Biomedical subjects
Publications and source records attributed to I Mayer.
Carbonated hydroxyapatites in the presence of F- and Li+ were synthesized. Limited uptake (0.2-1.6 ppm) of Li+ by the apatites was observed, which was proportional to the initially added Li+ and the carbonate concentration of the samples. Fluoride has retarding effect on the Li+ uptake. Li+ causes a decrease in the solubility of carbonated hydroxyapatite.
The binding of the 3'-[1,5-(dimethylamino)naphthoyl] (DAN) derivatives of AMP, ADP, and ATP to the solubilized ADP/ATP carrier is studied, evaluating primarily the fluorescence enhancement and 3H-labeled compound binding. DAN nucleotides also fluoresce when adsorbed to Triton X-100 micelles that are used for solubilization of the carrier. The partition of DAN-AMP between water and Triton X-100 micelles is measured, and it is shown to be shifted toward a higher content in Triton micelles with increasing salt concentration. In order to maintain a low level of fluorescence, the Triton content is decreased. The fraction of DAN nucleotide fluorescence due to carrier binding is determined by the suppression with bongkrekate (BKA). In contrast to the membrane-bound carrier, the solubilized preparation shows an increase of total BKA-sensitive fluorescence by 30-60% upon addition of ATP or ADP. In the solubilized atractylate-protein complex, the ADP-stimulated fluorescence amounts even to 80%. The suppression of fluorescence by BKA is independent of the presence of ADP or ATP, while that by carboxyatractylate (CAT) depends on ADP or ATP. The quantitation with [3H]BKA and [3H]CAT of these ligand interactions with DAN-AMP fluorescence shows that DAN-AMP fluorescence reflects the "m"-state carrier population and its redistribution under the influence of ADP or ATP. Thus, besides the "c"/"m" distribution, the kinetics of the c to m transition in the solubilized carrier also can be determined. The m share is increased to 80% when SO4, Pi, or pyrophosphate is present during solubilization. The rate of the ADP- or ATP-stimulated transition to the m state is markedly dependent on pH and on the presence of various anions, whereas the extent is little varied. The affinity decreases 4-fold going from DAN-AMP to DAN-ADP and to DAN-ATP (KD = 0.9, 1.6, and 3.2 microM). Comparison with physical binding of [3H]DAN nucleotides shows that the fluorescence yield of bound DAN-AMP is about 1.4 times higher than that of bound DAN-ATP. DAN substitution causes more than a 100-fold affinity increase for AMP and a 50-fold increase for ADP or ATP, probably because of interaction of the DAN group with a hydrophobic niche. A less specific, low-affinity displacement of DAN nucleotides by GDP, ADP, GTP and ATP (Ki = 1-2 mM) probably reflects primarily the ionic interactions at the binding center.
Iron uptake (Fe2+ and Fe3+) by bones, teeth, and dental enamel was studied, in vivo and in vitro, by chemical, powder X-ray diffraction and Mossbauer spectroscopy methods. Atomic absorption tests have revealed the permanent uptake of small amounts of iron by dental enamel soaked in vitro in solutions containing Fe2+. Mossbauer spectra show that the iron attached to the dental enamel stays at the same valency it had in the soaking solutions. Mossbauer measurements of in vivo samples show that iron is present in bones and teeth mainly as Fe3+ (10% Fe2+ in teeth), in compound similar to FeOOH. Iron is released or exchanged from teeth at a much lower rate than from bones.
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Human recombinant gamma-interferon (rhu-IFN-gamma) and human recombinant alpha-interferon (rhu-IFN-alpha 2 arg) with a chemical purity of over 95% were compared for their antiproliferative and HLA-DR-inducing activity in five human breast cancer cell lines (BT 20, ZR 75.1, MCF 7, 734B, Hs578T). Cytostatic effects on tumor cells were evaluated in monolayer cultures. HLA-DR antigen expression was examined by an indirect immunofluorescence technique using two different anti-HLA-DR monoclonal antibodies (anti-HLA-DR, VID-1) against framework determinants. rhu-IFN-gamma and rhu-IFN-alpha 2 arg differed in their antiproliferative efficiency in terms of both dose dependency and the spectrum of sensitive target cells. Combinations of rhu-IFN-gamma and rhu-IFN-alpha 2 always resulted in higher cytostatic effects. HLA-DR expression was exclusively inducible by rhu-IFN-gamma and did not correspond to its antiproliferative activity. Furthermore, HLA-DR expression did not depend on proliferation but did require intact RNA and protein syntheses as shown by inhibition with cycloheximide and actinomycin D. HLA-DR antigen expression in mammary cancer lines was dependent on time, dose, and the continued presence of rhu-IFN-gamma. Thus, our data suggest that in particular combinations type I and type II interferons might be useful in the treatment of breast cancer because they provide effective cytostatic and cell membrane-modulating properties.
Using established breast cancer cell lines in a cell culture model we studied the growth effect of retinoic acid (RA) alone or in combination with the antiestrogen 4-hydroxytamoxifen (OHT). Cytoplasmic 3HRA binding sites were determined by sucrose density gradient centrifugation analysis. Of the three cell lines Hs578T, BT 20, and 734 B only the last showed a significant amount of specific RA binding (10(5) sites/cell). This cell line showed a dose dependent decrease in proliferation after a long-term incubation with RA whereas the 3H-thymidine uptake was highly significantly increased after incubation with 10(-6)M of RA for 20 hr. Growth inhibition was not further increased by the addition of OHT (10(-6) M), but the increase in thymidine incorporation due to RA was neutralized by OHT. Hs578T and BT 20 cells were not affected by any of the treatments. The different action of RA on proliferation and thymidine incorporation suggests a cell cycle specific mechanism.
Fluorescent 3'-O-acyl-substituted adenine nucleotide (dimethylamino)naphthoyl and trinitrophenyl groups were studied for binding to the ADP/ATP carrier in mitochondria and submitochondrial particles. The changes in fluorescence intensity and emission maximum are for the most part similar to those observed in nonaqueous solvents. The (dimethylamino)naphthoyl derivatives from a largely quenched aqueous state have a shortwave shift up to 85 nm and increase up to 90-fold (1,5 derivative), whereas the little quenched naphthoyl derivatives show a fluorescence decrease and the weakly fluorescent trinitrophenyl derivative shows only a small increase on binding. All derivatives are good inhibitors (K1 = 1-10 microM) of nucleotide transport. The fluorescence titrations have an apparent K1/2 = 2-7 microM. The fluorescence of the 1,5-DAN nucleotide is fully suppressed by bongkrekate but only partially suppressed by carboxyatractylate. The fluorescence response is much stronger in submitochondrial particles than in mitochondria. Both facts suggest fluorescent binding to the "m" state of the carrier site at the inner face of the membrane. 1,5-DAN-AMP shows a slightly more efficient binding than DAN-ADP or DAN-ATP.
The binding to the ADP/ATP carrier in mitochondrial membranes of the 3'-O-(dimethylamino)naphthoyl (DAN) derivatives of AMP, ADP, and ATP was quantitatively analyzed. The sidedness of the fluorescent type binding to the "m" side only was shown comparing the mitochondrial membranes in various stages of integrity and surface orientation. In particles displacement by bongkrekate (BKA) is direct, whereas in the case of carboxyatractylate (CAT) the requirement for ADP and ATP demonstrates the transition from the "m" to the "c" state. Quantitatively the "physical" binding of [3H]DAN-AMP and fluorescence are well correlated, allowing for a little nonfluorescent binding to the c side. For DAN-AMP KD is 1.6 microM, for DAN-ADP KD is 0.8 microM, and in the Hill plot a straight line with n = 1.25 is obtained. The maximum number of binding sites for [3H]DAN-AMP (1.5 mumol/g of protein) is about equal to the sites found for [3H]BKA if the unspecific binding of both ligands is differentiated by blocking carrier sites with CAT. [3H]CAT binding is somewhat lower in accordance with the limited access of CAT to inverted vesicles. ADP is able to decrease fluorescence only by about 35% at high concentrations (10 mM) whereas GDP has virtually no effect. With ADP, DAN-AMP binding decreases by 30% of the total binding sensitive to BKA. Binding to ATPase is low because of the absence of Mg2+.(ABSTRACT TRUNCATED AT 250 WORDS)
Apatites containing Na, Mg, and CO3 in amounts occurring in tooth enamel mineral were synthesized by precipitation from aqueous solutions. The X-ray diffraction patterns showed only apatitic reflections, which were somewhat broader than those of enamel. X-ray Guinier photographs resulted in diffuse reflections from which the lattice parameters could not be determined. The infrared (IR) absorption spectrum of the synthetic samples was practically identical with that of tooth enamel. However, after heating the samples for 1/2 h at a temperature between 300 and 600 degrees C, no absorption peak was found at 2340 cm-1 as in the IR spectrum of tooth enamel after heating.
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Rats were given drinking water containing up to 300 parts/10(6) F and the conversion of hydroxyapatite to fluoridated apatite was estimated chemically and by X-ray diffraction. The differences of the mean fluoride uptakes from the very different concentrations of fluoride given were significant. Prolonged washing of the bones with demineralized water released 5-10 per cent from the fluoride uptake. Formation of fluoridated apatite was evident by decreases in the a parameter of the hydroxyapatite phase. In the washed bones, there was no appreciable transformations in the a parameters. The co-existence of Mg-containing beta-Ca3(PO4)2 with the apatite phase was detected in heated bone samples. The amount of beta-Ca3(PO4)2, its dependence on the age of the bones and its Mg content are interdependent.
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