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1.
Arch. prev. riesgos labor. (Ed. impr.) ; 18(1): 25-27, ene.-mar. 2015.
Artigo em Espanhol | IBECS | ID: ibc-131949

RESUMO

Tradicionalmente, la investigación sobre aspectos psicosociales en el sector de la construcción se ha centrado principalmente en aspectos negativos de la salud y en resultados tan negativos como los accidentes laborales. Sin embargo, este estudio se centra en las relaciones específicas entre los diferentes factores psicosociales positivos compartidos por trabajadores de la construcción que podrían ser responsables de su bienestar y de su desempeño laboral. El objetivo principal de este estudio fue evaluar si los recursos personales se relacionan con el desempeño laboral auto-percibido a través de los recursos laborales y el engagement (implicación) en el trabajo. Considerando las predicciones de la Teoría Social Cognitiva de Bandura y el proceso motivacional del Modelo Demandas-Recursos, esperamos que la relación entre los recursos personales y el desempeño estará totalmente mediada por los recursos laborales y el engagement en el trabajo. La muestra se compone de 228 trabajadores de la construcción. Modelos de ecuaciones estructurales apoyaron el modelo de investigación. Los recursos personales (concretamente la auto-eficacia, las competencias mentales y las competencias emocionales) juegan un papel predictivo en la percepción de los recursos laborales (concretamente en el control del trabajo y en el apoyo social de supervisor), que a su vez, influye en el engagement en el trabajo y en el desempeño auto-percibido. Este estudio pone de manifiesto el papel fundamental que desempeñan los recursos personales en la determinación de cómo los trabajadores perciben los recursos laborales disponibles, de los niveles de engagement en el trabajo y de su desempeño laboral auto-percibido. Se discuten las implicaciones teóricas y prácticas


Traditionally, research focussing on psychosocial factors in the construction industry has focused mainly on the negative aspects of health and on results such as occupational accidents. This study, however, focuses on the specificrelationships among the different positive psychosocial factors shared by construction workers that could beresponsible for occupational well-being and outcomes such as performance. The main objective of this study was totest whether personal resources predict self-rated job performance through job resources and work engagement. Following the predictions of Bandura's Social Cognitive Theory and the motivational process of the Job Demands-Resources Model, we expect that the relationship between personal resources and performance will be fully mediatedby job resources and work engagement. The sample consists of 228 construction workers. Structural equation modelling supports the research model. Personal resources (i.e. self-efficacy, mental and emotional competences) play a predicting role in the perception of job resources (i.e. job control and supervisor social support), which inturn leads to work engagement and self-rated performance. This study emphasises the crucial role that personalresources play in determining how people perceive job resources by determining the levels of work engagement and, hence, their self-rated job performance. Theoretical and practical implications are discussed


Assuntos
Humanos , Autoeficácia , 16360 , Acidentes de Trabalho/prevenção & controle , Processos Grupais , Impacto Psicossocial , 51829
2.
J Musculoskelet Neuronal Interact ; 14(4): 432-44, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25524969

RESUMO

OBJECTIVES: To investigate the effect of whey protein plus potassium bicarbonate supplement on disused skeletal muscle structure and proteolysis after bed rest (BR). METHODS: Soleus (SOL) and vastus lateralis (VL) biopsies were sampled from ten (n=10) healthy male subjects (aged 31±6 years) who did BR once with and once without protein supplement as a dietary countermeasure (cross-over study design). The structural changes (myofibre size and type distribution) were analysed by histological sections, and muscle protein breakdown indirectly via the proteolysis markers, calpain 1 and 3, calpastatin, MuRF1 and 2, both in muscle homogenates and by immunohistochemistry. RESULTS: BR caused size-changes in myofiber cross-sectional area (FCSA, SOL, p=0,004; VL, p=0.03), and myofiber slow-to-fast type transition with increased hybrids (SOL, p=0.043; VL, p=0.037) however with campaign differences in SOL (p<0.033). No significant effect of BR and supplement was found by any of the key proteolysis markers. CONCLUSIONS: Campaign differences in structural muscle adaptation may be an issue in cross-over design BR studies. The whey protein plus potassium bicarbonate supplement did not attenuate atrophy and fibre type transition during medium term bed rest. Alkaline whey protein supplements may however be beneficial as adjuncts to exercise countermeasures in disuse.


Assuntos
Repouso em Cama/efeitos adversos , Bicarbonatos/uso terapêutico , Proteínas do Leite/uso terapêutico , Atrofia Muscular/prevenção & controle , Compostos de Potássio/uso terapêutico , Proteólise/efeitos dos fármacos , Adulto , Estudos Cross-Over , Suplementos Nutricionais , Humanos , Imuno-Histoquímica , Masculino , Proteínas do Soro do Leite , Adulto Jovem
3.
J Musculoskelet Neuronal Interact ; 13(2): 166-77, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23728103

RESUMO

Human performance in microgravity is characterized by reversed skeletal muscle actions in terms of active vs. passive mode contractions of agonist/antagonist groups that may challenge principal biodynamics (biomechanical forces translated from muscle to bone) of the skeletal muscle-bone unit. We investigated active vs. passive muscle motions of the unloaded hindlimb skeletal muscle-bone unit in the 21 days tail-suspended (TS) rat using a newly designed stepper exercise device. The regimen included both active mode motions (TSA) and passive mode motions (TSP). A TS-only group and a normal cage group (CON) served as positive or negative controls. The muscle and bone decrements observed in TS-only group were not seen in the other groups except TSP. Active mode motions supported femur and tibia bone quality (5% BMD, 10% microtrabecular BV/TV, Tb.Th., Tb.N. parameters), whole soleus muscle/myofiber size and type II distribution, 20% increased sarcolemma NOS1 immunosignals vs. CON, with 25% more hybrid fiber formation (remodeling sign) for all TS groups. We propose a new custom-made stepper device to be used in the TS rat model that allows for detailed investigations of the unique biodynamic properties of the muscle-bone unit during resistive-load exercise countermeasure trials on the ground or in microgravity.


Assuntos
Osso e Ossos/anatomia & histologia , Osso e Ossos/fisiologia , Elevação dos Membros Posteriores/fisiologia , Músculo Esquelético/anatomia & histologia , Músculo Esquelético/fisiologia , Condicionamento Físico Animal/fisiologia , Absorciometria de Fóton , Animais , Fenômenos Biomecânicos/fisiologia , Peso Corporal/fisiologia , Osso e Ossos/diagnóstico por imagem , Eletromiografia , Imunofluorescência , Imuno-Histoquímica , Extremidade Inferior/fisiologia , Terapia Passiva Contínua de Movimento , Músculo Esquelético/diagnóstico por imagem , Óxido Nítrico Sintase Tipo I/metabolismo , Tamanho do Órgão , Ratos , Ratos Sprague-Dawley , Treinamento Resistido , Tíbia/anatomia & histologia , Tíbia/diagnóstico por imagem , Tíbia/fisiologia , Tomografia Computadorizada por Raios X
4.
J Musculoskelet Neuronal Interact ; 10(3): 207-19, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20811145

RESUMO

Long-term bed-rest is used to simulate the effect of spaceflight on the human body and test different kinds of countermeasures. The 2nd Berlin BedRest Study (BBR2-2) tested the efficacy of whole-body vibration in addition to high-load resisitance exercise in preventing bone loss during bed-rest. Here we present the protocol of the study and discuss its implementation. Twenty-four male subjects underwent 60-days of six-degree head down tilt bed-rest and were randomised to an inactive control group (CTR), a high-load resistive exercise group (RE) or a high-load resistive exercise with whole-body vibration group (RVE). Subsequent to events in the course of the study (e.g. subject withdrawal), 9 subjects participated in the CTR-group, 7 in the RVE-group and 8 (7 beyond bed-rest day-30) in the RE-group. Fluid intake, urine output and axiallary temperature increased during bed-rest (p < .0001), though similarly in all groups (p > or = .17). Body weight changes differed between groups (p < .0001) with decreases in the CTR-group, marginal decreases in the RE-group and the RVE-group displaying significant decreases in body-weight beyond bed-rest day-51 only. In light of events and experiences of the current study, recommendations on various aspects of bed-rest methodology are also discussed.


Assuntos
Repouso em Cama/efeitos adversos , Terapia por Exercício/métodos , Aptidão Física/fisiologia , Simulação de Ausência de Peso/efeitos adversos , Adulto , Berlim , Humanos , Masculino , Pessoa de Meia-Idade , Osteoporose/etiologia , Osteoporose/fisiopatologia , Osteoporose/prevenção & controle , Resultado do Tratamento , Vibração/uso terapêutico , Adulto Jovem
5.
J Anat ; 212(3): 306-18, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18221329

RESUMO

The cellular mechanisms of human skeletal muscle adaptation to disuse are largely unknown. The aim of this study was to determine the morphological and biochemical changes of the lower limb soleus and vastus lateralis muscles following 60 days of head-down tilt bed rest in women with and without exercise countermeasure using molecular biomarkers monitoring functional cell compartments. Muscle biopsies were taken before (pre) and after bed rest (post) from a bed rest-only and a bed rest exercise group (n = 8, each). NOS1 and NOS3/PECAM, markers of myofibre 'activity' and capillary density, and MuRF1 (E3 ubiquitin-ligase), a marker of proteolysis, were documented by confocal immunofluorescence and immunoblot analyses. Morphometrical parameters (myofibre cross-sectional area, type I/II distribution) were largely preserved in muscles from the exercise group with a robust trend for type II hypertrophy in vastus lateralis. In the bed rest-only group, the relative NOS1 immunostaining intensity was decreased at type I and II myofibre membranes, while the bed rest plus exercise group compensated for this loss particularly in soleus. In the microvascular network, NOS3 expression and the capillary-to-fibre ratio were both increased in the exercise group. Elevated MuRF1 immunosignals found in subgroups of atrophic myofibres probably reflected accelerated proteolysis. Immunoblots revealed overexpression of the MuRF1 protein in the soleus of the bed rest-only group (> 35% vs. pre). We conclude that exercise countermeasure during bed rest affected both NOS/NO signalling and proteolysis in female skeletal muscle. Maintenance of NO signalling mechanisms and normal protein turnover by exercise countermeasure may be crucial steps to attenuate human skeletal muscle atrophy and to maintain cell function following chronic disuse.


Assuntos
Proteínas Musculares/análise , Músculo Esquelético/metabolismo , Óxido Nítrico Sintase Tipo III/análise , Óxido Nítrico Sintase Tipo I/análise , Ubiquitina-Proteína Ligases/análise , Simulação de Ausência de Peso , Adulto , Repouso em Cama , Biomarcadores/análise , Biópsia por Agulha , Capilares/ultraestrutura , Terapia por Exercício , Feminino , Decúbito Inclinado com Rebaixamento da Cabeça , Humanos , Immunoblotting , Contração Isométrica , Microscopia Confocal , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/fisiopatologia , Atrofia Muscular/metabolismo , Atrofia Muscular/fisiopatologia , Atrofia Muscular/prevenção & controle , Músculo Quadríceps/metabolismo , Músculo Quadríceps/fisiopatologia , Tempo , Proteínas com Motivo Tripartido , Contramedidas de Ausência de Peso
6.
Rev. esp. geriatr. gerontol. (Ed. impr.) ; 41(supl.2): 2-16, sept. 2006. ilus, tab
Artigo em Espanhol | IBECS | ID: ibc-148967

RESUMO

El presente artículo aborda un proyecto de investigación-acción basado en metodología cualitativa con el propósito de fomentar un envejecimiento activo y exitoso, además de prevenir las situaciones de dependencia. Los objetivos básicos son: procurar información a las personas mayores sobre los aspectos que determinan un envejecimiento activo y saludable; dotar a las personas mayores de las habilidades necesarias para prevenir las situaciones de dependencia y lograr un envejecimiento satisfactorio; promover la autonomía de las personas mayores con discapacidad; facilitar a los profesionales de la salud y los servicios sociales herramientas para conseguir un envejecimiento saludable y prevenir la dependencia, y, finalmente, generar un cambio de actitudes y un pensamiento positivo ante el envejecimiento (AU)


The present article describes a research-action project based on qualitative methodology, which aims to promote active and successful ageing and prevent dependency. The main goals of the project are: to offer information to older adults on the factors leading to active and successful ageing; to provide elderly people with the skills needed to prevent dependency and achieve satisfactory ageing; to promote autonomy in elderly people with disabilities; to supply health and social services professionals with tools to achieve successful ageing and prevent dependency; and to change attitudes and generate positive thinking about ageing (AU)


Assuntos
Humanos , Idoso , Idoso de 80 Anos ou mais , Educação em Saúde/organização & administração , Programas Gente Saudável/organização & administração , Envelhecimento , Transtornos das Habilidades Motoras/prevenção & controle , Doença Crônica/prevenção & controle , Atitude Frente a Saúde , Acesso à Informação , Percepção Social , Materiais de Ensino/provisão & distribuição
7.
Ansiedad estrés ; 11(2/3): 215-231, dic. 2005. ilus, tab
Artigo em Es | IBECS | ID: ibc-042360

RESUMO

En este estudio se pone a prueba el modelo espiral 'hacia arriba' y 'hacia abajo' de las creencias de eficacia en muestras de estudiantes universitarios (N=353) de los cuales 203 son españoles y 150 son belgas. Modelos de ecuaciones estructurales muestran apoyo empírico al modelo 'hacia abajo' del burnout como 'crisis de eficacia' y al modelo 'hacia arriba' de la eficacia como 'motor del engagement'. También los resultados muestran apoyo empírico a la mediación de la eficacia percibida entre el éxito pasado y los niveles actuales de burnout y de engagement; que a su vez predicen la autoeficacia en el éxito académico. Por otra parte, aunque distintos ANOVAs muestran diferencias significativas en función del país en algunas variables estudiadas, los análisis multigrupo con modelos de ecuaciones estructurales muestran la invarianza del modelo a través de ambas muestras. Finalmente, análisis estructurales en profundidad con la muestra de estudiantes belgas pone de manifiesto la necesidad de considerar también las creencias de 'ineficacia' en futuros estudios


In this study the Downward and Upward Spiral Model of efficacy beliefs is tested in two samples of Spanish (N=203) and Belgian (N=150) university students respectively. Results from Structural Equations Modeling showed empirical support for the Downward Spiral Model of burnout , which is considered as a "crisis of efficacy", and for the Upward Spiral Model of engagement, which as a 'boost of efficacy '. Results also show empirical support to the mediating role that efficacy beliefs play in the relationship between past success and burnout/engagement on the one hand, and self-efficacy beliefs about academic success on the other hand. Moreover, using multigroup analysis it was shown that the research model was invariant of across both samples


Assuntos
Masculino , Feminino , Adulto , Humanos , Esgotamento Profissional/psicologia , Avaliação Educacional/métodos , Estresse Psicológico/psicologia , Estudantes/psicologia , Confiança/psicologia , Competência Profissional , Eficiência , Aprendizagem
8.
Cell Calcium ; 32(4): 193-200, 2002 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-12379179

RESUMO

Striated muscle represents one of the best models for studies on Ca(2+) signalling. However, although much is known on the localisation and molecular interactions of the ryanodine receptors (RyRs), far less is known on the localisation and on the molecular interactions of the inositol trisphosphate receptors (InsP(3)Rs) in striated muscle cells. Recently, members of the Homer protein family have been shown to cluster type 1 metabotropic glutamate receptors (mGluR1) in the plasma membrane and to interact with InsP(3)R in the endoplasmic reticulum of neurons. Thus, these scaffolding proteins are good candidates for organising plasma membrane receptors and intracellular effector proteins in signalosomes involved in intracellular Ca(2+) signalling. Homer proteins are also expressed in skeletal muscle, and the type 1 ryanodine receptor (RyR1) contains a specific Homer-binding motif. We report here on the relative sub-cellular localisation of InsP(3)Rs and Homer proteins in skeletal muscle cells with respect to the localisation of RyRs. Immunofluorescence analysis showed that both Homer and InsP(3)R proteins present a staining pattern indicative of a localisation at the Z-line, clearly distinct from that of RyR1. Consistent herewith, in sub-cellular fractionation experiments, Homer proteins and InsP(3)R were both found in the fractions enriched in longitudinal sarcoplasmic reticulum (LSR) but not in fractions of terminal cisternae that are enriched in RyRs. Thus, in skeletal muscle, Homer proteins may play a role in the organisation of a second Ca(2+) signalling compartment containing the InsP(3)R, but are apparently not involved in the organisation of RyRs at triads.


Assuntos
Canais de Cálcio/metabolismo , Proteínas de Transporte/metabolismo , Músculo Esquelético/metabolismo , Neuropeptídeos/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Retículo Sarcoplasmático/metabolismo , Anticorpos/imunologia , Cálcio/metabolismo , Canais de Cálcio/imunologia , Proteínas de Transporte/imunologia , Imunofluorescência , Proteínas de Arcabouço Homer , Receptores de Inositol 1,4,5-Trifosfato , Fibras Musculares Esqueléticas/metabolismo , Neuropeptídeos/imunologia , Receptores Citoplasmáticos e Nucleares/imunologia , Canal de Liberação de Cálcio do Receptor de Rianodina/imunologia , Canal de Liberação de Cálcio do Receptor de Rianodina/metabolismo , Transdução de Sinais/fisiologia
9.
J Biol Chem ; 276(14): 11189-98, 2001 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-11134006

RESUMO

Subcellular targeting of the components of the cAMP-dependent pathway is thought to be essential for intracellular signaling. Here we have identified a novel protein, named myomegalin, that interacts with the cyclic nucleotide phosphodiesterase PDE4D, thereby targeting it to particulate structures. Myomegalin is a large 2,324-amino acid protein mostly composed of alpha-helical and coiled-coil structures, with domains shared with microtubule-associated proteins, and a leucine zipper identical to that found in the Drosophila centrosomin. Transcripts of 7.5-8 kilobases were present in most tissues, whereas a short mRNA of 2.4 kilobases was detected only in rat testis. A third splicing variant was expressed predominantly in rat heart. Antibodies against the deduced sequence recognized particulate myomegalin proteins of 62 kDa in testis and 230-250 kDa in heart and skeletal muscle. Immunocytochemistry and transfection studies demonstrate colocalization of PDE4D and myomegalin in the Golgi/centrosomal area of cultured cells, and in sarcomeric structures of skeletal muscle. Myomegalin expressed in COS-7 cells coimmunoprecipitated with PDE4D3 and sequestered it to particulate structures. These findings indicate that myomegalin is a novel protein that functions as an anchor to localize components of the cAMP-dependent pathway to the Golgi/centrosomal region of the cell.


Assuntos
3',5'-AMP Cíclico Fosfodiesterases/metabolismo , Centrossomo/metabolismo , Complexo de Golgi/metabolismo , Proteínas/metabolismo , Sequência de Aminoácidos , Animais , Drosophila , Imuno-Histoquímica , Proteínas Associadas aos Microtúbulos/análise , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Dados de Sequência Molecular , Especificidade de Órgãos , Ligação Proteica , Proteínas/análise , Proteínas/genética , Ratos , Saccharomyces cerevisiae , Análise de Sequência
10.
Lab Invest ; 79(9): 1127-35, 1999 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-10496531

RESUMO

The possibility of diagnosing neoplastic testis pathologies by studying immature germ cells released from the seminiferous epithelium and present in the semen has been reported. It has been suggested that carcinoma in situ (CIS) of the testis and testis tumor may be identified by studying specific surface antigenic determinants or ploidy of chromosome 1 of malignant germ cells recovered from the semen. A noninvasive diagnostic approach of this type would be of great interest if we consider that CIS is supposed to precede the development of testicular germ cell tumors and that the frequency of that preneoplastic condition is increased in specific andrologic pathologies. To evaluate the reliability of this diagnostic approach, we have quantified the presence of immature hyperdiploid germ cells in the seminal fluid of normal healthy subjects, of infertile oligozoospermic patients affected by maldescended testis or left vancocele, and of patients suffering from CIS or testis tumor. Cell ploidy was evaluated on seminal cell fractions highly enriched in immature germ cells, by means of in situ hybridization with a DNA-probe specific for chromosome 1. Our observations indicate that chromosome 1 hyperdiploidy is not necessarily a predictive parameter of testis tumoral pathologies. The percentage of hyperdiploid immature seminal germ cells is in fact increased in nontumoral pathologies associated with infertility, such as cryptorchidism and left varicocele.


Assuntos
Diploide , Ejaculação/fisiologia , Espermatozoides/patologia , Neoplasias Testiculares/diagnóstico , Adulto , Estudos de Casos e Controles , Senescência Celular/fisiologia , Humanos , Hibridização In Situ , Masculino , Oligospermia/diagnóstico , Oligospermia/genética , Valor Preditivo dos Testes , Neoplasias Testiculares/genética
11.
Endocrinology ; 140(5): 2297-306, 1999 May.
Artigo em Inglês | MEDLINE | ID: mdl-10218983

RESUMO

The type 4 cAMP-specific phosphodiesterases (PDE4) are a family of closely related enzymes with similar catalytic domains and divergent amino- and carboxyl-terminus domains. Multiple PDE proteins with heterogeneous amino termini are derived from each gene. To understand the significance of this heterogeneity, the expression and localization of variants derived from PDE4A and PDE4D genes was investigated during spermatogenesis in the rat. RNase protection analysis with mRNA for testes at different ages of development showed that two transcripts (PDE4D1 and PDE4D2) are expressed at day 10 and 15 of age and become undetectable thereafter. An additional PDE4D transcript appears at day 30 and increased during testid maturation. This latter transcript codes for a long variant of the PDE4D gene and is expressed in germ cells as demonstrated by RNase protection with RNA from isolated pachytene spermatocytes and round spermatids. The presence of a corresponding PDE4D protein with a molecular mass of 98 kDa was established by immunoprecipitation and Western blot analysis with antibodies specific for PDE4D and by immunoaffinity chromatography purification of the 98 kDa variant from isolated germ cells. PDE4A transcripts were also expressed in pachytene spermatocytes and round spermatids. Two polypeptides encoded by these PDE4A transcripts were expressed in pachytene spermatocytes, reached a maximum in round spermatids, and declined thereafter. Immunofluorescence analysis demonstrated a localization of the PDE4D protein in the manchette and in a periacrosomal region of the developing spermatid, a localization confirmed by immunogold electron microscopy. Conversely, the PDE4A was mostly soluble in the cytoplasm of round spermatids. These data demonstrate that PDE4D and PDE4A variants are expressed at different stages and localized in distinct subcellular structures of developing spermatids. Different properties of the mRNAs derived from the two genes and localization signals are responsible for the temporal and spatial expression of the different PDE4 isoenzymes.


Assuntos
3',5'-AMP Cíclico Fosfodiesterases/genética , Expressão Gênica , Isoenzimas/genética , Espermatogênese , Testículo/enzimologia , Animais , Imunofluorescência , Hormônio Foliculoestimulante/farmacologia , Variação Genética , Masculino , Microscopia Imunoeletrônica , Splicing de RNA , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Túbulos Seminíferos/enzimologia , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/enzimologia , Espermatozoides/enzimologia , Testículo/crescimento & desenvolvimento
12.
Endocrinology ; 139(12): 4839-48, 1998 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9832420

RESUMO

Apoptosis is a physiological process by which multicellular organisms eliminate unwanted cells. Death factors such as Fas ligand induce apoptosis by triggering a series of intracellular protein-protein interactions mediated by defined motifs found in the signaling molecules. One of these motifs is the death effector domain (DED), a stretch of about 80 amino acids that is shared by adaptors, regulators, and executors of the death factor pathway. We have identified the human and rat complementary DNAs encoding a novel protein termed DEFT (Death EFfector domain-containing Testicular molecule). The N-terminus of DEFT shows a high degree of homology to the DEDs found in FADD (an adaptor molecule) as well as procaspase-8/FLICE and procaspase-10/Mch4 (executors of the death program). Northern blot hybridization experiments have shown that the DEFT messenger RNA (mRNA) is expressed in a variety of human and rat tissues, with particularly abundant expression in the testis. In situ hybridization analysis further indicated the expression of DEFT mRNA in meiotic male germ cells. In a model of germ cell apoptosis induction, an increase in testis DEFT mRNA was found in immature rats after 2 days of treatment with a GnRH antagonist. Unlike FADD and procaspase-8/FLICE, overexpression of DEFT did not induce apoptosis in Chinese hamster ovary cells. Although cotransfection studies indicated that DEFT is incapable of modulating apoptosis effected by FADD and procaspase-8/FLICE, interactions between DEFT and uncharacterized DED-containing molecules in the testis remain to be studied in the future. In conclusion, we have identified a novel DED-containing protein with high expression in testis germ cells. This protein may be important in the regulation of death factor-induced apoptosis in the testis and other tissues.


Assuntos
Proteínas de Ligação a DNA , Células Germinativas/metabolismo , Proteínas/genética , Testículo/metabolismo , Sequência de Aminoácidos , Animais , Apoptose/fisiologia , Sequência de Bases , Células CHO , Clonagem Molecular , Cricetinae , Proteínas Adaptadoras de Sinalização de Receptores de Domínio de Morte , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Dados de Sequência Molecular , Proteínas/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Homologia de Sequência de Aminoácidos , Testículo/citologia , Testículo/fisiologia
13.
Endocrinology ; 139(5): 2588-99, 1998 May.
Artigo em Inglês | MEDLINE | ID: mdl-9564875

RESUMO

To identify the adenylyl cyclase (AC) genes expressed in mammalian germ cells, RT-PCR of testis and germ cell RNA was performed using degenerated primers based on the homologous region of the AC catalytic domain. This strategy yielded high-frequency amplification of a complementary DNA (cDNA) identical to type III AC (ACIII), a form previously identified as the major adenylyl cyclase expressed in the olfactory system. Ribonuclease protection studies confirmed that ACIII transcripts are present in germ cells, appear during the meiotic prophase, and accumulate during spermiogenesis. A Northern blot analysis performed on total testis RNA demonstrated the presence of a predominant transcript of 7.5 kb, suggesting that the ACIII expressed in germ cells may derive from a splicing variant different from the 4.5 kb transcripts expressed in somatic cells. To determine whether these RNAs are translated into a protein, Western blot analysis was performed using an antibody specific for the carboxyl terminus of ACIII. An immunoreactive protein of 170 kDa was detected in extracts from total testis and from germ cells. Immunofluorescence localization of this protein in the seminiferous tubules showed that ACIII was predominantly expressed in postmeiotic germ cells from round spermatids in the cap phase to maturing elongating spermatids. The ACIII antigen was located mostly on the acrosomal membrane rather than on the plasma membrane of developing spermatids. The spatial and temporal expression of ACIII in germ cells indicates a role of this AC in the acrosome formation. Together with the observation that members of the olfactory receptor family and an olfactory phosphodiesterase are expressed in spermatids, these findings suggest that a signal transduction system used in olfaction is also used during gamete development.


Assuntos
Adenilil Ciclases/genética , Expressão Gênica , Espermatogênese , Espermatozoides/enzimologia , Adenilil Ciclases/química , Sequência de Aminoácidos , Animais , Northern Blotting , Western Blotting , Masculino , Dados de Sequência Molecular , Mucosa Olfatória/enzimologia , Reação em Cadeia da Polimerase , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Homologia de Sequência , Espermátides/enzimologia , Testículo/enzimologia
14.
Methods ; 14(1): 55-64, 1998 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-9500858

RESUMO

With the cloning of cDNAs coding for the different phosphodiesterase 4 (PDE4) isoenzymes present in mammals, homogeneous preparations of these forms have become readily available. This strategy has greatly facilitated the understanding of the properties of the myriad of isoforms derived from the four PDE4 genes found in mammals, and has opened a new avenue to develop inhibitors with a different degree of selectivity for each isoform. Here we describe the strategies and methods used to express PDE4 in bacterial, yeast, insect, and mammalian cell heterologous systems, and review the advantages and disadvantages of each of these expression strategies. In addition, procedures to purify the recombinant proteins are described. The recently developed purification of a PDE4 by immunoaffinity chromatography provides a rapid and efficient method to prepare large quantities of PDE4. This method should be very useful for structural and kinetic studies on the PDE4D isoforms.


Assuntos
3',5'-AMP Cíclico Fosfodiesterases/genética , Expressão Gênica/genética , Pirrolidinonas/farmacologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , 3',5'-AMP Cíclico Fosfodiesterases/isolamento & purificação , Animais , Baculoviridae/genética , Linhagem Celular , Cromatografia de Afinidade , Cromatografia Líquida de Alta Pressão , Nucleotídeo Cíclico Fosfodiesterase do Tipo 3 , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4 , Escherichia coli/enzimologia , Humanos , Ratos , Rolipram , Saccharomyces cerevisiae/genética , Spodoptera/genética
15.
Biol Reprod ; 58(2): 371-8, 1998 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9475391

RESUMO

The distribution of alpha6beta1 integrins at the level of cell-to-cell contacts within the rat seminiferous epithelium was investigated. Double fluorescence experiments using phalloidin staining of actin filaments and anti-integrin subunit antibodies showed that the receptor belongs to the Sertoli cell lateral domains engaged in the characteristic junctional structures known as ectoplasmic specializations (ES), at the level both of inter-Sertoli junctions and of the contacts between Sertoli cells and elongating spermatids. In the seminiferous epithelium of aspermatogenic testes, obtained through X-irradiation in utero (Sertoli-cell-only testes), at the level of inter-Sertoli junctions both ES and alpha6beta1 integrins are present. In order to study the dependence of alpha6beta1 receptors and ES formation upon FSH stimulation during development, 9-day-old testes were grown in organ culture in basal as well as FSH-supplemented conditions. FSH stimulation, which is necessary for the progression of spermatogenesis to early meiotic stages, appears to be required for the development of inter-Sertoli junctional structures containing ES and alpha6beta1 integrins. These observations indicate that the receptor belongs to the inter-Sertoli junctional machinery and that its expression at that level is not dependent on active spermatogenesis but requires FSH stimulation.


Assuntos
Antígenos CD59/biossíntese , Hormônio Foliculoestimulante/fisiologia , Integrinas/biossíntese , Epitélio Seminífero/metabolismo , Epitélio Seminífero/fisiologia , Células de Sertoli/fisiologia , Espermatogênese/fisiologia , Animais , Antígenos CD59/genética , Comunicação Celular/fisiologia , Corantes Fluorescentes , Hormônio Foliculoestimulante/biossíntese , Integrina alfa6beta1 , Integrinas/genética , Laminina/biossíntese , Masculino , Microscopia Eletrônica , Microscopia de Fluorescência , Técnicas de Cultura de Órgãos , Ratos , Ratos Wistar , Epitélio Seminífero/citologia , Células de Sertoli/ultraestrutura , Testículo/citologia , Testículo/crescimento & desenvolvimento
16.
Poult Sci ; 76(10): 1405-17, 1997 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9316117

RESUMO

Three experiments were conducted to determine the effects of supplementing practical diets of male turkeys with dl-alpha-tocopheryl acetate (TA). In Experiment 1, a factorial arrangement of dietary treatments [0, 12, 50, 150, and 300 IU TA/kg with 0 or 300 mg ascorbic acid (AA)/kg] was used. These 10 treatments were fed to poults from 1 to 41 d of age. From 41 to 118 d of age, the AA treatments were discontinued, and the 300 IU TA treatment groups were changed to 12 IU TA/kg. Neither TA nor AA treatments affected 41-d BW, feed to gain ratio (FE), or livability. No effects of dietary TA concentrations on turkey performance were observed through 118 d of age alpha-Tocopherol (TOC) concentrations of plasmas and livers were increased by increments of dietary TA, with substantial liver storage when toms were fed 150 IU TA/kg from 1 to 118 d. Supplementing diets with 0, 25, 50, 75, or 100 IU TA/ kg in Experiments 2 and 3 had no effect on performance of toms through 119 and 105 d, respectively. alpha-Tocopherol concentrations of plasma and red blood cells (RBC) increased linearly with increments of dietary TA. The same was true for livers in Experiment 2. Susceptibility of RBC to hemolysis induced by 400 microM t-butyl hydroperoxide (TBH) in Experiment 2 decreased with increasing dietary TA, and these decreases corresponded to increases in TOC concentration of RBC. However, the relationships between hemolysis and dietary TA or RBC TOC were inconsistent in Experiment 3 and varied according to concentration of TBH (200, 300, or 400 microM) and age of the toms. At 105 d of age, RBC of toms fed no supplemental TA were resistant to hemolysis, irrespective of dietary TA and TBH concentration. In Experiment 3, there were no indications of dietary TA effects on plasma peroxide concentration or activity of plasma creatine kinase. A positive relationship between dietary TA and blastogenic responses of blood lymphocytes was observed with concanavalin A when toms were at 44 d but not at 23 or 86 d of age. The overall data indicate that corn-soybean meal diets containing from 6 to 20 IU TOC/kg, but no supplemental TA supported satisfactory performance and well-being of male turkeys from 1 d of age to market ages when the turkeys were free of disease, as was true in the research reported here.


Assuntos
Dieta/veterinária , Glycine max/normas , Perus/crescimento & desenvolvimento , Perus/fisiologia , Vitamina E/farmacologia , Zea mays/normas , Envelhecimento/fisiologia , Análise de Variância , Animais , Peso Corporal/fisiologia , Concanavalina A/farmacologia , Suplementos Nutricionais , Relação Dose-Resposta a Droga , Contagem de Eritrócitos , Lipopolissacarídeos/farmacologia , Fígado/química , Ativação Linfocitária/fisiologia , Masculino , Peróxidos/farmacologia , Lectinas de Plantas , Análise de Regressão , Perus/sangue , Vitamina E/administração & dosagem , Vitamina E/análise , terc-Butil Hidroperóxido
17.
Poult Sci ; 75(11): 1393-403, 1996 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8933593

RESUMO

An experiment was conducted to compare the efficacy of two dietary sources and an injectable form of vitamin E (VE) to improve the VE status of poults. Six of the treatments consisted of a factorial arrangement of three concentrations and two sources of dietary VE. Turkeys in these treatments received 12, 80, or 150 IU of either dl-alpha-tocopheryl acetate or d-alpha-tocopherol (d-alpha-TOC)/kg of diet. The seventh treatment consisted of a single subcutaneous injection of d-alpha-TOC at 1 d of age. Poults in this treatment were subcutaneously injected in the dorsal area of the neck with 25 IU of d-alpha-TOC, this amount being approximately equivalent to the amount poults would consume if their diet was supplemented with 150 IU of VE/kg during their 1st wk of life. Concentration, source, or route of VE administration did not affect growth parameters, plasma creatine kinase, plasma triglycerides, or liver lipid peroxidation as measured by the thiobarbituric acid reactive substances assay (TBARS). Plasma, red blood cells (RBC), and liver alpha-TOC decreased from hatching to 14 d of age in poults fed either source of VE. The use of 80 or 150 IU of dietary VE (either source) reduced (P < 0.05) the extent of depletion of alpha-TOC at all ages and also reduced the susceptibility of RBC to hemolysis. There was no effect of source of dietary VE on concentration of alpha-TOC in plasma, RBC, or liver, or on RBC hemolysis. Subcutaneous injection of 25 IU of d-alpha-TOC at Day 1 increased (P < 0.05) alpha-TOC concentration until 7 d of age. Also, d-alpha-TOC injection reduced (P < 0.05) RBC susceptibility to hemolysis through 21 d of age. Data showed that one single subcutaneous injection of 25 IU of d-alpha-TOC at 1 d of age was as effective as 80 IU or more of dietary VE through 21 d to improve the alpha-TOC status of poults.


Assuntos
Dieta/veterinária , Perus/sangue , Vitamina E/administração & dosagem , Vitamina E/farmacologia , Administração Oral , Animais , Peso Corporal/fisiologia , Creatina Quinase/fisiologia , Dieta/normas , Relação Dose-Resposta a Droga , Eritrócitos/química , Injeções Subcutâneas/veterinária , Masculino , Distribuição Aleatória , Glycine max/normas , Fatores de Tempo , Triglicerídeos/sangue , Perus/fisiologia , Vitamina E/análise , Zea mays/normas
18.
Poult Sci ; 74(9): 1470-83, 1995 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-7501592

RESUMO

An experiment was conducted to determine the effect of two early nutrient restriction programs on performance, selected characteristics of the gastrointestinal tract (GIT), and activities of digestive enzymes of broiler chickens. Three hundred and sixty male broiler (Ross x Ross) chicks kept in floor pens were assigned to three groups. The control group (C) was given ad libitum access to feed from 1 to 48 d of age. Another group was restricted from 11 to 14 d (R4) of age to an energy intake of .74 x BW.67 kcal ME/d, and a third group was restricted from 7 to 14 d (R7) of age to an energy intake of 1.5 x BW.67 kcal ME/d. Then, both restricted groups were given ad libitum access to feed through 48 d. Body weight and feed intake were determined weekly and selected carcass characteristics were measured at 48 d of age. Broilers also were sampled at 7, 14, 21, and 42 d of age to obtain data on components of the GIT (proventriculus, gizzard, pancreas, and small intestine) and activities of selected digestive enzymes. Feed-restricted groups were lighter in body weight (P < .01) at 14 and 48 d of age than the C group but were superior in overall feed efficiency. No treatment effects were observed for percentage yields of breast meat and abdominal fat pad. Absolute weights of GIT components were significantly reduced at 14 d of age by feed restriction. However, GIT components increased in weight more quickly after refeeding than did the whole body. Restricted groups had reduced (P < .01) specific activities of jejunal alkaline phosphatase and pancreatic trypsin, amylase, and lipase as compared with the C group at 14 d of age but not at 21 and 42 d of age. Relative activities for jejunal maltase and sucrase were greater (P < .01) at 21 d of age in the R4 and R7 groups than in the C group. The present data show that feed restriction results in transient changes in organs and activities of digestive enzymes, suggesting a functional adaptation to feed restriction.


Assuntos
Peso Corporal/fisiologia , Galinhas/fisiologia , Sistema Digestório/anatomia & histologia , Privação de Alimentos/fisiologia , Jejuno/enzimologia , Pâncreas/enzimologia , Fatores Etários , Animais , Galinhas/metabolismo , Masculino , Tamanho do Órgão
19.
Poult Sci ; 74(6): 983-97, 1995 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-7644428

RESUMO

Two experiments were conducted to document the effects of an early immunologic stress and changes in dietary ME(n) on growth and nutrient utilization of newly hatched turkeys. Treatments in both experiments consisted of a complete factorial arrangement of two types of injection and four isonitrogenous diets. Turkeys were injected i.p. with saline (SAL) or a solution of lipopolysaccharide (LPS) (100 micrograms LPS/mL SAL) at 1, 3, and 5 d of age. In Experiment 1, two diets were formulated to contain 2,800 kcal ME(n)/kg. One was a corn-soybean meal-based diet (CSBM) and the other contained 8% Solkafloc (SKF). A third diet (3,100 kcal ME(n)/kg) was formulated by substituting 8% sucrose (SUC) for the 8% SKF. The fourth diet included in Experiment 1 was formulated to contain 3,700 kcal ME(n)/kg. The CSBM and SUC diets were also included in Experiment 2. Two additional diets tested in Experiment 2 were the CSBM diet containing 74.5 mg ibuprofen/kg (IBU) and a corn-soybean meal-based diet with a ME(n) value of 3,100 kcal/kg (CS31). Injection with LPS reduced (P < .05) BW of turkeys throughout Experiment 1 and until 9 d of age in Experiment 2, as compared with injection with SAL, irrespective of dietary treatment. The reduction in BW was mainly due to a decrease in feed intake (FI) (P < .05). Turkeys fed diets with 3,100 kcal ME(n)/kg were heavier (P < .05) than those fed diets with 2,800 kcal ME(n)/kg, irrespective of injection. Inclusion of ibuprofen to the CSBM diet from 1 to 14 d improved (P < .05) BW and feed efficiency (P < .01) of turkeys at 14 d of age, compared with turkeys fed the CSBM diet. Determined ME(n) was not affected by LPS injection. Adverse effects of LPS injection on growth of turkey poults were mainly the consequence of a reduced FI and not of altered nutrient utilization. These effects were not fully alleviated by feeding a diet with 3,100 kcal ME(n)/kg.


Assuntos
Ração Animal , Dieta/veterinária , Metabolismo Energético , Crescimento/fisiologia , Lipopolissacarídeos/farmacologia , Perus/fisiologia , Animais , Ingestão de Energia , Escherichia coli , Crescimento/imunologia , Lipopolissacarídeos/imunologia , Masculino , Valores de Referência , Glycine max , Zea mays
20.
Biol Reprod ; 52(1): 79-87, 1995 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-7711187

RESUMO

With the aim of investigating the presence and role of integrin receptors in cell-to-cell interactions during spermatogenesis, we have immunolocalized alpha 6A integrin chain in the rat seminiferous epithelium. In both prepubertal and adult seminiferous epithelium, the antigen was found to be restricted to definite sites of intercellular contact, following a stage-specific distribution almost invariably identical to that known for beta 1 chain. In the adult seminiferous epithelium, positivity for both antigens was found exclusively around the profiles of elongating and maturing spermatids and, in most but not all stages, at a characteristic suprabasal position. In the prepubertal rat, the antigen is localized along a very regular suprabasal line of intercellular contacts. In immunoprecipitation experiments on both seminiferous epithelium explants and Sertoli cell cultures from 3-wk-old rats, anti-alpha 6A antibody coprecipitates a polypeptide of 118 kDa, presumably corresponding to beta 1 chain. These data strongly suggest that the integrin heterodimer alpha 6A beta 1 is expressed at sites of intercellular contact in the rat seminiferous epithelium. The stage-specific and restricted pattern observed by immunofluorescence suggests that this integrin is involved in regulatory interactions between Sertoli cells and germ cells during spermatogenesis.


Assuntos
Integrinas/metabolismo , Epitélio Seminífero/metabolismo , Animais , Sítios de Ligação , Adesão Celular/fisiologia , Comunicação Celular/fisiologia , Células Cultivadas , Imuno-Histoquímica , Integrina alfa6beta1 , Masculino , Ratos , Ratos Wistar , Receptores de Laminina/metabolismo , Epitélio Seminífero/citologia , Células de Sertoli/metabolismo , Maturidade Sexual/fisiologia
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