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1.
Cell Death Dis ; 4: e586, 2013 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-23579273

RESUMO

Neuroblastoma is an embryonal malignancy of the sympathetic nervous system. Spontaneous regression and differentiation of neuroblastoma is observed in a subset of patients, and has been suggested to represent delayed activation of physiologic molecular programs of fetal neuroblasts. Homeobox genes constitute an important family of transcription factors, which play a fundamental role in morphogenesis and cell differentiation during embryogenesis. In this study, we demonstrate that expression of the majority of the human HOX class I homeobox genes is significantly associated with clinical covariates in neuroblastoma using microarray expression data of 649 primary tumors. Moreover, a HOX gene expression-based classifier predicted neuroblastoma patient outcome independently of age, stage and MYCN amplification status. Among all HOX genes, HOXC9 expression was most prominently associated with favorable prognostic markers. Most notably, elevated HOXC9 expression was significantly associated with spontaneous regression in infant neuroblastoma. Re-expression of HOXC9 in three neuroblastoma cell lines led to a significant reduction in cell viability, and abrogated tumor growth almost completely in neuroblastoma xenografts. Neuroblastoma growth arrest was related to the induction of programmed cell death, as indicated by an increase in the sub-G1 fraction and translocation of phosphatidylserine to the outer membrane. Programmed cell death was associated with the release of cytochrome c from the mitochondria into the cytosol and activation of the intrinsic cascade of caspases, indicating that HOXC9 re-expression triggers the intrinsic apoptotic pathway. Collectively, our results show a strong prognostic impact of HOX gene expression in neuroblastoma, and may point towards a role of Hox-C9 in neuroblastoma spontaneous regression.


Assuntos
Regulação Neoplásica da Expressão Gênica , Proteínas de Homeodomínio/genética , Regressão Neoplásica Espontânea/genética , Neoplasias do Sistema Nervoso/genética , Neuroblastoma/genética , Apoptose/genética , Caspases/genética , Caspases/metabolismo , Diferenciação Celular , Linhagem Celular Tumoral , Pré-Escolar , Citocromos c/metabolismo , Proteínas de Homeodomínio/metabolismo , Humanos , Lactente , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Proteína Proto-Oncogênica N-Myc , Estadiamento de Neoplasias , Neoplasias do Sistema Nervoso/metabolismo , Neoplasias do Sistema Nervoso/mortalidade , Neoplasias do Sistema Nervoso/patologia , Neuroblastoma/metabolismo , Neuroblastoma/mortalidade , Neuroblastoma/patologia , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Proteínas Oncogênicas/genética , Proteínas Oncogênicas/metabolismo , Prognóstico , Transdução de Sinais , Análise de Sobrevida , Ensaios Antitumorais Modelo de Xenoenxerto
2.
Leukemia ; 23(5): 892-9, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19158832

RESUMO

We found that composition of cell subsets within the CD34+ cell population is markedly altered in chronic phase (CP) chronic myeloid leukemia (CML). Specifically, proportions and absolute cell counts of common myeloid progenitors (CMP) and megakaryocyte-erythrocyte progenitors (MEP) are significantly greater in comparison to normal bone marrow whereas absolute numbers of hematopoietic stem cells (HSC) are equal. To understand the basis for this, we performed gene expression profiling (Affymetrix HU-133A 2.0) of the distinct CD34+ cell subsets from six patients with CP CML and five healthy donors. Euclidean distance analysis revealed a remarkable transcriptional similarity between the CML patients' HSC and normal progenitors, especially CMP. CP CML HSC were transcriptionally more similar to their progeny than normal HSC to theirs, suggesting a more mature phenotype. Hence, the greatest differences between CP CML patients and normal donors were apparent in HSC including downregulation of genes encoding adhesion molecules, transcription factors, regulators of stem-cell fate and inhibitors of cell proliferation in CP CML. Impaired adhesive and migratory capacities were functionally corroborated by fibronectin detachment analysis and transwell assays, respectively. Based on our findings we propose a loss of quiescence of the CML HSC on detachment from the niche leading to expansion of myeloid progenitors.


Assuntos
Regulação Leucêmica da Expressão Gênica/genética , Células-Tronco Hematopoéticas/patologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Células Progenitoras Mieloides/patologia , Adesão Celular , Diferenciação Celular , Movimento Celular , Proliferação de Células , Imunofluorescência , Perfilação da Expressão Gênica , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Células-Tronco Neoplásicas , Análise de Sequência com Séries de Oligonucleotídeos , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa
3.
J Microsc ; 223(Pt 2): 133-9, 2006 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16911073

RESUMO

The appearance of lanthanum in liver cells as a result of the injection of lanthanum chloride into rats is investigated by advanced transmission electron microscopy techniques, including electron energy loss spectroscopy and high-resolution transmission electron microscopy. It is demonstrated that the lysosomes contain large amounts of lanthanum appearing in a granular form with particle dimensions between 5 and 25 nm, whereas no lanthanum could be detected in other surrounding cellular components.


Assuntos
Hepatócitos/química , Lantânio/análise , Microscopia Eletrônica de Transmissão , Espectrometria por Raios X , Espectroscopia de Perda de Energia de Elétrons , Animais , Lisossomos/química , Modelos Animais , Ratos
4.
Rev Neurol ; 36(11): 1030-4, 2003.
Artigo em Espanhol | MEDLINE | ID: mdl-12808498

RESUMO

INTRODUCTION: Zellweger syndrome, or cerebrohepatorenal syndrome, is the most serious form of the peroxisomal diseases. Clinically, it is characterised by the association between craniofacial dysmorphia and neurological disorders, together with the involvement of other organs. To perform a diagnosis it is advisable to follow a procedural protocol that begins with the quantification of very long chain fatty acids in an assortment of samples (serum, fibroblasts and mononucleate cells), plasmalogens, branched chain fatty acids in serum (phytanic and pristanic acids), polyunsaturated acids and bile salts. Studies conducted with neuroimaging, renal echography, skeletal X rays and biopsy samples of different tissues will provide us with information about the involvement of different organs. CASE REPORTS: The first case we report is that of a male who, from birth, presented a distinctive phenotype with very large fontanelles, important hypotonia, epileptic seizures and acute organic disorders that led to death at the age of seven weeks. The second case involved a new born male suffering from prenatally diagnosed heart disease and craniofacial dysmorphia with hypotonia. CONCLUSIONS: Given the scarce survival rate of patients, early diagnosis of this type of disease is crucial and, although complex, a study must be conducted so as to be able to provide genetic counselling.


Assuntos
Síndrome de Zellweger/diagnóstico , Síndrome de Zellweger/fisiopatologia , Córtex Cerebral/patologia , Cistos/patologia , Evolução Fatal , Ácidos Graxos/sangue , Humanos , Lactente , Recém-Nascido , Masculino , Fenótipo
5.
Rev. neurol. (Ed. impr.) ; 36(11): 1030-1034, 1 jun., 2003.
Artigo em Es | IBECS | ID: ibc-27656

RESUMO

Introducción. El síndrome de Zellweger o síndrome cerebro hepatorrenales la forma más grave de las enfermedades peroxisomales. Clínicamente, se caracteriza por la asociación de dismorfia craneofacial y alteraciones neurológicas, junto con la afectación de otros órganos. Para realizar el diagnóstico es conveniente recurrir a un protocolo de actuación que comienza con la cuantificación de los ácidos grasos de cadena muy larga en diversas muestras (suero,fibroblastos y células mononucleadas), plasmalógenos, ácidos ramificados en el suero (ácidos fitánico y pristánico), ácidos poliinsaturados y sales biliares; continúa con los estudios de neuroimagen, ecografía renal, radiografías de esqueleto, y concluye con la toma de muestras mediante biopsia de diversos tejidos, que nos aportará información acerca de la afectación de distintos órganos. Casos clínicos. Presentamos un primer caso de un varón que ya desde el nacimiento presenta un fenotipo peculiar, con fontanelas muy amplias, hipotonía grave, crisis epilépticas y agudas alteraciones orgánicas que condujeron al fallecimiento a las siete semanas de vida. El segundo caso corresponde a un recién nacido varón afectado de una cardiopatía diagnosticada prenatalmente y dismorfia craneofacial con hipotonía. Conclusiones.El diagnóstico precoz de este tipo de enfermedades es crucial, dada la escasa supervivencia de los pacientes y la necesidad de realizar un estudio que, a pesar de resultar complejo, es necesario para poder ofrecer un consejo genético (AU)


Assuntos
Masculino , Recém-Nascido , Lactente , Humanos , Síndrome de Zellweger , Evolução Fatal , Fenótipo , Córtex Cerebral , Cistos , Ácidos Graxos
6.
Cells Tissues Organs ; 173(1): 46-53, 2003.
Artigo em Inglês | MEDLINE | ID: mdl-12566626

RESUMO

The endometrial surface morphology of 38 dogs during different stages of the estrous cycle was investigated with scanning electron microscopy. The cell surface altered from convex in proestrus and estrus to very variable in early metestrus, flattened in late metestrus and became completely plane in anestrus. Microvilli were numerous and long in proestrus and in estrus, became short and variable in number in early metestrus, decreased further in length in late metestrus and became very short and rare in anestrus. The variable appearance in early metestrus was not influenced by changing the osmolarity of the fixative and might be a physiological process. The number of glandular openings showed little variability throughout the estrous cycle. Ciliated cells were rare but present in all cycle stages except in late metestrus. However, in the latter cycle stage and in anestrus rare single strands were noted. Transmission electron microscopy was used to determine the inner structure of these strands. Microtubuli were detected in transversal and longitudinal sections but without the 9 + 2 arrangement which is characteristic for cilia. The nature and function of these structures remain unclear.


Assuntos
Endométrio/fisiologia , Endométrio/ultraestrutura , Ciclo Estral/fisiologia , Animais , Cães , Feminino , Microscopia Eletrônica de Varredura
7.
J Endocrinol ; 175(3): 779-92, 2002 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-12475388

RESUMO

Peroxisomes are ubiquitous organelles required for several metabolic functions. Their dysfunction is responsible for a group of human inherited disorders. In the search for endogenous factors regulating the peroxisomal compartment in normal liver, we treated female rats with dehydroepiandrosterone (DHEA) and 25-hydroxycholecalciferol for 1 and 6 days. Relative transcription levels of 39 selected genes were evaluated by real-time quantitative RT-PCR analysis. Catalase (peroxisomal marker)-specific activity was assayed in total liver homogenate and peroxisomes were visualized by catalase localization. DHEA induced peroxisome proliferation and raised catalase specific activity. Expression levels of 16 (of which 11 were peroxisomal) genes were altered. Pex 11, acyl-CoA oxidase,l - andd -multifunctional enzyme, thiolase 1, phytanoyl-CoA hydroxylase, 70 kDa peroxisomal membrane protein and very long chain acyl-CoA synthetase were upregulated, three others were downregulated. Vitamin D caused downregulation of six genes. Administration of vitamin D to peroxisomal disorder patients may be contraindicated. The adrenocortical hormone DHEA is a potential natural regulator of the peroxisomal compartment. Its therapeutic use in X-linked adrenoleukodystrophy, some other beta-oxidation defects and classical Refsum should be considered.


Assuntos
Calcifediol/farmacologia , Desidroepiandrosterona/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Fígado/metabolismo , Peroxissomos/genética , Animais , Sequência de Bases , Catalase/análise , Contraindicações , Primers do DNA/genética , Ativação Enzimática , Feminino , Expressão Gênica , Dados de Sequência Molecular , Transtornos Peroxissômicos/tratamento farmacológico , Peroxissomos/enzimologia , Propilenoglicol/farmacologia , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa
8.
J Histochem Cytochem ; 49(10): 1277-84, 2001 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-11561012

RESUMO

Dissipation of mitochondrial membrane potential (DeltaPsi(m)) and release of cytochrome c from mitochondria appear to be key events during apoptosis. The precise relationship (cause or consequence) between both is currently unclear. We previously showed in a model of serum-free cultured granulosa explants that cytochrome c is retained in a subset of respiring mitochondria until late in the apoptotic process. In this study we further investigated the issue of heterogeneity by using the DeltaPsi(m)-sensitive probe CM-H2TMRos in combination with a DNA fluorochrome. Changes of DeltaPsi(m) were assessed qualitatively by epifluorescence microscopy and were quantified using digital imaging microscopy. This approach yielded the following results: (a) CM-H2TMRos staining is a reliable and specific procedure to detect DeltaPsi(m) changes in granulosa cells explants; (b) dissipation of transmembrane potential is an early event during apoptosis preceding nuclear changes but is confined to a subpopulation of mitochondria within an individual cell; (c) in frankly apoptotic cells a few polarized mitochondria can be detected. These findings support the hypothesis that ATP needed for completion of the apoptotic cascade can be generated during apoptosis in a subset of respiring mitochondria and is not necessarily derived from anaerobic glycolysis.


Assuntos
Apoptose , Mitocôndrias/fisiologia , Animais , Separação Celular , Células Cultivadas , Grupo dos Citocromos c/metabolismo , Feminino , Corantes Fluorescentes , Células da Granulosa/ultraestrutura , Potenciais da Membrana , Microscopia de Fluorescência , Mitocôndrias/enzimologia , Codorniz , Frações Subcelulares/ultraestrutura , Xantenos
9.
Altern Lab Anim ; 29(1): 35-53, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11178573

RESUMO

L-Proline supplementation of the medium for collagen gel cultures of hepatocytes has been shown to improve albumin secretion. A study was made as to whether L-proline is also essential for the maintenance of xenobiotic biotransformation capacities in collagen gel sandwich and immobilisation cultures of rat and human hepatocytes. Key phase I (cytochrome P450-dependent monooxygenase [CYP)] and microsomal epoxide hydrase [mEH]) and phase II (glutathione S-transferase [GST]) biotransformation enzyme activities and the secretion of albumin in the culture medium were assessed in the absence and presence of L-proline. CYP and mEH activities were not affected by the addition of L-proline, whereas phase II alpha-Class GST activity of rat hepatocytes in collagen cultures was decreased. Species differences were demonstrated, as human hepatocytes showed a better maintenance of GST activities than their rat counterparts in the presence of L-proline. Albumin secretion, often considered to be a marker for differentiated cell function, does not parallel the biotransformation capacities of the hepatocytes in culture. Additional results demonstrated an L-proline-mediated enhancement of the proliferation rate of contaminating stellate cells in conventional monolayer culture. Transdifferentiation of stellate cells to proliferating myofibroblasts, along with an increased albumin secretion and collagen synthesis, are characteristic of fibrotic liver. Since the last two phenomena have been observed in L-proline-supplemented collagen gel cultures, it can be concluded that when stable collagen gel cultures of rat hepatocytes are needed for long-term pharmacotoxicological studies, it is preferable to use an L-proline-free culture medium. Further studies on medium optimisation are required for hepatocytes from species other than rat.


Assuntos
Colágeno , Meios de Cultura , Hepatócitos/metabolismo , Prolina/farmacologia , Xenobióticos/metabolismo , Adulto , Idoso , Albuminas/metabolismo , Animais , Biotransformação , Técnicas de Cultura de Células , Sistema Enzimático do Citocromo P-450/metabolismo , Citosol/enzimologia , Epóxido Hidrolases/metabolismo , Feminino , Glutationa Transferase/metabolismo , Hepatócitos/efeitos dos fármacos , Hepatócitos/ultraestrutura , Humanos , Hidroxilação , Masculino , Microssomos Hepáticos/enzimologia , Pessoa de Meia-Idade , Ratos , Ratos Sprague-Dawley , Testosterona/metabolismo
10.
Virchows Arch ; 436(5): 459-65, 2000 May.
Artigo em Inglês | MEDLINE | ID: mdl-10881739

RESUMO

Hyperpipecolic acidaemia is still regarded as a peroxisomal assembly deficiency. The enzyme responsible for the accumulation of pipecolic acid is located in the peroxisomes in man. We studied the appearance and alterations of peroxisomes in liver biopsy material from three unrelated children suffering from isolated hyperpipecolic acidaemia, in which only the metabolism of pipecolic acid is disturbed, using light and electron microscopy after cytochemical staining for visualisation of peroxisomes. Morphometric results showed the presence of normal-sized to small peroxisomes, an increase in number and abnormally shaped organelles, suggesting enhancement of metabolic efficiency. In one case enlarged organelles were observed. Skin fibroblasts were studied in all patients: their peroxisomes appeared to be normal. The obvious presence of peroxisomes in isolated HPA indicates that this disorder should be classified as a single peroxisomal enzyme deficiency.


Assuntos
Hepatopatias/classificação , Transtornos Peroxissômicos/classificação , Peroxissomos/enzimologia , Acil-CoA Oxidase , Alanina Transaminase/metabolismo , Catalase/metabolismo , Células Cultivadas , Criança , Feminino , Fibroblastos/enzimologia , Humanos , Lactente , Hepatopatias/enzimologia , Hepatopatias/patologia , Masculino , Oxirredutases/metabolismo , Transtornos Peroxissômicos/enzimologia , Transtornos Peroxissômicos/patologia , Peroxissomos/ultraestrutura , Pele/citologia , Pele/enzimologia , Transaminases/metabolismo
11.
Cell Death Differ ; 7(4): 331-7, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10773817

RESUMO

Release of apoptogenic factors into the cytosol including cytochrome c is triggering the execution phase of apoptosis through activation of cytoplasmic effector caspases. How loss of function of the electron transport chain can be reconciled with an adequate energy supply necessary for executing the apoptotic program was studied in granulosa cell (GC) sheets cultured up to 72 h without gonadotrophic support. Cytochrome c was localized ultrastructurally by oxidation of diaminobenzidine tetrahydrochloride both in living and fixed cells. In uncultured GC sheets all cells show staining over their entire mitochondrial population. In 72 h cultured sheets in the absence of FSH pre-apoptotic GC's display two subsets of mitochondria: normal sized stained mitochondria and small orthodox mitochondria without demonstrable cytochrome function. Apoptotic cells contain several mitochondria with preservation of respiratory function besides unstained orthodox mitochondria. The cytochrome c containing mitochondria typically display dilated intracristal spaces, a mitochondrial conformation related to increased ATP production. Cytochrome c release was confirmed by Western blotting. In 72 h cultures supplemented with FSH, GC's displayed staining over their entire mitochondrial population. In cultures lacking FSH, but partially protected from apoptosis through caspase inhibition, the cytochrome c release was not inhibited. Thus in the present studied model dysfunction of only a subset of mitochondria is instrumental to initiate the apoptotic program while a functional electron transport chain is maintained until the degradation phase in a subset of respiring mitochondria.


Assuntos
Apoptose , Grupo dos Citocromos c/metabolismo , Células da Granulosa/citologia , Células da Granulosa/fisiologia , Mitocôndrias/metabolismo , Animais , Células Cultivadas , Coturnix , Grupo dos Citocromos c/análise , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Feminino , Células da Granulosa/ultraestrutura , Mitocôndrias/ultraestrutura
12.
J Hepatol ; 32(3): 381-91, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10735606

RESUMO

BACKGROUND/AIMS: Peroxisomes in wild-type cells vary between tissues and developmental stages. In the liver of some peroxisomal deficiency disorder patients, rare parenchymal cells express normal peroxisomes (mosaics); the mechanism is unknown. Our aim was to find factors regulating peroxisome expression. METHODS: Liver-specific as well as peroxisome characteristics were studied in three types of primary rat hepatocyte cultures. RESULTS: Total glutathione S-transferase activity and albumin secretion both increased in the collagen I sandwich and immobilization gel cultures. In contrast, in monolayers cultured on plastic, total glutathione S-transferase activity decreased and albumin secretion was only 30-40% compared to the collagen cultures. Glycogen rosettes typical of liver parenchymal cells were always abundant. Laminin and collagen IV-producing stellate cells were numerous in the monolayer but almost absent in the sandwich cultures. In 6-day-monolayer cultures, the number of liver-specific peroxisomes had decreased while atypical small or elongated peroxisomes appeared. Immunolabeling density for catalase and three beta-oxidation enzymes was decreased compared to adult rat liver; catalase specific activity in homogenates had dropped to 15% and 4% in the sandwich and monolayer cultures, respectively. In 17-day-sandwich cultures, some peroxisomes showed a very weak catalase reaction; total activity was 5%. Supplementation of the collagen type I cultures with several extracellular matrix factors could not prevent peroxisome dedifferentiation. CONCLUSION: The presence of these extracellular matrix components is not sufficient for normal peroxisome expression. It is suggested that hepatocyte-specific and peroxisomal features are regulated differently. The sandwich preserves hepatocyte differentiation better than the monolayer.


Assuntos
Matriz Extracelular/fisiologia , Fígado/metabolismo , Peroxissomos/metabolismo , Animais , Catalase/metabolismo , Células Cultivadas , Colágeno , Técnicas Citológicas , Géis , Glutationa Transferase/metabolismo , Imuno-Histoquímica , Fígado/citologia , Masculino , Peroxissomos/enzimologia , Peroxissomos/ultraestrutura , Ratos , Ratos Sprague-Dawley , Albumina Sérica/metabolismo , Distribuição Tecidual
13.
Ann Neurol ; 47(1): 109-13, 2000 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-10632109

RESUMO

We describe an 18-year-old patient with psychomotor retardation and abnormally short metatarsi and metacarpals but no other signs of classic Refsum disease. Molecular analysis of the phytanoyl-coenzyme A hydroxylase gene revealed a homozygous deletion causing a frameshift. Surprisingly, L-pipecolic acid was elevated in plasma, and microscopy of the liver showed a reduced number of peroxisomes per cell and a larger average peroxisome size. These abnormal peroxisomes lacked catalase as did peroxisomes in fibroblasts of this patient. Such generalized peroxisomal abnormalities are not present in classic Refsum disease.


Assuntos
Erros Inatos do Metabolismo/metabolismo , Oxigenases de Função Mista/metabolismo , Ácido Fitânico/metabolismo , Ácidos Pipecólicos/metabolismo , Doença de Refsum/metabolismo , Criança , Feminino , Humanos , Doença de Refsum/enzimologia , Doença de Refsum/genética
14.
J Histochem Cytochem ; 48(2): 167-78, 2000 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-10639483

RESUMO

Studying the regulation of peroxisome (Px) expression could improve our understanding of human peroxisomal disorders. The granulosa of the largest preovulatory quail follicles proved to be a relevant model because (a) Px expression changes according to the follicular maturation stage and (b) Px expression varies regionally according to the distance of the granulosa relative to the germinal disc region containing the female gamete (oocyte). The question was asked whether Px expression is related to the extent of metabolic cell coupling and whether zonal Px variation is causally related to oocytal factors. This was evaluated by the presence of catalase and Cx-43 (marker proteins for peroxisomes and gap junctions, respectively) and by in vitro experiments with granulosa explants. The data obtained show that the expression of Cx-43 and Px is inversely correlated both temporally and spatially. Uncoupling of gap junctions results in an upregulation of alpha-catalase immunofluorescence. This is in agreement with reports that gap junctions are often negatively affected by Px proliferators. The zonal gradient in Px expression appears to be imposed by the oocyte, as is the case for steroidogenesis and proliferative capacity in the granulosa epithelium. (J Histochem Cytochem 48:167-177, 2000)


Assuntos
Conexina 43/biossíntese , Junções Comunicantes/metabolismo , Células da Granulosa/metabolismo , Células da Granulosa/ultraestrutura , Peroxissomos/metabolismo , Animais , Catalase/metabolismo , Células Cultivadas , Técnicas de Cocultura , Coturnix , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Junções Comunicantes/efeitos dos fármacos , Ácido Glicirretínico/farmacologia , Células da Granulosa/efeitos dos fármacos , Células da Granulosa/enzimologia , Microscopia Eletrônica , Octanóis/farmacologia , Peroxissomos/enzimologia , Fatores de Tempo
15.
J Child Neurol ; 14(7): 434-9, 1999 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10573465

RESUMO

Peroxisomal disorder phenotypes are the result of mutations that cause defective peroxisomal assembly or alterations in the import mechanism of peroxisomal proteins that lead to multiple peroxisomal dysfunctions, or the result of a peroxisomal enzymatic deficiency with a single peroxisomal dysfunction. With complementation analysis, 16 groups have been found. Assignment of the genetic defect has been described for some of the complementation groups. We describe the clinical evolution and follow-up over 10 years of a patient who belongs to complementation group 4, although he showed a milder clinical course. It has been found in fibroblasts different peroxisome populations, normal processing and expression of beta-oxidation PTS1 and PTS2 proteins, abnormal ALD protein distribution and normal plasmalogen biosynthesis; abnormal beta-oxidation metabolites have also been detected in serum. Ultrastructural studies in liver showed peroxisomal mosaicism as in fibroblasts. It has been taken into account that peroxisomal mosaicism may lead to variability in peroxisomal diagnostic parameters, making difficult the final diagnosis in these patients.


Assuntos
Expressão Gênica , Mosaicismo , Transtornos Peroxissômicos/diagnóstico , Transtornos Peroxissômicos/genética , Peroxissomos/genética , Adolescente , Diagnóstico Diferencial , Potenciais Evocados , Teste de Complementação Genética , Humanos , Masculino , Transtornos Peroxissômicos/metabolismo , Transtornos Peroxissômicos/patologia , Peroxissomos/metabolismo , Peroxissomos/patologia , Fenótipo
16.
Brain Res Bull ; 48(6): 555-67, 1999 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10386835

RESUMO

In this article part of the forebrain of the bank vole (Clethrionomys glareolus) is presented in stereotaxic coordinates. The stereotaxic procedure was performed as follows. With the vole's head mounted in a stereotaxic adaptor, internal reference tracks were made with a 0.5-mm diameter microdialysis cannula and India ink, 2 mm in front and 2.6 mm behind the skull landmark bregma. Brains were fixed for 72 h in 4% commercial formaldehyde in sodiumcacodylate buffer containing 1% CaCl2. To determine shrinkage they were weighed before and after fixation. After embedding in paraffin they were sectioned at 25 microm and stained with Nissl. Photomicrographs were taken from the brain of one animal while its frontal (antero-posterior) coordinates of five neural structures were compared with those of 12 other voles. Variability was also checked in lateral and vertical directions at frontal level -1.0 mm (relative to bregma). The results show that the distance between the two skull landmarks bregma and lambda correlates significantly and negatively with the antero-posterior position of each of the brain areas. On the basis of these results an equation is proposed to improve accuracy in locating neural structures that deviate due to biological variability.


Assuntos
Arvicolinae/anatomia & histologia , Prosencéfalo/anatomia & histologia , Técnicas Estereotáxicas , Animais
17.
Toxicol In Vitro ; 13(4-5): 579-85, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-20654519

RESUMO

Collagen gel cultures of hepatocytes represent a promising in vitro model in pharmaco-toxicology. Epidermal growth factor (EGF) is usually added to the culture medium, although one could question its value in a culture model aiming at maintaining a maximum of differentiated functional capacities. In this study, the effects of EGF (20 ng/ml) on albumin secretion, morphology and pentoxyresorufin O-depentylase (PROD), ethoxyresorufin O-deethylase (EROD) and glutathione S-transferase (GST) activities have been examined in both collagen gel sandwich and immobilization gel cultures of adult rat hepatocytes. Transmission electron microscopy did not show an obvious influence of epidermal growth factor (EGF) on the intracellular organization of organelles of the rat hepatocytes. It was found that EGF addition had no effect on albumin secretion in both culture models. On the contrary, the presence of EGF in the culture medium provoked in collagen gel sandwich cultures, after 7 days, significant decreases of 66% and 25% in EROD and PROD activities, respectively. On GST activities, no effect of EGF could be observed in both collagen gel cultures. Removal of EGF from the culture medium seemed to have a positive effect on the maintenance of the phase 1 biotransformation capacity of rat hepatocytes. Its addition should therefore be avoided in collagen gel cultures used in pharmaco-toxicology.

18.
Toxicol In Vitro ; 13(4-5): 571-7, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-20654518

RESUMO

Collagen gel sandwich and immobilization cultures of hepatocytes, using hydrated collagen type I as extracellular matrix (ECM), have been proposed as long-term in vitro models in pharmaco-toxicology. The in vivo ECM composition in the space of Disse is, however, much more complex. As a differentiated hepatocyte phenotype is thought to be highly dependent on ECM composition and biophysical characteristics, we modulated the ECM to mimic the in vivo situation. Moreover, commercially available collagen type I (Boehringer-Ingelheim) was compared to the one prepared in the laboratory from rat tails. ECM composition had no effect on albumin secretion or hepatocyte morphology in both collagen gel sandwich and immobilization cultures. Total, Alpha and Mu class GST activities in organotypical cultures with a complex or a simple collagen type I ECM were similar. The Pi class GST activity increased as a function of culture time in all culture models. Thus, mimicking the in vivo composition of the ECM did not improve the changes in GST expression that were observed in simple collagen gel cultures. The collagen type I matrix is therefore assumed to confer sufficient protection to help the hepatocytes to maintain their differentiated phenotype to a certain extent. Moreover, we hypothesize that the collagen gel matrix may act as a scaffold to keep newly synthesized ECM components in the proximity of the basolateral surfaces of the hepatocytes.

19.
Pediatr Radiol ; 28(10): 790-3, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9799302

RESUMO

We report a male neonate with craniofacial dysmorphic features, multiple congenital anomalies and an unusual form of chondrodysplasia punctata. Radiographic examination revealed punctate epiphyses and coronal clefting of the thoracic spine. The hand radiographs showed some similarities to the brachytelephalangic type of chondrodysplasia punctata. However, the disorder did not fit well with any known entity of chondrodysplasia punctata or other condition characterized by punctate epiphyses.


Assuntos
Anormalidades Múltiplas/diagnóstico por imagem , Condrodisplasia Punctata/diagnóstico por imagem , Anormalidades Múltiplas/genética , Condrodisplasia Punctata/genética , Epífises/diagnóstico por imagem , Face/anormalidades , Humanos , Recém-Nascido , Masculino , Radiografia , Síndrome
20.
Neurology ; 51(5): 1427-32, 1998 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-9818873

RESUMO

OBJECTIVE: Characterization of the defect in a patient presenting a peripheral neuropathy with atypical features of distal motor involvement mimicking Werdnig-Hoffmann disease. PATIENT: Clinical signs included generalized hypotonia and floppiness, absence of stretch reflexes, muscle wasting, lack of head control and lingual fasciculations associated with unaffected facial muscles, and normal intellectual development. RESULTS: Normal muscle histology ruled out Werdnig-Hoffmann disease. Elevated plasma concentrations of very long-chain fatty acids and bile acid intermediates combined with normal plasmalogen levels in erythrocytes suggested defective peroxisomal beta-oxidation directly demonstrated by deficient pristanic acid and partially deficient C26:0 was present oxidation in cultured fibroblasts. Severely impaired pipecolic acid oxidation in liver and phytanic acid oxidation in fibroblasts was present. On light and electron microscopy of the liver tissue, rare peroxisomal membrane ghosts and trilamellar inclusions but absence of peroxisomes was noted. Immunoblot analysis revealed absence of peroxisomal beta-oxidation enzymes in liver tissue but normal results in fibroblasts. Remarkably, expression of the peroxisomal defect in fibroblasts was indicated by the finding of mainly cytoplasmatic catalase, as in liver. Preliminary studies excluded classification of this patient within the large PEX1 complementation group. CONCLUSIONS: The results suggest a novel peroxisome biogenesis disorder involving peroxisomal beta-oxidation as well as phytanic and pipecolic acid oxidation rather than an isolated defect of peroxisomal beta-oxidation. The association of a clinical picture mimicking Werdnig-Hoffmann disease with a novel peroxisomal disorder raises the question of whether investigation for peroxisomal function should be considered in every patient with an enigmatic spinal muscular atrophy-like syndrome.


Assuntos
Microcorpos/fisiologia , Atrofias Musculares Espinais da Infância/diagnóstico , Atrofias Musculares Espinais da Infância/fisiopatologia , Síndrome de Zellweger/diagnóstico , Síndrome de Zellweger/fisiopatologia , Ácidos e Sais Biliares/sangue , Células Cultivadas , Diagnóstico Diferencial , Eritrócitos/metabolismo , Evolução Fatal , Ácidos Graxos não Esterificados/sangue , Feminino , Fibroblastos/metabolismo , Humanos , Lactente , Inteligência , Fígado/metabolismo , Microcorpos/metabolismo , Microcorpos/patologia , Músculo Esquelético/patologia , Ácidos Pipecólicos/sangue , Ácidos Pipecólicos/urina , Atrofias Musculares Espinais da Infância/metabolismo , Síndrome de Zellweger/metabolismo
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