Introdução: O Estudo Longitudinal de Saúde do Adulto é um estudo de coorte multicêntrico com o objetivo de identificar os fatores de risco associados ao diabetes tipo 2 e à doença cardiovascular na população brasileira. Nosso principal objetivo é explicar a concepção e implementação das rotinas do laboratório central e biobanco do Estudo Longitudinal de Saúde do Adulto (ELSA-Brasil); destacando as forças e limitações do protocolo. O segundo objetivo é descrever os pré-testes usados para validar o protocolo de antes do início do estudo. Métodos: O estudo optou pela centralização dos exames em um laboratório central no Hospital Universitário da Universidade de São Paulo (USP), Com base em dados recentes que confirma a estabilidade da glicose em amostras congeladas, até os testes de glicose no sangue foram centralizadas. Porém o processamento das amostras foi realizado nos laboratórios locais o que reduziu os custos do transporte das amostras para o laboratório central. A estratégia implicou na necessidade de implementação de equipes nos laboratórios em cada Centro de Investigação para coletar e processar as amostras antes do transporte. O estudo incluiu exames para a avaliação do metabolismo da glicose, resistência à insulina, perfil lipídico, eletrólitos, ácido úrico, hormônios da tireóide, função hepática e contagem total de células do sangue. Além disso, as amostras de DNA, urina plasma, e de soro foram colhidas e Além desses exames, o estudo também extraiu DNA de leucócitos, colheu e estocou amostras de urina, plasma e soro. Para garantir a homogeneidade do protocolo todos os membros das equipes locais passaram por treinamento e certificação centralizado, com visitas cruzadas entre os centros durante o campo para controle de qualidade. Resultados: A opção por um laboratório central garantiu a uniformidade da metodologia utilizada para a realização dos exames, evitando as variações inevitáveis entre laboratórios. Durante 26 meses, cerca de 375.000 testes foram realizados no laboratório central. Não houve perda de amostras biológicas durante o estudo. A implementação do biobanco usando palhetas armazenados em repositórios de nitrogênio foi realizada sem problemas importantes desde 2008 até agora. Conclusão: O ELSA-Brasil mostrou a exequibilidade de exames multicêntricos no Brasil com todos os exames realizados em um laboratório central, de uma maneira custo-efetiva. A logística de armazenamento de amostras biológicas foi feito com custos aceitáveis e de qualidade, sendo um modelo para estudos futuros / Background: The Longitudinal Study of Adult Health is a multicenter cohort study aimed to identify risk factors associated with type 2 diabetes and cardiovascular disease in Brazilian population. Our main objective is to explain the conception and implementation of the routines of the central laboratory and biobank of the Longitudinal Study of Adult Health (ELSA-Brasil) highlighting the strength and the limitations of the protocol The second objective is to describe the pre-tests used to validate the protocol before the start of the study. Methods: The study made an option to centralize the exams in one central laboratory at the University Hospital, São Paulo University (\"USP\"). Based on recent data that confirms the stability of glucose in freeze samples, even blood glucose tests were centralized. However, biological samples were processed in the local laboratories, reducing the weight of the material to be transported, and diminishing costs of transportation to the central lab. Especially trained local teams collected and processed biological samples before transportation. The study included tests for glucose metabolism, insulin resistance, lipid profile, electrolytes, uric acid, thyroid hormones, hepatic function, and total blood cell count. In addition, DNA, urine, plasma and serum samples were collected and stored. In order to guarantee protocol homogeneity, all team members underwent centralized training and certification, and cross-visits in each research center were done. Results: The choice of a central laboratory assured uniformity of the methodology used for the exams, avoiding the variations between laboratories During 26 months, approximately 375,000 tests were done in the central- laboratory. There was no loss of biological samples during the study. The implementation of the biobank using straws stored in nitrogen repositories was performed without important problems since 2008 until now. Conclusion: The ELSA-Brazil showed the feasibility of a multicenter study in Brazil with all the analyses performed in a central laboratory, in a cost-effective way. The logistic of storage of biological samples was done with acceptable costs and quality being a model for future studies

O desequilíbrio da homeostase emocional tem sido considerado como um mecanismo subjacente aos transtornos de ansiedade e humor. Em fêmeas, as alterações na secreção hormonal durante as diferentes fases do ciclo estral podem ser a base das alterações na reatividade emocional a eventos estressantes. Estudos comportamentais sobre diferenças sexuais no processamento das emoções mostram resultados conflitantes em fêmeas devido às dificuldades na seleção dos melhores modelos animais para testar as diferenças associadas ao ciclo estral. Uma vez que os testes comportamentais foram desenvolvidos em animais do sexo masculino, eles podem não ser apropriados para fêmeas. O presente estudo foi desenvolvido para contribuir nessa linha de pesquisa usando diferentes modelos de animais de medo incondicionado e condicionado, considerando as diferentes fases do ciclo estral das ratas. Comparou-se o desempenho de machos e fêmeas nas quatro fases do ciclo estral em dois testes de medo incondicionado: o switch-off, em que ratos cruzam uma caixa vai-e-vem para desligar uma luz aversiva, e o registro de vocalizações ultrassônicas (VUSs) a 22 kHz emitidos por animais sob o estresse agudo de restrição. Nos testes de medo condicionado, registrou-se o sobressalto potencializado pelo medo e a resposta decongelamento a um contexto aversivo. Em ambos os testes de medo condicionado, a reatividade emocional não se mostrou diferente entre os sexos. No entanto, no que diz respeito ao medo incondicionado, ratas em diestro tardio apresentaram maior reatividade emocional em desligar a luz intensa e maior emissão de VUSs em resposta à restrição em relação a outras fases do ciclo. Estes achados sugerem que o perfil hormonal durante a fase do diestro 2 pode aumentar a reatividade emocional de ratas frente a estímulos inatos, porém não àqueles aprendidos. / Dysfunctional emotional regulation has been implicated as a potential mechanism underlying anxiety and mood disorders. Changes in hormonal secretion during the different phases of the estrous cycle may underlie changes in emotional reactivity to stressful events in female animals. Previous behavioral studies of sex differences in emotion processing in females have yielded conflicting results. This may be due to the range of different behavioral tests used and difficulties in selecting the best animal models to test for estrous cycle-linked differences in responsiveness. Furthermore, the commonly used behavioral tests were developed in male animals and it may not be appropriate to translate directly the protocols from males to females. In the present study we have attempted to address these problems by using different animal models of anxiety based on tests for unconditioned or conditioned fear. We compared the performance of male rats and female rats at four stages of the estrous cycle defined by differences in vaginal cytology. To test for unconditioned fear, we used two tests: a light switch- off test, in which rats escape to the other compartment of a shuttle-box to turn off an aversive light and recordings of 22 kHz ultrasound vocalizations (USVs) during acute restraint stress. For the conditioned fear paradigm, we used fear potentiated startle in an aversive context and conditioned freezing using an aversive context as the conditioned stimulus. In both tests of conditioned fear there were no gender or estrous cycle-linked differences in emotional reactivity. However, with respect to unconditioned fear, female rats in late diestrus showed greater emotional reactivity expressed as switch-off responses to a light environment and USVs in response to restraint compared to other phases of the cycle. These findings suggest that the hormonal profile during the late diestrous phase may predispose to up-regulated emotional reactivity in rats facing emotional challenges to unconditioned, but not conditioned fear- inducing stimuli.

L'utilisation massive des nanomatériaux pose de réels enjeux sanitaires et environnementaux. C'est pourquoi ils sont désormais soumis à une réglementation qui prévoit une traçabilité de ceux-ci depuis leur fabrication jusqu'à leur distribution et l'établissement d'une fiche d'identité de la substance (composition, taille, état d'agglomération, forme, etc.). Une routine de caractérisation de nanoparticules en suspension a ainsi été développée. La Microscopie Électronique en Transmission (MET) a permis d'établir une majorité des paramètres de la fiche d'identité, en combinant à la fois imagerie et spectroscopie (analyses chimiques). La préparation, dont dépendra la qualité des observations, nécessite un développement pour chaque matériau analysé. Pour cela, trois techniques ont été mises au point : le dépôt en voie sèche qui permet une observation directe et simple, la cryogénie qui permet de fixer l'état de la suspension et l'in-situ liquide qui permet d'observer directement la suspension sans changement d'état. Les analyses MET étant locales, une comparaison avec des techniques indirectes a été effectuée par Diffusion Statique (MALS) et Dynamique (DLS) de la Lumière avec et sans fractionnement par couplage flux-force (FFF). Deux matériaux modèles ont été choisis. Le premier est une nanoémulsion de lipides stabilisés par des surfactants, servant de vecteurs à des principes actifs. Une étude de vieillissement par interaction avec des protéines a été menée et de légères variations de taille ont été obtenues. Le second matériau sélectionné est une poudre de nanoparticules de dioxyde de titane, remises en suspension, utilisée dans les crèmes solaires en tant que filtres UV. Ces particules ont été observées avant et après passage en enceinte climatique afin d'observer les effets des rayons UV sur celles-ci. Ceci a confirmé la stabilité des particules. Les protocoles de caractérisation développés au cours de cette thèse peuvent ainsi servir de supports à l'étude d'autres nanoparticules en suspension. / The extensive use of nanomaterials has raised awareness about health issues and their fate in the environment. That is why they are now subject to regulation that has imposed their traceability from their manufacturing to their distribution as the establishment of their characteristics (chemical composition, size, agglomeration state, shape ...). A characterization routine for nanoparticles in suspension was developed. Transmission Electron Microscopy (TEM) fulfills most of the criteria cited before by combining imaging and spectroscopy techniques. Three sample preparation methods were optimized to ensure high quality results : a dry process, rapid freezing to vitrify the sample and the use of an textit{in-situ} liquid TEM holder to prevent any preparation artefact (no phase change). To obtain quantitative analysis, a comparison was made between Dynamic Light Scattering (DLS), Multi-Angle Light Scattering (MALS), with and without a fractionation system (AF4), and TEM. To support this work, two nanomaterials were analyzed. The first one is a nanoemulsion composed of lipid nanoparticles stabilized by surfactants used as nanocarriers for drug delivery. Their stability after protein interaction was investigate and some size variations were observed. The second material is a powder composed of titanium dioxide nanoparticles used as UV filters in sunscreens. These nanoparticles were analyzed before and after interaction with UV radiation in a climatic chamber to confirm their stability. The different protocols developed in this PhD may be used for the analysis of other nanomaterials.

Made available in DSpace on 2015-04-22T22:17:28Z (GMT). No. of bitstreams: 1 Dissertacao - Eyner Godinho de Andrade.pdf: 1620387 bytes, checksum: d94449ab292c6a6a393caf5983969e46 (MD5) Previous issue date: 2006-08-03 / Fundação de Amparo à Pesquisa do Estado do Amazonas / The aim of this study was to evaluate the effect of the frozen storage on the sensory, chemical, microbiological stability and functional properties of the protein of minced fish of tambaqui (Colossoma macropomum) and matrinxã (Brycon amazonicus) catering by fish culture located near Manaus (AM), elaborated in laboratories conditions. In general, during frozen storage of minced fish occurs losses on a functionality of proteins as reduction of solubility and of water-holding capacity (WHC), as well as deterioration enzymatic and bacterial, when diverse volatile composites that can incite, among others problems are formed, the oxidative rancidity and the muscular increase of pH. To evaluate the stability of minced of matrinxã and tambaqui during its storage -20ºC had been used parameters as: values of N-BVT, pH, sensory tests and bacteriological countings. The protein solubility and the WHC had been determined to verify the possible alterations in the functionality of proteins due to the freezing. Also the chemical and nutritional characterization of minced was made fish by means of the proximal composition and amino acid profile. The income of the triturated meat was determined, after gotten the processes of decapitation, gutted and separation mechanics. The results had been low: 37.44% and 47.0% for tambaqui and matrinxã, respectively. On the basis of the text of lipids and energy value, both the fish had been classified as semi fat. Tambaqui with 2,66% of lipids and 90,98 kcal and matrinxã with 5,55% of lipids and energy of 127,67% kcal. The fish used in the experiment process high proteins text: 16.74% for tambaqui and 18.61% for matrinxã, being gifts in its composition all the essential amino acids. The protein solubility of minced fish for both the studied species presented a drastic reduction in the first month of storage and it tends to stabilize sixty days after, remaining itself it enters 0,03 and 0,04 g/dL until the end of the experiment. Throughout the one hundred and fifteen days of frozen stockage -20ºC minced fish of tambaqui and matrinxã had presented stability of the CRA that varied between 95 and 97%. The N-BVT, for both minced , presented an increase in first the thirty days, followed of a fall accented until the end of the experiment, being always below of the limit of 30 mg of N-BVT/100 g. The values of pH had shown to a rise of 6,17 the 6,39 for matrinxã and of 6,30 the 6,48 for tambaqui until the sixty days of storage -20ºC and after this period had a fall with trend to the stability. During the frozen storage of minced of matrinxã a reduction of the bacterial countings was noticed. It can have had contamination of the samples or imperfections in the freezing, not having, however, the presence of Escherichia coli. In the sensory evaluation the products of both the species had been remained in quality A throughout the experimental period. The results allow to conclude that minced fish had kept same stability for long periods of frozen storage consisting in an alternative for diversification of the production of the regional fishing industry and allowing the search for other markets, national as in such a way international. / O objetivo deste trabalho foi avaliar os efeitos da estocagem congelada sobre a estabilidade sensorial, química, microbiológica e propriedades funcionais da proteína dos minced fish de tambaqui (Colossoma macropomum) e de matrinxã (Brycon amazonicus) provenientes de pisciculturas localizadas próximas à Manaus (AM), elaborados em condições laboratoriais. De modo geral, durante a estocagem congelada dos minced fish ocorrem perdas na funcionalidade das proteínas como a diminuição da solubilidade protéica e da capacidade de retenção de água (CRA), bem como a deterioração enzimática e bacteriana, quando são formados diversos compostos voláteis que podem provocar, entre outros problemas, a rancidez oxidativa e o aumento do pH muscular. Para avaliar a estabilidade dos minced de matrinxã e tambaqui durante sua estocagem a -20ºC foram utilizados parâmetros como: valores de N-BVT, pH, testes sensoriais e contagens bacteriológicas. A solubilidade protéica e o CRA foram determinados para verificar as possíveis alterações na funcionalidade das proteínas devido ao congelamento. Foi também feita a caracterização química e nutricional dos minced fish por meio da composição centesimal e perfil de aminoácidos. Foi determinado o rendimento da carne triturada, obtida após os processos de decaptação, evisceração e separação mecânica. Os resultados foram baixos: 37,44% e 47,0% para o tambaqui e a matrinxã, respectivamente. Com base no teor de lipídios e valor energético, ambos os peixes foram classificados como semigordos. O tambaqui com 2,66% de lipídios e 90,98 kcal/100g e a matrinxã com 5,55% de lipídios e energia de 127,67 kcal/100g. Os peixes utilizados no experimento possuíam alto teor protéico: 16,74% para o tambaqui e 18,61% para a matrinxã, estando presentes em sua composição todos os aminoácidos essenciais. A solubilidade protéica dos minced fish para ambas as espécies estudadas apresentou uma diminuição drástica no primeiro mês de armazenagem e tendeu a estabilizar-se após sessenta dias, mantendo-se entre 0,03 e 0,04 g/dL até o final do experimento. Ao longo dos cento e quinze dias de estocagem congelada a -20ºC os minced fish de tambaqui e matrinxã apresentaram estabilidade do CRA que variou entre 95 e 97%. O N-BVT, para ambos os minced , apresentou um aumento nos primeiros trinta dias, seguido de uma queda acentuada até o final do experimento, ficando sempre abaixo do limite de 30 mg de N-BVT/100 g. Os valores de pH mostraram uma elevação de 6,17 a 6,39 para a matrinxã e de 6,30 a 6,48 para o tambaqui até os sessenta dias de estocagem a -20ºC e após esse período houve uma queda com tendência à estabilidade. Durante a estocagem congelada do minced de matrinxã notou-se uma redução das contagens bacterianas. Pode ter havido contaminação das amostras ou falhas no congelamento, não havendo, contudo, a presença de Escherichia coli. Na avaliação sensorial os produtos de ambas as espécies mantiveram-se em qualidade A ao longo do período experimental. Os resultados permitem concluir que os minced fish mantiveram estabilidade mesmo por longos períodos de estocagem sob congelamento constituindo-se em uma alternativa para diversificação da produção da indústria pesqueira regional e permitindo a busca por outros mercados, tanto nacional como internacional.

Durante situações aversivas, como choque nas patas, imobilidade ou restrição dos movimentos, há um aumento nos níveis de acetilcolina no hipocampo de ratos. Além disso, o aumento desse neurotransmissor está envolvido com a modulação comportamental do MCC, principalmente em sua porção dorsal, HD. Diante disso, é proposto neste estudo que os receptores muscarínicos presentes no HD de ratos, modulam a expressão da REC, no modelo do MCC. Ratos wistar foram submetidos às sessões de condicionamento aversivo contextual, sendo divididos nos grupos: condicionado (6 choques de 3s; 1,5 mA) e não condicionado (sem choques) . Quarenta e oito horas após esta sessão foram avaliadas a comportamental (congelamento) e respostas autonômicas (PAM, FC e queda da TC). Além disso, foi verificada a distribuição de receptores M1 e M3 nas subáreas do HD e a quantificação dos mesmos em animais condicionados, não condicionados e naive, 48 após a sessão de condicionamento. A microinjeção bilateral de hemicolínio, inibidor da captação de colina, no HD promoveu uma redução do tempo de congelamento durante a reexposição ao contexto aversivo, caracterizando um efeito do tipo ansiolítico. Além disso, este tratamento inibiu o aumento da PAM, FC e a queda da TC induzidos pelo MCC. O mesmo ocorreu com a administração de atropina, antagonista não seletivo de receptores muscarínicos, de forma dose-dependente em todas as respostas observadas. Adicionalmente, a microinjeção de diferentes doses de J104129 fumarate bloquearam o tempo de congelamento (de forma dose-dependente, semelhante a atropina), a elevação da PAM, FC e a queda da TC durante a re-exposição ao contexto aversivo. Como o J104129 fumarate bloqueia receptores muscarínicos tanto M1 quanto M3, foi utilizado um antagonista de maior afinidade para receptores do tipo M1, pirenzepina. Todas as doses utilizadas de pirenzepina inibiram as respostas autonômicas, sem afetar o tempo de congelamento induzidos pelo MCC. A análise de imunofluorescência, por duplamarcação, mostrou que receptores M1 e M3 estão distribuídos nos mesmos compartimentos celulares nas subáreas do HD. A quantificação dos receptores 48 horas após o condicionamento evidencia apenas o aumento de receptores M3 no hipocampo de ratos, sem alteração na população de receptores M1. Com este conjunto de resultados podemos concluir que a ACh no HD é essencial para a expressão da REC. Especificamente, eles sugerem que os receptores muscarínicos do tipo M1 presentes nesta estrutura estão envolvidos com as respostas autonômicas e somente os receptores M3 participam das respostas comportamentais. / During aversive and stressful situations, such as footshock, stillness and restriction of movements, there is an increase in the levels of acetylcholine in rat hippocampus. Moreover, the increasing in the synaptic level of this neurotransmitter is involved with behavioral modulation of contextual fear conditioning (CFC), especially in the dorsal portion, DH. Therefore, this study investigated the involvement of muscarinic receptors in DH of rats in the expression of conditioned emotional response (CER) in the CFC. Moreover, we verified the expression of these receptors in the DH by double labeling immunofluorescence. Male Wistar rats were subjected to aversive contextual conditioning sessions and were divided into two groups: conditioned and unconditioned. Forty-eight hours after this session were evaluated autonomic (mean arterial pressure, MAP, heart rate, HR and tail temperature, TT) and behavioral (freezing) responses. Moreover, the distribution of M1/M3 receptors in subareas of DH was observed and the quantification of these receptors were performed 48 hours after the conditioning session in conditioned, unconditioned and naive animals. The bilateral microinjection of hemicholinium, inhibitor of choline reuptake, in DH, caused a decrease of freezing during re-exposure to the aversive context, featuring an anxiolytic-like effect. Furthermore, this treatment inhibited the increase in MAP, HR and TT drop by CFC. DH muscarinic receptors antagonism evoked by atropine, a non-selective muscarinic antagonist, reduced the freezing, in a dose-dependent manner. Similar reduction was observed in autonomic responses. The selective antagonism of M1/M3 receptors evoked by J104129 fumarate also reduced freezing, in a dose-dependent manner, when compared with control animals. The same effect was observed with autonomic responses in all the tested doses. As J104129 fumarate blocks both M1 muscarinic as M3 muscarinic, was used a higher-affinity M1 antagonist, pirenzepine. All doses inhibited cardiovascular responses and decrease the TT drop, without affecting the freezing induced by CFC. The immunofluorescence analysis revealed that M1 and M3 receptors are distributed in the same cellular compartments in DH. The quantification of receptors showed an increase of M3 receptors in rat hippocampus, while no change in the density of receptors M1 was detected. These findings support that cholinergic neurotransmission present in DH is involved with the expression of responses evoked by fear contextual conditioning, through muscarinic receptors activation. In particular, M3 muscarinic receptors modulate behavioral responses, M1 and M3 muscarinic receptors modulate the autonomic responses.

La maladie de Parkinson induit des troubles locomoteurs complexes et variés. Des troubles spécifiques de la marche tels que le " freezing " peuvent survenir ponctuellement. Afin d'améliorer l'accompagnement de ces patients, il est nécessaire d'améliorer notre compréhension de ces situations. Cette thèse vise à caractériser l'activité locomotrice des patients parkinsoniens en situation réelle. Un travail interdisciplinaire a été mené, fondé sur une approche écologique. Quatorze patients ont été suivis. La méthode déployée intégrait le recueil de marqueurs biomécaniques et physiologiques, de l'expérience vécue ainsi que des observations directes. La description de leur expérience a révélé une perception sensori-motrice altérée et une organisation du comportement locomoteur spécifique à l'environnement. Les analyses montrent un schéma d'activation musculaire spécifique, avec notamment une prolongation générale de l'activité des muscles pendant la phase d'appui simple et une augmentation de cocontraction lorsque le pied est en contact au sol. Les résultats de cette analyse intégrée du comportement moteur ont permis de caractériser une typologie de cinq marches et de trois types de " freezing ". De plus, ces observations ont mis en valeur des stratégies utilisé par des patients ainsi qu'un mécanisme d'ajustement neuromusculaire pour augmenter la stabilité de la marche. A long terme, ce travail devrait favoriser 1) la compréhension des bases neurologiques des troubles de la marche ; 2) la prise en charge des patients parkinsoniens ; 3) l'identification des biomarqueurs de la stabilité de la marche ; 4) la spécification des systèmes embarqués du suivi de la marche. / Patients with Parkinson’s disease (PD) experience complex locomotor disturbances. Their walking abilities can be highly variable and specific problems such as freezing of gait may arise during daily life activity. A greater understanding of such difficulties is required in order to develop new therapeutic strategies. This thesis examines the emergence of PD gait patterns during everyday situations. To this end, this study used an interdisciplinary approach drawing upon ecological and embodied perspectives of human activity. The investigation focused upon fourteen PD patients. Data collection methods combined interviews, direct observations and onboard measures of physiological and biomechanical performance. This research identified important changes in patient sensorimotor perception during gait and specific principles regarding the organisation of their movements with respect to the environment. Analysis revealed characteristic changes to lower limb muscle activation patterns, most notably the prolongation of muscle activity during the mid-stance phase; and an increase in cocontraction of agonist-antagonist muscles across the ankle and knee whilst the leg was in contact with the ground. This integrated approach enabled the characterisation of five walking patterns and three types of freezing. These results highlighted patient self-management strategies, and indicated the role of neuromuscular mechanisms for increasing gait stability. Ultimately, these findings may assist to 1) understand the neurological bases of gait dysfunction; 2) enhance PD patient care; 3) identify biomarkers for gait stability and; 4) support the development of ambulatory monitoring technologies.

<p>Superchilling is a method that can be used to prolong the shelf life of foods by partial freezing. Knowledge of the effects of this method on both the shelf life and quality of foods is important in order to find optimal processing and storage conditions and is of great importance both for the industry and for the consumers. Different raw materials of muscle foods were studied with the purpose of creating a basis for further improvements of both the process and the storage conditions. Products from the commercially important species pork (both roasts and rib steaks), Atlantic salmon (both vacuum packed and wrapped fillets) and Atlantic cod (vacuum packed fillets) were chosen to represent different muscle foods.</p><p>Based on both sensory and microbial evaluations, the superchilled storage improved shelf life of pork roasts from 2 to 16 weeks, and shelf life might even be further prolonged if temperature is kept more constant. The H2S-producing bacteria in superchilled cod fillets did not exceed the limit of consumption during the whole storage period of six weeks, while the microbial shelf life of the ice chilled fillets was estimated to be 8 days after processing.</p><p>Quality changes have been studied with focus on biochemical and physiochemical properties. One of the major goals in meat and fish processing is the ability to retain water and it was observed that drip loss was lower in superchilled samples compared to traditionally chilled samples in all species studied. However, the subsequent liquid loss was higher both in pork roasts and in cod fillets.</p><p>The extractability of protein was used as a tool to monitor protein denaturation. It was found that myofibrillar proteins denatured more easily during superchilled than during traditionally chilled storage both in salmon and in cod fillets. This was confirmed by electrophoresis in salmon. The amount of free amino acids were higher in cod than in salmon and increased more during superchilled storage than during ice chilled storage indicating exoproteolytic activity during storage. Activities of cathepsins B and B + L in salmon fillets were relatively stable during the storage period in all storage groups, demonstrating that these enzymes are not deactivated at the selected storage temperatures and may therefore lead to softening during subsequent chilled storage.</p><p>In superchilled samples of all species studied, white spots emerged on the product surface. However, the appearance of spots did not correspond either to higher drip loss or to higher microbial growth on surface of these samples. On the contrary, the total plate counts of superchilled samples were lower than of the other storage groups. This observation also applied to iron agar counts on cod fillets. These findings are interpreted as a strong indication of that the spots most likely were not of microbial origin.</p><p>The optimal superchilling process and storage conditions remains to be found for the products studied. From the present results it is reasonable to suggest that optimal storage temperature for the vacuum packed salmon fillets is found between the superchilled temperatures in the present experiments. The quality both of the pork roasts and the cod fillets would probably benefit from a storage temperature slightly closer to 0 °C than those studied. It can also be assumed that the control of the superchilling process is more essential to cod than to salmon and pork. In addition, the properties of the raw material are crucial in order to obtain high quality of products after processing and storage.</p> / Paper I-III are reprinted with kind permission from Elsevier, sciencedirect.com

Superchilling is a method that can be used to prolong the shelf life of foods by partial freezing. Knowledge of the effects of this method on both the shelf life and quality of foods is important in order to find optimal processing and storage conditions and is of great importance both for the industry and for the consumers. Different raw materials of muscle foods were studied with the purpose of creating a basis for further improvements of both the process and the storage conditions. Products from the commercially important species pork (both roasts and rib steaks), Atlantic salmon (both vacuum packed and wrapped fillets) and Atlantic cod (vacuum packed fillets) were chosen to represent different muscle foods. Based on both sensory and microbial evaluations, the superchilled storage improved shelf life of pork roasts from 2 to 16 weeks, and shelf life might even be further prolonged if temperature is kept more constant. The H2S-producing bacteria in superchilled cod fillets did not exceed the limit of consumption during the whole storage period of six weeks, while the microbial shelf life of the ice chilled fillets was estimated to be 8 days after processing. Quality changes have been studied with focus on biochemical and physiochemical properties. One of the major goals in meat and fish processing is the ability to retain water and it was observed that drip loss was lower in superchilled samples compared to traditionally chilled samples in all species studied. However, the subsequent liquid loss was higher both in pork roasts and in cod fillets. The extractability of protein was used as a tool to monitor protein denaturation. It was found that myofibrillar proteins denatured more easily during superchilled than during traditionally chilled storage both in salmon and in cod fillets. This was confirmed by electrophoresis in salmon. The amount of free amino acids were higher in cod than in salmon and increased more during superchilled storage than during ice chilled storage indicating exoproteolytic activity during storage. Activities of cathepsins B and B + L in salmon fillets were relatively stable during the storage period in all storage groups, demonstrating that these enzymes are not deactivated at the selected storage temperatures and may therefore lead to softening during subsequent chilled storage. In superchilled samples of all species studied, white spots emerged on the product surface. However, the appearance of spots did not correspond either to higher drip loss or to higher microbial growth on surface of these samples. On the contrary, the total plate counts of superchilled samples were lower than of the other storage groups. This observation also applied to iron agar counts on cod fillets. These findings are interpreted as a strong indication of that the spots most likely were not of microbial origin. The optimal superchilling process and storage conditions remains to be found for the products studied. From the present results it is reasonable to suggest that optimal storage temperature for the vacuum packed salmon fillets is found between the superchilled temperatures in the present experiments. The quality both of the pork roasts and the cod fillets would probably benefit from a storage temperature slightly closer to 0 °C than those studied. It can also be assumed that the control of the superchilling process is more essential to cod than to salmon and pork. In addition, the properties of the raw material are crucial in order to obtain high quality of products after processing and storage. / Paper I-III are reprinted with kind permission from Elsevier, sciencedirect.com

Plants have spread to almost everywhere in the world. As they disperse, they meet many different environments to which they may be able to adapt. For a plant species to adapt to a new environment, genetic variation is needed. The individuals differ from each other in their genetic composition, which often means differences in phenotypes. Those individuals that manage to reproduce will form the next generation. With different conditions in different environments, it will not be the same phenotypes that reproduce everywhere. In that way, plant species will form into a mosaic of locally adapted populations varying genetically as the species disperses. After the last ice age plants have started to disperse away from the equators. With increasing latitudes come increasing challenges to migrating plants. As plant species disperse northwards along this gradient of varying conditions individuals are selected for cold adaptive traits like flowering time and freezing tolerance, acquired by cold acclimation. In this way, genetic variation from the original populations for these traits becomes sorted out along a latitudinal cline. The aim of this thesis was to understand how selection along a latitudinal gradient has shaped natural variation in cold adaptive traits in plants dispersing northwards, and specifically, to investigate what variation can be observed in phenotypes for these traits and how these traits correlate with genetic variation in genes known to be involved in cold acclimation. In this study significant variation was found in a sample of the model plan Arabidopsis thaliana accessions in cold adaptive traits flowering time and freezing tolerance. A clear latitudinal cline in the cold adaptive traits freezing tolerance for A. thaliana was observed. Analysis of nucleotide polymorphism for the cold responsive ICE1 (inducer of CBF expression 1) transcription factor revealed a haplotype structure with two allelic clades as well as unusually high levels of synonymous polymorphism. Nucleotide polymorphism analysis for the transcription factors CBF1, CBF2 and CBF3 (C-repeat binding factors) that play a key role in regulating the expression of a group of target genes known as the “CBF regulon” showed a distinct geographical haplotype structure. One haplotype was dominant in southern accessions while in the other northern accessions overrepresented. There was a significant effect of CBF haplotype on both freezing tolerance and flowering time even after correcting for latitude. Significant differences in CBF expression levels were found between the different CBF genes as well as between different accessions. Sequence variation at CBF was shown to have a significant effect on expression levels of CBF2. No clear correlations were found between CBF gene expression and freezing tolerance or temperature sensitivity for any of the accessions used in the study. This highlights the complex relationship between sequence variation in candidate genes and gene expression, and the problems associated with unraveling the genetic basis of ecologically important traits.

Invasive non-indigenous species are among the principle drivers of global change, altering nutrient cycles, changing disturbance regimes, and generally threatening biodiversity. Climate change is widely expected to exacerbate invasions by relaxing abiotic barriers, such as low temperature, but the mechanistic evidence supporting this is limited. Here, I evaluate the hypothesis that low temperatures determine the northern range limit of kudzu (Pueraria montana var. lobata), an invasive Asiatic vine in North America, by assessing freezing and chilling tolerance of kudzu plants in winter, spring, summer, and fall. Kudzu was widely planted throughout the southeastern U.S. in the early 20th Century to prevent erosion. It is winter-deciduous and reproduces primarily from buds on stem nodes. In the last 40 years, kudzu has migrated northward in concert with a northward shift in the -20oC minimum winter temperature isocline, indicating that less severe winter cold is permitting northward migration. Freezing mortality during winter does not explain this correlation. Electrolyte leakage assays demonstrate that above- and belowground kudzu stems can survive to -27oC and -17oC. Insulation provided by soil and snow protects belowground stems from lethal temperatures to well north of kudzu's current range limit. Severe spring chill stops growth and photosynthesis and causes some shoot mortality, but both growth and photosynthesis recover quickly following the chill. Summer growth rates are rapid (up to 22 cm/d), responding within hours to temperature changes, and are unimpaired by nighttime lows. Photosynthesis is reduced at cool temperatures, but on cool days, kudzu leaves tend to be warmer than air temperature, so photosynthesis rates generally remain close to optimal values. In autumn, growth stops below 15oC, but leaves are retained and maintain modest photosynthetic competence until killed by frost in November. In colder climates that occur far north of kudzu's current range, reductions in the length and quality of the growing season could accumulate over time to reduce kudzu's success. There is, however, no strong evidence that low temperatures in any season will prevent kudzu from migrating throughout southern Ontario, making kudzu a good candidate for invasive species regulations.