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Biomedical subjects

A Yaron

Publications and source records attributed to A Yaron.

At least 55 records · Page 3Linked to original sources
PubMed1982

Absorption and distribution of orally administered jojoba wax in mice.

The liquid wax obtained from the seeds of the arid-land shrub jojoba (Simmondsia chinensis) is finding increasing use in skin treatment preparations. The fate of this wax upon reaching the digestive tract was studied. 14C-Labeled wax was administered intragastrically to mice, and the distribution of the label in the body was determined as a function of time. Most of the wax was excreted, but a small amount was absorbed, as was indicated by the distribution of label in the internal organs and the epididymal fat. The label was incorporated into the body lipids and was found to diminish with time.

Absorption↗
PubMed1982

Fluorogenic substrates for bacterial aminopeptidase P and its analogs detected in human serum and calf lung.

A sensitive fluorimetric assay was developed for bacterial aminopeptidase P, based on intramolecularly quenched fluorogenic substrates. Two substrates were synthesized. Phe(NO2)-Pro-HN-CH2-CH2-NH-ABz (substrate I) and Phe(NO2)-Pro-Pro-HN-CH2-CH2-NH-ABz (substrate II), in which the Phe(NO2) group (rho-nitro-L-phenylalanyl) quenches the fluorescence of the ABz group (omicron-aminobenzoyl). Both substrates were readily cleaved by aminopeptidase P from Escherichia coli, releasing rho-nitro-L-phenylalanine and causing a proportional increase in fluorescence. Complete hydrolysis of the two substrates resulted in a 7.5-fold and 3.4-fold fluorescence increase, respectively. Applying this fluorogenic assay, we were able to detect and measure quantitatively amino-peptidase P-like activity in the human serum and calf-lung extracts. Substrate II was shown to be specifically cleaved by aminopeptidase P in these preparations, while substrate I was apparently cleaved by other enzymes as well. In both preparations, the enzyme activity was independent of Co2+ ions, and Pro-HN-CH2-CH2-NH-ABz (Cbz) was inhibitory. The kinetic constant Km was determined as 0.35 mM and 0.28 mM for the human serum and the calf-lung enzymes respectively. The enzyme activity was only slightly dependent on pH in the range 7.0-8.4.

Aminopeptidases↗
PubMed1982

Angiotensin-I converting enzyme activity in the sera of captopril-treated hypertensive patients.

The blood pressure and serum angiotensin-I converting enzyme (SACE) activity were measured in captopril-treated hypertensive patients at frequent intervals. Inhibition of SACE was observed in patients responding to the treatment with lowering of blood pressure as well as in nonresponding cases. It was therefore concluded that a mechanism not depending on the formation of angiotensin-II by ACE is responsible for the high blood pressure persisting in the captopril treated nonresponding hypertensive patients. SACE activity in sera of captopril-treated patients recovers from inhibition when stored in frozen state at -20 degrees c. An apparent dissociation was therefore observed when regeneration of SACE activity during storage of sera was not taken into account. Meaningful activities are therefore obtained only if the assay is performed without prolonged storage.

Adult↗
PubMed1982

Captopril, an orally active angiotensin I converting enzyme inhibitor in the treatment of renovascular and essential hypertension.

The hypotensive response to captopril is described for 12 hypertensive patients, 7 of whom had renovascular hypertension. The drug was effective in lowering blood pressure. The few reversible adverse reactions that occurred included loss of the sense of taste in one patient and rash and fever in another. Three patients with renal failure showed deterioration of renal function during treatment, suggesting the advisability of treating such cases with lower dosages.

Adult↗
PubMed1979

A fluorimetric assay for angiotensin-I converting enzyme in human serum.

A fluorimetric method is described for a simple, sensitive and reproducible assay for angiotensin-I converting enzyme in human and guinea pig sera. The very weak fluorescent substrate o-aminobenzoylglycyl-p-nitro-L-phenylalanyl-L-proline is enzymatically hydrolyzed, producing the highly fluorescent o-aminobenzoylglycine that is quantitatively determined by spectrofluorimetry. Dependence of activity on substrate concentration, amount of serum, time of incubation and pH were investigated. The KM value for the substrate is 0.1 and 0.032 mM for the human and guinea pig serum enzyme, respectively. The mean value of serum angiotensin-I converting enzyme for 16 normal adult persons was 2.56 +/- 0.10 (S.E.) with a standard deviation of 0.81 nmol/min/ml serum.

Animals↗
PubMed1978

An intramolecularly quenched fluorescent tripeptide as a fluorogenic substrate of angiotensin-I-converting enzyme and of bacterial dipeptidyl carboxypeptidase.

The N-acyltripeptide 2-aminobenzoylglycyl-p-nitrophenylalanylproline was synthesized and applied as a substrate in the assay of angiotensin-I-converting enzyme from calf lung and human serum, and of the bacterial dipeptidyl carboxypeptidase from Escherichia coli. This compound belongs to a new class of substrates for proteolytic enzymes, having the general structure F--X--Q in which fluorescence of group F is quenched by intramolecular interaction with the group Q. Enzymatic cleavage of the peptide chain (X stands for one or more amino acid residues) generates the unquenched F-containing derivative and the resulting fluorescence is used for quantitative measurement of the hydrolysis rate. Cleavage of the Gly-Phe(NO2) peptide bond in the weakly fluorescent 2-amino-benzoylglycyl-p-nitrophenylalanylproline molecule results in appearance of the 71 times higher fluorescence (lambdamax = 415 nm) of 2-aminobenzoylglycine. Continuous recording of the rising fluorescence allows convenient, sensitive and specific determination of the enzymatic activity, applicable to crude enzyme preparations and human serum. The activity of the mammalian enzyme, measured by this method, is enhanced by Cl- ions and inhibited by low concentrations of EDTA and [Asn1, Val5]angiotensin II. Kinetic measurements showed Michaelis-Menten behavior, Km = 0.21 +/- 0.1 mM and 0.16 +/- 0.1 mM for the calf lung and the bacterial enzyme respectively.

Carboxypeptidases↗
PubMed1978

Specificity and genetic restrictions of the guinea-pig immune response to dinitrophenyl-lysyl-alanyl octapeptides.

A series of 2,4-dinitrophenyl (DNP) octapeptides containing L-lysine and L-alanine were prepared to examine the specificity and genetic restrictions of both cellular and humoral immune responses in inbred guinea-pigs. Strains 2 and 13 guinea-pigs were therefore immunized with Lys4-Ala3-Lys(DNP), Lys3Ala4-Lys(DNP), Lys2-Ala5-Lys(DNP) and Lys-Ala6-Lys(DNP). Only Lys4-Ala3-Lys(DNP) was under Ir gene control and could induce both antibody and T-cell responses in strain 2 guinea-pigs. In contrast, Lys4-Ala3-Lys(DNP) injected in strain 13 guinea-pigs and the other DNP-octapeptides injected in strain 2 or 13 guinea-pigs elicited only antibody formation and no specific T-cell mediated response. Antibody formed in the absence of specific T-cell responsiveness in either strain 2 or 13 was hapten specific and lacked the capacity to discriminate the immunizing antigen from closely related DNP-peptides. Antibody produced by animals with specific T-cell responses, on the other hand, was exquisitely specific for the immunizing peptide and could discriminate it from closely related peptides.

Alanine↗
PubMed1977

Intramolecularly-quenched fluorescent peptides as fluorogenic substrates ofleucine aminopeptidase and inhibitors of clostridial aminopeptidase.

Fluorogenic oligopeptide derivatives of the type Lys(ABz)-ONBzl, where ABz iso-aminobenzoyl (anthraniloyl), X stands for Ala Phe, or Ala-Ala, and ONBzlis p-nitrobenzyloxy, were synthesized and shown to be hydrolyzed by leucine aminopeptidase. The hydrolysis is accompanied by an increase in fluorescence due to disruptionof the intramolecular quenching of the fluorescent anthraniloyl moiety by the nitrobenzyester group. The spectral characteristics of the compounds are not consistent withan energy transfer mechanism according to Förster, therefore the quenching isassumed to be caused by a direct encouter between the quenching and the fluorecentgroups. The change in fluorescence that accompanies the enzymic hydrolysis ofthe first peptide bound was used for quantitative measurement of the activity ofthe activity of leucine aminopeptidase and for the determination of some of itskinetic parameters. A bacterial aminopeptidase from Clostrdium histolyticumthat is very similar to leucine aminopeptidase in its substrate specificity inits substrate specificity did not hydrolyze the above peptidederivatives. Thehydrolysis of leucine p-nitroanilide by this enzyme was found to be inhibitedby the three peptides and the corresponding inhibition constants were determined.

Aminopeptidases↗
PubMed1977

Immune ascites in the guinea pig: specificity of cells and antibody in an induced peritoneal exudate.

Strain 2 guinea pigs, immunized with Lys10-Lys(Dnp) in CFA and repeatedly injected intraperitoneally with adjuvant, developed ascites. The fluid was harvested over 8 months in total amounts up to 2 liters per animal and contained substantial amounts of cells and antibody which reacted with the immunizing antigen and related peptides. The antibody was of the IgG and IgA classes and showed restricted heterogeneity. Among synthetic Dnp-oligopeptides, both the cells, studied by antigen-stimulated thymidine incorporation, and the purified antibody, studied by fluorescence quenching, demonstrated the same specificity for the immunizing antigen as has previously been noted in lymph node cells and in serum antibody. The technique offers a means for studying more cells and more antibody than has previously been possible from individual guinea pigs.

Animals↗
PubMed1976

An extracellular aminopeptidase from Clostridium histolyticum.

An aminopeptidase was isolated from the culture filtrate of Clostridium histolyticum and purified to homogeneity. Absence of endopeptidase activity in the purified preparation was demonstrated. Gel filtration on a calibrated column indicates an apparent molecular weight of 340000 for the native enzyme. Gel electrophoresis of the denatured enzyme in the presence of dodecylsulfate in constant acrylamide concentration and in a concentration gradient, resulted in the appearance of a single component for which a molecular weight of 51000 and 59000 respectively, was calculated. From mobilities of crosslinked and denatured protein species a molecular weight of 56000 was obtained for the monomer. Specificity studies show that the enzyme cleaves all types of N-terminel amino acid residues including proline and hydroxyproline from small peptides and from polypeptides. The peptide bond formed between an N-terminal amino acid residue and proline is not cleaved by the enzyme. The combined action of aminopeptidase-P and clostridal aminopeptidase leads to complete hydrolysis of the proline-rich nonapeptide bradykinin. Low rates of hydrolysis was observed for charged residues, and amides of amino acids. Kinetic studies with five tripeptides of the general structure X-Gly-Gly, where X stands for Leu, Phe, Val, Ala, or Pro, show a decrease in Km with the increasing size of the hydrophobic side chain of X. The highest Kcat values are observed with proline and alanine. In the series Pro-Gly, Pro-Gly-Pro, Pro-Gly-Pro-Pro, the last peptide is the best substrate, indicating an active site complementary to at least four amino acid residues. The enzymatic activity is dependent on the presence of divalent cations, maximal activation being reached with Mn2+ and Co2+. The optimal pH for the Mn2+ and Co2+- activated enzyme is 8.6 and 8.2 respectively. The optimal temperature is 40 degrees C. Inhibition of the aminopeptidase was achieved with Zn2+, Cu2+ and p-mercuribenzoate, but not with diisopropylphosphofluoridate.

Amino Acids↗