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Biomedical subjects

D Bartos

Publications and source records attributed to D Bartos.

At least 37 records · Page 2Linked to original sources

Polyamines and polyamine complexes: analytical comments.

Different methods of sample preparation for determination of di- and polyamines (putrescine, spermidine, spermine) and their conjugates in serum, plasma, and urine are discussed. Higher polyamine values obtained after hydrolysis of serum or plasma acidic extracts suggest the presence of polyamine conjugates. Additional polyamines might be bound to proteins and released by hydrolysis of the crude samples. Biological studies of bound polyamines in health and disease will require procedures for quantitative analysis. For the pertinent measurements, radioimmunoassays employing antibodies developed against isolated polyamine conjugates may prove useful.

Acetylation↗

Development of a specific antispermine antibody.

Highly specific antibody to spermine was produced in the rabbits by immunization with a spermine-thyroglobulin conjugate of high molar ratio (740:1). Cross-reactivity with spermidine was 1.4-2.2 percent and negligible with putrescine and other compounds of similar structure. The average affinity constant of five pools of antispermine rabbit antisera was Ka = 1.8 +/- 0.2 X 10(8) liter/mole.

Animals↗

Polyamines: an unrecognised cardiovascular risk factor in chronic dialysis?

The significance of raised polyamine (P.A.) levels in chronic-dialysis patients is unknown. Since these biologically active substances have hormone-like properties and promote cell-growth in plant and animal tissues, it is possible that they stimulate proliferation of arterial smooth-muscle cells (S.M.C.)--a central process in atherogenesis--and thereby contribute to the rapidly accelerated cardiovascular disease observed during dialysis. Such a role for P.A. is supported by tissue-culture studies, which show not only that P.A.-rich serum from dialysis patients stimulates S.M.C. growth, but also that this mitogenic effect is lost when P.A. are selectively removed from uraemic serum and restored by their addition. Although these observations provide new insights into possible mechanisms of atherogenesis, they are not surprising in view of the many known biological actions of P.A.

Arteries↗

Purification of rabbit antispermine antiserum by affinity chromatography.

Immunoglobulins G were isolated from a crude rabbit antispermine antiserum using a protein A-Sepharose CL-4B column. Affinity chromatography coupling spermine to the Sepharose matrix was employed for separation of antispermine antibodies from these immunoglobulins. Four fractions of antispermine antibodies were isolated by stepwise elution with solutions at pH range from 4 to 1. Binding constants were determined and cross-reactivity with other polyamines was tested for each fraction of antibodies. Highly specific antispermine antibodies were found in fraction IV.

Animals↗

1-N-Acetylspermidine: occurrence in normal human serum.

Using gas chromatography mass spectrometry following derivatization (N-trifluoroacetylation), 1-N-acetylspermidine was found to occur in normal human serum at a level of approximately 0.008--0.5 nmol ml(-1), i.e. about two orders of magnitude lower than the corresponding spermidine level. No evidence for the occurrence in serum of the isomeric 8-N-acetylspermidine was found. Mass spectra of authentic samples of trifluoroactylated 1-N- and 8-N-acetylspermidine are presented.

Acetylation↗

Radioimmunoassay of spermidine in human serum.

Specific antispermidine antibodies were produced following immunization of New Zealand white rabbits with a spermidine-thyroglobulin conjugate of high (greater than 200:1) hapten-carrier molar ratio. The harvested antispermidine rabbit antiserum was characterized and a radioimmunoassay procedure developed. The specificity of this antibody was tested against cadaverine, putrescine, spermine, monoacetylputrescine, N-8-acetylspermidine and other compounds of similar molecular structure. Negligible cross-reactivity was observed with putrescine (2.1%), cadaverine (0.6%), spermine (0.3%), diaminopropane (less than 0.1%), and no cross-reaction was found with monoacetylputrescine, N-8-acetylspermidine, histamine, L-lysine and L-ornithine. The spermidine values on serum of five healthy volunteers were determined by RIA and compared with values by gas chromatographymass spectrometry. Statistical evaluation of the results were highly agreeable. Antibody titer, intra-assay precision and analytical accuracy were tested. The assay requires only 20 microliter of serum and has picomole sensitivity. The sensitivity, specificity and small sample requirement make this procedure an effective tool for spermidine analysis.

Animals↗

Stereospecific antibodies to methadone. II. Synthesis of d- and 1-methadone antigens.

Diastereomeric methadols were prepared by reduction of the methadone enantiomers. Resulting methadols, after purification and characterization by mass spectrometry, were succinylated. The hemisuccinyl derivatives were conjugated with thyroglobulin via carbodiimide condensation. The conjugates of d- and of 1-methadol hemisuccinyl-thyroglobulin were employed as the antigens for immunization of white New Zealand rabbits in order to obtain specific anti d- and anti 1-methadone antisera.

Animals↗

Stereospecific antibodies to methadone. I. Radioimmunoassay of d,l-methadone in human serum.

Anti-d,l-methadone antibodies were produced in rabbits immunized with d,l-methadol-hemisuccinate thyroglobulin conjugate. Using the antiserum, a radioimmunoasay (RIA) for determination of d,l-methadone in human serum has been developed and is described. Concentration of d,l-methadone of 1.4 pmol in a native serum sample (volume 0.1 ml or less) could be measured directly by RIA. The antibodies crossreact 100% with d,l-methadone, 50% with d-methadone, 50% with l-methadone and 100% with alpha-d-methadol. No crossreactivity was found with alpha 1-methadol, morphine, meperidine, dextropropoxyphene, 2-ethyl-5-methyl-3,3-diphenyl-l-pyrroline and 2-ethylidene-l, 5-dimethyl-3,3-diphenylpyrrolidene. High sensitivity and small sample requirements make this method suitable for future monitoring of patients on methadone maintenance and for studies where other procedures have lack of sensitivity.

Animals↗

Normal functions of the thyroid gland of the pygmy goat.

Normal values were obtained for blood serum PBI, T3 index. T4, and cholesterol in a colony of pygmy goats (n equal to 55) of mixed sex and age. Serum PBI values averaged 8.1 plus or minus 1.2 mug/dl with no significant sex differences. The mean T3 index and T4 value were 1.1 plus or minus 0.1 and 7.2 plus or minus 1.1 mug/dl, respectively, with no sex differences. The mean serum cholesterol value was 90.0 mg/dl, with sex differences apparent. Serum cholesterol averaged 84.9 mg/dl (n equal to 44) for females and was significantly higher in intact males (97.4 mg/dl; n=8) and significantly lower in castrate males 69.2; n=6. There was a consistent and significant increase in cholesterol values with age in females, an unexplained phenomenon also observed in humans. There was no evidence of thyroid malfunction in the animals studied.

Age Factors↗

Direct determination of polyamines in human serum by radioimmunoassay.

Antipolyamine antibodies were produced following immunization of New Zealand white rabbits with thyroglobulin spermine conjugate, using multiple-site injection and macrophage-harvesting techniques. The antispermine antibody in a radioimmunoassay system cross-reacted with spermidine (22%), putrescine (1%), diaminopropane (6%), and cadaverine (0.16%). No reaction with L-lysine, ornithine, or histamine was noted. Using this antibody, the radioimmunoassay for direct determination of serum polyamines was developed. A sensitivity of 200 pg was achieved on human serum samples. This radioimmunoassay would appear to be a useful tool for the detection and monitoring of a variety of pathophysiological states associated with abnormal polyamine metabolism, i.e., cancer.

Animals↗