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Convulsive status epilepticus (CSE) is a neurological disease with contraction and extension of limbs, leading to damage of hippocampus and cognition. This study aimed to explore the effects of dexmedetomidine (DEX) on the cognitive function and neuroinflammation in CSE rats. All rats were divided into control group, CSE group and DEX group. Morris water maze test was used to measure cognitive function. Acute hippocampal slices were made to detect long-term potentiation (LTP). Immunohistochemistry was used to determine the expression of α7-nicotinic acetylcholine receptor (α7-nAChR) and interleukin-1ß (IL-1ß). Enzyme-linked immunosorbent assay (ELISA) was used to measure serum levels of IL-1ß, tumor necrosis factor-α (TNF-α), S-100ß and brain-derived neurotrophic factor (BDNF). Our results showed that DEX improved the memory damage caused by CSE. DEX reduced seizure severity and increased the amplitudes and sustainable time of LTP, and also inhibited the hippocampal expression of α7-nAChR and IL-1ß in CSE rats. DEX treatment decreased serum IL-1ß, TNF-α and S-100ß levels and increased BDNF levels. The effects of DEX on seizure severity and LTP could be simulated by nicotine or attenuated by concurrent α-bungarotoxin (α-BGT) treatment. In conclusions, DEX significantly improved spatial cognitive dysfunction, reduced seizure severity and increased LTP in CSE rats. Improvements by DEX were closely related to enhancement of cholinergic anti-inflammatory pathway.
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Anti-Inflamatórios/uso terapêutico , Anticonvulsivantes/uso terapêutico , Dexmedetomidina/uso terapêutico , Convulsões/tratamento farmacológico , Estado Epiléptico/tratamento farmacológico , Animais , Fator Neurotrófico Derivado do Encéfalo/análise , Fator Neurotrófico Derivado do Encéfalo/imunologia , Hipocampo/efeitos dos fármacos , Hipocampo/imunologia , Hipocampo/patologia , Hipocampo/fisiopatologia , Interleucina-1beta/análise , Interleucina-1beta/imunologia , Potenciação de Longa Duração/efeitos dos fármacos , Masculino , Memória/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Convulsões/imunologia , Convulsões/patologia , Convulsões/fisiopatologia , Estado Epiléptico/imunologia , Estado Epiléptico/patologia , Estado Epiléptico/fisiopatologia , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/imunologia , Receptor Nicotínico de Acetilcolina alfa7/análise , Receptor Nicotínico de Acetilcolina alfa7/imunologiaRESUMO
In order to investigate the effect of benzothiadiazole (BTH) and ß-aminobutyric acid (BABA) on the resistance of tea plants (Camellia sinensis) to tea geometrid (Ectropis obliqua), three levels each of benzothiadiazole (BTH) and ß-aminobutyric acid (BABA) were sprayed on 10-year-old tea plants. Generally PPO and PAL activities increased with low concentrations of BTH and BABA treatments. Quantitative RT-PCR revealed a 1.43 and 2.72-fold increase in PPO gene expression, and 3.26 and 3.99-fold increase in PAL gene expression with 75 mg/L BTH and 400 mg/L BABA respectively. Analysis of hydrolysis of synthetic substrates also revealed that chymotrypsin-like enzyme activity present in larval midgut extracts was not significantly inhibited by BTH and BABA. However, proteinase activity was found to be inversely proportional to the age of tea geometrid. Larvae pupation rate decreased by 8.10, 10.81 and 21.62% when tea geometrid were fed with leaves treated with 25, 50 and 75 mg/L BTH solutions, while 100, 200 and 400 mg/L BABA solutions decreased same by 8.10, 16.21 and 13.51% respectively. Also, larvae development period delayed to 23.33 and 26.33 days with 75 mg/L BTH and 400 mg/L BABA treatments respectively. The results in this study; therefore, suggest that benzothiadiazole (BTH) and ß-aminobutyric acid (BABA) play a role in inducing resistance in tea plants to tea geometrid, with the optimal effect achieved at BTH-3 (75 mg/L) and BABA-3 (400 mg/L), respectively.
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Aminobutiratos/farmacologia , Camellia sinensis/crescimento & desenvolvimento , Resistência à Doença , Mariposas/efeitos dos fármacos , Tiadiazóis/farmacologia , Animais , Camellia sinensis/genética , Camellia sinensis/parasitologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Mariposas/crescimento & desenvolvimento , Doenças das Plantas/parasitologia , Doenças das Plantas/prevenção & controle , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/parasitologia , Proteínas de Plantas/genéticaRESUMO
Objective To investigate the value of 3D pseudo-continuous arterial spin labeling (3D-pCASL) magnetic resonance perfusion technique in evaluating posterior circulation ischemia (PCI) of the elderly beyond 80 years old and to offer the evidence of PCI of the elderly for clinical diagnosis. Methods Totally 53 male subjects older than 80 years were recruited in this study,including 20 subjects with clinically diagnosed PCI and 33 normal subjects. All the subjects underwent routine brain magnetic resonance imaging and 3D-pCASL sequence on a 3.0T magnetic resonance imaging system with 8 channel brain coil. Two post-labeling delay (PLD) time (PLD=1525 ms and PLD=2525 ms) of 3D-pCASL were used in this study to increase the accuracy of cerebral blood flow (CBF) change of posterior circulation region. We used SPM12 software to measure mean CBF values of bilateral occipital lobes and bilateral cerebellums. Independent sample t-test and rank-sum test were performed to evaluate the difference of CBF changes of anterior circulation and posterior circulation in two groups at two PLD time,the difference of CBF changes of bilateral occipital lobes and bilateral cerebellums in two groups of two PLD time,and the difference of increment of CBF between two PLD interval between two groups. Results In case group,the CBF value of the anterior circulation was significantly higher than that of posterior circulation at both two PLD time points (PLD=1525 ms and PLD=2525 ms)(P=0.000,P=0.000);in control group,the CBF value of the anterior circulation was significantly higher than that of the posterior circulation only at PLD=1525ms (P=0.025). The CBF values at bilateral occipital lobes and bilateral cerebellums at two PLD time points (PLD=1525 ms and PLD=2525 ms) were significantly higher in case group than in control group(P=0.003,P=0.002,P=0.000,P=0.001,P=0.000,P=0.001,P=0.002,P=0.014,respectively). Compared with the control group,the difference was statistically significant in bilateral occipital lobes and cerebellums with a smaller â³CBF between two PLD interval in case group (P=0.004,P=0.001,P=0.001,P=0.025). Conclusion Multiple PLD time points need to be used in 3D-pCASL in diagnosing PCI of the elderly because the posterior circulation is slow in these patients. 3D-pCASL technique is sensitive in detecting decreased CBF in posterior circulation and therefore can be used to predict posterior circulation stroke in the elderly.
Assuntos
Isquemia Encefálica/diagnóstico por imagem , Circulação Cerebrovascular , Angiografia por Ressonância Magnética , Imagem de Perfusão , Marcadores de Spin , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Humanos , Masculino , Acidente Vascular Cerebral/diagnóstico por imagemRESUMO
Leaf spreading is a key processing step that affects the aroma formation of green tea. The effects of a single-light wavelength on the aroma and taste of tea have been extensively studied. Less attention has been paid to the effect of different complex light intensities on the formation of green tea's volatile aroma during leaf spreading. The current study was designed to evaluate how leaf spreading under different complex light intensities relates to the quality of green tea. Using headspace solid-phase micro-extraction and gas chromatography-mass spectrometry (HS-SPME/GC-MS), volatile flavor compounds in green tea were analyzed during leaf spreading in five different light conditions. Multivariate statistical analysis and odor activity values (OAVs) were used to classify these samples and identify key odors. Eight distinct groups, including ninety volatile compounds, were detected. The most prevalent volatile compounds found in green tea samples were hydrocarbons and alcohols, which accounted for 29% and 22% of the total volatile compounds, respectively. Fourteen volatile compounds (OAV > 1) were identified as key active differential odorants. The chestnut-like aroma in green tea was mostly derived from 3-methyl-butanal and linalool, which were significantly accumulated in medium-intensity light (ML).
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Traditionally, fresh S. japonicum flowers (SJF) and S. japonicum flowers buds (SJFB) are dried prior to further processing and use. Here, we investigated the ways in which drying techniques, including sun drying (SD), steam drying (STD), microwave drying (MD), hot air drying (HAD, 40 °C, 60 °C, 80 °C, 100 °C), and freeze drying (FD), alter the flavonoid composition of freshly-harvested SJF and SJFB. The flavonoid content of dried samples was determined by Ultra High Performance Liquid Chromatography-Diode Array Detector (UPLC-DAD). Overall, different drying techniques had significantly different effects on the RU content, ranging from 10.63 % (HAD-80 °C) to 34.13 % (HAD-100 °C) in SJF and from 18.91 % (HAD-100 °C) to 29.16 % (HAD-40 °C) and 30.53 % (SD) in SJFB. To clarify the mechanism by which drying affects the RU content of S. japonicum flowers, we studied the activity of a rutin-hydrolyzing enzyme (RHE) isolated from SJF and SJFB using multiple separation and assay methods. According to the Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) results, the apparent molecular weight of the purified RHE was approximately 38 kDa. According to UPLC-DAD, RHE catalyzes the production of quercetin (QU) from rutin (RU), but not from other flavonoid glycosides. Drying fresh SJF and SJFB at low and high temperatures can inhibit RHE activity and prevent RU hydrolysis. Therefore, subjecting freshly-harvest SJF to HAD-100 °C, and freshly-harvest SJFB to SD or HAD-40 °C, can greatly increase the RU content. In particular, HAD is viable for large-scale application due to its simplicity and industrial feasibility.
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BACKGROUND: Visceral pain is common symptom involved in many gastrointestinal disorders such as inflammatory bowel disease. The underlying molecular mechanisms remain elusive. We investigated the molecular mechanisms and the role for voltage gated calcium channel (VGCC) in the pathogenesis in a rat model of 2,4,6-trinitrobenzenesulfonic acid (TNBS) induced visceral inflammatory hypersensitivity. RESULTS: Using Agilent cDNA arrays, we found 172 genes changed significantly in dorsal root ganglia (DRG) of TNBS treated rats. Among these changed genes, Cav1.2 and Cav2.3 were significantly up-regulated. Then the RT-PCR and Western blot further confirmed the up-regulation of Cav1.2 and Cav2.3. The whole cell patch clamp recording of acutely dissociated colonic specific DRG neurons showed that the peak IBa density was significantly increased in colonic neurons of TNBS treated rats compared with control rats (-127.82 ± 20.82 pA/pF Vs -91.67 ± 19.02 pA/pF, n = 9, *P < 0.05). To distinguish the different type of calcium currents with the corresponding selective channel blockers, we found that L-type (-38.56 ± 3.97 pA/pF Vs -25.75 ± 3.35 pA/pF, n = 9, * P < 0.05) and R-type (-13.31 ± 1.36 pA/pF Vs -8.60 ± 1.25 pA/pF, n = 9, * P < 0.05) calcium current density were significantly increased in colonic DRG neurons of TNBS treated rats compared with control rats. In addition, pharmacological blockade with L-type antagonist (nimodipine) and R-type antagonist (SNX-482) with intrathecal injection attenuates visceral pain in TNBS induced inflammatory visceral hypersensitivity. CONCLUSION: Cav1.2 and Cav2.3 in colonic primary sensory neurons play an important role in visceral inflammatory hyperalgesia, which maybe the potential therapeutic targets.
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Canais de Cálcio Tipo L/metabolismo , Canais de Cálcio Tipo R/metabolismo , Proteínas de Transporte de Cátions/metabolismo , Hipersensibilidade/patologia , Vísceras/patologia , Animais , Western Blotting , Canais de Cálcio Tipo L/genética , Canais de Cálcio Tipo R/genética , Carbocianinas/metabolismo , Proteínas de Transporte de Cátions/genética , Colo/efeitos dos fármacos , Colo/metabolismo , Colo/patologia , Gânglios Espinais/metabolismo , Gânglios Espinais/patologia , Perfilação da Expressão Gênica , Hipersensibilidade/genética , Injeções Espinhais , Masculino , Nimodipina/administração & dosagem , Nimodipina/farmacologia , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Venenos de Aranha/administração & dosagem , Venenos de Aranha/farmacologia , Ácido Trinitrobenzenossulfônico , Vísceras/efeitos dos fármacos , Vísceras/metabolismoRESUMO
Auxin affects all aspects of plant growth and development, including morphogenesis and adaptive responses. Auxin transmembrane transport is promoted by PIN formation (PIN) and a structurally similar PIN-like (PILS) gene family, which jointly controls the directional transport of the auxin between plant cells, and the accumulation of intracellular auxin. At present, there is no study investigating the roles of CslPIN and CslPILS gene family in root development in the tea plant (Camellia sinensis). In this study, 8 CslPIN and 10 CslPILS genes were identified in the tea plant, and their evolutionary relationships, physical and chemical properties, conserved motifs, cis-acting elements, chromosome location, collinearity, and expression characteristics were analyzed. The mechanism of CslPIN and CslPILS in the formation of tea adventitious roots (ARs) was studied by the AR induction system. Through functional verification, the regulation of CslPIN3 gene on root growth and development of tea plant was studied by over-expression of CslPIN3 in Arabidopsis thaliana and in situ hybridization in Camellia sinensis. The results confirmed CslPIN3 was involved in the regulation of root growth and development as well as auxin accumulation. This study provides a better insight into the regulatory mechanism of CslPIN and CslPILS gene family on the formation of AR in tea plant.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Camellia sinensis , Camellia sinensis/genética , Ácidos Indolacéticos/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Chá/metabolismo , Regulação da Expressão Gênica de Plantas , Raízes de Plantas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Folding, assembly, and trafficking of ion channels are tightly controlled processes and are important for biological functions relevant to health and disease. Here, we report that functional expression of the Eag channel is temperature-sensitive by a mechanism that is independent of trafficking or surface targeting of the channel protein. Eag channels in cells grown at 37 °C exhibit voltage-evoked gating charge movements but fail to conduct K(+) ions. By mutagenesis and chimeric channel studies, we show that the N- and C-terminal regions are involved in controlling a step after movement of the voltage sensor, as well as in regulating biophysical properties of the Eag channel. Synthesis and assembly of Eag at high temperature disrupt the ability of these domains to carry out their function. These results suggest an important role of the intracellular regions in the generation of Eag currents.
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Proteínas de Drosophila , Drosophila/fisiologia , Canais de Potássio Éter-A-Go-Go , Ativação do Canal Iônico/fisiologia , Potássio/metabolismo , Transporte Proteico/fisiologia , Sequência de Aminoácidos , Animais , Células CHO , Cricetinae , Cricetulus , Proteínas de Drosophila/química , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Canais de Potássio Éter-A-Go-Go/química , Canais de Potássio Éter-A-Go-Go/genética , Canais de Potássio Éter-A-Go-Go/metabolismo , Potenciais da Membrana/fisiologia , Proteínas de Membrana/química , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Dados de Sequência Molecular , Mutagênese , Dobramento de Proteína , Estrutura Terciária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , TemperaturaRESUMO
OBJECTIVES: Irritable bowel syndrome (IBS) is associated with a state of chronic visceral hypersensitivity, but the underlying molecular mechanisms of visceral hyperalgesia remain elusive. This study was designed to examine changes in the excitability and alterations of voltage-gated K+ currents in subpopulations of colonic dorsal root ganglion (DRG) neurons in a rat model of IBS-like visceral hypersensitivity. METHODS: The model of IBS-like visceral hypersensitivity was induced by intracolonic infusion of 0.5% acetic acid (AA) in saline from postnatal days 8 -21. Experiments were conducted when rats became adults. DRG neurons innervating the colon were identified by 1,1'-dioleoyl-3,3,3',3-tetramethylindocarbocyanine methanesulfonate (DiI) fluorescence labeling and were immunostained for isolectin B4 (IB4) binding to classify these colonic neurons. Patch-clamp recordings were made from acutely dissociated DiI-labeled DRG neurons, and the expression of K+ channel in L6-S2 DRG was examined by reverse transcription-polymerase chain reaction (RT-PCR) and western blot. RESULTS: (1) Neonatal AA treatment induced long-lasting visceral hypersensitivity without significant inflammation but with mast cell hyperplasia. (2) Colonic DRG neurons contained IB4-positive and negative neurons with different electrophysiological properties. IB4-positive colonic neurons have longer action potentials (APs) and larger A-type K+ currents (I(A)) than the IB4-negative neurons, and IB4 phenotypic changes of colonic neurons were not involved in the chronic visceral hypersensitivity. (3) Neonatal AA treatment decreased I(A) density and changed the electrophysiological properties of I(A) and I(K) by shifting the steady-state inactivation toward a negative direction in IB4-positive colonic neurons. The excitability of these cells increased. (4) Kv4.3 was downregulated in neonatal AA-treated rats compared with control rats, which suggests a possible mechanism regarding the changes in electrical activity of DRG neurons in these rats. CONCLUSIONS: A new model for chronic visceral hypersensitivity following a diluted AA stimulus in the neonatal period is described. The hypersensitivity may be associated with mast cell hyperplasia in the colon and increased excitability of IB4-positive colonic neurons as a result of suppression of I(A) density and a shift in the inactivation curves of I(A) and I(K) in a hyperpolarizing direction in these cells. This study identifies for the first time a specific molecular mechanism in subpopulations of colonic DRG neurons that underlies chronic visceral hypersensitivity.
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Colo/inervação , Síndrome do Intestino Irritável/fisiopatologia , Canais de Potássio de Abertura Dependente da Tensão da Membrana/fisiologia , Células Receptoras Sensoriais/fisiologia , Vísceras/inervação , Animais , Modelos Animais de Doenças , Glicoproteínas/análise , Lectinas/análise , Ratos , Células Receptoras Sensoriais/química , VersicanasRESUMO
Accumulating evidence indicates that the microbes colonizing human bodies have crucial effects on human health and the discovery of disease-related microbes will promote the discovery of biomarkers and drugs for the prevention, diagnosis, treatment, and prognosis of diseases. However clinical experiments of disease-microbe associations are time-consuming, laborious and expensive, and there are few methods for predicting potential microbe-disease association. Therefore, developing effective computational models utilizing the accumulated public data of clinically validated microbe-disease associations to identify novel disease-microbe associations is of practical importance. We propose a novel method based on the KATZ model and Bipartite Network Recommendation Algorithm (KATZBNRA) to discover potential associations between microbes and diseases. We calculate the Gaussian interaction profile kernel similarity of diseases and microbes based on validated disease-microbe associations. Then, we construct a bipartite graph and execute a bipartite network recommendation algorithm. Finally, we integrate the disease similarity, microbe similarity and bipartite network recommendation score to obtain the final score, which is used to infer whether there are some novel disease-microbe interactions. To evaluate the predictive power of KATZBNRA, we tested it with the walk length 2 using global leave-one-out cross validation (LOOV), two-fold and five-fold cross validations, with AUCs of 0.9098, 0.8463 and 0.8969, respectively. The test results also show that KATZBNRA is more accurate than two recent similar methods KATZHMDA and BNPMDA.
RESUMO
Nogo-66 receptor (NgR), a common receptor for the three known myelin-associated inhibitors, i.e., Nogo-A, myelin-associated glycoprotein (MAG), and oligodendrocyte myelin glycoprotein (OMgp), plays a key role in the failure of axonal regeneration in the adult mammalian central nervous system (CNS). Here we report a novel vaccine approach that stimulates the production of anti-NgR antibody to overcome NgR-mediated growth inhibition after spinal cord injury (SCI). We showed that adult rats immunized with recombinant NgR produced high titers of the anti-NgR antibody and that antisera obtained from the immunized rats promoted neurite outgrowth of rat cerebellar neurons on the inhibitory MAG substrate in vitro. In a spinal cord dorsal hemisection model, NgR immunization promoted regeneration of lesioned corticospinal tract (CST) axons, anterogradely labeled with biotin dextran amine (BDA), beyond the lesion site. In a contusive SCI model, NgR immunization markedly reduced the total lesion volume and improved Basso, Beattie, and Bresnahan (BBB) locomotor rating scale and grid walking performance. Thus, the NgR vaccine approach may represent a promising repair strategy to promote structural and functional recovery following SCI.
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Axônios/fisiologia , Proteínas da Mielina/imunologia , Regeneração Nervosa , Receptores de Superfície Celular/imunologia , Traumatismos da Medula Espinal/terapia , Vacinas Sintéticas/uso terapêutico , Animais , Anticorpos/análise , Anticorpos/sangue , Western Blotting , Células Cultivadas , Cerebelo/citologia , Ensaio de Imunoadsorção Enzimática , Feminino , Proteínas Ligadas por GPI , Humanos , Neuritos/fisiologia , Receptor Nogo 1 , Ratos , Ratos Sprague-Dawley , Recuperação de Função Fisiológica , Medula Espinal/imunologia , Medula Espinal/patologia , Traumatismos da Medula Espinal/patologia , Traumatismos da Medula Espinal/fisiopatologia , VacinaçãoRESUMO
OBJECTIVE: To evaluate the diagnostic efficacy of IFIR-FIESTA technique in detecting renal artery stenosis in elderly patients. METHODS: Twenty-seven aged patients underwent both IFIR-FIESTA and 3D CE-MRA examinations. The imaging quality and renal artery stenosis grades were evaluated. Kappa test was used to assess the consistency between the two methods. With CE-MRA as the reference, the diagnostic sensitivity, specificity, accuracy, PPV and NPV for IFIR-FIESTA were calculated in detecting renal artery stenosis. RESULTS: The images by the two methods were 100% qualified for diagnosis, although the image quality of CE-MRA was significantly better. IFIR-FIESTA and CE-MRA showed excellent consistency in detecting renal artery stenosis. With CE-MRA as the reference, the diagnostic sensitivity, specificity, accuracy, PPV and NPV for IFIR-FIESTA were 97.1%, 100%, 98.1%, 100%, and 95% in detecting renal artery stenosis, respectively. CONCLUSION: IFIR-FIESTA is feasible as a routine examination for detecting renal artery stenosis in elderly patients.
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Meios de Contraste , Angiografia por Ressonância Magnética/métodos , Obstrução da Artéria Renal/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Estudos de Viabilidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Sensibilidade e EspecificidadeRESUMO
Modulation of activation kinetics by divalent ions is one of the characteristic features of Eag channels. Here, we report that Mg(2+)-dependent deceleration of Eag channel activation is significantly attenuated by a G297E mutation, which exhibits a gain-of-function phenotype in Drosophila by suppressing the effect of shaker mutation on behavior and neuronal excitability. The G297 residue is located in the intracellular linker of transmembrane segments S2 and S3, and is thus not involved in direct binding of Mg(2+) ions. Moreover, mutation of the only positively charged residue in the other intracellular linker between S4 and S5 also results in a dramatic reduction of Mg(2+)-dependent modulation of Eag activation kinetics. Collectively, the two mutations in eag eliminate or even paradoxically reverse the effect of Mg(2+) on channel activation and inactivation kinetics. Together, these results suggest an important role of the intracellular linker regions in gating processes of Eag channels.