A circular channel crucible oscillating viscometer: Detection of DNA damage induced in vivo by exceedingly small doses of dimethylnitrosamine

A new oscillating crucible viscometer, having a U-shaped circular channel, is described. The damping coefficient δ is lowered by an increase of the viscosity η. The instrument described here allows the solution to come in contact with inert plastic only. At all steps of its preparation and during viscosity measurements, giant DNA from rat liver nuclei was maintained at shear stresses around 10^?4 dynes cm^?2. Viscosity was studied as a function of surface tension, DNA concentration and shear stress. It was found that under our experimental conditions it was possible to obtain meaningful values for reduced viscosity, η_red, practically identical to intrinsic viscosity [η]. Rat liver nuclei are incubated in an alkaline lysing solution (pH 12.5; 22 °C): they are lysed immediately and the released DNA starts to uncoil. The viscosity of solutions of this giant DNA increases very slowly with time, reaching a maximum only after about ten hours. The process was accelerated by single-stranded breaks arising from methylation of DNA in vivo with dimethylnitrosamine. It was found that the time of DNA disentanglement was sensitive to an exceedingly small number of breaks. We think that we were able to measure molecular weights around the length of the single strand of an average chromosome (M_n 5 × 10¹⁰). An empirical relation between molecular weight and reduced viscosity after complete disentanglement was also established, as a linear log-log plot, covering a molecular weight range between 10⁸ and 2.5 × 10¹⁰. It is suggested that the viscosimetric evaluation of DNA disentanglement is probably the most sensitive method for studying DNA damage induced “in vivo” by chemical carcinogens.     

Exact versus heuristic models of kin selection

This paper examines the conditions under which the classical inclusive fitness formulation of Hamilton (1964) provides an adequate approximation to the dynamics of gene frequency change and to conditions for genetic equilibrium, in the “additive” model of altruism between sibs of Uyenoyama and Feldman (1981). It is concluded that the classical formulation is adequate, provided that either the effect of the gene on the probability of behaving altruistically is low or the costs and benefits of altruism are small, unless the benefit/cost ratio k is very close to 2, the value that must be exceeded for altruism to be favoured. In addition, the gene for altruism must be underdominant, recessive or partially recessive in its effect on the probability of behaving altruistically, for the inclusive fitness predictions to break down significantly.     

Evaluation of proficiency testing as a method of assessing competence to screen cervical smears

Evaluation of proficiency testing as a method of assessing competence to screen cervical smears Regular proficiency testing of pathologists and cytotechnologists who undertake the analysis of cervical smears has been carried out in cytology laboratories in the North Thames (West) Region since 1989. The protocol followed is one that has been adopted nationally. Since the scheme started, laboratory personnel from 17 cytology laboratories in the Region have participated and seven rounds of testing have been completed. Nine hundred and seventy-one tests were carried out and a pass rate of 96.4% was recorded. Two hundred and forty-seven cytologists took the test on at least one occasion and 63 cytologists took part in all seven rounds. Our results indicate that proficiency testing is capable of detecting cytologists who consistently perform below an acceptable standard and require retraining. They also show that even the most competent screeners can miss an abnormal smear. Seven cytologists who had proved their competence to screen on six occasions, missed an abnormal smear on the seventh, despite the fact that they were screening under test conditions when their vigilance should have been at its peak. Our findings indicate that false-negative reporting will inevitably occur during manual screening, and emphasize the need for further research into the causes and prevention of screening error. Evaluation des tests d'aptitude comme méthode d'appréciation de la compétence à lire les frottis cervico-utérins de dépistage Le passage régulier de tests d'aptitude pour les pathologistes et les cytotechniciens qui prennent en charge l'analyse de frottis cervico-utérins est utilisé dans les laboratoires de cytologie de la région North Thames (West) depuis 1989. Le protocole choisi est l'un de ceux qui ont été adoptés au niveau national. Depuis le début de ce programme, 7 sessions de tests ont été organisées et le personnel de 17 laboratoires de cytologie de la région a participé. Neuf cent soixante et onze tests ont été pratiqués et le taux de participation enregistré est de 96,4%. Deux cent quarante sept cytologistes ont fait le test au moins une fois et 63 cytologistes ont passé les 7 sessions de tests. Nos résultats indiquent que les tests d'apitude sont capables d'identifier les cytologistes qui ont besoin d'une formation car leur résultats aux tests sont régulièrement en dessous des standards acceptables. Ils montrent également que même les “screeners” les plus compétents peuvent passer à côté d'un frottis anormal. Sept cytologistes ayant montré leur compétence pour la lecture des frottis de dépistage sur 6 tests n'ont pas été capable de détecter un frottis anormal sur le septième test, bien que cette lecture ait été faite dans des conditions de test où leur vigilance aurait dû être à son maximum. Ces résultats indiquent qu'il est impossible d‘éviter les faux négatifs au cours du “screening manuel” et confirment la n?essité de poursuivre les recherches sur les causes des erreurs de lecture et sur leur prévention. Die Ergebnisse der Laistungstests als Grundlage für die Fähigkeit gynäkologische Abstriche auszuwerten Regelmässige Leistungstests von Zytopathologen und Zytotechnikerinnen beim Auswerten gynäkologischer Abstriche wurden seit 1989 in den Labors der Nord-Themse Region durchgeführt. Das Testprotokoll entspricht dem landesweit angewandten. Inzwischen nahmen 17 Labors an insgesamt 7 Testrunden teil. Die Gesamtzahl der Tests beträgt 971 wobei 96,4% der Teilnehmer die Tests bestanden. 247 Teilnehmer nahmen an zumindest einem Test, 63 an allen sieben teil. Die Auswertungen ergaben, dass auf diese Weise Zytologen mit unterdurch schnittlicher Leistung identifiziert und einem Zusatztraining zugeführt werden können. Sie zeigen aber auch, dass selbst die Erfahrendsten gelegentlich positive Befunde übersehen. Sieben Teilnehmer begingen diesen Fehler nach sechs fehlerfreien Durchgängen beim siebten Test trotz der bekannten Testsituation. Dies bedeutet, dass Fehler bei nichtautomatischer Auswertung unvermeidbar sind. Die Ursachenforschung muss weitergeführt werden.     

Synthèse et étude R.M.N. de disaccharides et trisaccharides dans la série du l-rhamnose

Various di- and tri-saccharides containing l-rhamnose were synthesized by condensation of 2,3,4-tri-O-acetyl- or 2,3,4-tri-O-benzoyl-α-l-rhamnopyranosyl bromide with an unblocked glycopyranoside. The determination of the anomeric configuration of l-rhamnose saccharides by n.m.r. is difficult because structure has a greater effect on the spectra than does configuration. The α and β configurations and the position of the substitution may be assigned from the chemical shifts of H-5 and CH₃. In all the compounds having a β configuration, a shielding of the methyl group and a deshielding of the H-5 proton have been observed as compared to the compounds having an α configuration. The H-5 proton and the methyl group of peracetylated, (1→3)-linked α-l derivatives always resonate at higher fields than the corresponding protons of (1→6)-linked α-l derivatives.     

Seed bank dynamics and tree‐fall gaps in a northwestern Mexican Quercus‐Pinus forest

Questions: How does the seed bank respond to different types of tree‐fall gaps and seasonal variations? How does the soil seed bank influence recovery of the standing vegetation in the mature forest and tree‐fall gaps? Location: 1800 — 2020 m a.s.l., Quercus‐Pinus forest, Baja California Sur, Mexico. Methods: Seed size, species composition and germination were estimated under different environmental conditions during dry and rainy seasons: a mature forest plot and gaps created by dead standing trees, snapped‐of f trees and uprooted trees. The soil seed bank was investigated using direct propagule emergence under laboratory conditions, from soil cores obtained during both seasons. Results: 21 species, 20 genera and 14 families constitute the seed bank of this forest community. Fabaceae, Asteraceae, Euphorbiaceae and Lamiaceae were the most frequently represented families in the seed bank. Floristic composition and species richness varied according to the different modes of tree death. Species composition of seed banks and standing vegetation had very low similarity coefficients and were statistically different. Seed bank sizes varied between 164 and 362 ind.m^‐2 in the mature forest plot for the dry and rainy seasons, respectively, while soil seed bank sizes for gaps ranged between 23–208 ind.m^‐2 forthe dry season and between 81–282 ind.m^‐2 for the rainy season. Conclusions: Seed bank sizes and germination response were always higher in the rainy season under all the environmental conditions analysed. Results suggest that timing responses to gap formation of the soil seed bank could be more delayed in this temperate forest than expected.     

Deuterium stable isotope ratios as tracers of water resource use: an experimental test with rock doves

Naturally-occurring deuterium stable isotope ratios can potentially be used to trace water resource use by animals, but estimating the contribution of isotopically distinct water sources requires the accurate prediction of isotopic discrimination factors between water inputs and an animals body water pool. We examined the feasibility of using estimates of water fluxes between a bird and its environment with a mass-balance model for the deuterium stable isotope ratio of avian body water (D_body) to predict isotopic discrimination factors. Apparent fractionation and thus discrimination factors were predicted to vary with the proportion of an animals total water losses than could be attributed to evaporative processes. To test our ability to predict isotopic discrimination, we manipulated water intake and evaporative water loss in rock doves (Columba livia) by providing them with fresh water or 0.15 M NaCl solution in thermoneutral or hot environments. After we switched the birds from drinking water with D=–95 VSMOW (Vienna Standard Mean Ocean Water) to enriched drinking water with D=+52 VSMOW, steady-state D_body was approached asymptotically. The equilibrium D_body was enriched by 10–50 relative to water inputs. After isotopic equilibrium was reached, the degree of enrichment was positively related (r²=0.34) to the fraction of total water loss that occurred by evaporation supporting the major prediction of the model. The variation we observed in discrimination factors suggests that the apparent fractionation of deuterium will be difficult to predict accurately under natural conditions. Our results show that accurate estimates of the contribution of different water sources to a birds body water pool require large deuterium isotopic differences between the sources.     

Reproductive phenology of a natural population of Armadillidium pelagicum Arcangeli, 1955 (Isopoda, Oniscidea)

Armadillidium pelagicum, the most abundant species of the genus in Tunisia is endemic to the North of Tunisia and the circumsicilian islands. In the laboratory and under the natural conditions of temperature and photoperiod, virgin females of A. pelagicum exhibit a spontaneous ovarian maturation, followed by a parturial moult. Nevertheless, the onset of reproduction is greatly accelerated by the presence of a male. In fact, mating shortened the lag time (from the beginning of experiment to the parturial moult) by reducing the number of normal moult preceding the first parturial moult and the duration of the preparturial intermoult. In this species, only one mating is enough to ensure the fertility for several egg layings. Otherwise, it has been shown that A. pelagicum has a reproductive period rather than a reproductive activity and that mating females have a longer reproductive period than virgins. Like several species of terrestrial Isopods, the duration of the gestation period is temperature-dependent.     

Apport de la TEP/TDM au 18FDG dans le bilan préopératoire des phéochromocytomes. Étude rétrospective comparative par rapport à la scintigraphie à la MIBG

Pheochromocytomas (PHEO) are tumors arisen from the adrenal medulla, with a high secretory risk. Malignancy is rare and difficult to establish before metastastic spread. The risk of multifocality becomes greater whether genetic predisposition exists. MIBG scintigraphy is the reference functional imaging of PHEO. The place of 18FDG PET/CT is not well-established in literature. Our study retrospectively analyzes patients operated for a PHEO and who underwent both MIBG and 18FDG PET/CT before surgery, between 2007 and 2015. On the 49 patients included (52 lesions), among them 13 had genetic mutation, MIBG detected 39 lesions (75%) and 18FDG PET/CT forty-eight (92%), enabling a combined sensitivity of 98%. Sensitivity was not affected by the predominant secretion (metanephrine or normetanephrine), whereas MIBG-negative lesions showed a higher proliferation index (Ki67) than MIBG-positive lesions (6.6 vs. 2.8; P = 0.0044). FDG PET/CT semi-quantitative indices vary with the germline mutation status and were significantly higher for Cluster 1 lesions (SDHx, VHL or FH mutations) than for any other lesions (SUVmax = 27.73 vs. 5.92 for the others mutations (Cluster 2), 9.53 for lesions without mutation and 3.78 for undetermined lesions; P = 0.002). In conclusion, because of their capacity to take up FDG, 18FDG PET/CT could be useful in the preoperative imaging of PHEO, especially when MIBG is not contributive or when F-DOPA PET/CT is not available. An intense FDG uptake may orient to a Cluster 1 mutation.