Master of Science / Department of Plant Pathology / William Bockus / Barley yellow dwarf (BYD) is one of the most important wheat diseases in the state of Kansas. Despite the development of cultivars with improved levels of resistance to BYD, little is known about the impact that this resistance has on yield loss from the disease. The intent of this research was to estimate yield loss in winter wheat cultivars in Kansas due to BYD and quantify the reduction in losses associated with resistant cultivars. During seven years, BYD disease incidence was visually assessed on numerous winter wheat cultivars in replicated field nurseries. Cultivars were planted about three weeks early to promote disease. When grain yields were regressed against BYD incidence scores, negative linear relationships significantly fit the data for each year and for the combined dataset covering all seven years. The models showed that, depending upon the year, 19-48% (average 33%) of the yields was explained by BYD incidence. For the combined dataset, 29% of the relative yield was explained by BYD incidence. The models predicted that cultivars showing high disease incidence had 25-86% (average 49%) less yield than a hypothetical cultivar that showed zero incidence. Using the models, the moderate level of resistance in the cultivar Everest was calculated to reduce yield loss from BYD by about 73%. Therefore, utilizing visual BYD symptom evaluations in Kansas, coupled with grain yields, is useful to estimate yield loss from the disease. Furthermore, linear models that incorporate those parameters can be used to calculate the impact of improving cultivar resistance to BYD on yield losses.

O teste de exercício cardiopulmonar (TECP) (ou teste ergoespirométrico) é considerado o padrão-ouro para avaliação da potência aeróbia, contudo este teste demanda equipamentos sofisticados e profissionais especializados. O interesse pelo teste do degrau para avaliação da capacidade física em indivíduos com doenças pulmonares crônicas cresceu nos últimos anos devido à sua portabilidade. Entretanto, o uso do teste do degrau incremental (TDIM) para avaliação da capacidade física de indivíduos com asma permanece desconhecido. O objetivo deste estudo foi avaliar as respostas cardiopulmonares e metabólicas durante o TDIM em indivíduos com asma moderada a grave bem como avaliar sua reprodutibilidade nesta população. Cinquenta e quatro indivíduos com asma moderada a grave foram submetidos ao TECP e a dois TDIM em ordem aleatorizada. Os testes foram realizados em 2 dias não consecutivos com intervalo mínimo de 48 horas. No primeiro dia, os indivíduos realizaram avaliação do controle clínico da asma pelo Asthma Control Questionnaire (ACQ) e o teste TECP ou os 2 TDIM, de acordo com a aleatorização. Na segunda visita, os indivíduos foram submetidos aos 2 TDIM ou ao TECP de acordo com a aleatorização. Os dados dos pacientes foram analisados em subgrupos de acordo com o índice de massa corpórea (IMC) e divididos em eutróficos, sobrepeso e obesos. O desempenho no melhor TDIM (m-TDIM) foi comparado à resposta do TECP. Os valores de consumo de oxigênio no pico do exercício (VO2pico) obtidos no m-TDIM foram inferiores ao TECP (2.042±494; 1.749±434 ml/min, média±DP, respectivamente; p=0,01). Os limites de concordância do VO2pico entre o m-TDIM e TECP variaram de -786 a 201 ml/min. Não foi observada diferença nos indivíduos eutróficos com asma (EA) na maioria das variáveis analisadas, exceto no coeficiente respiratório (RER). Por outro lado, foi verificado que a maioria das variáveis obtidas no TDIM foram inferiores nos indivíduos com sobrepeso e obeso comparado ao TECP. Apesar dos valores inferiores observados no m-TDIM quando comparado ao TECP na análise com todos os indivíduos, o TDIM se mostrou reprodutível e apresentou um excelente coeficiente de correlação intraclasse para o número de degraus (0,88[0,79 - 0,93]), o VO2pico (0,93[0,88 - 0,96]) e a frequência cardíaca máxima (0,86 [0,76 - 0,92]). Os nossos resultados mostram que o TDIM induz a uma menor resposta do VO2 pico quando comparado ao TECP em indivíduos com asma moderada a grave independente do IMC. Contudo, o TDIM foi bem tolerado e reprodutível nesta população / The cardiopulmonary exercise test (CPET) is the gold standard to measure aerobic fitness; however, CPET is expensive and requires specialized equipment and a qualified operator. There is a growing interest in using step tests to assess exercise capacity in subjects with chronic lung diseases due to its portability. However, the use of modified incremental step test (MIST) to evaluate exercise capacity in subjects with asthma remains unknown. This study aims to evaluate cardiopulmonary and metabolic responses during MIST in adults with moderate to severe asthma. In addition, we investigated the reliability of the MIST in this population. Fifty-four adults with asthma performed a CPET and two MIST with pulmonary gas analysis on two separate days in a randomized order. The tests were performed in 2 non-consecutive days with a minimum interval of 48 hours. On the first day was evaluated the clinical control of asthma by the Asthma Control Questionnaire (ACQ) and the TECP or 2 TDIM, according to the randomization. At the second visit, the subjects were submitted to 2 TDIM or TECP according to the randomization. The subjects were allocated according to BMI (body mass index) in eutrophic, overweight and obese. The best MIST (b-MIST) was considered at the peak of the exercise for comparison with CPET. There was a difference in peak oxygen uptake (VO2peak) assessed by the CPET and the b-MIST (2,042±494; 1,749±434 ml/min, mean ± SD, respectively; p=0.01). The limits of agreement for VO2peak between the CEPT and the b-MIST with ranged from -786 to 201ml/min. MIST was reproducible and presented an excellent intraclass correlation coefficient (CCI [95% confidence interval]) for the number of steps (0.88[0.79 - 0.93]), VO2 (0.93[0.88 - 0.96]) and maximal heart rate (0.86 [0.76 - 0.92]). The MIST elicits a lower VO2peak than the CPET in subjects with moderate to severe asthma regardless the BMI. However, the MIST is well tolerated and reproducible in this population

Neste trabalho foi realizado um estudo sobre a associação entre características da madeira importantes na produção de polpa celulósica e o genótipo dos indivíduos de uma população de híbridos de Eucalyptus grandis × Eucalyptus urophylla aos quatro anos de idade. Em termos de características da madeira determinou-se a densidade básica, teor de lignina klason, extrativos totais e holocelulose. A genotipagem foi realizada utilizando 14 combinações entre o iniciador arbitrário TRAP2 e iniciadores fixos relacionados às características de interesse. 36 marcadores microssatélites também foram utilizados na genotipagem. A densidade básica bem como os teores dos componentes químicos da madeira encontrados estão dentro do esperado para espécies do gênero. O estudo de associação fenótipo-genótipo, detectou oito associações, sendo quatro delas entre marcadores TRAP e as quatro entre marcadores microssatélites. As associações significativas detectadas foram principalmente entre marcadores e densidade básica sendo as maiores associações com os marcadores TRAP2/COMT (210 pb) (26%) e E2010 (24%). Igualmente e, em segundo lugar, entre marcadores e teor de lignina e extrativos totais. O maior grau de associação significativo foi detectado entre o marcador TRAP2/HCT (190 pb) e o teor de lignina sendo a associação de 32 %. Não foi detectada associação entre marcadores e teor de holocelulose. Mais de uma marca foi relacionada com uma característica o que reforça teorias de controle genético exercido por mais de um gene. Também foi detectada associação entre uma marca e duas características diferentes sugerindo que um mesmo gene possa exercer o controle sobre mais de uma característica. O estudo mostrou portanto correlações entre as propriedades da madeira e associação, em diferentes níveis, entre essas e marcas genéticas. / It was studied in this work the association between wood characteristics that are important to the cellulosic pulp and paper production and the hybrid population individual genotype Eucalyptus grandis × Eucalyptus urophylla at the age of four years. The basic density, characteristic of mayor interest, was valuated using the maximum humidity content method. The amount of Klason lignin, total extract and holocellulose were obtained using conventional analysis methods. The genotyping was done using 14 combinations between the arbitrary primer TRAP2 and fixed primers related to the characteristics of interest. 36 microsatellite markers were also used during the genotyping process. The basic density and the wood chemical components amounts were found between the expected values for each genus. The study revealed that there are negative co-relationships between the wood chemical characteristics. The relationship between the holocellulose amount and the total extracts value is approximately 71% as the corelationship between the holocellulose amount and the lignin value is 55%. It was detected a positive relationship between the Klason lignin amount and the basic density. The phenotypegenotype association study detected eight associations, four of them being between TRAP markers and the other four between microsatellite markers. The significant associations detected were mainly between the markers and the basic density, especially the association with the markers TRAP2/COMT (210 bp) (26%) and E2010 (24%). Equally and in second place it stands the association between the markers and the lignin value and the total extracts. The mayor significant association degree was detected between the TRAP2/HCT marker (190 bp) and the lignin value, being it 32 %. It was not detected any relationship between the markers and the holocellulose amount. More than one mark was related to one characteristic which enhances genetic control theories carried by more than one gene. It was also detected an association between one mark and two different characteristics suggesting that one same gene can have control over more than one characteristic. The study revealed therefore co-relationships between wood properties and association in different levels between those and genetic marks.

A perda auditiva é uma característica importante da Síndrome de Stickler. Na literatura, poucos trabalhos têm caracterizado essa perda, e não há estudos do efeito de outros fatores de risco sobre a audição no Stickler. Realizou-se este trabalho com a finalidade de verificar o fenótipo clínico e audiológico de um grupo de 26 crianças, com idade média de cinco anos e sete meses, afetadas pela SS, no Hospital de Reabilitação de Anomalias Craniofaciais. Após análise dos dados dos exames clínico e audiológico, concluiu-se que 80,76% (IC 95%, 60, 65 – 93, 44) das crianças apresentaram perda auditiva, sendo 34,61% (IC 95%, 17, 21 – 55, 66), de perda sensorioneural associada à SS, tipicamente para altas freqüências; 11,53% (IC 95%, 2, 44 – 30, 15) de perda mista, e 34,61% (IC 95%, 17, 21 – 55, 66) de perda auditiva condutiva, que é comum nas crianças com SS, devido à presença da fissura de palato e outras anomalias craniofaciais, levando a uma disfunção da orelha média com o comprometimento da sensibilidade auditiva. O grau de perda auditiva variou de leve a grave. A análise estatística pelo teste de Fisher (p<0,05) não revelou qualquer efeito significativo da exposição das crianças a outros fatores de risco de perda auditiva sobre a perda sensorioneural, mostrando ser este tipo de perda auditiva característica da SS. Na impossibilidade da realização do exame genético molecular, o fenótipo clínico, associado à avaliação audiológica, pode sugerir o diagnóstico do tipo da SS. / Hearing loss is an important characteristic of the Stickler’s Syndrome. In literature, few studies have defined the characterization of this loss, and there are no studies about the effect of other risk factors of hearing in Sticker. This present work has been made with the purpose of verifying the clinical and audiological phenotype of a group of 26 children with average age of 5 years and 7 months, affected by SS, at the Hospital de Reabilitação de Anomalias Craniofaciais. After analyzing the data of the clinical and audiological exams, we concluded that 80.76% (IC 95%, 60,65 – 93,44) of the children have presented hearing loss, being 34.61% (IC 95%, 17,21 – 55,66) of sensorial neural loss, associated with SS, typically for high frequencies; 11.53% (IC 95%, 2,44 – 30,15) of mixed loss; and 34.61% (IC 95%, 17,21 – 55,66) of conductive hearing loss, which is common in children with SS, due to the presence of cleft palate and other craniofacial anomalies, leading to a disfunction of the mid ear, with harming of hearing sensibility. The degree of hearing loss has varied from mild to severe. The statistical analysis through Fisher’s test (p<0.05) didn’t reveal any significant effect of the exposition of children to other risk factors of hearing loss to the sensorial neural loss of SS. Due to the impossibility of the realization of the molecular genetic exam, the clinical phenotype, associated with the audiological evaluation, may suggest the diagnosis of the kind of SS.

A mutagênese química utilizando o agente mutagênico N-ethyl-N-nitrosourea (ENU) seguida da observação do fenótipo deu origem a um mutante Anêmico. O tipo de herança é autossômica dominante, com morte intra útero dos mutantes homozigotos. O mapeamento genético foi feito utilizando-se marcadores microssatélites, sendo selecionados marcadores polimórficos entre as linhagens BALB/c e C57BL/6 envolvidas no mapeamento. Estabeleceu-se um painel de microssatélites distribuídos por todo o genoma do camundongo, que permitisse a localização do cromossomo portador da mutação. O gene mutante foi localizado no cromossomo 7 entre os marcadores D7Mit301 e D7Mit131 delimitando um intervalo entre 46,5cM e 51cM de 4,5cM. Através das análises fenotípicas do mutante Anêmico e estudo dos genes candidatos neste intervalo, foi selecionado o gene Hbb responsável pela síntese das globinas b-major e b-minor , sendo o gene que mais se identifica com as características do mutante, localizado a 50cM. A deficiência deste gene leva a uma das mais severas anemias humana, a b-Talassemia major. / Chemical mutagenesis, using the mutagenic agent N-ethyl-N-nitrosourea (ENU), and followed by observation of the phenotype, originated in an Anaemic mutant. The inheritance-type is dominant auto-somic, with intra-uterus death of the homozygotic mutants. Genetic mapping was undertaken by means of micro-satellite markers, polymorphic markers being selected from among the BALB/c and C57BL/6 lineages involved in the mapping itself. A panel was established of the micro-satellites distributed throughout the whole mouse genome, thereby permitting localization of the mutation bearing chromosome. The mutant gene was located in chromosome 7 between markers D7Mit301 and D7Mit131, these delimiting an interval between 46,5cM and 51cM of 4,5cM. Selection of the Hbb gene responsible for synthesis of the b-major and b-minor globins came about through phenotypic analysis of the Anaemic mutant and a study of candidate genes within this interval, the selected gene being that which was most identified with the mutants characteristics and located at 50cM. A deficiency in this gene leads to one of the most severe forms of human anaemia, b-Talhassemia major.

Introdução: A dermatite atópica (DA) é uma enfermidade cutânea inflamatória de caráter crônico, na qual o prurido é constante, e com marcada xerose. Dermatose que geralmente se inicia na infância, e pode surgir em indivíduos com história pessoal ou familiar de asma, rinite alérgica e/ou DA. A pele com DA apresenta colonização por Staphylococcus aureus (S. aureus) em 80-100% dos casos, sendo responsável pela produção enterotoxinas, capazes de exacerbar a resposta inflamatória na DA. Nesta enfermidade, existem distintos subtipos de células apresentadoras de antígeno ou dendríticas (DC), tanto na pele quanto circulantes. As DC exercem papel relevante na inflamação da DA, em especial um subgrupo de células dendríticas mieloides (mDC), as chamadas células dendríticas inflamatórias epidérmicas (IDEC). Objetivo: Avaliar o fenótipo e a função das mDC (IDEC-like) em células mononucleares do sangue periférico (PBMC) na DA do adulto. Métodos: Foram selecionados 21 pacientes com DA (idades entre18 e 65 anos, sendo 13 homens e oito mulheres) e 21 controles (idades entre 21 e 41 anos, sendo oito homens e 13 mulheres), nos quais foram realizadas as avaliações fenotípica e funcional das mDC (IDEC-like) em PBMC. Para tal, foram analisadas as expressões de: Fc?RI, TNF, IFN-y, IL-10, CD36 e CD83 nas mDC, estimuladas com enterotoxina estafilocócica B (SEB), agonistas de TLR2 (Pam3CSK4), TLR4 (LPS) e de TLR7/8 (CL097) através da citometria de fluxo. Resultados: Os principais achados nos pacientes com DA foram: aumento da frequência de células IDEC-like frente ao estímulo com agonista de TLR2 (Pam3CSK4); aumento da frequência de IFN-y em condição não estimulada, e de IL-10 frente a estímulo com agonista de TLR7/8 (CL097) nesta população de células dendríticas. Conclusão: A caracterização das mDC circulantes na DA evidencia perfil pró-inflamatório em condição não estimulada, impactando na resposta imune adaptativa. O aumento significativo na frequência de células IDEC-like nos pacientes com DA sugere sua participação na perpetuação do processo inflamatório da DA / Introduction: Atopic dermatitis (AD) is an inflammatory skin disease with a chronic course, with constant pruritus and marked xerosis. It usually starts during childhood, and a personal or familial history of skin and/or respiratory allergy may be present. Around 80-100% of the patients show a cutaneous colonization of Staphylococcus aureus (S. aureus), which produces enterotoxins that may exacerbate the inflammatory response in AD. In this disease, there are distinct subtypes of antigen-presenting cells or dendrytic cells (DC), either circulating or present in the affected tissue. DC exert a relevant role in AD inflammation, especially a subgroup of myeloid cells (mDC), known as epidermal inflammmatory dendritic cells (IDEC). Objective: To evaluate the phenotype and function of mDC (IDEC-like) in mononuclear cells of the peripheral blood (PBMC) of adults with AD. Methods: Twenty-one adults with AD (age 18/65; male/female: 13/8) and 21 healthy controls (age 21/41; male/female: 8/13) were selected for the current study. Phenotypical and functional analysis of mDC (IDEC-like) of PBMC were performed, through the expression of Fc?RI ,TNF, IFN-y, IL-10, CD36 and CD83 in mDC, stimulated with enterotoxin B (SEB) and with agonists of TLR2 (Pam3CSK4), TLR4 (LPS) and TLR7/8 (CL097) by flow cytometry. Results: Main findings of AD patients included: elevation of IDEC-like cell frequency with TLR2 (Pam3CSK4) agonist, augmented unstimulated frequency of IFN-y, and of IL-10 with TLR7/8(CL097) agonist of this population of dendritic cells. Conclusion: Characterization of circulating mDC on AD shows proinflammatory profile in unstimulated conditions, therefore causing impact on the adaptive immune response. The significant increase in the frequency of IDEC-like cells in AD patients suggest a role in the maintenance of inflammation in AD

A predição da variância fenotípica é de grande importância para que os sistemas de produção de bovinos de corte consigam aumentar a rentabilidade otimizando o uso de recursos. Modelos mecanicistas dinâmicos do crescimento bovino vêm sendo utilizados como ferramentas de suporte à tomada de decisão em sistemas de manejo individual do gado. Entretanto, a aplicação desses modelos ainda fundamenta-se em parâmetros populacionais, sem qualquer abordagem para que se consiga capturar a variabilidade entre sujeitos nas simulações. Assumindo que modelos mecanicistas sejam capazes de simular o componente de desvio ambiental da variância fenotípica e considerando que marcadores SNPs possam predizer o componente genético dessa variância, esse projeto objetivou evoluir em direção a um modelo matemático que considere a variabilidade entre animais em seu nível genético. Seguindo conceitos de fisiologia genômica computacional, nós assumimos que a variância genética da característica complexa (i.e. produto do comportamento do modelo) surge de características componentes (i.e. parâmetros dos modelos) em níveis hierárquicos mais baixos do sistema biológico. Esse estudo considerou dois modelos mecanicistas do crescimento de bovinos - Cornell Cattle Value Discovery System (CVDS) e Davis Growth Model (DGM) - e ao questionar se os parâmetros de tais modelos mapeariam regiões genômicas que englobam QTLs já descritos para a característica complexa, verificou as suas interpretações biológicas esperadas. Tal constatação forneceu uma prova de conceito de que os parâmetros do CVDS e do DGM são de fato fenótipos cuja interpretação pode ser confirmada através das regiões genômicas mapeadas. Um método de predição genômica foi então utilizado para computar os parâmetros do CVDS e do DGM. Os efeitos dos marcadores SNPs foram estimados tanto para os parâmetros quanto para os fenótipos observados. Nós buscamos qual o melhor cenário de predição - simulações dos modelos com parâmetros computados a partir das informações genômicas ou predição genômica conduzida diretamente nos fenótipos complexos. Nós encontramos que enquanto a predição genômica dos fenótipos complexos pode ser uma melhor opção em relação aos modelos de crescimento, simulações conduzidas com parâmetros obtidos a partir de dados genômicos estão condizentes com simulações geradas com parâmetros obtidos a partir de métodos regulares. Esse é o principal argumento para chamar atenção da comunidade científica de que a abordagem apresentada nesse projeto representa um caminho para o desenvolvimento de uma nova geração de modelos nutricionais aplicados capazes de capturar a variabilidade genética entre bovinos de corte confinados e produzir simulações com variáveis de entrada específicas de cada genótipo. Esse projeto é a primeira abordagem no Brasil conhecida dos autores a usar genótipos Bos indicus para o estudo da aplicação de genômica integrada à modelos mecanicistas para o manejo e comercialização de animais na pecuária. / The prediction of phenotypic variance is important for beef cattle operations to increase profitability by optimizing resource use. Dynamic mechanistic models of cattle growth have been used as decision support tools for individual cattle management systems. However, the application of such models is still based on population parameters, with no further approach to capture between-subject variability. By assuming that mechanistic models are able to simulate environmental deviations components of phenotypic variance and considering that SNPs markers may predict the genetic component of this variance, this project aimed at evolving towards a mathematical model that takes between-animal variance to its genetic level. Following the concepts of computational physiological genomics, we assumed that genetic variance of the complex trait (i.e. outcome of model behavior) arises from component traits (i.e. model parameters) in lower hierarchical levels of biological systems. This study considered two mechanistic models of cattle growth - Cornell Cattle Value Discovery System (CVDS) and Davis Growth Model (DGM) - and verified their expected biological interpretation by asking whether model parameters would map genomic regions that harbors QTLs already described for the complex trait. This provided a proof of concept that CVDS and DGM parameters are indeed phenotypes whose expected interpretations may be stated by means of their mapped genomic regions. A method of genomic prediction to compute parameters for CVDS and DGM was then used. SNP marker effects were estimated both for their parameters and observed phenotypes. We looked for the best prediction scenario - model simulation with parameters computed from genomic data or genomic prediction on complex phenotypes directly. We found that while genomic prediction on complex phenotypes may still be a better option than predictions from growth models, simulations conducted with genomically computed parameters are in accordance with those performed with parameters obtained from regular methods. This is the main argument to call attention from the research community that this approach may pave the way for the development of a new generation of applied nutritional models capable of representing genetic variability among beef cattle under feedlot conditions and performing simulation with inputs from individual\'s genotypes. To our knowledge, this project is the first of this kind in Brazil and the first using Bos indicus genotypes to study the application of genomics integrated with mechanistic models for the management and marketing of commercial livestock.

NTRODUÇÃO: Neoplasias mucinosas primárias do ovário apresentam muitos pontos de controvérsias em relação ao seu padrão de diferenciação, sendo classificadas como tumores benignos, borderline e malignos.Elas também são classificadas em diferentes fenótipos, recentemente designadas como gastrointestinal e seromucinoso. Sua heterogeneidade tem produzido não somente dificuldades na classificação morfológica e no diagnóstico diferencial com neoplasias metastáticas, mas também na compreensão da patogênese e na interpretação imunoistoquímica. O fenótipo gastrointestinal tem sido pouco explorado em relação a possíveis diferenças entre os padrões de diferenciação gástrico e intestinal, desde que os dois são geralmente analisados juntos. Os tumores mucinosos borderline, considerado um estágio precoce da carcinogênese dos tumores mucinosos, são freqüentemente associados com pseudomixoma peritoneal (PMP), o qual foi recentemente relacionado a neoplasias mucinosas do apêndice cecal. O propósito deste estudo foi analizar os diferentes padrões morfológicos de apresentação dos tumores mucinosos do ovário e sua associação com o potencial de malignidade e o perfil imunoistoquímico. MATERIAL E MÉTODOS: Este estudo retrospectivo incluiu 72 tumores de 63 pacientes com diagnóstico patológico presumido de tumor mucinoso primário de ovário selecionados dos arquivos da Divisão de Patologia Cirúrgica da Faculdade de Medicina da Universidade de São Paulo, de 1996 a 2005. Todos as lâminas da população de pacientes foram revisadas e classificadas de acordo com os critérios da WHO. Marcação imunoistoquímica para produtos do gene de mucina (MUC1, MUC2, MUC5AC e MUC6), RE, RP, CK7, CK20, CA19.9 e CA125 foram feitos em tissue microarrays. RESULTADOS: Nossos resultados mostraram 28 tumores benignos, 35 borderline e 9 malignos distribuídos nos fenótipos: pilórico (11), intestinal (30), gastrointestinal (20), mülleriano (4) e misto (gastrointestinal e mülleriano) (7). Seis pacientes tinham PMP associados. O estudo imunoistoquímico foi realizado em 67 tumores. Os tumores pilóricos apresentaram-se mais freqüentemente como tumores benignos (72.7%) e tiveram um perfil imunoistoquímico diferente de MUC2 (p= 0.003) e CA19.9 (p= 0.04) quando comparado com o fenótipo intestinal. MUC1 foi mais expresso entre os tumores com diferenciação mülleriana (pura ou mista) (100%, p= 0.02) quando comparado aos tumores de outros fenótipos. Os receptores hormonais foram positivos somente no fenótipo mülleriano. Os tumores borderline foram mais freqüentes nos fenótipos intestinal e gastrointestinal (37.1% e 40%), e estavam associados a PMP em 25% dos casos. Todos os tumores ovarianos associados a PMP eram de tipo histológico borderline e com fenótipo intestinal. O perfil dos tumores borderline de tipo intestinal, mesmo nos casos sem PMP, foi distinto dos outros tumores mucinosos de tipo intestinal e caracterizado pela expressão XVII de MUC2 e CK20. A média de idade das pacientes com tumores borderline de tipo intestinal sem PMP foi menor que daquelas com PMP. CONCLUSÃO: O subgrupo de tumores mucinosos de ovário de fenótipo gastrointestinal é o mais freqüente, mas é hetetogênio e composto por uma população de células de tipos pilórico e intestinal que diferem entre si em relação ao potencial de malignidade e perfil imunoistoquímico. Os tumores de fenotipo intestinal são mais freqüentemente malignos e borderline. Os tumores ovarianos associados com PMP e provavelmente também a maioria dos tumores borderline de fenótipo intestinal, mesmo sem PMP, devem ser considerados como tumores secundários, quando uma origem em apêndice cecal parece a mais provável. / NTRODUCTION: Primary ovarian neoplasms of mucinous type carry many controversial points regarding their pattern of differentiation, classified as benign, borderline and malignant tumors. They are also classified into different morphological phenotypes, recently called as gastrointestinal and seromucinous. Their heterogeneity has produced not only difficulty into morphological classification and differential diagnostic with metastatic neoplasms, but also on understanding the pathogenesis and immunohistochemical interpretation. The gastrointestinal phenotype has been little explored with respect to possible differences between the gastric and intestinal morphological patterns of differentiation, since the two have generally been analyzed together. The mucinous borderline tumors, thought to be an intermediary stage of mucinous carcinogenesis, were frequently associated with pseudomyxoma peritonei (PMP), which was recently linked to appendiceal mucinous neoplasms. The purpose of the study was to analyze the different morphological patterns of presentation of mucinous ovarian tumors and their association with malignant potential and immunohistochemical profile. MATERIAL AND METHODS: This retrospective study included 72 tumors from 63 patients with pathological diagnosis of presumed primary mucinous ovarian tumor selected from the files of the Division of Surgical Pathology of University of Sao Paulo Medical School, from 1996 to 2005. All slides from the patient cohort were reviewed and classified according to WHO criteria. Immunohistochemical staining for mucin genes products (MUC1, MUC2, MUC5AC and MUC6), ER, PR, CK7, CK20, CA19.9 and CA125 were performed in tissue microarrays. RESULTS: Our results showed 28 benign, 35 borderline and 9 malignant tumors distributed in phenotypes: pyloric (11), intestinal (30), gastrointestinal (20), müllerian (4) and mixed (7) gastrointestinal and müllerian). Six patients had PMP associated. The immunohistochemical study was performed in 67 tumors. The pyloric tumors presented more frequently as benign tumors (72.7%) and had a differential immunohistochemical profile of MUC2 (p= 0.003) and CA19.9 (p= 0.04) when compared with intestinal phenotype. MUC1 was more expressed between tumors with müllerian differentiation (pure or mixed) (100%, p= 0.02) when compared with the others. The hormonal receptors were positive only in müllerian phenotype. Borderline tumors were more frequently of intestinal and gastrointestinal phenotypes (37.1% and 40%), and were associated to PMP in 25% of the cases. All ovarian tumors associated to PMP were of borderline histology and of intestinal type. The profile of intestinal borderline tumors, even in cases without PMP, was distinct from that of other primary mucinous tumors of the intestinal type and characterized by MUC2 and CK20 expression. The median age of patients with intestinal borderline tumors without PMP was lower than those with PMP. XIX CONCLUSION: The gastrointestinal subgroup of mucinous ovarian tumors is the more frequent, but it is heterogeneous and composed of pyloric, and intestinal cell population that differ regarding malignant potential and immunoprofile. The intestinal tumors are more frequently malignant and borderline. Ovarian tumors associated with PMP and probably most intestinal borderline tumors, even without PMP, should be considered as secondary tumor when the appendiceal origin seems the most probable.

L'arthrose (OA) est une maladie articulaire chronique qui résulte de changements complexes dans le phénotype des chondrocytes. La présence de microcristaux contenant du phosphate dans les zones de cartilage lésées suggère que le métabolisme phosphocalcique contribue en partie aux modifications du phénotype chondrocytaire au cours de la maladie. De nombreuses études ont montré que des concentrations élevées en Phosphate Inorganique extracellulaire (ePi) ou en PyroPhosphate Inorganique (ePPi) ont respectivement un effet activateur ou répressif sur la minéralisation du cartilage articulaire. Comme le Fibroblast Growth Factor 23 (FGF23) régule les concentrations de Pi, FGF23 semble être un candidat aux modifications phénotypiques observées dans l'OA. De plus, il a récemment été mis en évidence que l’ePPi prévient la dédifférenciation in vitro des chondrocytes articulaires chez le rat, un effet provoqué par la production de PPi par la protéine Ank. Cela suggère que l’ePPi pourrait être un candidat pour prévenir les modifications du phénotype chondrocytaire. Premièrement, nous avons montré que l’expression de FGF23 est plus importante dans du cartilage lésé que dans du cartilage sain. Sous stimulation croissante de FGF23, les chondrocytes humains OA présentent une expression soutenue des marqueurs d’hypertrophie tels que COL10A1, VEGF et MMP13. Nous avons également démontré que l’expression de MMP13 est fortement dépendante de FGFR1 mais indépendante de Klotho et qu’elle est fortement régulée par la voie MEK/ERK et dans une moindre mesure par la voie PI3K/AKT. Deuxièmement, nous avons montré que FGF23 est produit de façon plus importante au cours de la différenciation des ATDC5 et qu’une stimulation par FGF23 augmente la minéralisation et l’expression des marqueurs d’hypertrophie, et ce, d’autant plus fortement en présence d’une stimulation par du Pi dans ces cellules. Dans la seconde partie, nous avons montré que des chondrocytes humains OA stimulés par du PPi présentent une expression diminuée des composants collagéniques de la matrice et une expression augmentée des MMPs, de la fibronectine et des intégrines. Une stimulation par le PPi active de façon importante la voie p38 et dans une moindre mesure la voie ERK pour réguler l’expression de ses gènes cibles et notamment MMP13 d’une manière Ank indépendante. Enfin, nous avons démontré qu’une stimulation par FGF23 entraine une augmentation de l’expression de Pit-1, ENPP1 et ANK ainsi que la production de PPi par les chondrocytes humains OA. Les résultats obtenus dans cette étude démontrent que le FGF23 permet localement une différenciation des chondrocytes OA vers un phénotype hypertrophique et peut potentiellement être considéré comme un facteur aggravant de l’OA. Contrairement aux données préliminaires chez le rat, le PPi permet un remodelage matriciel des chondrocytes humains OA et pourrait potentiellement contribuer aux effets pro-hypertrophiques du FGF23 / Osteoarthritis (OA) is the most common form of chronic joint disease, characterized by cartilage degeneration that results from complex changes in the chondrocyte phenotype. The presence of phosphate-containing microcrystals in the injured cartilage areas suggests the contribution of the phosphocalcic metabolism in the phenotype switch of chondrocytes during the disease. Numerous studies have shown that elevated concentrations of extracellular inorganic phosphate (ePi) or inorganic pyrophosphate (ePPi) have, respectively, activating or repressive mineralizing effects on articular cartilage. As Fibroblast Growth Factor 23 (FGF23) plays a major role in regulating concentrations of Pi, FGF23 is an attractive candidate to participate in the phenotype switch of the articular chondrocyte observed in OA. Moreover, we recently demonstrated that ePPi also prevents the in vitro dedifferentiation of articular chondrocyte in rats, an effect mostly triggered by Ank-induced release of PPi. This suggests that PPi may be an attractive candidate to prevent the phenotype switch of the articular chondrocyte. Firstly, we showed that FGF23 expression was higher in OA samples than in healthy one. When stimulated with increasing concentrations of FGF23, human OA chondrocytes displayed a sustained expression of markers of hypertrophy such as COL10A1, VEGF and MMP13. We demonstrated further, that MMP13 expression was mainly dependent on FGFR1 and independent of Klotho and was strongly regulated by the MEK/ERK cascade and to a lesser extent by the PI-3K/AKT pathway. Secondly, we showed that FGF23 and FGFRs were produced more importantly during ATDC5 differentiation and that FGF23 stimulation increased hypertrophic markers expression and mineralization in a synergic manner with Pi. In the second part, we showed that human OA chondrocytes stimulated with PPi displayed a decreased expression of collagen components of the matrix and sustained expression of MMPs, fibronectin and integrins. We demonstrated further that PPi stimulation mostly activates p38 pathway and to a lesser extent ERK pathway to regulate the expression of its target genes in an Ank-independent manner. Finally, we demonstrated that FGF23 stimulation increased Pit-1, ENPP1 and Ank expressions and PPi production by human OA chondrocyte. Altogether, the results obtained in this study demonstrate that FGF23 locally promotes differentiation of OA chondrocytes towards a hypertrophic phenotype and may therefore be considered as an aggravating factor for OA. In contrast to previous data obtained in rats, we demonstrated that PPi promotes matrix-remodeling of human OA chondrocytes and might contribute to FGF23 pro-hypertrophic effect

Crop improvement is necessary for food security as the global population is expected to exceed 9 billion by 2050. Limitations in water resources and more frequent droughts and floods will make it increasingly difficult to manage agricultural resources and increase yields. Therefore, we must improve our ability to monitor agronomic research plots and use the information they provide to predict impacts of moisture stress on crop growth and yield. Towards this end, agronomists have used reductions in leaf expansion rates as a visible ‘plant-based’ indicator of moisture stress. Also, modeling researchers have developed crop models such as AquaCrop to enable quantification of the severity of moisture stress and its impacts on crop growth and yield. Finally, breeders are using Unmanned Aircraft Systems (UAS) in field-based High-Throughput Phenotyping (HTP) to quickly screen large numbers of small agronomic research plots for traits indicative of drought and flood tolerance. Here we investigate whether soybean canopy cover and color time series from high-resolution UAS ortho-images can be collected with enough spatial and temporal resolution to accurately quantify and differentiate agronomic research plots, pinpoint the timing of the onset of moisture stress, and constrain crop models such as AquaCrop to more accurately simulate the timing and severity of moisture stress as well as its impacts on crop growth and yield. We find that canopy cover time series derived from multilayer UAS image ortho-mosaics can reliably differentiate agronomic research plots and pinpoint the timing of reductions in soybean canopy expansion rates to within a couple of days. This information can be used to constrain the timing of the onset of moisture stress in AquaCrop resulting in a more realistic simulation of moisture stress and a lower likelihood of underestimating moisture stress and overestimating yield. These capabilities will help agronomists, crop modelers, and breeders more quickly develop varieties tolerant to moisture stress and achieve food security.