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We report on precision spectroscopy of subwavelength confined molecular gases. This was obtained by rovibrational selective reflection of NH_{3} and SF_{6} gases using a quantum cascade laser at λ≈10.6 µm. Our technique probes molecules at micrometric distances (≈λ/2π) from the window of a macroscopic cell with submegahertz resolution, allowing molecule-surface interaction spectroscopy. We exploit the linearity and high resolution of our technique to gain novel spectroscopic information on the SF_{6} greenhouse gas, useful for enriching molecular databases. The natural extension of our work to thin cells will allow compact frequency references and improved measurements of the Casimir-Polder interaction with molecules.
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OBJECTIVE: Mesiodentes are usually found in the central position of the upper or lower jaw as supernumerary teeth. Here, we obtained 10 mesiodentes and three permanent teeth (PT) and separated the dental pulp (DP) from these into crown and root portions. We then characterized and compared the isolated crown portion-derived cells (crown cells) with root portion-derived cells (root cells) using a range of in vitro assays. MATERIALS AND METHODS: Crown cells and root cells were examined for cell surface marker expression, colony-forming unit-fibroblast (CFU-F), cell proliferation, cell cycle characteristics and markers, and osteogenic and adipogenic differentiation. RESULTS: The proportion of CD105-positive cells (CD105(+) cells) in the crown cells vs the root cells varied among the mesiodentes, but not among the PT. When there were more CD105(+) cells in the root cells than in the crown cells, the root cells showed higher CFU-F, proliferation capacity, and osteogenic differentiation capacity. In contrast, when the crown cells contained more CD105(+) cells than the root cells, the crown cells showed the higher CFU-F, proliferation capacity, and osteogenic differentiation capacity. In addition, the sorted CD105(+) cells showed higher CFU-F and proliferation capacity than the sorted CD105(-) cells. CONCLUSION: These results indicated that proportion of CD105(+) cells is an effective means of characterizing DP-derived cells in mesiodentes.
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Polpa Dentária/citologia , Células-Tronco Mesenquimais/citologia , Coroa do Dente/citologia , Raiz Dentária/citologia , Dente Supranumerário/patologia , Adolescente , Antígenos de Superfície/imunologia , Criança , Pré-Escolar , Ensaio de Unidades Formadoras de Colônias , Feminino , Citometria de Fluxo , Humanos , Masculino , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adulto JovemRESUMO
INTRODUCTION: Inadequate cerebral oxygen balance during cardiopulmonary bypass may cause neuropsychological dysfunction. Milrinone, a phosphodiesterase III inhibitor, augments cerebral blood flow by direct vasodilatation. We conducted a prospective, randomized study in patients undergoing cardiac surgery with cardiopulmonary bypass to clarify the clinical efficacy of milrinone in the imbalance of cerebral oxygen supply and demand during the rewarming period of cardiopulmonary bypass. METHODS: This is a prospective, randomized and placebo-controlled study. After anesthesia, a 5.5 F fiberoptic oximeter catheter was inserted into the right jugular bulb retrogradely for monitoring the jugular venous oxyhemoglobin saturation (SjO(2)). Patients were randomly assigned to two groups, one receiving a continuous infusion of milrinone, 0.5 µg/kg/min during hypothermic cardiopulmonary bypass, and the other receiving saline as control. RESULTS: Milrinone significantly prevented the reduction of the jugular venous oxyhemoglobin saturation at 10 minutes from the start of rewarming compared with the control group, but did not do so from 10 to 20 minutes after rewarming. CONCLUSION: Milrinone suppresses the reduction of SjO(2) and improves the balance of cerebral oxygen supply and demand during the early rewarming period of hypothermic cardiopulmonary bypass.
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Ponte Cardiopulmonar , Circulação Cerebrovascular/fisiologia , Hipotermia Induzida , Veias Jugulares/metabolismo , Milrinona/uso terapêutico , Oxigênio/metabolismo , Inibidores da Fosfodiesterase 3/uso terapêutico , Reaquecimento , Idoso , Circulação Cerebrovascular/efeitos dos fármacos , Transtornos Cognitivos/prevenção & controle , Feminino , Humanos , Infusões Intravenosas , Masculino , Pessoa de Meia-Idade , Milrinona/administração & dosagem , Milrinona/farmacologia , Oxiemoglobinas/metabolismo , Inibidores da Fosfodiesterase 3/administração & dosagem , Inibidores da Fosfodiesterase 3/farmacologia , Estudos Prospectivos , Fatores de TempoRESUMO
BACKGROUND: Edaravone, a free radical scavenger, has been shown to be neuroprotective in vivo and in vitro. However, the impact of small temperature variations on its neuroprotective actions remains unknown. METHODS: We examined the degree of neuroprotection conferred by various concentrations of edaravone on cortical cultures exposed to prolonged hypoxia (24 h) under three conditions: mild hypothermia (32 degrees C), normothermia (37 degrees C), and mild hyperthermia (39 degrees C). The survival of cortical neurones from E16 Wistar rats (SR) was evaluated using photomicrographs taken before and after exposure to hypoxia. RESULTS: The mean survival of neurones exposed to hypoxia at normothermia was 14.7 (sem 1.8)%. The addition of 50 microM edaravone significantly improved the mean survival to 40.5 (4.7)%. This improvement was noted at higher doses of edaravone (5 microM < or =) but not at lower doses (< or =500 nM). With mild hypothermia and prolonged hypoxia without edaravone, neuroprotection was significantly improved with a mean survival of 63.0 (5.2)%. This neuroprotective effect was not enhanced with the addition of edaravone, even at the highest dose. Hypoxia-induced neurotoxicity was aggravated by mild hyperthermia as reflected by a mean survival of 9.1 (2.1)%. However, higher concentrations of edaravone inhibited the deleterious effect of mild hyperthermia, thereby demonstrating a significant neuroprotective effect. The survival of neurones subjected to both hyperthermia and edaravone was the same as that of neurones exposed to normothermia and edaravone. CONCLUSIONS: Temperature is a potential factor in determining whether edaravone confers a neuroprotective effect when applied during prolonged hypoxic insults.
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Antipirina/análogos & derivados , Córtex Cerebral/efeitos dos fármacos , Sequestradores de Radicais Livres/farmacologia , Fármacos Neuroprotetores/farmacologia , Temperatura , Animais , Antipirina/farmacologia , Hipóxia Celular/fisiologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Córtex Cerebral/citologia , Relação Dose-Resposta a Droga , Edaravone , Neurônios/efeitos dos fármacos , Ratos , Ratos WistarRESUMO
Mouse susceptibility to experimental infections with flaviviruses is significantly influenced by a cluster of genes on chromosome 5 encoding a family of proteins with enzymatic properties, the 2'-5' oligoadenylate synthetases (OAS). Positional cloning of the locus in question has revealed that susceptibility of laboratory inbred strains to this class of virus is associated with a nonsense mutation in the gene encoding the OAS1B isoform. Analysis of the molecular structure of the cluster in different mammalian species including human indicates that the cluster is extremely polymorphic with a highly variable number of genes and pseudogenes whose functions are not yet completely established. Although still preliminary, a few recent observations also substantiate a possible role for OAS1 in human susceptibility to viral infections (West Nile virus, SARS, etc.) and its possible involvement in some other diseases such as type 1 diabetes and multiple sclerosis. Finally, convergent observations indicate that the molecules encoded by the 2 '-5' OAS cluster might be involved in other fundamental cellular functions such as cell growth and differentiation, gene regulation, and apoptosis.
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2',5'-Oligoadenilato Sintetase/fisiologia , Infecções por Flavivirus/imunologia , Imunidade Inata , 2',5'-Oligoadenilato Sintetase/genética , Animais , Infecções por Flavivirus/etiologia , Predisposição Genética para Doença , Humanos , CamundongosRESUMO
BACKGROUND: Hoarseness is a common complication after tracheal intubation and prolonged hoarseness may be very limiting for a patient. This study was designed to examine the duration of hoarseness after tracheal intubation and to identify risk factors that may increase the duration of hoarseness. METHODS: We prospectively studied 3093 adult patients (aged 18-77 yr), over a 3 yr period who required tracheal intubation. Postoperative hoarseness was assessed on the day of operation and on postoperative days 1, 3, and 7 by standardized interview by the resident anaesthetist managing the patient. If postoperative hoarseness was still present on postoperative day 7, the patient was followed up until complete resolution. We evaluated age, gender, weight, Cormack grades, duration of intubation, and the anaesthetic agents used as factors affecting the duration of hoarseness after tracheal intubation. RESULTS: Hoarseness was observed in 49% of patients on the day of surgery and in 29%, 11%, and 0.8% on 1, 3, and 7 postoperative days, respectively. Multiple regression analysis showed that patient age and duration of intubation, but not gender, weight, Cormack grades, or the agents used, were significant predictors of increased duration of hoarseness after tracheal intubation. We found three patients with arytenoid cartilage dislocation (0.097%) in our study population. CONCLUSIONS: The age of the patient and duration of intubation were significant factors in the duration of hoarseness after tracheal intubation. In addition, the incidence of arytenoid cartilage dislocation was 0.097%.
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Cartilagem Aritenoide/lesões , Rouquidão/etiologia , Intubação Intratraqueal/efeitos adversos , Adolescente , Adulto , Fatores Etários , Idoso , Feminino , Humanos , Luxações Articulares/etiologia , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias , Período Pós-Operatório , Estudos Prospectivos , Fatores de Risco , Fatores de Tempo , Adulto JovemRESUMO
BACKGROUND AND PURPOSE: We have previously demonstrated that stimulation of imidazoline receptors in the CNS prevented halothane-adrenaline arrhythmias during halothane anaesthesia and that stimulation of the vagus nerve may be critical to this effect. However, details of the mechanism(s) involved are not yet available. The present study was designed to examine the role of muscarinic receptors, protein kinase C (PKC), ATP-sensitive potassium channels (K(ATP)) and the mitochondrial permeability transition pore (MPTP) in the antiarrhythmic effect of rilmenidine, an imidazoline receptor agonist. EXPERIMENTAL APPROACH: Rats were anaesthetized with halothane and monitored continuously for arterial blood pressure and premature ventricular contractions. The arrhythmogenic dose of adrenaline was defined as the lowest dose producing three or more premature ventricular contractions within a 15-s period. We confirmed that centrally administered rilmenidine prevented halothane-adrenaline arrhythmias and then examined the antiarrhythmic effect of rilmenidine in the presence of atropine methylnitrate, a muscarinic receptor antagonist, calphostin C, a PKC inhibitor, HMR-1098, a sarcolemmal K(ATP) inhibitor, 5-hydroxydecanoic acid, a mitochondrial K(ATP) inhibitor or atractyloside, an MPTP opener. KEY RESULTS: The antiarrhythmic effect of rilmenidine was significantly inhibited by atropine methylnitrate, calphostin C, 5-hydroxydecanoic acid and atractyloside, but the effects of HMR-1098 in our model were not clear. CONCLUSIONS AND IMPLICATIONS: The present results suggest that muscarinic receptors, PKC, mitochondrial K(ATP) channels and MPTP may be crucial components of the mechanism involved in the antiarrhythmic effect of rilmenidine given into the CNS.
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Agonistas alfa-Adrenérgicos/farmacologia , Antiarrítmicos/farmacologia , Arritmias Cardíacas/prevenção & controle , Oxazóis/farmacologia , Animais , Arritmias Cardíacas/induzido quimicamente , Epinefrina , Canais KATP/efeitos dos fármacos , Canais KATP/metabolismo , Masculino , Proteínas de Transporte da Membrana Mitocondrial/efeitos dos fármacos , Proteínas de Transporte da Membrana Mitocondrial/metabolismo , Poro de Transição de Permeabilidade Mitocondrial , Proteína Quinase C/efeitos dos fármacos , Proteína Quinase C/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores Muscarínicos/efeitos dos fármacos , Receptores Muscarínicos/metabolismo , RilmenidinaRESUMO
OBJECTIVES: Following lesions in somatosensory pathways, deafferentation pain often occurs. Patients report that the pain is qualitatively complex, and its treatment can be difficult. Mirror visual feedback (MVF) treatment can improve deafferentation pain. We sought to classify the qualities of the pain in order to examine whether the potential analgesic effect of MVF depends on these qualities. METHODS: Twenty-two patients with phantom limb pain, or pain related to spinal cord or nerve injury, performed a single MVF procedure. Before and after the MVF procedure, we evaluated phantom limb awareness, movement representation of the phantom or affected/paralysed limb, pain intensity on an 11-point numerical rating scale (0-10) and the qualities of the pain [skin surface-mediated (superficial pain) vs deep tissue-mediated (deep pain)] using lists of pain descriptors for each of the two categories. RESULTS: Fifteen of the patients perceived the willed visuomotor imagery of the phantom or affected/paralysed limb after the MVF procedure. In most of the patients, a reduction in pain intensity and a decrease in the reporting of deep-pain descriptors were linked to the emergence of willed visuomotor imagery. CONCLUSIONS: In this pilot study, we roughly classified the pain descriptor items into two types for evaluating the qualities of deafferentation pain. We found that visually induced motor imagery by MVF was more effective for reducing deep pain than superficial pain. This suggests that the analgesic effect of MVF treatment does depend on the qualities of the pain. Further research will be required to confirm that this effect is a specific consequence of MVF.
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Biorretroalimentação Psicológica/métodos , Causalgia/terapia , Adolescente , Adulto , Idoso , Causalgia/etiologia , Feminino , Humanos , Imagens, Psicoterapia/métodos , Masculino , Pessoa de Meia-Idade , Medição da Dor/métodos , Membro Fantasma/terapia , Projetos Piloto , Desempenho Psicomotor , Resultado do TratamentoRESUMO
BACKGROUND: The dose of milrinone should be reduced in patients with renal failure. However, there is little data examining the relationship between plasma concentration of milrinone (pCmil) and renal function in intravenous infusion. METHODS: We evaluated the pCmil relative to renal function during intravenous infusion. We enrolled 10 heart failure patients. Milrinone was continuously infused at a rate of 0.2 microg/kg/min. Blood samples were collected at 6, 12, 24, and 48 h after the beginning of infusion. Urine was sampled during the first 24 h to calculate creatinine clearance (CLcr) and renal clearance of milrinone (rCLmil). RESULTS: The pCmil exhibited stability over 6 h after the beginning of infusion. During the first 24 h, CLcr and rCLmil were 62.2+/-30.6 ml/min and 1.67+/-0.77 ml/kg/min (106.2+/-60.3 ml/min), respectively. The rCLmil was highly correlated with CLcr. Y=1.77X-3.89 (X, CLcr; Y, rCLmil; R(2)=0.809, P<0.0001). Significant correlations were observed between CLcr and the plasma concentration during the continuous infusion. This correlation was expressed as the equation Y=51.1 x (BW/X)+28.2 (X; CLcr, Y; plasma concentration; BW, body weight; R(2)=0.695, P<0.01). CONCLUSION: The pCmil exhibited stability 6 h or later after the continuous infusion of milrinone 0.2 microg/kg/min. The pCmil can be estimated by the value of CLcr and BW.
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Cardiotônicos/sangue , Cardiopatias/complicações , Rim/efeitos dos fármacos , Milrinona/sangue , Insuficiência Renal/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Cardiotônicos/administração & dosagem , Cardiotônicos/urina , Creatinina/urina , Feminino , Humanos , Infusões Intravenosas , Testes de Função Renal , Masculino , Pessoa de Meia-Idade , Milrinona/administração & dosagem , Milrinona/urina , Fatores de TempoRESUMO
Two cold-sensitive transient receptor potential (TRP) channels, TRPA1 and TRPM8, have been identified and considered interesting because of their possible roles in thermosensation, nociception and other functions. Recently, we have reported that the phosphorylation of extracellular signal-regulated protein kinase and p38 mitogen-activated protein kinase occurred in primary afferent neurons in response to noxious heat stimulation of the peripheral tissue, i.e. activity-dependent activation of extracellular signal-regulated protein kinase and p38 in dorsal root ganglion neurons. In the present study, we investigated the phosphorylation of extracellular signal-regulated protein kinase, p38, and c-Jun N-terminal kinase in the rat dorsal root ganglion by cold stimulation using immunohistochemistry. Cold stimuli (28-4 degrees C) were applied by immersion of the hind paw into a water bath (six times of 10 s stimulation and 10 s interval, total 2 min). Noxious cold stimulation induced phosphorylated-extracellular signal-regulated protein kinase and phosphorylated-p38, but not phosphorylated-c-Jun N-terminal kinase, in small to medium diameter sensory neurons with a peak at 2 min after stimulation. We found that a cold stimulation at 4 degrees C showed a marked increase in the number of activated neurons. Furthermore, double staining for phosphorylated-extracellular signal-regulated protein kinase and phosphorylated-p38 showed no colocalization in the dorsal root ganglion neurons. We then performed double-labeling experiments for TRPA1 and TRPM8 mRNA and phosphorylation of mitogen-activated protein kinase. The majority of phosphorylated-extracellular signal-regulated protein kinase-positive neurons also expressed TRPM8 mRNA, whereas phosphorylated-p38 heavily colocalized with TRPA1 mRNA after noxious cold stimulation. Our data suggest that the noxious, but not innocuous, cold stimulation in vivo induced differential activation of extracellular signal-regulated protein kinase and p38 pathways in each subpopulation containing TRPA1 or TRPM8 in dorsal root ganglion.
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Canais de Cálcio/biossíntese , Temperatura Baixa , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Neurônios Aferentes/enzimologia , Medição da Dor/métodos , Canais de Cátion TRPM/biossíntese , Animais , Anquirinas , Temperatura Baixa/efeitos adversos , Ativação Enzimática/fisiologia , Indução Enzimática/fisiologia , Gânglios Espinais/enzimologia , Gânglios Espinais/metabolismo , Masculino , Proteínas Quinases Ativadas por Mitógeno/biossíntese , Neurônios Aferentes/metabolismo , Ratos , Ratos Sprague-Dawley , Canal de Cátion TRPA1 , Canais de Cátion TRPC/biossínteseRESUMO
Genomic aberrations at the chromosome 16q arm are one of the most consistent abnormalities observed by loss of heterozygosity and comparative genomic hybridization analyses in human prostate cancer, suggesting that there are tumor suppressor or metastasis suppressor genes encoded by this chromosomal region. To functionally identify such suppressor genes, we have conducted microcell-mediated chromosome transfer to introduce human chromosome 16 into the highly metastatic Dunning rat prostatic cancer cell line, AT6.1. The metastatic ability of the resultant microcell hybrid clones was then tested in a standard spontaneous metastasis assay using SCID mice. When the microcell-mediated chromosome transfer hybrid cells containing whole human chromosome 16 were injected, the number of metastatic lesions in the lung was significantly reduced as much as 99% on average. Therefore, chromosome 16 has a strong activity to suppress the metastatic ability of AT6.1 cells while it did not affect the tumorigenesis and tumor growth rate. A PCR analysis of various microcell hybrid clones with sequence-tagged site markers indicates that the metastasis suppressor activity is located in the q24.2 region of chromosome 16. Our results are consistent with the previous finding that the region of human chromosome 16q has frequent loss of heterozygosity in prostate cancer patients and suggest that there is a metastasis suppressor gene in this region that may play an important role in the progression of prostate cancer.
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Cromossomos Humanos Par 16 , Neoplasias Pulmonares/secundário , Neoplasias da Próstata/patologia , Animais , Caderinas/genética , Humanos , Perda de Heterozigosidade , Neoplasias Pulmonares/prevenção & controle , Masculino , Camundongos , Camundongos SCID , Neoplasias da Próstata/genética , Ratos , Transfecção , Células Tumorais CultivadasRESUMO
BACKGROUND: Decreased Gamma-aminobutyric acid (GABA)-ergic phasic inhibitory transmission in the spinal cord is thought to be responsible for the development of neuropathic pain. However, the role of GABAergic tonic current in substantia gelatinosa (SG) neurons in neuropathic pain remains to be fully elucidated. In this study, we assessed GABAergic tonic currents of SG neurons in a sciatic nerve chronic constriction injury (CCI) mouse. METHOD: Whole-cell patch clamp recordings form lumbar spinal cord slices was performed to evaluate GABAergic currents. We also investigated the expression changes of GABAA receptor subunits which are considered to mediate tonic currents. RESULTS: The percentage of SG neurons receiving GABAergic tonic currents decreased in CCI mice compared with Naïve mice. No significant change was observed in the mean amplitude of GABAergic tonic currents. RT-PCR and Western blot revealed that the expression of GABAA receptor δ subunits decreased following CCI. CONCLUSION: A reduction in the expression the δ subunit of the GABAA receptor and diminished GABAergic tonic current in SG neurons were observed after CCI in mice. GABAergic tonic current plays a key role in neuropathic pain. The GABAA receptor δ subunit may be a therapeutic target in neuropathic pain. WHAT DOES THIS STUDY ADD?: In spinal SG neurons, GABAergic inhibitory transmission operates through both phasic and tonic currents, but physiological role is largely unknown. In this study, we report dysregulation of GABAA receptor δ subunit-mediated tonic current in SG neurons may result in spinal disinhibition resulting in neuropathic pain in CCI mice.
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Neuralgia/metabolismo , Traumatismos dos Nervos Periféricos/metabolismo , Receptores de GABA-A/fisiologia , Nervo Isquiático/lesões , Substância Gelatinosa/fisiologia , Animais , Constrição , Modelos Animais de Doenças , Vértebras Lombares , Camundongos , Neuralgia/etiologia , Neuralgia/prevenção & controle , Neurônios/metabolismo , Técnicas de Patch-Clamp , Traumatismos dos Nervos Periféricos/etiologia , Ácido gama-Aminobutírico/metabolismoRESUMO
Materials at high pressures and temperatures are of great current interest for warm dense matter physics, planetary sciences, and inertial fusion energy research. Shock-compression equation-of-state data and optical reflectivities of the fluid dense oxide, Gd3Ga5O12 (GGG), were measured at extremely high pressures up to 2.6 TPa (26 Mbar) generated by high-power laser irradiation and magnetically-driven hypervelocity impacts. Above 0.75 TPa, the GGG Hugoniot data approach/reach a universal linear line of fluid metals, and the optical reflectivity most likely reaches a constant value indicating that GGG undergoes a crossover from fluid semiconductor to poor metal with minimum metallic conductivity (MMC). These results suggest that most fluid compounds, e.g., strong planetary oxides, reach a common state on the universal Hugoniot of fluid metals (UHFM) with MMC at sufficiently extreme pressures and temperatures. The systematic behaviors of warm dense fluid would be useful benchmarks for developing theoretical equation-of-state and transport models in the warm dense matter regime in determining computational predictions.
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We examined the effects of volatile anesthetics on the structure of the bacteriorhodopsin in the purple membrane by measurements of the absorption spectrum and the visible circular dichroism (CD) spectrum and assay of the retinal composition. As the concentrations of halothane, enflurane and methoxyflurane were increased, the absorption at 560 nm decreased but that at 480 nm increased with an isosbestic point around 510 nm. These anesthetic-induced spectroscopic changes were reversible. The CD spectrum showed the biphasic pattern with a positive and a negative band. As the concentration of halothane was increased from 4 mM to 8mM, the negative band reversibly diminished more drastically than the positive band, and at 8 mM of halothane the positive band shifted to around 480 nm. These results show that halothane disturbed the exciton coupling among bacteriorhodopsin molecules. The retinal isomer composition was analyzed using high performance liquid chromatography. The ratio of 13-cis- to all-trans-retinal was 47:53, 34:66 and 19:81 at control, 7.4 mM and 14.9 mM enflurane, respectively. After elimination of enflurane, the ratio returned to the control value. These findings indicate that volatile anesthetic directly affect a bacteriorhodopsin in the purple membrane and induce conformational changes in it.
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Anestésicos/farmacologia , Bacteriorodopsinas/metabolismo , Carotenoides/metabolismo , Proteínas de Membrana , Animais , Cromatografia Líquida de Alta Pressão , Dicroísmo Circular , Cricetinae , Escuridão , Enflurano/farmacologia , Halobacterium , Halotano/farmacologia , Isomerismo , Metoxiflurano/farmacologia , Conformação Proteica/efeitos dos fármacos , Retinaldeído/análise , EspectrofotometriaRESUMO
The rates of hydrolysis of p-nitrophenyl acetate (pNPA), p-nitrophenyl propionate (pNPP), p-nitrophenyl butanate (pNPB), and p-nitrophenyl valerate (pNPV) catalyzed by alpha-chymotrypsin (alpha-CHT) were measured with and without volatile anesthetics at 25.0 degrees C. Halothane activated the hydrolysis of pNPA and pNPP, meanwhile inhibited that of pNPB and pNPV. The activation phenomena were explained by the existence of a 1:1 enzyme-anesthetics complex and the opening of an activated pathway. The rate constant of pNPA hydrolysis catalyzed by alpha-CHT of the activated pathway kA by halothane was 0.269 s-1, whereas that of the normal pathway was k0 0.093 s-1. The free energy of activation was stabilized at 0.64 kcal/mol by halothane. The mechanisms of the activation and inhibition are discussed in terms of the molecular size of the substrate and anesthetics.
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Anestésicos/farmacologia , Quimotripsina/metabolismo , Animais , Sítios de Ligação , Bovinos , Enflurano/farmacologia , Ativação Enzimática , Halotano/farmacologia , Hidrólise , Isoflurano/farmacologia , Cinética , Matemática , Modelos Biológicos , Pâncreas/enzimologia , Especificidade por SubstratoRESUMO
We quantitatively assessed the spectroscopic changes of purple membrane in relation to the concentrations of a volatile anesthetic. As reported previously, volatile anesthetics show three modes of action on purple membrane. By using an anesthetic for which the concentration in solution could be determined spectroscopically and by applying modified analytical methods regarding the M-intermediate lifetime, we were able to clarify the quantitative relation between anesthetic concentration and each mode of action, a relation which in the past has only been described qualitatively. We also determined through the measurement of transient pH changes with pyranine that the proton pump efficiency per photochemical cycle in an action mode induced with low concentrations of anesthetic does not change from that of the native state. Moreover, we dynamically obtained the individual M-bacteriorhodopsin difference spectrum of each state at room temperature using our flash photolysis system equipped with a wavelength-tunable dye laser. These results demonstrated again that we should clearly distinguish different action modes of anesthetics according to their concentrations.
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Anestésicos/farmacologia , Membrana Purpúrea/efeitos dos fármacos , Bacteriorodopsinas/química , Clorofórmio/farmacologia , Relação Dose-Resposta a Droga , Concentração de Íons de Hidrogênio , Lasers , Metoxiflurano/farmacologia , Éteres Metílicos/farmacologia , Fotólise , Bombas de Próton/química , Membrana Purpúrea/química , Sevoflurano , Espectrofotometria , TemperaturaRESUMO
It has long been speculated that porcine cathelin is an N-terminal fragment of a longer precursor protein which possesses antimicrobial activity. In an attempt to find such a precursor, a cDNA clone was recently isolated and sequenced by screening a cDNA library from porcine bone marrow. In order to identify the functional activity of the putative protein encoded by an open reading frame, we have synthesized various lengths of peptides that correspond to the C-terminal region of the protein and examined them for their antimicrobial activities. We found that a 13 amino acid tryptophan-rich region with the sequence of VRRFPWWWPFLRR had strong antimicrobial activity with a wide spectrum. It showed potency against Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Staphylococcus epidermidis, Proteus mirabilis, and Streptococcus group D as well as Aspergillus fumigatus. The action of this peptide is bactericidal rather than bacteriostatic and this activity is completely inhibited by 2 mM MgCl2. Our results indicate that the previously identified putative precursor encoded by the isolated cDNA indeed possesses a potent antimicrobial activity and that this 13 amino acid synthetic peptide is considered to be a potentially effective drug against various infectious agents.
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Anti-Infecciosos , Bactérias/efeitos dos fármacos , Fragmentos de Peptídeos/farmacologia , Proteínas/química , Proteínas/metabolismo , Sequência de Aminoácidos , Animais , Antibacterianos , Aspergillus fumigatus/efeitos dos fármacos , Medula Óssea , Cálcio/farmacologia , Candida albicans/efeitos dos fármacos , DNA Complementar , Escherichia coli/efeitos dos fármacos , Biblioteca Gênica , Klebsiella pneumoniae/efeitos dos fármacos , Cloreto de Magnésio/farmacologia , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Fragmentos de Peptídeos/isolamento & purificação , Precursores de Proteínas/química , Precursores de Proteínas/metabolismo , Proteus mirabilis/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos dos fármacos , Staphylococcus epidermidis/efeitos dos fármacos , Streptococcus/efeitos dos fármacos , Relação Estrutura-Atividade , Suínos , TriptofanoRESUMO
We tested the ability of lithium (Li(+)) to block heat hyperalgesia, cold allodynia, mechanical allodynia and mechanical hyperalgesia in rats experimentally subjected to painful peripheral neuropathy. Chronic constrictive injury (CCI) to the sciatic nerve induced persistent hyperalgesia and allodynia. Intrathecal injection of Li(+) (2.5-40 micromol) into the region of lumbar enlargement dose-dependently reduced heat hyperalgesia, cold allodynia and mechanical allodynia for 2-6 h after injection, but had no effect on mechanical hyperalgesia. Li(+) had no significant effect on responses from control and sham-operated animals. Intrathecal injection of myo-inositol (2.5 mg) significantly reversed both the anti-hyperalgesic and anti-allodynic effect of Li(+). These findings suggest that intrathecal Li(+) suppresses neuropathic pain response in CCI rats through the intracellular phosphatidylinositol (PI) second messenger system in spinal cord neurons. Lithium (Li(+)) has already found widespread clinical application; these results suggest that its therapeutic utility may be extended to include treatment of neuropathic pain syndromes resulting from peripheral nerve injury.
Assuntos
Lítio/uso terapêutico , Dor/tratamento farmacológico , Doenças do Sistema Nervoso Periférico/complicações , Animais , Doença Crônica , Temperatura Baixa , Interações Medicamentosas , Temperatura Alta , Hiperalgesia/etiologia , Injeções Espinhais , Inositol/farmacologia , Lítio/administração & dosagem , Lítio/antagonistas & inibidores , Masculino , Dor/etiologia , Medição da Dor/efeitos dos fármacos , Estimulação Física , Ratos , Ratos Sprague-Dawley , Fatores de TempoRESUMO
1. We describe the effects of barbiturates on the neurotoxicity induced by nitric oxide (NO) on foetal rat cultured cortical and hippocampal neurones. Cessation of cerebral blood flow leads to an initiation of a neurotoxic cascade including NO and peroxynitrite. Barbiturates are often used to protect neurones against cerebrovascular disorders clinically. However, its neuroprotective mechanism remains unclear. 2. In the present experiment, we established a new in vitro model of brain injury mediated by NO with an NO-donor, 1-hydroxy-2-oxo-3-(3-aminopropyl)-3-isopropyl-1-triazene (NOC-5) on grid tissue culture wells. We also investigated the mechanisms of protection of CNS neurones from NO-induced neurotoxicity by thiopentone sodium, which contains a sulphydryl group (SH-) in the medium, and pentobarbitone sodium, which does not contain SH-. 3. Primary cultures of cortical and hippocampal neurones (prepared from 16-day gestational rat foetuses) were used after 13-14 days in culture. The cells were exposed to NOC-5 at the various concentrations for 24 h in the culture to evaluate a dose-dependent effect of NOC-5. 4. To evaluate the role of the barbiturates, neurones were exposed to 4, 40 and 400 microM of thiopentone sodium or pentobarbitone sodium with or without 30 microM NOC-5. In addition, superoxide dismutase (SOD) at 1000 u ml(-1) and 30 microM NOC-5 were co-administered for 24 h to evaluate the role of SOD. 5. Exposure to NOC-5 induced neural cell death in a dose-dependent manner in both cortical and hippocampal cultured neurones. Approximately 90% of the cultured neurones were killed by 100 microM NOC-5. 6. This NOC-5-induced neurotoxicity was significantly attenuated by high concentrations of thiopentone sodium (40 and 400 microM) as well as SOD, but not by pentobarbitone sodium. The survival rates of the cortical neurones and hippocampal neurones that were exposed to 30 microM NOC-5 were 11.2+/-4.2% and 37.2+/-3.0%, respectively, and in the presence of 400 microM thiopentone sodium, the survival rate increased to 65.3+/-3.5% in the cortical neurones and 74.6+/-2.2% in the hippocampal neurones. 7. These findings demonstrate that thiopentone sodium, which acts as a free radical scavenger, protects the CNS neurones against NO-mediated cytotoxicity in vitro. In conclusion, thiopentone sodium is one of the best of the currently available pharmacological agents for protection of neurones against intraoperative cerebral ischaemia.
Assuntos
Hipnóticos e Sedativos/farmacologia , Neurônios/efeitos dos fármacos , Óxido Nítrico/metabolismo , Pentobarbital/farmacologia , Tiopental/farmacologia , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Córtex Cerebral/citologia , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Sequestradores de Radicais Livres/farmacologia , Hipocampo/citologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Ratos , Ratos Wistar , Triazenos/toxicidadeRESUMO
OBJECTIVES: Transport ventilators with inspiratory triggering functions and pressure support-control modes have recently become commercially available. We evaluated these ventilators in comparison with a standard ICU ventilator. STUDY DESIGN: Laboratory study with a mechanical lung model. METHODS: We compared the performance of four transport ventilators (model 740, Mallinckrodt, Pleasanton, CA; TBird, Bird Products Corp, Palm Springs, CA; LTV1000, Pulmonetic Systems, Colton, CA; Esprit, Respironics, Vista, CA) with a standard ICU ventilator (model 7200ae; Mallinckrodt) using a test lung that simulated spontaneous breathing (compliance, 46.8 mL/cm H(2)O; resistance, 5 cm H(2)O/L/s). The settings of ventilators were positive end-expiratory pressure (PEEP) of 0 or 5 cm H(2)O, and pressure support (PS) of 0 or 10 cm H(2)O. The settings of the test lung were inspiratory time of 1 s, respiratory rate of 10/min, peak inspiratory flow of 40, 60, and 80 L/min. To evaluate inspiratory function at each setting, we measured the inspiratory delay time (DT), inspiratory trigger pressure (P-I), and the time for airway pressure to rise from the baseline pressure to 90% of the end-inspiratory pressure (T(90%)); for expiratory function, supraplateau expiratory pressure (P-E) and the time constant (taue) for pressure decrease during exhalation were evaluated. Oxygen requirement was assessed as the time required to empty a 3.5-L oxygen tank. RESULTS: For inspiratory triggering, four transport ventilators had DT < 100 ms, which is considered clinically satisfactory, in all the settings except for PS 0 cm H(2)O, PEEP 0 cm H(2)O, and inspiratory flow of 80 L/min with LTV1000. P-I increased only in LTV1000 when PEEP was increased from 0 to 5 cm H(2)O. taue for the transport ventilators was > 50% shorter than for the ICU ventilator except for PS 0 cm H(2)O and PEEP 5 cm H(2)O with TBird. Oxygen requirement was lowest for the Esprit, followed by the 740, LTV1000, and TBird. CONCLUSION: The newer Food and Drug Administration-approved transport ventilators have performance indexes comparable to the ventilator currently used in ICUs and can probably be recommended for clinical use.