RESUMO
The following two matters should be resolved in order for biosimulation tools to be accepted by users in biology/medicine: (1) remove issues which are irrelevant to biological importance, and (2) allow users to represent biopathways intuitively and understand/manage easily the details of representation and simulation mechanism. From these criteria, we firstly define a novel notion of Petri net called Hybrid Functional Petri Net (HFPN). Then, we introduce a software tool, Genomic Object Net, for representing and simulating biopathways, which we have developed by employing the architecture of HFPN. In order to show the usefulness of Genomic Object Net for representing and simulating biopathways, we show two HFPN representations of gene regulation mechanisms of Drosophila melanogaster (fruit fly) circadian rhythm and apoptosis induced by Fas ligand. The simulation results of these biopathways are also correlated with biological observations. The software is available to academic users from http://www.GenomicObject.Net/.
Assuntos
Simulação por Computador , Drosophila melanogaster , Animais , Regulação da Expressão Gênica , Genômica , Modelos Biológicos , Transdução de Sinais , SoftwareRESUMO
We investigated the effect of substances present in Japanese sake on the response of ionotropic γ-aminobutyric acid (GABA)A receptors expressed in Xenopus oocytes. Sake was fractionated by ion-exchange chromatography. The fraction containing organic acids (OA fraction) showed agonist activities on the GABAA receptor. OA fractions from sake were analyzed by capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS). Of the 64 compounds identified, 13 compounds showed GABAA receptor agonist activities. Especially, l-lactic acid showed high agonist activity and its EC50 value was 37µM. Intraperitoneal injections of l-lactic acid, gluconic acid, and pyruvic acid (10, 10, and 5mg/kg BW, respectively), which showed agonistic activity on the GABAA receptor, led to significant anxiolytic effects during an elevated plus-maze test in mice.
Assuntos
Bebidas Alcoólicas/análise , Agonistas de Receptores de GABA-A/farmacologia , Ácido Láctico/farmacologia , Oócitos/efeitos dos fármacos , Receptores de GABA-A/metabolismo , Animais , Cromatografia por Troca Iônica , Feminino , Oócitos/metabolismo , Xenopus laevisRESUMO
Beer induced the response of the ionotropic gamma-aminobutyric acid receptors (GABA(A) receptors) expressed in Xenopus oocytes, indicating the presence of gamma-aminobutyric acid (GABA)-like activity. Furthermore, the pentane extract of the beer, hop (Humulus lupulus L.) oil, and myrcenol potentiated the GABA(A) receptor response elicited by GABA. The GABA(A) receptor responses were also potentiated by the addition of aliphatic esters, most of which are reported to be present in beer flavor. Aliphatic esters showed the tendency to decrease in the potentiation of the GABA(A) receptor response with an increase in their carbon chain length. When myrcenol was injected to mice prior to intraperitoneal administration of pentobarbital, the pentobarbital-induced sleeping time of mice increased additionally. Therefore, the beer contained not only GABA-like activity but also the modulator(s) of the GABA(A) receptor response.
Assuntos
Cerveja/análise , Humulus/química , Receptores de GABA-A/efeitos dos fármacos , Monoterpenos Acíclicos , Alcenos/farmacologia , Animais , Sinergismo Farmacológico , Ésteres/farmacologia , Feminino , Expressão Gênica , Camundongos , Monoterpenos/farmacologia , Oócitos/metabolismo , Óleos de Plantas/farmacologia , Receptores de GABA-A/genética , Transfecção , Xenopus laevis , Ácido gama-Aminobutírico/farmacologiaRESUMO
A cDNA, which has a high homology with teleost Platichthys flesus [Arg8] vasotocin (AVT) receptor (GenBank: AK033957), was found in mouse genome database. Analyses of the deduced amino acid sequence revealed that a cDNA has several features of AVT receptor. We tentatively named it as a mouse vasotocin receptor (MVTR). A two-electrodes voltage clamp technique was applied to characterize the MVTR expressed in Xenopus laevis oocytes. AVT induced Ca2+-dependent Cl- currents in Xenopus oocytes injected with MVTR cRNA. On the other hand, [Arg8] vasopressin, oxytocin and isotocin did not induce such currents. RT-PCR showed that MVTR mRNA was specifically expressed in the brain. In situ hybridization analysis demonstrated significant expression of MVTR mRNA in suprachiasmatic nucleus, arcuate nucleus and medial habenular nucleus of mouse brain. These results suggest that MVTR may mediate a variety of physiological functions in mouse.
Assuntos
Receptores de Vasopressinas/química , Receptores de Vasopressinas/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Regulação da Expressão Gênica , Camundongos , Dados de Sequência Molecular , Oócitos/metabolismo , Receptores de Vasopressinas/genética , Xenopus laevisRESUMO
OBJECTIVE: The purpose of this study was to determine whether anterior crowns fabricated using a computerized shade selection system (ShadeScan, Cynovad) (experimental procedure) match adjacent teeth better than anterior crowns fabricated using conventional shade prescription and clinical slides (control). METHOD AND MATERIALS: Five subjects who required a crown to restore a maxillary central incisor were selected. Two metal-ceramic crowns were fabricated for each incisor, 1 using the experimental procedure and 1 using the control method. The shade selection method to be used for the first and second crowns was randomly assigned. The duration of each procedure was recorded. Each restoration was tried-in in a double-blind manner and evaluated for its level of match to adjacent teeth using modified Ryge criteria. Data were analyzed within each subject using descriptive statistics and paired t test (alpha = .05). RESULTS: In 40% of the cases, both procedures did equally well. In the remaining 60% of the cases the control procedure (two-thirds of the cases) performed better than the experimental procedure (one-third of the cases). Duration of the control procedure was 14.4 +/- 5 minutes, and the experimental procedure was 5.2 +/- 3.3 minutes. A paired t test showed the difference was significant (P = .0045). CONCLUSION: The level of matching of crowns fabricated with the ShadeScan system was not different from crowns fabricated using the control. However, it took significantly less time to record the shade with the ShadeScan system.
Assuntos
Coroas , Tomada de Decisões Assistida por Computador , Pigmentação em Prótese , Cor , Estudos Cross-Over , Método Duplo-Cego , Humanos , Ligas Metalo-Cerâmicas/química , Projetos PilotoRESUMO
Recently, 3-O-octanoyl-(+)-catechin (OC) was synthesized from (+)-catechin (C) by incorporation of an octanoyl chain into C in the light of (-)-epicatechin gallate (ECg) and (-)-epigallocatechin gallate (EGCg), which are the major polyphenols found in green tea and have strong physiological activities. OC was found to inhibit the response of ionotropic gamma-aminobutyric acid (GABA) receptors (GABA(A) receptors) and Na+/glucose cotransporters expressed in Xenopus oocytes in a noncompetitive manner more strongly than does C. OC also induced a nonspecific membrane current and decreased the membrane potential of the oocyte, and thus death of the oocyte occurred even at lower concentrations than that induced by C or EGCg. Although EGCg produced H2O2 in aqueous solution, OC did not. This newly synthesized catechin derivative OC possibly binds to the lipid membrane more strongly than does C, Ecg, or EGCg and as a result perturbs the membrane structure.
Assuntos
Catequina/análogos & derivados , Catequina/farmacologia , Potenciais da Membrana/efeitos dos fármacos , Proteínas de Transporte de Monossacarídeos/efeitos dos fármacos , Oócitos/metabolismo , Oócitos/ultraestrutura , Receptores de GABA-A/efeitos dos fármacos , Animais , Bovinos , Expressão Gênica , Peróxido de Hidrogênio/metabolismo , Proteínas de Transporte de Monossacarídeos/genética , Proteínas de Transporte de Monossacarídeos/fisiologia , Receptores de GABA-A/genética , Receptores de GABA-A/fisiologia , Proteínas Recombinantes , Xenopus laevisRESUMO
1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of Japanese whiskey after various aging periods in oak barrels was measured to evaluate the antioxidative effects of whiskey. The activity of the whiskey increased with the aging period with high correlation. The activity of various types of whiskey was measured and shown to be correlated to the potentiation of the GABAA receptor response measured in a previous paper. However, the fragrant compounds in the whiskey which potentiated the GABAA receptor response had low DPPH radical scavenging activity, while phenol derivatives had high radical scavenging activity. The whiskey was extracted by pentane. The aqueous part showed the scavenging activity, whereas the pentane part did not. Thus, both the DPPH radical scavenging activity and the potentiation of the GABAA receptor response increased during whiskey aging in oak barrels, but were due to different components. The whiskey protected the H2O2-induced death of E. coli more than ethanol at the same concentration as that of the whiskey. The changes that occurred in the whiskey during aging may be the reason aged whiskies are so highly valued.
Assuntos
Bebidas Alcoólicas/análise , Sequestradores de Radicais Livres/análise , Picratos/química , Compostos de Bifenilo , Sequestradores de Radicais Livres/química , Quercus , Fatores de Tempo , MadeiraRESUMO
The effects of both coffee components and coffee extract on the electrical responses of GABA(A) receptors expressed in Xenopus oocytes were studied by injecting cRNAs of the alpha(1) and beta(1) subunits of the bovine receptors. The aqueous extract of coffee dose-dependently inhibited the GABA-elicited responses, whereas the lipophilic extract of coffee by diethyl ether slightly potentiated it at low doses (0.1-0.4 microL/mL) but showed inhibition at high doses (0.5-0.8 microL/mL). Theophylline inhibited the response in a noncompetitive mechanism (K(i) = 0.55 mM), whereas theobromine and trigonelline hydrochloride inhibited it in a competitive manner, K(i) = 3.8 and 13 mM, respectively. Benzothiazole, catechol, 2,4-dimethylstyrene, guaiacol, 1-octen-3-ol, sotolone, and 2,3,5-trimethylphenol potentiated the responses significantly. Potentiation elicited by guaiacol and sotolone was independent of GABA concentrations, whereas that by 1-octen-3-ol was dependent. When 1-octen-3-ol (100 mg/kg) was orally administered to mice prior to intraperitoneal administration of pentobarbital, the sleeping time of mice induced by pentobarbital increased significantly.
Assuntos
Coffea/química , Extratos Vegetais/farmacologia , Receptores de GABA-A/efeitos dos fármacos , Alcaloides/farmacologia , Animais , Bovinos , Sinergismo Farmacológico , Feminino , Antagonistas GABAérgicos/farmacologia , Expressão Gênica , Oócitos/metabolismo , Receptores de GABA-A/genética , Receptores de GABA-A/fisiologia , Teobromina/farmacologia , Teofilina/farmacologia , Xenopus laevis , Ácido gama-Aminobutírico/farmacologiaRESUMO
It is known that the target of most mood-defining compounds such as ethanol is an ionotropic gamma-aminobutyric acid receptor (GABA(A) receptor). The potentiation of the response of these inhibitory neurotransmitter receptors induces anxiolytic, sedative, and anesthetic activities in the human brain. Because both extracts of whiskey by pentane and fragrant components in whiskey potentiate the GABA(A) receptor-mediated response, GABA(A) receptors were expressed in Xenopus oocyte by injecting cRNAs prepared from the cloned cDNA for the alpha(1) and beta(1) subunits of the bovine receptors in order to study the effects of whiskey itself on the GABA(A) receptor-mediated response. Whiskey itself also potentiated the electrical responses of GABA(A) receptors generally more than ethanol at the same concentration as that of the whiskey. The potentiation of the GABA(A) receptor-mediated response increased with the aging period of the whiskey. Inhalation of whiskey to mice increased the sleeping time induced by pentobarbital more than that of the same concentration of ethanol as the whiskey. These results suggest that not only ethanol but also minor components in whiskey play an important role in the potentiation of GABA(A) receptor-mediated response and possibly the sedative effect of whiskey. Although the minor components are present in extremely small quantities compared with ethanol in alcoholic beverages, they may modulate the mood or consciousness of humans through the potentiation of the GABA(A) receptor response after absorption into the brain, because these hydrophobic compounds are easily absorbed into the brain across the blood-brain barrier and are several thousands times as potent as ethanol in the potentiation of the GABA(A) receptor-mediated response.
Assuntos
Bebidas Alcoólicas/análise , Receptores de GABA-A/fisiologia , Animais , Bovinos , Eletrofisiologia , Etanol/farmacologia , Feminino , Manipulação de Alimentos , Expressão Gênica , Japão , Masculino , Camundongos , Oócitos/metabolismo , Pentobarbital/farmacologia , Receptores de GABA-A/genética , Sono/efeitos dos fármacos , Fatores de Tempo , Transfecção , Madeira , Xenopus laevisRESUMO
To study the effects of tea components on ionotropic gamma-aminobutyric acid (GABA) receptor response, ionotropic GABA receptors (GABA(A) receptors) were expressed in Xenopus oocytes by injecting cRNAs synthesized from cloned cDNAs of the alpha(1) and beta(1) subunits of the bovine receptors, and their electrical responses were measured by a voltage clamping method. Extracts of green tea, black tea, and oolong tea in an aqueous solution induced the GABA-elicited response, which showed that these teas contain GABA, whereas coffee does not. Caffeine weakly inhibited the response in a competitive manner (K(i) = 15 mM), and (+)-catechin inhibited it in a noncompetitive one (K(i) = 1.7 mM). Especially, two catechin derivatives, (-)-epicatechin gallate and (-)-epigallocatechin gallate, inhibited the response strongly. Alcohols such as leaf alcohol or linalool potentiated the response, possibly because their binding to the potentiation site enhances the GABA-binding affinity to GABA(A) receptors when they bind. Extracts of green tea made with ethyl ether, which must contain lipophilic components of green tea, inhibited the response elicited by GABA, possibly because the amounts of caffeine and catechin derivatives were much larger than fragrant alcohols in such extracts of tea.
Assuntos
Monoterpenos , Oócitos/metabolismo , Extratos Vegetais/farmacologia , Receptores de GABA-A/efeitos dos fármacos , Receptores de GABA-A/fisiologia , Chá/química , Xenopus laevis , Monoterpenos Acíclicos , Animais , Catequina/análogos & derivados , Catequina/farmacologia , Sinergismo Farmacológico , Feminino , Expressão Gênica , Potenciais da Membrana/efeitos dos fármacos , RNA Complementar/genética , Proteínas Recombinantes , Terpenos/farmacologia , Transfecção , Ácido gama-Aminobutírico/farmacologiaRESUMO
It is well-known that the target of most mood-defining compounds is an ionotropic gamma-aminobutyric acid receptor (GABA(A) receptor). The potentiation of the response of these inhibitory neurotransmitter receptors induces anxiolytic, sedative, and anesthetic activity in the human brain. To study the effects of whiskey fragrance on the GABA(A) receptor-mediated response, GABA(A) receptors were expressed in Xenopus oocyte by injecting rat whole brain mRNA or cRNA prepared from the cloned cDNA for the alpha(1) and beta(1) subunits of the bovine receptors. Most whiskey components such as phenol, ethoxy, and lactone derivatives potentiated the electrical responses of GABA(A) receptors, especially ethyl phenylpropanoate (EPP), which strongly potentiated the response. When this compound was applied to mice through respiration, the convulsions induced by pentetrazole were delayed, suggesting that EPP was absorbed by the brain, where it could potentiate the GABA(A) receptor responses. The extract of other alcoholic drinks such as wine, sake, brandy, and shochu also potentiated the responses to varying degrees. Although these fragrant components are present in alcoholic drinks at low concentrations (extremely small quantities compared with ethanol), they may also modulate the mood or consciousness of the human through the potentiation of the GABA(A) receptor response after absorption into the brain, because these hydrophobic fragrant compounds are easily absorbed into the brain through the blood-brain barrier and are several thousands times as potent as ethanol in the potentiation of the GABA(A) receptor-mediated response.
Assuntos
Bebidas Alcoólicas , Odorantes , Receptores de GABA-A/fisiologia , Animais , Química Encefálica , Bovinos , Sinergismo Farmacológico , Feminino , Antagonistas GABAérgicos , Expressão Gênica , Lactonas/farmacologia , Masculino , Oócitos/metabolismo , Pentilenotetrazol , Fenóis/farmacologia , Fenilpropionatos/farmacologia , Ratos , Ratos Wistar , Receptores de GABA-A/efeitos dos fármacos , Receptores de GABA-A/genética , Convulsões/induzido quimicamente , Transfecção , Xenopus laevisRESUMO
To study the effects of polyphenols on the Na(+)/glucose cotransporter (SGLT1) response, SGLT1 was expressed in Xenopus oocytes by injecting cRNA synthesized from the cloned cDNA of the small intestine cotransporter of rats, and the electrical response elicited by glucose or galactose was measured by a voltage clamping method. Most phenol derivatives had no effect on the response. However, the polyphenols (+)-catechin, (-)-epicatechin gallate (ECg), and (-)-epigallocatechin gallate (EGCg), which are components of green tea, caused an inhibition of the response, which was almost independent of glucose concentration. The inhibition constants were estimated to be 2.3 mM for (+)-catechin and 0.45 mM for both ECg and EGCg, assuming the noncompetitive inhibition mechanism. Saponin prepared from tea seeds also inhibited the response significantly. Tannic acid and aqueous extracts of teas induced nonspecific electrical responses in both cRNA-injected and noninjected oocytes at lower concentrations than those that caused an inhibition of the SGLT1 response when their dose-dependent effects were examined. These results are possibly helpful in the development of a dietary supplement for diabetic patients.
Assuntos
Flavonoides , Expressão Gênica , Glicoproteínas de Membrana/antagonistas & inibidores , Proteínas de Transporte de Monossacarídeos/antagonistas & inibidores , Oócitos/metabolismo , Fenóis/farmacologia , Polímeros/farmacologia , Xenopus laevis , Animais , Catequina/análogos & derivados , Catequina/farmacologia , Membrana Celular/fisiologia , Condutividade Elétrica , Feminino , Galactose/farmacologia , Glucose/farmacologia , Intestino Delgado/química , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Proteínas de Transporte de Monossacarídeos/genética , Proteínas de Transporte de Monossacarídeos/metabolismo , Oócitos/fisiologia , Oócitos/ultraestrutura , Técnicas de Patch-Clamp , Ratos , Saponinas/farmacologia , Transportador 1 de Glucose-Sódio , Chá/químicaRESUMO
AIM: To study the synthesis of 5-(3'-indolyl)-oxazoles and their antioxidative activity. METHODS: The amides were prepared from tryptophan and different acid derivatives by the catalytic dehydration of dicyclohexyl carbodiimide (DCC). The characteristic heterocyclic ring system of 5-(3'-indolyl)-oxazoles was constructed by oxidative cyclization of amide, using dicholorodicyanoquinone (DDQ). Their antioxidative activity in vitro was tested using DPPH system. RESULTS: Eleven 2-substituted phenyl-5-(3'-indolyl)-oxazoles were prepared, the compounds 21 and 22 have shown antioxidative activity 3-4 times stronger than that of Vit E, and the compound 29 showed antioxidative activity almost as same as Vit E. CONCLUSION: Three 5-(3'-indoyl)-oxazole compounds synthesized showed potent antioxidative effect and they would be a good antioxidants.
Assuntos
Antioxidantes/síntese química , Indóis/isolamento & purificação , Oxazóis/isolamento & purificação , Antioxidantes/química , Indóis/química , Estrutura Molecular , Oxazóis/químicaRESUMO
We electrophysiologically investigated the effect of some fragrant compounds in oolong tea on the response of ionotropic gamma-aminobutyric acid (GABA) receptors (GABAA receptors) which were expressed in Xenopus oocytes. Of the tested fragrances in oolong tea, cis-jasmone, jasmine lactone, linalool oxide and methyl jasmonate significantly potentiated the response. Among these, cis-jasmone and methyl jasmonate potently potentiated the response, having a respective dissociation constant of the compound (Kp) and maximum potentiation (Vm) of 0.49 mM and 322% for cis-jasmone, and 0.84 mM and 450% for methyl jasmonate. Inhalation of 0.1% cis-jasmone or methyl jasmonate significantly increased the sleeping time of mice induced by pentobarbital, suggesting that these fragrant compounds were absorbed by the brain and thereby potentiated the GABAA receptor response. Both of these compounds may therefore have a tranquillizing effect on the brain.
Assuntos
Óleos Voláteis/farmacologia , Receptores de GABA-A/efeitos dos fármacos , Chá/química , Acetatos/farmacologia , Monoterpenos Acíclicos , Animais , Bovinos , Ciclopentanos/farmacologia , Relação Dose-Resposta a Droga , Eletrofisiologia , Camundongos , Monoterpenos/farmacologia , Oócitos , Oxilipinas , Receptores de GABA-A/genética , Sono/efeitos dos fármacos , Transfecção , XenopusRESUMO
The following two matters should be resolved in order for biosimulation tools to be accepted by users in biology/medicine: (1) remove issues which are irrelevant to biological importance, and (2) allow users to represent biopathways intuitively and understand/manage easily the details of representation and simulation mechanism. From these criteria, we firstly define a novel notion of Petri net called Hybrid Functional Petri Net (HFPN). Then, we introduce a software tool, Genomic Object Net, for representing and simulating biopathways, which we have developed by employing the architecture of HFPN. In order to show the usefulness of Genomic Object Net for representing and simulating biopathways, we show two HFPN representations of gene regulation mechanisms of Drosophila melanogaster (fruit fly) circadian rhythm and apoptosis induced by Fas ligand. The simulation results of these biopathways are also correlated with biological observations. The software is available to academic users from http://www.GenomicObject.Net/.