Isolation and fractionation of CHO chromosomes in aqueous two phase systems using charged polymers and base specific macroligands

Summary Chromosomes were isolated in a preparative scale by synchronisation of CHO cells with a double Thymidine block followed by an arrest in the metaphase by addition of Colcemid. Under proper cultivation conditions a mitotic index of 77% total cells could be routinely achieved. Bulk chromosome preparations free of nuclei and other subcellular particles have been obtained by low speed centrifugation followed by a 60 transfer countercurrent distribution using aqueous two phase systems composed of polyethylenglycol and dextran. The partition of CHO chromosomes previously purified in aqueous two phase systems were studied further to develop a protocol for the separation and isolation of individual chromosomes. Partition experiments with chromosomes changing the electrostatic phase potential by addition of charged PEG-derivatives suggest the existence of relatively highly charged chromosome groups. Most promising results with regard to separation were obtained using two PEG-derivatives, which interact specifically with the bases in DNA. For this affinity partitioning a GC- and AT-specific macroligand were employed. Comparing CCD's using each of these ligands information on the GC and AT content of exposed DNA in the chromosomes groups could be derived, demonstrating that specific sequences of DNA are accessible at the surface of metaphase chromosomes.     

Isolation of a brain peptide identical to the intestinal PHI (peptide HI)

The isolation of a brain peptide identical to the intestinal peptide PHI (peptide HI) is described. The peptide was isolated from porcine brain extract using a chemical assay method based on its C-terminal isoleucine amide structure. The complete amino acid sequence of the peptide was found to be: His-Ala-Asp-Gly-Val-Phe-Thr-Ser-Asp-Phe-Ser-Arg-Leu-Leu-Gly-Gln-Leu-Ser-Ala- Lys-Lys-Tyr-Leu-Glu-Ser-Leu-Ile-NH2. This sequence is identical to the intestinal peptide thus demonstrating PHI to be a brain-gut peptide. The role of PHI in the central nervous system as a neurotransmitter or neuromodulator is discussed.     

渗透胁迫对水稻幼苗膜脂过氧化及体内保护系统的影响

两个不同抗旱性的水稻品种对PEG6000渗透胁迫(-0.5MPa,-0.8MPa)的反应具有一定差异。渗透胁迫下SOD,POD和CAT活性及Vc,Car含量与膜脂过氧化水平及膜透性呈一定负相关性,表明这些指标可作为水稻抗旱育种的参考依据。     

亚洲产Clavicorona的形态学及亲和性研究

中国产的Clavicorona pyxidata与北美的种样本间配对实验结果显示该种在不同的地理分布区内的种群之间完全性亲和或具有相同的交配型等位基因,表明中国产与北美产的该形态学种同属于一个生物种。日本产的标本与C. pyxidata在子实体外部形态及孢子尺度上有差异,虽然在日本被鉴定为该种。日本产的菌株与C. pyxidata菌株间配对实验证明二者之间完全不亲和,即属于不同的生物种。生殖隔离拌随着形态学变异。     

Simulated and measured nitrogen conditions in a manured and fertilised soil

The governing factors for soil nitrogen dynamics were identified with a simulation model. In addition, the model was used to interpret measurements from a plot fertilisation experiment in southwest Sweden.Simulated moisture and temperature conditions were the driving variables for the simulation of soil nitrogen dynamics and leaching during a 6-year period. The results of the simulation were compared with monthly observations on two plots with grain crops, one with liquid manure and commercial fertilisers applied and one with commercial fertilisers only.Simulated temporal variations of the nitrate and ammonium storages generally agreed with observations. The dominant role of the crops as a determinant of soil nitrogen conditions was demonstrated. A higher leaching loss from the plot with application of commerical fertilisers only occurred both in simulations and measurements compared to the plot with application of both commercial fertilisers and manure. The main reason was the higher N-application in the former treatment.The effect of water flows in macropores was interpreted as a delay of simulated leaching compared to observed leaching on some occasions in summer and early autumn. No direct effect of the macropores on the yearly rates of leaching could be seen.     

Numbers of sporophytic self-incompatibility alleles in populations of wild radish

Summary To estimate the numbers of sporophytic S-alleles in two adjacent populations of wild radish, we performed 701 reciprocal crosses among 50 individuals. Each cross was replicated five times in each direction. Sixteen plants were fully intercompatible, indicating the presence of at least 32 S-alleles in the two populations. A minimum of 22 S-alleles occur in a single population. The frequency of incompatibility was significantly higher for within-population crosses (14.5%) than for between-population crosses (7.8%). This suggests that the two populations differ in the composition and frequency of alleles at the S-locus.     

A new microcomputer-controlled neutron activation and analysis system

A microcomputer-controlled irradiation and measurement system and a microprocessor-controlled sample changer have been installed at the SLOWPOKE-2 Facility at the Royal Military College of Canada (RMC). These systems can provide the gamut of instrumental neutron activation analysis (INAA) techniques for the analyst. Custom software has been created for system control, data acquisition, and off-line spectral analysis using programs that incorporate Gaussian peak-fitting methods of analysis. The design and use of the equipment is discussed, and the performance is illustrated with results obtained from the analysis of marine sediment and biological reference materials.     

Analysis of time-resolved fluorescence anisotropy in lipid-protein systems

Fluorescent probes located in heterogeneous environments give rise to anomalous time-resolved fluorescence anisotropy. A simple analytical expression of anisotropy has been derived for the case of a small difference in local fluorescence lifetimes. The expression has the diagnostic advantage that the time dependence of the fluorescence anisotropy can be predicted from the differences in fluorescence lifetimes and residual anisotropies of the probes located in different sites. Using this model, the local fluorescence anisotropy parameters and the relative contributions of the lipid probe octadecyl rhodamine B in a lipid environment and in the vicinity of bacteriophage M13 coat protein reconstituted in phospholipid bilayers, composed of 80% 1,2-dimyristoyl-sn-glycero-3-phosphocholine and 20% 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol have been determined experimentally. At 40°C, the correlation times for bound and free probes are 2.3 and 3.0 ns, respectively, while the corresponding order parameters are 0.85 and 0.62, respectively.Abbreviations ESR electron spin resonance - DMPC 1,2-dimyristoyl-sn-glycero-3-phosphocholine - DMPC 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol - L/P ratio phospholipid to coat protein molar ratio - <> average fluorescence lifetime - r(0) initial anisotropy - r() residual anisotropy On leave of Shanghai Medical Equipment Research Institute, 77 Jiang Ning Rd. Shanghai, People's Republic of China Offprint requests to: M. A. Hemminga     

Growth and development of biofeedback: A bibliographic update

Computerized literature searching techniques were used to examine publication patterns in the worldwide biofeedback literature. Searches were completed in the United States and in Japan for the years 1985 through 1987. The results were used to update the results of an earlier study (Hatch & Riley, 1985) that covered the years from 1964 through 1984. Publication growth curves were generated for several media, including scientific journal articles, books, doctoral dissertations, and popular magazine articles. Although publication of information about biofeedback remains active worldwide, there has been a declining trend in effect for the past several years. The American database grossly underestimated the number of Japanese biofeedback articles, and it is likely that the literatures of other countries outside of North America were similarly underestimated. Possible explanations for the various trends noted are discussed.These results were presented in part at the Twentieth Annual Meeting of the Association for Applied Psychophysiology and Biofeedback. We gratefully acknowledge the assistance of Margaret Cyr-Provost in preparing the data for analysis.     

Comparative character of the deep-water and inshore cottoid fishes endemic to Lake Baikal

Lake Baikal's 29 endemic species of cottoid fishes form three groups: shallow-water species in depths to 350 m; eurybathic species from 50 to 1300 m; and abyssal species from 400 to 1600 m. These groups differ in their abilities to withstand high hydrostatic pressure. As in marine deep-water fishes, abyssal cottoids in Baikal have few or no cones in the retina, and some have tubular eyes. Their seismosensory systems predominate, based chiefly on free neuromasts. The proportion of species with canal systems decreases with depth. Diversity of the predominantly gammarid foods also decreases from 45 species in shallow water to five species in deep water, and the lateral line system plays the dominant role in food detection at all depths. Two abyssal cottoids have become secondary pelagic, achieving close to neutral buoyancy through high lipid levels and reduced skeletal mineralization. These forms take advantage of the abundant pelagic planktonic amphipod populations. The adaptations of abyssal forms parallel those seen in deep-water marine fishes.