Résumé : Le métabolisme monocarboné ou métabolisme 1-C désigne l’ensemble des voies métaboliques permettant la synthèse et / ou le recyclage de molécules donneur de groupement monocarboné au cours des réactions de méthylation. L’objectif de ce travail était d’évaluer l’implication des métabolismes de la choline, de la phosphatidylcholine (PC) et de la bétaïne dans la physiopathologie des désordres impliquant le métabolisme 1-C en période prénatale et postnatale. Nous avons montré une augmentation progressive de l’expression de la majorité des gènes impliqués dans le métabolisme 1-C au cours de l’ontogénèse hépatique murine, tandis que les leur expression était plus faible avec des profils plus variable au niveau cérébral. Chez l’homme, les valeurs normales des concentrations des intermédiaires du métabolisme 1-C dans le liquide amniotique (LA) en fonction du terme gestationnel ont été déterminées pour tous les paramètres et les concentrations de S-adénosyl-homocystéine et de méthionine étaient augmentées dans les LA du groupe affecté par des défauts de fermeture du tube neural (DFTN) suggérant que certains cas de DFTN pourraient être associés à des déséquilibres du métabolisme 1-C. En post natal, nous avons montré à la fois chez l’homme et l’animal, que les hyperhomocystéinémie d’origine nutritionnelles ou génétiques induisaient une déplétion en bétaïne, épargnant uniquement le rein où elle est un osmolyte majeur. Dans un modèle murin de déficit en cystathionine–beta synthase induisant une hyperhomocystéinémie, une technique de lipidomique ciblée a montré au niveau hépatique des modifications qualitatives des phospholipides (PLs) avec une diminution des PC contenant des acides gras insaturés et des phosphatidyléthanolmines contenant de l’acide arachidonique. Ces modifications des PLs pourraient jouer un rôle dans la constitution de la stéatose hépatique observée dans l’histoire naturelle de cette maladie. En conclusion, ce travail a permis de montrer que la choline, la bétaïne et les PC sont des acteurs indissociables du métabolisme 1-C qui pourraient être impliqués dans la physiopathologie des DFTN et dans les hyperhomocystéinémies. Ils pourraient également être impliqués dans la physiopathologie des stéatoses hépatiques non alcooliques ou des déficits cognitifs, dans lesquels des désordres du métabolisme 1-C ont été observés. / Abstract: One carbon metabolism or 1C metabolism includes all metabolic pathways for the synthesis and / or recycling of molecules involved in methylation reactions. The objective of this study was to evaluate the involvement of choline, phosphatidylcholine (PC) and betaine metabolisms in the pathophysiology of diseases with impaired 1-C metabolism in prenatal and postnatal period. We showed a progressive increase of the expression of the majority of genes involved in 1-C metabolism during the mouse liver ontogeny while their gene expression was at lower levels and with more variable patterns during brain ontogeny. In humans, amniotic fluid concentrations of all intermediates of 1-C metabolism according to gestational term were determined and we observed increased concentrations of S-adenosyl-homocysteine and methionine in pregnancies affected by neural tube defects (NTD) suggesting that some NTDs cases could be associated with an imbalance in 1-C metabolism. In the postnatal period we showed that both in animal and humans and both in nutritional and genetic hyperhomocysteinemia, that betaine pools were decreased, only sparing the kidney betaine concentrations, where betaine acts as an essential osmolyte. In a mouse model of cystathionine-beta synthase deficiency inducing hyperhomocysteinemia, a technic of targeted lipidomic revealed qualitative changes in the liver phospholipids composition, in particular a decrease of PC containing unsaturated fatty acids and of phosphatidylethanolamine containing arachidonic acid and an increase of phosphatidylethanolamine containing docosohaexaenoic acid. This phospholipids remodelage may participate in the development of the steatosis observed in the natural history of this disease. In conclusion, this study has shown that choline, betaine and phosphatidylcholine are essential actors of 1-C metabolism that could be involved in the pathogenesis of NTD and hyperhomocystéinemia. They could also be involved in the pathophgysiology of non alcoholic fatty liver disease or cognitive decline, in which disorders of 1-C metabolism were observed.

The purpose of this research was to quantify properties of strandboard panels manufactured with various preservatives at loading levels effective against native termites. Panels were manufactured using nine different formulations. The method of preservative addition was also examined for some preservative formulations, increasing the total number of preservative treatments to twelve. Panels were manufactured with one target retention for each preservative treatment. An effective preservative loading relative to termites was established by previous studies or referencing current standards. Mechanical testing performed included static bending and internal bond. Physical testing included water absorption, thickness swell, and linear expansion. Few treatments met the Canadian standards for strandboard, but several preservatives performed well. Copper naphthenate, bifenthrin, and copper betaine each deserve further investigation to optimize manufacturing variables to meet required mechanical and physical properties.

Tesis por compendio / En la presente Tesis Doctoral se utilizaron cebada y remolacha como fuentes alternativas de carbohidratos en dietas para trucha arcoíris con el objetivo de evaluar su efecto sobre parámetros productivos, histología hepática e intestinal, estrés y calidad de la carne. Los marcadores estudiados fueron los mismo tanto para los experimentos de cebada como de remolacha. En las pruebas de la cebada se utilizaron concentraciones crecientes de este ingrediente (0-32%) en la dieta, se inició con un peso medio de 127.72 ± 5.65 g y se finalizó cuando alcanzaron el peso comercial al cabo de 84 días. En las pruebas de remolacha se utilizaron dos niveles de remolacha (14% y 28%) y dos de betaína (0.9% y 1.63%), se inició con un peso medio de 69 ± 2.2 g y se finalizó cuando los peces alcanzaron el peso comercial al cabo de 105 días. La inclusión de cebada en dietas de trucha arcoíris no mostró diferencias significativas en los parámetros de crecimiento ni biométricos. La inclusión de 14% remolacha y 0.9% betaína no afectó al crecimiento, parámetros nutritivos, biométricos y retenciones de nutrientes en comparación con el control, concentraciones mayores de remolacha y betaína tuvieron un efecto negativo. Cuando se estudiaron los parámetros histológicos y morfométricos en hígado e intestino, los resultados mostraron que los peces alimentados con concentraciones crecientes de cebada mostraron hepatocitos más pequeños, mientras que los peces alimentados con remolacha y betaína presentaban hepatocitos más grandes, en ambos casos el hígado mostró un menor nivel de vacuolización. La concentración de cebada de un 8% produjo un efecto negativo a nivel morfológico del intestino, sin embargo, la inclusión de remolacha no produjo ningún efecto. Los peces fueron sometidos a estrés por ausencia de oxígeno y aumento de densidad durante 10 minutos. Cuando el estrés fue analizado a nivel bioquímico los resultados mostraron que a niveles basales, la inclusión de cebada y remolacha no causó ningún cambio en los niveles de cortisol, glucosa y MDA, aunque los valores basales de lactato fueron significativamente más bajos en los peces alimentados con remolacha y betaína. Treinta minutos después del estrés la concentración de cortisol, glucosa y lactato aumentó significativamente en los peces de todos los grupos y la concentración de MDA disminuyó. La inclusión de cebada mostró valores más bajos de cortisol, glucosa y lactato que la dieta control. Cuando los peces alcanzaron el peso comercial se analizó la calidad de los filetes y las propiedades antioxidantes. En ambas pruebas los peces alimentados con los ingredientes objeto de estudio mostraron valores de actividad de agua más bajos y una mejora en la textura y el color de los filetes comparado con los peces alimentados con la dieta control. La inclusión de remolacha y betaína no tuvo ningún efecto sobre los parámetros oxidativos del filete, mientras que la inclusión de cebada inhibió su oxidación lipídica. Concentraciones de cebada por encima del 8% mejoraron la actividad antioxidante de los filetes y aumentaron el contenido en alpha-tocoferol, sin embargo, los peces alimentados con remolacha y betaína mostraron un mayor contenido en flavonoides y fenólico pero no se observó ningún efecto sobre las propiedades antioxidantes del filete. Por último, cuando se llevó a cabo el análisis sensorial se observó que los peces alimentados con dietas con más de un 8% de cebada mostraron unas agallas más rojas y mejor textura, además de un color rojo del filete más intenso, parámetros que se correlacionan con una mejora de la frescura del pescado. Sin embargo, la inclusión de cebada o remolacha en dietas de trucha arco iris no tuvo ningún efecto sobre la aceptabilidad de los filetes. / The current Doctoral Thesis used barley and red beet as alternative carbohydrate sources in rainbow trout diets. The aim was to evaluate their effect on productive, histological and morphometric parameters, their effect on biochemical indexes after an acute stress and their effect on the quality of rainbow trout (Oncorhynchus mykiss). The different markers studied were the same in the barley and red beet experiments. In the barley experiment, increasing concentrations of barley (0-32%) were used in the diet, starting with an average weight of 127.72 ± 5.65 g and ending when they reached commercial weight after 84 days. In the red beet experiment, two red beet levels (14% and 28%) and two betaine levels (0.9% and 1.63%) were used, starting with an average weight of 69 ± 2.2 g and finishing when they reached commercial weight at the end of 105 days. The inclusion of barley in rainbow trout diets did not show significant differences in growth and biometric parameters. While the inclusion of 14% of red beet and 0.9% betaine did not affect growth, nutritive or biometric parameters and nutrient retentions compared to control, higher red beet and betaine concentrations showed a negative effect. When the histological and morphometric parameters were studied in liver and intestine, the results showed that fish fed at high barley concentrations showed smaller hepatocytes than control, while hepatocytes were larger in fish fed with red beet and betaine than control, however, in both cases, the liver had a lower level of vacuolization. Barley inclusion at 8% produced a negative effect at intestine morphological level; however, no effects were observed with the inclusion of red beet. Fish were submitted to stress, a lack of oxygen and increased of fish density, during 10 minutes. The results showed that at basal levels, the inclusion of barley and red beet did not cause any change in cortisol, glucose and MDA levels, although basal lactate values were significantly lower in the fish fed with red beet and betaine. Thirty minutes after stress the concentration of cortisol, glucose and lactate increased significantly in the fish of all groups and the concentration of MDA decreased. The inclusion of barley showed lower cortisol, glucose and lactate values than control. When fish reached commercial weight, the quality of the fillets and antioxidant properties were analysed. In both trials, fish fed with the experimental ingredients showed lower water activity values and textural and colour properties were enhanced. Red beet and betaine inclusion did not show any effect on the oxidative parameters of the fillets, while the inclusion of barley showed an inhibitory effect on fillets lipid oxidation. Concentrations of barley above 8% improved the antioxidant activity of the fillets and increased the content of alpha-tocopherol, however, fish fed with red beet and betaine showed a higher content of flavonoids and phenolics but no effect on the antioxidant properties of the fillet. Finally, when the sensory analysis was carried out, it was observed that fish fed diets with more than 8% barley showed redder gills and better texture than control, in addition to a more intense red colour of the fillet, these characteristics are correlated with an improvement of fish freshness. However, the inclusion of barley or red beet in rainbow trout diets had no effect on the acceptability of fillets. / En la present tesi doctoral es van utilitzar ordi i remolatxa com a fonts alternatives de carbohidrats en dietes per a truita amb l'objectiu d'avaluar el seu efecte sobre paràmetres productius, histologia hepàtica i intestinal, estrès i qualitat de la carn. Els marcadors estudiants fòren els mateixos en els experiments d`ordi I remolatxa. En les proves de l'ordi es van utilitzar concentracions creixents d'aquest ingredient (0-32%), es va iniciar amb un pes mitjà de 127.72 ± 5.65 g i es va finalitzar quan van arribar al pes comercial al cap de 84 dies. En les proves de remolatxa es van utilitzar dos nivells de remolatxa (14% i 28%) i dos de betaïna (0.9% i 1.63%), es va iniciar amb un pes mitjà de 69 ± 2.2 g i es va finalitzar quan els peixos van aconseguir el pes comercial al cap de 105 dies. La inclusió d'ordi en dietes de truita no va mostrar diferències significatives en els paràmetres de creixement i biomètrics. La inclusió de 14% remolatxa i 0,9% betaïna no va afectar el creixement, paràmetres nutritius, biomètrics i retencions de nutrients en comparació amb el control, concentracions majors de remolatxa i betaïna van tenir un efecte negatiu. Quan es van estudiar els paràmetres histològics i morfomètrics en fetge i intestí, els resultats van mostrar que els peixos alimentats amb concentracions creixents d'ordi tenien hepatòcits més petits, mentres que els peixos alimentats amb remolatxa i betaïna presentaven hepatòcits més grans, i en ambdós casos el fetge va mostrar un menor nivell de vacuolització. La concentracion d'ordi d'un 8% va produir un efecte negatiu a nivell morfològic de l'intestí, mentres que la inclusió de remolatxa no va produir cap efecte a nivell d'intestí. Els peixos van ser sotmesos a estrès per absència d'oxigen i augment de densitat durant 10 minuts. Quan l'estrès va ser analitzat a nivell bioquímic els resultats van mostrar que a nivells basals, la inclusió d'ordi i remolatxa no va causar cap canvi en els nivells de cortisol, glucosa i MDA, encara que els valors basals de lactat van ser significativament més baixos en els peixos alimentats amb remolatxa i betaïna. 30 minuts després de l'estrès la concentració de cortisol, glucosa i lactat va augmentar significativament en els peixos de tots els grups i la concentració de MDA va disminuir. La inclusió d`ordi va mostrar valors més baixos de cortisol, glucosa i lactat que la dieta control. Quan els peixos van aconseguir el pes comercial es va analitzar la qualitat dels filets i propietats antioxidants. En ambdues proves els peixos alimentats amb els ingredients objecte d'estudi van mostrar valors d'activitat d'aigua més baixos i una millora en la textura i el color dels filets comparat amb els peixos alimentats amb la dieta control. La inclusió de remolatxa i betaïna no va tenir cap efecte sobre els paràmetres oxidatius del filet, mentre que la inclusió d'ordi va inhibir l'oxidació lipídica dels filets. Concentracions d'ordi per sobre del 8% van millorar l'activitat antioxidant dels filets i van augmentar el contingut en alpha-tocoferol, però, els peixos alimentats amb remolatxa i betaïna van mostrar un major contingut en flavonoides i fenòlic però no es va observar cap efecte sobre les propietats antioxidants del filet. Finalment, quan es va dur a terme l'anàlisi sensorial es va observar que els peixos alimentats amb dietes amb més d'un 8% d'ordi van mostrar unes ganyes més vermelles i millor textura, a més d'un color vermell del filet més intens, paràmetres que es correlacionen amb una millora de la frescor del peix. No obstant això, la inclusió d'ordi o remolatxa en dietes de truita no va tenir cap efecte sobre l'acceptabilitat dels filets. / Pinedo Gil, J. (2018). Study of novel carbohydrate sources on rainbow trout (Oncorhynchus mykiss) diets [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/107364 / Compendio

No / Surfactants are commonly used as cleansing agents and yet there are concerns they may also have a role in skin irritation. Presently, the lack of suitable methods for quantitative and qualitative analysis of surfactant deposition on skin has hindered the in-depth investigation of such effects. Here, we report the application of reverse phase liquid chromatography electrospray ionisation mass spectrometry (LC/ESI-MS/MS) assays for two surfactants commonly used in consumer products, namely sodium lauryl ether sulphate (SLES) and laurylamidopropyl betaine (LAPB), to a baseline study aiming to assess deposition levels on human skin. The linearity of the assays was established at 3-20 ng, with coefficient of variation below 5%. Detection limits were 100 pg for LAPB and 1 ng for SLES; quantitation limits were 500 pg for LAPB and 2.5 ng for SLES. The baseline study was conducted using a panel of 40 healthy volunteers. Skin extract samples were taken in triplicate from forearms, using ethanol. SLES was detected on most volunteers, with 75% of them having SLES deposits in the range of 100-600 ng/cm2. LAPB was detected on the skin of all volunteers with 85% of them having deposit levels within the concentration range of 1-100 ng/cm2. These results demonstrate the extent to which commonly used surfactants remain on the skin during the day. The analytical methods reported here can be applied to the investigation of surfactants in relation to general skin condition and the development and optimisation of new consumer wash products. / EPSRC

No / Surfactants are commonly used as cleansing agents and yet there are concerns that they may also have a role in skin irritation. The lack of suitable methods for the quantitative and qualitative analysis of surfactant deposition on skin has hindered the in-depth investigation of such effects. Here, we report the application of reversed-phase liquid chromatography/electrospray ionisation tandem mass spectrometry (LC/ESI-MS/MS) assays for two surfactants commonly used in consumer products, namely sodium lauryl ether sulfate (SLES) and laurylamidopropyl betaine (LAPB), to a baseline study aiming to assess deposition levels on human skin. The linearity of the assays was established at 3-20 ng, with coefficient of variation below 5%. The detection limits were 100 pg for LAPB and 1 ng for SLES; quantitation limits were 500 pg for LAPB and 2.5 ng for SLES. The baseline study was conducted using a panel of 40 healthy volunteers. Skin extract samples were taken in triplicate from forearms, using ethanol. SLES was detected on most volunteers, with 75% of them having SLES deposits in the range of 100-600 ng/cm(2). LAPB was detected on the skin of all volunteers with 85% of them having deposit levels within the concentration range of 1-100 ng/cm(2). These results demonstrate the extent to which commonly used surfactants remain on the skin during the day. The analytical methods reported here can be applied to the investigation of surfactants in relation to general skin condition and to the development and optimisation of new consumer wash products. / EPSRC-DTA award / School Life Sciences

The aim of the present study was the development of analytical methods which can be used for gaining a deeper insight into the mechanisms of hair treatment with cosmetic formulations like shampoos. The efficacy of such a formulation is affected by the amount of different active agents remaining on the hair after treatment. Due to the complex composition of most of the formulations various dependencies and interactions of the ingredients are possible. Beneath that a variety of parameters of the treatment process influence the substantivity. Finally the degree of hair damage present at the treatment process is significant. So the process of hair treatment with cosmetic formulations is very complex. The understanding of this process enables a systematic development of products with better or novel effectivity. The developed analytical methods provide the possibility of a parallel determination of different active agents and their interaction of the adsorption behaviour. Furthermore the determination of influencing variables like treatment conditions or hair damage can be performed. So it is possible to generate the knowledge necessary for excellent scientific product development. The general methodology for the method development consists of an extraction from hair followed by a selective and sensitive quantitation of the active agents located in the extract using LC/MS and LC/MS/MS. Analytical methods for the determination of the complex composed surface active agents Sodium Laureth Sulfate, Cocamidopropyl Betaine and Alkyl Polyglucoside were developed. During the evaluation process it could be figured out that significant swelling of the hair fibre does not occur during a hair wash lasting 2 min or less. Hence the active agents are mainly adsorbed at the hair surface. During method development it was found that the composition of these agents is changing during hair treatment resulting in a difference of the singles substance distribution on hair and in the shampoo. Additionally an analytical method for selective and sensitive quantitation of the cationic polysaccharides Polyquaternium-10 and Guar Hydroxypropyltrimonium Chloride using LC/MS was developed. The parallel quantitation of Polyquaternium-10 and Guar Hydroxypropyltrimonium Chloride enables to study their interaction of the adsorption behaviour. Beneath that the quantitation of substances with low active contentes, like Ubiquinone 50, Piroctone Olamine and Benzophenone?4, was carried out. The extraction behaviour of these substances showed that an extraction time of 7 min assisted by an ultrasonic treatment is optimal. Longer extraction leads to a significant increase of the active hair surface due to swelling. The result is a higher adsorption capacity of the hair fibre and thus a decreased analyte concentration in the extract. The reproducibility of the using treatment protocol was approved by parallel quantitation of several active agents. Due to variation of several treatment parameters their influence on substantivity was studied: the flow rate, temperature and hardness of the water for rinsing and the time for massaging in the formulation. Furthermore, it was shown that the degree of hair damage and further shampoo ingredients influence the substantivity of active agents. Finally it was demonstrated that the adsorption behaviour can be significantly influenced by the application strategy. / Das Ziel der vorliegenden Arbeit war es, mit der Entwicklung von analytischen Methoden eine Möglichkeit zu schaffen, um die Mechanismen bei der Haarbehandlung mit kosmetischen Formulierungen, wie Shampoos besser verstehen zu können. Die Pflegeleistung einer solchen Formulierung wird unter anderem davon bestimmt, wie viel der unterschiedlichen eingesetzten Wirkstoffe nach der Behandlung auf dem Haar verbleiben. Durch die Komplexität der kosmetischen Formulierungen kommt es zu vielfältigen Abhängigkeiten und Beeinflussungen der Substantivität der einzelnen Substanzen voneinander. Darüber hinaus gibt es eine Vielzahl von Parametern bei der Haarbehandlung, die die Substantivität beeinflussen. Schließlich spielt auch der Zustand des Haares eine entscheidende Rolle. Alles in allem handelt es sich bei der Behandlung des Haares mit kosmetischen Formulierungen um sehr komplexe Prozesse. Das Verständnis dieser Prozesse ermöglicht eine gezielte Entwicklung von Produkten mit besserem und zum Teil neuartigem Leistungsspektrum. Die entwickelten Analysenmethoden erlauben die parallele Bestimmung der auf dem Haar adsorbierten Menge verschiedener Wirkstoffe. Dadurch kann die gegenseitige Beeinflussung der Wirkstoffadsorption untersucht werden. Darüber hinaus ermöglicht das Methodenspektrum eine Bestimmung der Einflussgröße äußerer Parameter, wie der Waschbedingungen und innerer Parameter, wie der Haarschädigung. Damit ist es möglich, das notwendige Wissen für eine fortschrittliche, wissenschaftliche Produktentwicklung zu gewinnen. Der allgemeine Ansatz für die Entwicklung der Methoden beinhaltet eine Extraktion der Wirkstoffe vom Haar, gefolgt von einer selektiven und empfindlichen Quantifizierung im Extrakt mittels LC-MS und LC-MS/MS. Es wurden Analysenmethoden für die komplex zusammengesetzten oberflächenaktiven Wirkstoffe Sodium Laureth Sulfate, Cocamidopropyl Betaine und Alkyl Polyglucoside entwickelt. Dabei wurde durch Untersuchungen der Extraktion vom Haar bewiesen, dass die zweiminütige Haarwäsche nicht zu einem signifikanten Aufquellen des Haares führt, so dass die Wirkstoffe hauptsächlich an der Haaroberfläche adsorbiert werden. Außerdem wurde gezeigt, dass sich die Zusammensetzung dieser Wirkstoffe während der Haarwäsche ändert, so dass sich die Einzelsubstanzverteilung auf dem Haar von der im Shampoo unterscheidet. Des Weiteren wurde die selektive und empfindliche Quantifizierung der kationischen Polymere Polyquaternium-10 und Guar Hydroxypropyltrimonium Chloride durchgeführt. Die parallele Quantifizierung beider Konditionierungsstoffe ermöglicht die Untersuchung, in welchem Umfang sich beide Polymere in ihrem Adsorptionsverhalten beeinflussen. Bei der Methodenentwicklung für die Quantifizierung der Wirkstoffe Ubiquinone, Piroctone Olamine und Benzophenone-4 zeigte sich, dass eine Extraktion über 7 min im Ultraschallbad die besten Ergebnisse brachte. Bei längerer Extraktionszeit vergrößert sich die Oberfläche des Haares durch Quellung, so dass sich der Anteil an adsorbierten Analyten wieder erhöht. Die Reproduzierbarkeit der Haarwäsche mit dem verwendeten Waschprotokoll wurde bestätigt, indem verschiedene Wirkstoffe parallel auf dem Haar quantifiziert wurden. Durch Veränderung der Waschparameter Einmassierzeit sowie Durchflussmenge, Temperatur und Härte des Spülwassers wurde der Einfluss dieser Parameter auf die Substantivität der Wirkstoffe untersucht. Darüber wurde gezeigt, dass sowohl der Schädigungsgrad des Haares als auch andere auf dem Haar befindliche Shampoo-Bestandteile einen Einfluss auf die Substantivität von Wirkstoffen haben. Schließlich wurde an einem Beispiel gezeigt, dass man die Substantivität von Wirkstoffen nicht nur durch die Zusammensetzung der kosmetischen Formulierung, sondern auch durch die Art der Anwendung entscheidend beeinflussen kann.

The aim of the present study was the development of analytical methods which can be used for gaining a deeper insight into the mechanisms of hair treatment with cosmetic formulations like shampoos. The efficacy of such a formulation is affected by the amount of different active agents remaining on the hair after treatment. Due to the complex composition of most of the formulations various dependencies and interactions of the ingredients are possible. Beneath that a variety of parameters of the treatment process influence the substantivity. Finally the degree of hair damage present at the treatment process is significant. So the process of hair treatment with cosmetic formulations is very complex. The understanding of this process enables a systematic development of products with better or novel effectivity. The developed analytical methods provide the possibility of a parallel determination of different active agents and their interaction of the adsorption behaviour. Furthermore the determination of influencing variables like treatment conditions or hair damage can be performed. So it is possible to generate the knowledge necessary for excellent scientific product development. The general methodology for the method development consists of an extraction from hair followed by a selective and sensitive quantitation of the active agents located in the extract using LC/MS and LC/MS/MS. Analytical methods for the determination of the complex composed surface active agents Sodium Laureth Sulfate, Cocamidopropyl Betaine and Alkyl Polyglucoside were developed. During the evaluation process it could be figured out that significant swelling of the hair fibre does not occur during a hair wash lasting 2 min or less. Hence the active agents are mainly adsorbed at the hair surface. During method development it was found that the composition of these agents is changing during hair treatment resulting in a difference of the singles substance distribution on hair and in the shampoo. Additionally an analytical method for selective and sensitive quantitation of the cationic polysaccharides Polyquaternium-10 and Guar Hydroxypropyltrimonium Chloride using LC/MS was developed. The parallel quantitation of Polyquaternium-10 and Guar Hydroxypropyltrimonium Chloride enables to study their interaction of the adsorption behaviour. Beneath that the quantitation of substances with low active contentes, like Ubiquinone 50, Piroctone Olamine and Benzophenone?4, was carried out. The extraction behaviour of these substances showed that an extraction time of 7 min assisted by an ultrasonic treatment is optimal. Longer extraction leads to a significant increase of the active hair surface due to swelling. The result is a higher adsorption capacity of the hair fibre and thus a decreased analyte concentration in the extract. The reproducibility of the using treatment protocol was approved by parallel quantitation of several active agents. Due to variation of several treatment parameters their influence on substantivity was studied: the flow rate, temperature and hardness of the water for rinsing and the time for massaging in the formulation. Furthermore, it was shown that the degree of hair damage and further shampoo ingredients influence the substantivity of active agents. Finally it was demonstrated that the adsorption behaviour can be significantly influenced by the application strategy. / Das Ziel der vorliegenden Arbeit war es, mit der Entwicklung von analytischen Methoden eine Möglichkeit zu schaffen, um die Mechanismen bei der Haarbehandlung mit kosmetischen Formulierungen, wie Shampoos besser verstehen zu können. Die Pflegeleistung einer solchen Formulierung wird unter anderem davon bestimmt, wie viel der unterschiedlichen eingesetzten Wirkstoffe nach der Behandlung auf dem Haar verbleiben. Durch die Komplexität der kosmetischen Formulierungen kommt es zu vielfältigen Abhängigkeiten und Beeinflussungen der Substantivität der einzelnen Substanzen voneinander. Darüber hinaus gibt es eine Vielzahl von Parametern bei der Haarbehandlung, die die Substantivität beeinflussen. Schließlich spielt auch der Zustand des Haares eine entscheidende Rolle. Alles in allem handelt es sich bei der Behandlung des Haares mit kosmetischen Formulierungen um sehr komplexe Prozesse. Das Verständnis dieser Prozesse ermöglicht eine gezielte Entwicklung von Produkten mit besserem und zum Teil neuartigem Leistungsspektrum. Die entwickelten Analysenmethoden erlauben die parallele Bestimmung der auf dem Haar adsorbierten Menge verschiedener Wirkstoffe. Dadurch kann die gegenseitige Beeinflussung der Wirkstoffadsorption untersucht werden. Darüber hinaus ermöglicht das Methodenspektrum eine Bestimmung der Einflussgröße äußerer Parameter, wie der Waschbedingungen und innerer Parameter, wie der Haarschädigung. Damit ist es möglich, das notwendige Wissen für eine fortschrittliche, wissenschaftliche Produktentwicklung zu gewinnen. Der allgemeine Ansatz für die Entwicklung der Methoden beinhaltet eine Extraktion der Wirkstoffe vom Haar, gefolgt von einer selektiven und empfindlichen Quantifizierung im Extrakt mittels LC-MS und LC-MS/MS. Es wurden Analysenmethoden für die komplex zusammengesetzten oberflächenaktiven Wirkstoffe Sodium Laureth Sulfate, Cocamidopropyl Betaine und Alkyl Polyglucoside entwickelt. Dabei wurde durch Untersuchungen der Extraktion vom Haar bewiesen, dass die zweiminütige Haarwäsche nicht zu einem signifikanten Aufquellen des Haares führt, so dass die Wirkstoffe hauptsächlich an der Haaroberfläche adsorbiert werden. Außerdem wurde gezeigt, dass sich die Zusammensetzung dieser Wirkstoffe während der Haarwäsche ändert, so dass sich die Einzelsubstanzverteilung auf dem Haar von der im Shampoo unterscheidet. Des Weiteren wurde die selektive und empfindliche Quantifizierung der kationischen Polymere Polyquaternium-10 und Guar Hydroxypropyltrimonium Chloride durchgeführt. Die parallele Quantifizierung beider Konditionierungsstoffe ermöglicht die Untersuchung, in welchem Umfang sich beide Polymere in ihrem Adsorptionsverhalten beeinflussen. Bei der Methodenentwicklung für die Quantifizierung der Wirkstoffe Ubiquinone, Piroctone Olamine und Benzophenone-4 zeigte sich, dass eine Extraktion über 7 min im Ultraschallbad die besten Ergebnisse brachte. Bei längerer Extraktionszeit vergrößert sich die Oberfläche des Haares durch Quellung, so dass sich der Anteil an adsorbierten Analyten wieder erhöht. Die Reproduzierbarkeit der Haarwäsche mit dem verwendeten Waschprotokoll wurde bestätigt, indem verschiedene Wirkstoffe parallel auf dem Haar quantifiziert wurden. Durch Veränderung der Waschparameter Einmassierzeit sowie Durchflussmenge, Temperatur und Härte des Spülwassers wurde der Einfluss dieser Parameter auf die Substantivität der Wirkstoffe untersucht. Darüber wurde gezeigt, dass sowohl der Schädigungsgrad des Haares als auch andere auf dem Haar befindliche Shampoo-Bestandteile einen Einfluss auf die Substantivität von Wirkstoffen haben. Schließlich wurde an einem Beispiel gezeigt, dass man die Substantivität von Wirkstoffen nicht nur durch die Zusammensetzung der kosmetischen Formulierung, sondern auch durch die Art der Anwendung entscheidend beeinflussen kann.

<p>Cilj ove doktorske disertacije predstavljalo je ispitivanje uticaja vodnog stresa izazvanog&nbsp; sa polietilen glikolom (PEG)&nbsp; 6000 na biohemijske osobine klonova topole. Reznice tri klona topole (M-1, B-229 i PE19/66) su hidroponski gajene i izložene 100 mOsm i 200 mOsm PEG 6000&nbsp; tokom &scaron;est dana. Nakon tretmana u kontrolnim i stresiranim<br />biljkama su&nbsp; ispitane promene aktivnosti antioksidantnih enzima,&nbsp; aktivnost dva<br />enzimska markera polifenolnog metabolizma, fenolni profil, antioksidantna aktivnost, kao i sadržaj prolina i glicin betaina&nbsp; (GB). Takođe, ispitan je biohemijski odgovor kulture tkiva klona M-1 na vodni stres izazvan sa PEG 6000.</p><p>U oba eksperimenta uočeno je da je akumulacija prolina i GB u uslovima vodnog stresa izazvanog sa PEG 6000&nbsp;najvažnija strategija u otpornosti na stres i prevazilaženju vodnog deficita, tako da se&nbsp;ovi parametri mogu definisati kao najbolji indikatori otpornosti topole na su&scaron;u.</p><p>Rezultati ovog istraživanjaupotpunjavaju saznanja o povezanosti oksidativnog stresa uzrokovanog vodnim stresom sa antioksidantnim odgovorom, nivoom o&scaron;tećenja lipida i proteina, uključivanjem osmolita kao i promenama u polifenolnom profilu. Takođe, dobijeni rezultati su značajni za odabir klonova topola tolerantnih na vodni stres&nbsp; i&nbsp; za<br />po&scaron;umljavanje su&scaron;nih stani&scaron;ta, kojih će biti sve vi&scaron;e usled negativnog uticaja klimatskih promena.</p> / <p>The aim of presented doctoral thesis was investigation of the impact of water deficit caused by polyethylene glycol (PEG) 6000 on biochemical features of poplar clones.&nbsp; During six days, cuttings of three poplar clones (M-1, B-229 and PE 19/66) were grown in hydroponics and subjected to 100 and 200 mOsm PEG 6000. Changes in the activity of antioxidant enzymes, polyphenol characterization, two enzymatic markers of polyphenol metabolism, antioxidant capacity, as well as proline and glycine betaine content were investigated in stressed plants. Furthermore,&nbsp; assessment of&nbsp; biochemical response of poplar clone M-1 tissue culture to water stress induced by PEG 6000 was performed.</p><p>In both experiments, the most important strategy for stress resistance and overcoming water deficiency was accumulation of proline and glycine betaine so these parameters can be defined as the best indicators of poplar resistance to drought.</p><p>The results of this study complete&nbsp; the findings on the oxidative stress caused by water stress&nbsp; and their relationship with the antioxidant response, the level of lipid andprotein damage, accumulation of the osmolites and changes in the polyphenol profile. Also, the obtained results may be important for the selection of poplar clones resistant to water stress and for afforestation in arid sites, which appeared to be more numerous due to the effects of&nbsp; global climate change.</p>

Bakgrund: Syftet med denna studie var att undersöka effekten av sju dagars tillskott av betain (2,5 gram/dag) på prestationsförmågan i idrottsspecifika tester för manliga vältränade cykelatleter. Metod: Sex (n=6) manliga testpersoner (genomsnitt ± standardavvikelse ålder, 31,7 ± 10,2 år; längd, 188,2 ± 3,6 cm; kroppsvikt; 82,5 ± 7,6 kg; fettfri massa, 71,6 ± 7,3 kg) genomförde förtester bestående av fem counter movement jumps, fyra stycken 12 s wingate-sprinter med 2,5 minuters aktiv vila följt av fem ytterligare counter movement jumps. De blev sedan indelade slumpmässigt i betain- eller placebogrupper. Första testet etablerade basvärden och eftertest ”1” samt ”2” föregicks med antingen sju dagars tillskott med placebo (2,5 g maltodextrin plus 12,5 mg riboflavin) eller betain (2,5 g trimetylglycin plus 12,5 mg riboflavin). Riboflavin användes för att undanröja den fiskliknande doften som kan uppstå vid tillskott av betain. Doseringen bestod av fyra kapslar per dag till måltid, varav två kapslar intogs på morgonen eller förmiddagen (före träning) och två kapslar intogs på eftermiddagen eller kvällen (efter träning). Vi använde oss av en dubbelblindad, kontrollerad randomisering cross-over design; med en veckas washout mellan placebo och betain. Maximum Peak Power, Average Peak Power, Maximum Average Power, Mean Average Power, Mean Relative Power Drop, Maximum Relative Power Drop, CMJ-Medel och CMJ-Max före och efter sprinterna analyserades med two-tailed t-test 2 och 3 för att jämföra skillnader före och efter intag av tillskott samt skillnaderna mellan grupperna.Resultat: Inga signifikanta skillnader kunde noteras. Konklusion: En veckas tillskott av betain gav inte någon signifikant förbättring jämfört med placebo i hopp- och sprintförmåga samt förmåga att ta ut sig själv maximalt hos aktiva cyklister. Fler studier med framförallt större testgrupper efterlyses för att utvärdera betainets ergogena effekt bland aktiva atleter. / The effect of betaine supplementation on counter movement jumps and sprint performance among male bicycle athletes Background: The purpose of this study was to investigate the effect of a seven days supplementation regimen with betaine (2,5 grams per day) on counter movement jumps and sprint performance in trained male bicyclists. Method: Six (n=6) male subjects (mean ± standard deviation age, 31,7 ± 10,2 years; height, 188,2 ± 3,6 cm; bodyweight, 82,5 ± 7,6 kg; lean body mass, 71,6 ± 7,6 kg) completed a 21 day study involving five counter movement jumps (no arm swing) followed by 4 consecutive modified 12 second Wingate sprints on a bicycle ergometer with active rest (2,5 minutes) followed by another set of five counter movement jumps. A double blind, controlled randomized cross over design was used and there was a 1 week washout between the supplementation startup for each group. Following pre-testing (baseline) the participants were randomly divided into groups by choosing one of the following supplements: “12” Placebo –2,5 grams of maltodextrine plus 12,5 mg riboflavin or “11” – Active 2,5 grams of trimethylglycine plus 12,5 mg riboflavin. Riboflavin was used so mask the “fishy odor syndrome” effect of high intakes of trimethylglycine that may cause trimethyluria. Daily consumption of capsules was divided into two doses of two capsules each with meals. One dose (two capsules) was consumed in the morning and one dose in the evening. The following variables were analyzed: Maximum Peak Power, Average Peak Power, Maximum Mean Power, Average Mean Power, Maximum Relative Power Drop, Average Relative Power Drop, Mean CMJ-Pre + Post sprints and Maximal CMJ-Pre + Post sprints. Results: None of the parameters showed statistical significance. Conclusion: Our study indicates that seven days of betaine ingestion does not seem significantly improve counter movement jump or sprint capacity in male trained bicyclists. Larger studies are needed to investigate the performance enhancing effects of betaine, especially among high performance athletes.

Submitted by Claudinei Pracidelli (cpracide@ipen.br) on 2016-11-11T09:28:13Z No. of bitstreams: 0 / Made available in DSpace on 2016-11-11T09:28:13Z (GMT). No. of bitstreams: 0 / O monocloroacetato de sódio (MCAS) e o dicloroacetato de sódio (DCAS) são compostos tóxicos e irritantes ao ser humano e nocivos ao meio ambiente, sendo impurezas indesejáveis na cocoamido propil betaína (CAPB), que é um surfactante anfótero utilizado em produtos de consumo dos segmentos cosmético e domiciliar. Diante dos requisitos de concentração em nível de mg/kg exigidos pelos órgãos reguladores de saúde do governo, tornou-se mandatório o emprego de metodologia com limite de quantificação, precisão e exatidão adequados aos rígidos controles de processo pelos fabricantes da CAPB, bem como, dispor de técnicas convencionais com poder de resolução e proficiência pelo controle de qualidade e neste contexto inseriu-se a cromatografia a gás. Neste estudo foram estabelecidos os procedimentos analíticos que definiram as melhores condições para identificar e quantificar as impurezas MCAS e DCAS na matriz CAPB por meio da cromatografia a gás. A preparação das amostras consistiu da derivação das impurezas MCAS e DCAS a ésteres etílicos e a extração líquido-líquido em hexano para separar dos demais constituintes da matriz. Os modos de detecção acoplados à cromatografia a gás foram a ionização pela chama (GC/FID), a captura de elétrons (GC/ECD) e a espectrometria de massas (GC/MS). A validação comprovou que as metodologias são lineares entre 4 e 50 mg/kg com recuperação de 70 a 120%, apresentam limites de quantificação inferiores a 10 mg/kg e produziram médias e incertezas similares na amostra examinada, constituindo-se alternativas para a determinação de cloroacetatos em betaínas. / Dissertação (Mestrado em Tecnologia Nuclear) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP