Reflections on the university/industry partnership.
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Biomedical subjects
Publications and source records attributed to M Anbar.
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Using thin-layer chromatography fluorometry, gas chromatography mass spectrometry and field ionization mass spectrometry the findings of Green et al. regarding the identification of 5-methoxytryptamine in rat hypothalamus could not be confirmed. On derivatization using pentafluoropropionic anhydride, melatonin underwent partial transacylation to yield 5-methoxytryptamine as one of the by-products. We thus consider the finding of 5-methoxytryptamine in rat hypothalamus by mass fragmentography as an artifact forming from melatonin, which is normally present in rat hypothalamus.
Metabolic profiles of urine extracts of humans with viral infections, as well as of media of virus-infected human tissue cultures, have been analyzed by non-fragmenting mass spectrometry and compared with corresponding controls. The spectra were then subjected to several alternative computerized statistical procedures to detect diagnostic biochemical profiles. Controlled longitudinal studies on fully informed, consenting volunteers who received sandfly fever virus demonstrate the onset of a characteristic metabolic pattern that precedes the onset of symptoms and subsides when the patients overcome the infection. Longitudinal studies of human tissue cultures infected with poliomyelitis virus demonstrate characteristic metabolic patterns within a few hours after infection. Non-fragmenting mass spectrometry may thus provide the clinical laboratory with a sensitive, reliable test for viral infections significantly faster than attainable by current techniques.
A new methodology for comparative bioavailability testing is described in which each drug formulation is compared with a stable isotope-labeled variant of the drug that is consumed orally in solution at the same time the tested formulation is ingested. The methodology is used to determine the comparative bioavailabilities of two commercially available brands of imipramine hydrochloride. The power of the new methodology to detect differences between drug formulations, when, in fact, such differences exist, is shown to be superior to that of conventional bioavailability tests.
Fluoride release from fluorine-carrying copolymers of vinylphosphonate induced by calcium apatite and tooth enamel has been investigated. Fluoride ions were determined potentiometrically in the study of calcium hydroxyapatite, and Auger spectroscopy was used to study the fluoride release to enamel. The adsorbed copolymer of vinylphosphonic acid and vinylphosphonyl thiofluoride was shown to release fluoride to calcium hydroxyapatite and to enamel. The oxygen analog has shown a similar behavior with calcium hydroxyapatite, but not with tooth enamel. The results suggest a potential application of such copolymers as caries preventive agents which combine the effects of polyphosphonates and of fluoride ions.
A quantitative method is reported for the determination of imipramine in plasma samples in the low nanogram and subnanogram range. The sensitivity and precision of the technique, which involves high pressure liquid chromatography and direct probe field ionization mass spectrometry, are approximately an order of magnitude greater than are offered by gas chromatography mass spectrometry with selected ion monitoring using deuterated or other types of internal standards. [2H6]Imipramine, labeled in the ethylene bridge and in the aromatic rings, serves as the isotopic diluent. The method has been used for the determination of the comparative bioavailabilities of two different commercial preparations of imipramine. In these tests, subjects ingested a 25 mg tablet of one or the other drug preparation together with a solution containing an equivalent amount of imipramine deuterated in the ethylene bridge ([2H2]imipramine). The latter served as an internal check for intrasubject variability in absorption of the imipramine tablets. Typical results from one of the subjects are presented.
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Multilabelled, non-radioactive thymidine and tritiated thymidine were incorporated simultaneously into the DNA of a rapidly-growing, transplantable mouse lymphoma and the DNA of mouse small intestine by a single intraperitoneal injection of both labelled nucleosides. Mice were killed at selected times during the next 7 days, and the specific activities of the tissues and extracted DNA and the 132/126 mass ratio of thymine isolated from the DNA were determined by scintillation counting and by field ionization mass spectrometry respectively. The rates of decrease of the concentrations of tritiated and stable isotope-labelled thymine in the DNA of the tumour or of the intestine were essentially identical. These results indicate the feasibility of using thymidine multilabelled with stable isotopes for measurement of cell turnover rates in conjunction with cancer therapy.
The thin-layer chromatography of imipramine on silica gel plates was studied in fifteen solvent systems. The mobility of imipramine labeled with deuterium in the methyl groups of the dimethylaminopropyl side chain differs markedly from that of unlabeled imipramine. Partial or complete separations between unlabeled and deuterated imipramine were observed in all basic and neutral solvent systems investigated, but not in weakly acidic solvents. Isotopic fractionations of imipramine were also found on alumina thin-layer plates, but were not detected in cellulose chromatography. In all thin-layer isotopic separations, the unlabeled compound migrates more rapidly than the deuterated molecule. These results can be explained by a stronger basicity of deuterated imipramine relative to its unlabeled counterpart.
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A method has been developed for the measurement of DNA synthesis in vivo using the incorporation of multilabeled, non-radioactive thymidine. Simultaneous intraperitoneal injection of hexalabeled thymidine and tritiated thymidine into a normal adult rat resulted in the incorporation of both labeled nucelosides into the DNA of cells undergoing replication. The DNA of several tissues and organs was analysed, including liver, thymus, spleen, bone marrow, and small intestine. Following extraction with hot trichloroacetic acid, acid hydrolysis, and thin-layer chromatography of the hydrolysates, the isotopic compositions of the thymine products were determined by field ionization mass spectrometry and by scintillation counting. The relative incorporation of radioactive and stable isotope-labeled thymidine was similar in all tissues, and corresponded to the ratio of the two labeled nucleosides in the injected material. These results indicate the feasibility of utilizing thymidine multilabeled with stable isotopes for measurement of cellular proliferation rates in conjunction with cancer therapy.
The adhesion of acrylic composite restorative materials to etched dental enamel is increased by a factor of two or more when the enamel is coated with a thin layer of vinyl-benzyl phosponic acid (VBPA) prior to forming the enamel-restorative bond. The present results support the suggestion that the enhanced adhesion is attributable to bonding between calcium ions in the enamel surface and phosphonate groups in the VBPA.
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Field ionization mass spectrometry has been applied to multicomponent analysis of metabolites in human urine for the diagnosis of metabolic disorders, exemplified here by infectious hepatitis. The molecular weight profiles of carboxylic acids and of "neutral" metabolites in the urine of patients with infectious hepatitis were compared with those in urine of normals. The "neutral" metabolites showed 24 characteristic spectral differences in the range 68 to 215 atomic mass units, which provided correct diagnoses in 100% of the cases and a "diagnostic power" of unity. These results are even more encouraging than those obtained earlier with the acidic metabolites, where 11 mass numbers were found in the same mass range to be useful for correct diagnoses in 91% of the cases. The measurements were performed on urine samples preextracted on Amberlite XAD-2 columns. We used a quadrupole mass spectrometer interfaced with a multichannel analyzer for mass analyses.
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