RESUMEN
In natural populations of Drosophila melanogaster, an amylase isozyme with the lowest alpha-amylase activity (AMY(1,1)) is predominant. To evaluate the selective significance of AMY(1,1) and its regulatory factor(s), we examined selection experiments in laboratory populations on two distinct food environments. After 300 generations, AMY(1,1) became predominant (89%) in a glucose (a product of AMY)-rich environment, while an isozyme with higher alpha-amylase activity, AMY(1,6), became predominant (83%) in a starch (substrate)-rich environment. We found that the identical alleles of the amylase (Amy) gene, which encodes each of AMY(1,1) and AMY(1,6), were shared between the two populations in the different food environments, employing the nucleotide sequencing of the duplicated Amy genes. Nevertheless, AMY(1,6) homozygotes selected in the starch-rich environment had a twofold higher AMY enzyme activity than those selected in the glucose-rich environment, suggesting a coadaptation of the coding region and its regulatory factor(s) on the genetic background. Such a difference in AMY enzyme activity was not detected between AMY(1,1) homozygotes, suggesting that the effect of the genetic background is epistatic. Our results indicate that natural selection is working on the Amy gene system as a whole for flies to adapt to the various food environments of local populations.
Asunto(s)
Drosophila melanogaster/genética , alfa-Amilasas/genética , Adaptación Fisiológica/genética , Animales , Secuencia de Bases , Cartilla de ADN , Proteínas de Drosophila/genética , Drosophila melanogaster/enzimología , Evolución Molecular , Regulación Enzimológica de la Expresión Génica , Isoenzimas/genética , Selección GenéticaRESUMEN
Reverse transcriptase-polymerase chain reaction (RT-PCR) amplification of the nucleocapsid (N) gene of mouse hepatitis virus (MHV) successfully detected 36 strains from the faeces of mice that had been housed in animal facilities in Japan from 2000 to 2003. Subsequent restriction fragment length polymorphism (RFLP) analysis, using the enzymes Acc I, Alu I, EcoR I and Mbo I, demonstrated that these strains could be divided into five distinct subgroups and that the same MHV strains were detected from closely related mouse facilities. Furthermore, strains from the same facility showed the same RFLP pattern, irrespective of the year of detection, but this pattern varied between different locations. This study shows that RFLP analyses are a rapid and useful method for differentiation of MHV strains.
Asunto(s)
Genotipo , Virus de la Hepatitis Murina/genética , Polimorfismo de Longitud del Fragmento de Restricción , Animales , Heces/virología , Ratones , Virus de la Hepatitis Murina/clasificación , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
A series of erythromycin A 9-O-substituted oxime ether derivatives have been synthesized and evaluated for antibacterial activity against Mycobacterium avium complex (MAC) and Staphylococcus aureus. These compounds possessed stronger in vitro activity against MAC including macrolide-resistant strains than clarithromycin (2), although in vitro antibacterial activities of these compounds were less than that of 2 against Staphylococcus aureus. Our studies found that several factors contribute to the antibacterial activity against MAC. The length and spatial orientation of the substituent at 9-position were found to significantly influenced the anti-MAC activity, especially against macrolide-resistant strains. Of all the compounds prepared, erythromycin A 9-[O-(4-phenylbutyl)oxime] (12q) and erythromycin A 9-[O-(3-phenoxypropyl)oxime] (12t) possessed 16 times stronger antibacterial activity than 2 against clarithromycin-resistant strains. Surprisingly, the minimum inhibitory concentrations (MICs) of 12q and 12t against the resistant strains were almost same as those against the susceptible strains. These results suggest that the erythromycin A 9-O-substituted oxime ether derivatives would be promising macrolide antibiotics.
Asunto(s)
Antibacterianos/síntesis química , Antibacterianos/farmacología , Eritromicina/síntesis química , Eritromicina/farmacología , Complejo Mycobacterium avium/efectos de los fármacos , Fenómenos Químicos , Química Física , Fragmentación del ADN , Eritromicina/análogos & derivados , Espectroscopía de Resonancia Magnética , Pruebas de Sensibilidad Microbiana , Modelos Moleculares , Oximas/síntesis química , Oximas/farmacología , Relación Estructura-Actividad Cuantitativa , ARN Bacteriano/efectos de los fármacos , ARN Ribosómico/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacosRESUMEN
The in vivo activity of olamufloxacin (HSR-903), a new fluoroquinolone, was evaluated and compared with ciprofloxacin, sparfloxacin and levofloxacin. Olamufloxacin was active against systemic infection in mice inoculated with both Gram-positive and -negative bacteria. Olamufloxacin had equal efficacy for experimental urinary tract infections in mice caused by Pseudomonas aeruginosa.
Asunto(s)
Antiinfecciosos/uso terapéutico , Infecciones Bacterianas/tratamiento farmacológico , Fluoroquinolonas , Quinolonas/uso terapéutico , Infecciones Urinarias/tratamiento farmacológico , Animales , Masculino , Ratones , Ratones Endogámicos ICRRESUMEN
Samples of feces and vomit collected from patients during 13 non-bacterial gastroenteritis outbreaks which occurred in Hokkaido between 1995 and 1998 were examined by electron microscopy (EM) and reverse-transcription polymerase chain reaction (RT-PCR) for evidence of infection with human caliciviruses (HuCVs). In 6 food-borne outbreaks, oysters were the probable source of infection, while the origin of HuCVs was not found out for the other 7 outbreaks. One-hundred-eleven of 214 stool, vomit and oyster specimens examined gave positive results by RT-PCR, while HuCVs were detected by EM in 36 of 121 stool specimens examined. We determined the nucleotide sequences of 470-bp or 373-bp PCR products amplified from the RNA polymerase region of the HuCV genomes with primer sets MR3/4 and Yuri22F/R, respectively. The sequences of different strains revealed great heterogenicity, with a range of 60 to 100% homology among strains. In a few cases, a mixed genotype was found in the same patient or same outbreak by means of nested PCR and cloning of PCR products into an appropriate vector. Of the 19 different strains found, 4 strains could be classified as Norwalk virus (genogroup 1) and the other 15 strains as Snow Mountain agent (genogroup 2) based on genotyping with homology analysis. Furthermore, the strains belonging to genogroup 2 could be classified into 4 subgroups with more than 93% homology in amino acids among strains in the subgroup.
Asunto(s)
Infecciones por Caliciviridae/virología , Caliciviridae/genética , Caliciviridae/aislamiento & purificación , Gastroenteritis/virología , Animales , Infecciones por Caliciviridae/epidemiología , Cartilla de ADN , ADN Viral/análisis , ADN Viral/genética , Brotes de Enfermedades , Heces/virología , Gastroenteritis/epidemiología , Humanos , Japón/epidemiología , Microscopía Electrónica , Datos de Secuencia Molecular , Ostreidae/virología , Filogenia , Reacción en Cadena de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADNRESUMEN
Following the characterization of the glycosidic constituents in a medical foodstuff "moroheiya," the leaves of Corchorus olitorius L., four higher fatty acids with a trienone function, corchorifatty acids, A, B, C, and D, an undecanoic acid, corchorifatty acid E, and a trihydroxyfatty acid, corchorifatty acid F, were isolated from the less polar fraction of "moroheiya". The structures and optical purity of corchorifatty acids were determined on the basis of chemical and physicochemical evidence. Corchorifatty acids A, B, and C showed an inhibitory effect on lipopolysaccharide-induced NO production in cultured mouse peritoneal macrophages.
Asunto(s)
Inhibidores Enzimáticos/aislamiento & purificación , Ácidos Grasos/aislamiento & purificación , Macrófagos Peritoneales/efectos de los fármacos , Óxido Nítrico Sintasa/antagonistas & inhibidores , Plantas Comestibles/química , Plantas Medicinales/química , Animales , Células Cultivadas , Inhibidores Enzimáticos/farmacología , Ácidos Grasos/farmacología , Japón , Lipopolisacáridos , Macrófagos Peritoneales/enzimología , Ratones , Óxido Nítrico/análisis , Nitritos/análisis , Hojas de la Planta/química , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
The methanol extract of wax gourd (Japanese name "Tougan"), the fruits of Benincasa hispida COGN. (Cucurbitaceae), was found to show inhibitory activity on the histamine release from rat exudate cells induced by antigen-antibody reaction. Through bioassay-guided separation, four known triterpenes and two known sterols were isolated as active components together with a flavonoid C-glycoside, an acylated glucose, and a benzyl glycoside. Among the active triterpenes and sterols, two triterpenes, alnusenol and multiflorenol, were found to potently inhibit the histamine release.
Asunto(s)
Cucurbitaceae/química , Liberación de Histamina/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Células Cultivadas , Depresión Química , Masculino , Mastocitos/metabolismo , Metanol , Cavidad Peritoneal/citología , Extractos Vegetales/aislamiento & purificación , Ratas , Ratas Wistar , Esteroles/aislamiento & purificación , Esteroles/farmacología , Triterpenos/aislamiento & purificación , Triterpenos/farmacologíaRESUMEN
The in vivo activity of HSR-903, a new fluoroquinolone, against major bacteria which cause respiratory tract infections was evaluated. HSR-903 was active against experimental respiratory tract infections in mice challenged with penicillin-susceptible and penicillin-resistant Streptococcus pneumoniae and Haemophilus influenzae strains. Treatment with HSR-903 reduced the bacterial numbers in infected murine lungs. In accord with the pulmonary clearance results, the rates of survival for mice treated with HSR-903, sparfloxacin, levofloxacin, ciprofloxacin, and benzylpenicillin were 50, 30, 10, 0, and 0%, respectively, 14 days after being infected with penicillin-resistant S. pneumoniae. A pharmacokinetic study with pneumonic mice showed that the levels of HSR-903 in the lungs were seven to eight times higher than those in the plasma. These results indicate that clinical studies of HSR-903 against respiratory tract infections may be warranted.
Asunto(s)
Antiinfecciosos/uso terapéutico , Fluoroquinolonas , Infecciones por Haemophilus/tratamiento farmacológico , Haemophilus influenzae , Quinolonas/uso terapéutico , Infecciones del Sistema Respiratorio/tratamiento farmacológico , Infecciones Estreptocócicas/tratamiento farmacológico , Streptococcus pneumoniae , Animales , Antiinfecciosos/farmacocinética , Área Bajo la Curva , Infecciones por Haemophilus/microbiología , Levofloxacino , Ratones , Ratones Endogámicos ICR , Pruebas de Sensibilidad Microbiana , Ofloxacino/farmacocinética , Ofloxacino/uso terapéutico , Quinolonas/farmacocinética , Infecciones del Sistema Respiratorio/microbiología , Infecciones Estreptocócicas/microbiologíaRESUMEN
The methanolic extract of a food garnish "Tonburi", the fruit of Japanese kochia scoparia (L.) Schrad. (Chenopodiaceae), was found to inhibit the increase in serum glucose-loaded rats. Through bioassay-guided separation, momordin Ic and its 2'-O-beta-D-glucopyranoside were isolated as the active principles from this medicinal foodstuff together with three new saponins named scoparianosides A, B, and C. The structures of scoparianosides A, B, and C were elucidated on the basis of chemical and physicochemical evidence as 3 beta, 22 alpha-dihydroxyolean-12en-28-oic acid (22 alpha-hydroxyoleanolic acid), 3-O-beta-D-xylopyranosyl (1 --> 3)-beta-D-glucopyranosiduronic acid, 3 beta-hydroxyolean -18-en-28-oic acid (morolic acid), 3-O-beta-D-xylopyranosyl(1 --> 3)-beta-D -glucopyranosiduronic acid, and 3 beta-hydroxyolean-13(18)-en-28-oic acid, 3-O-beta-D-xylopyranoxyl(1 --> 3) -beta-D-glucopyranosiduronic acid. Momordin Ic and its 2'-O-beta-D-glucopyranoside, both of which are the principal saponin constituents of this medicinal foodstuffs, were found to potently inhibit glucose and ethanol absorption in rats.
Asunto(s)
Depresores del Sistema Nervioso Central/farmacocinética , Etanol/farmacocinética , Frutas/química , Glucosa/farmacocinética , Hipoglucemiantes/aislamiento & purificación , Ácido Oleanólico/análogos & derivados , Plantas Medicinales/química , Saponinas/aislamiento & purificación , Saponinas/farmacología , Animales , Depresores del Sistema Nervioso Central/sangre , Fenómenos Químicos , Química Física , Chenopodiaceae/química , Etanol/sangre , Hidrólisis , Hipoglucemiantes/farmacología , Japón , Espectroscopía de Resonancia Magnética , Masculino , Ratas , Ratas Wistar , Relación Estructura-ActividadRESUMEN
Three new ionone glucosides named corchoionosides A, B, and C were isolated from the leaves of Corchorus olitorius, commonly called "moroheiya" in Japanese, together with seven known compounds, an ionone glucoside (6S,9R)-roseoside, a monoterpene glucoside betulalbuside A, two flavonol glucosides astragalin and isoquercitrin, two coumarin glucosides scopolin and cichoriine, and chlorogenic acid. The absolute stereostructures of corchoionosides A, B, and C were determined by chemical and physiochemical evidence, which included the result of application of a modified Mosher's method, the CD helicity rule, and chemical correlation with (6S,9R)-roseoside. Corchoionosides A and B and (6S,9R)-roseoside were found to inhibit the histamine release from rat peritoneal exudate cells induced by antigen-antibody reaction.
Asunto(s)
Glucósidos/química , Liberación de Histamina , Extractos Vegetales/química , Animales , Reacciones Antígeno-Anticuerpo , Glucósidos/farmacología , Liberación de Histamina/efectos de los fármacos , Modelos Moleculares , Ratas , EstereoisomerismoRESUMEN
The methanol-soluble fraction from a Chinese natural medicine Hoveniae Semen Seu Fructus, the seed and fruit of Hovenia dulcis THUNB. (Rhamnaceae) was found to show an inhibitory effect on the alcohol-induced muscular relaxation and a protective activity on the D-galactosamine/lipopolysaccharide or carbon tetrachloride-induced liver injury. Through bioassay-guided separation using a traction performance test, three new dihydrofravonols named hovenitins I, II, and III were isolated from Hoveniae Semen Seu Fructus together with four known flavonoids, (+)-ampelopsin, laricetrin, myricetin, and (+)-gallocatechin. The absolute stereostructures of hovenitins I, II, and III were determined on the basis of chemical and physicochemical evidence to be (2R, 3R)-5,7,4',5'-tetrahydroxy-3'-methoxydihydroflavonol, (2R,3S)-5,7,4',5'-tetrahydroxy-3'-methoxy-dihydroflavonol, and (2R, 3S)-5,7,3',4',5'-pentahydroxydihydro-flavonol, respectively. Hovenitin I and (+)-ampelopsin, both of which were principal ingredients of the active fractions from this natural medicine, were found to show an inhibitory activity on the ethanol-induced muscle relaxation in rats. In addition, hovenitin I showed a protective activity on the liver injury induced by D-galactosamine/lipopolysaccharide or carbon tetrachloride in mice.
Asunto(s)
Enfermedad Hepática Inducida por Sustancias y Drogas/tratamiento farmacológico , Medicamentos Herbarios Chinos/farmacología , Etanol/antagonistas & inhibidores , Flavonoides/farmacología , Flavonoles , Relajación Muscular/efectos de los fármacos , Músculo Esquelético/efectos de los fármacos , Animales , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/uso terapéutico , Etanol/farmacocinética , Flavonoides/química , Flavonoides/uso terapéutico , Absorción Intestinal/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos , Ratas , Ratas Wistar , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Acylated polyhydroxyolean-12-ene triterpene oligoglycosides, camelliasaponins A1, A2, B1, B2, C1, and C2, were isolated from the seeds of Camellia japonica L. The structures of six camelliasaponins were elucidated on the basis of chemical and physicochemical evidence. Camelliasaponins B1, B2, C1, and C2 were found to exhibit inhibitory activity on ethanol absorption. By comparison of the inhibitory activities for camelliasaponins with those for desacyl-camelliasaponins, acyl groups such as the angeloyl or tigloyl group were found to be essential to exerting the activity.
Asunto(s)
Depresores del Sistema Nervioso Central/farmacocinética , Etanol/farmacocinética , Saponinas/química , Té/química , Animales , Secuencia de Carbohidratos , Fenómenos Químicos , Química Física , Depresión Química , Absorción Intestinal/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Masculino , Datos de Secuencia Molecular , Ratas , Ratas Wistar , Saponinas/farmacología , Semillas/química , Relación Estructura-ActividadRESUMEN
From the medicinal foodstuff "kuwai", the tuber of Sagittaria trifolia L., four bioactive diterpene ketones, trifoliones A, B, C, and D, two diterpene glucosides, sagittariosides a and b, and a nitroethylphenol glycoside, arabinothalictoside, were isolated, together with six known diterpenes. Their absolute stereostructures were determined on the basis of chemical and physicochemical evidence which included the application of a modified Mosher's method and an exciton chirality method. Among the diterpene constituents, trifoliones A, B, C, and D exhibited inhibitory effects on the histamine release from rat mast cells induced by compound 48/80 and calcium ionophore A-23187.
Asunto(s)
Diterpenos/aislamiento & purificación , Glucósidos/aislamiento & purificación , Glicósidos/aislamiento & purificación , Antagonistas de los Receptores Histamínicos/aislamiento & purificación , Raíces de Plantas/química , Plantas Comestibles/química , Plantas Medicinales/química , Animales , Calcimicina/antagonistas & inhibidores , Calcimicina/farmacología , Cromatografía Líquida de Alta Presión , Diterpenos/química , Diterpenos/farmacología , Glucósidos/química , Glucósidos/farmacología , Glicósidos/química , Glicósidos/farmacología , Antagonistas de los Receptores Histamínicos/química , Antagonistas de los Receptores Histamínicos/farmacología , Liberación de Histamina/efectos de los fármacos , Técnicas In Vitro , Ionóforos/farmacología , Cetonas/química , Cetonas/aislamiento & purificación , Cetonas/farmacología , Masculino , Mastocitos/efectos de los fármacos , Mastocitos/metabolismo , Medicina Tradicional China , Ratas , Ratas Wistar , Espectrometría de Masa Bombardeada por Átomos Veloces , Espectrofotometría Infrarroja , Estereoisomerismo , p-Metoxi-N-metilfenetilamina/farmacologíaRESUMEN
To understand the mode of persistent infection of Seoul virus in rodents, we examined the distribution of the virus genome and antibody production in infected rats. When 1-day-old rats were inoculated with the KI-83-262 strain, the S segment of viral genome was detected in sera, clots, lungs and kidneys from 3 to 184 days post inoculation (d.p.i.) by nested reverse transcriptase PCR. On the other hand, when 7-week-old rats were infected with this virus, viral genome was detected only in the lungs from 3 to 50 d.p.i. The neutralizing antibody titers of rats inoculated at 1-day of age were higher than those of rats inoculated at 7 weeks of age. In both age groups, however, the IgG avidity of antibody increased along with the course of infection. We found that urban rats (Rattus norvegicus) infected early in life harbored the virus for more than 6 months.
Asunto(s)
Infecciones por Hantavirus/inmunología , Infecciones por Hantavirus/transmisión , Orthohantavirus/inmunología , Orthohantavirus/aislamiento & purificación , Animales , Anticuerpos Antivirales/análisis , Anticuerpos Antivirales/inmunología , Afinidad de Anticuerpos , Células Cultivadas , Chlorocebus aethiops , Reservorios de Enfermedades , Genes Virales , Genoma Viral , Orthohantavirus/genética , Infecciones por Hantavirus/genética , Inmunoglobulina G/inmunología , Riñón/virología , Pulmón/virología , Pruebas de Neutralización , Reacción en Cadena de la Polimerasa , ARN Viral/aislamiento & purificación , Ratas , Ratas Wistar , Sensibilidad y Especificidad , Factores de Tiempo , Células Vero , Proteínas Virales/inmunologíaRESUMEN
Five new saponins named elatosides G, H, I, J, and K were isolated from a garnish foodstuff "Taranome," the young shoot of Aralia elata SEEM., together with hederagenin-3-O-glucuronopyranoside and elatoside C. Their chemical structures were elucidated on the basis of chemical and physicochemical evidence. Elatosides G, H, and I were found to exhibit potent hypoglycemic activity in the oral glucose tolerance test in rats.
Asunto(s)
Hipoglucemiantes/aislamiento & purificación , Ácido Oleanólico/análogos & derivados , Saponinas/aislamiento & purificación , Análisis de Varianza , Animales , Fraccionamiento Químico , Cromatografía Líquida de Alta Presión , Prueba de Tolerancia a la Glucosa , Hipoglucemiantes/química , Hipoglucemiantes/toxicidad , Espectroscopía de Resonancia Magnética , Masculino , Ratas , Ratas Wistar , Saponinas/química , Espectrometría de Masa Bombardeada por Átomos Veloces , Relación Estructura-ActividadRESUMEN
We conducted field surveys of indigenous rodent species in Hokkaido, Japan from 1980 to 1993. Serum samples were collected from 663 rodents, including Clethrionomys rufocanus, Apodemus speciosus, A. argenteus, and C. rutilus. Antibody to hantavirus was determined by the protein G antibody assay. Positive C. rufocanus were detected in seven of eight collection sites, but no antibody was detected in the remaining rodent species. To reveal the serotype of the circulating virus in C. rufocanus, antibody titers to Hantaan, Seoul, Puumala, and Prospect Hill viruses were compared by means of the focus reduction neutralization test. The titers in positive sera were extremely high to the Sotkamo strain of Puumala virus. Results were confirmed by the reverse transcriptase-polymerase chain reaction, and suggested that Puumala-related viruses are in circulation among C. rufocanus populations in Hokkaido.