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1.
Support Care Cancer ; 32(8): 500, 2024 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-38985388

RESUMO

PURPOSE: Within families affected by parental cancer, open communication impacts the well-being of parents and their children; however, limited research exists on communication patterns in these families. This sub-study addresses this through the Family-SCOUT study, a multicenter, prospective, interventional, and non-randomized investigation with intervention (IG) and control group (CG). The purpose of this sub-study was to identify and compare the differences in communication patterns between the IG and CG as part of the process evaluation. The research question was addressed in both groups: What communication patterns do healthy parents perceive within their families? METHODS: Using a qualitative approach, the study involved interviewing healthy parents as surrogates for their families. The interviews were audio-recorded, transcribed, and coded using a template analysis. The resulting data were analyzed at the group level. RESULTS: Twenty-three interviews were conducted in the IG and 27 interviews in the CG. The analysis of themes centered on communication patterns as seen in the family structure. Both groups exhibited instances of open communication about fears and wishes as well as the use of child-friendly language when discussing cancer. Notable differences were observed: challenges in open communication with children were sorely reported in CG interviews, and "the illness is discussed when necessary" was sorely described in IG interviews. CONCLUSION: This study underscores the need to address and encourage open communication within families with parental cancer.


Assuntos
Comunicação , Neoplasias , Pais , Humanos , Neoplasias/psicologia , Feminino , Masculino , Pais/psicologia , Adulto , Estudos Prospectivos , Criança , Pessoa de Meia-Idade , Pesquisa Qualitativa , Entrevistas como Assunto , Filho de Pais com Deficiência/psicologia
2.
Phys Rev Lett ; 130(7): 071805, 2023 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-36867799

RESUMO

We search for the dark photon dark matter (DPDM) using a cryogenic millimeter-wave receiver. DPDM has a kinetic coupling with electromagnetic fields with a coupling constant of χ and is converted into ordinary photons at the surface of a metal plate. We search for signal of this conversion in the frequency range 18-26.5 GHz, which corresponds to the mass range 74-110 µeV/c^{2}. We observed no significant signal excess, allowing us to set an upper bound of χ<(0.3-2.0)×10^{-10} at 95% confidence level. This is the most stringent constraint to date and tighter than cosmological constraints. Improvements from previous studies are obtained by employing a cryogenic optical path and a fast spectrometer.

3.
ESMO Open ; 9(6): 103493, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38848662

RESUMO

BACKGROUND: Cancer patients with minor children but also their families suffer from significant psychological distress and comorbidity. Protective factors predicting successful coping are well known. Corresponding systematic interventions are rare and limited by access barriers. We developed a comprehensive family-centered intervention for cancer patients with at least one dependent minor. PATIENTS AND METHODS: Family-SCOUT represents a multicentric, prospective, interventional, and controlled study for families with parental cancer and their minor children. In the intervention group (IG), all family members were addressed using a care and case management approach for nine months. Families in the control group (CG) received standard of care. Participating parents were asked to complete the Hospital-Anxiety-Depression-Scale (HADS) questionnaire at enrolment (T0) and after 9 months (T2). The primary outcome was a clinically relevant reduction of distress in at least one parent per family, measured as minimal important difference (MID) of ≥1.6 in the HADS total score. The percentage of families achieving MID is compared between the IG and CG by exact Fisher's test, followed by multivariate confounder analyses. RESULTS: T0-questionnaire of at least one parent was available for 424 of 472 participating families, T2-questionnaire after 9 months was available for 331 families (IG n = 175, CG n = 156). At baseline, both parents showed high levels of distress (HADS total: sick parents IG: 18.7 ± 8.1; CG: 16.0 ± 7.2; healthy partners: IG: 19.1 ± 7.9; CG: 15.2 ± 7.7). The intervention was associated with a significant reduction in parental distress in the IG (MID 70.4% in at least one parent) compared with the CG (MID 55.8%; P = 0.008). Adjustment for group differences from specific confounders retained significance (P = 0.047). Bias from other confounders cannot be excluded. CONCLUSIONS: Parental cancer leads to a high psychosocial burden in affected families. Significant distress reduction can be achieved through an optimized and structured care approach directed at the family level such as family-SCOUT.


Assuntos
Neoplasias , Pais , Humanos , Feminino , Masculino , Neoplasias/psicologia , Neoplasias/terapia , Estudos Prospectivos , Criança , Adulto , Pais/psicologia , Adaptação Psicológica , Inquéritos e Questionários , Estresse Psicológico/etiologia , Adolescente , Pré-Escolar , Pessoa de Meia-Idade
4.
Dis Esophagus ; 25(5): 373-80, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22050449

RESUMO

Nonerosive reflux disease (NERD) is the most common form of gastroesophageal reflux disease. Patients with NERD have a lower response rate to proton pump inhibitors (PPIs) than patients with erosive esophagitis when gauged from relief of heartburn. Sodium alginate decreases the acidity of refluxate and protects the esophageal mucosa. However, whether the addition of sodium alginate to PPI therapy can improve NERD symptoms remains unknown. Accordingly, the aim of this study was to evaluate the efficacy of adding sodium alginate to basal PPI therapy for NERD. Patients who had experienced heartburn on at least 2 days per week during the 1-month period before entering the study and had no endoscopic mucosal breaks (grade M or N according to Hoshihara's modification of the Los Angeles classification) were randomized to one of two treatments for 4 weeks: omeprazole (20 mg once daily) plus sodium alginate (30 mL four times a day) (group A) or omeprazole (20 mg once daily) alone (group B). Eighty-seven patients were enrolled, and 76 patients were randomly assigned to group A (n = 36) or group B (n = 40). Complete resolution of heartburn for at least 7 consecutive days by the end of treatment was significantly more common in group A (56.7%) than in group B (25.7%). One patient from group A had mild drug-related diarrhea that was not clinically serious. In conclusion, omeprazole combined with sodium alginate was better than omeprazole alone in Japanese patients with NERD.


Assuntos
Alginatos/uso terapêutico , Refluxo Gastroesofágico/tratamento farmacológico , Fármacos Gastrointestinais/uso terapêutico , Omeprazol/uso terapêutico , Inibidores da Bomba de Prótons/uso terapêutico , Adulto , Idoso , Transtornos de Deglutição/tratamento farmacológico , Transtornos de Deglutição/etiologia , Quimioterapia Combinada , Feminino , Refluxo Gastroesofágico/complicações , Ácido Glucurônico/uso terapêutico , Azia/tratamento farmacológico , Azia/etiologia , Ácidos Hexurônicos/uso terapêutico , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Resultado do Tratamento
5.
Nat Cell Biol ; 2(8): 500-6, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10934470

RESUMO

The Drosophila fruitless (fru) gene product Fru has been postulated to be a neural sex-determination factor that directs the development of at least two male-specific characteristics, namely courtship behaviour and formation of the muscle of Lawrence (MOL). The fru gene encodes a putative transcription factor with a BTB domain and two zinc-finger motifs, and with consensus Tra-binding sequences. The binding of Tra to these sequences results in sex-specific alternative splicing of the fru mRNA, leading to production of the 'male-type' or 'female-type' Fru protein. We show here that the Fru protein is not detected in the female central nervous system (CNS), despite the similar level of expression of fru mRNA in both male and female CNS. As ectopic expression of both the 'male-type' (with the sequence for the amino-terminal extension) and 'female-type' (without the sequence for the amino-terminal extension) fru cDNA can induce formation of the MOL in females, the presence or absence of the Fru protein, and not its sex-specific structure, seems to be responsible for the sexually dimorphic actions of the fru gene.


Assuntos
Proteínas de Drosophila , Drosophila melanogaster/genética , Expressão Gênica , Músculos/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Caracteres Sexuais , Diferenciação Sexual/genética , Fatores de Transcrição/metabolismo , Alelos , Sequência de Aminoácidos , Animais , Animais Geneticamente Modificados , Sistema Nervoso Central/citologia , Sistema Nervoso Central/crescimento & desenvolvimento , Sistema Nervoso Central/metabolismo , Clonagem Molecular , Drosophila melanogaster/citologia , Drosophila melanogaster/crescimento & desenvolvimento , Drosophila melanogaster/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Genes Reporter/genética , Imuno-Histoquímica , Hibridização In Situ , Proteínas de Insetos/química , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Masculino , Dados de Sequência Molecular , Desenvolvimento Muscular , Músculos/citologia , Proteínas do Tecido Nervoso/biossíntese , Proteínas do Tecido Nervoso/química , Proteínas do Tecido Nervoso/genética , Neurônios/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Regiões Promotoras Genéticas/genética , Ligação Proteica , Biossíntese de Proteínas , Pupa/citologia , Pupa/genética , Pupa/crescimento & desenvolvimento , Pupa/metabolismo , RNA Mensageiro/análise , RNA Mensageiro/genética , Ribonucleoproteínas/genética , Ribonucleoproteínas/metabolismo , Fatores de Transcrição/biossíntese , Fatores de Transcrição/química , Fatores de Transcrição/genética , Transfecção
6.
Methods Find Exp Clin Pharmacol ; 31(7): 433-41, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19907718

RESUMO

Children exposed to environmental stress in the early neonatal period often develop psychiatric or somatic diseases in adulthood. In the present study in mice, we examined how postpartum stress on the mother influences their pups and thus tried to provide new insight into the management of idiopathic short stature. The dams were exposed to daily 3-h immobilization stress (IS) only for 3 weeks from the day after delivery. When compared to the pups of nonstressed dams (control pups), those of the IS dams (IS pups) showed lower body weight and height, which persisted even into adulthood. Their nutritional status was normal. The IS pups also showed low serum concentrations of insulin-like growth factor I (IGF-I) and poor responses to growth hormone-releasing hormone (GHRH) stimulation on day 22 and were behaviorally hyperactive at 8 weeks. Immunohistochemical analysis demonstrated that the number of pituitary GH-positive cells in response to treatment with GHRH was markedly decreased in the IS pups compared to the control pups. The IS dams did not show apparent behavioral abnormalities except downregulation of glucocorticoid receptor (GR) gene expression in the hippocampus. These results suggest that the perturbation of GH secretion in the pituitary glands is involved in the lifelong growth impairment of the IS pups.


Assuntos
Transtornos do Crescimento/etiologia , Hormônio do Crescimento/metabolismo , Período Pós-Parto/fisiologia , Período Pós-Parto/psicologia , Estresse Psicológico/metabolismo , Estresse Psicológico/psicologia , Animais , Comportamento Animal/fisiologia , Condicionamento Psicológico , Depressão/psicologia , Medo/psicologia , Feminino , Expressão Gênica/fisiologia , Transtornos do Crescimento/fisiopatologia , Elevação dos Membros Posteriores/psicologia , Imobilização , Imuno-Histoquímica , Fator de Crescimento Insulin-Like I/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR , Atividade Motora/efeitos dos fármacos , Fenômenos Fisiológicos da Nutrição
7.
Eur J Pain ; 22(10): 1791-1799, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-29956432

RESUMO

BACKGROUND: This study investigated the characteristics of temperature-related evoked neural activities to baseline skin temperatures on target and adjacent sites using contact heat evoked potentials (CHEPs). METHODS: Contact heat evoked potentials were recorded from 12 normal subjects during three stimuli: target temperatures for "warm", "hot" and "pain" were set at 41, 46 and 51 °C, respectively. The baseline temperature was separately set at 30, 35 and 40 °C under all conditions, and a heat pulse was delivered over the right forearm at 41 °C under the warm condition, at 46 °C under the hot condition and at 51 °C under the pain condition. RESULTS: The N2-P2 amplitude was significantly larger at the 40 °C baseline than at the 30 and 35 °C baselines during the pain condition, whereas no significant differences were observed during the hot and warm conditions. In addition, the effects of an interference warm stimulation to adjacent sites were examined; however, no significant effects were observed. CONCLUSIONS: These results suggest that the priming effects of temperature on CHEPs were only observed under the pain condition, indicating the specificity of thermal pain, as well as a difference in the neural mechanisms responsible for thermal noxious and innocuous processing in human brains. SIGNIFICANCE: This study using CHEPs shows the importance of baseline and target skin temperatures to investigate the characteristics of temperature-related neural activities. This measure may contribute to understanding of warm-, hot-, and pain-related neural activities in human brains.


Assuntos
Potenciais Evocados/fisiologia , Temperatura Alta , Neuralgia/etiologia , Temperatura Cutânea , Feminino , Humanos , Adulto Jovem
8.
J Clin Invest ; 92(3): 1153-60, 1993 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-8376577

RESUMO

Both platelet-derived growth factor (PDGF) A- and B-chains are expressed in mammalian neurons, but their precise roles still remain to be clarified. In the present studies, we examined the expression of two PDGF receptor genes in human tumor cell lines derived from neural crest. The expression of alpha and/or beta PDGF receptors was detected in a wide variety of neural crest-derived human tumor cell lines such as neuroblastoma, primitive neuroectodermal tumor, and Ewing's sarcoma by RNA blot analysis, and confirmed by immunoblot analysis. We have also demonstrated that PDGF receptors on the human neuroblastoma cell lines were biologically functional. Accordingly, chemotactic and mitogenic activities were induced by either PDGF-AA or PDGF-BB in serum-free medium. PDGF isoforms as well as nerve growth factor induced morphological changes showing neuronal cell maturation. Moreover, PDGF coordinately increased the levels of the transcript of the midsize neurofilament gene. The neuroblastoma cell lines also expressed the transcripts of PDGF A- and B-chains. These findings suggest that PDGF isoforms are involved not only in the promotion of the neuroblastoma cell growth, but also in neuronal cell migration, growth, and differentiation in human brain development.


Assuntos
Neuroblastoma/metabolismo , Fator de Crescimento Derivado de Plaquetas/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Receptores do Fator de Crescimento Derivado de Plaquetas/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Quimiotaxia , Humanos , Dados de Sequência Molecular , Crista Neural/patologia , Neuroblastoma/patologia , Proteínas de Neurofilamentos/genética , Oligodesoxirribonucleotídeos/química , Proteínas Proto-Oncogênicas c-sis , RNA Mensageiro/genética , RNA Neoplásico/genética , Células Tumorais Cultivadas
9.
Neuroscience ; 149(2): 446-56, 2007 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-17869442

RESUMO

We studied sensori-motor interaction in the primary (SI) and secondary somatosensory cortex (SII) using magnetoencephalography. Since SII in both hemispheres was activated following unilateral stimulation, we analyzed SIIc (contralateral to stimulation) as well as SIIi (ipsilateral to stimulation). Four tasks were performed in human subjects in which a voluntary thumb movement of the left or right hand was combined with electrical stimulation applied to the index finger of the left or right hand: L(M)-L(S) (movement of the left thumb triggered stimulation to the left finger), L(M)-R(S) (movement of the left thumb triggered electrical stimulation to the right finger), R(M)-R(S) (movement of the right thumb triggered electrical stimulation to the right finger), and R(M)-L(S) (movement of the right thumb triggered electrical stimulation to the left finger). Stimulation to the index finger only (S condition) was also recorded. In SI, the amplitude of N20m and P35m was significantly attenuated in the R(M)-R(S) and L(M)-L(S) tasks compared with the S condition, but that for other tasks showed no change, corresponding to a conventional gating phenomenon. In SII, the R(M)-L(S) task significantly enhanced the amplitude of SIIc but reduced that of SIIi compared with the S condition. The L(M)-L(S) and R(M)-R(S) tasks caused a significant enhancement only in SIIi. The L(M)-R(S) task enhanced the amplitude only in SIIc. The laterality index showed that SII modulation with voluntary movement was more dominant in the hemisphere ipsilateral to movement but was not affected by the side of stimulation. These results provided the characteristics of activities in somatosensory cortices, a simple inhibition in SI but complicated changes in SII depending on the side of movement and stimulation, which may indicate the higher cognitive processing in SII.


Assuntos
Córtex Motor/fisiologia , Córtex Somatossensorial/fisiologia , Adulto , Interpretação Estatística de Dados , Vias Eferentes/fisiologia , Estimulação Elétrica , Feminino , Dedos/inervação , Dedos/fisiologia , Lateralidade Funcional/fisiologia , Mãos/inervação , Mãos/fisiologia , Humanos , Magnetoencefalografia , Masculino , Movimento/fisiologia
10.
J Hum Hypertens ; 21(3): 212-9, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17167525

RESUMO

Recent studies have shown that the converse phenomenon of white-coat hypertension called 'reverse white-coat hypertension' or 'masked hypertension' is associated with poor cardiovascular prognosis. We assessed the hypothesis that this phenomenon may specifically influence left ventricular (LV) structure in treated hypertensive patients. A total of 272 outpatients (mean age, 65 years) with chronically treated essential hypertension and without remarkable white-coat effect were enrolled. Patients were classified into two groups according to office and daytime ambulatory systolic blood pressure (SBP); that is subjects without (Group 1: office SBP > or =daytime SBP, n=149) and with reverse white-coat effect (Group 2: office SBP

Assuntos
Hipertensão/diagnóstico , Hipertrofia Ventricular Esquerda/diagnóstico , Idoso , Monitorização Ambulatorial da Pressão Arterial , Comorbidade , Ecocardiografia , Feminino , Humanos , Hipertensão/tratamento farmacológico , Hipertensão/epidemiologia , Hipertrofia Ventricular Esquerda/epidemiologia , Hipertrofia Ventricular Esquerda/fisiopatologia , Japão/epidemiologia , Masculino , Análise de Regressão , Fatores de Risco
11.
Kyobu Geka ; 59(5): 377-82, 2006 May.
Artigo em Japonês | MEDLINE | ID: mdl-16715888

RESUMO

A small lesion showing ground-glass opacity (GGO) by preoperative computed tomography (CT) is sometimes difficult to detect after lobectomy when it locates in the central part of the lobe. In order to facilitate to identify the lesion for marking pathological specimen, we developed a new method using CT. After surgery, the resected pulmonary lobe was expanded with airflow through the bronchial stump and the target lesion was examined with CT. The laser beam of the CT on the surface of the lung is used as a guiding line for cutting. Through the application of this method for 2 clinical cases, it was found to be possible to exactly identify the GGO lesion from the surface of the resected lung enabling to visualize a fresh surface of the lesion like a CT image with minimal destruction of the structure.


Assuntos
Neoplasias Pulmonares/patologia , Pulmão/diagnóstico por imagem , Pulmão/patologia , Tomografia Computadorizada por Raios X , Idoso , Humanos , Neoplasias Pulmonares/diagnóstico por imagem , Neoplasias Pulmonares/cirurgia , Masculino , Pessoa de Meia-Idade , Pneumonectomia
12.
Neuroscience ; 315: 104-13, 2016 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-26701295

RESUMO

To elucidate the neural substrate associated with capabilities for kinesthetic motor imagery of difficult whole-body movements, we measured brain activity during a trial involving both kinesthetic motor imagery and action observation as well as during a trial with action observation alone. Brain activity was assessed with functional magnetic resonance imaging (fMRI). Nineteen participants imagined three types of whole-body movements with the horizontal bar: the giant swing, kip, and chin-up during action observation. No participant had previously tried to perform the giant swing. The vividness of kinesthetic motor imagery as assessed by questionnaire was highest for the chin-up, less for the kip and lowest for the giant swing. Activity in the primary visual cortex (V1) during kinesthetic motor imagery with action observation minus that during action observation alone was significantly greater in the giant swing condition than in the chin-up condition within participants. Across participants, V1 activity of kinesthetic motor imagery of the kip during action observation minus that during action observation alone was negatively correlated with vividness of the kip imagery. These results suggest that activity in V1 is dependent upon the capability of kinesthetic motor imagery for difficult whole-body movements. Since V1 activity is likely related to the creation of a visual image, we speculate that visual motor imagery is recruited unintentionally for the less vivid kinesthetic motor imagery of difficult whole-body movements.


Assuntos
Imaginação/fisiologia , Desempenho Psicomotor/fisiologia , Córtex Visual/fisiologia , Adulto , Mapeamento Encefálico , Humanos , Cinestesia , Imageamento por Ressonância Magnética , Masculino , Percepção de Movimento/fisiologia , Adulto Jovem
13.
Biochim Biophys Acta ; 1177(1): 93-8, 1993 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-8485173

RESUMO

An antiserum was developed in a rabbit against rat-brain A1 adenosine receptor. This antiserum recognized the denatured form of the purified rat-brain A1 adenosine receptor in immunoblot analysis and the native form of the receptor in the immunoprecipitation analysis. Immunoblot analysis of unpurified or purified adenosine receptor preparations from rat-brain membranes revealed a major immunoreactive band at a position of molecular mass of approx. 35 kDa, which corresponds to the position of purified rat-brain A1 adenosine receptor. Although A1 adenosine receptors from other rat tissues such as testis and adipocyte were also found to be immunoreactive with this antiserum by immunoblot analysis, purified human-brain A1 adenosine receptors showed a poor reactivity with this antibody. The order of the relative immunoreactivity of these A1 adenosine receptors with the antiserum was found to be brain > adipocyte > or = testis. Moreover, the immunoreactivity of these receptors significantly increased after these receptor preparations were deglycosylated by endoglycosidase F. After the deglycosylation, no significant differences in both the immunoreactivity and molecular mass among these receptor preparations were found on the immunoblot. These results suggest that the differences in the molecular mass or immunoreactivity among the A1 adenosine receptor preparations from three rat tissues were mainly due to the difference of sugar moiety present in each receptor molecule. These data are the first to provide analyses of immunological characteristics of A1 adenosine receptors from different tissues and species.


Assuntos
Anticorpos/imunologia , Receptores Purinérgicos/imunologia , Tecido Adiposo/metabolismo , Animais , Sítios de Ligação de Anticorpos , Encéfalo/metabolismo , Masculino , Manosil-Glicoproteína Endo-beta-N-Acetilglucosaminidase , Membranas/metabolismo , Peso Molecular , Testes de Precipitina , Ratos , Receptores Purinérgicos/análise , Receptores Purinérgicos/isolamento & purificação , Testículo/metabolismo
14.
Biochim Biophys Acta ; 614(2): 313-27, 1980 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-7407194

RESUMO

Phenylalanine 4-monooxygenase (L-phenylalanine, tetrahydropteridine:oxygen oxidoreductase (4-hydroxylating), EC 1.14.16.1) was purified approx. 600-fold to apparent homogeneity with a 48% yield from rat liver. Two distinct active forms were separable by calcium phosphate gel chromatography and numbered based on their order of elution from the gel column. The predominant form, Form I, had an estimated molecular weight of about 240 000. The enzyme gave a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis, the molecular weight of which was estimated to be approx. 51 000, indicating that the enzyme might be composed of four identical subunits. The molecular properties of Form I were: sedimentation coefficient, 10.1 S; Stokes radius, 55 A degrees; diffusion coefficient, 3.90 x 10(-7) cm2/s; frictional ratio, 1.33 and isoelectric point, pH 5.6. The enzyme contained approx. 0.6 mol of iron and 0.3 mol of phosphate/mol of subunit of the enzyme. No significant differences in kinetic properties of the two forms, Form I and Form II, were observed. Amino acid analysis studies revealed that the amino acid composition of Form I was essentially identical with that of Form II, indicating that both forms might be the products of the same gene. There were, however, minor differences in the phosphate content and the isoelectric point between the two forms.


Assuntos
Fígado/enzimologia , Fenilalanina Hidroxilase/análise , Aminoácidos/análise , Animais , Cromatografia em Gel , Eletroforese em Gel de Poliacrilamida , Concentração de Íons de Hidrogênio , Cinética , Masculino , Peso Molecular , Fenilalanina Hidroxilase/metabolismo , Ratos
15.
Biochim Biophys Acta ; 527(1): 171-81, 1978 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-102359

RESUMO

The nature of the oxygenated intermediate observed (Fujisawa, H., Hiromi, K., Uyeda, M., Okuno, S., Nozaki, M. and Hayaishi, O. (1972) J. Biol. Chem. 247, 4422--4428) during the reaction of protocatechuate 3,4-dioxygenase (protocatechuate:oxygen 3,4-oxidoreductase (decyclizing), EC 1.13.11.3) was investigated. 3,4-Dihydroxyphenylpropionic acid and 3,4-dihydroxyphenylacetic acid were used as substrates of the enzyme to slow down the rate of the reaction. The enzyme reactions were performed under conditions where the concentration of the organic substrate was lower than those of the enzyme and oxygen in the reaction mixture. The reactions were stopped before completion by the addition of hydrochloric acid or guanidine hydrochloride and then the organic compounds were extracted from the reaction mixture to be analyzed. The qualitative analyses by thin-layer chromatography revealed that there was no species other than the organic substrate and the enzymatic reaction end-product during reaction. The quantitative spectrophotometric analyses revealed that the organic substrate which had participated in the formation of the oxygenated intermediate existed as a species indistinguishable from the reaction end-product, indicating that the oxygenated intermediate was not a simple complex of oxygen, substrate and the enzyme, i.e., a ternary complex, but a species rather close to a binary complex of product and the enzyme.


Assuntos
Oxigenases/metabolismo , Protocatecoate-3,4-Dioxigenase/metabolismo , Ácido 3,4-Di-Hidroxifenilacético , Ácidos Cafeicos , Catecóis , Cinética , Consumo de Oxigênio , Fenilpropionatos , Ligação Proteica , Pseudomonas aeruginosa/enzimologia , Espectrofotometria , Espectrofotometria Ultravioleta
16.
J Gen Physiol ; 116(2): 147-61, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10919863

RESUMO

Cyclic AMP is a ubiquitous second messenger that coordinates diverse cellular functions. Current methods for measuring cAMP lack both temporal and spatial resolution, leading to the pervasive notion that, unlike Ca(2+), cAMP signals are simple and contain little information. Here we show the development of adenovirus-expressed cyclic nucleotide-gated channels as sensors for cAMP. Homomultimeric channels composed of the olfactory alpha subunit responded rapidly to jumps in cAMP concentration, and their cAMP sensitivity was measured to calibrate the sensor for intracellular measurements. We used these channels to detect cAMP, produced by either heterologously expressed or endogenous adenylyl cyclase, in both single cells and cell populations. After forskolin stimulation, the endogenous adenylyl cyclase in C6-2B glioma cells produced high concentrations of cAMP near the channels, yet the global cAMP concentration remained low. We found that rapid exchange of the bulk cytoplasm in whole-cell patch clamp experiments did not prevent the buildup of significant levels of cAMP near the channels in human embryonic kidney 293 (HEK-293) cells expressing an exogenous adenylyl cyclase. These results can be explained quantitatively by a cell compartment model in which cyclic nucleotide-gated channels colocalize with adenylyl cyclase in microdomains, and diffusion of cAMP between these domains and the bulk cytosol is significantly hindered. In agreement with the model, we measured a slow rate of cAMP diffusion from the whole-cell patch pipette to the channels (90% exchange in 194 s, compared with 22-56 s for substances that monitor exchange with the cytosol). Without a microdomain and restricted diffusional access to the cytosol, we are unable to account for all of the results. It is worth noting that in models of unrestricted diffusion, even in extreme proximity to adenylyl cyclase, cAMP does not reach high enough concentrations to substantially activate PKA or cyclic nucleotide-gated channels, unless the entire cell fills with cAMP. Thus, the microdomains should facilitate rapid and efficient activation of both PKA and cyclic nucleotide-gated channels, and allow for local feedback control of adenylyl cyclase. Localized cAMP signals should also facilitate the differential regulation of cellular targets.


Assuntos
Adenilil Ciclases/análise , Adenilil Ciclases/metabolismo , AMP Cíclico/farmacocinética , Canais Iônicos/análise , Canais Iônicos/metabolismo , Adenoviridae/genética , Técnicas Biossensoriais/métodos , Cálcio/metabolismo , Sinalização do Cálcio/fisiologia , Compartimento Celular/fisiologia , Células Cultivadas , GMP Cíclico/análogos & derivados , GMP Cíclico/farmacologia , Canais de Cátion Regulados por Nucleotídeos Cíclicos , Citosol/química , Citosol/enzimologia , Diálise , Difusão , Regulação Viral da Expressão Gênica , Humanos , Canais Iônicos/genética , Rim/citologia , Cloreto de Magnésio/farmacologia , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/fisiologia , Modelos Biológicos , Técnicas de Patch-Clamp , Inibidores da Agregação Plaquetária/farmacologia , Tionucleotídeos/farmacologia , Transfecção
17.
APMIS ; 113(7-8): 550-4, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-16086826

RESUMO

Micropapillary carcinoma has been described in various organs, including the breast, urinary bladder, ovary and lung. We here present a case of pulmonary micropapillary carcinoma in a 72-year-old Japanese man who died of respiratory failure and septic shock, following which autopsy was performed. A mass measuring 2.5 x 2.5 x 2.5 cm was observed in the left lower lobe of the lung. The tumor showed moderately differentiated papillary adenocarcinoma with a focal micropapillary component. Carcinomatous lymphangiosis was also observed in the left lung and metastatic lesions were observed in the bilateral lung, liver, vertebra, muscle layer of the urinary bladder, right adrenal gland, spleen and lymph nodes. The micropapillary component was predominant at some metastatic sites. Immunohistochemically, both the adenocarcinoma and micropapillary components were positive for cytokeratin (CK) 7, CK19, TTF (thyroid transcription factor)-1, carcinoembryonic antigen (CEA) and surfactant apoprotein A (SP-A), and negative for CK20, estrogen receptor, progesterone receptor, uroplakin III, and CA125. The invasive area of the conventional adenocarcinoma component contained a large number of myofibroblasts, whereas the stroma of the micropapillary component contained a small number of myofibroblasts. However, no myofibroblasts were observed in the stroma of the central core of the non-invasive micropapillary carcinoma. Several lymphatic invasions by neoplastic cells were identified in the peripheral area of the micropapillary component using D2-40 antibody. The immunohistochemical profile may be helpful in determining the primary location of the neoplasm containing micropapillary features. Myofibroblasts are present in the stroma of the invasive neoplastic nests in the micropapillary component as well as the conventional adenocarcinoma component, and D2-40 monoclonal antibody may be useful for evaluating the lymphatic invasion of pulmonary micropapillary carcinoma.


Assuntos
Adenocarcinoma Papilar/metabolismo , Adenocarcinoma Papilar/patologia , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Idoso , Antígeno Carcinoembrionário/metabolismo , Humanos , Imuno-Histoquímica , Queratina-7 , Queratinas/metabolismo , Masculino , Proteínas Nucleares/metabolismo , Proteína A Associada a Surfactante Pulmonar/metabolismo , Fator Nuclear 1 de Tireoide , Fatores de Transcrição/metabolismo
18.
Cell Signal ; 10(3): 211-6, 1998 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-9607145

RESUMO

GTPase-activating protein (GAP) enhances the intrinsic GTPase activity of cellular Ras. In addition to two Src homology 2 (SH2) domains and one Src homology 3 (SH3) domain, it contains a pleckstrin homology (PH) domain. The wild-type or point mutant in the PH domain of p120 GAP (W568A) was expressed by using the baculovirus/Sf9 cell system. Direct effects of the G protein beta gamma subunit (G beta gamma) and several sphingolipids and the effects of phosphorylation by c-Src on the GTPase-stimulating activity of these GAPs on Ras were examined by using immunoprecipitates of these GAPs. The activities of neither of these GAPs were affected by the addition of G beta gamma, although the W568A mutant bound less to G beta gamma compared with the wild type. Several sphingolipids had no effect on the activity of these GAPs. Only in the W568A mutant was GTPase-stimulating activity reduced by tyrosine phosphorylation by c-Src.


Assuntos
GTP Fosfo-Hidrolases/metabolismo , Fosfoproteínas , Mutação Puntual/fisiologia , Proteínas/metabolismo , Homologia de Sequência de Aminoácidos , Proteínas ras/metabolismo , Proteínas Sanguíneas/genética , GTP Fosfo-Hidrolases/genética , Proteínas de Ligação ao GTP/fisiologia , Proteínas Ativadoras de GTPase , Humanos , Proteína Oncogênica pp60(v-src)/metabolismo , Fosforilação , Proteínas/genética , Proteínas Recombinantes de Fusão , Esfingolipídeos/farmacologia , Tirosina/metabolismo , Proteínas Ativadoras de ras GTPase
19.
Cell Signal ; 9(1): 37-40, 1997 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-9067627

RESUMO

Protein phosphorylation is one of several representative post-translational modifications. Cyclic AMP-dependent protein kinase (PKA) plays the crucial and varying role of signal transduction. On the other hand, ras proteins plays an important role in cell proliferation and growth. Although a previous report showed that H-ras protein was phosphorylated by PKA, the stoichiometry was not determined, so we investigated the stoichiometry of phosphorylation of the protein by PKA. H-ras cDNA inserted into a pGEX-2T expressing vector produced high levels of recombinant H-ras (rH-ras) in a fusion protein with glutathione S-transferase. rH-ras was obtained after cleavage by thrombin. Phosphorylation of ras protein by the catalytic subunit of PKA was performed, and the radioactivity was counted after SDS-PAGE and autoradiography. The results indicate that less than 0.1 mol of phosphate was incorporated per mol of H-ras protein, and suggest that H-ras protein could not be a physiologically meaningful substrate for PKA.


Assuntos
Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Proteínas ras/metabolismo , Fosforilação
20.
Hypertension ; 20(4): 501-10, 1992 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-1398885

RESUMO

Not only blood pressure but also behavioral activity, brain morphology, and cerebral ventricular size differ between young spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats. This suggests that cerebral blood flow and cerebral metabolism may vary between these two rat strains. To test this hypothesis, we measured local cerebral glucose utilization in 31 brain areas of 26-30-week-old rats. Local cerebral blood flow was also assessed in these same areas. Cerebral glucose utilization was measured by the 2-deoxyglucose method; cerebral blood flow was determined by the iodoantipyrene method. In virtually all gray matter structures, the apparent rate of glucose utilization was lower in SHR than in normotensive WKY rats; the interstrain differences varied significantly among structures and were statistically significant (uncorrected t tests) in 14 of 28 gray matter areas. Local cerebral blood flow was fairly similar in the two rat strains. The coupling of blood flow to glucose utilization varied significantly among brain areas in normotensive WKY rats as well as in SHR. In a number of gray matter structures, the coupling of flow to metabolism differed between hypertensive and normotensive animals. These data suggest that for many brain areas, either glucose utilization or glucose partitioning differs between WKY rats and SHR.


Assuntos
Encéfalo/metabolismo , Circulação Cerebrovascular , Glucose/metabolismo , Hipertensão/fisiopatologia , Ratos Endogâmicos SHR/fisiologia , Ratos Endogâmicos WKY/fisiologia , Análise de Variância , Animais , Autorradiografia , Comportamento Animal , Pressão Sanguínea , Temperatura Corporal , Peso Corporal , Dióxido de Carbono/sangue , Hematócrito , Masculino , Concentração Osmolar , Oxigênio/sangue , Ratos
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