多菌灵在黄精根茎无菌培养中的应用

本研究中比较了几种杀菌剂对黄精根茎的灭菌效果。结果表明,采用乙醇、饱和漂白粉、HgCI2等杀菌剂对黄精根茎杀菌时,微生物的污染率很高;先用3%多菌灵溶液对黄精根茎进行内吸杀菌处理45h以上,然后再用0·2%HgCI2进行灭菌10min,则黄精根茎的污染率可得到有效控制。多菌灵的处理效果与黄精根茎的大小、类型和年龄等因素有关。     

具污染与捕获的Logistic单种群的持续生存及绝灭

当今社会工农业的高速发展在给人们带来经济利益的同时,也造成了严重的环境污染．一些生物种群一方面受到污染的威胁,另一方面还要满足人们捕获的需求．这就使得环境污染中的种群捕获问题成为人们关心的问题．本文指出以往环境污染单种群模型的不足之处,基于文献[3]的基础上给出新模型;并得到了一致持续生存及灭绝的充分条件;而且通过具体例子给出两个控制变量：环境毒素输入率和捕获率之间的关系图．     

Respective importance of protein folding and glycosylation in the thermal stability of recombinant feruloyl esterase A

The thermal stability of four molecular forms (native, refolded, glycosylated, non-glycosylated) of feruloyl esterase A (FAEA) was studied. From the most to the least thermo-resistant, the four molecular species ranked as follows: (i) glycosylated form produced native, (ii) non-glycosylated form produced native, (iii) non-glycosylated form produced as inclusion bodies and refolded, and (iv) glycosylated form produced native chemically denatured and then refolded. On the basis of these results and of crystal structure data, we discuss the respective importance of protein folding and glycosylation in the thermal stability of recombinant FAEA.     

Temperature-related mortality in France, a comparison between regions with different climates from the perspective of global warming

This paper aims to explain the results of an observational population study that was carried out between 1991 and 1995 in six regions (departments) in France. The study was to assess the relationship between temperature and mortality in a few areas of France that offer widely varying climatic conditions and lifestyles, to determine their thermal optimum, defined as a 3 degrees C temperature band with the lowest mortality rate in each area, and then to compare the mortality rates from this baseline band with temperatures above and below the baseline. The study period was selected because it did not include extreme cold or hot events such as a heatwave. Data on daily deaths from each department were first used to examine the entire population and then to examine men, women, various age groups and various causes of death (respiratory disease, stroke, ischaemic heart disease, other disease of the circulatory system, and all other causes excluding violent deaths). Mean temperatures were provided by the National Weather Service. The results depicted an asymmetrical V- or U-shaped relationship between mortality and temperature, with a thermal optimum lower for the elderly, and generally lower for women than for men except in Paris. The relationship was also different depending on the cause of death. In all cases, more evidence was collected showing that cold weather was more deadly than hot weather, and it would now be interesting to enlarge the study to include years with cold spells and heatwaves. Furthermore, the results obtained could be of great use in estimating weather-related mortality as a consequence of future climate-change scenarios.     

A comparison of growing season indices for the Greater Baltic Area

Predictions of the effects of global warming suggest that climate change may have large impacts on ecosystems. The length of the growing season is predicted to increase in response to increasing global temperatures. The object of this study was to evaluate different indices used for calculating the thermal growing season for the Greater Baltic Area (GBA). We included established indices of growing season start, end and length, as well as new and modified indices. Based on the results, the GBA can be divided into a maritime western part and a more continental eastern part, with the western part reacting more sensitively to the use of different indices. The eastern part is more stable, but even here the index-to-index differences are large. It was found that including or excluding a frost criterion had a significant influence on the initiation of the growing season in the western, maritime, parts of the GBA. Frost has not the same importance for the end of the growing season. However, some end indices can result in a “never ending” growing season. When looking at twentieth century trends in growing season parameters, it was found that, when averaged over the whole GBA, there was little difference in trends depending on the indices used. The general mean trend in the GBA for the twentieth century discloses an earlier onset of c. 12 days, a delayed end of c. 8 days and consequently a lengthening of the growing season of about 20 days.     

Structural insights into the effect of isonucleosides on B-DNA duplexes using molecular-dynamics simulations

Some structural insights into the conformations of the isonucleosides containing duplexes have been provided. Unrestrained molecular-dynamics simulations on 18-mer duplexes with isonucleosides incorporated at the 3'-end or in the center of one strand have been carried out with explicit solvent under periodic boundary conditions using the AMBER force field and the particle mesh Ewald method. The Watson–Crick hydrogen-bonding patterns of the duplexes studied remained intact throughout the simulation. For the modified duplexes, the changes observed in the inter-base pair parameters and backbone torsional angles were primarily localized at the isonucleoside-inserted area. All five structures studied remained in the B-form family. The decreased stacking abilities indicated by the large changes in inter-base pair parameters and the large changes in backbones made the modified duplexes show a minor thermal destabilization in comparison with native DNA. The MM_PBSA method for estimating binding free energies on two complementary strands was used. The results showed that the binding free energies of isonucleoside-incorporated DNA duplexes were lower than the native DNA duplex, which is in good agreement with experimental observations.      

Structural basis of the destabilization produced by an amino-terminal tag in the beta-glycosidase from the hyperthermophilic archeon Sulfolobus solfataricus

We have previously shown that the major ion-pairs network of the tetrameric beta-glycosidase from the hyperthermophilic archeon Sulfolobus solfataricus involves more than 16 ion-pairs and hydrogen bonds between several residues from the four subunits and protects the protein from thermal unfolding by sewing the carboxy-termini of the enzyme. We show here that the amino-terminal of the enzyme also plays a relevant role in the thermostabilization of the protein. In fact, the addition of four extra amino acids at the amino-terminal of the beta-glycosidase, though not affecting the catalytic machinery of the enzyme and its thermophilicity, produced a faster enzyme inactivation in the temperature range 85-95 degrees C and decreased the Tm of the protein of 6 degrees C, measured by infrared spectroscopy. In addition, detailed two-dimensional IR correlation analysis revealed that the quaternary structure of the tagged enzyme is destabilized at 85 degrees C whilst that of the wild type enzyme is stable up to 98 degrees C. Molecular models allowed the rationalization of the experimental data indicating that the longer amino-terminal tail may destabilize the beta-glycosidase by enhancing the molecular fraying of the polypeptide and loosening the dimeric interfaces. The data support the hypothesis that fraying of the polypeptide chain termini is a relevant event in protein unfolding.     

Length and pH-dependent energetics of (CCG)n and (CGG)n trinucleotide repeats

Trinucleotide repeats are involved in a number of debilitating diseases such as fragile-X syndrome and myotonic dystrophy. Eighteen to 75 base-long (CCG)(n) and (CGG)(n) oligodeoxynucleotides were analysed using a combination of biophysical (UV-absorbance, differential scanning calorimetry) and biochemical methods (non-denaturing gel electrophoresis, enzymatic footprinting). All oligomers formed stable intramolecular structures under near physiological conditions with a melting temperature which was only weakly dependent on oligomer length. Thermodynamic analysis of the denaturation process by UV-melting and calorimetric experiments revealed a length-dependent discrepancy between the enthalpy values deduced from model-dependent (UV-melting) and model-independent experiments (calorimetry), as recently shown for CTG and CAG trinucleotides (Nucleic Acids Res. 33 (2005) 4065). Evidence for non-zero molar heat capacity changes was also derived from the analysis of the Arrhenius plots. Such behaviour is analysed in the framework of an intramolecular "branched" or "broken" hairpin model, in which long oligomers do not fold into a simple long hairpin-stem intramolecular structure, but allow the formation of several independent folding units of unequal stability. These results suggest that this observation may be extended to various trinucleotide repeats-containing sequences.     

A recombinant transductor-effector system: in vitro study of G inhibitory protein (G-alpha-i1) direct activators

Most protease prosegments are co-synthesized at the N-termini of cysteine proteases and are involved in folding assistance, inhibition, and activation of their mature enzymes. By using circular dichroism, UV-difference and fluorescence spectroscopies, we studied the thermal unfolding of papain prosegment. The transition seems to be two-state and reversible, with an unfolded state prone to aggregation. Unfolding thermodynamic parameters obtained show low values both for deltaH(Tm) and deltaCp(U), indicative of a loosely packed three-dimensional conformation for the prosegment at near-neutral pH conditions. In spite of these results, fluorescence experiments demonstrate that papain prosegment is able to recognize and inhibit its cognate protease. An acid medium induces a molten globule-like state without intermediates, which in turn undergoes an irreversible thermal unfolding. Our results suggest that papain prosegment has a high degree of conformational flexibility, with the ability to form not only a molten globule-like structure in activating conditions, but also requiring an induced fit in order to be functional as inhibitor.