Autoradiographic studies of glial proliferation in different areas of the brain of the 14-day-old rat

Cell cycle parameters, as well as the mode of proliferation of glial cells, in four different areas of the brain of the 14-day-old rat (cortex, corpus callosum, nucleus caudatus putamen and commissura anterior) were studied using different cell kinetic methods after injection of [3H]TdR and/or [14C]TdR. The duration of the S phase (tS) was found to be about 10 hr and that of the cycle time (tC) about 20 hr, tG2 is less than 2 hr and t(G2 + M) about 4 hr. These values are valid for glial cells in all four brain areas studied. However, the labelling index (LI) of the glial cells differs by a factor of 3, between 1.8 and 5.4% in the different brain areas. Accordingly, the growth fraction of the glial cell population in the four areas varies between 0.04 and 0.12. Glial cells (astrocytes as well as oligodendrocytes) proliferate according to a steady state system. Furthermore, the proliferation of glial cells is associated with continuous cell loss. After each mitosis about 3% of the daughter cells become pyknotic and die. In addition, a permanent exchange of glial cells occurs between the proliferating and non-proliferating pool.     

Biosynthesis of 14C- and 35S-lincomycin using different sources for the label]

The sources of radioactive labels were chosen for biosynthesis of labeled lincomycin. The levels of the label incorporation into lincomycin were high with all the sources used when the lincomycin-producing organism was cultivated on the synthetic medium as compared to the complex medium. Incorporation of the label into lincomycin was most effective when I14C- or 214C-thyrosine was used as the precursors. These precursors were effective in both the complex and the synthetic media. In production of significant amounts of the labeled lincomycin I14C- or 214C-sodium propionate, I14C- or 142C-sodium acetate and 14C-protein hydrolysate may be used as the sources of the radioactive carbon. In production of 35S-lincomycin K235SO4 may be used as the source of the label. The optimal conditions for biosynthesis of 14C- and 35S-lincomycin were developed (concentration of some components of the medium, time of the label addition and others).     

The pick-up of C14-DDT at different temperatures

Mosquito larvae, Aedes aegypti, picked up progressively greater amounts of C¹⁴ labeled DDT with ascending temperatures. The pick-up relationship contrasted with per cent mortality which showed a negative temperature coefficient. The exposure temperatures were 10°, 20° and 30° C. A similar relationship of pick-up of DDT occurred when heads, thoraces, or abdomens of the larvae were compared separately. The thorax contained greater concentrations than heads or abdomens at 30° and 20° C. At 10° C the heads contain more than thoraces and abdomens, but the amount was still less than that picked up at 20° and 30° C.The results show a positive coefficient of pick-up of DDT but a negative temperature coefficient for per cent mortality provided the concentration of DDT is not too high. The latter relationship agrees with considerable earlier research. The explanation for the negative temperature effect of DDT is still not understood, but we have evidence that the effect is not positively related to pick-up by whole larvae or portions of whole larvae.

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Zusammenfassung Mückenlarven (Aedes aegypti) nahmen um so größere Mengen von ¹⁴C-markiertem DDT auf, je mehr die Temperatur anstieg. Im Gegensatz dazu zeigte die prozentuale Mortalität einen negativen Temperaturkoeffiziente. Die Untersuchungstemperaturen betrugen 10, 20 und 30°. Eine ähnliche DDT-Aufnahme-Beziehung ergab sich, wenn Köpfe, Brustabschnitte oder Abdomina der Larven getrennt verglichen wurden. Bei 30 und 20° wiesen die Brustabschnitte größere Konzentrationen auf als die Köpfe und Abdomina. Bei 10° enthielten die Köpfe mehr als die Abdomina und Brustabschnitte, aber die aufgenommene Menge war doch geringer als bei 20 und 30°.Die Ergebnisse zeigen einen positiven Temperaturkoeffizienten für die DDT-Aufnahme, aber einen negativen für die prozentuale Mortalität unter der Voraussetzung, daß die DDT-konzentration nicht zu hoch ist. Die zweite Beziehung stimmt mit beträchtlich früheren Untersuchungen überein. Eine Erklärung für diesen negativen Temperatureffekt des DDT ist noch nicht gefunden, aber es scheint, daß diese Wirkung keine positive Korrelation zur DDT-Aufnahme ganzer Larven oder bei Teilen ganzer Larven aufweist.

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Paper No. 5188 Scientific Journal Series, Minnesota Agricultural Experiment Station, St. Paul 1, Minnesota.     

CD14 gene promoter polymorphism in different clinical forms of tuberculosis

Mycobacterium tuberculosis interacts with monocyte-macrophages through cell surface molecules including CD14. A soluble form of CD14 (sCD14) exists in human serum, and higher amounts of it are found in tuberculosis. A polymorphism on CD14 gene promoter was associated with increased sCD14 levels in some diseases. To evaluate whether this polymorphism associates with tuberculosis, its clinical forms, and increased sCD14, genotype/allele frequencies in tuberculosis patients were compared with the controls. Results confirmed increased levels of sCD14 in patients with tuberculosis, and those with miliary tuberculosis had the highest levels. sCD14 decreased to normal levels after anti-tuberculosis treatment. No association was found between the CD14 polymorphism and tuberculosis or sCD14 levels. Results suggest that sCD14 may be involved in anti-tuberculosis immune response, but its increase is a consequence of infection rather than a predisposed genetic trait. Measuring sCD14 in tuberculosis may help monitor anti-tuberculosis treatment.     

Biosynthesis of the labelled rifamycin B in the presence of different 14C- and 3H-precursors

The results of studying the effectiveness of incorporation of the label from different 14C- and 3H-precursors into the molecule of rifamicin B during its biosynthesis are presented. The regularities of the label incorporation into the antibiotic composition as dependent on the time of the precursor addition were investigated. A radiochemically pure preparation of 14C-rifamicin B with specific radioactivity of 3 mcurie/mg was obtained.     

Characterization of 14 different putative protein kinase cDNA clones of the C4 plantSorghum bicolor

C₄ photosynthesis is functionally dependent on metabolic interactions between mesophyll- and bundle-sheath cells. Although the C₄ cycle is biochemically well understood, many aspects of the regulation of enzyme activities, gene expression and cell differentiation are elusive. Protein kinases are likely involved in these regulatory processes, providing links to hormonal, metabolic and developmental signal-transduction pathways. Here we describe the cloning and characterization of 14 different putative protein kinase leaf cDNA clones from the C₄ plant Sorghum bicolor. These genes belong to three different protein kinase subfamilies: ribosomal protein S6 kinases, SNF1-like protein kinases, and receptor-like protein kinases. We report the partial cDNA sequences, mesophyll/bundle-sheath steady-state mRNA ratios, mesophyll/etiolated leaf steady-state mRNA ratios, and the positions of 14 protein kinase genes on the genetic map of S. bicolor. Only three of the protein kinase genes described here are expressed preferentially in mesophyll cells as compared with the bundle-sheath.     

14C-labelling patterns of phytoplankton: specific activity of different product pools

Primary production, release of extracellular products (EOC)and the distribution of ¹⁴C into primary products of photosynthesis(low mol. wt. products, lipids, polysaccarides, and proteins)were investigated during 2 months in spring 1983 in eutrophicLake Hylke, Denmark. After half days in situ incubations, eachfraction was assayed for ¹⁴C activity and the standing stockof carbon. Calculation of specific activity revealed non-equilibriumconditions in all fractions. Since the intracellular fractionsact as precursors of EOC, this means that the release may beseriously underestimated (3–20 times) by assuming isotopicequilibrium between the inorganic carbon pool and the EOC productsin the Extracellular medium. However, measurements of the standingstock of carbon are complicated by inclusion of non-phytoplanktonmaterial, thereby making calculation of specific activity uncertain. Present address: Carbon 14 Centralen, The International Agencyfor ¹⁴C Determination, 11, Agern Allé, DK-2970 Hørsholm,Denmark     

The chromosome breakpoint at 14q32 in an ataxia telangiectasia t(14;14) T cell clone is different from the 14q32 breakpoint in Burkitts and an inv(14) T cell lymphoma

Summary The T cell receptor chain gene locus and the immunoglobulin heavy chain gene locus (IgH) have previously been mapped to the q11 and q32 positions respectively of the human chromosome 14. Both of these sites are also common breakpoints in lymphocytes from ataxia telangiectasia (A-T) patients. Using in situ hybridisation we show that the 14q32 breakpoint in an A-t non-leukaemic T cell clone with t(14;14) translocation, lies outside the IgH locus and proximal to it with respect to the centromere. The 14q11-14qter segment of the homologous chromosome 14 carrying the constant gene region of the chain locus is translocated to this 14q32 position.     

A chromosome 14-specific human satellite III DNA subfamily that shows variable presence on different chromosomes 14.

We describe a new subfamily of satellite III DNA (pTRS-63), which, by a combination of in situ hybridization to human metaphase chromosomes and analysis of a panel of somatic cell hybrids, is shown to be specific for human chromosome 14. This DNA has a basic 5-bp repeating unit of diverged GGAAT which is tandemly repeated and organized into either one of two distinct higher-order structures of 5 kb (designated the "L" form) or 4.8 kb (designated the "S" form). In addition, a third (Z) form, representing no detectable levels of this satellite III subfamily, is found. Results from five somatic cell hybrid lines and from a number of informative human individuals suggest that, on any one chromosome 14, only one of the three forms may exist. Subchromosomally, this sequence has been mapped to the p11 region and is distal to the domain occupied by another previously described satellite III subfamily (pTRS-47) found on chromosome 14. The pTRS-63 sequence described adds to the understanding of the structural organization of the short arm of human chromosome 14 and should be useful for the investigation of the molecular etiology of the frequently occurring t(13q14q) and t(14q21q) Robertsonian translocations.     

Transport of14C-IAA from leaves and shoots to different fruit parts

Transport of¹⁴C-IAA was studied in apple spurs of a 20-year-old McIntosh with one fruit and one shoot. Water solutions of IAA were applied to intact, pricked or scratched leaf blades, to decapitated shoots or to petioles (leaf-blade removed) at the end of June, July and August.¹⁴C-IAA (in an unknown form) was transported from intact leaves and shoots to pedicel, pericarp and seeds. Radioactivity of the pedicels increased every month while that of seeds reached maximum at the end of July and then markedly decreased in August. Total radioactivity of whole fruit doubled, at least, with every month due to enlargement of the pericarp. Pedicels deprived of fruits had their retention prolonged on spurs with leaves or shoots treated with 1% IAA in lanoline. It is assumed that auxin delivered from shoots or still growing leaves at the time of its deficiency in seeds, restrains fruits from premature dropping. At the same time seeds seem to be protected by a regulatory system in pedicel against too massive flow of auxin from outside.     

Surfactant proteins A and D bind CD14 by different mechanisms

Surfactant proteins A (SP-A) and D (SP-D) are lung collectins that are constituents of the innate immune system of the lung. Recent evidence (Sano, H., Sohma, H., Muta, T., Nomura, S., Voelker, D. R., and Kuroki, Y. (1999) J. Immunol. 163, 387-395) demonstrates that SP-A modulates lipopolysaccharide (LPS)-induced cellular responses by direct interaction with CD14. In this report we examined the structural elements of the lung collectins involved in CD14 recognition and the consequences for CD14/LPS interaction. Rat SP-A and SP-D bound CD14 in a concentration-dependent manner. Mannose and EDTA inhibited SP-D binding to CD14 but did not decrease SP-A binding. The SP-A binding to CD14 was completely blocked by a monoclonal antibody that binds to the SP-A neck domain but only partially blocked by an antibody that binds to the SP-A lectin domain. SP-A but not SP-D bound to deglycosylated CD14. SP-D decreased CD14 binding to both smooth and rough LPS, whereas SP-A enhanced CD14 binding to rough LPS and inhibited binding to smooth LPS. SP-A also altered the migration profile of LPS on a sucrose density gradient in the presence of CD14. From these results, we conclude that 1) lung collectins bind CD14, 2) the SP-A neck domain and SP-D lectin domain participate in CD14 binding, 3) SP-A recognizes a peptide component and SP-D recognizes a carbohydrate moiety of CD14, and 4) lung collectins alter LPS/CD14 interactions.     

不同年份生产的乙型脑炎减毒活疫苗SA14-14-2基因稳定性研究

为了研究不同年份生产的乙型脑炎减毒活疫苗病毒E蛋白基因稳定性,从分子水平控制乙型脑炎减毒活疫苗质量,确保疫苗安全性,本研究分析了不同年份生产的乙脑活疫苗病毒E蛋白基因核苷酸序列及编码的氨基酸序列,并与该疫苗原始种子、主种子、工作种子、乙脑病毒强、弱毒株进行比较。结果显示不同年份生产的乙脑活疫苗病毒E蛋白基因核苷酸序列与其原始种子、主种子、工作种子和基因库中登录的乙脑病毒弱毒株SA14-14-2的相应序列完全一致,与乙脑病毒强毒株SA14的E蛋白氨基酸序列比较有9个位点氨基酸发生了改变。不同年份生产的乙脑活疫苗病毒E蛋白基因稳定性表明该疫苗质量稳定、安全。     

Translocation and incorporation of 14C into the petiole from different regions within developing cottonwood leaves

Summary The ability of a developing cottonwood (Populus deltoides Bartr.) leaf to export ¹⁴C-labeled assimilates begins at the lamina tip and progresses basipetally with increasing LPI. This progression indicates that portions of leaves function quasi-independently in their ability to export ¹⁴C-photosynthate. Although most of the exported radioactivity was recovered in the petiole as water-80% alcohol-soluble compounds, there was also substantial incorporation into the chloroform and insoluble fractions. This observation indicates that assimilates translocated from the lamina are used in structural development of the petiole. Freeze substitution and epoxy embedding were used to prepare microautoradiographs for localization of water-soluble compounds. Radioactivity was found in all cell types within specific subsidiary bundles of the petiole. However, radioactive assimilates appeared to move from the translocation pathway in the phloem toward active sinks in the walls of the expanding metaxylem cells. Translocation in the mature xylem vessels was not observed.