268 resultados para Gluten


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A Doença Celíaca (DC) é uma doença autoimune que afeta o intestino delgado de indivíduos geneticamente susceptíveis após contato com o glúten. Diversos estudos têm relatado aumento da prevalência ao longo dos anos. Objetivo: Determinar a prevalência de DC em pacientes adultos sem diarreia encaminhados à Disciplina de Gastroenterologia do HUPE da UERJ para serem submetidos a Endoscopia Digestiva Alta (EDA).Comparar os resultados do histopatológico das biópsias duodenais com os resultados sorológicos, utilizando o anticorpo antitransglutaminase tecidual IgA (ATGt IgA). Métodos: Pacientes que foram encaminhados ao nosso serviço para serem submetidos a EDA entre Julho de 2008 e Julho de 2010, com idade entre 18 e 85 anos foram aceitos no estudo. Critérios de exclusão foram cirrose, neoplasias do trato gastrointestinal, HIV, uso de imunossupressores e anticoagulantes, diarreia, hemorragia digestiva e DC. Coleta de sangue para pesquisa do anticorpo ATGt IgA (utilizando KIT ORGENTEC - Alemanha), avaliação endoscópica e exame histopatológico das biópsias de segunda porção duodenal foram feitos para cada paciente. Biópsias foram avaliadas de acordo com o critério de Marsh modificado. Resultados: Trezentos e noventa e nove pacientes consecutivos (112 homens, 287 mulheres), média de idade 49,616,4 anos, variando de 18-85 anos, sem diarreia, foram prospectivamente aceitos. Os sintomas clínicos mais prevalentes foram dor abdominal em 99,5%, pirose em 41,1%, plenitude pós prandial em 30,6%, náuseas e vômitos em 21,3%. Os achados endoscópicos foram: normais em 41,6%, lesões pépticas (esofagite, gastrite, duodenite e úlceras) em 41,6%, hérnia hiatal em 5,5%, pólipos gástricos em 3%, neoplasias em 1,3% e miscelânea em 7%. DC foi endoscopicamente diagnosticada em 13 pacientes (3,3%) com mucosa duodenal exibindo serrilhamento das pregas em 8 (2%), diminuição do pregueado em 2 (0,5%) e mucosa exibindo padrão nodular e mosaico em 3 (0,75%). Os achados histopatológicos de duodeno foram normais em 96,7%, duodenites inespecíficas em 2,7% e 3 pacientes (0,75%) confirmaram DC pelos critérios de Marsh modificado (IIIa, IIIb e IIIc). O anticorpo ATGt IgA foi positivo (>10 U/ml) em 1,3% (5/399). Conclusão: Este estudo mostrou que a prevalência de DC em pacientes dispépticos sem diarreia atendidos na Disciplina de Gastroenterologia e Endoscopia do HUPE/UERJ foi de 0,75% (1:133). A acurácia diagnóstica do anticorpo ATGt IgA é boa para pacientes com Marsh III e achados endoscópicos sugestivos. Nenhum dos pacientes tinha alterações Marsh I ou II. A EDA se mostrou um excelente método de triagem para definir os pacientes com graus mais acentuados de atrofia e que se beneficiariam de biópsia e sorologia para confirmação diagnóstica. Os resultados obtidos neste trabalho não justificam uma triagem rotineira de DC.

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The purpose of the present work was both to analyze composition of Spanish celiac women and to study the food habits and gluten-free diet of these celiac patients, in order to determine whether they achieve a balanced and healthy diet as well as to highlight nutritional qualitative and/or quantitative differences. 54 adult celiac women (34 +/- 13 years) took part in the six-month study. Height, weight and body composition were measured. An analysis of energy consumption and of the macronutrient distribution of their diet was carried out. Their fulfillment of micronutrient intake recommendations was verified. Participants showed a Body Mass Index of 21.6 +/- 2.4 kg/m(2). Energy Intake was slightly lower than the Dietary Reference Intakes. Excessive protein apart from over-consumption of fat was observed. More than three quarters of participants consumed meat in excess. Carbohydrate consumption along with that of fiber was below recommended levels. Vitamin D, iron, and iodine had a low percentage of recommendation compliance. In general, participants followed the recommendations of dairy products and fruit intake whereas vegetable consumption was not enough for the vast majority. We conclude that although the diet of celiac women does not differ much from the diet of general population, some considerations, such as reducing fat and protein consumption and increasing fiber intake, must be taken into account.

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Objective The protein Hwp1, expressed on the pathogenic phase of Candida albicans, presents sequence analogy with the gluten protein gliadin and is also a substrate for transglutaminase. This had led to the suggestion that C. albicans infection (CI) may be a triggering factor for Celiac disease (CeD) onset. We investigated cross-immune reactivity between CeD and CI. Methods Serum IgG levels against recombinant Hwp1 and serological markers of CeD were measured in 87 CeD patients, 41 CI patients, and 98 healthy controls (HC). IgA and IgG were also measured in 20 individuals from each of these groups using microchips sensitized with 38 peptides designed from the N-terminal of Hwp1. Results CI and CeD patients had higher levels of anti-Hwp1 (p= 0.0005 and p= 0.004) and anti-gliadin (p= 0.002 and p= 0.0009) antibodies than HC but there was no significant difference between CeD and CI patients. CeD and CI patients had higher levels of anti-transglutaminase IgA than HC (p= 0.0001 and p= 0.0039). During CI, the increase in anti-Hwp1 paralleled the increase in anti-gliadin antibodies. Microchip analysis showed that CeD patients were more reactive against some Hwp1 peptides than CI patients, and that some deamidated peptides were more reactive than their native analogs. Binding of IgG from CeD patients to Hwp1 peptides was inhibited by gamma III gliadin peptides. Conclusions Humoral cross-reactivity between Hwp1 and gliadin was observed during CeD and CI. Increased reactivity to Hwp1 deamidated peptide suggests that transglutaminase is involved in this interplay. These results support the hypothesis that CI may trigger CeD onset in genetically-susceptible individuals.

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[ES] La enfermedad celíaca (EC) es una enteropatía autoinmune de predisposición genética, producida por la ingestión en la dieta de péptidos derivados de cereales como el trigo o la cebada. Aunque se creía que afectaba casi de forma exclusiva a los individuos europeos (1%), actualmente se conocen casos en todo el mundo. El modelo patogénico se centra en los mecanismos de la inmunidad adaptativa dependientes de la estimulación de linfocitos T CD4+ reactivos, pero existe además un efecto tóxico directo del gluten sobre el epitelio intestinal, dependiente de la inmunidad innata. La participación de la Genética en la susceptibilidad a la enfermedad es conocida desde hace tiempo, siendo el locus HLA el que explica aproximadamente el 40% del componente genético de la enfermedad. Para tratar de identificar otros genes con susceptibilidad, se han venido realizando múltiples esfuerzos durante los últimos años. Uno de los últimos, llevado a cabo en 2011, fue el Proyecto Immunochip. En él, se analizaron más de 200.000 variantes y se descubrieron 13 nuevos loci de riesgo para la EC, que junto con los descubiertos en anteriores trabajos y el locus HLA, daban un total de 40 loci de riesgo. Entre ellos, se encontraba la región que ocupa el gen LPP . Localizado en el cromosoma 3, un estudio reciente lo vincula con los procesos de adhesión celular en el intestino. En el presente trabajo, se ha estudiado el efecto de la gliadina sobre la expresión del gen de interés (LPP ) y el posible efecto de un silenciamiento del mismo sobre dos genes relacionados con las uniones celulares (ACTB y TJP1). En el caso de la gliadina, no se halló un cambio significativo en la expresión del gen. Mientras, los resultados del efecto del silenciamiento fueron dispares, no siendo concluyentes para el gen ACTB, pero encontrando una posible asociación entre los genes LPP y TJP1.

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La enfermedad celíaca, también conocida como enteropatía sensible al gluten, es un trastorno autoinmune crónico causada por la intolerancia a la ingesta de gluten y desarrollada por personas genéticamente predispuestas cuyo único tratamiento consiste en una dieta estricta libre de gluten. El desarrollo de dicha enfermedad está relacionada con una gran variedad de genes entre los que se encuentra PTPRK. Este trabajo desarrolla un estudio de la expresión de dicho gen así como un protocolo de silenciamiento del mismo.

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Changes in the texture (elastic nature) of the flesh of barrel salted herring during the ripening process at 4°C have been monitored. The method employs the analysis of stress-relaxation curves after compression to half of the sample thickness on an lnstron Model 1112. The parameter 'T/P' for each sample represents the reciprocal of the gradient of a line connecting P and T0.368p. This parameter characteristic of each sample's texture was calculated as the ratio of 'T/P' where, T is the relaxation time and is defined as the time required for a stress at constant strain to decrease to 1/e of its original value, where 'e' is the base of natural logarithms (2.7183). Since 1/e=0.368, the relaxation time is the time required for the force to decay to 36.8% of its original value. P is the peak height of the curve (i.e. the force value at the maximum height). This method was adopted from the bakery industry for testing the degree of gluten development in bread dough. The 'T/P' values obtained over the course of ripening for differently treated salted-herring in barrels ranged between 1 and 12. The trends in 'T/P' value, during ripening period for the different samples, appeared to be parallel changes in texture perceived by sensory observation (subjective measurement), although the heterogeneous nature of the samples gave standard deviations, about the replicate sample mean, around 5%. The method appears promising as an objective measure for monitoring this aspect of the textural quality of barrel salted-herring through ripening if reproducibility of test results can be improved by more careful standardization of sample preparation and test protocol.

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糯小麦在食品加工业、淀粉加工业及其它工业上有着重要用途,是近年来许多国家小麦研究的重要课题。国外糯小麦选育尚未突破高产与糯性相结合的难点,国内目前还没有培育出高蛋白强筋型的糯小麦品种,这在一定程度上与缺乏合适的育种方法和高效、实用的糯小麦分子标记辅助育种技术有关。国内外对Wx基因效应的研究主要利用缺体-四体系、重组自交系或近等基因系,还未见有利用遗传背景相同的BC5F2代回交改良群体的报道。 糯性位点近等基因系是小麦淀粉品质育种的重要材料,而我国目前还没有一套中国栽培小麦遗传背景的糯性位点近等基因系。为了选育部分糯小麦、全糯小麦和中国栽培小麦遗传背景的糯性位点近等基因系,我们利用Wx蛋白电泳和高效实用的分子标记技术体系来鉴定糯小麦杂交后代的基因型,结果证明该体系能有效地用于糯小麦的分子标记辅助育种。以中国春糯性位点全套近等基因系为研究材料,对小麦Wx基因的6个STS标记和1个CAPS标记进行了筛选,改良PCR扩增条件以及产物检测方式后,从这些标记中筛选出3个标记,包括鉴定Wx-A1、Wx-D1位点的2个共显性STS标记和Wx-B1位点的1个显性STS标记。利用上述3个分子标记从BC5F2 代回交改良群体中筛选出了8种Wx基因型,经卡方检验,其分离比符合3对基因的分离比例,其中基因型为aabbdd的植株有2株,直链淀粉含量分别为1.81%和0.82%,为全糯小麦;基因型为AAbbdd,aabbDD的部分糯性植株各有1株,直链淀粉含量分别为15.24%和17.57%。以上4株植株的农艺性状和品质性状接近回交亲本“川育12”,并明显优于全糯材料“98Y1441”,表明采用回交法与Wx基因分子标记辅助选择相结合,有助于培育高产、优质的全糯和部分糯小麦。同时,本研究中建立的分子标记技术体系,也为选育具有中国栽培小麦遗传背景的糯性位点近等基因系奠定了基础。 在基因型鉴定的基础上,利用糯小麦杂交后代BC5F2代回交改良群体研究了各基因缺失降低直链淀粉含量的效果和各基因合成直链淀粉的能力,以及直链淀粉含量与农艺性状、品质性状、淀粉糊化特性等的相关性。缺失不同Wx基因的8种基因型,直链淀粉含量差异显著。研究单缺失基因型发现,减少效应最大的是Wx-B1b基因,Wx-B1b和Wx-D1b基因没有显著差异,减少效应最小的是Wx-A1b基因。研究双缺失基因型发现,Wx基因合成直链淀粉的能力,Wx-B1a基因最高,Wx-A1a基因最低,而Wx-B1a和Wx-D1a基因差异很小;直链淀粉含量与株高、穗长、小穗数、穗粒数、千粒重等农艺性状相关不显著,表明淀粉品质育种可以与高产育种实现有机结合;直链淀粉含量与SDS-沉降值呈显著负相关(r=-0.726),说明直链淀粉含量降低在一定程度上有利于提高小麦营养与加工品质,这一结果至今未见有文献报道;全糯类型的淀粉糊化特性与其他类型显著不同,具有最高的峰值粘度和稀懈值,最低的低谷粘度、最终粘度、反弹值、峰值时间、糊化温度、起始糊化温度,表明糯小麦淀粉在食品和工业上具有特殊用途;稀懈值与直链淀粉含量呈极显著负相关(r=-0.969),其他粘度参数与直链淀粉含量呈显著正相关(最终粘度r=0.797,低谷粘度r=0.910、反弹值r=0.954、峰值时间r=0.970、糊化温度r=0.962、起始糊化温度r=0.932)。以上结论可为糯小麦品种选育和淀粉品质改良提供理论依据。 Waxy wheat is very important in food processing industry, starch processing industry and the other industries, so it is a focus of wheat research in many countries these years. Foreign wheat breeders have not conquered the difficulty of high yield combined with waxy character, and there is no waxy wheat variety with high protein and strong gluten in China at present,all of which were caused by lacking proper breeding methods and effective, applied molecular markers-assisted selection technique at a certain extent. Until now, research about the effect of waxy genes mainly depended on nullisomic-tetrasomic lines, recombinant lines or near-isogenic lines, and it is lacking the reports of using improved BC5F2 backcross progenies under the common genetic background. Near-isogenic lines at the Wx loci are important materials for wheat starch quality breeding. However, there are no such lines under the Chinese cultivated wheat genetic background. To develop partial waxy wheats, waxy wheats and near-isogenic lines at the Wx loci under the Chinese cultivated wheat genetic background, we use SDS-PAGE of waxy proteins and effective, applied molecular marker-assisted selection technical system to identify the genotype of waxy wheat’s progenies. The results indicated that such a system is applicable in waxy wheat’s molecular marker-assisted selection effectively. A series of Chinese Spring Wx loci near-isogenic lines were used to identify the specific bands of six STS markers and one CAPS marker of Wx genes. After optimizing PCR amplification and separating of PCR products, three co-dominant and dominant STS-markers were identified at the Wx-A1, Wx-D1 and Wx-B1 loci, respectively, which were used to identify the genotype of waxy wheat’s progenies. Eight Wx genotypes were developed from the improved BC5F2 backcross progenies, which follows Mendelian segregation. Among them, there were two aabbdd waxy plants whose amylose content in the BC5F3 seeds were 1.81% and 0.82%, respectively. In addition, there were partial waxy plants (AAbbdd and aabbDD) whose amylose content in the BC5F3 seeds were 15.24 % and 17.57%, respectively. Most agronomic and quality traits of these four plants resembled those of the recurrent parent “Chuanyu 12”, and superior to waxy wheat parent “98Y1441”. This shows that backcross approach in combination with molecular marker-assisted selection of waxy genes is helpful to develop partial and full waxy wheat with good traits in the waxy wheat breeding program. At the same time, molecular marker-assisted selection technical system in this paper, also establish the base for breeding the near-isogenic lines at the Wx loci under the Chinese cultivated wheat genetic background. According to the results of genotype identification, we use waxy wheat’s improved BC5F2 backcross progenies to verify the effects of null alleles on reducing amylose content and determine the amylose synthesis capacity of each Wx gene independently, and investigate the relativity among amylose content with agonomic traits, quality traits, starch pasting properties respectively. There was significant difference in amylose content of the eight genotypes carrying different null Wx alleles. The reducing effect of the single null alleles was the most significant in Wx-B1b, followed by Wx-D1b and Wx-A1b, and there was no significant difference between Wx-B1b and Wx-D1b. The results of the double null lines further demonstrated that for the capacity of amylose synthesis, Wx-B1a was the highest, followed by Wx-D1a and Wx-A1a, and there was no significant difference between Wx-B1a and Wx-D1a. Amylose contents of the eight genotypes were not significantly correlated with plant height, spike length, spikelets per spike, grains per spike, 1000-grain weight, which showed that starch quality breeding could integrate with high yield breeding. Amylose contents of the eight genotypes were negatively significantly correlated with SDS-sedimentation value(r=-0.726), which suggested that reduction in amylose content is propitious to improve quality at a certain extent. Starch pasting properties of the full waxy type was significantly different from the other seven types, with the highest peak viscosity and breakdown, and lowest valley viscosity, final viscosity, setback, peak time, pasting temperature and starting pasting temperature. It indicated that waxy wheat starch has special use in food and industry. Breakdown was negatively significantly correlated with amylose contents (r=-0.969), and the other parameters were positively significantly correlated with amylose contents (r=0.797 for final viscosity, r=0.910 for trough viscosity, r =0.954 for setback, r =0.970 for peak time, r=0.962 for pasting temperature and r=0.932 for starting pasting temperature ). The results of the study are very useful for waxy wheat variety breeding and starch quality improvement.

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小麦加工品质改良已成为我国小麦育种的主要目标之一。特别是我国加入WTO以后,对小麦产品的质量提出了更高的要求,小麦品质改良的任务将更加艰巨和重要,小麦胚乳蛋白是影响小麦加工品质性状的重要因素。因此,深入了解小麦胚乳蛋白对加工品质性状的影响及其分子基础,为品质改良提供理论依据和科学指导,对加速我国小麦品质育种和优质小麦生产具有重要意义。本研究选用在麦谷蛋白5个基因位点(Glu-A1、Glu-B1、Glu-D1、Glu-B3和Glu-D3)上均含不同等位基因的小麦品种99G45和京771及Pm97034和京771杂交F9代共164个麦谷蛋白纯合系,及228个中国推广普通小麦品种和高代育成品系为试材,研究了麦谷蛋白Glu-1和Glu-3位点基因等位变异对籽粒蛋白、湿面筋含量、Zeleny沉降值和SDS沉降值间的关系;本研究还利用小麦A、B和D基因组中低分子量麦谷蛋白亚基(LMW-GS)基因特异引物,通过PCR方法克隆了1个Glu-A3位点和3个Glu-B3位点LMW-GS基因片段,在此基础上分析了不同等位基因对品质造成差异的分子基础;另外,本研究对中国近年推广的部分品种和育成的高代品系资源的多样性进行了分析。现将主要研究结果简述如下: 1. 对来自三个麦区的148份材料的醇溶蛋白组成进行了分析,结果表明,各麦区醇溶蛋白模式具有较大差异。在ω区,A7、B、E、F、G、J、P、Q、S和U仅存在于西南秋播麦区;A3、M、N、R、W和X仅存在于黄淮特种麦区;K仅存在于北方冬麦区;A6是北方冬麦区出现频率最高的带型模式,而西南秋播麦区中D出现的频率最高。ω-区的E、H和M几种模式是以前国内外未曾报道的。且初步确定,这些模式对品质性状具有正效应。至于γ区,A、B、D、E和F在各区均有出现,其中B和E在各区出现的频率都很高,在26.1-39.6%之间。相反,H 仅出现在黄淮特种麦区,J仅限于西南秋播麦区。对于β-区醇溶蛋白,B型模式在所有区中都相当高,而模式A仅存在于第三区.对于α-区,模式A在Ⅲ区而模式D在Ⅱ区出现的频率很高。1BL.1RS易位系在中国小麦品种中出现频率高达41.2%,在I, II和Ⅲ麦区的出现频率分别为 45.5、43.5和35.2%。各生态区模式的差异可能是品种适应不同生态条件和人为选择的结果,但这有待进一步证明。由于醇溶蛋白位点(Gli-1)与LMW-GS位点(Glu-3)紧密连锁,本结果可为下面确定普通小麦LMW-GS等位基因变异所用。 2. 利用Gli-1与Glu-3的紧密连锁,以228个小麦品种/系为材料,首次对中国小麦品种麦谷蛋白亚基的6个位点进行综合分析,研究小麦籽粒蛋白与品质性状间的关系,结果表明6个高分子量(HMW)和低分子量(LMW)麦谷蛋白位点对蛋白质含量的效应大小为,Glu-D1>Glu-B3>Glu-A1=Glu-B1> Glu-A3=Glu-D3;对GMP含量的效应大小为, Glu-A3>Glu-B3>Glu-D1> Glu-B1>Glu-A1>Glu-D3;对湿面筋含量的效应大小为, Glu-B1>Glu-B3= Glu-D3>Glu-A3>Glu-A1>Glu-D1;对Zeleny沉降值的效应大小为, Glu-A1> Glu-B3>Glu-D3>Glu-D1>Glu-B1>Glu-A3;对SDS沉降值的效应大小为, Glu-B3>Glu-A1=Glu-D1=Glu-A3>Glu-D3>Glu-B1。对蛋白含量而言,各位点的最佳组合方式为1、17+18、5+10、Glu-A3e、Glu-B3g、Glu-D3b;对湿面筋含量而言,各位点的最佳组合方式为1、6+8、5+10、Glu-A3d、Glu-B3c、Glu-D3b;对Zeleny沉降值而言,各位点的最佳组合方式为N、17+18、5+10、Glu-A3d、Glu-B3d、Glu-D3b;对SDS沉降值而言,各位点的最佳组合方式为1、7+8、2.2+12、Glu-A3b、Glu-B3g、Glu-D3b。另外,分析了稀有亚基对5+12与2.2+12与品质性状的关系,认为5+12对品质有负效应,2.2+12对品质有正效应。在品质育种时,应对优异组合或优异亚基加以利用。 3. 首次利用重组自交系(RILs)为材料,研究麦谷蛋白亚基表达量与品质性状的关系,通过对重组自交系中各HMW-GS表达量的分析,认为,就单个亚基的表达量而言,7亚基最高;其次为2亚基、5亚基、12亚基和10亚基;亚基9和1的表达量最小;N亚基不表达。对成对出现的亚基对而言,x型和y型亚基的总表达量2+12>5+10>7+9>17+18。就单个亚基与品质性状的关系而言,仅有10亚基的表达量与蛋白含量的相关性达5%的显著水平,2亚基的表达量与湿面筋含量呈负相关,显著水平也达5%,其余单个亚基对品质性状均无显著影响;就x型/y型亚基的比例来看,2/12和5/10对湿面筋含量都有显著的负效应;对某一位点等位基因控制的亚基表达总量来看,2+12对SDS沉降值有显著负效应。另外,本研究得出:2+12的亚基对的负效应主要体现在2亚基上,且在同一位点上,x型亚基的表达量大于y型。所以推导稀有亚基组合2+10很可能也是劣质亚基。 4. 以 Glu-A1、Glu-B1、Glu-D1、Glu-B3和Glu-D3作为5个因素对99G45/京771和Pm97034/京771杂交后代的蛋白质含量和SDS沉降值进行多因素方差分析。结果表明,Glu-A1和Glu-D3对蛋白含量的加性效应达5%显著水平;Glu-D1 * Glu-D3对蛋白质含量的互作效应也达5%显著水平;其余位点的加性和互作效应对蛋白质含量的影响均不显著。对SDS 沉降值而言,Glu-D1的加性效应最大,贡献率为4.2 % ,达1 %显著水平,其次是Glu-B1位点,贡献率为3.3% ,达5%显著水平。其余位点对SDS 沉降值的加性和互作效应均未达5%显著水平。总体而言, 各位点对蛋白含量的效应大小为Glu-D3 > Glu-A1 > Glu-D1>Glu-B1>Glu-B3;对SDS沉降值的效应大小为Glu-D1>Glu-B1> Glu-D3>Glu-A1> Glu-B3。Glu-D1和Glu-D3位点上等位基因变异对蛋白含量有显著或极显著影响,含Glu-D1d和Glu-D3 GD、Glu-D3 JD基因的株系分别比含Glu-D1a和Glu-D3 PD基因的株系有较高的蛋白含量;在该遗传背景下,麦谷蛋白各基因位点对蛋白含量的效应大小依次排列为:Glu-A1位点1>N;Glu-B1位点7+9>17+18>14+15;Glu-D1位点5+10>2+12;Glu-B3位点GB>JB>PB;Glu-D3位点GB>JB>PB。对SDS沉降值的效应大小依次排列为:Glu-A1位点1>N;Glu-B1位点7+9=17+18>14+15;Glu-D1位点5+10>2+12;Glu-B3位点GB>JB>PB;Glu-D3位点GB>JB>PB。所以,对蛋白含量和SDS沉降值均较好的组合为1,7+9,5+10,GB,GD。 5. 因为GB和PB对品质的效应有显著差异,选取LMW-GS位点特异扩增引物对京771、99G45和Pm97034的Glu-B3位点进行扩增,结果得到三个不一样的扩增片段(Genebank号为DQ539657-DQ539659),得到的基因片段与Genebank中已报道的同类序列高度同源。通过克隆片段组成的分析,发现对Pm97034的序列较京771和99G45段少一个7氨基酸的重复单元,这可能是它较另外两个片段对面筋强度影响小的主要原因;另外,在99G45的序列中,124位处出现L(亮氨酸)代替P(脯氨酸),158位处出现了T(苏氨酸)代换M(蛋氨酸),这可能是99G45Glu-B3位点序列对SDS沉降值的效应显著优于Pm97034的原因。 6.通过对RILs各位点同普通小麦品种(系)各位点与品质关系的比较,发现对SDS沉降值的效应,各位点在不同研究材料中是不同的,普通小麦中:Glu-B3>Glu-A1=Glu-D1=Glu-A3>Glu-D3>Glu-B1,RILs中:Glu-D1>Glu-B1> Glu-D3>Glu-A1> Glu-B3。利用重组自交系材料(完全排除了1BL/1RS易位干扰)所得到的结果与Gupta and MacRitchie (1994)所得结论一致。进一步证实了1BL/1RS易位对小麦品质的重要影响。对蛋白含量而言,普通小麦品种(系)中,Glu-D1>Glu-B3>Glu-A1=Glu-B1> Glu-A3=Glu-D3,RILs中,Glu-D3 > Glu-A1 > Glu-D1>Glu-B1>Glu-B3,和对SDS沉降值的效应一样,推断在非1BL/1RS易位的情况下,各位点对其效应应为Glu-D3 > Glu-A1 > Glu-D1>Glu-B1>Glu-B3。 对同一位点的等位基因而言,普通小麦和重组自交系中Glu-A1和Glu-D1上的等位基因对品质性状的贡献是一致的,但Glu-B1上的等位基因对SDS沉降值的贡献发生了变化,普通小麦中17+18>7+9,RILs中7+9>17+18,这可能也是1BL/1RS造成的。 Baking quality improved is one of the main object of wheat bread in China. The overall objective of the present studies was to increase the understanding about protein quality in wheat, i.e. to make it possible to improve the production of wheat with desired quality for different end-uses. With the analysis of gluten protein in RILs, 99G45/Jing 771 and Pm97034/Jing, and 228 wheat cultivars or lines in China, the correlations between glutenin compositions and protein content, glutenin macropolymer(GMP), wet gluten content, Zeleny sedimentation value and SDS sedimentation value contentand breadmaking quality were studied. Also a rapid and efficient detection method of geneticpolymorphism at Glu-B3 loci in wheat was established using polymerase chain reaction(PCR).The results obtained were as follows: 1. Cultivated Chinese wheat germplasm has been a valuable genetic resource in international plant breeding. Patterns of gliadin among cultivated Chinese accessions are unknown, despite the proven value and potential novelty. The objective of this work was to analyse the diversity within improved Chinese wheat germplasm. The electrophoretic banding patterns of gliadin in common wheat cultivars and advanced lines were determined by acid-polyacrylamide gel electrophoresis. For 148 leading commercial cultivars and promising advanced lines used in our study, 48 patterns were identified, 29 corresponding to ω-gliadin, 9 to γ-gliadin, 5 to β-gliadin and 5 to α-gliadin. The most frequent patterns were A6 in ω; B in γ; B in β and A in the region of α. 116 band types appeared in the148 samples: 94 accessions had unique gliadin types, and 22 gliadin types while not unique were found in 54 accessions. The gliadin patterns of Chinese wheat cultivars and lines greatly differed from the patterns of wheat lines from other countries. Three patterns, E, J, H, M, N and O in the ω-zone had not previously been reported. Three wheat zones,the Northern Winter Wheat Region, the Yellow and Huai Valley River valleys Winter Wheat Region and the Southwestern Winter Wheat Region,in China showed different frequencies in their gliadin patterns. This information can be used to monitor genetic diversity with Chinese wheat germplasm. 2. To analyse the relationship between the loci and characteristics quality, we utilized the 228 cultivars/lines. The results showed that : For protein content, Glu-D1 >Glu-B3>Glu-A1=Glu-B1>Glu-A3=Glu-D3. For GMP content, Glu-A3>Glu-B3 >Glu-D1>Glu-B1>Glu-A1>Glu-D3. For wet gluten content, Glu-B1>Glu-B3= Glu-D3>Glu-A3>Glu-A1>Glu-D1. For Zeleny sedimentation value, Glu-A1>Glu-B3> Glu-D3>Glu-D1>Glu-B1>Glu-A3, For SDS sedimentation value, Glu-B3>Glu-A1= Glu-D1= lu-A3>Glu-D3>Glu-B1。For protein content, the best combination of 6 loci is (1,17+18,5+10,Glu-A3e, Glu-B3g,Glu-D3b). For wet gluten content, the best combination of 6 loci is (1,6+8,5+10,Glu-A3d,Glu-B3c,Glu-D3b). For Zeleny sedimentation value, the best combination of 6 loci is (N,17+18,5+10,Glu-A3d, Glu-B3d, Glu-D3b). For SDS sedimentation value, the best combination of 6 loci is(7+8,2.2+12,Glu-A3b, Glu-B3g,Glu-D3b)。Additional, we analysed the relationship between the subunits 5+12 and 2.2+12, think that 5+12 was negative for quality, 2.2+12 is postive for quality. It should be effective utilized. 3. It’s the first time to utilize RILs to study the relationship between subunits expression quantity and characteristics quality. The results showed that: For single subunit, the expression quantity of 7 is the highest. Then the 2, 5, 12 and 10. The expression of subunit 9 and 1 is the lowest. Subunit N is not expressed. For subunits, the expression quantity of x type and y type are 2+12>5+10>7+9>17+18. The significant relation of 5% only showed between the expression quantity of subunit 10 and protein content. The relationship between expression quantity of others and characteristic quality was not significant. For x type/ytype, 2/12 and 5/10 is negative relation insignificant level. For the subunit(s) in a loci, Only 2+12 effect SDS sedimentation value negative in significant level. 4. With RILs 99G45/Jing 771 and Pm97034/Jing 771, we found that: The effective of Glu-A1, Glu-D3 and Glu-D1 * Glu-D3 for protein content is significant at 5% level. The effect of other loci for protein wre not significant. For SDS sedimentation value, the effect of Glu-D1is the highest, which contribution is 4.2 % .Then the Glu-B1, contribution is 3.3%. The effect of other loci for SDS sedimentationvalue were not significant. In total, for protein content: Glu-D3 > Glu-A1 > Glu-D1>Glu-B1>Glu-B3; for SDS sedimentationvalue: Glu-D1>Glu-B1> Glu-D3>Glu-A1>Glu-B3. The effect of alleles in Glu-D1 and Glu-D3 loci are significant at 1% or 5%. In Glu-A1, 1>N; Glu-B1, 7+9>17+18>14+15; Glu-D, 5+10>2+12; Glu-B3, GB>JB>PB; Glu-D3, GB>JB>PB. For SDS sedimentation, Glu-A1, 1>N; Glu-B1, 7+9=17+18>14+15; Glu-D1, 5+10>2+12; Glu-B3, GB>JB>PB; Glu-D3, GB>JB>PB. The best combinations for SDS sedimentation value is 1,7+9,5+10,GB,GD. 5. Because of the difference of GB and PB for SDS sedimentation value, we selected the specific primer for LMW-GS loci to amplified the Glu-B3 of Jing771, 99G45and Pm97034. We got 3 amplify fragment (Gene Bank accession number are DQ539657-DQ539659). We found that the fragment of Pm97034 were deleted a repetitive 7 amino acid domain, which is perhaps the reason effect the gluten strength. Furthermore, in the position 124 of sequence 99G45, L has been replaced with P. Position 158, T replaced M, which may be the reason why the Glu-B3 locus of 99G45 is prefer to Pm97034 when refer to SDS sedimentation value. 6. Comparing the results of RILs and common wheat, we found that perhaps just the1BL/1RS made the difference of loci in different accession.

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A cultivar de trigo BRS 49, desenvolvida pela Embrapa Trigo, foi indicada para semeadura no Estado do Parana, a partir de 1999. Para a determinacao do seu valor de cultivo e uso, foram utilizados dados de 54 experimentos, instalados em nove locais das regioes 6, 7 e 8 do Parana, no periodo de 1996 a 1998. Apresenta como principais caracteristicas: ciclo intermediario, altura media a alta, moderada resistencia ao acamamento, tolerancia ao aluminio toxico no solo, moderada resistencia ao oidio (Blumeria graminis f. sp tritici) e a ferrugem da folha (Puccinia recondita f. sp. tritici), com percentuais de severidade em ensaios inferiores aos das testemunhas Trigo BR 23 e Trigo BR 35, suscetibilidade a giberela (Giberella zeae), cujas notas foram superiores as atribuidas as testemunhas, elevado potencial de rendimento e boa qualidade industrial. No trienio considerado, apresentou rendimento medio de 4.667 kg/ha na Regiao 6, 3.432 kg/ha, na Regiao 7, e 3.930 kg/ha, na regiao 8. Esses valores foram 11%, 11% e 28%, respectivamente, superiores a media das testemunhas (Trigo BR 2, Trigo BR 35 e CEP 24-Industrial). Os percentuais de resposta ao controle fitossanitario foram, em geral, inferiores aos observados nas referidas testemunhas, evidenciando o elevado potencial de rendimento da cultivar com aplicacoes reduzidas de fungicidas. A forca geral de gluten (W), na media de 14 amostras coletadas nas tres Regioes, foi de 260 e a relacao P/L foi de 0,685. Portanto, a cultivar se enquadra na classe Trigo Pao, de acordo com a Instrucao Normativa n.1, de 27 de janeiro de 1999 do MA. A cultivar apresenta caracteristicas de interesse agronomico, principalmente por possibilitar a obtencao de altos rendimentos, sem a necessidade de elevados investimentos com fungicidas.

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Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

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The use of unmalted oats or sorghum in brewing has great potential for creating new beer types/flavors and saving costs. However, the substitution of barley malt with oat or sorghum adjunct is not only innovative but also challenging due to their specific grain characteristics. The overall objectives of this Ph.D. project were: 1) to investigate the impact of various types and levels of oats or sorghum on the quality/processability of mashes, worts, and beers; 2) to provide solutions as regards the application of industrial enzymes to overcome potential brewing problems. For these purposes, a highly precise rheological method using a controlled stress rheometer was developed and successfully applied as a tool for optimizing enzyme additions and process parameters. Further, eight different oat cultivars were compared in terms of their suitability as brewing adjuncts and two very promising types identified. In another study, the limitations of barley malt enzymes and the benefits of the application of industrial enzymes in high-gravity brewing with oats were determined. It is recommended to add enzymes to high-gravity mashes when substituting 30% or more barley malt with oats in order to prevent filtration and fermentation problems. Pilot-scale brewing trials using 10–40% unmalted oats revealed that the sensory quality of oat beers improved with increasing adjunct level. In addition, commercially available oat and sorghum flours were implemented into brewing. The use of up to 70% oat flour and 50% sorghum flour, respectively, is not only technically feasible but also economically beneficial. In a further study on sorghum was demonstrated that the optimization of industrial mashing enzymes has great potential for reducing beer production costs. A comparison of the brewing performance of red Italian and white Nigerian sorghum clearly showed that European grown sorghum is suitable for brewing purposes; 40% red sorghum beers were even found to be very low in gluten.

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Quinoa (Chenopodium quinoa) is a seed crop native to the Andes, that can be used in a variety of food product in a similar manner to cereals. Unlike most plants, quinoa contains protein with a balanced amino acid profile. This makes it an interesting raw material for e.g. dairy product substitutes, a growing market in Europe and U.S. Quinoa can however have unpleasant off-flavours when processed into formulated products. One means of improving the palatability is seed germination. Also, the increased activities of hydrolytic enzymes can have a beneficial influence in food processing. In this thesis, the germination pattern of quinoa was studied, and the influence of quinoa malt was evaluated in a model product. Additionally, to explore its potential for dairy-type products, quinoa protein was isolated from an embryo-enriched milling fraction of non-germinated quinoa and tested for functional and gelation properties. Quinoa seeds imbibed water very rapidly, and most seeds showed radicle protrusion after 8-9 h. The α-amylase activity was very low, and started to increase only after 24 hours of germination in the starchy perisperm. Proteolytic activity was very high in dry ungerminated seeds, and increased slightly over 24 h. A significant fraction of this activity was located in the micropylar endosperm. The incorporation of germinated quinoa in gluten-free bread had no significant effect on the baking properties due to low α-amylase activity. Upon acidification with glucono-δ-lactone, quinoa milk formed a structured gel. The gelation behaviour was further studied using a quinoa protein isolate (QPI) extracted from an embryoenriched milling fraction. QPI required a heat-denaturation step to form gel structures. The heating pH influenced the properties drastically: heating at pH 10.5 led to a dramatic increase in solubility, emulsifying properties, and a formation of a fine-structured gel with a high storage modulus (G') when acidified. Heating at pH 8.5 varied very little from the unheated protein in terms of functional properties, and only formed a randomly aggregated coagulum with a low G'. Further study of changes over the course of heating showed that the mechanism of heat-denaturation and aggregation indeed varied largely depending on pH. The large difference in gelation behaviour may be related to the nature of aggregates formed during heating. To conclude, germination for increased enzyme activities may not be feasible, but the structure-forming properties of quinoa protein could possibly be exploited in dairy-type products.

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BACKGROUND: Tissue transglutaminase (t-TG) is the main autoantigen recognized by the endomysium antibodies (EMA) observed in patients with celiac disease (CD). The aim of the study was to assess an ELISA method for t-TG antibodies (t-TGA) with respect to EMA IF assay in pediatric and adult patients. METHODS: t-TGA were analyzed by ELISA in 220 sera samples: 82 patients with biopsy-proven untreated CD (23 adults and 59 children), 14 CD children on gluten-free diet, 18 asymptomatic relatives of CD patients, and 106 age-matched control patients with gluten-unrelated gastrointestinal diseases (58 adults and 48 children). Serum IgA EMA were tested on umbilical cord sections in all patients. RESULTS: The great majority (92.7%) of untreated CD patients (both adults and children) were t-TGA positive (values ranging from 20.1 to > 300 AU). None of the child control patients and only two out of 58 (3.4%) of the adults with unrelated gastrointestinal diseases had serum t-TGA positivity; two out of 18 first-degree relatives with biopsy-proved silent CD were t-TGA (as well as EMA) positive. Finally, two out of 14 CD children, assuming a gluten-free diet, had serum t-TGA (as well as EMA). A highly significant correlation (P < 0.001) was observed between t-TGA concentrations and EMA. t-TGA showed a sensitivity of 87% and 95%, a specificity of 97% and 100% for adults and children, respectively. CONCLUSION: The method is highly sensitive and specific in the diagnosis of CD and is promising as a tool for routine diagnostic use and population screening, especially in children.

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El genotipo (G), el ambiente (A) y la interacción G x A pueden influir de manera diferente sobre las característica que definen la calidad comercial e industrial de trigo pan. Los objetivos de esta tesis fueron: 1) Caracterizar el rendimiento y a sus componentes en cultivares de trigo pertenecientes a diferentes grupos de calidad, expuestos a ofertas de nitrógeno contrastantes. 2) Estudiar el impacto de distinta disponibilidad de nitrógeno sobre los componentes fisiológicos del llenado de los granos (i.e tasa y duración) en distintaas variedades de trigo pan y su posible efecto sobre los parámetros de calidad. 3) Caracterizar y cuantificar la interacción genotipo por ambiente sobre la expresión de los parámetros que determinan el rendimiento y la calidad comercial e industrial del trigo pan en ambientes con diferentes disponibilidades de nitrógeno. Se realizaron ensayos en dos localidades, durante dos años, utilizando seis variedades de distinta aptitud panadera (2 de cada grupo de clasificación por grupo de calidad -GC-), aplicando cuatro tratamientos de fertilización nitrogenadas. Se evaluó el efecto genético, ambiental y la interacción G x A, sobre el rendimiento y sus componentes, el peso de los granos y sus componentes y sobre los parámetros de calidad comercial e industrial. Los resultados mostraron que el rendimiento y sus componentes (número de granos, biomasa aérea, eficiencia de uso de la radiación interceptada acumulada) fueron afectados principalmente por el ambiente y el manejo nutricional dentro de de cada ambiente. Para el peso de los granos y sus componentes (tasa y duración) el efecto del manejo del nitrógeno no fue importante, aunque sí lo fue el efecto genotipo. Para los parámetros de calidad el efecto genotipo fue más importante solo para la tenacidad, mientras que el peso hectolítrico, gluten húmedo, fuerza panadera, la relación de equilibrio (P/L) y volumen de pan fueron modificados principalmente por el efecto ambiente no manejable como son el año y la localidad, en tanto la proteína fue afectada principalmente por el factor ambiental asociado al manejo nutricional. La interacción GxA fue el efecto que explicó en mayor medida las variaciones de rendimiento de harina, absorción de agua y tiempo de amasado. La fuerte interacción GxA observada para la mayoría de los parámetros de calidad determinó que variedades de un determinado GC cambien de grupo asociado principalmente a factores ambientales como la localidad y el año, mientras que el manejo nutricional tuvo un impacto menor