The effect of proteoglycans of cartilage and over-sulphated polysaccharides on the development of calcium-hydroxy-apatite (CHA) crystal formation in vitro

A coulometric model system is described which facilitates the quantitative study of the kinetics of transformation of amorphous calcium phosphate (ACP) into calcium-hydroxy-apatite (CHA) crystals. Proteoglycans of high molecular weight and over-sulphated polysaccharides (Arteparon, dextran sulphate) delayed CHA crystal formation. The results have enabled us to characterize the structure activity relationship of inhibitors of CHA formation, and to postulate a general structural requirement for molecules with inhibitory effect. As working mechanism, binding of calcium ions by sulphate groups of polyanions was supposed, which might reversibly impair "the critical nuclei formation", and/or further deposition of calcium ions in the CHA crystals. The clinical, therapeutical significance of the determination of the threshold concentration of different compounds is discussed.     

The effect of perinatal oestrogen treatment on the troph-hormone secreting cells of the anterior pituitary of the rat

Late effect of the perinatal administration of oestradiol dipropionate on pituitary gonadotrophs and the morphology of the LH-RH neuronal system was tested both in male and female rats. Oestrogen caused a severe reduction in the number and size of immunodetectable gonadotrophs in both sexes. By the 90th day of life, however, immunomorphology and distribution of the gonadotroph cells had became normal, and also the LH-RH system of the animals was similar to that of the intact controls. The lack of vaginal cycles indicated, however, that oestrogen might have permanently impaired higher brain centers regulating cyclic gonadotroph hormone release.     

Cytogenetics of the chinese giant salamander,Andrias davidianus (Blanchard): the evolutionary significance of cryptobranchoid karyotypes

Cytogenetic aspects of the cryptobranchid salamander Andrias davidianus of western China have been studied, including chromosome number and morphology, C-band patterns, meiosis, and the chromosomal localization of ribosomal 5S RNA genes. Our data regarding chromosome number (2n=60) and general chromosome morphology largely confirm the results of Morescalchi et al. (1977). The karyotype consists of 16 pairs of macrochromosomes that decrease gradually in relative length to 14 pairs of microchromosomes. Telocentric chromosomes are a conspicuous feature of the karyotype, representing more than half the genome. Differential staining reveals that all of the chromosomes, except four pairs of microchromosomes, have C-band heterochromatin in their centromeric regions, the amount varying irrespective of chromosome size. Faint bands of interstitial and telomeric C-band heterochromatin are found in mitotic chromosomes but are not seen in meiotic preparations. In C-banded mitotic preparations from a female, one of the smallest macrochromosome pairs is heteromorphic in respect to C-band heterochromatin and centromere position. In situ hybridization of an iodinated 5S RNA probe to meiotic chromosome preparations reveals that this repeated gene is clustered near the telomeric region of chromosome 7, a medium size telocentric, a location corresponding to a band of heterochromatin. Studies of spermatocytes indicate that the process of meiosis in A. davidianus closely resembles that of more advanced salamanders, and that the microchromosomes are meiotically stable. The significance of microchromosomes and chromosome morphology in the reorganization of salamander genomes during evolution is discussed on the basis of cytogenetic data available for A. davidianus and various other primitive and advanced salamanders.     

Urinary prostaglandin and kallikrein excretion are not flow dependent in the rat

To study the relationship between urine flow, urinary prostaglandin (PG) and kallikrein excretion in the rat high urine flow was induced in hydropenic Long-Evans rats by either hypotonic volume expansion or with manniitol or with furosemide. PGE, excretion remained unchanged during hypotonic volume expansion (134.5 ± 29.7 before and 153.0 ± 48.9 pg/min after) while it decreased significantly with mannitol (from 166.3 ± 32.4 to 45.2 ± 8.2 pg/min, p<0.01) and with furosemide (from 170.0 ± 20.4 to 29.5 ± 5.3 pg/min, p<0.001). PGF_2α excretion rates were slightly reduced following all three interventions. Urinary kallikrein excretion remained unchanged in all three groups of animals. It is concluded that, in contrast to human and dogs in the rat urine flow and urinary PG excretion are not interlinked.     

Dipole moments of random coil polypeptides

The Rotational Isomeric States model is applied to calculate dipole moments of polypeptides of the twenty natural α-amino acids in the random coil state. Dipole moments of each repeat unit (μ_i), are evaluated using a quantum mechanics procedure. Dipole moment ratios ($\text{D}{}_{\mathrm{x}}\text{=}\text{〈μ}{}^2\text{〉}\text{x}\text{μ}{}_{\mathrm{i}}{}^2\text{,}\text{x = number of repeat units}$) of homopolypeptides are calculated and extrapolated to x →?. With a few exceptions, D_? = 0.36 ± 0.1. Ten actual proteins and three enzymes are also studied; their dipole ratios (D_x′ =〈μ〉/x) range from 7.34 to 10.57 in 10^?59 C² m² (6.6–9.5 D²). Diffferences in the values of D_x′ are due mainly to the different contributions, μ_i, of the amino acid residues contained in each polymer, whereas the sequence of amino acids has a very minor effect.     

Induction of ion transport in rat heart mitochondria by fluorescamine

This report describes experimental results which show that the fluorescent reagent fluorescamine induces mitochondrial energy-independent swelling when the incubation media contain the chloride salts of the cations Li+, Na+, K+, Rb+, and Cs+. The reaction depends on the concentration of the dye and is inhibited by Mg2+, and its extension is closely related to the amount of the primary amino groups titrated by fluorescamine. Analysis of the labeled inner membrane in polyacrylamide gel shows that the amount of aminofluorescamine complex is lower when mitochondria are in the presence of Mg2+.     

Discrepancies between flow cytometric analysis and [3H]thymidine incorporation in stimulated lymphocytes

The DNA synthesis system of freshly isolated tonsillar lymphocytes and those stimulated by phytohaemagglutinin were compared by different methods. Both cell populations had high DNA polymerase α and thymidine kinase activities, as well as a high rate of incorporation of [³H]thymidine into DNA. However, the two cell populations differed when their DNA distributions were compared by flow cytometry. Freshly isolated cells contained many less (6%) cells in S phase than were found in phytohaemagglutinin-stimulated lymphocytes (18%) as detected by flow cytometry. The labelling of different subpopulations of lymphocytes was studied by sorting them electrically with a fluorescence-activated cell sorter. Analysis of the radioactivity of [³H]thymidine pulse-labelled cells, sorted according to their DNA content, showed that cells in the G₁ peak of DNA distribution had a significant amount of incorporated [³H]thymidine. Sorting of cells according to their size (i.e., by light scattering) revealed that only large cells were labelled with [³H]thymidine.     

Cell surface antigens reacting with antiretrovirus sera on normal mouse spleen cells: a flow cytometric study

The development of erythroleukemia in Balb/c mice injected with Rauscher leukemia virus has been followed by indirect immunofluorescence technique and flow cytometry, using antiserum against disrupted of virions. The progression of the disease was accompanied by a great increase in the number of large, immunofluorescence positive cells. A subpopulation of normal spleen cells was also highly positive. The expression of the antigens in normal cells was examined in relation to the cell cycle. The majority of the S-G2/M phase cells were found in the antigen positive compartment of larger cells. A two-color analysis of immunofluorescence and DNA content revealed that the distribution of antigen expression in G1 phase was broad, gradually decreasing from a low-intensity mode. The cell with double DNA content distributed evenly around a modus of five-fold higher intensity. In experiments with stimulated bone-marrow cells, superiority of S-phase cells in anti-Rauscher serum binding was found. Cell-surface gp70 antigen is suggested to be involved in this cell-cycle dependent binding of antibodies by normal cells.     

The role of N-acyl groups in the inhibitory activity of LH-RH analogues

Inhibitory analogues of luteinizing hormone-releasing hormone (LH-RH) were prepared with formyl-D-Trp¹, acetyl-D-Trp¹, valeryl-D-Trp¹, tartaryl-D-Trp¹, diacetyl-tartaryl-D-Trp¹, acetyl-Gly¹, and acetyl-Sar¹ successively replacing the position one in the analogue [D-Trp¹, D-p-Cl-Phe², D-Trp³, D-Phe⁶, D-Ala¹⁰]-LH-RH. The formyl-D-Trp¹ and acetyl-D-Trp¹ analogues yielded 100% blockade of ovulation at the 10 μg dose; the others were less potent and inhibited ovulation at the 50 μg dose. The inhibitory potency seems to correlate with the polarity of the acyl group.     

Distribution of ABO blood groups and other genetic markers in mothers of infants with congenital malformations

The distribution of ABO and Rh (antigen D) blood groups and of serum albumin, haptoglobin and transferrin variants, in a group of mothers of malformed newborns was investigated. In the first phase of the study, the results showed borderline statistical differences in the distribution of the transferrin types between the study group and a suitable control population. The second phase of the research, where only transferrin phenotypes were studied, showed the same trend as in the first one, but the results were not statistically significant. We conclude that probably there are no true distribution differences, but that it would be desirable to study this problem in a different ethnic group.