The available literature indicates that to date, few research has been performed on oil powders for topical delivery. The aim of this project was to investigate the release characteristics of oil powder formulations, as well as their dermal and transdermal delivery properties. Whey protein-stabilised emulsions were used to develop oil powders. Whey protein was used alone, or in combination with chitosan or carrageenan. Nine oil powders, with salicylic acid as the active ingredient, were formulated by using the layer-by-layer method. Three different pH values (pH 4, 5 and 6) were used to prepare the formulations, because of the different charges that polymeric emulsifiers may have. The characteristics of the prepared oil powders were determined, including their droplet sizes, particle size distributions, loss on drying, encapsulation efficiencies, oil leakage and water dispersibility. Release studies (membrane diffusion studies) were conducted by utilising cellulose acetate membranes (0.2 μm pore size) and Franz-type diffusion cells over a period of eight hours. The release of the active ingredient was determined for all nine powders, their respective template emulsions, as well as their respective oil powders redispersed in water. The release of salicylic acid from the respective redispersed oil powders was then further compared to its release from the template emulsions and from the oil powders. The effect of pH and different polymer types used in preparing the oil powders, their respective redispersed oil powders and the template emulsions were determined with regards to the release of the active ingredient from all these preparations. The effect of pH and different polymers used was furthermore determined on the oil powders and their respective redispersed oil powders, with regards to their dermal and transdermal deliveries. Transdermal delivery and skin uptake were investigated on specifically selected powders only, based on the outcomes of the oil powder characterisation and release data. The qualifying formulations were chitosan pH 4, 5 and 6, whey and carrageenan pH 6 oil powders, together with their respective redispersed oil powders in water. Human abdominal skin was dermatomed (thickness 400 μm) for use in the diffusion studies. Franz-type diffusion cells were used over a period of 24 hours. The results of the membrane release studies indicated that the oil powders had achieved a significantly higher release than their respective redispersed oil powders. The release of salicylic acid from the redispersed oil powders and from their respective emulsions was similar. The transdermal delivery test outcomes showed that the effect of pH could have been influenced by the degree of ionisation, resulting in a decrease in permeation with increasing ionisation of salicylic acid, in accordance with the pH partition hypothesis. Furthermore, biopolymers, such as chitosan had demonstrated a penetration enhancing effect, which had led to the enhanced dermal and transdermal delivery of salicylic acid. A correlation was also found between the powder particle size and transdermal delivery. / MSc (Pharmaceutics), North-West University, Potchefstroom Campus, 2014

The available literature indicates that to date, few research has been performed on oil powders for topical delivery. The aim of this project was to investigate the release characteristics of oil powder formulations, as well as their dermal and transdermal delivery properties. Whey protein-stabilised emulsions were used to develop oil powders. Whey protein was used alone, or in combination with chitosan or carrageenan. Nine oil powders, with salicylic acid as the active ingredient, were formulated by using the layer-by-layer method. Three different pH values (pH 4, 5 and 6) were used to prepare the formulations, because of the different charges that polymeric emulsifiers may have. The characteristics of the prepared oil powders were determined, including their droplet sizes, particle size distributions, loss on drying, encapsulation efficiencies, oil leakage and water dispersibility. Release studies (membrane diffusion studies) were conducted by utilising cellulose acetate membranes (0.2 μm pore size) and Franz-type diffusion cells over a period of eight hours. The release of the active ingredient was determined for all nine powders, their respective template emulsions, as well as their respective oil powders redispersed in water. The release of salicylic acid from the respective redispersed oil powders was then further compared to its release from the template emulsions and from the oil powders. The effect of pH and different polymer types used in preparing the oil powders, their respective redispersed oil powders and the template emulsions were determined with regards to the release of the active ingredient from all these preparations. The effect of pH and different polymers used was furthermore determined on the oil powders and their respective redispersed oil powders, with regards to their dermal and transdermal deliveries. Transdermal delivery and skin uptake were investigated on specifically selected powders only, based on the outcomes of the oil powder characterisation and release data. The qualifying formulations were chitosan pH 4, 5 and 6, whey and carrageenan pH 6 oil powders, together with their respective redispersed oil powders in water. Human abdominal skin was dermatomed (thickness 400 μm) for use in the diffusion studies. Franz-type diffusion cells were used over a period of 24 hours. The results of the membrane release studies indicated that the oil powders had achieved a significantly higher release than their respective redispersed oil powders. The release of salicylic acid from the redispersed oil powders and from their respective emulsions was similar. The transdermal delivery test outcomes showed that the effect of pH could have been influenced by the degree of ionisation, resulting in a decrease in permeation with increasing ionisation of salicylic acid, in accordance with the pH partition hypothesis. Furthermore, biopolymers, such as chitosan had demonstrated a penetration enhancing effect, which had led to the enhanced dermal and transdermal delivery of salicylic acid. A correlation was also found between the powder particle size and transdermal delivery. / MSc (Pharmaceutics), North-West University, Potchefstroom Campus, 2014

A estrutura e o comportamento de proteínas são de grande interesse a uma infinidade de áreas de conhecimento, com especial destaque para alimentação e saúde. As proteínas compõem uma fundamental classe de moléculas biológicas, com capacidade para interagir com diversas outras estruturas e formar complexos. Dois tópicos principais são abordados neste trabalho: (a) avaliação da formação de complexos entre três proteínas do soro do leite (-lactoalbumina, -lactoglobulina e lactoferrina) em diferentes condições experimentais de força iônica e pH da solução através de derivadas da energia livre e (b) avaliação de aspectos relacionados com a fundamentação teórica dos modelos moleculares com granularidade grossa e uso do dielétrico contínuo empregados para estudo do fenômeno da complexação molecular, através da análise, em larga escala, da correlação de cargas em proteínas disponíveis no banco de dados de proteínas. Os estudos teóricos de complexação foram realizados através do emprego de simulações Monte Carlo Metropolis, com o modelo do solvente contínuo, constante dielétrica homogênea e sal implicitamente descrito. Os estudos de complexação permitem compreender os mecanismos moleculares envolvidos, elucidando, inclusive, controvérsias experimentais e oferecendo orientação para aplicações. Já os resultados do estudo de correlação de cargas em estruturas experimentais de proteínas permitem justificar as aproximações do modelo. / The structure and behavior of proteins are of great interest for a number of science fields, with special emphasis to food industry and health. Proteins comprise a fundamental group of biological molecules, with the capacity of interacting with several other structures and creating complexes. Two major topics are covered in this study: (a) evaluation of complex formation among three whey proteins (-lactalbumin, -lactoglobulin and lactoferrin) under different experimental conditions of ionic strength and pH of solution through free energy derivatives and (b) evaluation of features related to the theoretical basis of the coarse-grained molecular models and the use of continuous dielectric which are employed to investigate the molecular complexation phenomenon, through analysis, on a large scale, of charge correlation in proteins available in the protein data bank. The theoretical studies of complexation were performed through the use of Monte Carlo Metropolis simulations, with continuous solvent model, homogeneous dielectric constant and salt implicitly described. The complexation studies allow to understand the molecular mechanisms involved, also elucidating experimental controversies and offering orientation to applications; whereas the results of the charge correlation study in experimental structure of proteins allow justify the approximations of the model.

Orientador: Flavia Maria Netto / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-09-27T19:42:42Z (GMT). No. of bitstreams: 1 HozUrrejola_LuciadelCarmendela_D.pdf: 2021405 bytes, checksum: 7522596dbd78ae21dcc1d21542fc59d5 (MD5) Previous issue date: 2006 / Resumo: Amostras de ß-lactoglobulina (ß-LG) no estado sólido, com diferentes atividades de água (aw) (0,22; 0,53; 0,74), ou em solução, em concentração de proteína de 3 e 10 mg/mL, foram irradiadas (1-50 kGy) utilizando uma fonte de Cobalto-60. Espalhamento de raios-X a baixos ângulos (SAXS), dicroísmo circular (CD) e fluorescência foram as técnicas utilizadas para estudar as mudanças conformacionais da ß-LG por efeito da irradiação. A proteína irradiada foi também examinada por eletroforese em gel de poliacrilamida (PAGE) e desnaturante (SDS-PAGE) sob condições redutoras e não redutoras, cromatografia de exclusão molecular (SE-HPLC) e ¿SH livre. A ß-LG no estado sólido nas diferentes aw estudadas não foi afetada pela irradiação. As amostras não apresentaram diminuição na digestibilidade sob ação da pepsina e tripsina e não mudaram seu poder ligante de retinol quando irradiadas com doses até 10 kGy. Nas amostras irradiadas em solução, houve modificações estruturais, ao nível de estrutura terciária, enquanto a estrutura secundária parece ter sido preservada, como indicado pela quantificação das estruturas obtidas a partir da deconvolução dos espectros de CD no UV-longínquo. Os dados de SE-HPLC e espalhamento de raios-X mostraram que a irradiação promoveu a formação de oligômeros e agregados, que resultaram ser altamente solúveis, como mostrado pelas medidas de turbidez. Da análise dos dados de SAXS e a construção de modelos pode ser mostrado que os oligômeros eram múltiplos de dímeros de ß-LG, (tetrâmeros e hexâmeros, etc.), linearmente ordenados. Ligações cruzadas intermoleculares entre radicais tirosil teriam participação na agregação e as ligações dissulfeto, pelo contrário, não teriam sido relevantes para a formação dos agregados. O mecanismo de agregação da ß-LG mediante ligações cruzadas de bitirosil, contrário ao que acontece em outros tratamentos físicos, como o térmico ou de alta pressão hidrostática, poderia estar gerando agregados com características e propriedades diferentes. O efeito da irradiação na proteína foi maior nas amostras mais diluídas e com maior dose de radiação, indicação da ocorrência do chamado ¿efeito indireto¿ da irradiação. Considerando que não houve alterações estruturais, nem perda de funcionalidade ou de digestibilidade da ß-LG irradiada no estado sólido até com 10 kGy em diferentes aw, o tratamento com radiação gama poderia ser usado para melhorar a qualidade microbiológica da proteína e aumentar as condições de segurança se for aplicada uma radiação na faixa geralmente utilizada no processamento de alimentos (<10 kGy) / Abstract: ß-lactoglobulin (ß-LG) in solution at different protein concentrations (3 and 10 mg/mL) and in solid state, with different water activity (aw) (0.22; 0.53; 0.74), was irradiated using a Cobalt-60 radiation source at a dose level of 1-50 kGy. Small-angle X-ray scattering (SAXS), Circular Dichroism (CD) and Fluorescence spectroscopy were used to study the conformational changes of ß-LG due the irradiation treatment. The irradiated protein was also examined by polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate (SDS) PAGE under reducing and nonreducing conditions, size exclusion chromatography (SE-HPLC), and free SH. ß-LG in the solid state, at different aw, was not significantly affected by gamma radiation. The digestibility of these samples under pepsin or trypsin action was not diminished, and the retinol binding property was not changed by irradiation up to 10 kGy. The irradiation of ß-LG in solution promoted structural changes at the tertiary structure level but the secondary structure seemed to be maintained, as indicated by the quantification of structures from the deconvolution of the far-UV CD spectra. The SEHPLC, and X-rays scattering data showed that irradiation of ß-LG solutions promoted oligomers and aggregates formation, which proved to be highly soluble, as shown by turbidity measurements. From the SAXS data analyses and the model building it could be shown that the formed oligomers were linear molecules, built by linear combinations of ß- LG dimers (tetramers, hexamers, etc.). Formation of intermolecular cross-linking between tyrosil radicals are proposed to be at least partially responsible by this occurrence, and the disulphide bonds, in contrast, do not appear to be relevant to the formation of these aggregates. The mechanism of aggregation of ß-LG through bityrosil cross-links, differently from the mechanism occurring in other physical treatments as heating or high pressure, could form aggregates with special and different characteristics. The radiation effect was more significant at low protein concentration and higher irradiation dose, being consistent with the occurrence of the ¿indirect effect¿ when proteins are irradiated in solution. Considering that neither structural changes or loss of functionality and digestibility of ß-LG irradiated in the solid state at different aw, with dose up to 10 kGy, was detected, the gamma radiation treatment could be used for improving the microbiological quality of the protein, enhancing the safety conditions when applying the radiation range generally used in food processing (<10kGy) / Doutorado / Doutor em Engenharia de Alimentos

A estrutura e o comportamento de proteínas são de grande interesse a uma infinidade de áreas de conhecimento, com especial destaque para alimentação e saúde. As proteínas compõem uma fundamental classe de moléculas biológicas, com capacidade para interagir com diversas outras estruturas e formar complexos. Dois tópicos principais são abordados neste trabalho: (a) avaliação da formação de complexos entre três proteínas do soro do leite (-lactoalbumina, -lactoglobulina e lactoferrina) em diferentes condições experimentais de força iônica e pH da solução através de derivadas da energia livre e (b) avaliação de aspectos relacionados com a fundamentação teórica dos modelos moleculares com granularidade grossa e uso do dielétrico contínuo empregados para estudo do fenômeno da complexação molecular, através da análise, em larga escala, da correlação de cargas em proteínas disponíveis no banco de dados de proteínas. Os estudos teóricos de complexação foram realizados através do emprego de simulações Monte Carlo Metropolis, com o modelo do solvente contínuo, constante dielétrica homogênea e sal implicitamente descrito. Os estudos de complexação permitem compreender os mecanismos moleculares envolvidos, elucidando, inclusive, controvérsias experimentais e oferecendo orientação para aplicações. Já os resultados do estudo de correlação de cargas em estruturas experimentais de proteínas permitem justificar as aproximações do modelo. / The structure and behavior of proteins are of great interest for a number of science fields, with special emphasis to food industry and health. Proteins comprise a fundamental group of biological molecules, with the capacity of interacting with several other structures and creating complexes. Two major topics are covered in this study: (a) evaluation of complex formation among three whey proteins (-lactalbumin, -lactoglobulin and lactoferrin) under different experimental conditions of ionic strength and pH of solution through free energy derivatives and (b) evaluation of features related to the theoretical basis of the coarse-grained molecular models and the use of continuous dielectric which are employed to investigate the molecular complexation phenomenon, through analysis, on a large scale, of charge correlation in proteins available in the protein data bank. The theoretical studies of complexation were performed through the use of Monte Carlo Metropolis simulations, with continuous solvent model, homogeneous dielectric constant and salt implicitly described. The complexation studies allow to understand the molecular mechanisms involved, also elucidating experimental controversies and offering orientation to applications; whereas the results of the charge correlation study in experimental structure of proteins allow justify the approximations of the model.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2018-11-29T17:12:51Z No. of bitstreams: 1 2013 - Renata Silva Cabral de Souza.pdf: 1753830 bytes, checksum: 2dff8752eb3f7974e34f8809d059f126 (MD5) / Made available in DSpace on 2018-11-29T17:12:51Z (GMT). No. of bitstreams: 1 2013 - Renata Silva Cabral de Souza.pdf: 1753830 bytes, checksum: 2dff8752eb3f7974e34f8809d059f126 (MD5) Previous issue date: 2013-05-23 / CAPES / The aim of this study was to evaluate the process of protein concentration in bovine whey proteins by ultrafiltration process and subsequently the protein hydrolysate obtained by enzymatic hydrolysis to produce bioactive peptides with potential antimicrobial and antioxidant activities. For concentration process was used a ceramic ultrafiltration membrane with a molecular range cut-off of 10-20 kDa, transmembrane pressure of 5 bar and, temperature of 30 ?C, 40 ?C and 50 ?C . The optimum temperature condition was at 40 ?C. The Volume Concentrate Factor (VCF) parameter was used as a end-point of the ultrafiltration process and fixed at 2, corresponding on concentrating the initial volume twice, in volume. At the temperature of 40 ?C, VCF had a correspondence on final protein concentration on the concentrated fraction by ultrafiltration and confirmed by Bradford method. Two commercial enzymes were tested Alcalase, Flavourzyme and an equivalent mixture of both 50:50 (w/w) in the hydrolysis reaction. The hydrolysis conditions were determined according to manufacturer instructions and confirmed by other studies: 60 ?C and pH 8 for Alcalase; 50 ?C and pH 7 for Flavourzyme; 50 ?C and pH 8 for enzyme mixture with enzyme / substrate ratio (w / w) 5/100 for all enzymes. The reaction was monitored by pH Stat method. The final Degree of Hydrolysis (DH) achieved was 15%, 52% and 63% for Flavourzyme, Alcalase and enzyme mixture, respectively. Five aliquots were collected along the hydrolysis for each enzyme reaction corresponding to differents DH in order to evaluatethe antioxidant activity by ORAC and ABTS assays with final values between 597- 1092 m? TE (ABTS) and from 1615 to 2920 ?M TE (ORAC) for Flavourzyme; 998-6290 ?M TE (ABTS) and 3092-7567 ?M TE ( ORAC) for Alcalase and finally 913-2678 ?M TE (ABTS) and 2547-5903 ?M TE (ORAC) for the enzyme mixture. The samples from all hydrolysates showed no antimicrobial activity against strains of Salmonella choleraesuis subsp. Enteritidis (ATCC 13076) and Listeria monocytogenes (ATCC 9117). / A proposta do presente trabalho foi avaliar a concentra??o das prote?nas do soro de leite bovino por ultrafiltra??o e posterior obten??o de hidrolisados proteicos deste concentrado via hidr?lise enzim?tica visando obter pept?deos bioativos com potencial atividade antimicrobiana e antioxidante. Para concentra??o das prote?nas do soro foi utilizada membrana cer?mica de ultrafiltra??o com massa molar de corte de 10-20 kDa, press?o aplicada ? membrana de 5 bar, temperaturas testadas (30 ?C, 40 ?C e 50 ?C) . A temperatura ?tima selecionada foi de 40 ?C. O Fator de Concentra??o Volum?trica foi o par?metro utilizado para indicar o final do processo de ultrafiltra??o sendo fixado em duas vezes o volume inicial da alimenta??o. Na temperatura de 40 ?C foi obtida correspond?ncia entre a concentra??o volum?trica e a concentra??o proteica final na fra??o retida pela UF, que tamb?m foi o dobro da encontrada na fra??o alimenta??o, avaliada pelo m?todo de Bradford. Foram testadas duas enzimas comerciais: Alcalase, Flavourzyme e uma mistura equivalente de ambas, na propor??o 50:50 (m/m) na rea??o de hidr?lise. As condi??es de rea??o enzim?tica foram determinadas de acordo com instru??es do fabricante e corroboradas por outros estudos em: 60 ?C, pH 8 para Alcalase; 50 ?C, pH 7 para Flavourzyme; 50 ?C, pH 8 para mistura enzim?tica e rela??o enzima/substrato (g/g) foi de 5/100 para todas as enzimas. A rea??o de hidr?lise foi monitorada pelo m?todo pH Stat. Os Graus de Hidr?lise (GH) finais alcan?ados foram de 15 %, 52 % e 63 % para Flavourzyme, mistura enzim?tica e Alcalase, respectivamente. Foram coletadas cinco al?quotas correspondentes a diferentes GH ao longo da rea??o para cada condi??o enzim?tica utilizada e avaliadas quanto a atividade antioxidante pelos m?todos ABTS e ORAC tendo valores entre 597 a 1092 ?M TE (ABTS) e 1615 a 2920 ?M TE (ORAC) para Flavourzyme, 998 a 6290 ?M TE (ABTS) e 3092 a 7567 ?M TE (ORAC) para Alcalase e por fim, 913 a 2678 ?M TE (ABTS) e 2547 a 5903 ?M TE (ORAC) para a mistura enzim?tica. Nenhuma das amostras de hidrolisado com diferentes GH apresentou atividade antimicrobiana contra cepas de Salmonella choleraesuis subsp. Enteritidis (ATCC 13076) e Listeria monocytogenes (ATCC 9117).

Various methods for extraction and analysis of high value minor proteins (lactoferrin, lactoperoxidase and immunoglobulins) directly from raw milk were explored. Extraction, purification and analysis of high-value minor proteins directly from milk without pre-treatment are major challenges for dairy industry, largely due to the complexity of milk and the presence of colloidal solids (casein micelles and milk fat globules). To overcome some of these challenges, this work focused on three main objectives: 1) characterization of cryogel monolith chromatography for purification of lactoferrin (LF) and lactoperoxidase (LP) directly from raw milk in single step, 2) identification and characterization of Protein A Mimetic affinity ligands for purification of immunoglobulins (Igs) from milk and 3) development and validation of a surface plasmon resonance method for simultaneous quantification of five whey proteins in multiple samples. Results portrayed the possibility of 40–50 column volumes of various milk samples (whole milk, skim milk and acid whey) to pass through a 5 mL cryogel monolith chromatography column at 525 cm hr⁻¹ without exceeding its pressure limits if the processing temperature is maintained around 35–37°C. Ideally, this should be the milk secretion temperature. The dynamic binding capacity obtained for the cryogel matrix (2.1 mg mL⁻¹) was similar to that of the binding capacity (2.01 mg mL⁻¹) at equilibrium with 0.1 mg mL⁻¹ of lactoferrin in the feed samples. Lactoferrin and lactoperoxidase was selectively bound to the cryogel column with trivial leakage in flowthrough fractions. Lactoferrin was recovered from elution fractions with a yield of 85% and a purity of 90%. These results, together with the ease of manufacture, low cost and versatile surface chemistry of cryogels suggest that they may be a good alternative to packed-bed chromatography for direct capture of proteins from milk, provided that the binding capacity can be increased. A Protein A Mimetic (PAM) hexapeptide (HWRGWV) peptide ligand that binds to the Fc portion of antibody molecules was explored for affinity purification of immunoglobulins from milk. The peptide has the ability to purify IgG from various milk and whey samples with a purity of greater than 85% in single step. More than 90% bound IgG was recovered with 0.2 M acetate buffer at pH 4.0 and total column regeneration was successfully achieved by 2.0 M guanidine-HCl. At 9.0 mg mL⁻¹ of IgG feed concentration, an equilibrium binding capacity of 21.7 mg mL⁻¹ and dynamic binding capacity of approximately 12.0 mg mL⁻¹ of resin was obtained. Recoveries and yields of IgG were significantly influenced by the feed IgG concentration. PAM hexamer ligand also contributed a significant amount of cross-reactivity with casein, glycomacropeptides and β-lactoglobulin proteins, however majority of these proteins were recovered in the regeneration step, except β-lactoglobulin, which co-eluted with IgG. Higher IgG concentration in feed vastly reduced the amount of cross-reactivity whilst increasing the recoveries and purities in the final product. PAM affinity ligands also showed interactions towards other classes of bovine immunoglobulins. These findings established the possibility of using PAM hexamer peptide as an alternative to conventional Protein A/G affinity chromatography for the isolation of Igs from milk in single step process. A surface plasmon resonance (SPR) method was developed for simultaneous, quantitative determination of commercially important whey proteins in raw and processed milk samples, whey fractions and various milk-derived products, with six samples per assay. Immobilized antibody stability and reproducibility of analyses were studied over time for 25 independent runs (n=300), giving a relative standard deviation (RSD) of <4%. Immobilized antibodies showed negligible non-specific interactions (<2–4 SPR response units (RU)) and no cross-reactivity towards other milk components (<1 RU). Regeneration of immobilised antibodies with glycine at pH 1.75 was determined to be optimal for maintaining the SPR response between samples. This method compared and validated well with reversed phase high performance liquid chromatography (RP-HPLC) and standard enzyme-linked immunosorbent assays (ELISA).

Orientador: Jaime Amaya-Farfan / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-09T15:48:38Z (GMT). No. of bitstreams: 1 Silva_LucianoBrunodeCarvalho_D.pdf: 1699053 bytes, checksum: 6880af0f88f1ae6dfcbcd9a9e0752ae2 (MD5) Previous issue date: 2008 / Resumo: A Esclerose Lateral Amiotrófica (ELA) é uma doença progressiva neurodegenerativa que envolve, neurônios motores do córtex cerebral, cérebro e medula espinhal. Em aproximadamente 50% dos pacientes, a doença afeta inicialmente os braços. Em 25%, a doença afeta as pernas e, em 25%, o início da doença é bulbar. Ocorre desnutrição em 16%- 50% dos pacientes com ELA, principalmente pela disfunção da deglutição. Para 50% dos casos, a ELA é comumente fatal dentro de 3-4 anos a partir da ocorrência de pneumonia. Muitas complicações nutricionais que acontecem com o paciente com ELA, geralmente se devem à viscosidade da dieta. Os líquidos de baixa viscosidade dificultam a deglutição de pacientes que apresentam controle laríngeo reduzido. O objetivo do presente trabalho foi diagnosticar o estado nutricional, bem como avaliar o efeito da suplementação com isolado protéico do soro de leite aglomerado com polissacarídeo no estado nutricional e condições gerais de pacientes com ELA. A investigação da ingestão alimentar se deu por meio da avaliação do consumo alimentar, qualitativo e quantitativo, de pacientes com ELA. Aplicou-se o questionário de freqüência de consumo alimentar e o recordatório de 24 horas. Todos os pacientes apresentaram inadequação para energia, fibras, cálcio e vitamina E. Considerando-se a predominância topográfica do comprometimento muscular na doença, foram observadas diferenças significativas entre pacientes predominantemente bulbares e de predomínio apendicular. Nos pacientes com maior envolvimento apendicular (GA) houve maior ingestão energética (p=0,02), de gordura saturada (p=0,03), monoinsaturada (p=0,04) e polinsaturada (p=0,001), além de colesterol (p=0,001) e fibras (p=0,001). Nos pacientes com predominância bulbar (GB) houve dificuldade no manejo da consistência da dieta. A partir daí, foram elaborados suplementos a base de proteínas do soro de leite e polissacarídeo. O aglomerado contendo 70% de isolado protéico do soro de leite e 30% de amido modificado (70%WPI:30%AM) na concentração de 28g em 250mL em todos os sistemas avaliados, suco de laranja e leite integral, exibiu viscosidade entre 2122,66 e 5110,60 centipoises, além de maior nota, nos testes sensoriais, 6,97 (± 0,15), quando comparado aos outros produtos, não diferindo (p=0,681) do espessante comercial, que obteve nota 6,91 (± 0,38). Baseando-se nestes resultados, sugeriu-se a utilização do aglomerado 70%WPI:30%AM para terapia nutricional de pacientes com ELA. Previamente à administração do suplemento, em função dos poucos dados sobre as técnicas de avaliação nutricional em ELA, foram correlacionados os indicadores mais comuns na avaliação nutricional, funcional e respiratória dos pacientes. No GA, observou-se correlação entre: Pressão inspiratória máxima (Pimax) e expiratória máxima (Pemax) (r=-0,76); Pemax e oximetria de pulso (r=0,58); Pimax e porcentagem de perda de peso (%PP) (r=0,59); e entre Pimax, escore total e subescala respiratória (ALSFRS) com %PP (r=0,59). No GB, houve correlação entre Pemáx e IMC (r=0,97). Em ambos GA e GB, observaram-se correlação entre índice de massa corporal (IMC) e as variáveis: massa (kg), gordura (%), circunferência braquial e punho (cm), pregas cutâneas tricipital, subescapular e supra-ilíaca (mm), circunferência muscular do braço (cm), área muscular e gordurosa do braço (mm2). Definidas as técnicas de avaliação, 16 pacientes foram divididos em dois grupos: suplementado (70%WPI:30%AM) e controle (maltodextrina) e submetidos a avaliação nutricional, respiratória e funcional durante 4 meses. No grupo suplementado verificou-se melhora do estado nutricional geral, aumento do peso e Índice de Massa Corporal-IMC, circunferência e área muscular do braço (CMB e AMB), albumina, linfócitos e leucócitos, diminuindo os marcadores do catabolismo muscular, creatina quinase, aspartato-aminotransferase e alanina-amino-transferase. No grupo controle, observou-se diminuição do peso, IMC, CMB e AMB e aumento da área gordurosa do braço, sem alterações significativas nos parâmetros bioquímicos. O presente estudo mostrou que o suplemento 70%WPI:30%AM melhorou o estado nutricional e condições gerais pacientes com ELA, podendo ser utilizado na prática clínica / Abstract: Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder that involves, motor neurons in the cerebral cortex, brainstem, and spinal cord. In approximately 50% of the patients, the disease first affects the arms, in 25% the legs and in 25% the onset is bulbar. Malnutrition occurs in 16%-50% of patients with ALS, mainly for swallowing difficulty. For 50% of the cases, ALS is frequently fatal inside of 3-4 years from the pneumonia occurrence. Many of the nutritional complications that occur with ALS sufferers are due to the low viscosity of the diet. Fluid liquids are difficult to swallow by patients with reduced deglutition control. The objective of the present work was to diagnosis the nutricional state as well as evaluating the effect of the supplementation with milk whey protein isolate agglomerated with modified starch in the nutricional state and general conditions of patients with ALS. The investigation of the alimentary intake was evaluate quantitatively and qualitatively food intake in patients with ALS. We employed the frequency questionnaire and the 24h-diet history. All patients presented inadequate energy, fibers, calcium and vitamin E intake. Taking into account predominantly affected muscle groups, we found significant differences between patients with bulbar or appendicular onset. Patients with appendicular ALS had higher intake of energy (p=0.02), saturated fat (p=0.03), monounsaturated fat (p=0.04), polyunsaturated fat (p=0.001), as well as cholesterol (p=0.001) and fibers (p=0.001). In the patients with predominance to bulbar (GB) had difficulty in the handling of the consistency of the diet. From there, supplements had been elaborated from protein base of whey milk and modified starch. The agglomerate contend 70% whey protein isolate and 30% modified starch (70%WPI:30%MS), in all the systems evaluated, at a concentration of 112g/L showed a viscosity between 2,122 and 5,110cP. In the sensory tests, the 70%WPI:30%MS in orange juice obtained the highest score, 6.97 (± 0.15), in relation to the other agglomerates, not differing (p=0.681) from the commercial thickener, 6.91 (p=0.380). Based on these results, the agglomerate 70%WPI:30%MS was suggested for use in the nutritional therapy of patients with ALS. Previously the administration of the supplement, in function of the few data on the techniques of nutricional evaluation in ALS, had been correlated the pointers most common in the nutricional, functional and respiratory evaluation of the patients. Among the GA type significant correlation was observed between maximal inspiratory (MIP) and expiratory (MEP) pressure (r=-0.76), MEP and pulse oxymetry (r=0.58), MIP and percent weight loss (%WL; r=0.59), and between MIP, total and subscale respiratory scores (ALSFRS-R) with %WL. With regard to the GB, correlation was found between MEP and BMI (r=0.97). In both GA and GB correlations were noticed between the BMI and the variables mass (kg), fat (%), arm and fist circumference (cm), and tricipital, subscapular and supra-iliac skinfolds (mm), as well as the arm muscle circumference (cm) and fatty arm muscular area (mm2). After defined the evaluation techniques, 16 patients were randomized to two groups, treatment (70%WPI:30%MS) and placebo (maltodextrin). They underwent prospective nutritional, respiratory and functional assessment for 4 months. Patients in the treatment group presented weight gain, increased BMI, increased arm muscle area and circumference, higher albumin, white blood cell and total lymphocyte counts, and reduced markers of skeletal muscle turnover (creatine-kinase, aspartate transferase and alanine transferase). In the placebo group, biochemical measures did not change, but weight and BMI declined. Our results indicate that the agglomerate 70%WPI:30%MS may be useful in the nutritional therapy of patients with ALS / Doutorado / Nutrição Experimental e Aplicada à Tecnologia de Alimentos / Doutor em Alimentos e Nutrição

Made available in DSpace on 2014-07-29T16:16:35Z (GMT). No. of bitstreams: 1 Dissertacao Dayana Silva Batista Soares.pdf: 1898125 bytes, checksum: 54ca2333fb4f6f671b7ff76b9d77c10d (MD5) Previous issue date: 2012-02-24 / The packaging has as main function to maintain the integrity of the product, protecting it against mechanical damage, moisture loss and gain. Most packaging is manufactured from non-biodegradable materials and non-renewable source promoting great environmental impacts. Among the raw materials used in the development of biodegradable films found the whey proteins. The biodegradables films do not present the same resistance presented for the synthetic films, many treatments are being applied to biofilmes in the attempt to improve the mechanical properties, amongst them the variation of pH and irradiation. In this way, aimed to evaluate the effect of different pH values and doses of gamma radiation on the mechanical, optical and barrier properties of whey proteins biodegradable films. The films were made by casting and characterized with respect to thickness, color, opacity, water vapor permeability, water solubility, scanning electron microscopy and mechanical properties. The different treatments studied significantly influenced the properties of the films, except for the water solubility. It is concluded that the different treatments can promote improvement in film properties. / A embalagem tem como função principal manter a integridade do produto, protegendo-o contra danos mecânicos, perda e ganho de umidade. A maior parte das embalagens é fabricada com materiais não biodegradáveis e de fonte não renovável promovendo grande impacto ambiental. Dentre as matérias-primas empregadas na elaboração de filmes biodegradáveis encontramos as proteínas do soro de leite. Os filmes biodegradáveis não apresentam a mesma resistência apresentada pelos filmes sintéticos logo, muitos tratamentos estão sendo aplicados aos filmes biodegradáveis na tentativa de melhorar as propriedades mecânicas, dentre elas a variação de pH e irradiação. Desta maneira, objetivou-se avaliar o efeito de diferentes valores de pH e doses de radiação gama nas propriedades mecânicas, ópticas e de barreira de filmes biodegradáveis de proteínas do soro de leite. Os filmes foram elaborados pelo método de casting, e caracterizados quanto à espessura, cor, opacidade, permeabilidade a vapor de água, solubilidade em água, microscopia eletrônica de varredura e propriedades mecânicas. Os diferentes tratamentos estudados influenciaram significativamente as propriedades dos filmes, com exceção à solubilidade em água. Concluise que os diferentes tratamentos empregados podem promover melhora nas propriedades dos filmes.

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.