloromelatonin > N-acetylserotonin > melatonin. Stimulatory response of 2-iodomelatonin was blocked by the melatonin antagonist N-acetyltryptamine and the alpha-adrenergic antagonist prazosin, which has been shown to have high affinity for melatonin binding sites. Phosphoinositide hydrolysis induced by melatonin agonists was not blocked by the serotonin antagonist ketanserin or by phentolamine, an alpha-adrenergic antagonist, indicating that the response observed was not due to stimulation of 5-HT2a/2c receptors or alpha-adrenergic receptors. Furthermore, incubation of melanophores with the non-hydrolyzable G-protein source GTP-gamma-S attenuated the phosphoinositide dose response induced by 2-iodomelatonin, and pre-incubation of the cells with pertussis toxin had no effect on 2-iodomelatonin-induced phosphoinositide hydrolysis. The present data suggest that Xenopus Laevis Melanophores possess G-protein linked pertussis toxin-insensitive melatonin binding sites which are functionally coupled to phosphoinositide hydrolysis as a signal transduction mechanism.

Ascosteroside, a new antifungal agent from Ascotricha amphitricha. I. Taxonomy, fermentation and biological activities.

Ascosteroside, a novel antifungal compound, was isolated from the culture broth of Ascotricha amphitricha. This compound is an alpha-linked glycoside of a lanostane type triterpenoid. It is active against yeasts such as Candida albicans and Saccharomyces cerevisiae and against filamentous fungi but shows no activity against bacteria. It is not toxic to mammalian cells at concentrations up to 150 microM. In a mouse model, the compound afforded protection comparable to that of ketoconazole.

BMS-192548, a tetracyclic binding inhibitor of neuropeptide Y receptors, from Aspergillus niger WB2346. I. Taxonomy, fermentation, isolation and biological activity.

During the screening of microbial fermentation extracts for their ability to inhibit the binding of 125I-peptid YY (PYY) to the neuropeptide Y (NPY) receptor using the scintillation proximity assay (SPA), BMS-192548 was isolated from the extract of Aspergillus niger WB2346 by bioassay-guided fractionation. BMS-192548 showed the inhibitory activity against 125I-PYY binding to SK-N-MC and SMS-KAN cells, which express NPY1 and NPY2 receptors, respectively, with IC50 values of 24 microM in Y1 and 27 microM in Y2 receptor binding. BMS-192548 demonstrated weak cytotoxicity against murine tumor cell line M-109 with an IC50 value of 240 microM.

Manumycins E, F and G, new members of manumycin class antibiotics, from Streptomyces sp.

Three new manumycin class antibiotics, namely manumycins E, F and G, were isolated from the culture broth of Streptomyces sp. strain WB-8376. Their structures were established by spectroscopic methods, and the S configuration of C-4 in the epoxycyclohexenone moiety was determined by CD exciton chirality method for each of the three compounds. Manumycins E, F and G are active against Gram-positive bacteria, and have moderate inhibitory effects on the farnesylation of p21 ras protein. They demonstrated weak cytotoxic activity against human colon tumor cell HCT-116.

10'-Desmethoxystreptonigrin, a novel analog of streptonigrin.

10'-Desmethoxystreptonigrin, a novel analog of streptonigrin produced by Streptomyces albus, was discovered in a screen for inhibitors of farnesylation of RAS p21 protein. The compound was isolated from the fermentation broth and its structure determined. It is markedly cytotoxic to several human tumor cell lines and also exhibits potent broad-spectrum antibacterial activity.

In vitro and in vivo evaluation of tiacumicins B and C against Clostridium difficile.

Tiacumicins B and C are members of a novel group of 18-membered macrolide antibiotics with in vitro activity against Clostridium difficile. The MICs against 15 strains of C. difficile were 0.12 to 0.25 microgram/ml for tiacumicin B, 0.25 to 1 microgram/ml for tiacumicin C, and 0.5 to 1 microgram/ml for vancomycin. The resistance frequency for both compounds against C. difficile was less than 2.8 x 10(-8) at four and eight times the MIC. The in vivo activities of the tiacumicins against two strains of C. difficile were compared with that of vancomycin in a hamster model of antibiotic-associated colitis. Oral therapy with 0.2, 1, or 5 mg of tiacumicin B or C per kg of body weight protected 100% of clindamycin-treated hamsters exposed to C. difficile ATCC 9689. Oral treatment with identical doses of vancomycin produced a prolonged, dose-dependent survival of hamsters, but it did not prevent the development of fatal colitis at doses of up to 5 mg/kg. When clindamycin-treated animals were exposed to another strain of C. difficile, both tiacumicin B and vancomycin were protective at 5 mg/kg, but not at lower doses. Tiacumicin C was not tested in vivo against the second strain of C. difficile. No tiacumicin B or C was detected in the sera of hamsters treated with single oral doses of 25 mg/kg, while antibiotic levels in the ceca of these hamsters reached 248 micrograms/ml and 285 mg/ml for tiacumicins B and C, respectively. The tiacumicins demonstrated in vitro and in vivo potencies against C. difficile and achieved high concentrations in the cecum, but not the serum, of hamsters after oral administration.