RESUMO
To search for low-energy resonant structures in isospin T=3/2 three-body systems, we have performed the experiments ^{3}H(t,^{3}He)3n and ^{3}He(^{3}He,t)3p at intermediate energies. For the 3n experiment, we have newly developed a thick Ti-^{3}H target that has the largest tritium thickness among targets of this type ever made. The 3n experiment for the first time covered the momentum-transfer region as low as 15 MeV/c, which provides ideal conditions for producing fragile systems. However, in the excitation-energy spectra we obtained, we did not observe any distinct peak structures. This is in sharp contrast to tetraneutron spectra. The distributions of the 3n and 3p spectra are found to be similar, except for the displacement in energy due to Coulomb repulsion. Comparisons with theoretical calculations suggest that three-body correlations exist in the 3n and 3p systems, although not enough to produce a resonant peak.
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In an emulsion-counter hybrid experiment performed at J-PARC, a Ξ^{-} absorption event was observed which decayed into twin single-Λ hypernuclei. Kinematic calculations enabled a unique identification of the reaction process as Ξ^{-}+^{14}Nâ_{Λ}^{10}Be+_{Λ}^{5}He. For the binding energy of the Ξ^{-} hyperon in the Ξ^{-}-^{14}N system a value of 1.27±0.21 MeV was deduced. The energy level of Ξ^{-} is likely a nuclear 1p state which indicates a weak ΞN-ΛΛ coupling.
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The cross sections of the ^{7}Be(n,α)^{4}He reaction for p-wave neutrons were experimentally determined at E_{c.m.}=0.20-0.81 MeV slightly above the big bang nucleosynthesis (BBN) energy window for the first time on the basis of the detailed balance principle by measuring the time-reverse reaction. The obtained cross sections are much larger than the cross sections for s-wave neutrons inferred from the recent measurement at the n_TOF facility in CERN, but significantly smaller than the theoretical estimation widely used in the BBN calculations. The present results suggest the ^{7}Be(n,α)^{4}He reaction rate is not large enough to solve the cosmological lithium problem, and this conclusion agrees with the recent result from the direct measurement of the s-wave cross sections using a low-energy neutron beam and the evaluated nuclear data library ENDF/B-VII.1.
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We performed quantitative trait locus (QTL) analyses for egg production traits, including age at first egg (AFE) and egg production rates (EPR) measured every 4 weeks from 22 to 62 weeks of hen age, in a population of 421 F(2) hens derived from an intercross between the Oh-Shamo (Japanese Large Game) and White Leghorn breeds of chickens. Simple interval mapping revealed a main-effect QTL for AFE on chromosome 1 and four main-effect QTL for EPR on chromosomes 1 and 11 (three on chromosome 1 and one on chromosome 11) at the genome-wide 5% levels. Among the three EPR QTL on chromosome 1, two were identified at the early stage of egg laying (26-34 weeks of hen age) and the remaining one was discovered at the late stage (54-58 weeks). The alleles at the two EPR QTL derived from the Oh-Shamo breed unexpectedly increased the trait values, irrespective of the Oh-Shamo being inferior to the White Leghorn in the trait. This suggests that the Oh-Shamo, one of the indigenous Japanese breeds, is an untapped resource that is important for further improvement of current elite commercial laying chickens. In addition, six epistatic QTL were identified on chromosomes 2, 4, 7, 8, 17 and 19, where none of the above main-effect QTL were located. This is the first example of detection of epistatic QTL affecting egg production traits. The main and epistatic QTL identified accounted for 4-8% of the phenotypic variance. The total contribution of all QTL detected for each trait to the phenotypic and genetic variances ranged from 4.1% to 16.9% and from 11.5% to 58.5%, respectively.
Assuntos
Galinhas/genética , Ovos , Locos de Características Quantitativas , Animais , Galinhas/fisiologia , Mapeamento Cromossômico , Cruzamentos Genéticos , Feminino , MasculinoRESUMO
We report two distinct growth modes of pentacene (PEN) and perfluoropentacene (PFP) films on a Bi(0001) substrate investigated by scanning tunneling microscopy (STM). PEN grows epitaxially on Bi(0001) at room temperature (RT), resulting in the formation of bulk-like crystalline films. In contrast, submonolayer PFP forms a two-dimensional (2D) liquid-like phase with PFP molecules loosely bound on Bi(0001). Beyond one monolayer, the PFP molecules diffuse over very long distances to aggregate into three-dimensional (3D) islands, leading to a rough film morphology. Utilizing the stacking interaction at the PFP/PEN interface, we deposited PFP on the template of an ordered PEN monolayer formed on Bi. It is found that PFP molecules nucleate into ordered crystalline islands with PFP molecules standing-up. The different morphologies of PEN and PFP overlayers can be understood in terms of perfluorination induced decoupling of PFP molecules from the Bi substrate below.
Assuntos
Bismuto/química , Cristalização/métodos , Fluorocarbonos/química , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Nanotecnologia/métodos , Naftacenos/química , Substâncias Macromoleculares/química , Teste de Materiais , Conformação Molecular , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
Addition of an electron energy filter to low energy electron microscopy (LEEM) and photoelectron emission microscopy (PEEM) instruments greatly improves their analytical capabilities. However, such filters tend to be quite complex, both electron optically and mechanically. Here we describe a simple energy filter for the existing IBM LEEM/PEEM instrument, which is realized by adding a single scanning aperture slit to the objective transfer optics, without any further modifications to the microscope. This energy filter displays a very high energy resolution ΔE/E = 2 × 10(-5), and a non-isochromaticity of â¼0.5 eV/10 µm. The setup is capable of recording selected area electron energy spectra and angular distributions at 0.15 eV energy resolution, as well as energy filtered images with a 1.5 eV energy pass band at an estimated spatial resolution of â¼10 nm. We demonstrate the use of this energy filter in imaging and spectroscopy of surfaces using a laboratory-based He I (21.2 eV) light source, as well as imaging of Ag nanowires on Si(001) using the 4 eV energy loss Ag plasmon.
RESUMO
The diffusion process of fluorine (F) atoms on the Si(111)-(7x7) surface is investigated using high-temperature scanning tunneling microscopy. The kinetic parameters of F hopping agree well with those of the diffusing silicon (Si) atoms, which implies that of all reaction processes, the Si diffusion serves as the rate-determining one. Deposition of Si on the surface is found to enhance F hopping, which supports the above-mentioned observation. Theory reveals that the replacement of F adsorption sites by diffusing Si atoms is the key process in the diffusion mechanism.
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BACKGROUND: Functional dyspepsia (FD), a heterogeneous disorder, involves multiple pathogenetic mechanisms. Developing treatments for FD has been challenging. We performed a randomized, placebo-controlled, double-blind clinical trial to determine the efficacy of rikkunshito, a Japanese herbal medicine, in FD patients. METHODS: FD patients (n = 192) who met the Rome III criteria without Helicobacter pylori infection, predominant heartburn, and depression were enrolled at 56 hospitals in Japan. After 2 weeks of single-blind placebo treatment, 128 patients with continuous symptoms were randomly assigned to 8 weeks of rikkunshito (n = 64) or placebo (n = 61). The primary efficacy endpoint was global assessment of overall treatment efficacy (OTE). The secondary efficacy endpoints were improvements in upper gastrointestinal symptoms evaluated by the Patient Assessment of Upper Gastrointestinal Disorders-Symptom Severity Index (PAGI-SYM), the Global Overall Symptom scale (GOS), and the modified Frequency Scale for the Symptoms of Gastroesophageal Reflux Disease (m-FSSG), and psychological symptoms evaluated by the Hospital Anxiety and Depression Scale (HADS). KEY RESULTS: Rikkunshito increased OTE compared to placebo at 8 weeks (P = .019). Rikkunshito improved upper gastrointestinal symptoms (PAGI-SYM, GOS, and m-FSSG) at 8 weeks, especially postprandial fullness/early satiety (P = .015 and P = .001) and bloating (P = .007 and P = .002) of the PAGI-SYM subscales at 4 weeks and 8 weeks. Improvement of HADS at 8 weeks (P = .027) correlated with those of PAGI-SYM (r = .302, P = .001), GOS (r = .186, P = .044), and m-FSSG (r = .462, P < .001), postprandial fullness/early satiety (r = .226, P = .014), dyspepsia (r = .215, P = .019), and PDS (r = .221, P = .016). CONCLUSION & INFERENCES: Rikkunshito may be beneficial for FD patients to simultaneously treat gastrointestinal and psychological symptoms.
Assuntos
Ansiedade/diagnóstico , Ansiedade/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Dispepsia/diagnóstico , Dispepsia/tratamento farmacológico , Adulto , Idoso , Idoso de 80 Anos ou mais , Ansiedade/epidemiologia , Método Duplo-Cego , Dispepsia/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Método Simples-Cego , Resultado do Tratamento , Adulto JovemRESUMO
BACKGROUND: Enteric glial cells (EGCs) play important roles in enteric integrity and regulation of gastrointestinal function. However, whether EGCs undergo pathophysiological changes in stress-associated gastrointestinal disorders is unknown. We investigated structural and functional alterations in colonic EGCs and their roles in colonic contraction in an irritable bowel syndrome (IBS) model. METHODS: As a chronic stress, male Wistar rats underwent 3-h maternal separation during postnatal days 2-14. As an acute stress, we used water-immersion stress (4 h) in adulthood (at 8 weeks). We quantitatively and morphologically evaluated enteric neurons and EGCs using whole-mount longitudinal muscle-myenteric plexus preparations. Colonic contraction was analyzed with electrical field stimulation (EFS). KEY RESULTS: Glial fibrillary acidic protein (GFAP) expression and the number of total, cholinergic, and nitrergic neurons were unchanged in maternally separated rats with acute stress (combined stress: an IBS model) compared with controls. However, the density of GFAP-positive EGC processes that apparently overlapped with the neurons and the extent of bulbous swelling of terminals increased according to the stress intensity: control, acute stress, maternal separation, and combined stress. EFS-induced colonic contractions were significantly greater in the combined stress rats than in controls. Higher dose of fluorocitrate, a selective inhibitor of EGC metabolism, was required to inhibit both EFS-induced contraction and EGCs activation in the combined stress rats than in controls. CONCLUSIONS & INFERENCES: Colonic EGCs exhibited structural alterations according to the stress intensity. EGCs were associated with stress-induced colonic hyper-contraction in the combined stress rats, which may underlie the pathogenesis of IBS.
Assuntos
Colo/metabolismo , Motilidade Gastrointestinal/fisiologia , Plexo Mientérico/metabolismo , Neuroglia/metabolismo , Estresse Psicológico/metabolismo , Acetilcolina/metabolismo , Animais , Neurônios Colinérgicos/metabolismo , Colo/fisiopatologia , Proteína Glial Fibrilar Ácida/metabolismo , Masculino , Privação Materna , Plexo Mientérico/fisiopatologia , Óxido Nítrico/metabolismo , Ratos , Ratos Wistar , Estresse Psicológico/fisiopatologiaRESUMO
Fracture repair requires the involvement of osteoclasts (OC), multinucleated cells which are responsible for bone resorption and form by fusion of circulating mononuclear haemopoietic precursors. The nature of these circulating precursor cells, in particular their relationship to blood monocytes, is uncertain. To define further the nature of the circulating human OC precursor, and to determine the role bone stromal cells and humoral factors play in the differentiation of OCs, we co-cultured human umbilical cord blood monocytes with UMR106.01 osteoblast-like cells in the presence and absence of 1,25 dihydroxyvitamin D3 [1,25 (OH)2D3], macrophage-colony stimulating factor (M-CSF) and dexamethasone on both bone slices and coverslips. Isolated cells were positive only for monocyte/macrophage markers (CD11a, CD11b, CD14 and HLA-DR) and negative for OC markers [tartrate resistant acid phosphatase (TRAP), vitronectin receptors (VNR) and calcitonin receptors (CT receptors)] and did not form resorption pits on bone slices after 24 hr in culture. However, after 14 days in co-culture with UMR106.01 cells, in the presence of 1,25 (OH)2D3 and M-CSF, numerous TRAP, CT receptor and VNR positive multinucleated cells capable of extensive lacunar bone resorption were formed in these co-cultures. The presence of 1,25 (OH)2D3, M-CSF and a bone-derived stromal cell population were absolute requirements for OC differentiation. It is concluded that mononuclear phagocytes are capable of differentiating into mature functional OCs when cultured under specific cellular and hormonal conditions. This is vitro model of human OC differentiation should prove useful in further analysing factors controlling OC generation in bone remodelling and repair.
Assuntos
Calcitriol/farmacologia , Sangue Fetal/citologia , Fator Estimulador de Colônias de Macrófagos/farmacologia , Monócitos/citologia , Monócitos/efeitos dos fármacos , Osteoblastos/fisiologia , Osteoclastos/citologia , Animais , Reabsorção Óssea/etiologia , Reabsorção Óssea/patologia , Reabsorção Óssea/fisiopatologia , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Feminino , Histocitoquímica , Humanos , Técnicas In Vitro , Monócitos/metabolismo , Osteoclastos/efeitos dos fármacos , Osteoclastos/metabolismo , Gravidez , Ratos , Receptores da Calcitonina/metabolismoRESUMO
The osteoclast is known to be formed by fusion of circulating mononuclear precursor cells of haematopoietic origin. The precise nature of these circulating cells and, in particular, their relation to monocytes is unknown. We have developed an in vitro system of human osteoclast formation whereby human monocytes [CD14, CD11a, CD11b and HLA-DR positive, and tartrate-resistant acid phosphatase (TRAP), calcitonin receptor (CTR), vitronectin receptor (VNR) negative] were isolated and cocultured for up to 21 days with UMR106 rat osteoblast-like cells or ST2 mouse preadipocytic bone marrow stromal cells in the presence of 1 alpha, 25 dihydroxyvitamin D3 (1,25(OH)2D3) and macrophage colony stimulating factor (M-CSF). Numerous TRAP, VNR and CTR positive multinucleated cells, capable of extensive lacunar bone resorption, formed in these cocultures; the absolute requirements for this to occur were contact with the above bone stromal cells, 1,25(OH)2D3, and M-CSF. These results show that the human mononuclear osteoclast precursor circulates in the monocyte fraction and exhibits a monocyte phenotype, acquiring osteoclast phenotypic features in the process of differentiation into mature functional osteoclasts.
Assuntos
Monócitos/fisiologia , Osteoclastos/fisiologia , Células-Tronco/fisiologia , Fosfatase Ácida/metabolismo , Animais , Medula Óssea/fisiologia , Células da Medula Óssea , Calcitriol/farmacologia , Diferenciação Celular , Técnicas de Cocultura , Humanos , Isoenzimas/metabolismo , Fator Estimulador de Colônias de Macrófagos/farmacologia , Camundongos , Monócitos/citologia , Osteoblastos/fisiologia , Ratos , Receptores da Calcitonina/metabolismo , Receptores de Vitronectina/metabolismo , Células Estromais/citologia , Células Estromais/fisiologia , Fosfatase Ácida Resistente a TartaratoRESUMO
The long-term effectiveness of zonisamide (ZNS) was evaluated in 11 patients with West syndrome (7 symptomatic) who had cessation of spasms with ZNS monotherapy. During the follow-up period (24 to 79 months, mean = 53 months), this response was maintained in 7 patients (3 symptomatic, relapse rate = 36%), including 2 children in whom ZNS was successfully discontinued. No serious adverse reactions were noted. ZNS may be both effective and well tolerated for the treatment of West syndrome.
Assuntos
Anticonvulsivantes/uso terapêutico , Isoxazóis/uso terapêutico , Espasmos Infantis/tratamento farmacológico , Adolescente , Anticonvulsivantes/efeitos adversos , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Lactente , Isoxazóis/efeitos adversos , Assistência de Longa Duração/estatística & dados numéricos , Masculino , Estudos Prospectivos , ZonisamidaRESUMO
Macrophage-colony stimulating factor (M-CSF) is an essential requirement for human osteoclast formation, but its effect on the proliferation and differentiation of circulating osteoclast precursor cells is unknown. Other growth factors and cytokines are also known to support/stimulate osteoclast formation from mouse marrow precursors, but it is not certain whether these factors similarly influence human osteoclast formation. In this study, human monocytes were cocultured with osteoblast-like UMR-106 cells on coverslips and dentine slices for up to 21 days in the presence of 1,25 dihydroxyvitamin D(3) (10(-7) mol/L), dexamethasone (10(-8) mol/L), and various concentrations of either M-CSF or other humoral factors (interleukin [IL]-1beta, IL-3, IL-6, and IL-11; tumor necrosis factor-alpha [TNF-alpha]; and granulocyte macrophage [GM]-CSF). The effect on osteoclast formation was assessed by tartrate-resistant acid phosphatase (TRAP) and vitronectin receptor staining and lacunar bone resorption. The results of time-course and proliferation studies showed that M-CSF stimulated both the proliferative and differentiation stages of human osteoclast formation from circulating osteoclast precursors in a dose-dependent manner. A high concentration of M-CSF (100 ng/mL) did not inhibit osteoclast formation. IL-3 and GM-CSF were also capable of stimulating human osteoclast formation, although these growth factors were much less potent than M-CSF. IL-3- and GM-CSF-stimulated osteoclast formation was inhibited by an antibody specific for human M-CSF. Osteoclast formation and lacunar resorption was not seen when either TNF-alpha, IL-1beta, IL-6 (+ soluble IL-6 receptor), or IL-11 was substituted for M-CSF during coculture. These results confirm that M-CSF is essential for human osteoclast formation from circulating mononuclear precursors, and also shows that IL-3 and GM-CSF may support osteoclast differentiation via the stimulation of M-CSF production by human monocytes.
Assuntos
Divisão Celular/efeitos dos fármacos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Interleucinas/farmacologia , Fator Estimulador de Colônias de Macrófagos/farmacologia , Osteoclastos/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia , Adulto , Humanos , Osteoclastos/citologiaRESUMO
Interleukin-6 (IL-6) and interleukin-11 (IL-11) are known to influence osteoclast formation and bone resorption. In order to determine whether IL-6 and IL-11 could independently support human osteoclast formation, these factors were added to cultures of human peripheral blood mononuclear cells of the monocyte (CD14(+)) fraction in the presence of macrophage colony-stimulating factor (M-CSF). Under these conditions, IL-6 and IL-11 induced the formation of multinucleated cells which were positive for TRAP, VNR, and calcitonin receptor and capable of lacunar resorption. Osteoclastogenesis induced by IL-6 and IL-11 was inhibited by the addition of an anti-gp130 antibody but not by osteoprotegerin. These results indicate that IL-6 and IL-11, which are thought to play a role in several osteolytic bone disorders, are directly capable of inducing osteoclast formation by a RANKL-independent mechanism.
Assuntos
Proteínas de Transporte/fisiologia , Interleucina-11/farmacologia , Interleucina-6/farmacologia , Glicoproteínas de Membrana/fisiologia , Osteoclastos/citologia , Osteoclastos/fisiologia , Fosfatase Ácida/biossíntese , Adulto , Células Cultivadas , Feminino , Humanos , Isoenzimas/biossíntese , Masculino , Pessoa de Meia-Idade , Osteoclastos/ultraestrutura , Ligante RANK , Receptor Ativador de Fator Nuclear kappa-B , Proteínas Recombinantes/farmacologia , Fosfatase Ácida Resistente a TartaratoRESUMO
Transforming growth factor beta (TGFbeta) is a multifunctional growth factor that is produced by many cells in bone and is abundant in the bone matrix. TGFbeta is known to regulate RANKL-induced osteoclast formation and bone resorbing activity. In this study we sought to determine whether TGFbeta could directly induce osteoclast formation by a RANKL-independent mechanism. We found that the addition of TGFbeta to cultures of human monocytes and RAW 264.7 cells (in the presence of M-CSF and the absence of RANKL, TNFalpha or IL-6/IL-11) was sufficient to induce the formation of TRAP+ and VNR+ cells, which formed actin rings and were capable of extensive lacunar resorption. The addition of osteoprotegerin or antibodies to TNFalpha and its receptors, as well as antibodies to gp130, did not inhibit lacunar resorption, indicating that TGFbeta did not act by stimulating RANKL, TNF or IL-6 production by monocytes. TGFbeta-induced osteoclast formation was qualitatively different from that induced by RANKL with numerous TRAP+/VNR+ mononuclear and small multinucleated cells being formed; these cells produced many small resorption lacunae. Our results indicate that TGFbeta, which is abundant in the bone matrix, can, in the presence of M-CSF, directly induce mononuclear phagocyte osteoclast precursors to differentiate into osteoclastic cells capable of lacunar resorption.
Assuntos
Glicoproteínas de Membrana/deficiência , Osteoclastos/citologia , Osteoclastos/efeitos dos fármacos , Fator de Crescimento Transformador beta/farmacologia , Actinas/metabolismo , Antígenos CD/imunologia , Antígenos CD/metabolismo , Proteínas de Transporte/metabolismo , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Receptor gp130 de Citocina , Glicoproteínas/metabolismo , Humanos , Interleucina-1/farmacologia , Receptores de Lipopolissacarídeos/metabolismo , Glicoproteínas de Membrana/antagonistas & inibidores , Glicoproteínas de Membrana/imunologia , Glicoproteínas de Membrana/metabolismo , Microscopia Eletrônica de Varredura , Osteoclastos/metabolismo , Osteoprotegerina , Ligante RANK , Receptor Ativador de Fator Nuclear kappa-B , Receptores Citoplasmáticos e Nucleares/metabolismo , Receptores do Fator de Necrose Tumoral , Fator de Crescimento Transformador beta/antagonistas & inibidores , Fator de Crescimento Transformador beta/imunologiaRESUMO
A number of bone diseases characterised by excessive osteolysis (e.g. osteoporosis and Paget's disease) exhibit a marked gender difference in prevalence and are more common in the elderly population. Bone resorption is carried out by osteoclasts, which are formed by fusion of circulating mononuclear precursor cells of haematopoietic origin. In this study, we have determined whether there are gender- and age-related differences in osteoclast formation from circulating precursors. Peripheral blood mononuclear cells (PBMCs) were co-cultured with UMR106 osteoblast-like cells in the presence of macrophage-colony stimulating factor (M-CSF) and 1,25 dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) or cultured alone in the presence of sRANKL (soluble receptor activator of nuclear factor kappa B ligand) and M-CSF. As assessed by the formation of tartrate resistant acid phosphatase (TRAP)-positive (TRAP(+)) and vitronectin receptor-positive (VNR(+)) multinucleated cells (MNCs), there was no difference in the number of circulating osteoclast precursors in males and females. Lacunar resorption carried out by osteoclasts formed from these precursors was generally increased in males compared with females (P=0.03). An increase in the number of TRAP(+) and VNR(+) MNCs formed from male PBMCs was noted in response to 1,25(OH)(2)D(3) (P<0.005). An increase in lacunar resorption in cultures of PBMCs (10(5) per well) from males was also noted in response to 10(-9) M 1,25(OH)(2)D(3) (P<0.05) and sRANKL (P=0.05), but not M-CSF. The addition of dexamethasone resulted in a marked increase in osteoclast formation and lacunar resorption in both males and females. Post-menopausal females and males of comparable age showed similar levels of osteoclastogenesis. Pre-menopausal women showed similar levels of osteoclastogenesis but less resorption (P=0.01) compared with males of comparable age. These results show that there are specific gender/age-related differences in osteoclast formation and bone resorption and have implications for evaluating osteoclastogenesis in skeletal diseases such as primary osteoporosis and Paget's disease.
Assuntos
Proteínas de Bactérias , Osteoclastos/citologia , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/análise , Reabsorção Óssea , Calcitriol/farmacologia , Proteínas de Transporte/farmacologia , Diferenciação Celular/fisiologia , Células Cultivadas , Dexametasona/farmacologia , Feminino , Glucocorticoides/farmacologia , Humanos , Fator Estimulador de Colônias de Macrófagos/farmacologia , Masculino , Glicoproteínas de Membrana/farmacologia , Pessoa de Meia-Idade , Osteoclastos/química , Osteoclastos/metabolismo , Osteoporose/metabolismo , Osteoporose/patologia , Ligante RANK , Proteínas de Ligação a RNA/análise , Proteínas de Ligação a RNA/metabolismo , Receptor Ativador de Fator Nuclear kappa-B , Receptores de Vitronectina/análise , Receptores de Vitronectina/metabolismo , Fatores Sexuais , Fatores de Transcrição/análise , Fatores de Transcrição/metabolismoRESUMO
Using full-spectrum near infrared spectroscopy (fsNIRS), we measured changes in oxy- and deoxyhemoglobin (HbO2 and Hb), total hemoglobin (T-Hb) concentration, and hemoglobin oxygen saturation (SbO2) in the brain tissue of seven neonates immediately following birth. It was found that HbO2 rose rapidly within 2-3 min after birth. During the same time, there was a transient increase in T-Hb concentration, after which it decreased together with Hb. SbO2 increased rapidly after birth, from 18% at 1.5 min to about 55% at 5-6 min, followed by a gradual increase of about 10%. Oxygenation in the brain occurred much sooner in three subjects given oxygen for a short time immediately after birth than in those who did not receive oxygen. This preliminary study indicated that dynamic changes occur in cerebral circulation and oxygenation as part of the physiological changes taking place soon after birth.
Assuntos
Encéfalo/metabolismo , Hemoglobina Fetal/metabolismo , Consumo de Oxigênio , Espectroscopia de Luz Próxima ao Infravermelho , Hemoglobinas/metabolismo , Humanos , Recém-Nascido , Oxiemoglobinas/metabolismoRESUMO
We previously reported that PLA(2) activity in the gills is higher than that in other tissues in red sea bream and purified PLA(2) from the gills belongs to the group IB PLA(2) as well as other red sea bream PLA(2)s. In this study, we reconfirmed that the level of PLA(2) activity is extremely high in the gills compared with other tissues, and gill PLA(2) was detected only in the gills by immunoblotting and inhibition test using anti-gill PLA(2) monoclonal antibody. The level of PLA(2) activity and protein expression in the gills are well correlated. Fish can be roughly divided into high and low groups based on the level of PLA(2) activity. Gill PLA(2) was detected in the gills of the high group, but not the low group by immunoblotting. In the gills of the high group, gill PLA(2) was detected in the mucous cells and pavement cells located on the surface of gill epithelia by immunohistochemistry. On the other hand, positive signals were observed only in the mucous cells by in situ hybridization. We also isolated inactive proPLA(2), having AR propeptide, preceding the mature enzyme from the gill extract. These results suggest that gill PLA(2) is synthesized as an inactive proPLA(2) in the mucous cells and is secreted to the surface of gill epithelia.
Assuntos
Brânquias/enzimologia , Fosfolipases A/isolamento & purificação , Dourada , Animais , Anticorpos Monoclonais/imunologia , Cromatografia Líquida de Alta Pressão , Imuno-Histoquímica , Isoenzimas/análise , Isoenzimas/imunologia , Isoenzimas/isolamento & purificação , Camundongos , Camundongos Endogâmicos BALB C , Especificidade de Órgãos , Fosfolipases A/análise , Fosfolipases A/imunologiaRESUMO
Phospholipase A2 (PLA2) activity was investigated in various tissues of male and female red sea bream. In both male and female fishes, the specific activity of PLA2 in the gills was 70 times higher than that in other tissues, such as the adipose tissue, intestine, and hepatopancreas. Therefore, we tried to purify PLA2 from the gill filaments of red sea bream to near homogeneity by sequential chromatography on Q-Sepharose Fast Flow, Butyl-Cellulofine, and DEAE-Sepharose Fast Flow columns, and by reversed-phase high-performance liquid chromatography. Two minor and one major PLA2, tentatively named G-1, G-2 and G-3 PLA2, were purified, and all showed a single band with an apparent molecular mass of approximately 15 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis. The exact molecular mass values of G-1, G-2, and G-3 PLA2 were 14,040, 14,040 and 14,005 Da, respectively. G-1, G-2, and G-3 PLA2 had a Cys 11 and were all identical in N-terminal amino acid sequences from Ala-1 to Glu-56. A full-length cDNA encoding G-3 PLA2 was cloned by reverse transcriptase-polymerase chain reaction and rapid amplification of cDNA ends methods, and G-3 PLA2 was found to be classified to group IB PLA2 from the deduced amino acid sequence. G-1, G-2, and G-3 PLA2 had a pH optimum in an alkaline region at around pH 9-10 and required Ca2+ essentially for enzyme activity, using a mixed-micellar phosphatidylcholine substrate with sodium cholate. These results demonstrate that three group I PLA2, G-1, G-2, and G-3 PLA2, are expressed in the gill filaments of red sea bream.
Assuntos
Peixes/genética , Fosfolipases A/química , Fosfolipases A/genética , Tecido Adiposo/enzimologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Cálcio/metabolismo , Cromatografia em Agarose , Cromatografia Líquida de Alta Pressão , Clonagem Molecular , DNA Complementar/metabolismo , Soluções para Diálise/metabolismo , Sistema Digestório/enzimologia , Venenos Elapídicos/farmacologia , Eletroforese em Gel de Poliacrilamida , Feminino , Brânquias/enzimologia , Concentração de Íons de Hidrogênio , Intestinos/enzimologia , Masculino , Micelas , Dados de Sequência Molecular , Pâncreas/enzimologia , Fosfolipases A/metabolismo , Fosfolipases A2 , Reação em Cadeia da Polimerase , Isoformas de Proteínas , RNA Mensageiro/metabolismo , Análise de Sequência de Proteína , Homologia de Sequência de Aminoácidos , Colato de Sódio/farmacologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Suínos , Fatores de Tempo , Distribuição TecidualRESUMO
Two cDNA encoding red sea bream DE-1 and DE-2 phospholipases A2 (PLA2) were cloned from the hepatopancreas of red sea bream, Pagrus (Chrysophrys) major. The cDNA of DE-1 PLA2 encoded a mature protein of 125 amino acid residues with an apparent signal peptide of 20 residues and propeptide of 5 residues, and that of DE-2 PLA2, a mature protein of 126 amino acid residues with an apparent signal peptide of 17 residues and propeptide of 6 residues. Comparison of the predicted amino acid sequences for mature DE-1 and DE-2 PLA2 showed that both proteins contain 14 cysteines including Cys 11 and 77 and a pancreatic loop, which are commonly conserved in group IB PLA2; however, the identity in amino acid sequence between DE-1 and DE-2 PLA2 was low (47%). A previous report concerning the cDNA cloning of red sea bream gill G-3 PLA2 and the present results represent the first cloning and sequencing of three distinct isoforms of group IB PLA2 in a single fish species, red sea bream. Reverse transcription-polymerase chain reaction analysis showed that DE-1 PLA2 mRNA was expressed in the hepatopancreas, pyloric ceca, intestine, spleen, gonad, stomach, and kidney, whereas gill G-3 PLA2 mRNA was expressed only in the gills and gonad. The expression of DE-2 PLA2 mRNA was detected in all of the tissues analyzed. These results indicate that three distinct isoforms of group IB PLA2, DE-1 and DE-2 PLA2 in hepatopanceas and gill G-3 PLA2, are expressed in a tissue-specific manner in red sea bream.