A rapid and sensitive method for the detection of histone peptides

Fluorescamine has been used to obtain a peptide map of a mixture of histones (H₃, H₂A, H₂B, and H₄) prepared from oocytes of Xenopus laevis. Fluorescamine was found to be more sensitive than o-phthalaldehyde or ninhydrin-Cd for the detection of peptide fragments obtained from tryptic digestion of oocyte histones of X. laevis and the peptic digestion of the β chain of insulin. Using the β chain of insulin for a comparison, the 8 major peptide fragments could be separated by electrophoresis within 30 min and were detectable at the picomols level. Some 70 peptide spots of X. laevis oocyte histones were resolved, thus permitting the analysis of this complex mixture of polypeptides without the need for prior separation.     

Reappraisal of the electron spin resonance spectra of maleimide and iodoacetamide spin labels in erythrocyte ghosts.

The effects of sulfhydryl inhibitors (iodoacetamide and N-ethylmaleimide) on the electron spin resonance spectra of two maleimide and two iodoacetamide spin labels in erythrocyte ghosts were found to correlate with their relative “lipid”/water partition coefficients. But the spectral characteristics of the maleimide spin labels, and their ghost concentrations after iodoacetamide inhibition, are not consistent with the hypothesis that interprets their spectra solely on the basis of a heterogenous membrane distribution. An alternative hypothesis is suggested which is compatible with relative “lipid solubilities” and the iodoacetamide inhibition spectra.     

A new method for determining sulfoxides in peptide molecules using x-ray photoelectron spectroscopy

The problem of the quantitative determination of sulfoxide groups in peptide molecules has been re-examined. The approaches currently available for the estimation of δ-sulfoxide amino acids are limited in number and characterized by serious shortcomings; in addition, the choice of methods for the estimation of γ-sulfoxide amino acids is even more restricted. A new, rapid, and nondestructive direct method for determining quantitatively all types of sulfoxides in peptide molecules by using x-ray photoelectron spectroscopy is described.     

Conservation and chromosomal localization of DNA satellites in balenopterid whales

DNA satellites were isolated from three balenopterid species, viz. the minke, sei, and fin whales. In each of them at least two DNA satellites were recognizable with buoyant densities in neutral CsCl of =1.702/1.703 and =1.710/1.711, respectively. cRNAs from each satellite group were used for filter and in situ hybridisations. Homo- and heterologous DNA-cRNA hybrids within each satellite group yielded virtually identical melting curve profiles showing conservation of at least a considerable part of the DNA satellite sequences. There was no evident sequence homology between the =1.702/1.703 and the =1.710/1.711 satellites by filter hybridisation. — The in situ hybridisation showed that in each species the =1.702/1.703 satellite was located in centromeric-paracentromeric C-bands in a few pairs, whereas the =1.710/1.711 satellite was located in terminal C-bands throughout the karyotypes. — The data on the whale DNA satellites indicate that the quantitative evolution of the satellite DNA sequences preceded species divergence of the balenopterids and that the satellite sequences have remained relatively unaltered since the divergence took place. The function of satellite DNA is considered to imply the introduction of both chromosomal and genic polymorphisms and thus being of great importance in speciation. Based upon these concepts a model is postulated for the function of satellite DNA. According to this model at meiotic pairing euchromatin-heterochromatin overlapping between homologous chromosomes is considered to be of a general occurrence. This overlapping is presumed to be accentuated by the size heteromorphism frequently observed between homologous heterochromatic segments (C-bands). In the region of such euchromatin-heterochromatin overlapping, crossing-over would be excluded. The overlapping is suggested to be rectified progressively in the chromosome arms, leaving unaffected crossing-over distant to the euchromatin-heterochromatin junctions. The consequence of this will be that genes in the proximity of the junctions are collectively inherited and selected, whereas genes distant to the heterochromatin will be independently assorted and selected.     

Phenotypic conversion of cultured mouse embryo cells by aza pyrimidine nucleosides.

Cells of the $\text{C3H}\text{10}\text{T}\text{1}\text{2}\text{CL8}$ line, which are nonmyoblastic in nature, form functional myotubes when treated with low concentrations of 5-azacytidine. Further characterization of the myotubes revealed that they arise from the fusion of mononucleated precursors and not as a result of endoreplication. They accumulate histochemically detectable myosin ATPase activity as well as acetylcholine receptors capable of binding radioactively labeled α-bungarotoxin. The deoxy analog, 5-aza-2′-deoxycytidine, induced myogenic conversion at one-tenth of the maximally effective concentration of 5-azacytidine. The ability of both analogs to induce myotube formation and to cause cytotoxicity was strongly influenced by cotreatment with certain pyrimidine nucleosides. These effects were consistent with a requirement for metabolism of both aza compounds to phosphorylated derivatives and with a mechanism of action based on their incorporation into DNA. Concentrations of the analogs causing myogenic conversion did not substantially alter rates of DNA, RNA, or protein synthesis as measured by precursor incorporation into intact cells. The induction of myotubes by 5-azacytidine in cells synchronized by two different methods required that treatment with the analog was carried out at a critical phase early in S phase. Thus the mechanism of drug action appears to be linked to specific DNA synthesis.     

Development of wheat bulb fly (Della coarctata fall.) larvae and pupae at different temperatures

When eggs of wheat bulb fly were added to young winter wheat plants in pots and kept at 5°, 8°, 10°, 12°, 15° and 20°, only 14%–44% of the eggs added produced larvae that succeeded in invading plants, but when newly hatched larvae were added directly 54%–92% were successful. At temperatures from 5° to 25°, the number of days required for larvae to complete their development in well-grown plants ranged from 11–14 days at 25° to 55–80 days at 5°. This corresponds to an accumulated temperature of approximately 286 day degrees C above the developmental threshold of 0.5°.The daily rate of development of larvae was marginally most rapid between 12° and 18°. When larvae were feeding in small, poorly growing plants, development was delayed and more shoots were needed before pupation than for healthy plants.The pupal stage which develops in the field from the end of April to June needs approximately 400 and 420 day degrees for completion in males and females (threshold 5°). Total day degrees required for the hatching of the egg to adult are about 700 (males) and 720 (females).Records of maximum and minimum temperatures from a meteorological screen 1 m above the ground can be used to calculate the number of day degrees accumulated from 1 May to 12 June, from which the dates of peak emergence of wheat bulb fly can be predicted. If 350 day degrees or more accumulate during this period, the peak emergence occurs near 20 June and with 200 or fewer day degrees it occurs near 11 July.The amount of damage to wheat by wheat bulb fly larvae in 1953, 1954, 1965 and 1966 depended largely on the number of eggs laid, the date of sowing, and also on the rate at which temperature accumulated in the autumn and winter; in all years, late sown crops would have had little opportunity to grow beyond the susceptible stage by the time they were attacked.

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Résumé Quand des jeunes plants de blé d'hiver en pots sont contaminés avec des oeufs de Delia coarctata et maintenus à des températures respectives de 5°, 8°, 10°, 12°, 15° et 20° C, seulement 14% à 44% de ces oeufs donnent des larves qui parviennent á attaquer les plants, alors que si on utilise directement des larves nouveau-nées on obtient 54 à 92% d'attaques réussies. Aux températures de 5° à 25° le temps nécessaire aux larves pour achever leur développement dans les plants bien développés avant de gagner le sol pour la pupaison, est de 11 à 14 jours à 25° contre 55 à 80 jours à 5°.Une courbe de la vitesse de développement établie à partir de la durée du stade larvaire de l'éclosion à la pupaison aux differentes températures montre que le développement est le plus rapide entre 12° et 18° et que le zéro de développement est aux alentours de 0,5°. Le nombre de degrésjours accumulés par la larve de l'éclosion à la pupaison est d'environ 286. Quand les larves se nourrissent aux dépens de plants petits à croissance faible, leur développement est retardé et exige plus de temps et plus de plantes avant la pupaison que lorsqu'elles se nourrissent sur des plants vigoureux.Le stade pupal qui intervient dans la nature de fin avril à juin, nécessite approximativement 400 à 420 degrés-jours pour le développement du mâle et de la femelle au-dessus d'une supposée température de base de 5°. Le nombre total de degrés-jours de l'oeuf à l'adulte est respectivement de 686 et 706 pour le mâle et la femelle.Les enregistrements des températures maxima et minima dans un abri météorologique à 1 m audessus du sol peuvent être utilisés pour établir le nombre de degrés-jours accumulés du 1 er'mai au 12 juin; à partir de ces données les pics d'émergence de D. coarctata peuvent être prédits. Si 350 degrésjours ou plus sont accumulés durant cette période, le pic d'émergence apparaît vers le 20 juin, mais si 200 degres-jours au moins sont notés le pic sera proche du 11 juillet.L'importance des dégâts occasionnés au bleî d'hiver en Angleterre par les larves de D. coarctata en 1953, 1954, 1965 et 1966 dépend largement du nombre d'oeufs pondus, de la date du semis et de la température accumulée en automne et en hiver; au cours de chacune de ces années les cultures à semis tardif auraient eu peu de possibilité de dépasser le stade sensible avant le moment où elles ont été attaquées.