The mutated forms of the Discs overgrown gene causes overproliferation of imaginal discs of Drosophila melanogaster. Somatic mutations in its human counterpart, casein kinase I epsilon, were strongly associated with human breast cancer. Using the advantage of a high conservancy between fly's dco and human casein kinase I epsilon genes we have chosen D. melanogaster as a model organism to provide a list of probable Dco interaction partners via tandem affinity purification and mass spectrometry analysis. However, these proteins need to be independently verified as true Dco interaction partners.

Estudos recentes indicam que os peptídeos obtidos pela hidrolise enzimática da caseína podem apresentar atividades antioxidantes. Neste trabalho, previamente obteve-se os peptídeos através de hidrolise da caseína utilizando as enzimas Alcalase e Flavourzyme (4h, a 50ºC e pH 8), selecionando os que apresentaram as melhores características, in vitro, relativas à atividade antioxidante. A hidrolise enzimática utilizando a enzima Flavourzyme mostrou melhores resultados, com alto valor de proteína solúvel e conteúdo de aminoácidos livres, além de peptídeos de menor peso molecular do que com a Alcalase, como observado nas análises de cromatografia de permeação em gel e eletroforese em gel de poliacrilamida. Os peptídeos de caseína obtidos com a Flavourzyme também apresentaram melhores resultados utilizando o método ABTS na determinação da capacidade antioxidante. O hidrolisado obtido a partir da enzima Flavourzyme foi aplicado em produtos cárneos e em chocolate branco. Em produtos cárneos, os peptídeos de caseína (2.0%) inibiram, efetivamente, a peroxidação lipídica em carne moída (100%) e em carne mecanicamente separada de ave (CMS) (cerca de 20%) indicando que estes peptídeos podem ser utilizados nestes produtos, auxiliando na prevenção da formação de flavor desagradável e aumentando sua vida útil. Relativamente a sua aplicação em chocolate branco, esta adição teve o intuito de inibir escurecimento deste produto, fator considerado como limitante na sua vida-útil sendo conseqüência tanto de reações de escurecimento não enzimático quanto da oxidação de lipídeos. Os parâmetros que indicaram alteração lipidica e reações não enzimáticas foram mensurados em três diferentes amostras de chocolate branco: uma amostra com 0,2%, de manteiga de cacau, de antioxidante sintético Grindox 562, outra com 0,2%, de manteiga de cacau, dos peptídeos de caseína e a terceira amostra sem qualquer tipo de antioxidante. As amostras foram expostas a duas temperaturas diferentes: 20 ± 2 e 28 ± 2ºC. Os resultados das análises realizadas indicaram que as amostras armazenadas à temperatura de 20ºC apresentaram resultados significativamente melhores àqueles das amostras armazenadas à temperatura de 28ºC, relativos ao índice de acidez, à atividade de água, ao índice de peróxido, à cor e às substâncias reativas ao ácido tiobarbitúrico (TBARS), indicando melhor conservação deste produto. Também foi observado que a adição de quaisquer dos antioxidantes empregados não influenciou de forma significativa os resultados obtidos, evidenciandose assim, que o principal parâmetro responsável pelas alterações do chocolate branco em sua vida útil refere-se à temperatura de armazenamento a qual as amostras foram submetidas. / Recent studies indicate that peptides obtained by casein hydrolysis may have antioxidant activity. In this work, previous casein peptides were obtained by enzymatic hydrolysis using Alcalase and Flavourzyme (4h, at 50ºC and pH 8), selecting the ones that showed the best characteristics in vitro, related to the antioxidant activity. The enzymatic hydrolysis using Flavourzyme showed the best results, with higher soluble protein and free amino acid content and producing lower molecular weight peptides than Alcalase, as observed by gel permeation chromatography and polyacrylamide gel electrophoresis. Related to its application in meat products, casein peptides obtained with Flavourzyme also exhibited greater antioxidant capacity using the ABTS method. The casein hydrolyzed from Flavourzyme enzyme was applicated in ground beef homogenates, mechanically deboned meat (MDM) of poultry and white chocolate. In meat products, casein peptides (2.0%) effectively inhibited lipid peroxidation in ground beef homogenates (100%) and mechanically deboned meat (about 20%) of poultry. Casein peptides may be useful in meat processing as another naturally occurring antioxidant, helping to prevent off-flavor formation in meat products and increasing shelf life. In the use for white chocolate, the goal was to inhibit its browning, the main problems that limit the white chocolate’s shelf-life. Non-enzymatic browning reaction and lipid oxidation were involved directly in the browning of white chocolate. Thus, parameters which indicated fat alteration and non-enzymatic reactions were measured in three different samples of white chocolate. One sample with 0,2% of cocoa butter, with the synthetic antioxidant Grindox 562, other with 0,2% of cocoa butter, with the natural antioxidant and the third sample without any kind of antioxidant. The samples were exposed to two different temperatures: 20 ± 2 and 28 ± 2ºC. The results of the analysis made indicated that the samples stored at the temperature of 20ºC showed results significantly better to those samples stored at the temperature of 28ºC, related to the conservation of the white chocolate. Besides, the results indicated that the addition of any antioxidants employees has not influenced in a significant way the results obtained. Thus, it was evidenced that the main responsible parameter for the alterations of the white chocolate’s shelf-life is related to the storage temperature to which the samples were submitted.

O efeito protetor da caseína fosfopeptídea fosfato de cálcio amorfo (CPP-ACP) contra a erosão dentária é controverso. Este estudo in situ teve como objetivo investigar a capacidade de uma goma de mascar com CPP-ACP em prevenir uma única desmineralização erosiva. Blocos de esmalte bovino (120) selecionados pela dureza superficial inicial foram divididos aleatoriamente entre os grupos: GI - goma de mascar com CPP-ACP, GII - goma de mascar sem CPP-ACP e GIII - controle negativo para avaliação do efeito protetor sem estimulação salivar (sem goma de mascar). Dezenove voluntários participaram do estudo durante três fases cruzadas de 2 h cada. Nas fases de GI e GII os voluntários usaram dispositivos intrabucais palatinos contendo 2 blocos de esmalte, durante 120 minutos e mascaram uma unidade da goma de mascar correspondente ao grupo nos últimos 30 minutos. No grupo controle os voluntários usaram o dispositivo intrabucal por 2h, sem uso de goma de mascar. Em cada fase, imediatamente após a utilização, os dispositivos intrabucais foram imersos em refrigerante tipo cola durante 5 minutos para promover a desmineralização erosiva. A dureza superficial final foi mensurada e os valores foram utilizados para o cálculo do percentual de perda de dureza. Os dados foram analisados por ANOVA de medidas repetidas e teste Tukey (&#x3B1; = 5%). Menor perda de dureza do esmalte foi encontrada após a utilização de goma de mascar com (GI - 32,7%) e sem (GII - 33,5%) CPP-ACP relação ao efeito salivar sem estimulação (GIII - 39,8%) (p <0,05). Não houve diferença entre GI e GII (p> 0,05). Os resultados sugerem que a utilização de goma de mascar imediatamente antes de uma desmineralização erosiva é capaz de diminuir a perda de dureza do esmalte. No entanto, a presença de CPP-ACP na goma de mascar não foi capaz de melhorar este efeito. / The erosion-protective effect of CPP-ACP is controversial. This in situ study aimed to investigate the ability of CPP-ACP chewing gum to prevent a single event of erosive demineralization. Bovine enamel blocks (120), after selection (initial surface hardness) were randomly assigned to groups: GI-chewing gum with CPP-ACP, GIIchewing gum without CPP-ACP and Control group-salivary effect without stimulation (no gum). Nineteen volunteers participated on this study during 3 crossover phases of 2 h. On GI and GII, the volunteers wore intraoral palatal devices for 120 min and chewed a unit of the corresponding chewing gum on the last 30 min. On Control group the volunteer wore the appliance for 2 h, without chewing gum. On each phase immediately after the intraoral use, devices were extra orally immersed in cola drink for 5 minutes to promote erosive demineralization. The percentage of surface hardness loss was calculated. The data were analyzed by Repeated Measures ANOVA and Turkeys test. Less enamel hardness loss was found after the use of chewing gum with (GI-32.7 %) and without (GII-33.5%) CPP-ACP when compared to salivary effect without stimulation (control- 39.8%) (p<0.05). There was no difference between GI and GII (p>0.05). The results suggest that the use of chewing gum immediately before an erosive demineralization is able to diminish the enamel hardness loss. However, the presence of CPP-ACP in the chewing gum cannot enhance this protective effect.

O presente trabalho avaliou o efeito in situ de uma goma de mascar comercialmente disponível contendo caseína fosfopeptídea - fosfato de cálcio amorfo (CPP-ACP) na erosão dentária. Para a primeira etapa do estudo (capacidade remineralizadora) utilizaram-se 72 blocos de esmalte humano selecionados pela dureza de superfície (SHi) e erodidos in vitro pela imersão em Coca Cola®, pH 2,4 por 3 min (avaliação da dureza - SHd). Os blocos foram randomizados entre os grupos: GI Trident Fresh® (sem CPP-ACP), GII controle (sem chiclete) e GIII Trident Total® (com CPPACP). Doze voluntários utilizaram dispositivos intrabucais palatinos por 24 h em 3 fases cruzadas. Nas fases de GI e GIII os voluntários mascaram um chiclete (30 min) e em todas as fases após 2h, a dureza foi avaliada (SHf1). Os blocos foram reposicionados e os dispositivos usados por mais 22 h (+ 3 ciclos de mastigação de chiclete - GI e GIII). A dureza foi reavaliada (SHf2) para cálculo do percentual de recuperação de dureza (%SHR) após 2 e 24h. Na segunda etapa do estudo (ciclagem erosiva) 48 blocos de esmalte humano hígidos foram aleatorizados entre os grupos (GI, GII e GIII) e 8 voluntários utilizaram dispositivos intrabucais palatinos em fases cruzadas de 7 dias cada (washout de 7 dias). O protocolo de ciclagem erosiva foi de 4 imersões diárias do dispositivo intrabucal em 150 ml de Coca Cola® durante 5 min. Nos grupos I e III após cada desafio erosivo e reinserção do dispositivo na cavidade bucal, os voluntários mascaram um chiclete durante 30 min. A alteração da superfície do esmalte foi mensurada por perfilometria (&#x3BC;m). Os dados foram submetidos à ANOVA (2 critérios - etapa 1; 1 critério - etapa 2) e teste Tukey (&#x3B1;=0,05). Os resultados da recuperação de dureza demonstraram haver diferença significativa entre os grupos e os tempos (p<0,05). O Trident Total® (2h = 50,0%; 24h = 95,9%) promoveu maior recuperação de dureza que o Trident Fresh® (2h= 30,0%; 24h= 71,1%) e o grupo controle (2h = 15,7%; 24h = 40,9%). No desafio erosivo prolongado, o Trident Total® (5,2± 2,8 &#x3BC;m) e o Trident Fresh® (3,8 ± 1,5 &#x3BC;m) reduziram significativamente o desgaste dentário em relação ao grupo controle (6,8 ± 3,5 &#x3BC;m), no entanto, não houve diferença significativa entre os chicletes (p>0,05). Conclui-se que o efeito remineralizador da saliva foi maior após utilização de goma de mascar, sendo significativamente aumentado em função do período de remineralização e pela presença de CPP-ACP na goma de mascar. Contudo, para o protocolo de desafio erosivo adotado, apesar da goma de mascar convencional apresentar efeito na inibição do desgaste erosivo, o comportamento da goma de mascar com incorporação de CPP-ACP foi similar. / This study evaluated the in situ effect of a commercial chewing gum containing casein phosphopeptide - amorphous calcium phosphate (CPP-ACP) on dental erosion. On the first stage (remineralizing effect) 72 human enamel blocks, which were selected by surface hardness (SHi) and eroded in vitro by immersion in cola drink, pH 2,4 for 3 minutes (hardness evaluation - SHd) were used. Blocks were randomized into 3 groups: GI Trident Fresh® (conventional gum, without CPP-ACP), GII control (no gum) and GIII Trident Total® (with CPP-ACP). Twelve volunteers wore intraoral palatal devices for 24 h in 3 crossover phases. In phases of GI and GIII volunteers chewed a gum (30 min) and in all phases after 2h, the surface hardness was evaluated (SHf1). The blocks were reinserted and the devices used for additional 22h (+ 3 cycles of chewing gum - GI e GIII). The surface hardness was reassessed (SHf2) to calculate the percentage of surface hardness recovery (%SHR) after 2 and 24h. In the second stage (erosive cycling) 48 healthy human enamel blocks were randomized between groups (GI, GII and GIII) and 8 volunteers wore intraoral palatal devices in crossover phases of 7 days each (washout of 7 days). The cycling protocol consisted of 4 daily immersions of the intraoral device into 150 ml of cola drink for 5 min. In groups I and III after each erosive challenge and oral device reinsertion into oral cavity, the volunteers chewed a gum for 30 min. The enamel surface alterations were measured by profilometry (&#x3BC;m). Data were analyzed by Anova (Two way - stage 1, One way - stage 2) and Turkeys test (&#x3B1;=0,05). The results of percentage of surface hardness recovery showed significant differences for factors groups and time (p<0.05). Trident Total® (2h = 50.0%; 24h = 95.9%) showed higher percentage of surface hardness recovery than the Trident Fresh® (2h = 30.0%; 24h = 71.1%) and control group (2h = 15.7% 24h = 40.9%). In the prolonged erosive challenge, Trident Total® (5.2 ± 2.8 &#x3BC;m) and Trident Fresh® (3.8 ± 1.5 µm) significantly reduced tooth wear compared to control group (6.8 ± 3, 5 µm). However, there was no significant difference between chewing gums. It is concluded that the saliva remineralizing effect increased after the use of conventional chewing gum and was enhanced by prolonged period of remineralization and by the presence of CPPACP in the chewing gum. However for the adopted erosive challenge, despite the conventional chewing gum being able to diminish erosive wear, the chewing gum containing CPP-ACP showed similar effect.

Milch dient der Versorgung neugeborener Säugetiere mit allen lebensnotwendigen Nährstoffen, wie Proteinen, Fetten, Kohlenhydraten, Vitaminen und Mineralstoffen. Insbesondere die Proteingruppe der Caseine, die in der Milch zum Großteil zu Caseinmicellen assoziiert vorliegen, stellen essentielle Aminosäuren bereit und transportieren große Mengen Calcium und Phosphor (Fox, 2008; Horne, 2008). Neben der Bedeutung als erste Nahrungsquelle für Neugeborene haben sich Milch und Milchprodukte zu einem der wichtigsten Handels- und Verarbeitungsgüter entwickelt. In Deutschland ist vorrangig der Konsum von Kuhmilch verbreitet. Zunehmend wird aber auch die Milch von Büffel, Schaf und Ziege direkt oder verarbeitet konsumiert (Fox, 2008; Töpel, 2016). Dabei unterscheidet sich die Zusammensetzung der Milch zwischen den verschiedenen Tierarten zum Teil beträchtlich. Diese Unterschiede betreffen nicht nur die Anteile der Hauptnährstoffe, sondern auch die Eigenschaften und die Zusammensetzung der Caseinmicellen (CM) (Fox, 2008). Die Eigenschaften und die Stabilität gegenüber einer Calciumkomplexierung wurden für bovine Caseinmicellen in der Literatur bereits ausführlich diskutiert (Horne, 1984; Banon & Hardy, 1992; Needs et al., 2000; O’Connell et al., 2001; Huppertz et al., 2004). Über die Micellen anderer Tierarten lagen zu Beginn der Untersuchungen hingegen kaum Informationen vor. Ein Ziel der vorliegenden Arbeit war es deshalb, die Caseinmicellen aus der Milch der wiederkäuenden Paarhufer Kuh, Büffel, Schaf, Ziege und Kamel und der nicht wiederkäuenden Unpaarhufer Pferd und Esel sowie aus Humanmilch bezüglich Größe, Struktur, Zusammensetzung und Stabilität zu charakterisieren und daraus Erkenntnisse zum tierartspezifischen Micellaufbau abzuleiten. Hierzu wurden die Caseinmicellen mit Hilfe der Ultrazentrifugation von der Molke abgetrennt und in tierartspezifischem, synthetischem Milchultrafiltrate (SMUF) resuspendiert. Untersuchungen der hydrodynamischen Durchmesser mittels dynamischer Lichtstreuung zeigten, dass die CM der Nicht-Wiederkäuer Stute und Esel sowie des Kamels deutlich größer waren als die der Wiederkäuer Kuh, Büffel, Schaf und Ziege, wohingegen sich die humanen CM als die kleinsten zeigten. Mittels Rasterelektronenmikroskopie (REM) konnten aufgrund der beobachteten unterschiedlichen Oberflächenstrukturen für einige Tierarten Hinweise hinsichtlich einer größen- und κ-Casein-abhängigen Ausbildung von κ Casein ‚bunches‘ oder einer ‚hairy layer‘ an der Oberfläche der kolloidalen Partikel erhalten werden. Anhand der Aminosäurezusammensetzung der micellaren Proteine konnte abgeleitet werden, dass bei allen Tierarten vor allem Phosphoserin sowie die hydrophoben Aminosäuren maßgeblich am Aufbau der CM beteiligt sein dürften. Hierbei wurden jedoch tierartspezifische Unterschiede festgestellt. Bei den Wiederkäuern wurde eine besonders starke Assoziation der Caseinmonomere über Phosphoserincluster vermutet, während bei den Nicht-Wiederkäuern und insbesondere bei den humanen CM auch hydrophobe Wechselwirkungen eine entscheidende Rolle spielen. Calcium und anorganischer Phosphor konnten als die bedeutendsten micellaren Salze identifiziert werden, die in besonders hohen Gehalten in den CM von Stute und Esel vorkamen. Humane Micellen wiesen deutlich niedrigere Mineralstoffgehalte auf, weshalb die Bedeutung der elektrostatischen Wechselwirkungen für die Verknüpfung der Caseinmonomere geringer eingeschätzt wurde als bei den weiteren untersuchten Tierarten. In Bezug auf die Stabilität gegenüber einer Calciumkomplexierung mit Ethylenglycol-bis(2-aminoethylether)-N,N,N\',N\'-tetraessigsäure (EGTA) unterschieden sich die CM der Tiere sehr deutlich voneinander. Instabile Micellen zeichneten sich durch eine starke Abnahme der Trübung, eine Zunahme des extra-micellaren Caseins sowie eine schneller einsetzende Verkleinerung des hydrodynamischen Durchmessers aus. Für die Micellstabilität ergab sich die folgende Reihenfolge: Kuh << Büffel < Ziege < Schaf < Kamel = Stute < Esel < Mensch. Caseinmicellen eignen sich, neben dem natürlichen Transport von Aminosäuren und Mineralstoffen, als Nanotransporter für bioaktive Substanzen wie Vitamin D2 (Semo et al., 2007), β-Carotin (Sáiz-Abajo et al., 2013), Curcumin (Sahu et al., 2008), Triclosan (Roach et al., 2009) oder auch dem antibakteriellen Enzym Lysozym (de Roos et al., 1998; Anema & de Kruif, 2013). Da zu Beginn der Untersuchungen ausschließlich Daten zur Beladung boviner Caseinmicellen mit Hühnereiweißlysozym (HEWL) verfügbar waren, sollte zunächst die Stabilität der tierartspezifischen Micellen und die Effektivität der Beladung mit Lysozym beurteilt werden. Zusätzlich sollte die antibakterielle Wirksamkeit der mit Lysozym beladenen Caseinmicellen verschiedener Säugetiere sowohl in vitro als auch in orientierenden Untersuchungen gegenüber Bakterien erfasst werden. Dabei konnte gezeigt werden, dass die CM aller untersuchten Tierarten mit HEWL beladen werden können. Hinsichtlich der Stabilität der CM sowie der Aufnahmekapazität wurden jedoch tierartspezifische Unterschiede festgestellt. Allgemein konnten die CM aus Wiederkäuer- und aus Humanmilch als stabiler beschrieben werden, wobei die HEWL-Aufnahme bei den CM der Wiederkäuer höher war. Die lytische Enzymaktivität des HEWL in vitro veränderte sich infolge der Micellassoziation für die meisten untersuchten Tierarten im Vergleich zu frei vorliegendem HEWL nicht. Für die antibakterielle Wirksamkeit gegenüber Bacillus subtilis wurden hingegen Unterschiede zwischen den verschiedenen Tierarten beobachtet. Für die HEWL-haltigen CM von Büffel und Ziege ergaben sich die gleichen Wachstumsverzögerungen wie bei freiem HEWL in SMUF. Im Gegensatz dazu wurde die bakteriostatische Wirkung durch die HEWL-Micellen von Kuh, Kamel und Mensch inhibiert, durch die von Schaf, Stute und Esel hingegen verstärkt. Neben der Funktionalisierung der Caseinmicellen durch den Einbau bioaktiver Substanzen, können die Partikel auch chemisch modifiziert werden. Eine Möglichkeit stellt hierbei die Glykierung dar. Glykierungsreaktionen zwischen den Carbonylgruppen reduzierender Zucker und Aminosäureseitenketten wurden für nicht-micellare Caseine in einer Vielzahl an Studien untersucht (Zin El-Din & Aoki, 1993; Morales & van Boekel, 1996; Pellegrino et al., 1999; Lima et al., 2009; Akıllıoğlu & Gökmen, 2014). Über den Einfluss der micellaren Anordnung der Caseine auf die Glykierungsreaktionen war nur wenig bekannt. Erkenntnisse diesbezüglich könnten einen Beitrag zur Aufklärung des Aufbaus der Caseinmicellen leisten. Die Untersuchungen zeigten, dass bei der Erhitzung von micellaren und nicht-micellaren Caseinen für 0 - 4 h bei 100 °C in Gegenwart der reduzierenden Zucker Lactose und Glucose hauptsächlich die frühe Phase der Maillard Reaktion abläuft. Die Bildung der Amadori-Produkte erfolgte dabei in den CM und im nicht micellaren Natrium-Caseinat (NaCas) in gleichem Maße. Hieraus wurde eine ähnliche Zugänglichkeit der reaktiven Aminosäureseitenketten in den micellaren und nicht micellaren Caseinen geschlussfolgert. Signifikante Unterschiede konnten hinsichtlich der Produktbildung der späten Phase der Maillard-Reaktion beobachtet werden. Während Nε-Carboxymethyllysin (CML) im NaCas in höheren Gehalten detektiert wurde, trat in den CM eine verstärkte Pyrralin-Bildung auf. Zudem bewirkte eine Inkubation in Gegenwart von Lactose eine bevorzugte CML-Bildung, wohingegen mit Glucose Pyrralin in höheren Mengen gebildet wurde. Die Maillard-induzierten Proteinquervernetzungsprodukte Glyoxal-Lysin-Dimer und Pentosidin wurden im Vergleich zum zuckerunabhängig gebildeten Lysinoalanin in deutlich geringeren Gehalten erfasst. Hohe Oligomerisierungsgrade zwischen 50 und 84 % zeigten jedoch, dass die Proteinquer-vernetzungen, die im Zuge der Glykierung entstehen, quantitativ von größerer Bedeutung sind. Anhand der Bestimmung der hydrodynamischen Durchmesser und mit Hilfe von REM-Aufnahmen konnte eine weitgehend intra-micellare Proteinquervernetzung abgeleitet werden. Die Ausbildung intra-micellarer Oligomere bewirkte dabei eine höhere Micellstabilität gegenüber einem Calciumentzug durch EGTA. Die vorliegenden Untersuchungen deuten darauf hin, dass die hydrophilen Zuckermoleküle über die von Dalgleish (2011) vorgeschlagenen Wasserkanäle in die CM eindringen und die frühe Phase der Maillard-Reaktion anschließend vor allem in diesen Regionen erfolgt. Die weiteren Phasen der Maillard-Reaktion könnten dann, durch ein tieferes Eindringen der reaktiven Komponenten, zunehmend auch in dehydratisierteren Bereichen ablaufen. Bezüglich des Aufbaus der Caseinmicellen unterstützen die in dieser Arbeit gewonnen Ergebnisse die Vorstellungen des Internal Structure Modells.

The aim of the study was to evaluate the nutritional value and functional properties of several bovine whey proteins concentrate (WPC35 and WPC80), hydrolysate (WPH) and isolate (WPI), casein was used as control. Physicochemical (chemical composition, essential and non-essential aminoacids, minerals) and biological assays were performed to assess the biological quality of proteins and their metabolic responses. After determination of essential and non-essential amino acids, it was found that all protein sources meet the recommendations of FAO/WHO (2007), with the exception of WPC35, probably due to its reduced protein content. The determination of macro and microelements revealed that proteins from bovine milk can be considered not only a source of essential and non-essential amino acids, but also minerals (Ca, Mg and K). Iodine concentration was about one order of magnitude greater for whey proteins and sodium was found in large quantities in these proteins, which can reach 70% and 59% of DRIs, respectively. The biological assay allowed the monitoring of weight gain, food consumption, feed efficiency ratio (FER) and protein intake of animals, evaluate the quality and the metabolic response of protein sources, and the integrity of certain organs and the morphology of the small intestine animals. These studies revealed that all protein sources are of high nutritional value (except WPC35), but larger beneficial biological effects were observed for WPC80, WPH and WPI. Because of the bitter taste of WPH, microencapsulation was made by spray-drying, which significantly improved the taste of panel test (Triangular Test). Considering the presented results, it is concluded that the WPH offers favorable characteristics for use with ingredient in foods for special purposes and that the microencapsulation process yields a significant improvement in flavor, expanding the possibilities for development of foods purposes added hydrolysed proteins, such as infant formula. Some commercial formulations for infants were evaluated with respect to mineral content indicating that the amount of Ca, K and Zn found were considered out of standard of Brazilian law, while the Cu, Mn and Zn levels were very different from those described in label. / O objetivo do estudo foi avaliar o valor nutricional e propriedades funcionais das proteínas do soro de leite bovino concentrado proteico (WPC35 e WPC80), hidrolisado proteico (WPH) e isolado proteico (WPI), além de uma proteína do leite bovino, a caseína, utilizada como controle. Para tanto, foram realizados ensaios físico-químicos (composição centesimal, aminoácidos essenciais e não-essenciais, minerais) e biológicos para avaliar a qualidade proteica e a resposta metabólica das fontes proteicas. Após determinação de aminoácidos essenciais e não-essenciais, foi verificado que todas as fontes proteicas atendem as recomendações estabelecidas pela FAO/WHO (2007), com exceção do WPC35 provavelmente em função do teor reduzido de proteínas. A determinação de macro e microelementos revelaram que as proteínas do leite bovino podem ser consideradas não apenas uma fonte de aminoácidos essenciais e não-essenciais, mas também de minerais (Ca, Mg e K). Para iodo a concentração foi cerca de uma ordem de magnitude superior para as proteínas do soro do leite e sódio foi encontrado em grande quantidade nas proteínas do soro do leite, podendo atingir até 70% e 59% das DRIs, respectivamente. O ensaio biológico permitiu acompanhar o ganho de peso, consumo alimentar, quociente de eficácia alimentar (QEA) e proteína ingerida dos animais, avaliar a qualidade proteica e a resposta metabólica das fontes proteicas, além da integridade de alguns órgãos e histomorfometria do intestino delgado dos animais. Estes estudos revelaram que todas as fontes proteicas são de alto valor nutricional (com exceção do WPC35), mas que maiores efeitos biológicos benéficos foram observados para WPC80, WPH e WPI. Devido ao sabor amargo do WPH, foi feita a microencapsulação por spray-drying, que melhorou de forma significativa o sabor na análise sensorial (Teste Triangular). Considerando-se os resultados apresentados, conclui-se que, o WPH contempla características favoráveis para ser utilizado com ingrediente em alimentos para fins especiais e que o processo de microencapsulação proporcionou uma melhoria significativa no sabor, ampliando as possibilidades de desenvolvimento de alimentos adicionados de proteínas hidrolisadas, tais como fórmulas infantis. Algumas formulações comerciais para lactentes foram avaliadas com relação ao teor de minerais indicando que a quantidade de Ca, K e Zn encontradas foram consideradas fora dos padrões da legislação brasileira, enquanto que os teores de Cu, Mn e Zn determinados foram muito diferentes daqueles descritos no rótulo.

As of today, there is a challenge amongst the elderly to get the energy they need. Malnutrition is a fact for too many and protein is one of the most common deficiencies among macronutrients in this age group. Another group, also in the need of extra energy and protein, are those with a higher level of physical activity. Aventure AB in Lund, Sweden, has earlier developed an “in between meal beverage”, based on oats, with high energy and protein content called “Skaka &amp; Smaka”. This beverage has with promising results been distributed to selected hospitals in Sweden as a pilot-trial. As an expansion of this product, Aventure wanted to create an oat-based, energy dense stirred yogurt. The aim of this study was to investigate the feasibility of addition of different protein sources to create an energy dense stirred yogurt with good texture and a balance of macronutrients. Four different types of protein sources were, separately or in combination, added to the original recipe of “Skaka &amp; Smaka” (without added flavour), after which it was fermented and evaluated regarding sensory and physio-chemical properties. Three different fruit purée mixtures were added separately to the fermented oat-base in different proportions and evaluated by taste, color and flavour intensity. Salt concentration was adjusted and two final products were developed and evaluated through a sensory consumer test, at two different retirement homes in Hässleholm, Sweden. To measure the participants opinions a 9-point hedonic scale was used and attributes evaluated were first impression, color, taste, consistency, thickness, spoonability and total impression. All of the 11 individuals who participated were at the age of 75 and over, where of 2 were men and 9 were women. The proportions of added protein in the two final products were 10% casein and 90% whey. The two selected fruit purées, “skogen” and “havet”, were added in the concentration of 30%. The addition of salt was increased with 100%, resulting in a final concentration of 0.08 g per 100 grams. The sensory analysis revealed no statistical significance between any of the related attributes of the two yogurts. The attribute most important for general liking was taste, followed by texture and color.    Seven individuals (64%) said they could consume this kind of product a few times a week and all participants thought that there was a need for this kind of product. The two yogurts developed in this project, “skogen” and “havet”, and the concept behind them seem to have good potential for consumer liking, though further development of taste and texture is needed. / Som det ser ut idag så finns det en utmaning bland de äldre att få i sig den energi de behöver. Undernäring är ett faktum för allt för många och protein är en av de vanligaste bristerna bland makronäringsämnena inom denna åldersgrupp.    En annan grupp som också är i behov av extra energi och protein är de med en hög fysisk aktivitetsnivå. Aventure AB i Lund, Sverige, har tidigare utvecklat en mellanmåls dryck, baserad på havre, med ett högt energi- och proteininnehåll, kallad ”Skaka &amp; Smaka”. Denna dryck har i ett pilot försök och med lovande resultat distribuerats till flertalet utvalda sjukhus i Sverige. Som en vidareutveckling av denna produkt ville Aventure skapa en havrebaserad, energität, rörd yoghurt. Syftet med denna studie var att undersöka geomförbarheten i tillsättning av olika proteinkällor för att skapa en energität, rörd yoghurt med bra konsistens och en balans av makronäringsämnen. Fyra olika typer av proteinkällor tillsattes, separat eller i kombination, till originalreceptet för ”Skaka &amp; Smaka” (utan tillsatt smak), vartefter blandningen fermenterades och utvärderades utifrån sensoriska och fysio-kemikaliska egenskaper. Tre olika fruktpurée blandningar tillsattes separat till den fermenterade havrebasen i olika proportioner och utvärderades utifrån smak, färg och smakintensitet. Saltkoncentrationen justerades och två slutliga produkter utvecklades och utvärderades genom ett sensoriskt konsumenttest, vid två olika äldreboenden i Hässleholm, Sverige. För att mäta deltagarnas åsikter användes en 9-punkt hedonisk skala och de attribut som utvärderades var första intryck, färg, smak, konsistens, tjockhet, skedbarhet och totalt intryck. Alla de 11 individer som deltog var av ålder 75 och över, varav två var män och nio var kvinnor. Proportionerna av tillsatt protein i de två slutliga produkterna var 10% kasein och 90% vassle. De två valda fruktpuréerna, ”skogen” och ”havet”, tillsattes i en koncentration på 30%. Tillsatsen av salt ökades med 100%, vilket resulterade i en slutlig koncentration på 0.08g per 100 gram. Den sensoriska analysen visade inte på någon statistisk signifikant skillnad mellan de relaterade attributen för de två olika yoghurtarna. Attributen som var av störst vikt för generellt tycke var smak, följt av konsistens och färg. Sju individer (64%) sa att de kunde konsumera en produkt som denna ett par gånger i veckan och alla deltagande tyckte att det fanns ett behov av en produkt som denna. De två yoghurtarna som utvecklades i detta projekt, ”skogen” och ”havet”, och det bakomliggande konceptet tycks ha god potential bland konsumenter, men vidare utveckling av smak och konsistens är nödvändigt.

The study of DNA templated events is not complete without considering the chromatin environment. Histone modifications help to regulate gene expression, chromatin compaction and DNA replication. Because DNA damage repair must occur within the context of chromatin, many remodeling enzymes and histone modifications work in concert to enable access to the DNA and aid in restoration of chromatin after repair is complete. CK2 has recently been identified as a histone modifying enzyme. In this study we identify CK2 as a histone H3 tail kinase in vitro, identify the phospho-acceptor site in vitro, and characterize the modification in vivo in S. cerevisiae. We also characterize the DNA damage phenotype of a strain lacking a single catalytic subunit of CK2. We further characterize the CK2- dependent phosphorylation of serine 1 of histone H4 in vivo. We find that it is recruited directly to the site of a DSB and this recruitment requires the SIN3/RPD3 histone deacetylase complex. We also characterize the contribution of H4 serine 1 phosphorylation in chromatin compaction by using reconstituted nucleosomal arrays to study folding in the analytical ultracentrifuge.

The theme of this diploma thesis is the monitoring of influences on economic aspect of the production of the blue cheese "Hermelín" in the company Pribina. The objective was to specify the dependencies of the deviations of the features of productive cow's milk and of cheese curd made from it and their economics impacts. For this purpose, the analyses of curd grain, whey and cheeses just formed were executed. The samples were taken and analyzed during the manufacturing of cheeses in the factory. Another goal was to try to design a quick operational method for measuring the firmness of curd grain, which is a factor theoretically dependent on renneting qualities of milk and indicating losses of valuable components from cheese curd to whey. The firmness of grain along with these losses may cause negative deviations from required standard of final products, especially at parametres needed for entry of cheeses on sale, such as total solids, weight or content of fat. Concerning productive pasteurize milk, the most attention was focused on the milk coagulation time and its influence on successive phases of manufacturing and on final product. Milk coagulation time is an essential technological quality of milk and it is participant on quantitative and qualitative production of cheese factory. It has been proved that milk coagulation time is notably affected by its titrating acidity, statistic analyse found this influence as very statistically significant (P

The diploma thesis is dealing with design of analytical method for determination of caseins in cow‘s milk. Capillary electrophoresis was used as separative method. The developed procedure was verified on real samples. Theoretical part gives basic information about caseins. Their chemical and physical properties, structure, way of manufacturing, usage in food industry and non-food applications are described. There are further some nutritional factors mentioned, whose application in cattle nutrition can affect milk composition, especially milk protein concentration. Literary part contains summary of capillary electrophoresis methods used for casein determination. Experimental part describes used separative system, procedure of casein isolation from milk and casein content in individual milk samples. Separation of milk caseins was performed in an untreated fused-silica capillary with dimensions of 50 um i.d. x 96 cm total length (71 cm effective length). Phosphate buffer (10 mM, pH 2,5) served as the background electrolyte. Casein from milk was obtained by isoelectric precipitation at pH 4,6 after adding 10% acetic acid. Product was washed with acetone to remove fat and dried. Total casein yield was the highest in bio-milk (27,04 g/l), the lowest in skim long-life milk (22,04 g/l). The diploma thesis has arisen on the basis of cooperation of Institute of Food Science and Biotechnology, Faculty of Chemistry, Brno University of Technology and Research Institute for Cattle Breeding, Ltd., Department of Animal Nutrition Physiology in Pohořelice.