Solution structure by 1H and dynamics by natural abundance 13C NMR of a receptor recognising peptide derived from a C-terminal fragment of neuropeptide Y

Summary A peptide consisting of 20 amino acid residues, derived from a C-terminal fragment of neuropeptide Y (NPY) and showing high affinity to NPY receptors, was synthesised. Its sequence is PAADLARYRHYIN-LITRQRY-NH₂, and the solution structure was calculated from NMR-derived distance and torsion angle restraints, obtained at 15°C in a solvent mixture of water and 30% (v/v) 1,1,1,3,3,3-hexafluoro-2-propanol, by using DIANA and restrained energy minimisation. The structure was found to consist of a well-defined -helix in the centre, with a few residues at the termini having less well defined conformations. The spinlattice and spin-spin relaxation rates of -carbons have been determined on ¹³C at natural abundance. From 1D experiments the global rotational correlation time was determined and from 2D experiments the dynamics of each individual residue was obtained. The results demonstrate that the C-H vectors in the -helix essentially follow the global motion. Towards the termini, contributions from local dynamics increase. This tendency is correlated to the increasing uncertainty of the structure towards the peptide ends. An effective molecular volume was calculated from the temperature dependence of the global rotational correlation time. This is well compatible with a monomeric peptide, solvated by water and 1,1,1,3,3,3-hexafluoro-2-propanol. The presence of peptide dimers was ruled out as being inconsistent with the relaxation data.Supplementary material available from the authors: Two data tables and 10 PDB coordinate files of the calculated NMR structures of P7. One data table contains all detected and integrated NOE intensities; the other connects each proton and pseudoatom to an atom number used in the NOE table. The table contents served as input data files for CALIBA.Currently on leave from the Institute of Chemical Physics and Biophysics, Tallinn, Estonia.     

Detention and mixing in free water wetlands

Mixing was studied in free water surface wetland receiving pumped river water, by measurement of the non-interacting tracer lithium. The flow pattern was found to be intermediate between plug flow and well-mixed. The nominal detention time, calculated from volume aand flow, was 50% larger than the mean tracer detention time. The peak time was found to be one-half the tracer detention time. Three models were constructed: plug flow with dispersion, tanks in series, and a series-parallel network of tanks. All proved capable of fitting the exit tracer concentration curves but the network model provided a better fit to internal measurements. Pumping frequency was high enough to allow use of an average flowrate. The degree of mixing, as characterized by the variance of the exit tracer response curve, was comparable to that found by other researchers for wetlands, ponds and rivers.     

解离增强镧系元素荧光免疫分析灵敏度的改进

为提高解离增强镧系荧光免疫分析(DELFIA)的灵敏度或信／噪比,进行了一些重要的方法学研究．观察到了对于不同的铕量,荧光响应和信／噪比都随着增强液体积而明显地变化．对于确定的铕量,存在一个最佳体积,且铕量越小,其最佳体积也越小．实验中选择最佳体积是重要的．研究了增强液制备技术,发展了微滴定板条有效的清洗和干燥方法,使本底荧光明显降低．     

The effect of pulsed microwaves on passive electrical properties and interspike intervals of snail neurons

The effects of pulsed microwaves (2.45 GHz, 10 μs, 100 pps, SAR: 81.5 kW/kg peak, 81.5 W/kg average) on membrane input resistance and action potential (AP) interval statistics were studied in spontaneously active ganglion neurons of land snails (Helix aspersa), at strictly constant temperature (20.8±.07°C worst case). Statistical comparison with sham-irradiated neurons revealed a significant increase in the mean input resistance of neurons exposed to pulsed microwaves (P ? .05 ). Pulsed microwaves had no visible effect on mean AP firing rate; this observation was confirmed by analysis of interspike intervals (ISIs). Using an integrator model for spontaneously active neurons, we found the net input current to be more variable in neurons exposed to pulsed microwaves. The mean input current was not affected. The standard deviation of ISIs and the autocorrelation of the input current were marginally affected, but these changes were not consistent across neurons. Although the observed effects were less obvious than those reported in other studies, they represent evidence of a direct interaction between neurons and pulsed microwaves, in the absence of macroscopic temperature changes. The data do not suggest a single, specific mechanism for such interaction. © 1993 Wiley-Liss, Inc.     

Effective population size,genetic diversity,and coalescence time in subdivided populations

A formula for the effective population size for the finite island model of subdivided populations is derived. The formula indicates that the effective size can be substantially greater than the actual number of individuals in the entire population when the migration rate among subpopulations is small. It is shown that the mean nucleotide diversity, coalescence time, and heterozygosity for genes sampled from the entire population can be predicted fairly well from the theory for randomly mating populations if the effective population size for the finite island model is used.     

Overexpression of cloned genes using recombinant Escherichia coli regulated by a T7 promoter: II. Two-stage continuous cultures and model simulations

A two-stage culture strategy was studied for continuous high-level production of a foreign protein in the chemically inducible T7 expression system. The first stage is dedicated to the maintenance of plasmid-bearing cells and the second stage to the target protein synthesis by induction of cells coming from the first stage. On entering the second stage, recombinant cells undergo a gradual induction of the target gene expression. These plasmid-bearing cells experience dynamic changes in intracellular compositions and specific growth rates with their individual residence times. Therefore, the overall cultural characteristics in the production stage are really averages of the contributions from the various cells with different residence times. The behavior of the two-stage culture is described by a model, which accounts for dynamic variations of cell growth and protein synthesis rates with cell residence times. Model simulations were compared with experimental results at a variety of operating conditions such as inducer concentration and dilution rate. This model is useful for understanding the behavior of two-stage continuous cultures. (c) 1993 John Wiley & Sons, Inc.     

Variable generation times and Darwinian fitness measures

Summary Reproductive value (RV) and net reproductive output (R _o) are frequently used fitness measures. We argue that they are only appropriate when intervals between reproductive events are fixed, as they are dimensionless generation-to-generation scalings with units offspring per parent. A fitness measure should account for two different effects of a decrease in generation time: (1) increased survival due to shorter exposure to mortality agents and (2) increased frequency of reproduction.R _o andRV deal with the first of these two effects, while a measure with a physical dimensionper time [T^–1] is needed to account for the second. The Malthusian growth parameter,r, meets this requirement and in situations where time to reproduction is variable, we propose, the instantaneous rate of spread of descendants (from an individual) be used instead ofR _o. As an alternative toRV, we suggest using the instantaneous difference = –r, wherer is the population rate of increase. WhileRV andR _o are dimensionless ratios, , and areper time rates which are appropriate in accounting for alterations in generation time.     

Heat tolerance of marine ectotherms in a warming Antarctica

Global warming is affecting the Antarctic continent in complex ways. Because Antarctic organisms are specialized to living in the cold, they are vulnerable to increasing temperatures, although quantitative analyses of this issue are currently lacking. Here we compiled a total of 184 estimates of heat tolerance belonging to 39 marine species and quantified how survival is affected concomitantly by the intensity and duration of thermal stress. Species exhibit thermal limits displaced toward colder temperatures, with contrasting strategies between arthropods and fish that exhibit low tolerance to acute heat challenges, and brachiopods, echinoderms, and molluscs that tend to be more sensitive to chronic exposure. These differences might be associated with mobility. A dynamic mortality model suggests that Antarctic organisms already encounter temperatures that might be physiologically stressful and indicate that these ecological communities are indeed vulnerable to ongoing rising temperatures.     

G418对IL—2基因包装细胞的筛选

本文用高浓度的G418(800μg/ml)和低浓度的G418(200μg/ml)对包装细胞PLXSN/IL-2/PA317细胞进行40天的选择筛选培养,使其细胞呈稳定状态生长时,收集上清液转染NIH/3T3细胞,进行病毒滴度测定。试图在高选择标记的情况下筛选出高表达目的基因的包装细胞。实验结果表明:高、低浓度的G418对PLXSN/IL-2/PA317包装细胞的选择作用相同,即包装细胞的病毒滴度同选择标记物浓度无关。提示可用低浓度的G418来维持包装细胞的生命。     

Ranking species based on sensitivity to perturbations under non-equilibrium community dynamics

Managing ecological communities requires fast detection of species that are sensitive to perturbations. Yet, the focus on recovery to equilibrium has prevented us from assessing species responses to perturbations when abundances fluctuate over time. Here, we introduce two data-driven approaches (expected sensitivity and eigenvector rankings) based on the time-varying Jacobian matrix to rank species over time according to their sensitivity to perturbations on abundances. Using several population dynamics models, we demonstrate that we can infer these rankings from time-series data to predict the order of species sensitivities. We find that the most sensitive species are not always the ones with the most rapidly changing or lowest abundance, which are typical criteria used to monitor populations. Finally, using two empirical time series, we show that sensitive species tend to be harder to forecast. Our results suggest that incorporating information on species interactions can improve how we manage communities out of equilibrium.