Dietary L-tryptophan alters aggression in juvenile matrinxa Brycon amazonicus

This study evaluated the effect of dietary supplementation with -tryptophan (L-TRP), a serotonin precursor, on the aggressiveness of juvenile matrinx . Fish were kept in individual aquaria for 7 days receiving the diets: D1 (control: 0.47% of TRP), D2 (0.94% of TRP), D3 (1.88% of TRP), and D4 (3.76% of TRP). After this, they were grouped with an intruder fish to establish a resident-intruder relationship during periods of 20 min. Blood cortisol, glucose, chloride, sodium and calcium; hemoglobin, hematocrit, red blood cell count and volume; liver glycogen and lipids were measured. Territoriality had significant effect on the aggressiveness of matrinx (the residents were more aggressive than intruders, < 0.001) and tryptophan significantly affected their behavior. Fish fed with the D2 diet presented a longer latency until the first attack ( = 0.0069) and bit the intruder fewer times ( = 0.0136) during the period of observation, compared to the control group. The frequency of bites and chases after the first attack was not affected by the dietary supplementation of TRP. Physiological variables were not significantly affected by the diet, except for a moderate increase in cortisol level in fish fed with D2 diet after the fight, indicating slight activation of the hypothalamus-pituitary-interrenal axis. The results show that juvenile matrinx have aggressive and territorial behavior and that a diet containing 9.4 g TRP kg(-1) alter their aggressiveness, without affecting the stress-related physiological parameters.

Taurine bromamine: A potent oxidant of tryptophan residues in albumin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Oxidation of melatonin and its catabolites, N-1-acetyl-N (2)-formyl-5-methoxykynuramine and N-1-acetyl-5-methoxykynuramine, by activated leukocytes

N-1-acetyl-N-2-formyl-5-methoxykynuramine (AFMK) and N-1-acetyl-5-methoxykynuramine (AMK), two melatonin catabolites, have been described as potent antioxidants. We aimed to follow the kinetics of AFMK and AMK formation when melatonin is oxidized by phorbol myristate acetate (PMA) and lipopolysaccharide (LPS)-activated leukocytes. An HPLC-based method was used for AFMK and AMK determination in neutrophil and peripheral blood mononuclear cell cultures supernatants. Samples were separated isocratically on a C18 reverse-phase column using acetonitrile/H2O (25:75) as the mobile phase. AFMK was detected by fluorescence (excitation 340 nm and emission 460 nm) and AMK by UV-VIS absorbance (254 nm). Activation of neutrophils and mononuclear cells with PMA produces larger amounts of AFMK than activation with LPS, probably due to the lower levels of reactive oxygen species formation and myeloperoxidase (MPO) degranulation that occurs when cells are stimulated with LPS. The concentration of AMK found in the supernatant was about 5-10% (from 18-hr cultures) compared with AFMK. This result may reflect its reactivity. Indeed AMK, but not AFMK, is easily oxidized by activated neutrophils in a MPO and hydrogen peroxide-dependent reaction. In conclusion, we defined a simple procedure for the determination of AFMK and AMK in biological samples and demonstrated the capacity of leukocytes to oxidize melatonin and AMK.

Molecular models of tryptophan synthase from Mycobacterium tuberculosis complexed with inhibitors

The development of new therapies against infectious diseases is vital in developing countries. Among infectious diseases, tuberculosis is considered the leading cause of death. A target for development of new drugs is the tryptophan pathway. The last enzyme of this pathway, tryptophan synthase (TRPS), is responsible for conversion of the indole 3-glycerol phosphate into indol and the condensation of this molecule with serine-producing tryptophan. The present work describes the molecular models of TRPS from Mycobacterium tuberculosis (MtTRPS) complexed with six inhibitors, the indole 3-propanol phosphate and five arylthioalkyl-phosphonated analogs of substrate of the a-subunit. The molecular models of MtTRPS present good stereochemistry, and the binding of the inhibitors is favorable. Thus, the generated models can be used in the design of more specific drugs against tuberculosis and other infectious diseases.

Study of the binding of Eu3+ and Tb3+ to L-phenylalanine and L-tryptophan

Trivalent europium and terbium ions have ionic radii similar to that of Ca2+. So they are employed as probes of calcium binding sites in biological molecules. These ions exhibit very useful spectroscopic characteristics, chiefly a pronounced luminescence. In protein bound lanthanide, visible light emission from the lanthanide excited states can be observed when UV light is absorbed by aromatic amino acids. Subsequently, the energy is transferred to the lanthanide ion. The present work was carried out to define the binding sites of Eu3+ and Tb3+ in complexes with the aromatic amino acids L-phenylalanine and L-tryptophan. The techniques utilized were infrared and C nuclear magnetic resonance spectroscopies. It was found that trivalent europium and terbium interact with the carboxylate group of both amino acids. With L-tryptophan, the imino group of the indole ring is also involved representing another coordination site.

Light emission from tryptophan oxidation by hypobromous acid

The emission of ultraweak light from cells is a phenomenon associated with the oxidation of biomolecules by reactive oxygen species. The indole moiety present in tryptophan, serotonin and melatonin is frequently associated with the emission of light during the oxidation of these metabolites. This study presents results for hypobromous acid (HOBr) oxidation of tryptophan as a putative endogenous source of ultraweak light emission. We found that chemiluminescence elicited by the oxidation of tryptophan by HOBr was significantly higher than by hypochlorous acid (HOCl). This difference was related to secondary oxidation reactions, which were more intense using HOBr. The products identified during oxidation by HOCl, but depleted by using HOBr, were N-formylkynurenine, kynurenine, 1,2,3,3a,8,8a-hexahydro-3a-hydroxypyrrolo[2,3-b]-indole-2-carboxylic acid, oxindolylalanine and dioxindolylalanine. The emission of light is dependent on the free α-amino group of tryptophan, and hence, the indole of serotonin and melatonin, although efficiently oxidized, did not produce chemiluminescence. The emission of light was even greater using taurine monobromamine and dibromamine as the oxidant compared to HOBr. A mechanism based on bromine radical intermediates is suggested for the higher efficiency in light emission. Altogether, the experimental evidence described in the present study indicates that the oxidation of free tryptophan or tryptophan residues in proteins is an important source of ultraweak cellular emission of light. This light emission is increased in the presence of taurine, an amino acid present in large amounts in leukocytes, where this putative source of ultraweak light emission is even more relevant.

Digestible tryptophan requirements for broilers from 22 to 42 days old

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The time course of aggressive behaviour in juvenile matrinxa Brycon amazonicus fed with dietary L-tryptophan supplementation

This study evaluated the influence of dietary L-tryptophan (TRP) supplementation on the time course of aggressive behaviour and on neuroendocrine and hormonal indicators in juvenile matrinxA Brycon amazonicus. Supplementation with TRP promoted a change in the fight pattern at the beginning of an interaction with an intruder, resulting in decreased aggressive behaviours during the first 20min. The decrease in aggression did not persist throughout the interaction but increased at 3 and 6h after the beginning of the fight. Monoamine levels in the hypothalamus were not influenced by TRP before or after the fight; however, the hypothalamic serotonin (5-HT) concentration and the 5-hydroxyindole-3-acetic acid (5HIAA):5-HT ratio were significantly correlated with the reduction in aggressive behaviour at the beginning of the fight. Cortisol was not altered by TRP before the fight. After the fight cortisol increased to higher levels in B. amazonicus fed with supplementary TRP. These results indicate that TRP supplementation alters the aggressive behaviour of B. amazonicus and that this effect is limited to the beginning of the fight, suggesting a transient effect of TRP on aggressive behaviour. This is the first study reporting the effects of TRP supplementation on the time course of aggressive interaction in fishes. (C) 2013 The Fisheries Society of the British Isles

Exigências de triptofano e padrão de recuperação do desempenho de poedeiras comerciais após alimentação com rações deficientes em triptofano

Este experimento foi conduzido para avaliar as exigências de triptofano e o padrão de recuperação do desempenho de poedeiras alimentadas com rações deficientes em triptofano. Foram utilizadas 160 poedeiras comerciais da linhagem Hisex White distribuídas em um delineamento inteiramente casualizado com cinco níveis de triptofano nas dietas (0,13; 0,15; 0,17; 0,19 e 0,21%), com oito repetições de quatro aves. As poedeiras permaneceram por duas semanas em adaptação (51 a 52 semanas), por seis semanas para avaliação da exigência de triptofano (53 a 58 semanas) e por quatro semanas para determinação do padrão de recuperação do desempenho (59 a 62 semanas). A produção e a massa de ovos foram prejudicadas quando as poedeiras foram alimentadas com rações contendo 0,13% de triptofano, no entanto, o desempenho foi recuperado após uma semana de alimentação com ração contendo 0,21% desse aminoácido. A qualidade interna dos ovos não foi influenciada pelos níveis de triptofano estudados (ingestão de 137,1 a 228,0 mg triptofano/dia). As exigências de triptofano foram estabelecidas entre 161 e 188 mg/dia, dependendo da característica avaliada (produção ou massa de ovos) e do modelo de regressão aplicado (polinomial, exponencial ou segmentado).

Triptofano na alimentação de codornas japonesas nas fases de recria e postura

Objetivou-se estudar o efeito de diferentes planos alimentares, de acordo com os níveis de triptofano, sobre o desempenho e estresse (pela avaliação de parâmetros fisiológicos) de codornas japonesas (Coturnix coturnix japonica) nas fases de recria e postura. Cento e noventa e duas codornas japonesas na fase de recria (30 a 44 dias de idade) foram alojadas em gaiolas e distribuídas em um delineamento em blocos casualizados, com quatro planos de nutrição [0,27 (controle); 0,52; 0,77 ou 1,02% de triptofano], seis repetições e oito aves por parcela. Foram avaliados os parâmetros de desempenho (consumo de ração, ganho de médio peso, conversão alimentar e taxa de mortalidade) e os parâmetros fisiológicos das aves (relação heterófilo/linfócito e concentração de corticosterona no plasma sangüíneo). Outras 192 codornas japonesas na fase de postura (45 a 146 dias) foram distribuídas em delineamento de blocos ao acaso e submetidas a quatro planos de nutrição (0,23 [controle]; 0,48; 0,73 ou 0,98% de triptofano), com seis repetições e oito aves por parcela. Foram avaliados o consumo de ração, a produção de ovos, a conversão alimentar por dúzia de ovos, a taxa de mortalidade, a relação heterófilo/linfócito e a concentração de corticosterona no plasma. Os níveis de triptofano testados não afetaram o desempenho e os parâmetros fisiológicos de codornas japonesas nas fases de recria e postura.

Effects of oxidation of lysozyme by hypohalous acids and haloamines on enzymatic activity and aggregation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A nonsense mutation in the tyrosinase gene causes albinism in water buffalo

Background: Oculocutaneous albinism (OCA) is an autosomal recessive hereditary pigmentation disorder affecting humans and several other animal species. Oculocutaneous albinism was studied in a herd of Murrah buffalo to determine the clinical presentation and genetic basis of albinism in this species.Results: Clinical examinations and pedigree analysis were performed in an affected herd, and wild-type and OCA tyrosinase mRNA sequences were obtained. The main clinical findings were photophobia and a lack of pigmentation of the hair, skin, horns, hooves, mucosa, and iris. The results of segregation analysis suggest that this disease is acquired through recessive inheritance. In the OCA buffalo, a single-base substitution was detected at nucleotide 1,431 (G to A), which leads to the conversion of tryptophan into a stop codon at residue 477.Conclusion: This premature stop codon produces an inactive protein, which is responsible for the OCA buffalo phenotype. These findings will be useful for future studies of albinism in buffalo and as a possible model to study diseases caused by a premature stop codon.

Digestibilidade aparente do triticale para a tilápia-do-nilo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Relação treonina: lisina para leitões de 6 a 11kg de peso vivo em rações formuladas com base no conceito de proteína ideal

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

LaTBP1: A Leishmania amazonensis DNA-binding protein that associates in vivo with telomeres and GT-rich DNA using a Myb-like domain

Different species of Leishmania can cause a variety of medically important diseases, whose control and treatment are still health problems. Telomere binding proteins (TBPs) have potential as targets for anti-parasitic chemotherapy because of their importance for genome stability and cell viability. Here, we describe LaTBP1 a protein that has a Myb-like DNA-binding domain, a feature shared by most double-stranded telomeric proteins. Binding assays using full-length and truncated LaTBP1 combined with spectroscopy analysis were used to map the boundaries of the Myb-like domain near to the protein only tryptophan residue. The Myb-like domain of LaTBP1 contains a conserved hydrophobic cavity implicated in DNA-binding activity. A hypothetical model helped to visualize that it shares structural homology with domains of other Myb-containing proteins. Competition assays and chromatin immunoprecipitation confirmed the specificity of LaTBP1 for telomeric and GT-rich DNAs, suggesting that LaTBP1 is a new TBP. (C) 2007 Elsevier B.V. All rights reserved.