O objetivo neste trabalho foi avaliar a concentração de aminoácidos essenciais (Lisina e Metionina) considerada na literatura como ideal, de acordo com a via de fornecimento (sucedâneo ou concentrado inicial), e sua associação com a suplementação de Glutamato e Glutamina, para bezerros em dois sistemas de aleitamento: convencional ou programado. No primeiro experimento foi realizado o levantamento da composição bromatológica e em aminoácidos dos principais sucedâneos comercializados no Brasil. Nos segundo e terceiro experimentos foram utilizados 45 bezerros da raça Holandês, em delineamento de blocos casualizados distribuídos nos tratamentos: 1) Controle: sem suplementação; 2) Suplementação de lisina e metionina para atingir consumo de 17 e 5,3 g/d, respectivamente, com correção feita com base na análise do sucedâneo, 3) Suplementação de lisina e metionina, para atingir consumo de 17 e 5,3 g/d, respectivamente, com correção feita com base na análise do concentrado inicial. A diferença entre os experimentos foi o sistema de aleitamento ao qual os bezerros foram submetidos: no segundo experimento os animais receberam 6L/d de sucedâneo; enquanto no terceiro estudo os animais foram submetidos ao sistema de aleitamento programado (4L/d até a 2ª semana; 8L/d da 3ª a 6ª semana; 4L/d da 7ª a 8ª semana). No quarto experimento o mesmo delineamento foi utilizado para avaliar, em sistema de aleitamento convencional, os tratamentos: 1) Controle: sem suplementação; 2) Sucedâneo lácteo suplementado com Lisina e Metionina, para atingir consumo de 17 e 5,3g/d, respectivamente + 1% na MS de produto contendo 10% de glutamato e de glutamina; 3) Sucedâneo lácteo suplementado com Lisina e Metionina para atingir consumo de 17 e 5,3g/d + 0,6% na MS de produto contendo 10% de glutamato e de glutamina. Os animais foram alojados em abrigos individuais, com livre acesso a água e concentrado inicial. O consumo de concentrado inicial e o escore fecal dos animais foram registrados diariamente. Semanalmente, foram realizadas pesagens e medidas de crescimento corporal. Nas semanas 2, 4, 6, 8 e 10 de vida foram realizadas colheitas de sangue para determinação de metabólitos como marcadores do status protéico dos animais (albumina, proteína total, N-ureico), status energético (glicose e ?-hidroxibutirato), crescimento ósseo (fosfatase alcalina) e crescimento muscular (creatinina). A composição em aminoácidos dos sucedâneos comercializados no Brasil foi menor que o esperado para dieta que substitui o leite integral. Nos experimentos 2 e 3 a suplementação com lisina e metionina no sucedâneo ou no concentrado inicial para bezerros não resultou em benefícios no desempenho ou no metabolismo. No estudo 4, a suplementação do sucedâneo com lisina, metionina em associação com glutamato e glutamina não alterou o desempenho, a saúde intestinal ou o metabolismo de bezerros leiteiros. / The aim in this work was to evaluate the concentration of essential amino acids (Lysine and Methionine) considered in the literature as ideal, according to feeding route (milk replacer or starter concentrate), and its association with the supplementation of glutamate and glutamine to calves in two feeding systems: conventional or step-down. In the first study, the chemical composition was analyzed and in amino acids of main milk replacer marketed in Brazil. In the second and third studies, 45 Holstein calves were used, in randomized blocks distributed in treatments: 1) Control: without supplementation; 2) Supplementation with lysine and methionine to reach consumption of 17 and 5.3 g/d, respectively, with correction based on the analysis basis of the milk replacer, 3) Supplementation of lysine and methionine to reach consumption of 17 and 5.3 g/d, respectively, with correction based on the analysis basis of starter concentrate. The difference between the experiments was the feeding system applied to the calves: in the second study, the animals received 6 L/d of milk replacer; while in the third study, the animals were submitted to the step-down system (4L/d until the 2nd week; 8L/d of the 3nd to 6th week; 4L/d of the 7th to 8th week). In the fourth study, the same experimental design was used to evaluate, in a conventional feeding system, treatments: 1) Control: without supplementation; 2) AminoGut 0.6%: milk replacer supplemented with Lysine and Methionine, to reach consumption 17 and 5.3g/d, respectively + 0.6% product containing 10% of glutamate and glutamine; 3) AminoGut 1%: milk replacer supplemented with Lysine and Methionine to reach consumption 17 and 5.3g/d + 0.6% product containing 10% of glutamate and glutamine. The animals were housed in individual hutches, with free access to water and starter concentrate. The consumption of starter concentrate and fecal scores of animals were monitored daily. Body growth was weighed and measured weekly. In weeks 2, 4, 6, 8 and 10, blood samples were collected to determine the metabolites as markers of protein status of animals (albumin, total protein, N-urea), energy status (glucose and BHBA), bone growth (alkaline phosphatase) and muscular growth (creatinine). The composition of amino acids of the milk replacer marketed in Brazil was lower than expected for diet that replaces the whole milk. In study 2 and 3, the supplementation of the milk replacer or starter concentrate with lysine and methionine resulted in no benefit on dairy calves performance or metabolism. In study 4 the supplementation of the milk replacer with lysine and methionine in association with glutamate and glutmine had no effect on performance, gut health nor metabolism of dairy calves.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第23055号 / 医博第4682号 / 新制||医||1048(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 妹尾 浩, 教授 伊達 洋至, 教授 長船 健二 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Several papers have reported the enhancing effects of opiate peptides, like Met-enkephalin, on natural killer (NK) cell activity. We examined the actions of Met-enkephalin on NK activity in blood obtained from 18 different donors, of different ages, many of them on several occasions, at several ratios of effector to target cells, several concentrations of peptide, in several types of flasks, with the purity and identity of the pentapeptide verified by chromatography, in a system responsive to interferon, and with results calculated in different ways. No significant increase was found for the peptide for any ratio of cells, any concentration of peptide, or any single subject, even when the subjects with the lowest baseline NK cell activity were used or when the subjects were more than 60 years old. Instead of an increase, the combined results for all subjects at all ratios at all concentrations of Met-enkephalin showed an overall decrease of 31.3 % specific cytotoxicity. These results fail to support the reports of an enhancing effect of Met-enkephalin on NK cell activity.

This study was designed to explore the hormonal regulation of CNS opioid peptide levels in female Sprague Dawley rats. Forty-eight animals were divided into 2 equal groups for acute and chronic studies. Each group was further divided into 4 subgroups, each containing 6 animals. Each rat in the control group received an inert pill (in 0.25 ml corn oil daily by gavage); the second group, 15 μg norethindrone (NE, a potent progestin present in the oral contraceptive Micronor®); the third group, 15 μg NE and 1 μg ethinyl estradiol, EE2 (present in the oral contraceptive Modicon®) and the fourth group, 10 times the dose of the third group. Rats were treated either acutely for 5 days or chronically for 7 weeks. Opioid peptides were estimated by radioimmunoassay. Acute administration of 150 μg NE + 10 μg EE2 decreased the levels of methionine-enkephalin (ME), leucine-enkephalin (LE), dynorphin (DYN) and β-endorphin like immunoreactivity (β-EI) by about 50% in the pituitary. The same dose on chronic administration also decreased DYN, but increased the levels of ME and LE in the pituitary by 331 and 69%, respectively. In the hypothalamus, chronic administration of NE + EE2 increased the level of ME (155%) and LE (87%) as well as of DYN (97%). In the striatum, the levels of LE (33%) and DYN (115%) were elevated during chronic administration. It is concluded that the acute administration of NE + EE2, in general, reduces the levels of ME, LE, DYN and β-EI. The extent of this decrease is about the same in the pituitary, hypothalamus and striatum. Chronic administration of these hormones, however, results in a reversal of this decrease (except for β-EI) and actually can increase the levels of ME, LE and DYN in all three tissues.

Schizophrenia postmortem brain is characterized by γ aminobutyric acid downregulation and by decreased dendritic spine density in frontal cortex. Protracted L-methionine treatment exacerbates schizophrenia symptoms, and our earlier work (Tremolizzo et al. and Dong et al.) has shown that L-methionine decreases reelin and GAD67 transcription in mice which is prevented by co-administration of valproate. In this study, we observed a decrease in spine density following L-methionine treatment, which was prevented by co-administration of valproate. Together with our earlier findings conducted under the same experimental conditions, we suggest that downregulation of spine density in L-methionine-treated mice may be because of the decreased expression of reelin and that valproate may prevent spine downregulation by inhibiting the methylation induced decrease in reelin.

The N-acetyl-L-methionine (NALM) molecule is a methionine (Met) derivative produced via acetylation of the L-Met α-amino group with an N-acetyl group. This molecule has been shown to be bioavailable and capable of fulfilling the dietary requirement for Met in animals and humans. The current experiment was conducted to test a hypothesis that lactating dairy cows fed with NALM would increase milk production by increasing N and energy utilization efficiencies in a dose dependent manner. Eight multiparous Holstein cows that were mid lactation (124 ± 13 days-in-milk) with similar milk production were used in a 4 x 4 Latin square design for 84 d. A developmental NALM product from CJ CheilJedang (Seoul, South Korea) was used as the supplemental source of rumen-protected Met in the present study. Four dietary treatments included 0 g (control), 15 g, 30 g, and 45 g/d/cow of NALM supplementation. Supplementing NALM significantly increased dry matter intake (linear effect; P < 0.01), while milk yield tended to increase quadratically (P = 0.07). A linear decrease in milk fat concentration was seen due to supplementation of NALM in relation to the control ration (P = 0.02). However, milk fat yield was similar across treatments. A trend toward an increase in milk protein yield was observed between the control ration and the ration supplemented with 45 g of NALM (1.18 vs. 1.21 kg/d; P = 0.10). There were no differences in energy-corrected or 3.5% fat-corrected milk yields in response to treatments. It is likely that the supplementation of NALM to mid to late lactating dairy cows may have shifted nutrient and energy utilization toward tissue gain and lactation, which resulted in a decrease in feed efficiency for lactation (P = 0.02). Overall results from the present study suggest that supplementing NALM to mid to late lactating cows can increase milk yield in a dose dependent manner with a shift of net energy partitioning toward milk production and body weight gain. In addition, supplementing NALM increased milk nitrogen (N) output without affecting urinary N excretion.

Rumen protected Methionine (MET) and Lysine (LYS) are critical for milk protein synthesis in dairy cows. N-acetyl-L-methionine (NALM) is a MET derivative that consists of L-Met protected with an acetyl group that is attached to the α-amino group.N-acetyl-L-lysine (NALL) is a LYS derivative that is similarly protected. The objectives of these studies were to quantify the gastrointestinal availability of NALM and NALL. Three experiments were run as 3 × 3 Latin square using 3 second lactation Holstein cows that have been fitted with cannulas in the rumen and duodenum. The cows were fed diets containing the supplements for two weeks prior to each experiment so that the rumen microbes had time to adjust to the supplement. Each period consisted of 10 d of adaptation followed by 2 d of sampling. A dose of 0, 30, or 60 g of NALM was placed under the rumen mat at the time of feeding every day during experiment 1. The cows were similarly supplied with 0, 60, or 120 g of ƐNALL during experiment 2. The cows were supplemented with 0 g, 120 g ofƐNALL, or 120 g of diNALL during experiment 3. On day one of sampling, a liquid marker (Co-EDTA) was also administered at the time of the protected AA administration. Blood, ruminal, and duodenal samples were taken at hours 0, 1, 3, 6, 9, 12, and 24 post-feeding. There were no differences for milk production, milk protein, milk fat, or DMI for NALM or either NALL. There were no differences for ruminal escape (69.1% and 46.2% respectively) and duodenal appearance (2.16% and 3.40% respectively). The ruminal escape of ƐNALL was not different between the 120 g dose (32.7%) and the 60 g dose (27.2%). Duodenal appearance was higher (P < 0.01) for the 60 g dose (2.86%) than for the 120 g dose (1.19%) of ƐNALL. Acetate, propionate, butyrate, and valerate were higher (P < 0.01) for the supplemented cows during experiment 1 with NALL. There were no differences between ƐNALL and diNALL for rumen escape, duodenal appearance, VFA production, or blood LYS AUC. Results of the experiment verify significant protection of the N-acetyl MET and LYS from rumen degradation.

No description available.

Much attention has been recently devoted to sensorially active substances affecting beer quality in the Czech Republic and worldwide. Among them, the heterocyclic and sulphur containing compounds play an important role, some of them with high sensorial activity even in extremely low concentrations. Trace amounts of these compounds, which can be frequently found in foods, participate in formation of their aroma and this effect can be generally evaluated as favorable However, in malt or beer it is true only to a limited extent and the presence of heterocyclic and sulphur containing compounds are in this respect assessed rather unfavorably. The aim of the present study was to provide a survey about of problems in the field of sulphur containing compounds in barley, malt and beer, to describe metabolic paths leading to their formation and to verify experimentally possibilities of their determination using modern analytical methods. Sulphur-containing amino acids are a natural part of barley, malt and beer and are precursors of the origin of volatile sulphur substances. The most frequently occurring sulphur amino acids, metionine, cysteine and homocysteine, were selected for analytical monitoring. The method of gas chromatography was used to determine sulphur-containing amino acids in barley, malt and beer. Prior to the analysis, sulphur-containing amino acids were derived and volatile N(O,S)-ethoxycarbonyl propyl esters were formed; they were subsequently analyzed using the gas chromatography with mass detector (GC/ MSD) and the gas chromatography with flame photo detector (GC/ FPD). Direct analysis of sulphur volatile substances is possible only rarely as they are found in the analyzed matrices (malt, beer) only in very low concentrations ( g/kg,l - ng/kg,l). Before the analysis, the analytes must be extracted from the matrix and concentrated. The modern analytical methods SPME (Solid Phase Micro Extraction), SPDE (Solid Phase Dynamic Extraction) and TDAS (Thermal Desorption Autosampler) were experimentally compared for the extraction and subsequent concentration of sulphur volatile substances. The method of gas chromatography with flame photo detector was used to determine sulphur volatile substances. Following volatile sulphur substances were monitored: dimethyl sulphide, dimethyl disulphide, dimethyl trisulphide, carbon disulphide, ethyl sulphide, diethyl disulphide, methionol, 3-methylthiophen, ethyl thioacetate, 2-methyl-1-buthanthiol. Only metionine was detected in significant amounts in the barley samples analyzed. Not only content but also dependence on a variety and locality were studied. Further, changes in methionine, cysteine and PDMS content during malting were followed. Results proved a significant decline in these substances content depending on the kilning temperature. Three types of fibers were tested for the analyses of the selected volatile sulphur substances in beer in the SPME method. PEG - a fiber with stationary phase Carbowax, PDMS - a fiber with stationary phase polydimethylsiloxan and a combined fiber CAR/PDMS - Carboxen and polydimethylsiloxan. Carbon disulphide, methionol, dimethyl sulphide, 3-methylthiophen and diethyl disulphide were detected with this method. Content of the other analyzed volatile sulphur substances was below the limit of detection. Further was tested usage the SPDE and TDAS methods. Both methods appear to be the suitable for the determination of volatile sulphur substances in beer.

The origin of the 0 -methyl and the N -methyl group in the alkaloid ricinine has been determined. Carbon 14 labelled sodium bicarbonate sodium formate and C14 methyl labelled L-methionine and choline were administered to growing seedlings of Ricinus communis which elaborate the alkaloid ricinine. Methionine was the only metabolite that produced C14 activity in the alkaloid. The activity was located entirely in the methyl groups and the specific activity of the O-methyl and N-methyl group was found, within experimental error, to be identical. / Thesis / Master of Science (MSc)