RESUMO
Strains of microorganisms capable of 7alpha-dehydroxylation of chenodeoxycholate were isolated from rat and human feces. All the strains were strictly anaerobic, non-motile, moderately themioresistant Gram-positive rods. They showed some saccharolytic properties with the production of both acid and gas. They were H(2)S-positive but indole-, skatole-, citrate-, catalase-, and oxidase-negative. The isolated strains capable of 7alpha-dehydroxylation of chenodeoxycholate were also able to oxidize the hydroxyl groups at C-3 and C-7 to keto groups. The following metabolites were isolated: 3-keto-7alpha-hydroxy-5beta-cholanoic acid, 3alpha-hydroxy-7-keto-5beta-cholanoic acid, 3alpha-hydroxy-5beta-cholanoic acid, and 3-keto-5beta-cholanoic acid. The isolated strains did not have the enzymes necessary for hydrolyzing conjugated bile acids. In mixed anaerobic cultures of fecal microorganisms, extensive reduction of the 3-keto group to the 3beta-hydroxyl group occurred. The microorganism(s) responsible for this reaction have as yet not been isolated.
Assuntos
Ácidos e Sais Biliares/metabolismo , Fezes , Mucosa Intestinal/metabolismo , Técnicas Bacteriológicas , Isótopos de Carbono , Cromatografia , Técnicas In Vitro , Oxigenases de Função MistaRESUMO
Germfree rats monocontaminated with the anaerobic microorganisms Clostridium difficile or another Clostridium species (strain G 62) produce auto-antibodies to colon antigen. The antigen can be extracted with phenol water from the feces of germfree rats. Antibodies, demonstrable by means of passive hemagglutination of antigen sensitized sheep erythrocytes appear after monocontamination for 35 days or longer. The indirect immunofluorescence techniques, applied to sections of germfree rat colon, gave positive mucosal staining. The staining was similar to that obtained with sera from patients with ulcerative colitis or from rats immunized with rabbit colon. No antibodies were found in the sera of germfree rats, germfree rats monocontaminated with various other bacteria, conventional rats of germfree origin, or conventional Sprague-Dawley rats. Although the anti-colon antibodies of the Clostridium infected rats reacted with the same feces extract as the antibodies of ulcerative colitis patients or of rabbit colon immunized rats, their specificity was different. While the latter cross-react with polysaccharide from E. coli O14, those from the Clostridium-infected exgermfree rats did not. Rats monocontaminated with Cl. difficile also developed antibodies to this organism, but no cross-reaction between Cl. difficile antigen and colon antigen could be demonstrated. This speaks against breakage of tolerance by cross-reacting bacterial antigen as the cause of autoimmunity in these rats. Other possible mechanisms for autoantibody production in this model are immunogenic alteration of gastrointestinal mucins by bacterial degradation, adjuvant effects of bacterial products, or both.
Assuntos
Autoanticorpos , Infecções por Clostridium/imunologia , Colo/imunologia , Vida Livre de Germes , Animais , Antígenos/isolamento & purificação , Escherichia coli/imunologia , Fezes/análise , Imunofluorescência , Polissacarídeos Bacterianos , RatosRESUMO
The incidence and height of antibody titers to colon, assayed by indirect hemagglutination with a heat stable colon extract from germ free rats, is significantly higher in sera from patients with ulcerative colitis than in those from healthy controls or from patients with amebic liver abscess or dysentery. While sera from ulcerative colitis patients and controls are indistinguishable in regard to incidence and height of antibody titers to Forsman antigen, Staphylococcus aureus S 209, Clostridium difficile, and several common strains of E. coli, they have elevated titers and increased incidence of antibodies to a heat stable antigen of E. coli O14. Patients with amebic dysentery have normal titers of such antibodies. Absorption of patients' sera with E. coli O14 antigen inhibits the colon directed hemagglutination reaction in approximately 30% of the cases tested. Likewise, the anti-E. coli O14 reaction can sometimes be inhibited with the colon extract. Other E. coli strains and other bacteria are inactive or have only weak inhibitory activity. Hemagglutination inhibition experiments show that germ free rat colon and E. coli O14 contain common structures, depicted by antibodies in the patients' sera. This pattern of reactivity closely resembles that seen in rats made autoimmune to colon by injection of newborn rabbit colon. E. coli O14 is known to carry a heterogenetic antigen present in lower concentration (or activity) in most Enterobacteriaceae. Hemagglutination inhibition experiments with rabbit antisera to E. coli O14 suggest that the antigen common for E. coli O14 and colon is related to this heterogenetic antigen. The findings imply that this antigen, which is constantly present in low concentrations in the human colon, may give rise to anticolon antibody formation in ulcerative colitis through breakage of tolerance. Since this antigen is present in healthy individuals as well, additional factors are required to explain the induction of anti-colon autoimmunity in ulcerative colitis.
Assuntos
Formação de Anticorpos , Autoanticorpos/biossíntese , Colite Ulcerativa/imunologia , Animais , Reações Antígeno-Anticorpo , Povo Asiático , População Negra , Clostridium/imunologia , Colo/imunologia , Disenteria Amebiana/imunologia , Escherichia coli/imunologia , Vida Livre de Germes , Haptenos , Testes de Inibição da Hemaglutinação , Testes de Hemaglutinação , Humanos , Abscesso Hepático Amebiano/imunologia , Coelhos , Ratos , África do Sul , Staphylococcus/imunologia , Suécia , População BrancaRESUMO
Three major metabolites of propachlor were isolated from the excreta of germfree rats given 14C-labeled propachlor orally. In contrast, 11 urinary metabolites, six of which were 2-methylsulfonylacetanilides not present in excreta of germfree rats, were isolated from control rats given 14C-labeled propachlor orally. Enterohepatic circulation and microbial metabolism in the intestine were necessary for production of the methylsulfonyl-containing and other metabolites of propachlor in the conventional rat.
Assuntos
Acetanilidas/metabolismo , Vida Livre de Germes , Intestinos/microbiologia , Animais , Circulação Êntero-Hepática , Glutationa/metabolismo , Mucosa Intestinal/metabolismo , Fígado/metabolismo , Oxirredução , RatosRESUMO
Lateral epicondylitis is a common cause of elbow pain, and decreased microcirculation in extensor carpi radialis brevis (ECRB) has recently been suggested to contribute to the symptoms. The purpose of this pilot study was to investigate the treatment response after injection of botulinum toxin type A. Ten patients with unilateral epicondylitis and decreased intramuscular blood flow in ECRB participated. Handgrip, 2-pinch grip and muscle strength during radial deviation and dorsal extension of the wrist were recorded. Perceived pain during contraction was evaluated with the Visual Analogue Scale (VAS) and function in daily activities was assessed using the Disability of Arm, Shoulder and Hand instrument (DASH) and the Canadian Occupational Performance Measure instrument (COPM). Intramuscular blood flow was recorded by laser Doppler flowmetry, and microdialysis was used to analyze muscle metabolism. The difference in intramuscular blood flow between the control and the affected side had decreased 3 and 12 months after treatment (P=0.03). Lactate concentration at the 12-month follow-up had decreased (P=0.02); perceived pain was reduced and function in daily activities had improved. Injection of botulinum toxin is an alternative treatment for epicondylitis. Symptom relief may be due to enhanced microcirculation causing an aerobic metabolism.
Assuntos
Toxinas Botulínicas Tipo A/metabolismo , Músculo Esquelético/irrigação sanguínea , Músculo Esquelético/efeitos dos fármacos , Fármacos Neuromusculares/metabolismo , Fluxo Sanguíneo Regional/efeitos dos fármacos , Cotovelo de Tenista/tratamento farmacológico , Adulto , Toxinas Botulínicas Tipo A/farmacologia , Toxinas Botulínicas Tipo A/uso terapêutico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Força Muscular , Fármacos Neuromusculares/farmacologia , Fármacos Neuromusculares/uso terapêutico , Medição da Dor , Estudos Prospectivos , Cotovelo de Tenista/fisiopatologiaRESUMO
The dietary origin of the weak oestrogen equol (7-hydroxy-3-(4'-hydroxyphenyl)-chroman) present in human urine has been investigated using gas chromatography-mass spectrometry. Feeding experiments with different food constituents and monitoring the urinary excretion of equol revealed that soya food yields more than 0.1 mg urinary equol/g flour ingested. From this source the glucoside of daidzein (4',7-dihydroxyisoflavone) has been isolated and identified as a precursor of equol. Both equol and daidzein were characterized as monoglucuronide conjugates in human urine and the concentration of urinary equol exceeded the concentrations of the classical oestrogens by 100- to 1000-fold after ingestion of a single meal containing soya protein. The potential biological significance of this result is discussed.
Assuntos
Benzopiranos/urina , Cromanos/urina , Dieta , Glycine max/metabolismo , Adulto , Animais , Cromanos/análise , Equol , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Isoflavonas/análise , Masculino , Ratos , Ratos Endogâmicos , Glycine max/análiseRESUMO
The aim of this investigation was to study the influence of chenodeoxycholic acid administration on cholesterol and bile acid synthesis in germ-free rats. Seven rats were fed a basal diet and 2 groups of 4 rats received the same diet supplemented with 0.4 and 1% chenodeoxycholic acid, respectively. After 6 weeks, feces were collected in one 3- and one 4-day pool for analysis of cholesterol and bile acids. When the sampling period was finished, the rats were killed and the liver microsomal fractions isolated. The activities of HMG CoA reductase and cholesterol 7 alpha-hydroxylase were determined, the 7 alpha-hydroxylase by a mass fragmentographic method. The 2 dominating bile acids in the untreated rats were cholic acid and beta-muricholic acid. During treatment with chenodeoxycholic acid, 60--70% of this bile acid was converted into alpha- and beta-muricholic acid, indicating a high activity of the 6 beta-hydroxylase. The excretion of cholic acid was almost completely inhibited and the 7 alpha-hydroxylase activity was decreased ca 75% in the rats fed 1% chenodeoxycholic acid. The activity of the hepatic HMG CoA reductase as unchanged. The fecal excretion of cholesterol increased 2--3 times. An accumulation of cholesterol was seen in the rats treated with 1% chenodeoxycholic acid, which was probably a result of the decreased catabolism of cholesterol to bile acids.
Assuntos
Ácidos e Sais Biliares/metabolismo , Ácido Quenodesoxicólico/administração & dosagem , Colesterol/metabolismo , Vida Livre de Germes , Animais , Colesterol 7-alfa-Hidroxilase/metabolismo , Dieta , Fezes/análise , Hidroximetilglutaril-CoA Redutases/metabolismo , Fígado/metabolismo , RatosRESUMO
Regardless of diet (semi-synthetic or lab chow) or strain (AGUS or SD), germ-free rats have tryptic activity in their faeces, whereas conventional rats never do. The activity in faeces from germ-free AGUS rats was significantly higher than from SD rats.
Assuntos
Fezes/enzimologia , Vida Livre de Germes , Ratos Endogâmicos , Tripsina/análise , Animais , Dieta , Feminino , Masculino , Ratos , Especificidade da EspécieRESUMO
On comparing germ-free and conventional rats, inactivation of the tryptic activity was found to take place in the caecum of conventional adult rats only. A microbial intestinal inactivation of the tryptic activity was established in suckling conventional rats within 10 days after birth. At 3 weeks of age, suckling germ-free rats were found to have less faecal tryptic activity than their early-weaned littermates.
Assuntos
Vida Livre de Germes , Intestinos/microbiologia , Lactação/metabolismo , Ratos/metabolismo , Tripsina/metabolismo , Animais , Feminino , GravidezRESUMO
The present concept of the intestinal microflora has been reviewed with stressing the fact that it represents an entity with a weight comparable to one of the larger organs of the body. It is composed of about 500 different strains, many of which as yet have not been isolated because 95% of them are anaerobic and fastidious in their growth requirements. The intimate relationship of the flora to the surface epithelium of the intestines and the Lieberkühn's crypts are brought forward. The production of germfree animals is reviewed as an accounting of the most obvious differences between the germfree animal and its conventional counterparts, as far as gastroenterology is concerned. This includes the protective function of the normal intestinal flora, the caecum enlargement in germfree rodents, the influence of the flora on the metabolism of bilirubin, intestinal mucin, and pancreatic enzymes. Differences found in germfree animals due to the absence of the microbial flora related to the metabolism of fatty acids, bile acids, cholesterol and steroid hormones are also reviewed. The germfree animal has even proven to be an important research tool in such fields as carcinogenesis and studies of Crohn's disease and ulcerative colitis. There is need of a wider use of the germfree animal as a baseline in studies on the interference of the intestinal microbial flora with factors and conditions of great physiological and clinical importance.
Assuntos
Colo/microbiologia , Vida Livre de Germes , Anaerobiose , Animais , Ácidos e Sais Biliares/metabolismo , Bilirrubina/metabolismo , Carcinógenos , Colesterol/metabolismo , Quimotripsina/metabolismo , Colite Ulcerativa/etiologia , Neoplasias do Colo/induzido quimicamente , Doença de Crohn/etiologia , Ácidos Graxos/metabolismo , Humanos , Camundongos , Mucinas/metabolismo , Neoplasias Experimentais/induzido quimicamente , Elastase Pancreática/metabolismo , Ratos , Tripsina/metabolismoAssuntos
Tiazóis/metabolismo , Animais , Biotransformação , Vida Livre de Germes , Intestinos/microbiologia , RatosRESUMO
The purpose of this pilot study was to investigate intramuscular microcirculation in extensor carpi radialis brevis (ECRB) in patients with lateral epicondylitis. Ten patients with unilateral epicondylitis, mean duration of symptoms of 39 (12-96) months participated. The diagnosis was based on clinical examination and none was under treatment for the last 6 months. Isometric handgrip strength, 2-pinch grip strength and muscle strength during radial deviation and dorsal extension were determined. Functional perceived pain was evaluated by a modified behaviour rating scale and perceived pain during contraction by visual analogue scale. Intramuscular and skin blood flow was recorded by a laser-Doppler flowmetry system technique (LDF) during stable temperature condition. Intramuscular blood flow was significantly lower in the affected side, 22.7+/-9.8 perfusion units (PU), as compared with 35.2+/-11.9 PU in the control side (P=0.01). There was no difference in skin blood flow or temperature between the affected and the control side. A positive correlation was found between the duration of symptoms and the difference in intramuscular blood flow between the affected and the control arm (r=0.65, P=0.06). The present data indicate that decreased microcirculation and anaerobic metabolism in ECRB may contribute to the lateral epicondylitis symptoms.
Assuntos
Músculo Esquelético/irrigação sanguínea , Cotovelo de Tenista , Adulto , Feminino , Força da Mão , Humanos , Fluxometria por Laser-Doppler , Masculino , Pessoa de Meia-Idade , Cotovelo de Tenista/diagnóstico por imagem , UltrassonografiaRESUMO
Conventional (CONV) rats were fed by stomach tube for five days with either benzylpenicillin, ampicillin, tetracykline, oxitetracykline, neomycin, bacitracin + neomycin, kanamycin or colistin. On the 2nd-3rd day all the animals developed one or several of the following symptoms or characteristics typical for germfree (GF) rats: no coprostanol formation, no stercobilin production, a GF pattern after gel electrophoresis of fecal supernatant and proteolytic activity in the feces. Under the same conditions succinylsulfathiazole or metronidazole had much less pronounced effects than the antibiotics. When clofibrate, acetylsalicylic acid or ferrous sulphate were administered the effects were none or negligible. The GF characteristics persisted for several weeks after the end of the administration of the drugs. In some instances this was the case up to 7 weeks, when the animals were contaminated by anal route with a suspension of the cecum contents from intact CONV animals. On the 2nd day after this treatment the GF characteristics had disappeared.
Assuntos
Antibacterianos/farmacologia , Vida Livre de Germes , Intestinos/microbiologia , Ampicilina/farmacologia , Animais , Aspirina/farmacologia , Bacitracina/farmacologia , Pigmentos Biliares/biossíntese , Colestanol/biossíntese , Clofibrato/farmacologia , Colistina/farmacologia , Eritromicina/farmacologia , Fezes/microbiologia , Compostos Ferrosos/farmacologia , Canamicina/farmacologia , Masculino , Metronidazol/farmacologia , Neomicina/farmacologia , Oxitetraciclina/farmacologia , Penicilina G/farmacologia , Ratos , Sorbitol/farmacologia , Sulfatiazóis/farmacologia , Tetraciclina/farmacologiaRESUMO
Conventional Sprague-Dawley rats were fed by gastric tube for 5 days with either benzylpenicillin, ampicillin, doxycycline, or clindamycin. In contrast to the pretreatment period fibrinolytic activity and active and immunoreactive trypsin and immunoreactive elastase were present in fecal extracts after 4 days of antibiotic administration. This is consistent with findings in germfree rats and represents an alteration in the intestinal microflora. The germfree characteristics persisted until a suspension of cecal contents from normal rats was administered by enema on the 26th day. In fecal extracts from the clindamycin-treated rats considerable amounts of active and immunoreactive enzymes were, however, still found 10 days after the enema. A possible explanation is that clindamycin or a metabolite remains in the intestinal tract for a long time. The pathophysiological significance of this finding is unknown.
Assuntos
Antibacterianos/farmacologia , Quimotripsina , Intestinos/enzimologia , Elastase Pancreática , Tripsina , Ampicilina/farmacologia , Animais , Cromatografia em Gel , Quimotripsina/análise , Quimotripsina/imunologia , Clindamicina/farmacologia , Doxiciclina/farmacologia , Fezes/enzimologia , Masculino , Elastase Pancreática/análise , Elastase Pancreática/imunologia , Penicilina G/farmacologia , Ratos , Tripsina/análise , Tripsina/imunologiaRESUMO
Water-soluble intestinal mucins were investigated in germfree (GF), exgermfree (EXG) and conventional (CONV) rats and in GF and CONV mice. After agar gel electrophoresis, all GF animals had similar specific band patterns demonstrated by PAS and Toluidine Blue. These patterns, never seen in CONV animals, disappeared in GF animals infected either with intestinal contents from CONV rats or mono-infected with a mucin converting microorganism, labelled Peptostreptococcus N. The intestinal microflora seem to have a profound influence on the water-soluble mucins, and specific microorganisms appear to be involved in the conversion of these substances. Any CONV animal with a GF mucin pattern in the faeces must be considered to have a disturbance of the normal intestinal microflora.
Assuntos
Fezes/análise , Vida Livre de Germes , Intestinos/análise , Mucinas/análise , Animais , Infecções Bacterianas/microbiologia , Eletroforese em Gel de Ágar , Feminino , Intestinos/microbiologia , Masculino , Camundongos , Peptostreptococcus/metabolismo , Ratos , SolubilidadeRESUMO
No urobilins are formed from bilirubin in germ-free rats. To isolate and investigate the strains of intestinal microorganisms responsible for this transformation, a suitable test medium was adopted. The strength of the medium and a rather high initial pH were found to be of importance. In this medium, suspensions of rat faeces and a single strain, Cl. ramosum (G62), converted bilirubin to urobilins. Cultivations of Cl. ramosum (G62) together with E. coli significantly enhanced the conversion, whereas addition of 4 other bacterial strains was without the influence. The highest in vitro formation of the urobilins was about 10% of the bilirubin present. When the 6 strains investigated in vitro were established in EXG rats, the in vivo conversion of bilirubin to urobilins was found to be about 15%, compared to 70% in CONV rats.
Assuntos
Bilirrubina/metabolismo , Clostridium/metabolismo , Vida Livre de Germes , Urobilina/biossíntese , Animais , Meios de Cultura , Escherichia coli/metabolismo , Técnicas In Vitro , Ratos , Ratos Endogâmicos/microbiologiaRESUMO
Conventional Sprague-Dawley rats were treated with clindamycin, 40 mg/kg/day and 0.04 mg/kg/day, for 5 days. At a dose of 40 mg/kg/day, microflora-associated characteristics (MACs), such as shape, color, and consistency of feces, proteolytic activity, electrophoretic pattern, and cholesterol and bilirubin metabolism were transformed into values like those found in germfree rats: germfree animal characteristics (GACs). The effect on the proteolytic activity lasted longest. It did not disappear until one or two enemas with cecal contents from intact conventional rats were administered. At a dose of 0.04 mg/kg/day, effects on the proteolytic activity and cholesterol metabolism were seen. With the exception of one rat, the effect on proteolytic activity did not disappear until one or two enemas were given. The results indicate that clindamycin, even in very small daily doses, has a profound and long-lasting influence on many intestinal MACs in rats.
Assuntos
Bactérias/efeitos dos fármacos , Clindamicina/farmacologia , Intestinos/microbiologia , Animais , Fezes/microbiologia , Masculino , Ratos , Ratos EndogâmicosRESUMO
Two electrophoretically distinct trypsins and chymotrypsins and an elastolytic enzyme were isolated from rat pancreatic juice. Rabbit antisera against these enzymes were produced, and with an immunochemical technique the trypsins, chymotrypsins, and elastase were studied in the intestinal contents of conventional and germfree rats. In both types of rat the anionic trypsin and chymotrypsin were the most abundant and found in higher concentrations in the distal than in the proximal small intestine. The cecal and fecal concentrations of anionic trypsin were markedly higher in the germfree rat when compared to the conventional rat. Cymotrypsin was undetectable in the large intestine of either the conventional or germfree rat when this technique was used. Immunoreactive elastase was found in greater amounts in the distal small intestine, and high concentrations were demonstrated in the cecal contents and feces of the germfree rat. In contrast, no immunoreactive elastase was detected in the large intestine of the conventional rat. Gel filtration indicated that the immunoreactive anionic trypsin and elastase found in fecal extracts were of about the same molecular size as the native enzymes. The findings suggest that the intestinal microflora is instrumental in the inactivation and degradation of pancreatic trypsin and elastase but not chymotrypsin.
Assuntos
Quimotripsina/análise , Intestinos/enzimologia , Elastase Pancreática/análise , Tripsina/análise , Animais , Cromatografia em Gel , Quimotripsina/imunologia , Fezes/enzimologia , Vida Livre de Germes , Técnicas Imunológicas , Elastase Pancreática/imunologia , Suco Pancreático/enzimologia , Ratos , Tripsina/imunologiaRESUMO
Trypsin, chymotrypsin, and elastase were measured in the intestinal contents of germfree and conventional laboratory rats. Active trypsin was highest in the lower part of the small intestine and fell abruptly in the cecum. Trypsin was present in the feces of germfree rats but not of conventional laboratory rats. Electrophoresis of supernatant fluids from intestinal contents, and functional assays with specific synthetic substrates confirmed these results. The concentrations of active elastase were very high in the upper small intestine but low elsewhere. Active elastase was present in contents from the large intestine of germfree, but not conventional laboratory rats. These results suggest that normal microbiological inhabitants of the intestinal tract of rats play a role in the inactivation of pancreatic enzymes.