[Does nicotine add to the carcinogenic strain of tobacco smoke?]. / Trägt Nikotin zur Krebsentstehung im oberen Aerodigestivtrakt bei?

BACKGROUND: It is accepted that nicotine in tobacco smoke causes addiction via nicotinic acetylcholine receptors in the central nervous system. For a long time, the tumorigenic potential of smoking was attributed to compounds other than nicotine. However, more recently data have accumulated which suggest that nicotine may add to the cancer risk by stimulating cellular growth via non-neuronal acetylcholine receptors, by suppressing apoptosis, and by inducing angiogenesis not only in atheromatous plaques but also in tumors. In the present study the possible direct genotoxic effects of nicotine on DNA were investigated in human target cells of carcinogenesis in the upper aerodigestive tract. PATIENTS AND METHODS: Human nasal mucosa, lymphatic tissue of the palatine tonsils, supraglottic epithelium of the larynx, and peripheral lymphocytes were exposed to rising concentrations of nicotine. DNA damage was investigated by alkaline single-cell microgel electrophoresis (Comet) assay. Cytotoxicity was assessed by trypan blue exclusion. RESULTS: Nicotine induced dose-dependent DNA damage in all cell types at low cytotoxic concentrations that allowed viabilities well above 80%. The lowest nicotine concentrations eliciting a significant increase in DNA migration were 1 mM for tonsillar cells and 0.25 mM for all other cell types. CONCLUSION: Nicotine induces genotoxic effects in human target cells of carcinogenesis in the upper aerodigestive tract at relevant concentrations. Thus, nicotine may contribute directly to tumor initiation resulting from smoking.

[Mini-organ cultures of human nasal mucosa. A model for eco-genotoxicological investigations]. / Miniorgankulturen humaner nasaler Mukosa. Ein Modell für ökogenotoxikologische Untersuchungen.

BACKGROUND: Volatile and ingestive xenobiotics may induce cancer in the mucosa of the upper aerodigestive tract. A new model is presented combining mini-organ cultures of human mucosa and the Comet assay that allows investigation of tumor initiation steps in vitro. METHOD: Specimens of human mucosa of the inferior nasal turbinates were cultured as mini-organs and exposed to xenobiotics once, twice or three times with consecutive repair intervals. The cultures were monitored for structural integrity (inverse microscopy, histology), DNA fragmentation and repair activity (Comet assay), induction of apoptosis (annexin V assay), and production of IL-8 and GM-CSF (ELISA). RESULTS: Mini-organ cultures showed a good structural integrity during the whole culture period. Exposure to N-nitrosodiethylamine (NDEA) and benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE) induced significant DNA fragmentation. Sodium dichromate (Na2Cr2O7) had an additive DNA fragmentation effect with repetitive exposure. Significant DNA repair was seen after strand break induction by Na2Cr2O7, only. Apoptosis was seen after three exposures to BPDE und Na2Cr2O7, but not NDEA. Inflammatory cytokine release was unaltered by NDEA. However, BPDE and Na2Cr2O7 reduced GM-CSF and Na2Cr2O7 reduced IL-8 excretion. CONCLUSION: This three dimensional mini-organ culture system proved to be very helpful in characterizing volatile and ingestive xenobiotics potentially hazardous to humans. Beside the information concerning genotoxicity, it allows cytological and immunological studies. In contrast to investigations with fresh specimens, repetitive or chronic exposure to xenobiotics is possible in mucosal cells with their epithelial structural integrity. Therefore, mini-organ cultures of human upper aerodigestive tract epithelia represent a model closely resembling the in vivo situation.

Toxicological assessment of noxious inhalants.

In the past centuries mankind has been exposed to various forms of air pollution not only at his occupational but also in his social environment. He mainly gets exposed with these pollutants through the respiratory organs and partially absorbs them into the body. Many of these airborne substances can be harmful for humans and some of them may account for tumorigenic effects.The following essay describes the main features of toxicological assessment of inhalative environmental and workplace xenobiotics. The essay also explains relevant characteristics and limit values of noxious compounds and gases and depicts modern testing methods. To this end, emphasis is given on methods characterizing the different stages of tumorigenic processes. Various test systems have been developed which can be used in vivo, ex vivo or in vitro. They are to a great part based on the evidence of changes in DNA or particular genes of cells. Among others they have highlighted the impact of interindividual variability on enzymatic activation of xenobiotics and on susceptibility of the host to tumor diseases.Unfortunately, for many inhalative environmental noxious agents no sufficient risk profiles have been developed. The completion of these profiles should be the goal of toxicological assessment in order to allow reasonable socioeconomic or individual-based risk reduction.

Effects of naloxone infusion on plasma levels of LH, FSH, and in addition TSH and prolactin in males, before and after oestrogen or anti-oestrogen treatment.

The inhibitory action of endogenous opioids on gonadotrophin release is now well documented. Since LHRH-producing neurons do not possess oestrogen-receptors, it is likely that some other compound mediates the negative feedback action of oestrogens on the gonadotrophin release in the male. To test the hypothesis that endogenous opioids are implicated in this negative feedback action in the human male, the opioid receptor antagonist naloxone (2 mg/h for 4 h) was infused into 7 normogonadotrophic oligozoospermic men before and after 6 weeks of treatment with the oestrogen-receptor antagonist tamoxifen (TAM) (10 mg twice daily) and 6 eugonadal transsexual males before and after 6 weeks of administration of ethinyloestradiol (EE) (10 micrograms three times a day). The effects of naloxone on TSH and prolactin (PRL) release were also studied. Naloxone administration resulted in a significant release of gonadotrophins, but not of TSH and PRL. Administration of oestrogen and anti-oestrogen did not significantly affect the response of gonadotrophins to naloxone infusion and no evidence of consistently antagonistic effects of oestrogen and anti-oestrogen on the naloxone-induced gonadotrophin release was obtained. This shows that endogenous opioids are probably not intermediary in the negative feedback control of oestrogens on gonadotrophin release in the human male. Surprisingly, in contrast to the eugonadal transsexual males, FSH levels in the oligozoospermic men did not respond to naloxone administration. As naloxone is thought to exert its action on gonadotrophin release via a disinhibition of endogenous LHRH release, this finding is unexpected. Exogenous LHRH administration leads to a normal response of FSH in normogonadotrophic oligozoospermic men. No plausible explanation for this finding can presently be offered.

Effects of continuous LHRH infusion on plasma levels of LH and FSH in males, before and after oestrogen or anti-oestrogen treatment.

In order to study the role of oestrogens on gonadotrophin release in the human male, LHRH was administered as an infusion at a constant rate of 0.5 micrograms/minute for 4 hours to 7 normogonadotrophic oligozoospermic men, 6 eugonadal male-to-female transsexuals and 9 eugonadal male volunteers. In agreement with in vitro data a biphasic release pattern of both LH and FSH was observed in eugonadal transsexuals as well as in normogonadotrophic oligozoospermic men. In the latter the release of LH was greater than in eugonadal transsexual males and volunteers, which points to a different functioning of the hypothalamic-pituitary unit in normogonadotrophic oligozoospermic men. On the other hand the FSH response to LHRH stimulation was normal in these men. Three months' treatment with the oestrogen-receptor antagonist tamoxifen (TAM) (10 mg twice daily) in the normogonadotrophic oligozoospermic men stimulated basal LH, FSH and testosterone (T) levels. The fact that gonadotrophin levels rose in spite of increased T levels, suggests a role of endogenous oestrogens in the negative feedback regulation of gonadotrophin release in these men. Upon TAM treatment the first phase, the plateau and the second phase of LH release were augmented, whereas only the plateau and the second phase of FSH release were increased. Six weeks' administration of the oestrogen ethinyloestradiol (EE) (10 micrograms three times a day) in the eugonadal transsexual males suppressed basal T and oestradiol (E2) levels without affecting basal gonadotrophin levels significantly. In EE-treated males the first phase of LH release tended to be lower, whereas the plateau of LH had decreased significantly. The second phase of LH was unaffected.(ABSTRACT TRUNCATED AT 250 WORDS)

[Biology, serology and biochemistry of leucemogenic viruses derived from a radiation-induced tumor of C57BL mice]. / Biologie, sérologie et biochimie de virus leucémogènes dérivés d'une tumeur radioinduite de la Souris C57BL.

Two distinct rat propagates of a radiation leukemia virus (RadLV-Rs) from the C57BL mouse respectively induced characteristic leukemogenic effects. These were found to be related with the infection titers of the isolates, but not with either their antigenic specificities or their viral and proviral genome sequences.

Detection and spontaneous alteration of lymphocyte antigens on slide smears.

Smears of cell suspensions from murine lymphoid organs were prepared on slides, air dried, and processed for detection of immunoglobulin (Ig) and theta (Thy-1) antigens by the unlabeled antibody peroxidase-antiperoxidase (PAP) technique. Satisfactory results were obtained for both antigens on recently made smears. However, slides kept at room temperature showed a progressive decrease in staining of the two antigens with time. Various fixatives and preservation procedures were tested to prevent this alteration. Good conservation of smears was obtained when slides were kept at -18 degrees C and/or air isolated before or after fixation with alcohols. A similar degradation of Ig and/or Thy-1 antigens occurred also in histological tissue sections or in serum spots dried on slides. The major cause for this degradation is thought to be contact with air, residual enzymatic hydrolysis playing a less important role.

Radioimmunoassay determination of two main cross-reacting viral antigens during radiation and Rauscher induced leukemias.

The two main proteins p 30 and gp 70 from the Rauscher leukemia virus (RLV) have been isolated, labeled with 125I and used in radioimmune competition assays to estimate the amount of cross-reacting antigens produced during the evolution of various leukemias. The Rauscher leukemia in Balb/c mice, the radiation induced leukemia (RadLV) in C57Bl mice and two types derived therefrom by serial passages in mice (RadLV-RS) and in rats (RadLV-rat) were studied. Whereas the p 30 from RLV or RadLV (rat) viruses showed complete identity, the cross-reaction of their gp 70 proteins wasonly partial. The main findings in the tissues were the following: 1. The concentration of p 30 and gp 70 antigens increased much more during the RadLV-rat leukemia than in irradiated or RadLV (RS) treated mice. 2. In the serum, the ratio p 30/gp 70 was in general higher than one in the Rauscher leukemic mice and less than one in all types of RadLV leukemias. 3. In the spleen, RLV and RadLV (RS) infected mice had higher levels of p 30 than of gp 70. The reverse occurred in irradiated mice and in leukemic rats. Finally, the catabolic degradation of labeled RLV gp 70 was similar in normal and leukemic mice.

Specific precipitation of mouse and rat alpha 2-macroglobulin by immune sera directed against murine leukemia viruses.

Leukemia viruses preparations obtained from plasma of leukemic mice or rats were used to immunize rabbits. By immunoelectrophoresis, these non-absorbed antisera reacted specifically with one single mouse serum alpha 2 globulin which was further identified as alpha 2-macroglobulin by several criteria, including gel chromatography and nitracentrifugation. When mouse plasma derived virions were used as antigen, they gave rise to antibodies cross-reacting completely with the corresponding mouse and guinea pig protein and partially with rat and calf globulin. Immunization of rabbits with rat plasma derived virions induced antibodies directed preferentially against the rat alpha 2-macroglobulin.

Regulation of the neutral phosphatase in Chlamydomonas reinhardi: an immunogenetic study of wild-type and mutant strains.

In Chlamydomonas reinhardi, the activity of the neutral phosphatase considerably increases when the cells are grown in the absence of inorganic phosphate (Pi). A comparative immunological study of cells grown on media containing Pi or not indicated that the neutral phosphatase was synthesized de novo. Ten mutants lacking the neutral phosphatase and distributed among three genetic loci (PD2, PD3, PD24) were investigated for their ability to produce cross-reacting material (CRM) antigenically related to the wild enzyme. All mutants were shown to form much less CRM than the wild-type strain. It is proposed that the three genes are involved in the regulation of neutral phosphatase synthesis.

[Genetic control in the mouse]. / Le contrôle génétique chez la souris

Spontaneous mutations and environmental variations are responsible for the biological instability of inbred strains of mice. As far as possible, all external factors should be kept constant in a modern animal house. Several techniques to analyse part of the genome are available. Due to this limitation, it would be useful to have several laboratories participating to the genetic control.