Accessibility of host cell lineages to medaka stem cells depends on genetic background and irradiation of recipient embryos

Chimera formation is a powerful tool for analyzing pluripotency in vivo. It has been widely accepted that host cell lineages are generally accessible to embryonic stem (ES) cells with the actual contribution depending solely on the intrinsic pluripotency of transplanted donor cells. Here, we show in the fish medaka (Oryzias latipes) that the host accessibility to ES cell contribution exhibits dramatic differences. Specifically, of three albino host strains tested (i ¹ , i ³ and af), only strain i ¹ generated pigmented chimeras. Strikingly, this accessibility is completely lost in i ¹ but acquired in i ³ after host γ-irradiation. Host irradiation also differentially affected ES cell contribution to somatic organs and gonad. Therefore, the accessibility of various host cell lineages can vary considerably depending on host strains and cell lineages as well as on irradiation. Our findings underscore the importance of host genotypes for interpreting donor cell pluripotency and for improving ES-derived chimera production.     

Die Einlagerung von radioaktivem Schwefel in die Flügel von Acrididen nach Applikation von35S-Natriumsulfat-und35S-l-Cystinlösungen

Summary Psophus stridulus L. andOedipoda coerulescens L. were fed or injected with solutions of Na ₂ ³⁵ SO₄ and³⁵S-l-cystine. The radioactive radiation pattern of the wings was found to depend on the time of application. Differences in radiation intensity were found to correspond to the red, blue and dark areas.     

Dietary vitamin E does not protect from endotoxin-induced hepatic microvascular dysfunction

Starting from the concept that lipopolysaccharide (LPS)-associated hepatotoxicity involves the action of reactive oxygen species, the present study was conducted to test whether vitamin E, a lipophilic antioxidant, prevents LPS-induced hepatic microvascular dysfunction and liver injury. Fifty-two rats were divided into three groups and fed diets containing 0 (n=16), 75 (n=18) or 8000 mg (n=18) α-tocopherol acetate/kg food for four weeks. At 1 h and 6 h after intravenous LPS-exposure (10 mg/kg E. coli LPS) hepatic microvascular response and liver injury were assessed by the analysis of Kupffer cell phagocytic activity, leukocyte-endothelial cell interaction and nutritive sinusoidal perfusion (intravital fluorescence epi- illumination technique) as well as bile flow, serum liver enzyme activities and tissue histomorphology. In animals fed with 75 mg vitamin E/kg (standard diet), LPS caused hepatic Kupffer cell activation (increased phagocytic activity) and hepatic microvascular leukocyte activation, with stasis in sinusoids and adherence in postsinusoidal venules (1 h) followed by leukocytic infiltration into tissue (6 h) and progredient sinusoidal perfusion failure (6 h). Hepatic microvascular injury was accompanied by reduced bile flow and enhanced liver enzyme release. Vitamin E-enriched diet (8000 mg/kg) and even vitamin E-deficient diet did not significantly affect LPS-induced hepatic microvascular cell activation and perfusion failure. Thus, we conclude, that vitamin E is not effective to protect from endotoxin-induced hepatic microvascular dysfunction. Received 7 November 1996; received after revision 30 December 1996; accepted 20 January 1997     

The ovicidal activity ofcis-dehydromatricaria ester: time-dependance of its enhancement by UV light

Summary Freshly laid eggs ofDrosophila melanogaster were treated with the polyacetylenic compoundcis-dehydromatricaria ester. The toxicity of the chemical was enhanced by treatments with long-wavelength ultraviolet light, and the maximum effect was observed when the UV irradiation was performed 4–5 h after the initial contact of the eggs with the chemical.We are grateful to Professor E. Spiess for advice and assistance, and to the National Institutes of Health (GM 24144) for financial support.     

Isomériecis-trans des ménaquinones et oxydations phosphorylantes dans les extraits deMycobacterium phlei

Summary Incubation ofM. phlei washed cells with [¹⁴C³H₃]-l-methionine led to [2-¹⁴C³H₃] dihydromena-quinone-9 with an isotope ratio identical to that of methionine. Chromatography of the doubly labelled quinone indicated, despite a pronounced isotope effect, that bothcis andtrans isomers had the same isotope ratio. This result eliminates any possibility of hydrogen exchange in the 2-methyl group of menaquinones during oxydative phosphorylation, even in thecis isomer. Furthermore, it is confirmed that this compound is certainly formed from natural or synthetic menaquinones during isolation or incubation periods by the effect of daylight irradiation.     

Regulation of the cell cycle following DNA damage in normal and Ataxia telangiectasia cells

A proportion of the population is exposed to acute doses of ionizing radiation through medical treatment or occupational accidents, with little knowledge of the immedate effects. At the cellular level, ionizing radiation leads to the activation of a genetic program which enables the cell to increase its chances of survival and to minimize detrimental manifestations of radiation damage. Cytotoxic stress due to ionizing radiation causes genetic instability, alterations in the cell cycle, apoptosis, or necrosis. Alterations in the G1, S and G2 phases of the cell cycle coincide with improved survival and genome stability. The main cellular factors which are activated by DNA damage and interfere with the cell cycle controls are: p53, delaying the transition through the G1-S boundary; p21^WAF1/CIPI, preventing the entrance into S-phase; proliferating cell nuclear antigen (PCNA) and replication protein A (RPA), blocking DNA replication; and the p53 variant protein p53as together with the retinoblastoma protein (Rb), with less defined functions during the G2 phase of the cell cycle. By comparing a variety of radioresistant cell lines derived from radiosensitive ataxia talangiectasia cells with the parental cells, some essential mechanisms that allow cells to gain radioresistance have been identified. The results so far emphasise the importance of an adequate delay in the transition from G2 to M and the inhibition of DNA replication in the regulation of the cell cycle after exposure to ionizing radiation.     

Increased ALA dehydratase activity and spleen weight in lead-intoxicated rats. A consequence of increased blood cell destruction

Summary Lead was given in the diet (1%) to rats from birth and at different times the animals were studied for delta amino levulinic acid dehydratase (ALAD) activity, spleen weight,⁵⁹Fe incorporation in erythrocytes and⁵¹Cr-labeled erythrocytes survival. The increased ALAD and spleen weight found after lead treatment is explained as a consequence of a shortened survival, which results in a younger age of circulating erythrocytes with higher ALAD activity.Supported by Contract No. 140-76-12 ENV. B. of the E. C. Environmental Research Program.     

Les transformations des nucléoides deEscherichia coli déclenchées par les rayons X

Summary In order to obtain further knowledge on the action of radiation on living cells we studied the nuclear transformations ofE. coli after exposure to X-rays. Three different phenomena could be observed; One is producting the polychromosomal form of nucleoids, and is reversible. The second, which is probably a rapid lethal effect, expresses himself by the vesicular form of the nucleoids. The third is only concerned with very high doses. The hypothesis, that the fragmentation of nucleoids may be directly related to induction of lysogenic cells is postulated.     

Änderung der Wachstumskinetik von Ehrlich-Aszitestumoren verschiedener Ploidie nach Röntgen-Bestrahlung

Summary The effect of irradiation (500 R) on the cell cycle of the diploid (EAT_dipl.) and the hypertetraploid (ELT) Ehrlich ascites tumor growing in the peritoneal cavity of male NMRI mice has been studied autoradio-graphically by the double labelling technique (H³- and C¹⁴-thymidine) and the method of labelled mitoses. The results suggest that the proliferation kinetics of tumor cells after irradiation depends on the type of tumor cell strain tested. Identical results were obtained after application of the alkylating agent cyclophosphamide (100 mg/kg).     

Wound-induced alterations in survival of60Co irradiated mice: importance of wound timing

Summary Wounding mice shortly before or shortly after lethal⁶⁰Co irradiation enhances survival. Survival of wounded-irradiated mice may be due to enhanced hematopoietic recovery as measured by endogenous spleen colony (E-CFU-s) formation.Supported by the Armed Forces Radiobiology Research Institute, Defense Nuclear Agency, under research Work Unit 00129. Views presented in this paper are those of the authors; no endorsement by the Defense Nuclear Agency has been given or should be inferred.Research was conducted according to the principles enunciated in the Guide for the Care and Use of Laboratory Animals' prepared by the Institute of Laboratory Research, National Research Council.     

Photoperiodic induction of diapause in an insect is vitamin A dependent

Summary A photoperiodic response was found to be absent in larvae of the parasitoid waspApanteles glomeratus when its host (caterpillars ofPieris brassicae) was reared on a low-carotenoid artificial diet. Addition of vitamin A to the host's diet restored the response to short-day photoperiods in the wasp larvae, thus showing that vitamin A is essential for photoperiodic induction of diapause. Possibly vitamin A or a derivative of vitamin A functions as the photoreceptor pigment for the photoperiodic reaction in this species of insect.     

Acute60Co-gamma irradiation of rats decreases the inhibitory effect of succinate on the lipid peroxidation of liver mitochondria

Summary Succinate inhibits NADPH-dependent lipid peroxidation of liver mitochondria. This effect of succinate decreased 12 h after whole-body⁶⁰Co-gamma irradiation, depending on the dose of irradiation.     

De l'emploi d'isotopes radioactifs artificiels,dans le but d'exercer un effet radio-biologique localisé

Summary One of the authors has previously reported on a method which consists in the utilization of an artifical radioactive isotope (Zn⁶³), suspended in a suitably prepared solution ofpectin, for the production oflocalized biological radiation effects.This « macromolecular occlusion » of the radioactive isotope enables one to perform intraperitoneal injections (in cases of cancer of the ovaries with severe metastatic peritoneal extension), evidently also instillations in cavernous organs, and furthermore direct intratumoral injections, without diffusion of the radioactivity outside the treated areas, as shown both by autoradiographs and controls of blood and urine specimens with a Geiger counter.The authors investigated further whether this procedure would also be suitable for obtaining, by means ofintravenous injections, alocalized radiation effect within thelungs, as presumably the radiozinc, held in the large molecules of pectin, could thus be retained in the pulmonary capillaries. Intravenous injections of such a pectin solution containing radiozinc were performed on rabbits, and autoradiographic controls gave evidence of this expected fixation within the lungs.For the purpose of preliminary clinical investigation 40 millicuries of Zn⁶³ suspended in 6 cm³ of a 3 p. c. isotonic pectin solution were injectedintravenously in a female patient with mainly pulmonary metastases of a previously operated hypernephroma. This patient had been also submitted to X-ray therapy. In spite of a poor general condition, the injection was well tolerated. Autoradiographic controls showed quite clearly that the radioactivity remains precisely localized within the pulmonary areas. No radioactivity whatsoever was demonstrated with the counter in the urine eliminated by this patient after the injection, a fact which points to a rather amazing accuracy of the fixation of the radiozinc in the lungs. This first clinical experience seems quite interesting in view of improving the therapeutic possibilities of pathological, especially neoplastic pulmonary conditions.     

Some effects of radiation on the free amino acids of adult female Mediterranean fruit fly,Ceratitis capitata Wied

Summary Ceratitis capitata pupae, 2–3 days before adult emergence, were treated with gamma irradiation from a⁶⁰Co source. The female fruit flies were extracted and analyzed for free amino acids.     

Ethanol as a ‘food’ forDrosophila melanogaster: Influence of theEbony gene

Summary The survival time of adultDrosophila melanogaster flies without food is greater in the presence of ethanol, especially for flies of strains or lines with a higher alcohol dehydrogenase activity. It seems that theebony gene can act in some populations as a selective factor favoring the ADH^F allele, as well as the minor genes enhancing the alcohol dehydrogenase activity level.Chargé de Recherches au F. N. R. S.     

The impact of cyclin-dependent kinase 5 depletion on poly(ADP-ribose) polymerase activity and responses to radiation

Cyclin-dependent kinase 5 (Cdk5) has been identified as a determinant of sensitivity to poly(ADP-ribose) polymerase (PARP) inhibitors. Here, the consequences of its depletion on cell survival, PARP activity, the recruitment of base excision repair (BER) proteins to DNA damage sites, and overall DNA single-strand break (SSB) repair were investigated using isogenic HeLa stably depleted (KD) and Control cell lines. Synthetic lethality achieved by disrupting PARP activity in Cdk5-deficient cells was confirmed, and the Cdk5^KD cells were also found to be sensitive to the killing effects of ionizing radiation (IR) but not methyl methanesulfonate or neocarzinostatin. The recruitment profiles of GFP-PARP-1 and XRCC1-YFP to sites of micro-irradiated Cdk5^KD cells were slower and reached lower maximum values, while the profile of GFP-PCNA recruitment was faster and attained higher maximum values compared to Control cells. Higher basal, IR, and hydrogen peroxide-induced polymer levels were observed in Cdk5^KD compared to Control cells. Recruitment of GFP-PARP-1 in which serines 782, 785, and 786, potential Cdk5 phosphorylation targets, were mutated to alanines in micro-irradiated Control cells was also reduced. We hypothesize that Cdk5-dependent PARP-1 phosphorylation on one or more of these serines results in an attenuation of its ribosylating activity facilitating persistence at DNA damage sites. Despite these deficiencies, Cdk5^KD cells are able to effectively repair SSBs probably via the long patch BER pathway, suggesting that the enhanced radiation sensitivity of Cdk5^KD cells is due to a role of Cdk5 in other pathways or the altered polymer levels.     

Increase of proteolytic activity inLactobacillus casei by X-ray irradiation

Summary A study was conducted on the effect of X-ray irradiation on proteolytic activity ofLactobacillus casei (RTS). Substantial increase (82%) in the proteolytic activity was noted among the isolates of X-ray irradiation survivors after 18,000 R.     

Local action of vitamin A on amphibian limb regeneration

Summary Axolotl forearms isolated from upper arm tissue either surgically or by local irradiation regenerate excessive structures after vitamin A treatment. This demonstration excludes the possibility of regeneration being altered by enhanced cellular migration and thus indicates that vitamin A respecifies cells close to the site of amputation.     

Vitamin E,catalase, manganous or cobaltous ions and dithiothreitol protect against Tween 20-induced hemolysis of vitamin E-deficient chick and kid erythrocytes

Summary The effective Tween 20 concentration at which 70% hemolysis was achieved in vitro correlated with the plasma vitamin E content of chicks (r=0.85). Addition of catalase, MnCl₂, CoCl₂ or dithiothreitol in vitro showed significant protection against the hemolysis induced by Tween 20 in vitamin E-deficient chick and kid erythrocytes.