Effects of proteins on the thermotropic phase transitions of phospholipid membranes

A variety of proteins have been studied for their ability to interact and alter the thermotropic properties of phospholipid bilayer membranes as detected by differential scanning calorimeter. The proteins studied included: basic myelin protein (A1 protein), cytochrome c, major apoprotein of myelin proteolipid (N-2 apoprotein), gramicidin A, polylysine, ribonuclease and hemoglobin. The lipids used for the interactions were dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylglycerol. The interactions were grouped in three categories each having very different effects on the phospholipid phase transition from solid to liquid crystalline. The calorimetric studies were also correlated with data from vesicle permeability and monolayer expansion.Ribonuclease and polylysine which exemplify group 1 interactions, show strong dependence on electrostatic binding. Their effects on lipid bilayers include an increase in the enthalpy of transition (ΔH) accompanied by either an increase or no change in the temperature of transition (T_c). In addition, they show minimal effects on vesicle permeability and monolayer expansion. It was concluded that these interactions represent simple surface binding of the protein on the lipid bilayer without penetration into the hydrocarbon region.Cytochrome c and Al protein, which exemplify group 2 interactions, also show a strong dependence on the presence of net negative charges on the lipid bilayers for their binding. In contrast to the first group, however, they induce a drastic decrease in both T_c and ΔH of the lipid phase transition. Furthermore, they induce a large increase in the permeability of vesicles and a substantial expansion in area of closely packed monolayers at the air-water interface. It was concluded that group 2 interactions represent surface binding followed by partial penetration and/or deformation of the bilayer.Group 3 interactions, shown by proteolipid apoprotein and gramicidin A, were primarily non-polar in character, not requiring electrostatic charges and not inhibited by salt and pH changes. They had no appreciable effect on the T_c but did induce a linear decrease in the magnitude of the ΔH, proportional to the percentage of protein by weight. Membranes containing 50% proteolipid protein still exhibited a thermotropic transition with a ΔH one half that of the pure lipid, and only a small diminution of the size of the cooperative unit. It was concluded that in this case the protein was embedded within the bilayer, associating with a limited number of molecules via non-polar interactions, while the rest of the bilayer was largely unperturbed.     

Simultaneous tissue profiling of eicosanoid and endocannabinoid lipid families in a rat model of osteoarthritis

We describe a novel LC method for the simultaneous and quantitative profiling of 43 oxylipins including eicosanoids, endocannabinoids, and structurally related bioactive lipids with modified acyl groups. The LC-MS/MS method uses switching at a defined time between negative and positive electrospray ionization modes to achieve optimal detection sensitivity for all the lipids. The validated method is linear over a range of 0.01–5 nmol/g (0.1–50 nmol/g for 2-arachidonoyl glycerol) with intra- and interday precision and accuracy between 1.38 and 26.76% and 85.22 and 114.3%, respectively. The method successfully quantified bioactive lipids in different tissue types in the rat, including spinal cord, dorsal root ganglia (DRGs), knee joint, brain, and plasma. Distinct regional differences in the pattern of lipid measured between tissue types were observed using principle component analysis. The method was applied to analyze tissue samples from an established preclinical rat model of osteoarthritis (OA) pain and showed that levels of 12-hydroxyeicosatetraenoic acid were significantly increased in the OA rat knee joint compared with controls, and that 15-hydroxyeicosatetraenoic acid was significantly increased in the DRGs in the model of OA compared with controls. The developed LC-MS/MS method has the potential to provide detailed pathway profiling in tissues and biofluids where the disruption of bioactive oxylipins may be involved in disease states.     

Short-term effects of temperature enhancement on growth and reproduction of alpine grassland species

In order to study the effects of temperature enhancement on alpine calcareous grassland species, a warming experiment was carried out in the Berchtesgaden National Park (Southeast Germany, Northern Calcareous Alps) between 2002 and 2004. The study was conducted in stands of the Carex sempervirens and the Carex firma communities; the two most widespread grassland types in the alpine zone of the Northern Calcareous Alps. The temperature of the vegetation stand and the upper soil was passively enhanced using open top chambers (OTCs). The construction of the OTCs was appropriate since temperature was clearly increased while water conditions (humidity, soil water content) were not changed.

By comparing manipulated (temperature enhancement) with non-manipulated plots, the effects of warming on growth and reproduction of selected key species were studied. To test if vegetation response to temperature enhancement is at least partly due to increases in nutrient availability, soil solution concentrations of nitrate and ammonium were analysed.

We found that most of the studied plant species are sensitive to temperature enhancement. Growth and/or reproduction of 12 of the 14 studied species were significantly stimulated by warming. Only two species showed no response; none of the species experienced decreases in growth or reproduction. Dwarf shrubs and graminoids showed a stronger response than herbaceous perennials. A significant effect of warming on nutrient availability could not be detected. The observed response of vegetation is therefore mainly caused by direct and not by indirect temperature effects.     

Nocturnal parasitism of moth eggs by Trichogramma wasps

Parasitoid wasps of the genus Trichogramma are used worldwide as biological control agents against lepidopteran pests. Trichogramma wasps develop inside eggs of a wide range of host species, most of them moths. They are generally considered as diurnal insects. Here, we investigated whether Trichogramma wasps can also successfully parasitise host eggs at night under controlled laboratory conditions. Eggs of the moth Ephestia kuehniella were offered under dark conditions (scotophase) to females of Trichogramma brassicae and Trichogramma evanescens either from 9:00 PM to 9:00 AM or from 11:00 AM to 5:00 PM at four different temperatures (5°C, 10°C, 15°C and 20°C). Both species are known to parasitise E. kuehniella eggs in the photophase during daytime. The results show that T. brassicae did not parasitise eggs in the scotophase at night and only very few in the artificially induced scotophase during daytime from 10°C to 20°C. In contrast, T. evanescens parasitised more eggs in the dark both at night and artificially induced scotophase during daytime. Parasitism in the scotophase already started at 5°C, with more eggs being parasitised and more offspring being produced at higher temperatures. T. evanescens displayed higher parasitism activity in the induced scotophase during daytime than in the scotophase at night. The present study suggests that Trichogramma are capable of successfully parasitising host eggs at night, even at low temperatures, but that nocturnal activity with respect to parasitism varies between wasp species.     

Trap‐shyness subsidence is a threshold function of mark–recapture interval in brown mudfish Neochanna apoda populations

The influence of capture interval on trap shyness, and temperature, rainfall and drought on capture probability (p) in 827 brown mudfish Neochanna apoda was quantified using mark–recapture models. In particular, it was hypothesized that the loss of trapping memory in marked N. apoda would lead to a capture‐interval threshold required to minimize trap shyness. Neochanna apoda trap shyness approximated a threshold response to capture interval, declining rapidly with increasing capture intervals up to 16·5 days, after which p remained constant. Tests for detecting trap‐dependent capture probability in Cormack–Jolly–Seber models failed to detect trap shyness in N. apoda capture histories with capture intervals averaging 16 days. This confirmed the applicability of the 16 day capture‐interval threshold for mark–recapture studies. Instead, N. apoda p was positively influenced by water temperature and rainfall during capture. These results imply that a threshold capture interval is required to minimize the trade‐off between the competing assumptions of population closure and p homogeneity between capture occasions in closed mark–recapture models. Moreover, environmental factors that influence behaviour could potentially confound abundance indices, and consequently abundance trends should be interpreted with caution in the face of long‐term climate change, such as with global warming.     

High-level expression of murine terminal deoxynucleotidyl transferase inEscherichia coli grown at low temperature and overexpressingargU tRNA

Terminal deoxynucleotidyl transferase (TdT) is a highly conserved vertebrate enzyme that possesses the unique ability to catalyze the random addition of deoxynucleoside 5′-triphosphates onto the 3′-hydroxyl group of a single-stranded DNA. It plays an important role in the generation of immunoglobin and T-cell receptor diversity. TdT is usually obtained from animal thymus gland or produced in a baculovirus system, but both procedures are rather tedious, and proteolysis occurs during purification. Attempts to overexpress TdT in bacteria have been unsuccessful or have yielded an enzyme with a lower specific activity. A dearth of TdT has thus hampered detailed structural and functional studies. In the present study, we report that by lowering growth temperature and overexpressing a rare arginyl tRNA, it is possible to boost the production inEscherichia coli of murine TdT with minimal proteolysis and high specific activity.     

The Influences of Environment, Mating Habitat, and Predation on Evolution of Pinniped Lactation Strategies

Seals have adapted their social systems and lactation strategies to marine environments that include open and ice-covered oceans, high and low productivity, extremes in seasonality, and ocean- and terrestrial-type predators. Different explanations for the variation in pinniped lactation systems have been proposed but tests of alternative hypotheses have not sufficiently accounted for phylogeny and body size. After controlling for this variation, I predicted that environment, mating habitat, and predation would yield a fuller explanation. Lactation traits, duration, pup growth rate, and fat content were significantly influenced by both body size and phylogeny, which together explained 20–69% of the variation. After controlling for this variation, initial results did not support the environment hypothesis, as no differences in lactation traits were found between species living in polar (≥60°N) versus equatorial (<60°N) environments. In contrast, seals that nurse in areas of Arctic sea ice contending with ice-hunting predators, such as polar bears, had relatively short lactation compared to species living in the Antarctic and more equatorial regions. Also, the availability of predator-free islands for terrestrial mating and parturition was related to a harem mating system, increased sexual size dimorphism (SSD), and slow juvenile growth rates, less fat in milk, and longer lactation. Using structural equation modeling, latitude and size of harems provided independent explanations for all three lactation traits. Thus, use of islands in ice-free waters, predation in Arctic ice-covered waters, and more milk fat in high-latitude seals together provided adequate explanations for the evolution of lactation diversity among pinnipeds.     

Electric discharge plasmas influence attachment of cultured CHO K1 cells

Non-thermal plasmas can be generated by electric discharges in gases. These plasmas are reactive media, capable of superficial treatment of various materials. A novel non-thermal atmospheric plasma source (plasma needle) has been developed and tested. Plasma appears at the end of a metal pin as a submillimetre glow. We investigate the possibility of applying the plasma needle directly to living tissues; the final goal is controlled cell treatment in microsurgery. To resolve plasma effects on cells, we study cultured Chinese hamster ovarian cells (CHO-K1) as a model system. When these are exposed to the plasma, instantaneous detachment of cells from the surface and loss of cell-cell interaction is observed. This occurs in the power range 0.1-0.2 W. Cell viability is assessed using propidium iodide (PI) and cell tracker green (CTG) fluorescent staining utilizing confocal laser scanning microscopy (CLSM). Detached cells remain alive. Use of higher doses (plasma power >0.2 W) results in cell necrosis. In all cases, plasma-influenced cells are strictly localized in submillimetre areas, while no reaction in surrounding cells is observed. Due to its extreme precision, plasma treatment may be applicable in refined tissue modification.     

不同抗凝人血浆凝固酶检测的探讨

目的 了解3种不同抗凝人血浆检测血浆凝固酶的可行性及观察时间对结果的影响.方法 用3种不同抗凝人血浆及以不同菌液浓度作试管法凝固酶试验.结果 EDTA-K2抗凝血浆出现阳性慢,凝块大多较小,24 h内稳定性较好;枸橼酸钠抗凝血浆出现结果介于EDTA-K2、肝素抗凝血浆之间,结果最不稳定;肝素抗凝血浆出现阳性最快,但不够稳定;菌液浓度对出现阳性的早晚无影响.结论 3种抗凝人血浆在试管法凝固酶测定中使用时,出现阳性快慢不受菌液浓度影响,随时间延长溶解现象可能会增加,应掌握结果观察时间,以降低假阴性.