RESUMEN
AIM: To evaluate the influence of root canal form on the sealing ability of two root canal sealers. METHODOLOGY: Twenty radiographically confirmed straight and 20 curved root canals were prepared with a stepback hand filing technique. Root canal aberrations created during preparation were determined by the use of double exposure radiographic technique. The prepared canals were filled with lateral condensation of gutta-percha and one or other of two root canal sealers (Pulp Canal Sealer and Sealapex). Leakage along the apical 10 mm of roots was measured with a fluid transport model at 1, 3, 6, 9 and 12-month intervals. RESULTS: There were no statistically significant differences between straight and curved root canals (P > 0.05) for prevalence of root canal transportation. The prevalence of apical transportation was 80% in the straight and 85% in the curved root canals. A complete seal was more frequently observed in straight canals compared with curved canals. Utilizing the pi* index, analysis showed the filling with Sealapex allowed more leakage than Pulp Canal Sealer at 1 year. CONCLUSION: Under the conditions of the study, root canal form influenced short-term sealing ability. In the long-term the seal was affected by the sealer rather than root canal form.
Asunto(s)
Filtración Dental , Cavidad Pulpar/anatomía & histología , Materiales de Obturación del Conducto Radicular , Análisis de Varianza , Hidróxido de Calcio , Humanos , Preparación del Conducto Radicular/instrumentación , Salicilatos , Estadísticas no Paramétricas , Cemento de Óxido de Zinc-EugenolRESUMEN
A quantitative longitudinal study was carried out to compare the amount of microleakage using two different types of root canal sealers on straight and curved root canals. Microleakage was measured just after the setting of the sealer, one month and three months after that using fluid transport model. One month follow up results showed that in straight root canals both the ZOE containing Pulp Canal Sealer and the calciumhydroxide containing Sealapex sealers produced similar microleakage values to that values measured just after the setting of materials. One month follow up measurement resulted higher values in curved canals comparing to the straight ones in case of both the tested materials. Three months follow up measurement demonstrated worse results in the Sealapex groups than the Pulp Canal Sealer groups. With loglinear statistical analysis the following interactions were proved between variables: root canal form/microleakage, root canal form/time laps, and material/microleakage/time laps. Results of the present study are in good agreement with conclusions of recently published other studies, namely calciumhydroxide containing sealers resulted in higher microleakages than ZOE containing sealers.
Asunto(s)
Hidróxido de Calcio , Materiales de Obturación del Conducto Radicular/normas , Cemento de Óxido de Zinc-Eugenol , Humanos , Hungría , Estudios LongitudinalesRESUMEN
In studies to determine whether Saccharomyces cerevisiae produced estrogens, the organism was grown in culture media prepared using distilled water autoclaved in polycarbonate flasks. The yeast-conditioned media showed the presence of a substance that competed with [3H]estradiol for binding to estrogen receptors (ER) from rat uterus. However, it soon became clear that the estrogenic substance in the conditioned media was not a product of the yeast grown in culture, but was leached out of the polycarbonate flasks during the autoclaving procedure. [3H]Estradiol displacement activity was monitored by ER RRA, and the active substance was purified from autoclaved medium using a series of HPLC steps. The final purified product was identified as bisphenol-A (BPA) by nuclear magnetic resonance spectroscopy and mass spectrometry. BPA could also be identified in distilled water autoclaved in polycarbonate flasks without the requirement of either the organism or the constituents of the culture medium. Authentic BPA was active in competitive RRAs, demonstrating an affinity approximately 1:2000 that of estradiol for ER. In functional assays, BPA (10-25 nM) induced progesterone receptors in cultured human mammary cancer cells (MCF-7) at a potency of approximately 1:5000 compared to that of estradiol. The BPA effect on PR induction was blocked by tamoxifen. In addition, BPA (25 nM) increased the rate of proliferation of MCF-7 cells assessed by [3H]thymidine incorporation. Thus, BPA exhibited estrogenic activity by both RRA and two functional bioresponse assays. Finally, MCF-7 cells grown in media prepared with water autoclaved in polycarbonate exhibited higher progesterone receptor levels than cells.grown in media prepared with water autoclaved in glass, suggesting an estrogenic effect of the water autoclaved in polycarbonate. Our findings raise the possibility that unsuspected estrogenic activity in the form of BPA may have an impact on experiments employing media autoclaved in polycarbonate flasks. It remains to be determined whether BPA derived from consumer products manufactured from polycarbonate could significantly contribute to the pool of estrogenic substances in the environment.
Asunto(s)
Estrógenos/farmacología , Fenoles/farmacología , Cemento de Policarboxilato , Esterilización/métodos , Animales , Compuestos de Bencidrilo , Cromatografía Líquida de Alta Presión , Medios de Cultivo , Equipos y Suministros , Estrógenos/metabolismo , Femenino , Humanos , Laboratorios , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Fenoles/análisis , Fenoles/química , Cemento de Policarboxilato/química , Receptores de Estrógenos/metabolismo , Receptores de Estrógenos/fisiología , Receptores de Progesterona/metabolismo , Saccharomyces cerevisiae/metabolismo , Células Tumorales Cultivadas , Útero/metabolismo , Agua/química , Agua/farmacologíaRESUMEN
1. The biotransformation of nafimidone, an imidazole-substituted anticonvulsant, has been studied by characterization of urinary metabolites in dogs, cynomolgus monkeys, baboons and man. 2. The biotransformation of nafimidone in these laboratory animals and man is initially very similar, in each case proceeding by reduction to the aliphatic alcohol metabolite, nafimidone alcohol or 1-[2-hydroxy-2-(2-naphthyl)ethyl]imidazole. 3. Further transformation of this metabolite involves oxidation in the naphthyl and imidazole functions, and/or conjugation. 4. The dog differs from the higher primates in that no metabolic modification of the naphthyl group takes place, the major metabolite in the dog being the O-beta-glucuronide of nafimidone alcohol. 5. In higher primates and man two isomers involving dihydroxylation in the naphthyl ring--1-[2-hydroxy-2-(5,6- or 7,8-dihydroxydihydro-2-naphthyl)ethyl]imidazole--were tentatively identified. These species alone showed evidence of an imidazole linked N-glucuronide of nafimidone alcohol. 6. The possible occurrence of stereoselective metabolism by the introduction of a chiral centre at C-9 in nafimidone alcohol was indicated in human urine by the presence of both epimers of the O-beta-glucuronide of nafimidone alcohol in a 2:1 ratio.
Asunto(s)
Imidazoles/metabolismo , Nafazolina/metabolismo , Adulto , Animales , Anticonvulsivantes/metabolismo , Anticonvulsivantes/orina , Perros , Haplorrinos , Humanos , Masculino , Espectrometría de Masas , Persona de Mediana Edad , Nafazolina/análogos & derivados , Nafazolina/orinaRESUMEN
It has been shown by us and others that progesterone inhibits the growth of Trichophyton mentagrophytes and that the organism escapes from this inhibition over time. We report here studies which show that escape from growth inhibition is related to the enzymatic transformation of progesterone to polar metabolites. Isolation and identification of the progesterone metabolites confirm the production of 15 alpha-hydroxyprogesterone. In addition, three other metabolites were isolated. Two of these were determined to be 1-dehydroprogesterone and 11 alpha-hydroxyprogesterone. The third metabolite was a 1-dehydro-hydroxyprogesterone, but the location of the hydroxyl group could not be determined unequivocally. Studies using authentic 15 alpha-hydroxyprogesterone, 1-dehydroprogesterone, and 11 alpha-hydroxyprogesterone reveal that these derivatives are significantly less inhibitory to the growth of T. mentagrophytes than progesterone. Pretreatment of organisms with progesterone augments the rate of metabolism and enhances escape. We have described previously a progesterone-binding protein (PBP) in cytoplasmic extracts of T. mentagrophytes and hypothesized that progesterone mediates growth inhibition by binding to the PBP of this organism. The relative binding affinity that progesterone and its metabolites display for PBP correlates with the relative growth inhibitory potency of these compounds. These results suggest that metabolism of progesterone to more polar and less inhibitory compounds, which exhibit lower affinity for PBP, is the mechanism of escape from progesterone-mediated inhibition of growth in this organism.
Asunto(s)
Progesterona/metabolismo , Trichophyton/crecimiento & desarrollo , Unión Competitiva , Cromatografía Líquida de Alta Presión , Hidroxiprogesteronas/metabolismo , Hidroxiprogesteronas/farmacología , Cinética , Espectrometría de Masas , Progesterona/análogos & derivados , Progesterona/farmacología , Globulina de Unión a Progesterona/metabolismo , Trichophyton/efectos de los fármacos , Trichophyton/metabolismoRESUMEN
Ketorolac tromethamine (KT), a potent non-narcotic analgesic, with cyclooxygenase inhibitory activity, was administered (14C-labeled and unlabeled) intravenously (iv), orally (po), and intramuscularly (im) in solution to humans, cynomolgus monkeys, rabbits, rats, and mice. KT was absorbed rapidly (Tmax less than 1.0 hr) and efficiently (greater than 87%) following po and im doses in all species. The plasma half-life of ketorolac (K) ranged from 1.1 hr (rabbits) to 6.0 hr (humans). The protein binding of K ranged from 72.0% (mouse) to 99.2% (humans). Linear pharmacokinetics of K was observed in the mouse after single oral doses of KT ranging from 0.25 to 16 mg/kg. Radioactivity was excreted predominantly into urine, ranging from 78.9% (mouse) to 102% (monkey) following iv doses. The dose was excreted into urine primarily as K conjugates, K, and p-hydroxy-K in humans. The monkey was similar to humans with respect to kinetics, but did not form the p-hydroxy metabolite. The rabbit was unusual in that it exhibited substantial presystemic metabolism (50%). The rat excreted a much higher percentage of radioactivity into the feces and formed an additional unidentified metabolite. The most comparable species with respect to humans metabolically was the mouse. The metabolism and excretion of K was similar following iv, po, and im doses within each species studied.
Asunto(s)
Pirroles/farmacocinética , Tolmetina/farmacocinética , Trometamina/farmacocinética , Administración Oral , Adulto , Animales , Cromatografía Líquida de Alta Presión , Combinación de Medicamentos/administración & dosificación , Combinación de Medicamentos/farmacocinética , Femenino , Humanos , Inyecciones Intramusculares , Inyecciones Intravenosas , Ketorolaco Trometamina , Macaca fascicularis , Masculino , Ratones , Conejos , Ratas , Ratas Endogámicas , Distribución Tisular , Tolmetina/administración & dosificación , Tolmetina/análogos & derivados , Trometamina/administración & dosificaciónRESUMEN
We have previously shown the presence of 17 beta-estradiol in extracts of commercially prepared Saccharomyces cerevisiae ss well as the production of estradiol by yeast grown in the laboratory. In our current study, yeast grown in a chemically defined medium synthesized estradiol in only small amounts, (less than 500 pg/liter). We have analyzed a variety of media commonly used for growing yeast and found that substantial estradiol production (greater than 5 ng/liter) was obtained when yeast were grown in medium supplemented with Bacto-peptone. The peptone was shown to contain significant amounts of estrone, and the results of the experiments establish a precursor-product relationship where estrone from the medium is metabolized to estradiol by S. cerevisiae. Studies with added [3H]estrone demonstrated rapid conversion into [3H]estradiol and a 3H-labeled nonpolar estrogen derivative. The commercially obtained yeast used previously had been grown in a molasses medium. We demonstrate here that the molasses medium contains substantial amounts of estrone and estradiol. We conclude that the conversion of estrone in a culture medium to estradiol in laboratory grown yeast and estrone and estradiol present in the commercially grown yeast medium account for the majority of estradiol found in yeast.
Asunto(s)
Estrógenos/metabolismo , Saccharomyces cerevisiae/metabolismo , Cromatografía Líquida de Alta Presión , Medios de Cultivo , Estradiol/análisis , Estrona/análisis , Melaza , Peptonas , Radioinmunoensayo , Factores de TiempoRESUMEN
Prostaglandin D2 spontaneously decomposes at physiological pH and temperature to 9-deoxy-delta 9-PGD2 (designated PGJ2). We developed a TLC procedure for the isolation of PGJ2 which was identified by both proton-NMR and mass spectrometry. Freshly prepared PGJ2 was active in inhibiting aggregation induced by ADP in citrated human platelet rich plasma. As reported by Fukushima et al. (1). PGJ2 was less active (x 0.1-0.25) than PGD2 as an inhibitor. Concentrations of PGJ2 that markedly inhibited aggregation of human platelets were generally incapable of inhibiting aggregation of rat or guinea pig platelets. Using a heterologous system of human platelets mixed with guinea pig plasma samples (2), it was shown that the ability of PGJ2 to inhibit platelet aggregation was lost immediately following intravenous injection in anesthetized guinea pigs. This apparent rapid uptake and/or degradation of PGJ2 might also explain why PGJ2 had no effect on blood pressure of anesthetized guinea pigs. PGJ2 was potent in inhibiting proliferation of cultured vascular smooth muscle cells, mouse melanoma cells and mouse fibroblasts. Less potent anti-proliferative effects were seen with two other degradation products of PGD2, one of which was the delta 12 metabolite reported (3,4) to be formed from PGJ2 in a reaction catalyzed by serum albumin.
Asunto(s)
Prostaglandinas D/análisis , Adenosina Difosfato/farmacología , Animales , Presión Sanguínea/efectos de los fármacos , División Celular/efectos de los fármacos , Línea Celular , Células Cultivadas , Dinoprostona , Cobayas , Humanos , Espectroscopía de Resonancia Magnética , Masculino , Espectrometría de Masas , Agregación Plaquetaria/efectos de los fármacos , Prostaglandina D2 , Prostaglandinas D/farmacología , Prostaglandinas E/farmacologíaRESUMEN
Saccharomyces cerevisiae possesses a high-affinity estrogen binding protein and an endogenous ligand that displaces [3H]estradiol from both the yeast binding protein and mammalian estrogen receptors. Semipurified preparations of this ligand have been shown to exhibit estrogenic activity in mammalian systems. We now describe the purification procedure and ultimate identification of the estrogenic substance in extracts of S. cerevisiae as 17 beta-estradiol. Organic solvent extracts of commercially obtained dried yeast were sequentially chromatographed on silica gel columns and then subjected to a series of reversed phase HPLC fractionations. Active ligand was monitored by [3H]estradiol displacement in a rat uterine cytosol assay. After seven chromatography steps, the purified and highly active ligand exhibited a single peak with retention times identical to those of 17 beta-estradiol on both HPLC and GC. The yeast material was identified as 17 beta-estradiol by its UV absorbance and mass spectrometric fragmentation pattern. In addition, radioimmunoassay confirmed the presence of approximately the same mass of 17 beta-estradiol (approximately equal to 800 ng/1.5 kg of yeast) as estimated both by a competitive binding assay with estrogen receptor and by mass spectrometry. Extraneous contamination by estradiol was excluded by repeat experiments with different batches of starting material and demonstration of estradiol by RIA in conditioned medium and cell pellets of laboratory-grown S. cerevisiae whereas non-conditioned medium did not possess the steroid. We conclude that 17 beta-estradiol is a yeast product.
Asunto(s)
Estradiol/aislamiento & purificación , Saccharomyces cerevisiae/análisis , Estradiol/inmunología , RadioinmunoensayoAsunto(s)
Antiinflamatorios/metabolismo , Fluocinolona Acetonida/análogos & derivados , Administración Tópica , Radioisótopos de Carbono , Cromatografía Líquida de Alta Presión , Cromatografía en Capa Delgada , Fluocinolona Acetonida/metabolismo , Humanos , Masculino , Espectrometría de Masas , Técnica de Dilución de RadioisótoposRESUMEN
Flunisolide (6 alpha-fluoro-11 beta, 16 alpha, 17 alpha, 21-tetrahydroxypregna-1,4-diene-3,20-dione-16,17-acetonide) was converted to 6 beta,- 11 beta, 16 alpha, 21-pentahydroxypregna-1,4-diene-3,20-dione-16,17-acetonide (6 beta-OH metabolite) by mouse liver microsomes, but no activity was observed with mouse lung, intestine or kidney microsomes. Two additional metabolites of flunisolide also formed by mouse hepatic microsomes were identified by mass spectral analysis to be 11 beta, 16 alpha, 17 alpha,21-tetrahydroxypregna-1,4-diene-3,6,20-trione-16,17-acetonide (6-keto metabolite) and delta 6-flunisolide. The formation of all three metabolites required NADPH, was inhibited by carbon monoxide and was stimulated by pretreating mice with phenobarbital. A time-couse study suggested the 6-keto metabolite was an intermediate in the formation of the 6 beta-OH metabolite. When added to microsomes, the 6-keto metabolite was converted to the 6 beta-OH metabolite by a carbon monoxide-insensitive enzyme. Our results suggest the conversion of flunisolide to the 6 beta-OH metabolite is catalyzed by a multi-enzyme pathway via a stable intermediate, the 6-keto metabolite. The initial reaction which leads to the formation of the 6-keto metabolite is catalyzed by a cytochrome P-450-mediated microsomal monoxygenase(s), but the reduction of the 6-keto metabolite to the 6 beta-OH metabolite is cytochrome P-450-independent.
Asunto(s)
Fluocinolona Acetonida/análogos & derivados , Animales , Biotransformación , Femenino , Fluocinolona Acetonida/metabolismo , Flúor/metabolismo , Técnicas In Vitro , Espectrometría de Masas , Ratones , Microsomas Hepáticos/metabolismo , Oxidación-Reducción , Fenobarbital/farmacologíaRESUMEN
1. The bile salts of three frog species of the genus Ptychadena and of Rana catesbeiana have been compared with those of their tadpoles. For R. catesbeiana comparison was made of the bile salts in at least ten of the recognized stages of tadpole metamorphosis. 2. In all cases, adult bile salts were more complex than those of the tadpoles. 3. In R. catesbeiana after stage 18, 26-deoxy-5 alpha-ranol was hydroxylated to form 5 alpha-ranol (27-nor-5 alpha-cholestane-3 alpha, 7 alpha, 12 alpha, 24 xi, 26-pentol) and at least two other bile alcohols appeared in solvolysed bile salts. 4. Tadpole bile salts were not found to be biochemically more primitive than those of fully metamorphosed frogs; in some, but not all, cases tadpole bile alcohols could be regarded as biochemical precursors of those in the adult frogs. 5. Detailed evidence for the structure of the bile salts from mass-spectral fragmentation patterns has been deposited as Supplementary Publication SUP 50097 (2 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1978) 169, 5.
Asunto(s)
Ácidos y Sales Biliares/metabolismo , Metamorfosis Biológica , Rana catesbeiana/metabolismo , Animales , Anuros , Cromatografía de Gases y Espectrometría de Masas , Larva/metabolismo , Rana catesbeiana/crecimiento & desarrolloRESUMEN
Flunisolide (6 alpha-fluoro-11 beta, 16 alpha, 17 alpha, 21-tetrahydroxypregna-1,4-diene-3,20-dione 16,17-acetonide) administered as a single iv or oral dose to rats, mice, dogs, rhesus monkeys, and cynomolgus monkeys had a plasma t 1/2 of 1-3.5 hr and was eliminated mainly via the bile. After iv administration of 14C-labeled flunisolide, radioactivity was widely distributed into tissues and organs. The apparent volume of distribution of flunisolide in these five species was 3.0-8.0 liters/kg. A major metabolite isolated from rhesus monkey urine was shown to be 6 beta, 11 beta, 16 alpha, 17 alpha, 21-pentahydroxypregna-1,4-diene-3,20-dione 16,17-acetonide (6 beta-OH metabolite). Free 6 beta-OH metabolite was a major radioactive entity detected in urine of all species given radiolabeled flunisolide, whereas flunisolide conjugated with glucuronic acid and/or sulfate was a major metabolite detected in the bile of rats, dogs, and cynomolgus monkeys. Following the oral administration of radiolabeled flunisolide, radioactivity was rapidly and efficiently absorbed in all species, but in the rhesus and cynomolgus monkeys most of the plasma radioactivity was due to the 6 beta-OH metabolite and to water-soluble conjugates, suggesting extensive first-pass metabolism of flunisolide.
Asunto(s)
Fluocinolona Acetonida/análogos & derivados , Animales , Bilis/metabolismo , Biotransformación , Perros , Femenino , Fluocinolona Acetonida/metabolismo , Haplorrinos , Absorción Intestinal , Cinética , Macaca fascicularis , Macaca mulatta , Masculino , Ratones , Ratas , Especificidad de la EspecieRESUMEN
1. Bile salts of the green turtle Chelonia mydas (L.) were analysed as completely as possible. 2. They consist of taurine conjugates of 3 alpha, 7 alpha, 12 alpha, 22 xi-tetrahydroxy-5 beta-cholestan-26-oic acid (tetrahydroxysterocholanic acid) and 3 alpha 12 alpha, 22 xi-trihydroxy-5 beta-cholestan-26-oic acid, with minor amounts of 3 alpha, 7 alpha, 12 alpha-trihydroxy-5beta-cholan-24-oic acid (cholic acid), 3alpha, 12 alpha-dihydroxy-5beta-cholan-24-oic acid (deoxycholic acid) and possibly other bile acids. 3. Cholic acid and deoxycholic acid represent the first known examples of bile acids common to chelonians and other animal forms: they may indicate independent evolution in chelonians to C24 bile acids. 4. The discovery of a 7-deoxy C27 bile acid is the first evidence that C27 bile acids or their conjugates have an enterohepatic circulation.
Asunto(s)
Ácidos y Sales Biliares/análisis , Tortugas/metabolismo , AnimalesRESUMEN
1. Bile salts of the coelacanth Latimeria chalumnae Smith (five specimens) and of the three living genera of lungfish (Dipnoi) were examined as completely as possible and compared. 2. The small 'bile acid' fractions include no more than traces of well-known C27 or C24 acids (free or conjugated) and the functioning bile salts must be regarded as alcohol sulphates. 3. Comparison of the alcohols suggest that (a) Latimeria stands biochemically outside the animal group which includes the Dipnoi, (b) Protopterus and Lepidosiren are more closely related to one another than either is to Neoceratodus, (c) all four primitive osteiychtheans have some amphibian affinities, (d) there are affinities between Latimeria and Dipnoi and ostariophysan families (especially Cyprinidae and Catostomidae) and (e) there are biochemical links between Dipnoi and lampreys.
Asunto(s)
Ácidos y Sales Biliares/análisis , Peces/metabolismo , Anfibios , AnimalesRESUMEN
dl-9-Oxo-13 (cis and trans)-prostenoic acids were converted in high yields into their 18- and 19-hydroxy derivatives by cultures of Microascus trigonosporus. The structure elucidation of the microbial products is described.
Asunto(s)
Ascomicetos/metabolismo , Ácidos Grasos Insaturados/metabolismo , Fenómenos Químicos , Química , HidroxilaciónRESUMEN
Deuterium- and tritium-labeled chenodeoxycholic acid and lithocholic were prepared by catalytic reduction of their respective delta11 derivatives. Structures of the intermediates and their isotopic purity were verified by chemical ionization and electron impact mass spectrometry and by nuclear magnetic resonance spectroscopy. Experimental conditions for reductive deuteration were defined which gave complete reduction of the olefin and a product of high isotopic purity. Conditions for optimal tritiation were developed with which little exchange of protons with the solvent occurred; the product had high specific activity. To test biological stability of the label, the 3H-labeled chenodeoxycholic acid was administered simultaneously with 14C-labeled chenodeoxycholic acid to two healthy subjects and the 3H/14C ratio in bile was determined daily for several days. The ratio remained identical to that administered, suggesting that the 11,12-3Hlabel in chenodeoxycholic acid is stable during enterohepatic cycling and can be used for valid estimates fo bile acid kinetics in many by the isotope dilution technique.
Asunto(s)
Ácido Quenodesoxicólico , Ácidos Cólicos , Deuterio , Ácido Litocólico , Tritio , Ácido Quenodesoxicólico/síntesis química , Ácido Quenodesoxicólico/metabolismo , Colanos , Colenos , Ácidos Cólicos/síntesis química , Estabilidad de Medicamentos , Heces , Humanos , Marcaje Isotópico , Ácido Litocólico/síntesis química , Ácido Litocólico/metabolismo , Espectrometría de Masas , Oxidación-ReducciónRESUMEN
1. Methods have been developed for the isolation and identification of small amounts of bile salts and of bile acids and alcohols obtained by solvolysis. These methods involve preparative and analytical t.l.c., purification on columns of protonated Al(2)O(3) and Sephadex LH-20 and also g.l.c.-mass spectroscopy of solvolysis products. 2. Application to 29 species of frogs and toads has confirmed the constancy of bile salt patterns in a single species, including colour phases in two instances, and has revealed great variations between different species in some genera (e.g. Rana, Ptychadena) and little difference between widely distributed species in others (e.g. Bufo). 3. Taxonomic deductions should be made with caution and with regard to the physiological significance of the biochemical character considered. The molecular differences found might be interpreted as indicating variations in the rate of evolution.