Ponto de colheita ideal de mangas 'Tommy Atkins' destinadas ao mercado europeu

Este trabalho foi desenvolvido em um pomar comercial do Pólo Agrícola Mossoró-Assu, localizado no município de Assu-RN. Teve como objetivo caracterizar e determinar o melhor estádio de maturação para a colheita de mangas 'Tommy Atkins' destinadas ao mercado Europeu. Os frutos foram marcados e avaliados aos 82 (E1), 89 (E2), 96 (E3) e 103 (E4) dias após a floração plena (DFP) e no estádio de colheita comercial (EC)., As avaliações foram feitas por ocasião da colheita e após 21 dias de armazenamento refrigerado (13 ± 1° C e 99 % UR). As variáveis analisadas foram cor da casca e da polpa do fruto, perda de massa, firmeza, sólidos solúveis totais (SST), acidez total titulável (ATT), relação SST/ATT, açúcares solúveis totais (AST), amido e atividade respiratória. Observou-se que, os picos climatéricos ocorreram aos 10 dias após a colheita nos frutos colhidos no estádio E2 e aos 7 dias nos colhidos nos estádios E3 e EC. Os frutos colhidos nos estádios E2, E3, E4 e EC amadureceram normalmente após 21 dias de armazenamento refrigerado. Sendo que os colhidos nos estádios E3 e EC se apresentaram muito semelhantes e alcançaram melhor qualidade, indicando que a colheita pode ser realizada nesses estádios, e que o período de 96 DFP pode ser considerado como mais um indicador do ponto de colheita.

Replicating adenoviruses targeting tumours with constitutive activation of the Wnt signalling pathway

Abstract Activation of the Wnt pathway through mutation of the adenomatous polyposis coli and 13-catenin genes is a hallmark of colon cancer. These mutations lead to constitutive activation of transcription from promoters containing binding sites for Tcf/LEF transcription factors. Tumour-selective replicating oncolytic viruses are promising agents for cancer therapy. They can in principle spread throughout a tumour mass until all the cancerous cells are killed, and clinical trials have shown that they are safe except at very high doses. Adenoviruses are interesting candidates for virotherapy because their biology is well understood and their small genome can be rapidly mutated. Adenoviruses with Tcf binding sites in the E2 early promoter replicate selectively in cells with an active Wnt pathway. Although these viruses can replicate in a broad panel of colon cancer cell lines, some colorectal cancer cells are only semi-permissive for Tcf-virus replication. The aim of my thesis was to increase the safety and the efficacy of Tcf-viruses for colon cancer virotherapy. I replaced the endogenous ElA viral promoter by four Tcf binding sites and showed that transcription from the mutant promoter was specifically activated by the Wnt pathway. A virus with Tcf binding sites in the ElA and E4 promoters was more selective for the Wnt pathway than the former Tcf-E2 viruses. Moreover, insertion of Tcf binding sites into all early promoters further increased viral selectivity, but reduced viral activity. I showed that Tcf-dependent transcription was inhibited by the interaction between ElA and p300, but deletion of the p300-binding site of ElA generally led to viral attenuation. In the semi-permissive cell lines, replication of Tcf-viruses remained lower than that of the wild-type virus. The E2 promoter was the most sensitive to the cell type, but I was unable to improve its activity by targeted mutagenesis. To increase the toxicity of the Tcf-E1A/E4 virus, I decided to express a suicide gene, yeast cytosine deaminase (yCD), late during infection. This enzyme converts the prodrug 5-FC to the cytotoxic agent 5-FU. yCD was expressed in a DNA replication-dependent manner and increased viral toxicity in presence of 5-FC. Tcf-ElA and yCD adenoviruses are potentially useful vectors for the treatment of liver metastases from colorectal tumours. Résumé Dans la quasi-totalité des cancers du côlon, la voie Wnt est activée par des mutations dans les gènes codant pour APC ou pour la (3-caténine. Ces mutations activent de façon constitutive la transcription de promoteurs contenant des sites de liaison pour les facteurs de transcription Tcf/LEF. Les virus réplicatifs spécifiques aux tumeurs sont des agents prometteurs pour la thérapie cancéreuse. En principe, ces vecteurs peuvent se propager dans une masse tumorale jusqu'à destruction de toutes les cellules cancéreuses, et des études cliniques ont démontré que de tels vecteurs n'étaient pas toxiques, sauf à de très hautes doses. Les adénovirus sont des candidats intéressants pour la thérapie virale car leur biologie est bien définie et leur petit génome peut être rapidement modifié. Des adénovirus comportant des sites de liaison à Tcf dans leur promoteur précoce E2 se répliquent sélectivement dans les cellules qui possèdent une voie Wnt active. Ces virus sont capables de se répliquer dans un grand nombre de cellules cancéreuses du côlon, bien que certaines de ces cellules ne soient que semi-permissives pour la réplication des virus Tcf. Le but de ma thèse était d'augmenter la sécurité et l'efficacité des virus Tcf. Le promoteur viral endogène ElA a été remplacé par quatre sites de liaison à Tcf, ce qui a rendu son activation dépendante de la voie Wnt. Un virus comportant des sites de liaison pour Tcf dans les promoteurs ElA et E4 était plus sélectif pour la voie Wnt que les précédents virus Tcf-E2, et un virus comportant des sites Tcf dans tous les promoteurs précoces était encore plus sélectif, mais moins actif. J'ai montré que l'interaction entre ElA et p300 inhibait la transcription dépendante de Tcf, mais la délétion du domaine concerné dans ElA a eu pour effet d'atténuer les virus. Dans les cellules semi-permissives, la réplication des virus Tcf était toujours plus basse que celle du virus sauvage. J'ai identifié le promoteur E2 comme étant le plus sensible au type cellulaire, mais n'ai pas pu augmenter son activité par mutagenèse. Pour augmenter la toxicité du virus Tcf-E1A/E4, j'ai décidé d'exprimer un gène suicide, la cytosine déaminase (yCD), pendant la phase tardive de l'infection. Cette enzyme transforme la procirogue 5-FC en l'agent cytotoxique 5-FU. yCD était exprimée après réplication de l'ADN viral et augmentait la toxicité virale en présence de 5-FC. Les virus Tcf-ElA et yCD sont des vecteurs potentiellement utiles pour le traitement des métastases hépatiques de cancers colorectaux.

Functions of the human papillomavirus type 16 E4 protein

1. Abstract Cervical cancer is thought to be the consequence of infection by human papillomaviruses (HPV). In the majority of cases, DNA from HPV type 16 (HPV16) is found in malignant cervical lesions. The initial steps leading to transformation of an infected cell are not clearly understood but in most cases, disruption and integration of the episomal viral DNA must take place. As a consequence, the E2 and E4 genes are usually not expressed whereas the E6 and E7 oncogenes are highly expressed. However, in a normal infection in which the viral DNA is maintained as an episome, all viral genes are expressed. The pattern according to which the viral proteins are made, and therefore the life cycle of the virus, is tightly linked to the differentiation process of the host keratinocyte. The study of the viral oncogenes E6 and E7 has revealed crucial functions in the process of malignant transformation such as degradation of the p53 tumor suppressor protein, deregulation of the Retinoblastoma protein pathway and activation of the telomerase ribonucleoprotein. All these steps are necessary for cancerous lesions to develop. However, the loss of the E2 gene product seems to be necessary for sufficient expression of E6 and E7 in order to achieve such effects. In normal infections, the E4 protein is made abundantly in the later stages of the viral life cycle. Though extensive amounts of work have been carried out to define the function of E4, it still remains unclear. In this study, several approaches have been used to try and determine the functions of E4. First, a cell-penetrating fusion protein was designed and produced in order to circumvent the chronic difficulties of expressing E4 in mammalian cells. Unfortunately, this approach was not successful due to precipitation of the purified fusion protein. Second, the observation that E4 accumulates in cells having modified their adhesion properties led to the hypothesis that E4 might be involved in the differentiation process of keratinocytes. Preliminary results suggest that E4 triggers differentiation. Last, as E4 has been reported to collapse the cytokeratin network of keratinocytes, a direct approach using atomic force microscopy has allowed us to test the potential modification of mechanical properties of cells harboring reorganized cytokeratin networks. If so, a potential role for E4 in viral particle release could be hypothesized. 2. Résumé Il a été établi que le cancer du col de l'utérus se développe essentiellement à la suite d'une infection par le virus du papillome humain (HPV). Dans la majorité des cas analysés, de l'ADN du HPV de type 16 (HPV16) est détecté. Les étapes initiales de la transformation d'une cellule infectée sont mal connues mais il semble qu'une rupture du génome viral, normalement épisomal, suivi d'une intégration dans le génome de la cellule hôte soient des étapes nécessaires dans la plupart des cas. Or il semble qu'il y ait une sélection pour les cas où l'expression des oncogènes viraux E6 et E7 soit favorisée alors que l'expression des gènes E2 et E4 est en général impossible. Par contre, dans une infection dite normale où le génome viral n'est pas rompu, il n'y pas développement de cancer et tous les gènes viraux sont exprimés. L'ordre dans lequel les protéines virales sont produites, et donc le cycle de réplication du virus, est intimement lié au processus de différentiation de la cellule hôte. L'étude des protéines oncogènes E6 et E7 a révélé des fonctions clés dans le processus de transformation des cellules infectées telles que la dégradation du suppresseur de tumeur p53, la dérégulation de la voie de signalisation Rb ainsi que l'activation de la télomérase. Toutes ces activités sont nécessaires au développement de lésions cancéreuses. Toutefois, il semble que l'expression du gène E2 doit être empêchée afin que suffisamment des protéines E6 et E7 soient produites. Lorsque le gène E2 est exprimé, et donc lorsque le génome viral n'est pas rompu, les protéines E6 et E7 n'entraînent pas de telles conséquences. Le gène E4, qui se trouve dans la séquence codante de E2, a aussi besoin d'un génome viral intact pour être exprimé. Dans une infection normale, le gène E4 est exprimé abondamment dans les dernières étapes de la réplication du virus. Bien que de nombreuses études aient été menées afin de déterminer la fonction virale à E4, aucun résultat n'apparaît évident. Dans ce travail, plusieurs approches ont été utilisées afin d'adresser cette question. Premièrement, une protéine de fusion TAT-E4 a été produite et purifiée. Cette protéine, pouvant entrer dans les cellules vivantes par diffusion au travers de la membrane plasmique, aurait permis d'éviter ainsi les problèmes chroniques rencontrés lors de l'expression de E4 dans les cellules mammifères. Malheureusement, cette stratégie n'a pas pu être utilisée à cause de la précipitation de la protéine purifiée. Ensuite, l'observation que E4 s'accumule dans les cellules ayant modifié leurs propriétés d'adhésion a suggéré que E4 pourrait être impliqué dans le procédé de différentiation des kératinocytes. Des résultats préliminaires supportent cette possibilité. Enfin, il a été montré que E4 pouvait induire une réorganisation du réseau des cytokératines. Une approche directe utilisant le microscope à force atomique nous a ainsi permis de tester une potentielle modification des propriétés mécaniques de cellules ayant modifié leur réseau de cytokératines en présence de E4. Si tel est le cas, un rôle dans la libération de particules virales peut être proposé pour E4.

El rasgo de personalidad "Búsqueda de sensaciones": una teoría psicobiológica

En este trabajo se expone y se sintetiza la teoría del rasgo de personalidad denominado "búsqueda de sensaciones", se proporciona información cronológica de la investigación empírica realizada desde los años 60 hasta nuestros días. Se incide en los correlatos biológicos de este rasgo en particular Potenciales Evocados (PE), Monoaminoxidades Plaquetear (MAO), Reflejo Orienativo (RO) y hormonas sexuales como la Testosterona (T) y Estradiol (E2). Se sugiere que se avance en la investigación psicobiológica del rasgo en cuestión, esfuerzo que por otro lado está generando un importante soporte empírico a la teoría.

Efeitos do ácido giberélico e do thidiazuron sobre as características dos frutos e do mosto da uva 'Niagara Rosada'

Com o objetivo de avaliar os efeitos do ácido giberélico e thidiazuron nas características dos cachos de uvas cultivar Niagara Rosada, foram conduzidos dois experimentos. No primeiro ensaio, utilizaram-se soluções aquosas de ácido giberélico (AG3) 0; 15; 30; 45; 60; 75 e 90mg.L-1, aplicadas no florescimento e repetidas aos 14 dias após (E1E2) e no outro tratamento, o mesmo composto e doses aplicados uma única vez, aos 14 dias após o florescimento (E2); no segundo experimento, thidiazuron (TDZ) 0; 5; 10; 15; 20; 25 e 30 mg.l-1, aplicados quatro dias antes da antese e repetidos aos seis dias após o florescimento (E1E2); e no outro tratamento, o mesmo composto e doses aplicados uma única vez, aos seis dias após o florescimento (E2). Pelos resultados obtidos, verificou-se que os tratamentos com thidiazuron aumentaram a fixação das bagas, resultando em incremento da massa dos cachos e o engrossamento significativo do engaço. Os frutos tratados com thidiazuron apresentaram maturação desuniforme, bagas com massa e tamanho reduzidos, cachos bastante compactos, com baixo valor comercial. Aumentos no número de bagas com incrementos na massa dos cachos também foram observados com aplicações de ácido giberélico. Para a variável massa dos cachos os melhores resultados obtidos pela estimativa da equação de regressão foram verificados com aplicações de ácido giberélico a 54mg.L-1, com valores de 198,8g.

Reguladores vegetais influenciando número e tamanho de células das bagas da uva 'Niagara Rosada'

O objetivo do trabalho foi avaliar o efeito do ácido giberélico, thidiazuron e quinmerac na anatomia das bagas de uvas cultivar 'Niagara Rosada', provenientes de vinhedo localizado em Dourados - MS. Realizaram-se três experimentos. No primeiro ensaio, utilizaram-se soluções aquosas de ácido giberélico (AG3) 0; 15; 30; 45; 60; 75 e 90 mg.l-1, aplicadas no florescimento e repetidas aos 14 dias após (E1E2) e, no outro tratamento, o mesmo composto e doses aplicados uma única vez aos 14 dias após o florescimento (E2); no segundo experimento, thidiazuron (TDZ) 0; 5; 10; 15; 20; 25 e 30 mg.l-1, aplicados quatro dias antes da antese e repetidos aos seis dias após o florescimento (E1E2); e no outro tratamento, o mesmo composto e doses aplicados uma única vez aos seis dias após o florescimento (E2); e, no terceiro, quinmerac 0; 10; 20; 30; 40; 50 e 60 mg.l-1, aplicados no florescimento e repetidos aos 14 dias após (E1E2), e, no outro tratamento, o mesmo composto e doses aplicados uma única vez, 14 dias após o florescimento (E2). As variáveis avaliadas foram: número e dimensões das células das bagas. Pelos resultados obtidos, verificou-se que duas aplicações de ácido giberélico, thidiazuron e quinmerac promoveram a divisão celular, enquanto a expansão celular foi observada com uma única aplicação de thidiazuron e quinmerac.

Clonagem do abacateiro variedade "Duke 7" (Persea americana Mill.) por alporquia

Foram conduzidos dois experimentos com a finalidade de determinar a possibilidade de clonagem da variedade de abacateiro "Duke 7", por alporquia e a influência do AIB (ácido indol-3-butírico) no processo. Experimento 1 - Alporque em plantas - O delineamento experimental utilizado foi o inteiramente casualizado, em esquema fatorial 2L x 4N x 2E, correspondendo à manutenção ou não das plantas à ausência de luz (L), níveis de AIB (N) e tipo de estrutura (E). Nos quatro dias antecedentes à realização da alporquia, 50% das plantas permaneceram na ausência total de luz (L1), e as demais, em condições normais de ripado, 50% de luminosidade (L2). No local anelado, foram aplicadas as concentrações (N) de AIB (ácido indolbutírico): 0; 1.000, 3.000 e 5.000 mg kg-1. O experimento foi realizado em duas estruturas diferenciadas pelo tipo de cobertura: a estrutura um (E1) e a estrutura dois (E2), diferenciadas pela temperatura e intensidade luminosa. Experimento 2- Alporque em plantas adultas após poda drástica - Os alporques foram realizados cinco meses após a poda drástica, quando os ramos possuíam entre 1,5 e 2,0 cm de diâmetro. O delineamento experimental utilizado foi o inteiramente casualizado em esquema fatorial, com 4 tratamentos, caracterizados pelas concentrações de AIB (0; 1.000; 3.000 e 5.000 mg kg-1), com quatro repetições, e cada parcela composta por 10 alporques. Em nenhum dos experimentos, houve enraizamento dos alporques, consequentemente há necessidade de maiores estudos, quanto à clonagem da variedade "Duke 7" para viabilizá-la como porta-enxerto.

PPARβ/δ ameliorates fructose-induced insulin resistance in adipocytes by preventing Nrf2 activation

We studied whether PPARβ/δ deficiency modifies the effects of high fructose intake (30% fructose in drinking water) on glucose tolerance and adipose tissue dysfunction, focusing on the CD36-dependent pathway that enhances adipose tissue inflammation and impairs insulin signaling. Fructose intake for 8weeks significantly increased body and liver weight, and hepatic triglyceride accumulation in PPARβ/δ-deficient mice but not in wild-type mice. Feeding PPARβ/δ-deficient mice with fructose exacerbated glucose intolerance and led to macrophage infiltration, inflammation, enhanced mRNA and protein levels of CD36, and activation of the JNK pathway in white adipose tissue compared to those of water-fed PPARβ/δ-deficient mice. Cultured adipocytes exposed to fructose also exhibited increased CD36 protein levels and this increase was prevented by the PPARβ/δ activator GW501516. Interestingly, the levels of the nuclear factor E2-related factor 2 (Nrf2), a transcription factor reported to up-regulate Cd36 expression and to impair insulin signaling, were increased in fructose-exposed adipocytes whereas co-incubation with GW501516 abolished this increase. In agreement with Nrf2 playing a role in the fructose-induced CD36 protein level increases, the Nrf2 inhibitor trigonelline prevented the increase and the reduction in insulin-stimulated AKT phosphorylation caused by fructose in adipocytes. Protein levels of the well-known Nrf2 target gene NAD(P)H: quinone oxidoreductase 1 (Nqo1) were increased in water-fed PPARβ/δ-null mice, suggesting that PPARβ/δ deficiency increases Nrf2 activity; and this increase was exacerbated in fructose-fed PPARβ/δ-deficient mice. These findings indicate that the combination of high fructose intake and PPARβ/δ deficiency increases CD36 protein levels via Nrf2, a process that promotes chronic inflammation and insulin resistance in adipose tissue.

Diferentes espaçamentos de plantio e níveis de adubação sobre a atividade da redutase do nitrato em folhas do híbrido de mamoeiro UENF/CALIMAN-01

O objetivo deste trabalho foi avaliar o efeito de diferentes espaçamentos de plantio e de níveis de adubação NPK sobre a atividade da redutase do nitrato (RN) nas folhas do híbrido de mamoeiro UENF/CALIMAN-01 , visando a sugerir possível ajuste em seu manejo de adubação nitrogenada, no sentido de maximizar a eficiência do uso do nitrogênio. O experimento foi conduzido na fazenda Caliman Agrícola S.A., no município de Linhares - ES. Utilizou-se o delineamento estatístico experimental em blocos casualizados, com esquema fatorial, com três espaçamentos de plantio entre plantas (E1 = 1,8 m; E2 = 2,25 m, e E3 = 2,7 m), cinco níveis de adubação NPK convencional (A1 = 80% do padrão; A2 = 100% padrão da empresa; A3 = 120% do padrão; A4 = 140% do padrão, e A5 = 160% do padrão) e cinco períodos de avaliação (meses de março a julho). O padrão de adubação NPK da empresa consiste em 350; 105 e 660 kg ha-1ano-1 de sulfato de amônio (20% de N), superfosfato simples (18% de P) e cloreto de potássio (60% de K), respectivamente. Os dados obtidos para a atividade da RN foram submetidos a uma análise de variância e teste de médias. Dentre os tratamentos testados, o nível A1 (80% do padrão), independentemente do espaçamento, poderia ser indicado no manejo do híbrido de mamoeiro UENF/CALIMAN-01, pois em todos eles a atividade da redutase do nitrato, em praticamente todos os períodos avaliados, apresentou valores adequados, ou até mesmo superiores aos encontrados na literatura em cultivares de mamoeiro. A redução da adubação NPK pôde ser justificada, uma vez que não houve diferença na produtividade das plantas entre os tratamentos avaliados.

Comportamento produtivo da videira, cultivar Juliana, sobre três porta-enxertos em diferentes épocas de poda

Este trabalho objetivou verificar a influência de três porta-enxertos no comportamento da cultivar Juliana em relação à fenologia, em três épocas de poda e em relação aos caracteres físicos de cachos, de bagas e de engaços, em duas épocas de poda. Os ensaios foram avaliados em Jundiaí-SP, e as podas foram realizadas em 15-08-2007, 22-01-2008 e 16-09-2008. O delineamento experimental foi o em blocos inteiramente casualizados, com parcelas subdivididas, com três repetições, sendo as parcelas representadas pela combinação da cultivar Juliana enxertada sobre os porta-enxertos 'Ripária do Traviú', 'IAC 572' e 'IAC766', e as subparcelas, pelas épocas de poda, que corresponderam a três para os estádios fenológicos e a duas para os caracteres físicos de cachos, bagas e engaço. Não houve diferença entre os porta-enxertos para os estádios fenológicos e para os caracteres físicos de cachos e de bagas, com exceção para massa da matéria fresca de baga e massa fresca de engaço. Maior massa da matéria fresca de engaço foi obtida pela combinação da cultivar Juliana e o porta-enxerto 'IAC 572'. Para as diferentes épocas de poda, foram detectadas diferenças para todos os estádios fenológicos, havendo interação entre porta-enxerto e época de poda para os estádios E1 - período da poda ao início da brotação; E2 - período da poda ao início do florescimento, e E5 - período da poda ao início da colheita.

Manejo da cultura do híbrido de mamoeiro (Carica papaya L.) do grupo 'Formosa' UENF/CALIMAN : 01 para melhoria na qualidade do fruto com menor aplicação de adubação NPK

O presente trabalho teve como objetivo avaliar o efeito de alguns aspectos do manejo da cultura do mamoeiro como espaçamento e nível de adubação NPK, sobre alguns atributos de qualidade dos frutos do híbrido do grupo 'Formosa' UENF/CALIMAN-01(UC01). O experimento foi conduzido na fazenda Caliman Agrícola S.A., em Linhares-ES. Utilizou-se o delineamento estatístico experimental em blocos casualizados, com esquema fatorial, com três espaçamentos de plantio entre plantas (E1 = 1,80 m; E2 = 2,25 m, e E3 = 2,70 m), cinco níveis de adubação NPK convencional (A1 = 80% do padrão da empresa (PE); A2 = 100% PE; A3 = 120% PE; A4 = 140% PE, e A5 = 160% PE), e três períodos de avaliação (junho, agosto e outubro de 2007). O padrão de adubação NPK da empresa (PE) consiste em 350; 105 e 660 kg ha-1ano-1 de sulfato de amônio (20% de N), superfosfato simples (18% de P2O5) e cloreto de potássio (60% de K2O), respectivamente. Foram analisados a firmeza do fruto e da polpa, a concentração de sólidos solúveis (SS), o pH, a acidez titulável (AT) e a razão SS/AT da polpa. Os dados foram submetidos à análise de variância e teste de médias. Os resultados mostram que, entre as condições de espaçamento e níveis de adubação NPK testados, o melhor desempenho foi obtido pelas combinações E1A1 ou E2A1, os quais devem ser adotadas para o manejo do híbrido UC01. Os tratamentos resultaram em frutos com atributos de qualidade superiores, além de proporcionar redução nos gastos com adubação NPK e menor impacto ambiental em função da aplicação excessiva de adubo no solo.

Mini invasive skeletal muscle biopsy technique with a tri-axial end cut needle.

OBJECTIVE: Skeletal Muscle Biopsy is a minor surgical procedure for the diagnosis of different neuromuscular pathological conditions and has recently gained popularity also in the research field of age-related muscular modifications and sarcopenia. Few studies focused on the application of mini-invasive muscular biopsy in both normal and pathological conditions. The aim of our study was to describe a mini invasive ultrasound-guided skeletal muscular biopsy technique in complete spinal cord injured (SCI) patients and healthy controls with a tri-axial end-cut needle. PATIENTS AND METHODS: Skeletal muscle biopsies were collected from 6 chronic SCI patients and 3 healthy controls vastus lateralis muscle with a tri-axial end cut needle (Biopince© - Angiotech). Muscle samples were stained for ATPase to determine fibers composition, moreover, gene expression of cyclooxygenase-1 (COX-1) and prostaglandin E2 receptor has been analyzed by Real Time RT-PCR. RESULTS: All the procedures were perfomed easily without failures and complications. Control tissue was macroscopically thicker than SCI one. Control specimen displayed an equal distribution of type I and type II fibers, while SCI sample displayed a prevalence of type II fibers SCI specimen displayed a significant reduction in COX-1 gene expression. This mini-invasive approach was easy, accurate and with low complication rate in performing skeletal muscle biopsy in both SCI patients and controls. CONCLUSIONS: This technique could be useful in conditions in which the overall quantity of specimen required is small like for molecular biology analysis. For histological diagnostic purposes and/or conditions in which the original tissue is already pathologically modified, this technique should be integrated with more invasive techniques.

Thyroid function in the elderly and sex hormones in elderly men: reference intervals and associations with health

Aims: The aims were to create clinically feasible reference intervals for thyroidstimulating hormone (TSH) and free thyroxine (FT4) and to analyze associations between thyroid function and self-rated health, neuropsychiatric symptoms, depression and dementia in the elderly. The second aim was also to establish reference intervals for sex hormones and to analyze associations between sex hormone levels and self-rated health, symptoms, depression and dementia in elderly men. Subjects and methods: The study population comprised 1252 subjects aged 65 years or over, living in the municipality of Lieto, south-western Finland. Self-rated health, life satisfaction, symptoms, depression, and dementia were assessed with specific questions, clinical examination and tools such as the Zung Self-report Depression Scale and the Mini-Mental State Examination. Independent variables were dichotomized, and associations of these variables with TSH, FT4 or sex hormone levels were assessed. Levels of TSH and FT4 in thyroid disease–free women and women treated with thyroxine were also compared. Results: Elevated concentrations of thyroid peroxidase antibodies (TPOAb) or thyroglobulin antibodies (TgAb) were found to have a marked effect on the upper reference limit for TSH among women, who were thyroid antibody positive more higher than suggested in several recent guidelines. After age adjustment, there were no associations between TSH levels and self-rated health, life satisfaction, or most neuropsychiatric symptoms in the thyroid disease-free population. Although women with thyroxine treatment for primary hypothyroidism had far higher TSH levels than thyroid disease-free women, there were no differences between thyroid-disease free women and women with stable thyroxine treatment regarding self-rated health, life satisfaction or symptoms. Age had a significant positive association with luteinizing hormone (LH), follicle 2 practice, one range in men aged 65 years or over can be used for T, E2 and FSH measured with the AutoDelfia method, but two separate reference intervals should be used for fT, LH and SHBG. After adjustment for age, higher levels of T and fT were associated with better self-rated health (SRH) in the reference population. After adjustment for age and body mass index (BMI), there were no associations between sex hormone concentrations and self-rated health, life satisfaction or most symptoms in concentration. Conclusion: Age-specific reference intervals were derived for thyroid function and sex hormones based on comprehensive data from a community-dwelling population with a high participation rate. The results do not support the need to decrease the upper reference limit for TSH or to lower the optimal TSH target in levothyroxine treatment in older adults, as recommended in recent guidelines. Older age or being overweight symptoms among elderly men. The associations of single symptoms with T levels were inconsistent among elderly men, although the association of low T level with diagnosed depression might be clinically significant.

p21 as a Transcriptional Co-Repressor of S-Phase and Mitotic Control Genes

It has been previously described that p21 functions not only as a CDK inhibitor but also as a transcriptional co-repressor in some systems. To investigate the roles of p21 in transcriptional control, we studied the gene expression changes in two human cell systems. Using a human leukemia cell line (K562) with inducible p21 expression and human primary keratinocytes with adenoviral-mediated p21 expression, we carried out microarray-based gene expression profiling. We found that p21 rapidly and strongly repressed the mRNA levels of a number of genes involved in cell cycle and mitosis. One of the most strongly down-regulated genes was CCNE2 (cyclin E2 gene). Mutational analysis in K562 cells showed that the N-terminal region of p21 is required for repression of gene expression of CCNE2 and other genes. Chromatin immunoprecipitation assays indicated that p21 was bound to human CCNE2 and other p21-repressed genes gene in the vicinity of the transcription start site. Moreover, p21 repressed human CCNE2 promoter-luciferase constructs in K562 cells. Bioinformatic analysis revealed that the CDE motif is present in most of the promoters of the p21-regulated genes. Altogether, the results suggest that p21 exerts a repressive effect on a relevant number of genes controlling S phase and mitosis. Thus, p21 activity as inhibitor of cell cycle progression would be mediated not only by the inhibition of CDKs but also by the transcriptional down-regulation of key genes.

Cyclooxygenase-2 Expression in Non-Small Cell Lung Cancer Correlates With Hypertrophic Osteoarthropathy.

Hypertrophic osteoarthrpathy (HO) is a rare paraneoplasic syndrome associated with non-small cell lung cancer (NSCLC). The pathophysiology of HO is unknown but was recently related to enhanced levels of urine prostaglandin E2 (PGE2). Here, we report the case of a patient that presented HO in association with a resectable left upper lobe NSCLC. Following surgery and adjuvant chemotherapy, HO resolved and did not recur with development of a brain metastasis 1 year later. Interestingly, tumor cyclooxygenase-2, an enzyme responsible the synthesis of PGE2, was expressed in the primary tumor but not in the resected metastasis.