RESUMO
Resistance to antiemetic treatment with 5-hydroxytryptamine-3 receptor antagonist is an issue. This study evaluated the potential roles of ABCB1 and ABCG2 polymorphisms in antiemetic treatment resistance in patients with cancer previously enrolled in a randomized controlled trial. A total of 156 patients were evaluated for their responses to antiemetic therapy and then subdivided into granisetron or palonosetron groups. The genotypes were evaluated for their association with antiemetic efficacy in each treatment groups. Additional risk factors associated with complete response (CR) were examined using a multivariate regression analysis. No significant associations were identified for genetic polymorphisms in the palonosetron group. In the granisetron group, patients with ABCB1 2677TT and 3435TT genotypes had higher proportion of CR. In addition to ABCB1 polymorphisms, gender and cisplatin dose were associated with granisetron response by univariate analysis. Multivariate logistic regression analysis revealed that the ABCB1 3435C>T polymorphism and cisplatin dose were significant predictors of CR.
Assuntos
Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética , Antieméticos/uso terapêutico , Antineoplásicos/efeitos adversos , Cisplatino/efeitos adversos , Proteínas de Neoplasias/genética , Testes Farmacogenômicos , Polimorfismo de Nucleotídeo Único , Subfamília B de Transportador de Cassetes de Ligação de ATP/genética , Adulto , Idoso , Antieméticos/farmacocinética , Antineoplásicos/uso terapêutico , Cisplatino/uso terapêutico , Feminino , Genótipo , Granisetron/farmacocinética , Granisetron/uso terapêutico , Humanos , Isoquinolinas/farmacocinética , Isoquinolinas/uso terapêutico , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Palonossetrom , Quinuclidinas/farmacocinética , Quinuclidinas/uso terapêuticoRESUMO
Reported here is the first observation of the tunneling surface diffusion of a hydrogen (H) atom on water ice. Photostimulated desorption and resonance-enhanced multiphoton ionization methods were used to determine the diffusion rates at 10 K on amorphous solid water and polycrystalline ice. H-atom diffusion on polycrystalline ice was 2 orders of magnitude faster than that of deuterium atoms, indicating the occurrence of tunneling diffusion. Whether diffusion is by tunneling or thermal hopping also depends on the diffusion length of the atoms and the morphology of the surface. Our findings contribute to a better understanding of elementary physicochemical processes of hydrogen on cosmic ice dust.
RESUMO
Japanese paddy rice systems commonly adopt the rotation of vegetables, wheat and soybeans with paddy rice. Crop rotation may, however, increase the nutrient load in effluent discharged from the district because more fertilizer is applied to the rotation crops than is applied to paddy crops. We investigated a paddy-field district subject to collective crop rotation and quantified the annual nutrient load of effluent from the district in three consecutive years. The total annual exports of nitrogen and phosphorus over the investigation period ranged from 30.3 to 40.6 kg N ha(-1) and 2.62 to 3.13 kg P ha(-1). The results suggest that rotation cropping increases the effluent nutrient load because applied fertilizer is converted to nitrate, and surface runoff is increased due to the absence of shuttering boards at the field outlets.
Assuntos
Agricultura/métodos , Produtos Agrícolas/metabolismo , Nitrogênio/química , Fósforo/química , Poluentes Químicos da Água/química , Fertilizantes/análise , Japão , Nitrogênio/metabolismo , Oryza , Fósforo/metabolismo , Chuva , Glycine max , Triticum , Verduras , Poluentes Químicos da Água/metabolismoRESUMO
BACKGROUND: Eosinophils and nasal polyps are believed to affect the surgical outcome of chronic rhinosinusitis (CRS). CRS is classified based on the presence of nasal polyps in western countries. The majority of patients with CRS with nasal polyps (CRS with NP) are characterized by predominantly eosinophilic inflammation. However, Asian patients with CRS with NP show characteristics indicative of neutrophilic inflammation. Therefore, are eosinophils or nasal polyps more important for the classification of CRS? METHODS: A prospective cohort study conducted from April 2007 to March 2008 classified patients with CRS based on the presence of nasal polyps and mucosal eosinophilia. The recurrence rate of nasal polyps was compared between the groups. Recurrence rate was analysed as a time-dependent variable by the Kaplan-Meier method. RESULTS: Eosinophilic inflammation was found in 59.6% of patients with CRS with NP. Patients with mucosal eosinophilia had higher polyp recurrence rate than patients without mucosal eosinophilia, whereas patients with nasal polyps did not have higher polyp recurrence rate than patients without nasal polyps. CONCLUSIONS: Presence of mucosal eosinophilia is a more important factor than nasal polyps for classifying CRS in terms of the surgical outcome.
Assuntos
Eosinofilia/epidemiologia , Rinite/classificação , Rinite/epidemiologia , Sinusite/classificação , Sinusite/epidemiologia , Adulto , Idoso , Doença Crônica , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mucosa Nasal , Pólipos Nasais/epidemiologia , Prognóstico , Estudos Prospectivos , RecidivaRESUMO
PURPOSE: To compare pupillary autonomic dysfunction in multiple system atrophy (MSA) and Parkinson's disease (PD). METHODS: We administered eye-drop tests to 40 MSA patients, 40 PD patients with similar disease duration, and 20 age-matched healthy controls. Pupillary supersensitivity to a parasympathomimetic agent (0.05% pilocarpine hydrochloride) and to a sympathomimetic agent (0.02% dipivefrine hydrochloride) was examined by assessing changes in pupil diameter. RESULTS: Pupillary supersensitivity to a parasympathomimetic agent (0.05% pilocarpine hydrochloride) and to a sympathomimetic agent (0.02% dipivefrine hydrochloride) was examined by assessing changes in pupil diameter. Pupillary supersensitivity to 0.05% pilocarpine was greatest among the PD patients (PD -23.1 +/- 14.4%, MSA -12.4 +/- 11.5%, control -9.5 +/- 8.2%, p < 0.05) but was not correlated with disease duration. Pupillary sensitivity to 0.02% dipivefrine was significantly greater in the PD and MSA patients versus controls (PD 10.5 +/- 12.0%, MSA 11.8 +/- 11.0%, control 3.1 +/- 5.8%, p < 0.05). MSA patients had pupillary sympathetic dysfunction from an early stage, whereas in PD patients it tended to gradually accelerate as the disease advanced. In MSA patients, pupillary sympathetic sensitivity to 0.02% dipivefrine was correlated with the severity of orthostatic hypotension during a head-up tilt test and with the elevation of systolic blood pressure during a noradrenaline infusion test. In PD patients, pupillary sympathetic sensitivity to 0.02% dipivefrine was correlated with a reduction of the heart-to-mediastinum (H/M) ratio using delayed-phase iodine-123 meta-iodobenzylguanidine ((123)I-MIBG) myocardial scintigraphy. CONCLUSION: These data indicate that eye-drop tests can reveal differences in the progression of pupillary autonomic dysfunction in patients with MSA and PD.
Assuntos
Doenças do Sistema Nervoso Autônomo/diagnóstico , Doenças do Sistema Nervoso Autônomo/etiologia , Atrofia de Múltiplos Sistemas/complicações , Atrofia de Múltiplos Sistemas/diagnóstico , Doença de Parkinson/complicações , Reflexo Pupilar/fisiologia , 3-Iodobenzilguanidina , Idoso , Sistema Nervoso Autônomo/fisiologia , Doenças do Sistema Nervoso Autônomo/diagnóstico por imagem , Pressão Sanguínea/fisiologia , Progressão da Doença , Feminino , Testes de Função Cardíaca , Humanos , Masculino , Pessoa de Meia-Idade , Mióticos , Atrofia de Múltiplos Sistemas/diagnóstico por imagem , Doença de Parkinson/diagnóstico por imagem , Doença de Parkinson/fisiopatologia , Pilocarpina , Pupila/fisiologia , Curva ROC , Cintilografia , Compostos RadiofarmacêuticosRESUMO
Electroluminescence (EL) devices for printable electronics using coprecipitated ZnS:Mn nanocrystal (NC) ink are demonstrated. The EL properties of these devices were investigated along with the structural and optical properties of ZnS:Mn NCs with an emphasis on their dependence on crystal size. Transmission electron microscopy and x-ray diffraction studies revealed that the NCs, with a crystal size of 3-4 nm, are nearly monodisperse; the crystal size can be controlled by the Zn(2+) concentration in the starting solution for coprecipitation. The results of optical studies indicate the presence of quantum confinement effects; in addition, the NC surfaces are well passivated, regardless of the crystal size. Finally, an increase in the luminance of EL devices with a decrease in crystal size is observed, which suggests the excitation mechanism of ZnS:Mn NC EL devices.
Assuntos
Eletrônica/instrumentação , Tinta , Medições Luminescentes/instrumentação , Manganês/química , Nanoestruturas/química , Impressão/instrumentação , Compostos de Selênio/química , Compostos de Zinco/química , Cristalização/métodos , Desenho de Equipamento , Análise de Falha de Equipamento , Precipitação Fracionada , Teste de Materiais , Nanoestruturas/ultraestrutura , Nanotecnologia/instrumentação , Tamanho da PartículaRESUMO
BACKGROUND: The incidence of aberrant bile duct injury associated with laparoscopic cholecystectomy (LC) has not yet been adequately examined. This study aimed to clarify the types of normal cystic ducts and the incidence of aberrant extrahepatic bile ducts, and to search for a method of avoiding injuries during LC. METHODS: Aberrant hepatic ducts were retrospectively categorized into five types according to the pattern of the cystic ducts and the accessory hepatic ducts by preoperative endoscopic retrograde cholangiography or multidetector three-dimensional computed tomography using drip infusion cholangiography. The aberrant bile ducts were classified as type A (merging at the right side of the common bile duct), type B (merging at the anterior side), or type C (merging at the posterior left side). RESULTS: The intrahepatic bile ducts and cystic duct were clearly shown for 1,044 of the 1,278 patients who underwent LC. Secondary branches of aberrant cystic ducts were observed in 37 cases (3.5%), and accessory hepatic ducts were observed in 30 cases (2.9%). A comparison of the difficulties encountered with LC for each type based on the merging patterns of cystic ducts showed that type C needed a much longer operation time for LC than the other types. CONCLUSIONS: A preoperative evaluation of the bile duct tract and the accessory hepatic duct before LC is important. Patients with a cystic duct merging normally into the posterior left side of the common hepatic duct (type C) experienced difficulty when undergoing LC. The authors have safely performed LC with the use of an endoscopic nasobiliary drainage tube in type D cases (cystic duct merging with the right hepatic duct), in type IV cases (cystic duct merging with an accessory hepatic duct).
Assuntos
Ductos Biliares Extra-Hepáticos/anatomia & histologia , Colecistectomia Laparoscópica , Cuidados Pré-Operatórios , Ductos Biliares Extra-Hepáticos/anormalidades , Ductos Biliares Extra-Hepáticos/lesões , Feminino , Humanos , Complicações Intraoperatórias/epidemiologia , Complicações Intraoperatórias/prevenção & controle , Masculino , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
BACKGROUND: Some evidence suggests that females have a lower pain threshold and a lower tolerance to painful stimuli. This study investigated gender differences in postoperative pain after laparoscopic cholecystectomy (LC) on the basis of visual analog pain scale (VAS) scores and the clinical course. METHODS: The 100 patients in this study (46 males and 54 females) underwent LC for cholecystolithiasis or gallbladder polyps without intraoperative complications. An 8-mm Penrose drain was retained for 42 h below the liver bed. All the patients were hospitalized for 4 days after LC, and the pain reported by patients, the time course of changes in the highest body temperature, the leukocyte count, and the C-reactive protein level were studied comparatively for the male and female patients. RESULTS: The VAS scores were significantly higher for the female patients than for the male patients at 24 h (62.7 +/- 24.6 vs 47.0 +/- 23.3; p = 0.0015) and at 48 h (39.2 +/- 24.3 vs 28.3 +/- 19.1; p = 0.0137) after LC. The female patients used analgesics more frequently and had significantly higher body temperatures than the male patients on day 1 (37.2 +/- 0.6 vs 36.9 +/- 0.4; p = 0.0037) and day 2 (36.9 +/- 0.6 vs 36.6 +/- 0.4; p = 0.0037) after surgery. CONCLUSIONS: Early postoperative pain after LC was more severe in female patients, and patients with high VAS scores tended to use analgesics more frequently.
Assuntos
Colecistectomia Laparoscópica , Dor Pós-Operatória/epidemiologia , Adulto , Idoso , Proteína C-Reativa/análise , Colecistolitíase/cirurgia , Feminino , Doenças da Vesícula Biliar/cirurgia , Humanos , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Medição da Dor , Limiar da Dor , Pólipos/cirurgia , Fatores SexuaisRESUMO
beta-Citryl-L-glutamate-hydrolysing enzyme (beta-CGHE) was purified from rat testis particulate fraction 13,000-fold, at a yield of 7%. The enzyme was purified by ammonium sulfate fractionation, hydroxyapatite, chelating Sepharose, beta-CG-Sepharose affinity chromatography and Sephacryl S-300 gel filtration. The purified enzyme usually migrated as two periodic acid Schiff's-stained bands on native polyacrylamide gel-electrophoresis (PAGE) with molecular weights of 350 and 420 kDa. Both bands hydrolyzed beta-citryl-L-glutamate (beta-CG) to citrate and glutamate. The 420 kDa band was changed by digestion with N-glycosidase F, into a 350 kDa band on native PAGE. The purified enzyme was composed of 90, 100, 115 and 130 kDa subunits on SDS-PAGE under non-reduced conditions. The purified enzyme was pharmacologically similar to the beta-CGHE activity partially purified from rat testis. This enzyme required manganese ions for full activity and it was strongly inhibited by nucleotides such as ATP or GTP and phosphate ions. beta-CGHE was also potently inhibited by an excitatory amino acid agonist, L-quisqualate, but not by another agonists, N-methyl-D-aspartate and kinate. It had high substrate specificity for beta-CG. The antibodies against the purified enzyme reacted mainly to the 115 kDa band on the SDS-PAGE and precipitated the enzyme activity from the crude and purified enzyme solution.
Assuntos
Amidoidrolases/isolamento & purificação , Testículo/enzimologia , Amidoidrolases/antagonistas & inibidores , Amidoidrolases/química , Animais , Cátions Bivalentes , Soros Imunes/química , Masculino , Peso Molecular , Nucleotídeos/farmacologia , Testes de Precipitina , Ácido Quisquálico/farmacologia , Ratos , Ratos Wistar , Especificidade por SubstratoRESUMO
To understand the calcium handling in whole heart having automaticity of the sinus node, we have developed a system of in situ imaging the intracellular calcium ion concentration in the perfused whole heart of the rat. The system consists of a stage-fixed upright microscope equipped with a real-time confocal laser scanning device of a multipinhole type with a water-immersion objective lens for observation. This in situ imaging system rendered observations and analyses of the rapidly changing images of intracellular calcium dynamics possible in the whole rat heart loaded with fluo-3. The scanning was conducted at a video rate of 30 frames per second, and the confocal effects included both X and Y planes. Calcium waves were frequently interrupted by calcium transients from either external electro-stimulation pulses or spontaneous sinus rhythm. Our findings suggest that abnormal calcium waves in minute areas cannot disturb the excitation-contraction coupling in the whole heart if the myocardial cells have orderly end-on-end intercellular electric paths.
Assuntos
Cálcio/metabolismo , Coração/fisiologia , Microscopia Confocal/métodos , Animais , Estimulação Elétrica , Processamento de Imagem Assistida por Computador , Masculino , Perfusão , Ratos , Ratos WistarRESUMO
Most of Earth's diamonds are connected with deep-seated mantle rocks; however, in recent years, µm-sized diamonds have been found in shallower metamorphic rocks, and the process of shallow-seated diamond formation has become a hotly debated topic. Nanodiamonds occur mainly in chondrite meteorites associated with organic matter and water. They can be synthesized in the stability field of graphite from organic compounds under hydrothermal conditions. Similar physicochemical conditions occur in serpentinite-hosted hydrothermal systems. Herein, we report the first finding of nanodiamonds, primarily of 6 and 10 nm, in Hyblean asphaltene-bearing serpentinite xenoliths (Sicily, Italy). The discovery was made by electron microscopy observations coupled with Raman spectroscopy analyses. The finding reveals new aspects of carbon speciation and diamond formation in shallow crustal settings. Nanodiamonds can grow during the hydrothermal alteration of ultramafic rocks, as well as during the lithogenesis of sediments bearing organic matter.
RESUMO
The nerve growth factor-sensitive phosphoprotein from PC12 cells, previously designated Nsp100, has been shown to be elongation factor 2 (EF-2). The criteria used for this identification include: (i) similarity of N-terminal sequence; (ii) phosphorylation by the same kinase; (iii) ADP-ribosylation mediated by diphtheria toxin; (iv) comparable function in cell-free protein synthesis. According to these criteria, Nsp100 and EF-2 are identical and the kinase that phosphorylates Nsp100 in PC12 cells is calcium/calmodulin kinase III.
Assuntos
Proteínas do Tecido Nervoso/análise , Fatores de Alongamento de Peptídeos/análise , Adenosina Difosfato Ribose/metabolismo , Sequência de Aminoácidos , Animais , Dados de Sequência Molecular , Fator 2 de Elongação de Peptídeos , Feocromocitoma , Fosfoproteínas/análise , Fosforilação , Ratos , Células Tumorais CultivadasRESUMO
To study the structure-activity relationships of brain injury-derived neurotrophic peptide (BINP), 12 analogs were synthesized by replacing each amino acid residue with Gly. BINP showed CD spectra typical of an alpha-helical conformation in TFE solution which mimics the membrane environment. In the alpha-helical conformation, BINP showed an amphiphilic profile. Neurotrophic activities of BINP and its analogs were estimated from the effects on supporting septal cholinergic neurons and on rescuing hippocampal neurons from injury caused by glutamate. Both assays showed that the residues on the hydrophobic side of the amphiphilic helix were essential for the neurotrophic activity.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/química , Fator Neurotrófico Derivado do Encéfalo/farmacologia , Proteínas do Tecido Nervoso/química , Proteínas do Tecido Nervoso/farmacologia , Neurônios/efeitos dos fármacos , Estrutura Secundária de Proteína , Sequência de Aminoácidos , Animais , Lesões Encefálicas/metabolismo , Fator Neurotrófico Derivado do Encéfalo/análogos & derivados , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Dicroísmo Circular , Ácido Glutâmico/farmacologia , Hipocampo/citologia , Hipocampo/efeitos dos fármacos , Neurônios/citologia , Sistema Nervoso Parassimpático/citologia , Sistema Nervoso Parassimpático/efeitos dos fármacos , RatosRESUMO
A cell-free extract of the anaerobic eubacterium, Clostridium thermoaceticum, catalyzes the synthesis of 5-aminolevulinic acid (ALA) from glutamate via the C5 pathway. The enzyme reaction resembles that of higher plants and algae in cofactor requirements and sensitivity to ribonuclease. From the phylogenetic distribution it is proposed that the C5 pathway evolved earlier than the ALA synthase pathway.
Assuntos
Ácido Aminolevulínico/biossíntese , Clostridium/metabolismo , Glutamatos/metabolismo , Ácidos Levulínicos/biossíntese , Carbono/metabolismo , Catálise , Sistema Livre de Células , Espectroscopia de Ressonância MagnéticaRESUMO
Biosynthesis of 5-aminolevulinic acid (ALA) in Chloroflexus aurantiacus, a thermophilic bacterium forming bacteriochlorophyll c, is shown to proceed via the C5 pathway by demonstrating (1) the specific labeling of its chlorin ring with [1 - 13C]glutamate and (2) the enzyme activity to produce ALA from glutamate in a cell-free extract. From the phylogenetic distribution it is suggested that ALA synthetase distributed in some aerobic eubacteria could be monophyletic in origin.
Assuntos
Ácido Aminolevulínico/metabolismo , Bactérias/metabolismo , Proteínas de Bactérias/metabolismo , Bacterioclorofilas/metabolismo , Bacterioclorofilas/química , Bacterioclorofilas/isolamento & purificação , Glutamatos/metabolismo , Espectroscopia de Ressonância Magnética/métodos , Estrutura MolecularRESUMO
Brain injury-derived neurotrophic peptide is a 13-amino acid peptide derived from a 15,000 mol. wt neurotrophic factor released from sites of mechanical injury in neonatal rat brain. This peptide promotes survival of septal cholinergic neurons and mesencephalic dopaminergic neurons, and protects hippocampal neurons from glutamate-induced neurotoxicity. In this study, we have developed a monoclonal antibody against a brain injury-derived neurotrophic peptide-binding protein by immunizing mice with septal synaptosomes from five-week-old rat brain. Monoclonal antibodies were screened for inhibition of the binding of a 125I-labeled analogue of brain injury-derived neurotrophic peptide to rat brain synaptosomes. The monoclonal antibody 6A22 suppressed the biological activity of brain injury-derived neurotrophic peptide and abolished the protective effect of the neurotrophic peptide against glutamate-induced neurotoxicity. This monoclonal antibody recognized a 40,000 mol. wt brain injury-derived neurotrophic peptide-binding protein, which was also identified by cross-linking experiments. Immunohistochemical studies showed that the 6A22 antibody bound to the cell surfaces of a subpopulation (about 60%) of hippocampal neurons in culture. These results are consistent with the possibility that the 40,000 mol. wt protein belongs to brain injury-derived neurotrophic peptide receptors.
Assuntos
Anticorpos Monoclonais/farmacologia , Proteínas do Tecido Nervoso , Neurônios/química , Animais , Morte Celular/efeitos dos fármacos , Morte Celular/fisiologia , Reagentes de Ligações Cruzadas/metabolismo , Meios de Cultura/farmacologia , Feto/citologia , Ácido Glutâmico/toxicidade , Hipocampo/citologia , Hibridomas , Radioisótopos do Iodo , Camundongos , Peso Molecular , Proteínas do Tecido Nervoso/análise , Proteínas do Tecido Nervoso/síntese química , Proteínas do Tecido Nervoso/imunologia , Neuroblastoma , Neurônios/citologia , Testes de Precipitina , Ligação Proteica/fisiologia , Ratos , Sinaptossomos/química , Células Tumorais CultivadasRESUMO
Brain injury-derived neurotrophic peptide is the fragmental 13-mer peptide of the novel neurotrophic factor which was extracted and purified from Sponge Gelform made of gelatin implanted at the mechanically-induced injury site in neonatal rat brains. Brain injury-derived neurotrophic peptide supports survival of septal cholinergic and mesencephalic dopaminergic neurons in culture, and rescues hippocampal neurons in culture from glutamate neurotoxicity. Here we studied the binding characteristics of brain injury-derived neurotrophic peptide to synaptosomes from normal adult rat brains and neurons in culture from neonatal rat brains. [125I]Asp-[Tyr11]-brain injury-derived neurotrophic peptide binding to rat brain synaptosomes was specific and saturable. Equilibrium binding studies revealed that [125I]Asp-[Tyr11]-brain injury-derived neurotrophic peptide bound to 1.1 pmol/mg protein with a Kd (dissociation constant) of 0.17 microM in hippocampal synaptosomes and to 2.0 pmol/mg protein with a Kd of 0.38 microM in septal synaptosomes. [125I]Asp-[Tyr11]-brain injury-derived neurotrophic peptide could bind to a subpopulation of hippocampal neurons in culture from embryonic rat brains. Affinity cross-linking with the carboxyl-reactive cross-linking reagent 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide-HCl and [125I]Asp-[Tyr11]-brain injury-derived neurotrophic peptide produced radiolabeled bands corresponding to 100,000, 50,000 and 40,000 mol. wt molecules on hippocampal neurons in culture. These results suggest that the 13-mer sequence of brain injury-derived neurotrophic peptide plays a crucial role in expressing the neurotrophic properties of the factor.
Assuntos
Proteínas do Tecido Nervoso/análise , Neurônios/química , Sinaptossomos/química , Animais , Autorradiografia , Sítios de Ligação/fisiologia , Morte Celular/fisiologia , Células Cultivadas , Reagentes de Ligações Cruzadas/metabolismo , Hipocampo/química , Hipocampo/citologia , Radioisótopos do Iodo , Proteínas do Tecido Nervoso/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Ratos , Núcleos Septais/química , Núcleos Septais/citologia , Sinaptossomos/metabolismoRESUMO
Interleukin-6 (human recombinant) supported the survival of cultured mesencephalic, catecholaminergic and septal cholinergic neurons from postnatal, two-week-old (P13-P15) rats. Significantly, more catecholaminergic neurons, stained by monoclonal anti-tyrosine hydroxylase antibody, were found in cultures supplemented with interleukin-6 at a concentration of 5 ng/ml than in cultures not treated with interleukin-6. The optimal dose used was 50 ng/ml. The survival effect of interleukin-6 on postnatal rat, tyrosine hydroxylase-positive neurons was observed both in cultures using serum-containing and serum-free medium. Contents of dopamine and noradrenaline in cultures with interleukin-6 were also larger than in control cultures. Interleukin-6 also increased the survival of cultured embryonic (E17) rat midbrain tyrosine hydroxylase-positive neurons. The effect on these neurons was, however, smaller, and the optimal dose of interleukin-6 was nearly 5 ng/ml. Interleukin-6 also supported the survival of cultured postnatal (P13) rat septal cholinergic neurons, visualized by acetylcholinesterase staining. The concomitant addition of mouse nerve growth factor (100 ng/ml) and interleukin-6 (50 ng/ml) had a synergetic effect on the survival of acetylcholinesterase-positive neurons in culture. Our data suggest that the survival of cultured tyrosine hydroxylase-positive, mesencephalic, and acetylcholinesterase-positive, septal neurons from postnatal two-week-old rats was supported by interleukin-6, just as there was a different dose dependency of interleukin-6 on the cultured postnatal neurons compared with embryonic neurons.
Assuntos
Catecolaminas/metabolismo , Interleucina-6/farmacologia , Mesencéfalo/metabolismo , Neurônios/efeitos dos fármacos , Sistema Nervoso Parassimpático/metabolismo , Septo Pelúcido/metabolismo , Animais , Animais Recém-Nascidos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Mesencéfalo/citologia , Neurônios/fisiologia , Ratos , Septo Pelúcido/citologiaRESUMO
The variable reactivity of OBF13 monoclonal antibody to mouse sperm was studied using a fluorescein activated cell sorter. Sperm from cauda epididymis were incubated with ionophore A23187, subjected to indirect immunostaining and analyzed by a cell sorter. Three peaks showing different fluorescence intensities were observed. These peaks contained (i) not stained (N), (ii) acrosomal cap or anterior region stained (A) and (iii) head stained (H) sperm, respectively. H type sperm showed more intense integral fluorescence than A type sperm. It was also noted that the H type were observed when cauda epididymal sperm were incubated with A23187, but not among non-incubated, or A23187 treated caput epididymal sperm. When sperm were pre-categorized as "dead" or "alive" by propidium iodide staining, no A type were observed in the "live" population. These results suggest that the sperm exhibiting the OBF13 antigen in the acrosome region lost their viability before they accomplished a "true" acrosome reaction.
Assuntos
Espermatozoides/fisiologia , Acrossomo/fisiologia , Animais , Anticorpos Monoclonais , Antígenos de Superfície/imunologia , Antígenos de Superfície/fisiologia , Calcimicina/farmacologia , Citometria de Fluxo , Imunofluorescência , Masculino , Camundongos , Camundongos Endogâmicos ICR , Camundongos Endogâmicos , Espermatozoides/imunologiaRESUMO
Dipeptidyl aminopeptidase, which hydrolyzes the 7-(Gly-Pro)-4-methylcoumarinamide, has been purified from the brains of 3 week-old rats. It was purified about 2,600-fold by column chromatography on CM-cellulose, hydroxyapatite and Gly-Pro AH-Sepharose. This enzyme hydrolyzed Lys-Ala-beta-naphthylamide well with an optimum pH of 5.5. It was inhibited by diisopropyl fluorophosphate, phenyl-methanesulfonyl fluoride, some cations, and puromycin, but was not inhibited by p-chloromercuribenzoate, N-ethylmaleimide, dithiothreitol, EDTA, iodoacetic acid, and bacitracin, indicating that rat brain dipeptidyl aminopeptidase is a serine protease. This enzyme showed a molecular weight of 220,000 by gel filtration and of 51,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The properties of purified rat brain dipeptidyl aminopeptidase were similar to those of bovine pituitary dipeptidyl peptidase II, but the molecular weight and substrate specificity of these enzymes were different.