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Biomedical subjects

A M Batlle

Publications and source records attributed to A M Batlle.

At least 55 records · Page 3Linked to original sources
PubMed1992

Uroporphyrinogen decarboxylase from mouse mammary carcinoma and liver of normal and tumor-bearing mouse.

1. URO-D was investigated in crude extracts from mouse mammary carcinoma, normal mouse (NM) liver and tumor-bearing mouse (TBM) liver. 2. URO-D from TBM liver and tumor appears to be more sensitive to increasing concentrations of UROgen than the NM liver enzyme. 3. In tumor the rate-limiting step seems to be the decarboxylation of the first carboxyl group, but this was not so clear for the NM and the TBM liver URO-D. 4. URO-D activity was enhanced when incubated at higher temperatures in the presence of its substrate, suggesting that UROgen might afford some protection of the enzyme against heat inactivation. 5. The optimum pH for all three sources is around 7.0.

Animals↗
PubMed1992

Porphyrinogenic properties of the anesthetic enflurane.

1. The effect of enflurane, a volatile anesthetic, on heme metabolism was studied. Different doses (0.5-6.0 ml/kg) of this anesthetic were administered i.p. to mice and animals sacrificed at different times after administration (5-240 min). 2. The dose of 2 ml/kg was chosen as the optimum anesthetic dose producing more alterations in the heme pathway. 3. ALA-S was significantly induced at earlier times of anesthesia. 4. Blood PBGase and deaminase was greatly reduced. 5. This diminution coupled with ALA-S induction are in accordance with the known biochemical changes occurring in acute intermittent porphyria and include enflurane in the list of porphyrinogenic drugs, the use of which is not recommended for the management of anesthesia in porphyric patients.

5-Aminolevulinate Synthetase↗
PubMed1991

Different porphobilinogen-deaminase forms in wild and mutant strains of Saccharomyces cerevisiae. A possible correlation with its segregants behaviour.

1. Different porphobilinogen-deaminase (PBG-D) enzyme forms were found for D 27 and D 27/C6 (HEM R+) strains of Saccharomyces cerevisiae. 2. PBG-D was partially purified and chromatographed on Sephadex G-100 in either the presence or absence of a protease inhibitor. For D 27 only one active peak was observed while for D 27/C6 strain two active peaks were found. 3. A correlation between this differential behaviour and the presence of HEM R+ gene was looked for employing two segregants of one tetrad from D 27 and D 27/C6 mating.

Ammonia-Lyases↗
PubMed1991

Mouse mammary carcinoma porphobilinogenase and hydroxymethylbilane synthetase.

1. Porphobilinogenase (PBGase) and hydroxymethylbilane synthetase (HMB-S) were investigated in crude extracts from mouse mammary carcinoma, normal mouse liver and tumor bearing mouse liver. 2. A Michaelis-Menten kinetics for both enzymes in either source was observed. Km values of 87 to 108 microM, Vmax of 1.57-1.83 nmol porphyrins/2 hr and a Hill coefficient of n = 1 were obtained for PBGase and Km values of 13 to 19 microM and Vmax of 2.6-4.8 were obtained for HMB-S. 3. Porphyrin synthesis was linear up to 180 min of incubation in all cases for PBGase and HMB-S, and greatly increased with higher incubation temperature being maximal at 60 degrees C and nil at 70 degrees C, optimal temperature was 37 degrees C for either enzyme in either source. 4. Uroporphyrinogen III synthetase was heat inactivated while HMB-S was a heat stable enzyme. Optimum pH was 8.2 for PBGase and HMB-S in either tissue.

Ammonia-Lyases↗
PubMed1991

The effect of griseofulvin on the heme pathway--II. An exhaustive analysis during short and long-term challenge.

1. A clear biphasic response of the enzyme activities as a function of intoxication time due to the topical cutaneous griseofulvin treatment was observed. 2. The initial acute induction of ALA-S activity would be due to depletion of free heme in the regulatory pool caused by cytochrome P 450 destruction. 3. The second induction peak, would be due to less heme formation, secondary to the ferrochelatase inhibition, as expected for the erythropoietic protoporphyria model. 4. The biphasic response of hepatic ALA-D and PBGase activities would be related to ALA-S activity changes and the subsequent augmented available substrates. 5. Endogenous liver porphyrin distribution in cytosolic, mitochondrial and nuclear fractions was investigated. 6. The in vitro biosynthesis of porphyrins confirmed both the biphasic model and the hepatic porphyrins subcellular distribution. 7. Two mechanisms to explain the action of griseofulvin at shorter and longer times of intoxication are proposed.

5-Aminolevulinate Synthetase↗
PubMed1991

Further evidence on the photodynamic and the novel non-photodynamic inactivation of uroporphyrinogen decarboxylase by uroporphyrin I.

The action of uroporphyrin I (URO I) on the activity of red cell uroporphyrinogen decarboxylase (URO-D) in the dark and under UV light was studied. Light-dependent-and light-independent inactivation was observed. Both effects increased at increasing concentrations of URO I, the former reached its maximum at 150 microM of sensitizer. At 100 microM of URO I, both light and dark inactivation were temperature dependent amounting to about 50% at 30-37 degrees C. The velocity of dark inactivation increased with increasing temperature in the range of 0 to 45 degrees C. Photoinactivation can be ascribed to primary oxidation of essential amino acids, very likely histidyl residues, followed by secondary inter or intrapeptide cross-linking. Dark inactivation could be the result of both oxidation and cross-linking (although to a less degree than that produced by light) and also direct inhibition of the enzyme by induced conformational changes at its active site through binding of the porphyrin to the protein. When the action of URO I was tested on partially purified URO-D, the enzyme appeared to be more susceptible to the dark than to the light effect.

Darkness↗
PubMed1991

Involvement of free and enzyme-bound intermediates in the reaction mechanism catalyzed by the bovine liver immobilized porphobilinogen deaminase. Proof that they are substrates for cosynthetase in uroporphyrinogen III biosynthesis.

The detection and accumulation of tetrapyrrole intermediates synthesized by the action of bovine liver porphobilinogen deaminase immobilized to Sepharose 4B is reported. Employing Sepharose-deaminase preparations, two phases in uroporphyrinogen I synthesis as a function of time were observed, suggesting the accumulation of free and enzyme-bound intermediates, the concentration and distribution of which were time dependent. The deaminase-bound intermediate behaves as a substrate in uroporphyrinogen I synthesis whereas the free intermediates produce enzyme inhibition. The tetrapyrrole intermediate bound to the Sepharose-enzyme is removed from the protein by the binding of porphobilinogen. Free as well as enzyme-bound intermediates are shown to be substrates for cosynthetase with formation of 80% uroporphyrinogen III.

Animals↗
PubMed1990

Heme regulation in mouse mammary carcinoma and liver of tumor bearing mice--I. Effect of allyl-isopropylacetamide and veronal on delta-aminolevulinate synthetase, cytochrome P-450 and cytochrome oxidase.

1. Basal levels and allyl-isopropylacetamide (AIA) or veronal induced levels of delta-aminolevulinate synthetase (ALA-S), cytochrome P-450 (cyt P-450) and cytochrome oxidase were determined in tumor (T) and liver of both normal mice (NM) and T bearing mice (TBM). 2. Basal levels of ALA-S were nearly the same in either source. The amount of cyt P-450 was lower in TBM liver than in NM liver, and no detectable in T. While the basal activity of cytochrome oxidase in TBM liver and T were higher than those of NM liver. 3. In AIA intoxicated animals there was a lower induction of ALA-S in liver of TBM than in NM liver. There was no induction in T ALA-S. The loss of cyt P-450 was less in TBM liver when compared with NM liver. 4. The induction level of cyt P-450 after veronal administration was nearly the same in liver of both TBM and NM. 5. We conclude that lower induction of liver ALA-S activity in TBM liver is due to correspondingly lower drug metabolism ability of TBM liver. Otherwise our results suggest that the control mechanism operating in T and probably in its original tissue are different from those described for normal liver.

5-Aminolevulinate Synthetase↗
PubMed1990

Mouse mammary carcinoma delta-aminolevulinate dehydratase.

1. Aminolevulinate dehydratase (ALA-D) was studied in crude extract from mouse mammary carcinoma, normal mouse liver and tumour bearing mouse liver. 2. A Michaelis-Menten behaviour and Km values between 0.24 and 0.31 mM were obtained for the enzyme in either source. 3. In all three tissues there was a linear relationship between porphobilinogen formation and incubation time, up to 120 min, ALA-D was thermostable and optimum pH was at 6.8. 4. There seems to be no structural alterations in tumoural ALA-D as compared with the enzyme from liver of both normal and tumour bearing mice.

Animals↗
PubMed1990

An experimental assessment of cyanide challenge dose-response, time-course and recovery of indicative toxicity parameters.

1. Several sub-lethal doses of cyanide were assayed with the aim of obtaining significant differences in the parameters studied. A dose of 4 mg/kg s.c. was selected. 2. Present studies were carried out to determine the time dependence of the effects produced by s.c. administration of a single dose of potassium cyanide as well as the recovery time of some of the toxicity indicative parameters. 3. It was found that cyanide effects continued for at least 3 days; after the parameters had recovered normal values.

Animals↗
PubMed1990

In vivo prevention and reversal by the antimitotic colchicine and other related compounds of some porphyrinogenic drug action on heme pathway.

1. The effect of colchicine, vincristine and griseofulvin (GRIS) on the porphyrinogenic action of 2-allyl-2-isopropylacetamide (AIA) and veronal was studied in vivo and using the in vitro experimental model of tissue explant cultures. 2. Complete prevention by colchicine was found in liver and heart explant from animals treated with AIA and veronal. 3. Vincristine, GRIS and colchicine reversed AIA induction in liver explants, however reversal was partial or nil in skin and heart explants depending on the antimitotic and the tissue. 4. The usefulness of the combination of the in vivo experimental model and the in vitro explant tissue culture model, for this kind of studies is emphasized.

5-Aminolevulinate Synthetase↗
PubMed1990

The effect of cyanide intoxication on hepatic rhodanese kinetics.

1. Results of studies on the kinetics of hepatic rhodanese and the effects of S-adenosyl-L-methionine (SAM) on these kinetic parameters in cyanide-treated and non-treated mice are reported here. 2. The enzyme exhibited typical Michaelis-Menten behaviour with cyanide inhibition at concentrations higher than 50 mM. Km values of 4.74 and 0.85 mM were obtained for thiosulphate and cyanide, respectively, in control mice. 3. These results stress the biological importance of the rhodanese reaction for cyanide detoxification. 4. Km values were not significantly modified when the animals were intoxicated with a lethal (20 mg/kg) or a non-lethal (4 mg/kg) dose of cyanide. 5. SAM treatment either in control or in cyanide-poisoned animals doubled the Km's for cyanide.

Animals↗
PubMed1990

Cyanide intoxication. II. The effects of systematic cyanide challenge on indicative toxicity parameters.

1. Present studies were carried out to establish the action of cyanide maintained at a high level during a period as long as desired. 2. One of the earliest effects of cyanide seems to be the inhibition of hepatic rhodanese. These changes do not seem to occur in the blood. 3. Available cyanide labile-sulfur was always altered, unless cyanide tissue levels could not be detected. 4. The inhibitory action of cyanide on enzymatic reaction involving Schiff base intermediates was also corroborated through its effect on delta-aminolevulinate acid dehydratase activity. 5. S-Adenosyl-L-methionine was not able to modify the toxic cyanide action.

Animals↗
PubMed1990

Further evidence for an essential histidyl residue at the active site of pig liver 5-aminolevulinic acid dehydratase.

Photoxidation with methylene blue and rose bengal and chemical modification by diethylpryrocarbonate of pig liver 5-aminolevulinic acid dehydratase produced strong inactivation of the enzyme which was concentration dependent. Loss of enzyme activity by both photoxidation and ethoxyformylation was pH and time-dependent and protected by the presence of the substate and competitive inhibitors. The rate of inactivation was directly related to the state of protonation of histidyl groups, the unprotonated from being modified at a much faster rate than the protonated form. Plots of the pseudo-first order rate constants for 5-aminolevulinic acid dehydratase inactivation against pH resulted in typical titration curves showing inflection points at about pH 6.4 for methylene blue and rose bengal and 6.8 for diethylprocarbonate providing further and unequivocal evidence for the existence of critical histidyl groups at the active centre of the enzyme.

Animals↗
PubMed1990

Photodynamic and non-photodynamic action of several porphyrins on the activity of some heme-enzymes.

The action of porphyrins, uroporphyrin I and III (URO I and URO III), pentacarboxylic porphyrin I (PENTA I), coproporphyrin I and III (COPRO I and COPRO III), protoporphyrin IX (PROTO IX) and mesoporphyrin (MESO), on the activity of human erythrocytes delta-aminolevulinic acid dehydratase, porphobilinogenase, deaminase and uroporphyrinogen decarboxylase in the dark and under UV light was investigated. Both photoinactivation and light-independent inactivation was found in all four enzymes using URO I as sensitizer. URO III had a similar action as URO I on porphobilinogenase and deaminase and PROTO IX exerted equal effect as URO I on delta-aminolevulinic acid dehydratase and uroporphyrinogen decarboxylase. Photodynamic efficiency of the porphyrins was dependent on their molecular structure. Selective photodecomposition of enzymes by URO I, greater specificity of tumor uptake by URO I and enhanced porphyrin synthesis by tumors from delta-aminolevulic acid, with predominant formation of URO I, underline the possibility of using URO I in detection of malignant cells and photodynamic therapy.

Ammonia-Lyases↗
PubMed1990

Immobilized uroporphyrinogen I synthetase from Rhodopseudomonas palustris.

Rhodopseudomonas palustris uroporphyrinogen I synthetase (URO-S) has been chemically attached to Sepharose 4B and some of its properties have been studied. When 7-8 mg protein/ml activated Sepharose was used, immobilized URO-S retained 45% of the activity of the original soluble preparation, with a coupling yield of 66% after a period of 15 h. Optimal incubation conditions for the activity of gel-enzyme were determined. Unlike the soluble enzyme, the Sepharose-bound URO-S showed a biphasic substrate saturation curve, indicating that a protein conformational change had occurred during the process of immobilization. Immobilized URO-S stored at 4 degrees C for 35 days retained 90% of activity and when repeatedly used, up to 5 times, retained 48% of the original activity. Attachment of URO-S to Sepharose led to an enhanced thermal stability.

Ammonia-Lyases↗
PubMed1989

Glutamate:4,5-dioxovaleric acid transaminase from Euglena gracilis. Kinetic studies.

The kinetic properties of the enzyme L-glutamate:4,5-dioxovaleric acid aminotransferase (Glu:DOVA transaminase) from Euglena gracilis have been studied. 5-Aminolevulinic acid formation was linear with time for at least 45 min at 37 degrees C and L-glutamate was the most effective amino-group donor. Lineweaver-Burk double-reciprocal plots suggested a ping-pong reaction mechanism, with Km values for L-glutamate and DOVA of 1.92 mM and 0.48 mM respectively. Competitive parabolic substrate inhibition by DOVA at concentrations greater than 3.5-4.5 mM was observed. Glyoxylate (4-10 mM) was found to be a competitive inhibitor with respect to DOVA, whereas at low concentrations (0-4 mM) noncompetitive plots were obtained. An analysis of the possible enzyme forms involved, was carried out. In more crude preparations most of the enzyme is found to be in the form of an enzyme-glutamate complex.

Animals↗