Partial purification of tightly bound mitochondrial hexokinase from maize (Zea mays L.) root membranes

In mammals, hexokinase (HK) is strategically located at the outer membrane of mitochondria bound to the porin protein. The mitochondrial HK is a crucial modulator of apoptosis and reactive oxygen species generation. In plants, these properties related to HK are unknown. In order to better understand the physiological role of non-cytosolic hexokinase (NC-HK) in plants, we developed a purification strategy here described. Crude extract of 400 g of maize roots (230 mg protein) contained a specific activity of 0.042 µmol G6P min-1 mg PTN-1. After solubilization with detergent two fractions were obtained by DEAE column chromatography, NC-HK 1 (specific activity = 3.6 µmol G6P min-1 mg PTN-1 and protein recovered = 0.7 mg) and NC-HK 2. A major purification (yield = 500-fold) was obtained after passage of NC-HK 1 through the hydrophobic phenyl-Sepharose column. The total amount of protein and activity recovered were 0.04 and 18%, respectively. The NC-HK 1 binds to the hydrophobic phenyl-Sepharose matrix, as observed for rat brain HK. Mild chymotrypsin digestion did not affect adsorption of NC-HK 1 to the hydrophobic column as it does for rat HK I. In contrast to mammal mitochondrial HK, glucose-6-phosphate, clotrimazole or thiopental did not dissociate NC-HK from maize (Zea mays) or rice (Oryza sativa) mitochondrial membranes. These data show that the interaction between maize or rice NC-HK to mitochondria differs from that reported in mammals, where the mitochondrial enzyme can be displaced by modulators or pharmacological agents known to interfere with the enzyme binding properties with the mitochondrial porin protein.

Russian and Eastern European Dynamic Conditional Correlations in light of the Russian crisis of 2014-2015

This thesis studies the impact of the latest Russian crisis on global markets, and especially Central and Eastern Europe. The results are compared to other shocks and crises over the last twenty years to see how significant they have been. The cointegration process of Central and Eastern European financial markets is also reviewed and updated. Using three separate conditional correlation GARCH models, the latest crisis is not found to have initiated similar surges in conditional correlations to previous crises over the last two decades. Market cointegration for Central and Eastern Europe is found to have stalled somewhat after initial correlation increases post EU accession.

Operant serial feature-positive conditional discrimination with composite features and different topography responses

In a serial feature-positive conditional discrimination procedure the properties of a target stimulus A are defined by the presence or not of a feature stimulus X preceding it. In the present experiment, composite features preceded targets associated with two different topography operant responses (right and left bar pressing); matching and non-matching-to-sample arrangements were also used. Five water-deprived Wistar rats were trained in 6 different trials: X-R®Ar and X-L®Al, in which X and A were same modality visual stimuli and the reinforcement was contingent to pressing either the right (r) or left (l) bar that had the light on during the feature (matching-to-sample); Y-R®Bl and Y-L®Br, in which Y and B were same modality auditory stimuli and the reinforcement was contingent to pressing the bar that had the light off during the feature (non-matching-to-sample); A- and B- alone. After 100 training sessions, the animals were submitted to transfer tests with the targets used plus a new one (auditory click). Average percentages of stimuli with a response were measured. Acquisition occurred completely only for Y-L®Br+; however, complex associations were established along training. Transfer was not complete during the tests since concurrent effects of extinction and response generalization also occurred. Results suggest the use of both simple conditioning and configurational strategies, favoring the most recent theories of conditional discrimination learning. The implications of the use of complex arrangements for discussing these theories are considered.

The Streptococcus mutans GlnR protein exhibits an increased affinity for the glnRA operon promoter when bound to GlnK

The control of nitrogen metabolism in pathogenic Gram-positive bacteria has been studied in a variety of species and is involved with the expression of virulence factors. To date, no data have been reported regarding nitrogen metabolism in the odontopathogenic species Streptococcus mutans. GlnR, which controls nitrogen assimilation in the related bacterial species, Bacillus subtilis, was assessed in S. mutans for its DNA and protein binding activity. Electrophoretic mobility shift assay of the S. mutans GlnR protein indicated that GlnR binds to promoter regions of the glnRA and amtB-glnK operons. Cross-linking and pull-down assays demonstrated that GlnR interacts with GlnK, a signal transduction protein that coordinates the regulation of nitrogen metabolism. Upon formation of this stable complex, GlnK enhances the affinity of GlnR for the glnRA operon promoter. These results support an involvement of GlnR in transcriptional regulation of nitrogen metabolism-related genes and indicate that GlnK relays information regarding ammonium availability to GlnR.

Avaliação da eficiência de dois kits comerciais para detecção de aflatoxina B1 em amostras de milho, ração e amendoim e seus produtos

Foi avaliada a eficiência de dois kits ELISA disponíveis no mercado para detecção de aflatoxina B1, que permitem a triagem de milho contaminado com aflatoxina B1 aos níveis de 5 e 20 ppb. Eles foram comparados ao método de Soares & Rodriguez-Amaya que utiliza cromatografia em camada delgada (CCD) e tem como limite de quantificação 4,0μg/kg (ppb). Foram analisadas 53 amostras incluindo milho, ração para peixe e amendoim e produtos. Os resultados mostraram boa concordância entre as duas técnicas quanto à detecção de aflatoxina B1. O método ELISA mostrou-se mais rápido, menos trabalhoso, utilizando menos solventes orgânicos, porém apresentou resultados presuntivos positivos, tornando necessária a sua confirmação.

Influência da ingestão de biomassas de spirulina (Arthrospira sp.) sobre o peso corporal e consumo de ração em ratos

Neste trabalho verificou-se a influência do consumo de biomassas provenientes de três diferentes origens sobre o peso corporal e consumo de ração em ratos Wistar, machos e adultos. Grupos que consumiram 5% ou 10% (p/p) da biomassa foram comparados com um controle. A biomassa I não promoveu diferença significativa no consumo de ração ou peso corporal dos três grupos. A biomassa II não causou diferença significativa no peso corporal, mas sim no consumo de ração. A biomassa III não causou diferença significativa no consumo de ração, nem no peso corporal, mas houve tendência de maior ganho de peso para o grupo que consumiu a ração contendo 10% de spirulina. Os resultados obtidos indicam que diferentes biomassas podem apresentar diferentes propriedades, mas não confirmam a alegação de que a spirulina pode levar a diminuição de peso ou de consumo de alimento.

Efeito do tempo de fornecimento de ração suplementada com óleo de linhaça sobre a composição físico-química e de ácidos graxos em cabeças de tilápias do Nilo (Oreochromis niloticus)

Este trabalho teve como objetivo realizar um estudo sobre a composição físico-química e, especialmente sobre a composição de ácidos graxos nos lipídios totais de cabeças de tilápias (Oreochromis niloticus), submetidas a diferentes tempos (0, 10, 20, 30 dias) de fornecimento de ração suplementada com óleo de linhaça (fonte do ácido a-linolênico, 18:3n-3 LNA), em substituição ao óleo de girassol. Não houveram diferenças (P>0,05) entre os teores de umidade, cinza, proteína e lipídios totais para os diferentes tratamentos (tempos de fornecimento). Os teores de lipídios totais das cabeças variaram de 8,41 a 10,13% e foi encontrado um total de 27 ácidos graxos nestes lipídios para todos os tratamentos. Os ácidos graxos majoritários foram 18:2n-6 (29,66 a 32,11%), 18:1n-9 (29,68 a 31,71%) e 16:0 (15,53 a 16,63%). Houve um aumento nos teores percentuais do ácido 18:3n-3, da somatória de ácidos graxos ômega-3 e uma diminuição da razão n-6/n-3, com diferenças (P<0,05) entre o tratamento A (tempo zero) e os demais tratamentos que receberam óleo de linhaça. Não houveram diferenças (P>0,05) entre os tratamentos para a razão entre ácidos graxos polinsaturados e saturados (AGPI/AGS).

Relação entre teores de colesterol em filés de tilápias e níveis de óleo de linhaça na ração

O objetivo deste trabalho foi investigar o efeito de rações contendo diferentes níveis de óleo de linhaça no teor de colesterol e na composição centesimal (umidade, cinza, proteína e lipídios totais) de filés de tilápia. A determinação do colesterol foi realizada através de cromatográfica líquida de alta eficiência (CLAE). Esta metodologia foi avaliada no presente trabalho mostrando-se exata e precisa. Os teores de colesterol variaram de 58,3 a 75,5mg/100g de tecido muscular, sendo que a inclusão de óleo de linhaça em um nível de até 3,75% favoreceu a diminuição dos valores de colesterol nos filés de tilápia. No entanto, os níveis de colesterol foram próximos a outras espécies de peixes. Na composição centesimal não houve diferença significativa entre os diferentes tratamentos. Desta forma, a utilização de óleo de linhaça não apresentou influência na composição dos constituintes analisados.

Efeito do fornecimento de ração complementada com semente de linhaça sobre os macronutrientes e colesterol em tecidos de camarões da Malásia (Macrobrachium rosenbergii)

Este estudo teve como objetivo avaliar a composição centesimal dos camarões alimentados com dieta complementada com semente de linhaça. Foram utilizados 800 camarões pós-larvas, divididos em 2 tratamentos: 1) grupo teste à base de caseína complementada com linhaça; e 2) grupo caseína. Os valores médios de umidade (79,8 ± 0,82; 78,9 ± 1,68), cinzas (0,62 ± 0,07; 0,68 ± 0,2), carboidratos (2,63 ± 0,13; 1,58 ± 0,7) e proteínas (15,6 ± 1,02; 16,0 ± 0,58) dos grupos controle e teste, respectivamente, não apresentaram diferença significativa ao final do experimento. Entretanto, a concentração de lipídio total sofreu uma redução significativa (p < 0,05) ao longo do estudo, em ambos os grupos, sendo mais pronunciada no grupo controle (74%), estando ao final do experimento o teor de lipídio mais elevado ( < 0,05) no grupo teste (1,87 ± 0,03) do que no grupo controle (1,12 ± 0,021). Contudo, o teor de colesterol no grupo teste (0,115 ± 0,004) apresentou-se inferior ao do grupo controle (0,1306 ± 0,001), embora sem diferença estatística significativa. Conclui-se que a semente de linhaça na ração parece aumentar o lipídio dos camarões, sugerindo que esse aumento não implica em elevação do colesterol. Havendo, ainda, a possibilidade de ser devido à maior incorporação dos ácidos graxos poliinsaturados.

Efeito da redução da proteína bruta da ração para suínos mantidos em termoneutralidade

Este experimento teve como objetivo avaliar a influência da redução da proteína bruta (PB) e suplementação de aminoácidos sintéticos sobre o desempenho de suínos machos castrados mantidos em ambiente termoneutro (24 °C). Foram utilizados 80 leitões mestiços (Landrace x Large White) com peso médio inicial de 9 kg e idade média de 23 dias. Os animais foram distribuídos em um delineamento inteiramente casualizado, com cinco tratamentos (18, 17, 16, 15 e 14% PB), oito repetições e dois animais por unidade experimental. As rações experimentais foram fornecidas à vontade até o final do experimento, quando os animais atingiram o peso médio de 23 kg (63 dias). A temperatura e a umidade relativa média no interior do galpão foram de 24 °C e 78,7%, respectivamente. O Índice de Temperatura de Globo e Umidade (ITGU) calculado no período foi de 70,4. Não foi observado efeito significativo na redução do nível de proteína bruta da ração sobre as variáveis de desempenho (consumo de ração, ganho de peso e conversão alimentar). Os tratamentos influenciaram os pesos absoluto e relativo do estômago e o peso absoluto do intestino, sendo os maiores valores observados em animais que receberam a ração com maior nível de proteína bruta. Concluiu-se que o nível de PB da ração pode ser reduzido de 18 para 14%, sem prejudicar o desempenho de suínos machos de 9 a 23 kg mantidos em ambiente termoneutro, desde que esta ração seja devidamente suplementada com aminoácidos essenciais limitantes.

Ácidos graxos e composição centesimal da carne de coelhos alimentados com ração contendo farelo de coco

Foi estudado o efeito da inclusão de farelo de coco (FC) na ração de coelhos sobre o perfil de ácidos graxos, a relação de ácidos graxos poli-insaturados para saturados (P/S) e a composição centesimal da carne. O experimento utilizou 60 coelhos, cinco níveis de inclusão de FC na ração (0,00; 6,25; 12,50; 18,75; e 25,00%) e doze repetições por tratamento. Os ácidos palmitoléico, esteárico e linolênico foram afetados linearmente pelo nível de FC na ração. A carne dos animais alimentados sem a adição de FC na ração apresentou maiores (p < 0,05) valores de ácido palmítico em relação aos demais níveis de inclusão; maiores (p < 0,05) níveis dos ácidos mirístico e esteárico e menor (p < 0,05) nível de ácido palmitoléico quando comparada à carne proveniente da ração com 25,00% de FC e menor (p < 0,05) nível de ácido linolênico em relação às carnes provenientes das rações contendo 18,75 e 25,00% de FC. A relação P/S na carne não foi afetada (p > 0,05) pelos tratamentos. O aumento do nível de FC na ração não afetou (p > 0,05) a composição centesimal, indicando que a inclusão desse subproduto na ração de coelhos é viável até 25,00%.

Screening of iron-enriched fungus from natural environment and evaluation of organically bound iron bioavailability in rats

Iron is an essential element for nearly all living organisms, and its deficiency is the most common form of malnutrition in the world. The organic forms of trace elements are considered more bioavailable than the inorganic forms. Although Saccharomyces cerevisiae can enrich metal elements and convert inorganic iron to organic species, its tolerability and transforming capacity are limited. The aim of this study was to screen higher biomass and other iron-enriched fungi strains besides Saccharomyces cerevisiae from the natural environment. A PDA medium containing 800 μg/mL iron was used for initial screening. Fifty strains that tolerated high iron concentration were isolated from the natural environment, and only one strain, No.BY1109, grew well at Fe (II) concentration of 10,000μg/ml. According to morphological characterization, 18S rDNA sequence analysis, and biophysical and biochemical characterization, the strain No.BY1109 was identified as Rhodotorula. The iron content of No.BY1109 (10 mg Fe/g dry cell) was determined using atomic absorption spectrometry. The results of distribution of iron in the cells showed that iron ion was mainly chelated in the cell walls and vacuoles. The bioavailability in rats confirmed that strain No.BY1109 had higher absorption efficiency than that of ferrous sulfate after single dose oral administration. The present study introduces new iron supplements, and it is a basis for finding new iron supplements from natural environment.

盛世事芻蕘Sheng shi chu rao.

Modeste recueil chrétien.Sur la création, la rédemption, l'âme, la rémunération, les fausses doctrines. Par le P. de Mailla. Table générale. Gravé au Ren ai sheng suo.5 sections.

Gaining a better understanding of how Outward Bound Western Canada course outcomes are achieved : a research study /

This study examined how Outward Bound Western Canada (OBWC) course outcomes are achieved by exploring the relationships among course components, students' characteristics, and course outcomes. OBWC is a wilderness-based adventure education organization that helps students achieve outcomes such as increased self-awareness, self-confidence, motivation, interpersonal skills, concern for others, and concern for the environment. This study explored the ways in which the various components of courses and the characteristics of students contribute to determining the outcomes students experience as a result of their courses. The purpose of the study was to gain a better understanding of how OBWC course outcomes are achieved in order to strengthen adventure education theory, enhance practice at OBWC and other adventure education organizations, and provide a foundation for further research on this topic. As an interpretive case study, this study sought to describe how OBWC course outcomes are achieved and to provide interpretations of the research findings. Data was gathered from OBWC students and instructors using the quantitative and qualitative data collection techniques of questionnaire, interview, and observation. Data collected from 98 participants ensured a considerable breadth to the study, while interviews with a number of participants also enabled the collection of in-depth data. Analysis and triangulation of the data from the various sources allowed discernment of the research findings. A comprehensive and detailed picture of how course outcomes are achieved emerged from the findings. Twenty-nine course components were found to influence course outcomes, including various aspects of course activities, the physical environment, instructors, and the group. The findings indicated that certain course components were most influential in determining increases to students' self-awareness, self-confidence, self-reliance, self-esteem, self-concept, motivation, self-responsibility, interpersonal skills, concern for others, and concern for the environment. A number of course components were found to indirectly contribute to positive course outcomes by helping maximize the effectiveness of other components, by increasing students' motivation while on course, or by facilitating the processing and transference of new information. The findings also suggested that several course components either directly or indirectly affected course outcomes in negative ways. In addition, the gender, age, population, and expectations of students were found to play a role in determining the course outcomes they experienced and in determining which course components caused those outcomes. Interpretation of the findings resulted in the generation of research-based theory. The main theoretical argument derived from the results of the study was that course outcomes are influenced by a combination of course components and characteristics of students. More specifically, the theory generated by the study indicated that five groupings of factors contribute to course outcomes, including course activities, the physical environment, instructors, the group, and students' characteristics. The study was considered in relation to existing adventure education literature and larger theoretical issues. The generated theory and research findings were then used to develop suggestions for improving practice at OBWC and other adventure education organizations, as well as for enhancing future research studies.

Enhancing personal effectiveness : the impacts of Costa Rica Outward Bound on students in an independent school in southwestern Ontario

This project examines students in a private school in southwestern Ontario on a 17 -day Costa Rica Outward Bound Rainforest multielement course. The study attempted to discover whether voluntary teenage participants could increase their self-perceptions of life effectiveness by participating in a 17-day expedition. A total of9 students participated in the study. The experimental design that was implemented was a mixed methods design. Participants filled in a Life Effectiveness Questionnaire (LEQ) at four predesignated times during the study. These time intervals occurred (a) before the trip commenced, (b) the first day of the trip, ( c) the last day of the trip, and (d) 1 month after the trip ended. Fieldnotes and recordings from informal group debriefing sessions were also used to gather information. Data collected in this study were analyzed in a variety of ways by the researcher. Analyses that were run on the data included the Friedman test for covariance, means, medians, and the Wilcoxon Pairs Test. The questionnaires were analyzed quantitatively, and the fieldnotes were analyzed qualitatively. Nonparametric statistical analysis was implemented as a result of the small group size of participants. Both sets of data were grouped and discussed according to similarities and differences. The data indicate that voluntary teenage participants experience significant changes over time in the areas of time management, social competency, emotional control, active initiative, and self-confidence. The types of outcomes from this study illustrate that Outward Bound-type opportunities should be offered to teenagers in Ontario schools as a means to bring about self-development.