Performance of 133 compounds in the lambda prophage induction endpoint of the Microscreen assay and a comparison with S. typhimurium mutagenicity and rodent carcinogenicity assays.

The Microscreen assay was developed as a means of testing very small samples, as in complex mixture fractionation. It is a multi-endpoint assay which utilizes E. coli WP2s(lambda). Exposure takes place to serial dilutions of the test compound in microtitre wells (250 microliters) followed by sampling from wells in which growth has occurred ('non-toxic wells'). Although a number of different endpoints can be measured, only the prophage induction endpoint (the first one developed) has been extensively tested. Results with 133 compounds are presented. These include 111 compounds which have been tested in the S. typhimurium assay and 66 compounds for which both rodent bioassay and S. typhimurium assay data exists. The concordance for the Microscreen assay and the S. typhimurium assay was 71%. For this group of compounds, the sensitivity of the Microscreen assay in detecting carcinogens was 76% compared with 58% for the S. typhimurium assay. However, the S. typhimurium assay was somewhat more specific (69%) compared with the Microscreen (56%). The overall association between carcinogenicity and Microscreen results was statistically significant (p = 0.029), whereas for the S. typhimurium assay the association with carcinogenicity was non-significant (p = 0.086). The Microscreen assay was able to detect halogenated compounds better than the S. typhimurium assay. The Microscreen assay should prove useful in complex mixture fractionation, or in other situations where sample size is limiting.     

Production and response to interleukin 2 in vitro and in vivo after bone marrow transplantation in mice

Lymphoid cells from bone marrow radiation chimeras do not produce normal levels of IL 2 but are capable of responding to IL 2 in mitogenic and cytotoxic assays in vitro. The administration of recombinant human IL 2 into host mice that have received allogeneic, H-2-compatible marrow did not enhance mortality.     

Analysis of recombinant DNA clones of the endogenous BALB/c murine leukemia virus WN1802N: variation in long terminal repeat length

We analyzed 15 recombinant DNA clones of the unintegrated closed circular DNA intermediate of the BALB/c endogenous ecotropic murine leukemia virus WN1802N. Thirteen of these clones had an insert which corresponded to the complete murine leukemia virus genome. Of these, six contained a single long terminal repeat (LTR) and seven contained two LTRs. The viral genomes in nine clones had an LTR of 520 base pairs (bp), one had an LTR of 570 bp, three had an LTR of 600 bp, and one had an LTR of 670 bp. Restriction endonuclease analysis demonstrated that the size variability resides in the U3 region. Seven of eight clones which yielded infectious virus by DNA transfection had the 520-bp LTR, and the other had a 600-bp LTR. More detailed examination of plasmid subclones of three isolates with different-sized LTRs revealed that the approximate position which varies in the U3 region corresponds to the 72-bp repeat region of Moloney sarcoma virus. Possible consequences of these variations are discussed.     

Reversal of Fv-1 host range by in vitro restriction endonuclease fragment exchange between molecular clones of N-tropic and B-tropic murine leukemia virus genomes.

We molecularly cloned unintegrated viral DNA of the BALB/c endogenous N-tropic and B-tropic murine leukemia retroviruses and in vitro passaged N-tropic Gross (passage A) murine leukemia retroviruses. Recombinant genomes were constructed in vitro by exchanging homologous restriction enzyme fragments from N- or B-tropic parents and subsequent recloning. Infectious virus was recovered after transfection of these recombinant genomes into NIH-3T3 cells and cocultivation with the Fv-1 nonrestrictive SC-1 cells. XC plaque assays of recombinant virus progeny on Fv-ln and Fv-lb cells indicated that the Fv-l host range was determined by sequences located between the BamHI site in the p30 region of the gag gene (1.6 kilobase pairs from the left end of the map) and the HindIII site located in the pol gene (2.9 kilobase pairs from the left end of the map).     

L'hématoxyline au plomb permet-elle l'identification des cellules corticotropes de l'hypophyse des téléostéens?

Conclusion La technique à l'hématoxyline au plomb de Mac Conaill permet l'identification d'un type cellulaire situé dans la pars distalis rostrale, en bordure des ramifications de la neurohypophyse dans l'hypophyse de plusieurs Poissons Téléostéens; il est également colorable par le bleu d'alizarine d'Herlant; toutefois, l'intensité de ces deux réactions tinctoriales est très variable selon les espèces. Chez l'Anguille, les Salmonidés et le Cyprin, la nature corticotrope de ces cellules est établie expérimentalement. La colorabilité par l'hématoxyline d'une autre catégorie cellulaire dans la pars intermedia laisse supposer l'existence de constituants communs aux deux types cellulaires, et peut-être même à leurs produits d'élaboration. Cette communauté de certains aminoacides entre l'ACTH et l'intermédine a déjà été démontrée chez divers Mammifères.     

Réactions observées chez l'Anguille maintenue dans un milieu privé d'électrolytes,en particulier au niveau du système hypothalamo-hypophysaire

Résumé 45 Anguilles sont maintenues (32 h. à 46 jours) en eau déminéralisée (désionisée ou distillée). On note un assombrissement de la pigmentation, une hypertrophie importante de la vésicule biliaire et des modifications du système hypothalamo-hypophysaire.Dans la pars distalis rostrale, les cellules à prolactine subissent une régression, ne semblant plus élaborer de granulations. Les cellules corticotropes sont faiblement stimulées, partiellement dégranulées, en accord avec la légère activation de l'interrénal. Les cellules thyréotropes apparaissent souvent peu actives après 46 jours; cependant, l'examen préliminaire des thyroïdes ne montre pas d'hypoactivité. Dans la pars distalis proximale, les cellules gonadotropes ne sont pas modifiées; les cellules somatotropes sont stimulées: hypertrophie nucléaire et nucléolaire, développement de l'ergastoplasme, sans dégranulation complète. Leur rôle dans l'osmorégulation est discuté. Dans le lobe intermédiaire, les cellules du type 1, hématoxyline au plomb positives, prédominent chez les témoins; elles subissent une stimulation avec dégranulation partielle correspondant peut-être à l'assombrissement de la pigmentation. Le type 2, PAS positif, s'hypertrophie et s'hyperplasie rapidement en eau déminéralisée. Cette hyperactivité indiquerait un rôle dans l'osmorégulation. Le neuroséorétat s'accumule, puis tend à diminuer, la neurohypophyse étant de volume réduit. Le noyau préoptique paraît peu actif. L'hypothèse d'une faible utilisation des produits élaborés par l'axe hypothalamo-neurohypophysaire est envisagée. En plus de l'interrénal et des corpuscules de Stannius, l'hypothalamus et l'hypophyse interviennent donc dans la réponse de l'Anguille à une déminéralisation du milieu ambiant.

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Summary 45 male silver eels were kept from 32 hours to 46 days in demineralized (distilled or deionized) water. They show a darkening of the skin, a hypertrophy of the gall-bladder and some modifications of the hypothalamo-neuro-adenohypophysial system.In the rostral pars distalis, the prolactin cells regressed and appeared chromophobic. The corticotrophic cells are slightly stimulated and partly degranulated, in correlation with some activation of the interrenal, previously described. The thyrotrophic cells do not appear active at the end of the experiment, however the thyroid glands are not often inactive.In the proximal pars distalis, the gonadotrophic cells remain undifferentiated. The somatotrophic cells are strongly stimulated: a cellular, nuclear and nucleolar hyperactivity, without degranulation and a development of the ergastoplasm are observed. Their role in osmoregulation and metabolism of potassium is discussed.The pars intermedia is composed of two cell types: one of which is predominant, leadhematoxylin positive. It could elaborate the MSH (intermedin). In demineralized water, it is partly degranulated and progressively stimulated, perhaps in correlation with the darkening of the pigmentation. The other cell type is PAS positive, and not abundant in the control fish. In the treated fish, rapid hyperplasia and hypertrophy occur, with cytoplasmic degranulation and mitotic activity, so that it becomes predominant in this lobe after a month or more in demineralized water. Cytologic criteria indicate a great hyperactivity which seems to play a role in the processes of the osmoregulation; this hypothesis is discussed.The neurosecretory material tends to increase, as a storage, then to decrease in the neurohypophysis which is of a reduced volume. The preoptic nucleus does not appear active. In addition to the modifications of the interrenal and the corpuscles of Stannius previously described, the changes in the hypothalamus and the pituitary constitute evidence of their important role in the reaction of the eel to a demineralized environment.

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Ce travail a été réalisé avec la collaboration technique de Mademoiselle Jacqueline Olivereau, du C.N.R.S., que nous remercions bien vivement: elle nous a beaucoup aidée au cours de ces expériences, en particulier lors des changements quotidiens de l'eau distillée ou désionisée, et a réalisé toutes les préparations histologiques et les microphotographies illustrant cet article.