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Biomedical subjects

B Senger

Publications and source records attributed to B Senger.

51 records · Page 3Linked to original sources

Human amelogenesis: high resolution electron microscopy of nanometer-sized particles.

Growth of inorganic crystals of enamel is described as a two-stage process with growth of ribbon-like crystals in length and width, followed by their development in thickness. In early stages of crystal growth during human amelogenesis nanometer-sized particles with a mean diameter of 1.1 nm were described between ribbon-like crystals. These small particles had a crystalline structure but their lattice parameters did not seem to be directly related to those of calcium phosphates. The nanometer-sized particles appear to correspond to initial stages of apatite crystal growth. Their localization close to ribbon-like crystals and their progressive increase in size and number may indicate that they represent a precursor phase for these crystals. Nucleation areas at both extremities, of elongated ribbon-like crystals could be involved in the two-directional growth of ribbons and/or in nanometer-sized particle nucleation.

Amelogenesis↗

Subsets of neurotensin-immunoreactive neurons in the rat striatal complex following antagonism of the dopamine D2 receptor: an immunohistochemical double-labeling study using antibodies against Fos.

A study was done to determine if the Fos and neurotensin immunoreactivities elicited in the rat striatal complex by the selective dopamine D2 receptor antagonist, S(-)-eticlopride hydrochloride are co-localized in the same neurons. Following injections of eticlopride, Fos and neurotensin immunoreactivity were both non-uniformly distributed among the striatal compartments and subterritories. Fos was co-localized in a significant number of small, lightly neurotensin-immunoreactive neurons, but not in a larger subset of neurons with significantly greater median diameter that exhibited intense neurotensin immunoreactivity extending well into the dendritic arbor. It is proposed that neurotensin-immunoreactive neurons lacking Fos immunoreactivity are prominent following selective blockade of the dopamine D2 receptor and represent a subset of striatal neurotensin-immunoreactive neurons. Neurotensin-immunoreactive cells containing Fos nuclei represent a distinct subset, possibly the one that is dominant following administration of reserpine [Zahm (1992) Neuroscience 46, 335-350]. Insofar as Fos expression has been reported to accompany activation of striatonigral and striatopallidal neurons, the absence of Fos in the subset of neurotensin neurons displayed following D2 receptor blockade may be at odds with activation and perhaps is more consistent with inactivation and accompanying decreased release of neurotensin [see Frey et al. (1988) Neurochem. Int. 12, 33-38, and Bean et al. (1989) J. Neurosci. 9, 4430-4438] as a mechanism underlying the accumulation of neurotensin in that subset of striatal neurons.

Animals↗

The anticodon triplet is not sufficient to confer methionine acceptance to a transfer RNA.

Previous work suggested that the presence of the anticodon CAU alone was enough to confer methionine acceptance to a tRNA. Conversions of Escherichia coli nonmethionine tRNAs to a methionine-accepting species were obtained by substitutions reconstructing the whole methionine anticodon loop together with preservation (or introduction) of the acceptor stem base A73. We show here that the CAU triplet alone is unable to confer methionine acceptance when transplanted into a yeast aspartic tRNA. Both non-anticodon bases of the anticodon loop of yeast tRNA(Met) and A73 are required in addition to CAU for methionine acceptance. The importance of these non-anticodon bases in other CAU-containing tRNA frameworks was also established. These specific non-anticodon base interactions make a substantial thermodynamic contribution to the methionine acceptance of a transfer RNA.

Anticodon↗

Configurations of adsorbed hard spheres after diffusion in a gravitational field.

The deposition and adhesion of particles on a solid surface are governed by a great number of interplaying forces. In this paper we analyze, by means of computer simulations, the influence of (i) the short-range repulsive forces, modeled by hard sphere interactions, (ii) the gravitational forces, and (iii) the diffusion process of the particles in the fluid on the structure of the surface covered by the particles. In particular, the evolution of the limiting coverage, [symbol, see text] infinity (where [symbol, see text] is the reduced relative surface coverage), and the radial distribution, g(r), at the jamming limit, are determined as a function of the gravitational forces. These forces play an important role in many experiments performed on latex beads. Our results should stimulate new experiments in this field and, thus, be directly experimentally tested. It is shown, for example, that for polystyrene particles [symbol, see text] infinity is constant and equal to the random sequential adsorption jamming limit value for radii R not larger than 1 micron. It increases for 1 </= R </= 3 micron and tends, for higher R, to a plateau, whose value is approximately equal to 0.61. The tendency to a closer packing when R is large, and thus large gravitational forces, is confirmed by the shape of the radial distribution function. This phenomenon occurs not only for jammed surfaces but also for unsaturated surfaces.

Journal Article↗

Binding of the yeast tRNA(Met) anticodon by the cognate methionyl-tRNA synthetase involves at least two independent peptide regions.

As for Escherichia coli methionine tRNAs, the anticodon triplet of yeast tRNA(Met) plays an important role in the recognition by the yeast methionyl-tRNA synthetase (MetRS), indicating that this determinant for methionine identity is conserved in yeast. Efficient aminoacylation of the E. coli tRNA(Met) transcript by the heterologous yeast methionine enzyme also suggests conservation of the protein determinants that interact with the CAU anticodon sequence. We have analysed by site-directed mutagenesis the peptide region 655 to 663 of the yeast MetRS that is equivalent to the anticodon binding region of the E. coli methionine enzyme. Only one change, converting Leu658 into Ala significantly reduced tRNA aminoacylation. Semi-conservative substitutions of L658 allow a correlation to be drawn between side-chain volume of the hydrophobic residue at this site and activity. The analysis of the L658A mutant shows that Km is mainly affected. This suggests that the peptide region 655 to 663 contributes partially to the binding of the anticodon, since separate mutational analysis of the anticodon bases shows that kcat is the most critical parameter in the recognition of tRNA(Met) by the yeast synthetase. We have analysed the role of peptide region (583-GNLVNR-588) that is spatially close to the region 655 to 663. Replacements of residues N584 and R588 reduces significantly the kcat of aminoacylation. The peptide region 583-GNLVNR-588 is highly conserved in all MetRS so far sequenced. We therefore propose that the hydrogen donor/acceptor amino acid residues within this region are the most critical protein determinants for the positive selection of the methionine tRNAs.

Amino Acid Sequence↗

Human amelogenesis. I: High resolution electron microscopy study of ribbon-like crystals.

Ribbon-like crystals, from developing enamel of human fetuses, were studied by high resolution electron microscopy. These crystals were classically described as the first organized mineral formed during amelogenesis. They were characterized by a mean width-to-thickness ratio (W.T-1) of 9.5, and 40% were bent. On lattice images we noted the presence of the central dark line (CDL) associated with white spots. Both structures were found in crystals with a minimum thickness of 8-10 nm. CDLs were localized in the center of the crystals and seemed to be linked to the initial growth process, but their exact structure and function were not fully determined. We were able to study the structure of the ribbon-like crystals with a Scherzer resolution close to 0.2 nm. The good correspondence between experimental and computed images showed that their structure was related to hydroxyapatite (HA). In addition, the presence of ionic substitutions and deficiencies were also compatible with HA. In this study, about 50% of the crystals showed structural defects. Screw dislocations were the most often noted defects and were observed within crystals aligned along five different zone axes. Low- and high-angle boundaries were also detected. Low-angle boundaries, found in the center of the crystals, could thus be related to CDLs and be implicated in the nucleation step of crystal formation, whereas high-angle boundaries could result from the fusion of ribbon-like crystals. Such mechanisms could induce an acceleration of the growth in thickness of the crystal observed during the maturation stage of amelogenesis.

Amelogenesis↗

Identification of potential amino acid residues supporting anticodon recognition in yeast methionyl-tRNA synthetase.

Sequence comparisons among methionyl-tRNA synthetases from different organisms reveal only one block of homology beyond the last beta strand of the mononucleotide fold. We have introduced a series of semi-conservative amino acid replacements in the conserved motif of yeast methionyl-tRNA synthetase. The results indicate that replacements of two polar residues (Asn584 and Arg588) affected specifically the aminoacylation reaction. The location of these residues in the tertiary structure of the enzyme is compatible with a direct interaction of the amino acid side-chains with the tRNA anticodon.

Amino Acid Sequence↗

HREM study of irradiation damage in human dental enamel crystals.

Several phenomena have been observed during the examination of human dental enamel crystals (mainly constituted by hydroxyapatite (OHAP] by high-resolution electron microscopy (HREM) at 300 and 400 keV: orientation-dependent damage in the form of mass loss from voids or uniform destruction of crystal structure, beam-induced diffusion creating outgrowths at the crystal surfaces, recrystallization of the bulk crystal and crystallization of the inorganic components of the matrix surrounding the crystals. These beam-induced crystals have the CaO structure. The phenomena observed are most likely due to various electron-crystal interaction mechanisms (ballistic knock-on damage, electronic excitations, temperature rise, etc.). In this paper, the contribution of the ballistic process to the phenomena observed is discussed. The quantitative description of the knock-on collisions rests on the McKinley-Feshbach cross-section formula. The minimum ion displacement energies which appear in this expression have been estimated on the basis of the electrostatic ion binding energies, and the covalent bond energies if required. It is shown that hydroxyl, calcium and oxygen ions can effectively be displaced by the incident 300 and 400 keV electrons. Thus, the formation of CaO crystals by the combination of calcium and oxygen ions diffusing from their initial sites inside the OHAP lattice can tentatively be explained.

Adult↗

Hybridization studies of poly A-RNA from 5'-bromodeoxyuridine treated neuroblastoma cells.

Hybridization studies were carried out to measure sequence complexity and relative complexity of poly A-RNA populations from M1 neuroblastoma cells cultivated under proliferating conditions and after BrdU treatment. BrdU treatment is known to induce morphological differentiation. Hybridization kinetics were performed with [3H] labelled complementary DNA synthetized by reverse transcriptase action. The total complexities and the complexities of three classes of sequences measured for the two developmental states differed significantly. In particular, the total complexities as well as the complexity of the rare sequences class were higher in the poly A-RNA population of morphological differentiated M1 cells. Heterologous hybridization between poly A-RNA of proliferating cells with cDNA of differentiated M1 cells was very close to the homologous hybridization of poly A-RNA and cDNA from differentiated cells, nevertheless significant differences, were found in the intermediate and in the rare sequences classes. On the other hand the inverse heterologous hybridization (poly A-RNA of differentiated state X cDNA of proliferating cells) showed a lower hybridization in the region of Rot higher than 1. The plateau reached only 87 per cent compared to that of the homologous hybridization, suggesting that certain sequences expressed in the differentiated state. Nevertheless the number of different poly A-RNA species present per cell (seen by homologous hybridization experiments) was higher in differentiated state indicating that selective transcription took place beside repression with morphological differentiation.

Animals↗

Cell interactions with polyelectrolyte multilayer films.

The short-term interactions of chondrosarcoma cells with polyelectrolyte multilayer films built up by the alternate adsorption of poly(L-lysine) (PLL) and poly(L-glutamic acid) (PGA) was studied in the presence and in the absence of serum. The films and their interaction with serum proteins were first characterized by means of optical waveguide lightmode spectroscopy, quartz crystal microbalance, and zeta potential measurements. In a serum-containing medium, the detachment forces measured by the micropipet technique were about eight times smaller on PGA-ending than on PLL-ending films. For these latter ones, the adhesion force decreased when the film thickness increased. In a serum-free medium, the differences between the negative- and positive-ending films were enhanced: adhesion forces on PLL-ending films were 40-100% higher, whereas no cellular adherence was found on PGA-terminating films. PGA-ending films were found to prevent the adsorption of serum proteins, whereas important protein adsorption was always observed on PLL-ending films. These results show how cell interactions with polyelectrolyte films can be tuned by the type of the outermost layer, the presence of proteins, and the number of layers in the film.

Blood Proteins↗

Scoring mitotic activity in longitudinal sections of mouse embryonic incisors: significant differences exist for labial and lingual inner dental epithelia.

The incisors of rodents (e.g., mice and rats) have an enamel-free lingual surface. It is not known how position specific differential evolution of labial versus lingual inner dental epithelium (IDE) is regulated. The patterns of mitotic activity of lingual and labial preodontoblasts and IDE were determined on sagittal sections of embryonic mouse incisors. No statistical differences were observed for the preodontoblasts. On the other hand the lingual IDE withdraw from the cell cycle before the corresponding labial cells. The incapacity of lingual IDE to differentiate into functional ameloblasts might be a consequence of a failure of cell kinetic dependent acquisition of competence to respond to specific epigenetic control mechanisms.

Animals↗

Red blood cell adhesion on a solid/liquid interface: comparison of two models.

Discoid red blood cells (RBCs) deposited irreversibly on a horizontal glass surface are studied by means of optical microscopy and image analysis. The relative surface covered by the RBCs, as well as the variance of this surface coverage as a function of the cell concentration, are analyzed and compared to the results derived from the ballistic deposition (BD) model. This model describes the irreversible deposition of spherical particles under the influence of an infinitely large gravitation force and does not allow for overlaps between adsorbed particles. In spite of these characteristics, the BD model permits, surprisingly, to reproduce our experimental observations on the deposition of RBCs on a flat surface. This finding is discussed, in particular in respect to a former study where a model was developed for colloidal particles of this particular geometric shape.

Cell Adhesion↗