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PONTE, Paulo Roberto Lins. Avaliação do teste químico no diagnóstico de intolerância à lactose e sua associação com polimorfismo genético em uma população de Fortaleza/CE. 2012. 123 f. Dissertação (Mestrado em Farmacologia) - Universidade Federal do Ceará. Faculdade de Medicina, Fortaleza, 2012. / Submitted by denise santos (denise.santos@ufc.br) on 2013-05-09T16:15:37Z No. of bitstreams: 1 2012_dis_prlpontes.pdf: 583734 bytes, checksum: 373826bf1b841d5e85edbf60a758f624 (MD5) / Approved for entry into archive by Erika Fernandes(erikaleitefernandes@gmail.com) on 2013-05-16T13:41:50Z (GMT) No. of bitstreams: 1 2012_dis_prlpontes.pdf: 583734 bytes, checksum: 373826bf1b841d5e85edbf60a758f624 (MD5) / Made available in DSpace on 2013-05-16T13:41:50Z (GMT). No. of bitstreams: 1 2012_dis_prlpontes.pdf: 583734 bytes, checksum: 373826bf1b841d5e85edbf60a758f624 (MD5) Previous issue date: 2012 / Intolerance to lactose is a clinical syndrome comprising one or more corteges symptomatology associated with the ingestion of milk (clinical phenotype intolerant) and lactose tolerance in the absence of symptoms (clinical phenotype tolerant). Thus, in practice, must be attentive to cases similar to these badly absorptive symptoms because the convergences and differences between these disorders are tenuous and diagnoses are often inaccurate, making treatment and unsatisfactory conduct. Therefore, studies are needed using different diagnostic methods, and compare the different methods used in clinical practice and determine the most appropriate to give a conclusive diagnosis, since it is not always easy, given that the symptoms suggest other diseases. Objective.To evaluate the phenotypic characteristics, the response to chemical test for lactose tolerance and the genotype of patients with diagnostic hypothesis of intolerance to lactose and the correlation between them.Material and methods. We conducted a cross-sectional study at the University Hospital Walter Cantidio (HUWC), with 173 patients who had positive phenotype and were seen at the outpatient service of servers HUWC, from January to August 2010. The final sample consisted of 119 patients who underwent the lactose tolerance test and genotyping of the gene for lactase-phlorizin hydrolase (LPH). Results. Among the participants, most were: female, 54.6% (65 cases), the age group of 46-55 years old, 26.5% (32 cases); married, 65.6% (78 cases); dun-colored, 53.8% (65 cases); higher level, 42.9% (51 cases). The prevalence of intolerance to lactose was 45.4% and chemical intolerance was 73.9%. Symptoms investigated more prevalent in lactose intolerant were flatulence 81.4% (44 cases); bloating, 68.5% (37casos); borborygmus, 59.3% (32 cases) and diarrhea, 46, 3% (25 cases). The presence of the polymorphisms C> T polymorphisms and 13 910-G> A-22018 in LPH gene was identified in individuals selfreferred as lactose intolerant, and is therefore an excellent test for the diagnosis. The diagnosis of lactose intolerance that most had adequate baseline the difference was <15 with ROC curve greater than 80.3%, with good sensitivity and specificity. Conclusions. There is a correlation between genotype and chemical intolerant individuals, suggesting that the molecular test could be proposed for the laboratory diagnosis of lactose intolerance, and the chemical test, the difference basal <15 was more appropriate. / A intolerância à lactose é uma síndrome clínica composta por um ou mais cortejos sintomatológicos associados à ingestão de lácteos (fenótipo clínico intolerante) e tolerância à lactose na ausência desses sintomas (fenótipo clínico tolerante). Assim, na prática clínica, deve-se estar atento aos casos semelhantes a essa sintomatologia disabsortiva, pois as convergências e diferenças entre esses distúrbios são tênues e os diagnósticos muitas vezes são imprecisos, tornando o tratamento e a conduta insatisfatórios. Portanto, são necessários estudos que utilizem diferentes métodos diagnósticos, assim como comparar os diferentes métodos utilizados na prática clínica e averiguar o mais apropriado para dar o diagnóstico conclusivo, uma vez que nem sempre é fácil, haja vista que os sintomas sugerem outras doenças. Objetivo. Avaliar as características fenotípicas, a resposta ao teste químico de tolerância à lactose e genótipo de pacientes com hipótese diagnóstica de intolerância à lactose e a correlação entre eles. Material e métodos. Foi realizado um estudo transversal no Hospital Universitário Walter Cantídio (HUWC), com 173 pacientes que apresentavam fenótipo positivo e foram atendidos no serviço do ambulatório dos servidores do HUWC, no período de janeiro a agosto de 2010. A amostra final foi composta por 119 pacientes que realizaram o teste de tolerância à lactose e genotipagem do gene da florizina-lactase hidrolase (LPH). Resultados. Dentre os participantes, a maioria era: do sexo feminino, 54,6% (65 casos); da faixa etária de 46-55 anos, 26,5% (32 casos); casada, 65,6% (78 casos); de cor parda, 53,8% (65 casos); de nível superior, 42,9% (51 casos). A prevalência de intolerância à lactose foi de 45,4% e a de intolerância química foi de 73,9%. Os sintomas investigados que apresentaram maior prevalência no intolerante à lactose foram: flatulência, 81,4% (44 casos); empachamento, 68,5% (37casos); borborigmo, 59,3% (32 casos); e diarreia, 46,3% (25 casos). A presença dospolimorfismos C>T-13910 e polimorfismos G>A-22018 no gene da LPH foi identificada nos indivíduos autorreferidos como intolerantes à lactose, sendo, portanto, um excelente exame para o diagnóstico. O diagnóstico de intolerância à lactose que mais se apresentou adequado foi a diferença basal <15 com a curva de ROC maior que 80,3%, com boa sensibilidade e especificidade. Conclusões. Observa-se uma correlação genotípica e química entre os indivíduos intolerantes, sugerindo que o teste molecular poderia ser proposto para o diagnóstico laboratorial de intolerância à lactose, e, no teste químico, a diferença basal <15 foi a mais adequada.

Orientadores: Valdemiro Carlos Sgarbieri, Alda Luiza Santos Lerayer / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-02T22:36:22Z (GMT). No. of bitstreams: 1 Saron_MargarethLopesGalvao_M.pdf: 23626715 bytes, checksum: 97334ba53a2521a3bbac09b596a00d1f (MD5) Previous issue date: 2003 / Resumo: O presente trabalho teve por objetivo estudar novas aplicações para o permeado proveniente da concentração das proteínas do soro de leite bovino pelo processo de ultrafiltração em membrana (PM de corte de 10 Kda). A concentração do permeado, evaporação sob vácuo permitiu a insolubilização da lactose e sua purificação. Através de reação de isomerização em pH alcalino e o uso de borato de sódio como catalizador, obteve-se a conversão da lactose em lactulose e sua purificação por cromatografia de troca iônica e exclusão molecular em colunas. A lactulose purificada foi testada in vitro como prebiótico em vários meios de cultura complementados com o permeado de soro de leite, para o crescimento das bactérias probióticas (Lactobacillus acidophilus La-5 e Bifidobacterium /actis Bb-12). Verificou-se que tanto o permeado de soro de leite quanto a lactulose funcionaram como importantes fatores de crescimento para essas bactérias. O permeado de soro contem aproximadamente 85% de lactose (em base seca), sendo bastante deficiente em aminoácidos. Após separação da lactose, o líquido sobrenadante do permeado representa uma boa fonte de vitaminas, particularmente B1, B2 e niacina. A cromatografia em camada delgada (CCO) foi eficiente para a detecção da lactulose após eluição em colunas e, através da cromatografia líquida de alta eficiência (CLAE) pode-se confirmar a reação de isomerização da lactose em lactulose com rendimento de aproximadamente 70%. Os ensaios microbiológicos mostraram que a complementação do meio MRS (De Man, Rogosa e Sharpe) com permeado de soro de leite (PSL) promoveu um aumento significativo na produção de células de L.acidophi/us e B./actis. O PSL, quando combinado com extrato de levedura (EXT) na proporção de 75/25, mostrou-se adequado para a propagação de células de B. /actis. O meio MRS adicionado de 2% ou 3% de lactulose apresentou um aumento do crescimento de L. acidophi/us. A concentração de apenas 1% de factulose no meio MRS modificado e PSUEXT foi eficiente para estimular o rescimento de Bifidobacterium /actis. Tanto o L. acidophilus como a B. /actis demonstraram resistência as condições de acidez (pH 2 , 3h, 37°C). A presença de lactulose no meio de cultura promoveu maior proteção destas bactérias quando expostas à presença de sais biliares. Neste trabalho são descritos ainda aspectos relevantes dos efeitos benéficos dos prebióticos e probióticos para a saúde humana. / Abstract: The objetive of the present work was to study new application for the permeate obtained during concentration of bovine milk whey by the process of membrane ultrafiltration (MW cut off 10 Kda). The concentration of the permeate, by vacuum evaporation, permited the insolubilization and further purification of the lactose by crystallization. By an isomerization reaction at alkaline pH using sodium borate as catalyst, the conversion of lactose to lactulose was promoted which was purified in ionic exchange and molecular exclusion colunn chromatography. The purified lactulose was tested in vivo as prebiotic in various culture media complemented whith whole whey permeate, for the growth of the probiotic bacteria Lactobacillus acidophilus La-5 and Bifidobacterium lactis Bb-12. It was verified that both the whole permeate as well as the lactulose functioned as important growth promoting factors for these bacteria. The whey permeate contained approximately 85% lactose on a dry basis and was deficient in amino acids. After lactose separation the permeate supernatant liquid contained appreciable quantities of vitamins, particularly B1, B2 and niacin. Thin layer chromatography (TLC) was efficient in the detection of lactulose after elution from the columns, and. high perfomance liquid chromatography (HPLC) was used to confirm the isomerization reaction of lactose to lactulose whith a yield of approximately 70%. The microbiological assay showed that complementation of the MRS (De Man, Rogosa and Sharpe) medium whith whole milk whey permeate significant increase in cell production of both L. acidophilus and B. lactis. The modified MRS medium (MRSm) added of 2% or 3% lactulose showed an increase in growth for the L. acidophilus. At only 1% lactulose concentration in the MRSm or MWPNE media a significant increase in growth was observed for B. lactis. Both L. acidophilus and B. lactis showed resistance to acidity condition (pH 2, 3h, 37°C). The presence of lactulose in the culture media promoted higher protection to these bacteria when exposed to biliary salts. In this work important aspects of the beneficial effects of prebiotics and probiotics to human health were described. / Mestrado / Mestre em Alimentos e Nutrição

Orientador: Edgard Ferro Collares / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-07-20T18:08:46Z (GMT). No. of bitstreams: 1 Machado_FernandodeAlmeida_D.pdf: 2699170 bytes, checksum: c4091cce3dcc6e6257e79337cec38003 (MD5) Previous issue date: 1995 / Resumo: O presente trabalho teve como objetivo, numa primeira etapa, verificar, em ratos adultos, as retenções gástricas (RG) de soluções aqüosas de lactose, sacarose e maltose, todas na concentração a 10% (p/v), e a influência da associação combinada entre dois desses açúcares sobre o esvaziamento gástrico e, numa segunda etapa, verificar se essa influência se mantém quando se adiciona sacarose ou maltose ao leite de vaca integral. Foram utilizados 120 ratos Wistar machos com idade entre 8 a 10 semanas, que receberam suas respectivas refeições de prova por via oro gástrica, através de uma sonda metálica, no volume de 2 ml/l00 g de peso. As soluções de açúcares foram marcadas com fenol vermelho (6 mg/dl) e os leites foram marcados com PEG 4000 (2 g/dl). As RG foram determinadas calculando-se a quantidade de marcado retido no estômago, após leitura espectrofotométrica. Na primeira etapa do experimento, foram utilizados 48 animais, divididos eqüitativamente em 6 subgrupos, de acordo com a refeição de prova utilizada: lactose 10% (L), sacarose 10% (S), maltose 10% (M), lactose 5% + sacarose 5% (LS), lactose 5% + , maltose 5% (LM) e sacarose 5% + maltose 5% (SM) (p/v). A RG foi avaliada após 15 minutos da infusão orogástrica da refeição de prova. Na segunda etapa do estudo, foram utilizados 72 animais divididos em 3 subgrupos eqüitativos, de acordo com a refeição-de prova: leite de vaca integral sem acréscimo de açúcar, leite de vaca com sacarose 5% (p/v) e leite de vaca com maltose 5% (p/v). Para cada refeição de prova, foi avaliada a RG ao tempo de 15, 30 e 45 minutos, utilizando-se, em cada momento, 8 animais em cada subgrupo. Foram utilizados os testes estatísticos não paramétricos de Kruskal- W allis, com níveis de significância de 10%. Em seguida, foram feitos os testes de comparações múltiplas, com níveis de significância de 1% e 2%, respectivamente. Os resultados do estudo mostram que são significativas as diferenças entre as RG das soluções de L e M, SeM, S e SM, L e LM, L e SM. A maltose, associada à lactose ou à sacarose, promoveu retenção gástrica significativamente maior que a de solução de lactose ou sacarose isoladas, mantendo-se a densidade energética / Abstract: This study has the goals of establishing the gastric retentions of aqueous solutions of lactose, sucrose and maltose, all of them at 10% (w/v), and also the influence of associations between two of these sugars on the gastric retentions. Later on, it was studied this influence on gastric retention using sucrose or maltose on whole cow's milk. It was used 120 Wistar male rats, aged from 8 to 10 weeks. The rats received a test meal (2 rnl/l00g weight) with a marker by orogastric route through a metal tube. The markers used were phenol red (6 mg/dl) in the sugars aqueous solutions and polyethylene glycol (PEG) 4000 (2 g/dl) in the cow's milk test meal. The gastric retention was measured by determining the amount ofthe marker staying in the stomach by spectrophotometry. During the first phase of the study, 48 rats were distributed in 6 subgroups, each of them received test meal of aqueous solution of lactose 10% (L), sucrose 10% (S), maltose 10% (M), lactose 5% + sucrose 5% (LS), lactose 5% + maltose 5% (LM) and sucrose 5% + maltose 5% (SM) (w/v). The gastric retentions were determined 15 minutes after the infusion of test meal. In the second phase it was used 72 rats which were equaly distributed in 3 subgroups, according to the test meal: whole cow's milk without sugar addition, cow's milk plus suciose 5% (w/v) and cow's milk plus maltose 5% (w/v). The determinations of gastric retentions were performed at 15, 30 and 45 minutes after the orogastric infusion, being 8 animals used for each time and each test meal. In both phases of the study it was utilized the Kruskal- Wallis test, being a. = 10%. Later, the multiple comparisions were calculated with a. = 1 % and 2%, respectively, to first and second phases. There are significant differences between gastric retention of aqueous solutions of lactose and maltose (L x M), sucrose and maltose (S x M), sucrose and sucrose + maltose (S x SM), lactose and lactose + maltose (L x LM), lactose and sucrose + maltose (L x SM). The gastric retention of maltose plus lactose or maltose plus sucrose was significantly larger than those oflactose or sucrose at the same energetic density / Doutorado / Doutor em Saude da Criança e do Adolescente

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At the last time, we can see increased interest of lactose-free dairy products. The aim of the thesis was to evaluate offer of such products at selected retail chains in the South Bohemian region, then to conclude a sensory assessment of selected products and to verify of public awareness about lactose intolerance with a questionnaire survey. The widest offer of lactose-free dairy products has been found in Globus and Tesco, in the opposite the lowest offer has been found in Billa. Location of these products within the store was most the transparent in Globus (compared other stores). Three samples of white lactose free yoghurts and two samples of milk (with lactose and lactose-free) were evaluated by a sensory assessment. The sample of yoghurt from the company Madeta, line Nature was rated the best. On the contrary sample of yoghurt from the company Tatranská mliekáreň, line Nature´s Promise was rated the worst. The preference between two milk samples among the evaluators was balanced, despite large to moderate differences of intensity were described between them. Most respondents (79 %) knew what the term lactose intolerance means, but substantial part (22 %) of respondents badly indicated this the issue as allergy to milk casein. Because of high number of lactose-intolerant people in population this issue is still very actual, and it needs to be addressed.

Freeze-drying is the process of removal of water or organic solvent from a desired product by means of sublimation at a low temperature and low pressure. It is commonly employed for drying samples which are heat labile and require sensitive treatment, and is mainly used in the pharmaceutical and food industries. It is an expensive process, requiring vacuum, refrigeration and long cycle times, but does yield quality benefits due to the low temperatures involved and the porous nature of the product. Reducing drying times is important to manufacturers, and this depends on optimising rates of heat and mass transfer in the system without the sample losing its porous structure. However, freeze drying is difficult to study experimentally due to the low temperatures and pressures involved. The quality of the final product mainly depends on the sublimation rate and an optimum lyophilisation requires identification of the parameters which influence the process. The main aim of this study is to employ freeze drying microscopy (FDM) as a useful tool to identify these process parameters and help optimise primary drying phase of the freeze drying process for two systems: lactose (relevant to pharmaceuticals) and coffee (the most widely freeze-dried food product). This equipment allows the movement of sublimation fronts to be directly visualised in-situ under carefully controlled (and isothermal conditions), but has scarcely been used in the literature for this purpose. An image analysis method is developed to automatically track the movement of sublimation fronts, and the frontal data fitted to a simple mass transfer model employing surface and bulk resistances. Initial experiments with lactose solution show poor reproducibility in nucleation temperatures during the freezing step and thus primary drying rates. To improve reproducibility, a small amount of silver iodide (AgI) was added to samples which acts as a nucleating agent and increases the nucleation temperature. This addition of AgI also increases the mean ice crystal size in the samples and are easily visible under the freeze-drying microscope, and in many cases show a distinct orientation with respect to direction of sublimation front. Furthermore, the orientation greatly influences sublimation rates, being approximately factor of two faster when crystals are oriented in the direction of mass transfer. FDM experiments with coffee were less straightforward as nucleation temperatures could not be reliably controlled, even with AgI added. Nevertheless there was a clear decrease of bulk resistance with increasing nucleation temperature. An experimental programme was then undertaken to examine the impact of initial solid content, cooling rate, the addition of an annealing step, freeze drying temperature and aeration (for coffee samples). Frontal data were fitted to a simple mass transfer model comprising surface and bulk (per unit depth) resistances and good fits to data were obtained. FDM experiments with lactose and coffee clearly showed the presence of a surface resistance which could also be seen as a surface layer which was devoid of ice crystals (and hence not porous when sublimed). The edge resistance first increased and then decreased with solids content. The resistance per unit depth increased exponentially with solids content, so much so that there is an optimal solids content (around 10% solids) in relation of the rate of production of dried material. Cooling rates were mainly found to affect the surface resistance rather than bulk resistance and this may be due to different levels of surface drying when the samples are being cooled for different lengths of time. Annealing substantially changed the ice crystal sizes, and had a beneficial effect on freeze drying rates and had a similar effect to adding AgI. Freeze drying rates also increased with increasing temperature approximately in line with the saturated vapour pressure (SVP) of ice which is widely held to constitute the driving force for mass transfer. It was possible to make drying time calculations for conventional vial (lactose) and tray (coffee) drying using the frontal rate data obtained from FDM. For 10% lactose and 10% coffee (annealed) there was good agreement between the vial and tray data and predictions based on a microstructure oriented parallel to the direction of mass transfer. This was the only case where agreement was found, but also the only case where directionality was observed in FDM. The much faster drying times observed in the vial and tray experiments are thus attributed to directional solidification occurring in these systems, and this was borne out by SEM imaging. Aeration of the coffee samples was also found to substantially reduce drying times. The influence of microstructure on freeze drying rates is thus very clear.