Autoradiographic studies on RNA synthesis and transport in the ovary ofHydrophilus olivaceus fabr. (Hydrophilidae,Polyphaga, Coleoptera)

Summary Autoradiographic studies using³H-uridine in the ovary ofHydrophilus olivaceus Fabr. show that nurse cells, the germinal vesicle and follicular nuclei play an important role in contributing RNA whereby the major portion of RNA comes from the nurse cells.I am grateful to Prof. P. S. Ramamurty, School of Life Sciences, University of Hyderabad, Hyderabad, India, for his constant inspiration during the tenure of the present investigation.     

Mechanism of age-dependent involution in embryonic chick notochords

To study the possible mechanism of the age-dependent involution of the notochord, isolated mesenchymefree notochords of chick embryos were cultured in vitro and compared with their counterparts in vivo. Two different aspects were evaluated: (1) DNA synthesis measured by [³H]thymidine incorporation and visualized by autoradiography and (2) cell death quantified by counting the number of pyknotic nuclei. The results demonstrate that [³H]thymidine uptake by notochords shows an age-dependent decrease in vitro as well as in vivo. The number of [³H]thymidine-labelled notochord cells, however, is higher in vitro than in vivo. At the same time, there is an age-dependent increase in pyknosis in the notochord in vivo and in vitro. So, during the aging process, the number of both pyknotic nuclei and of [³H]thymidine-labelled nuclei suggest a high turnover of notochord cells in vitro. From these results, we can conclude that the process of involution in aging notochord seems to be controlled by a programmed intrinsic process, which might be influenced partially by the microenvironment in vivo.     

Migration intercinétique dans la placode olfactive chez l'embryon de poulet interkinetic nuclear migration in nasal placode

Summary Cells of nasal placode of chick embryos were studied with thymidine H³ and autoradiography. Our results shown, that the nuclei in the nasal placode synthesize DNA in the outer zone, then migrate toward the inner zone to undergo division and subsequently return to the outer zone.     

Einbau von H3-thymidin in zellen und gewebe vonHelix pomatia L. (gastropoda,pulmonata)

Summary It has been demonstrated by injections of H³-Thymidine into adult and juvenile snails (Helix pomatia) that the mitotic activity in the central nervous system, the connective tissue and epidermis was generally higher in juvenile animals than in adult ones. Data are given for the amount of triated thymidine labelled nuclei in the tissues, which are investigated.     

In vitro effects of methionine-enkephalin,somatostatin and insulin on cultured gonadal cells of the snailHelix aspersa

Isolated snail gonadal cells were cultured in the presence of synthetic neuropeptides in order to determine the subsequent effect of these substances on gonadal synthetic activities. Gonadal cells were incubated for 24 h in concentrations of methionine-enkephalin, somatostatin and insulin ranging from 10^–4 M to 10^–9 M, in medium 199 supplemented with 6% Ultroser G. Synthesis of DNA and protein by the cultured cells were simultaneously estimated by measuring incorporation of³H thymidine and³⁵S methionine. The rate of labelled precursor incorporation was measured using the liquid scintillation technique. All substances tested exerted a dose-dependent effect. The synthetic activity of the cultured cells was highest when the concentration of the peptides added to the medium approximated the physiological levels. Methionine-enkephalin, somatostatin and insulin at 2×10^–8 M significantly increased³H thymidine incorporation, by 62%, 69% and 69% respectively, and protein synthesis by 42%, 57% and 57%, respectively. In the case of juvenile gonadal cultured cells, a similar increase in³H and³⁵S incorporation was registered for a 10^–7 M peptide concentration. Both lower and higher peptide concentrations inhibited³H thymidine and³⁵S methionine incorporation. Pharmacological studies suggest the existence of methionine-enkephalin and somatostatin-like receptors on snail gonadal cells. These results indicate that our gonadal cell culture model provides a useful tool for the study of the neuroendocrinological control of the activity of snail gonadal cells.     

Aufnahme von thymidin durch mastocytomzellenin vitro unter der einwirkung von podophyllumstoffen

Summary SP-G and SP-I, two podophyllum preparations with spindle poison activity, were tested for their effect on³H-thymidine incorporation into P-815 mastocytoma cellsin vitro. Concentrations which inhibited cell multiplication completely did not affect DNA-synthesis as measured by thymidine incorporation.     

A chromosome study of the parthenogenetic rice water weevil,Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae), in Japan

Summary The chromosomes of the so-called rice water weevil,Lissorhoptrus oryzophilus, Kuschel obtained from Tokoname City, Aichi Prefecture, were studied in both oogonial and oocyte-maturation divisions in squash and sectioned slides, respectively. The chromosome number was confirmed as exactly 33 in both divisions. No reduction division takes place. It is therefore concluded thatLissorhoptrus oryzophilus is a parthenogenetic triploid in Tokoname City.     

Über den Einbau14C-markierter Aminosäuren in Zellkernfraktionen aus Meerschweinchenleberzellenin vitro

Summary The incorporation of¹⁴C-amino acids into ribonucleoproteid particles obtained by differential centrifugation of mechanically disintegrated liver cell nuclei was demonstratedin vitro. Nuclear supernatant revealed also very remarkable incorporation.     

Etude autoradiographique de l'incorporation de la thymidine-3H,de l'uridine-3H et de la phénylalanine-3H dans les chloroplastes deClivia miniata etBilbergia sp.

Summary Mature chloroplasts ofClivia miniata etBilbergia sp. show the intensive incorporation of thymidine³H into DNA and uridine³H into RNA. On the contrary, phenylalanine³H is mostly incorporated in the proplastids.     

Effect of tingenone,a quinonoid triterpene,on growth and macromolecule biosynthesis inTrypanosoma cruzi

Summary Tingenone and horminone, two natural quinonoid substances, inhibited the in vitro growth ofTrypanosoma cruzi, 30 M drug concentration producing total inhibition of growth. Tingenone inhibited total uptake and incorporation of [³H]thymidine, [³H]uridine, L-[³H]leucine into parasite macromolecules. Other quinonoids assayed were either less effective (abruquinone A) or even quite inactive (visminone B and ferruginin B). Investigation of several mechanisms for the cytotoxic action of tingenone pointed to the interaction with DNA as the most likely factor involved. Tingenone also inhibited the growth ofCrithidia fasciculata, but the drug was significantly less active on this organism than onT. cruzi.This work was supported by grants of UNDP/World Bank/World Health Organization Special Programme for Research and Training in Tropical Diseases, Organization of American States (Multinational Programme of Biochemistry) and Programa Nacional de Enfermedades Endémicas (SECYT), República Argentina. A preliminary account was given at the Workshop on Oxidative Damage and Related Enzymes, Frascatti (Italy), 1983.     

Zum Proliferations-Stoffwechsel der Rattenleber nach Teilhepatektomie. Autoradiographische Untersuchungen mit3H-Cytidin,-Phenylalanin und -Thymidin

Summary During the first wave of parenchymal liver regeneration in adult rats after partial hepatectomy, the cellular synthesis and migration of RNA and the metabolism of protein were studied by autoradiography following an injection of³H-cytidine or³H-l-phenylalanine and double injections of 1 of these precursors +³H-thymidine. The following results were obtained: the synthesis and migration of RNA and the metabolism of protein are enhanced under these conditions of proliferation. In spite of this, the relation of metabolic activity in nucleolus, karyoplasm and cytoplasm remains constant. By double injection techniques it is proved that no differences exist in migration of RNA into the cytoplasm and cytoplasmic protein synthesis between cells with or without DNA synthesizing nuclei.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Centromeric heterochromatin in the karyotype of the male Greater Kudu (Tragelaphus strepsiceros)

Summary The chromosomes of a male Kudu (Tragelaphus strepsiceros) have been studied by C-banding and H³ thymidine labelling. It is suggested that heterochromatin may have accumulated on the 14th pair of autosomes before its translocation to the Y-chromosome.     

Production of insulin-like growth factor I (IGF-I) and IGF-binding proteins by rat intestinal stromal cells in vitro

To determine if intestinal stromal cells secrete diffusible factors such as insulin-like growth factors (IGFs) capable of regulating epithelial cell growth in vitro, stromal cells were isolated by enzymatic digestion of rat intestine. Incorporation of [³H]thymidine into DNA and [¹⁴C]leucine into protein of IEC-6 cells, a model intestinal epithelial cell line, was significantly increased (two- to threefold) when the IEC-6 cells were co-cultured with stromal cells, relative to IEC-6 cells grown alone. Medium conditioned by stromal cells stimulated DNA synthesis of IEC-6 cells in a dose-dependent manner. Analysis of the conditioned medium revealed that intestinal stromal cells secreted IGF-I, but little IGF-II, in addition to an M _r 32,000 IGF-binding protein (IGFBP-2) and an IGFBP having M _r∼ 24,000. We conclude that rat intestinal stromal cells secrete one or more diffusible factors, which may include IGF-I and IGFBPs, capable of stimulating proliferation of IEC-6 cells in vitro. Received 25 August 1997; received after revision 7 November 1997; accepted 20 November 1997     

Nucleolus DNA synthesis inVicia faba root-tip meristems

Summary Autoradiographs ofVicia faba lateral root-tip meristems were prepared following a 30 min pulse with³H-thymidine. 1/3 of all interphase nuclei in the meristem are labeled, most with a uniform distribution of silver grains. 2–5% of labeled nuclei show specific nucleolus DNA labeling.     

Progesterone inhibits human endothelial cell proliferation through a p53-dependent pathway

Previous studies have shown that progesterone inhibits endothelial cell proliferation through a nuclear receptor-mediated mechanism. Here, we further demonstrate that progesterone at physiologic levels (5 – 500 nM) dose- and time-dependently inhibited DNA synthesis of cultured human umbilical vein endothelial cells (HUVEC). The mRNA and protein levels of p21, p27, and p53 in HUVEC were increased by progesterone. The formation of CDK2-p21 and CDK2-p27 were increased and the CDK2 activity was decreased in the progesterone-treated HUVEC. The progesterone-inhibited [3H]thymidine incorporation was completely blocked when the expressions of p21 and p27 were knocked-down together. Transfection of HUVEC with dominant negative p53 cDNA prevented the progesterone-induced increases in p21 and p27 promoter activity and protein level, decreases in thymidine incorporation, and capillary-like tube formation. Matrigel angiogenesis assay in mice demonstrated the antiangiogenic effect of progesterone in vivo. These findings demonstrate for the first time that progesterone inhibited endothelial cell proliferation through a p53-dependent pathway. Received 28 July 2008; received after revision 25 September 2008; accepted 26 September 2008     

Application de méthodes d'analyse biochimique à une étude taxonomique: les corégones du lac de Neuchâtel

Summary Two closely related forms ofCoregonus from Lake Neuchatel were examined cytologically and biochemically, in order to ascertain the chromosome number and the DNA content of haploid and diploid nuclei.Coregonus fera has 2N=78 ± 2 chromosomes, and a DNA content (diploid) of 5.8 × 10⁻⁹ mg;Coregonus macrophthalmus, 2N=78⁺ ± 3, DNA content of 6.1 × 10⁻⁹ mg. The difference between the two DNA constants is statistically significant. These results do not support the hypothesis which postulates that polyploidy may be a determining factor in the speciation of these fishes.      

DNS-Synthese,Blastentransformation und Mitosen in gemischten,homologen Granulozyten-Lymphozytenkulturen

Summary Lymphocyte stimulation in vitro could be obtained with allogeneic but not autochthonous or isogeneic granulocytes. Blastoid transformation, mitosis and activated DNA synthesis measured by H³-thymidine incorporation occurred simultaneously. The cytoimmunological relevance and further applications of the findings are discussed.

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Mit Unterstützung der schweizerischen Krebsliga.     

Zytologische Untersuchungen an der Reblaus-Blattgalle

Summary Up to 9 types of cell nuclei, distinguished by their size, were found in the tissue of galls, which are produced by the vine louse. The growth of the cell nuclei probably proceeds in a series of prophase-like intermediate stages from one type of nuclei size to the next largest by means of endomitotic or endomitosis-like processes.     

Selective antiproliferative effects of thymidine

A substance with antiproliferative bioactivity in an aqueous extract ofCordyline terminalis was purified and identified by mass spectrometry to be the natural nucleoside, thymidine. 10^–5M Thymidine inhibited EL4 cell replication and decreased cell viability after 12–24 h. The effect was highly specific for this nucleoside. Treated cell cultures showed a significant increase in S phase cells and a corresponding decrease in G₁ phase cells. Nitrobenzylthioinosine (which prevented facilitated entry of thymidine) protected cells from the antiproliferative action of thymidine. A human breast cancer cell line (MCF7) was also growth-inhibited by 10^–5M thymidine but a murine lymphoma cell line (K36) was not. Thus, submillimolar thymidine has effects on cell proliferation which are selective both with respect to specificity for the compound and for different tumour cell lines.     

Einbau von H3-d-glucose während der Oogenese beiApis mellifica L.

Summary The incorporation of tritiated glucose into the ovary of honey bee queens was studied by autoradiography. During the last stages of oogenesis, the synthesis of respectable amounts of glycogen was found in the reticuloplasm of the developing eggs. The follicle epithelium and also the nurse cells only appeared more lightly and transitory labelled at medium stages. An hypothesis was established concerning an antagonism of protein and polysaccharide formation in the cytoplasm corresponding to the rather late occurrence of glycogen in insect oogenesis.