Advanced negative detection method comparable to silver stain for SDS-PAGE separated proteins detection

In order to achieve an easy, rapid and sensitive protocol to detect proteins in polyacrylamide gel, an advanced negative detection method comparable to silver stain is described. When a gel was incubated with Phloxine B and followed by the development in acidic solution, the zones where forming protein-dye complex were selectively transparent, unlike opaque gel background. Within 50 min after electrophoresis, down to 0.1–0.4 ng of gel-separated proteins (similar with silver stain) could be observed, without labor-intensive and time-consuming procedure. Comparing with the most common negative stain method, Imidazole-zinc stain, Phloxine B stain has been shown higher sensitivity and distinct contrast between the transparent protein bands/spots and opaque background than those; furthermore, it is no longer necessary to concern about retention time of observation. This technique may provide a sensitive and practical choice for proteomics researches.     

Angiotensin AT2 receptor agonist stimulates high stretch induced- ANP secretion via PI3K/NO/sGC/PKG/pathway

Angiotensin II (Ang II) type 1 receptor (AT₁R) mediates the major cardiovascular effects of Ang II. However, the effects mediated via AT₂R are still controversial. The aim of the present study is to define the effect of AT₂R agonist CGP42112A (CGP) on high stretch-induced ANP secretion and its mechanism using in vitro and in vivo experiments. CGP (0.01, 0.1 and 1 μM) stimulated high stretch-induced ANP secretion and concentration from isolated perfused rat atria. However, atrial contractility and the translocation of extracellular fluid did not change. The augmented effect of CGP (0.1 μM) on high stretch-induced ANP secretion was attenuated by the pretreatment with AT₂R antagonist or inhibitor for phosphoinositol 3-kinase (PI3K), nitric oxide (NO), soluble guanylyl cyclase (sGC), or protein kinase G (PKG). However, antagonist for AT₁R or Mas receptor did not influence CGP-induced ANP secretion. In vivo study, acute infusion of CGP for 10 min increased plasma ANP level without blood pressure change. In renal hypertensive rat atria, AT₂R mRNA and protein levels were up-regulated and the response of plasma ANP level to CGP infusion in renal hypertensive rats augmented. The pretreatment with AT₂R antagonist for 10 min followed by CGP infusion attenuated an increased plasma ANP level induced by CGP. However, pretreatment with AT₁R or Mas receptor antagonist unaffected CGP-induced increase in plasma ANP level. Therefore, we suggest that AT₂R agonist CGP stimulates high stretch-induced ANP secretion through PI3K/NO/sGC/PKG pathway and these effects are augmented in renal hypertensive rats.     

赤霉素对水平放置水曲柳幼苗的负向重性和木材形成的影响

对水平放置2年生水曲柳(Franxinus mandshurica)幼苗应用赤霉素(GAs)及其合成抑制剂烯効唑(Uni．-P)对幼苗的负向重性及木材形成中的生理作用进行了探讨。每种生长调节物质100μg溶于50％的丙酮中，每周1次处理于幼苗顶端，连续处理5周，并在第1次处理后24h，将幼苗水平放置。在实验期间，对照及GAs处理幼苗茎负向重性受到了促进，而Uni．-P处理则受到了抑制。GAs增加了茎上侧具有增厚细胞壁的胶质纤维细胞的数量。Uni．-P抑制了茎上侧的木材形成，但没有影响胶质纤维的分化。对照区木质部中纤维素与木质素的比值上侧大于下侧。该比值在上侧GA4处理区大于GA3处理区及对照区。该结果说明，胶质纤维的分化和木质部的增加对负向重性都是重要的；在该种中，赤霉素对于木质部细胞形成可能比对胶质纤维分化的影响更大。     

Stress and physiological,behavioral and performance patterns of children under varied air ion levels

The possibility that individual differences in reactivity to stressors are a major factor underlying discordant results reported for air ion studies prompted an investigation of response patterns in school children under both normal indoor air ion levels and moderately increased negative air ion levels (4000±500/cm³). It was hypothesized that the impact of stressors is reduced with high negative air ionization, and that resultant changes in stress effects would be differentially exhibited according to the children's normal degree of stimulus reactivity. A counter-balanced, replicative, withinssubject design was selected, and the subjects were 12 environmentally sensitive, 1st–4th grade school children. In addition to monitoring stress effects on activity level, attention span, concentration to task and conceptual performance, measures were also made of urinary 5-hydroxyindole acetic acid levels and skin resistance response (SRR) to determine if changes extended to the physiological state. The cold water test was used to add physical stress and enable calculations of Lacey's autonomic lability scores (ALS) as indicators of individual reactivity. The results show main effects for air ions on both physiological parameters, with 48% less change in %SRR (P<0.01) and 46% less change in urinary 5-HIAA levels (P<0.055) during negative air ions, indicating increased stress tolerance. Strong interactive effects for ALS x air ion condition appeared, with high and low ALS children reacting oppositely to negative air ions in measures of skin resistance level (P<0.01), wrist activity (P<0.01) and digit span backwards (P<0.004). Thus individual differences in autonomic reactivity and the presence or absence of stressors appear as critical elements for internal validity, and in preventing consequent skewed results from obscuring progress in air ion research.     

Diurnal changes in xylem pressure and mesophyll cell turgor pressure of the lianaTetrastigma voinierianum — the role of cell turgor in long-distance water transport: a comment

Summary The calculation of absolute-pressure values on the basis of measurements with differential-gauge pressure sensors, as described by Thürmer et al. (Protoplasma 206: 152–162, 1999), leads to discrepancies with the definition of absolute pressure when negative values are reached. From previous experiments with the xylem pressure probe we can conclude that the recorded pressure signal belongs not only to the xylem pressure, as stated by the authors, but also to the capillary pressure.     

Variability of pressure adaptation in deep sea bacteria

Isolations of pressure-adapted deep sea bacteria from depths of 1,400 to 5,100 m resulted in a variety of psychrophilic barotolerant and barophilic strains. Growth rates determined at different pressures indicated a gradual transition between the two types of pressure-adapted isolates. The presence of barotolerant bacteria in deep water, sustained by sinking particulate matter, causes the nonbarophilic response of natural populations, i.e., increased growth after decompression. With increasing pressure-adaptation in barophilic isolates the maximum growth rates at optimum pressures decrease. Thus, the observed general slow-down of microbial activity in the deep sea takes effect regardless of the common occurrence of psychrophilic and barophilic bacteria. The highest degree of barophilism was observed in isolates from nutrient-rich habitats such as intestinal tracts of deep sea animals or decaying carcasses. Detailed studies with an isolate, growing barophilically on a complex as well as a single-carbon-source medium, showed that (1) culturing at pressures lower than optimal for growth resulted in the formation of cell filaments, (2) growth was unaffected by repeated compression/decompression cycles and (3) no perceptible differences in the distribution of radiolabeled carbon from an amino acid mixture occurred in cells grown at, below and above the pressure optimal for growth.Dedicated to Professor Dr. Hans G. Schlegel on the occasion of his 60th birthday in recognition of his broad microbiological interests and in special appreciation of his lasting support for the Marine Microbiology Course at the Stazione Zoologica (Naples, Italy) now for almost 25 years Non-standard abbreviations. The traditional use of atm as a unit of pressure (=10 m of water column, =1.013 bar, =101.3 kN/m²) is retained here in view of the important relation between water depth and hydrostatic pressure in the present study. Due to the compression of seawater and the geographic variability of gravity, there is a progressive deviation of the actual pressure with depth amounting to +4.9 atm at 5,000 m and a latitude of 30°. EPC, cell counts obtained by epifluorescence microscopy. PY, peptone yeast extract medium     

On the statistical analysis of vegetation change: a wetland affected by water extraction and soil acidification

Abstract. A case study is presented on the statistical analysis and interpretation of vegetation change in a wetland subjected to water extraction and acidification, without precise information on the environmental changes. The vegetation is a Junco-Molinion grassland and the changes in vegetation are evaluated on the basis of relevés in 1977 and 1988 of 20 plots in a small nature reserve on moist oligotrophic, Pleistocene sands in the Netherlands. The changes are attributed to water extraction (since 1972) and soil acidification and the effect of the environmental changes on the vegetation is inferred from data on water depth and acidity collected in 1988. Many species typical of wetlands decreased in abundance, including rare species such as Parnassia palustris, Selinum carvifolia and Ophioglossum vulgatum. Some species increased, notably Anthoxanthum odoratum, Holcus lanatus and Plantago lanceolata. A significant decrease was found in the mean Ellenberg indicator values for moisture and acidity. The mean indicator value for nutrients did not change significantly. Multivariate analysis of the species data by Redundancy Analysis demonstrated the overall significance of the change in species composition between 1977 and 1988 (P < 0.01, Monte Carlo permutation). The spatial and temporal variation in the species data was displayed in ordination diagrams and interpreted in terms of water depth and pH. A simple model is developed to infer the change in water depth and pH from the relevé data and recent data on water depth and pH. Because the correlation between water depth and pH made a joint estimation of the changes useless, the change in pH was estimated for a series of likely changes in water depth. For the most likely change in water depth, significant acidification was inferred from the change in vegetation. The model is more generally applicable as a constrained calibration method.     

Pressure-induced change in proteins studied through chemical modifications

Pressure-induced change of two bovine proteins, α-lactalbumin (LA) and β-lactoglobulin (LG), was investigated at neutral pH by means of fluorescence and CD spectroscopy. The rate and the extent of modification was considerably increased by applying high pressure during the dansylation reaction of LG, while those for LA were only moderately affected. This difference was accounted for by the structural deformation of these proteins under high pressure. The fluorescence spectrum of these proteins measured under elevated pressure, as well as their fluorescence and CD spectra after the pressure release, indicated different responses towards pressure. The structural change of LA was practically reversible up to 400 MPa, whereas that of LG lost reversibility at 150 MPa or lower. Fluorescent measurement of dansylated (prepared at atmospheric pressure) proteins, especially the energy transfer from the intrinsic Trp residue to the dansyl group, showed that the protein structure was deformed by pressure and that the energy transfer facility of the two proteins was differently affected by high pressure, probably reflecting the degree of compactness of their pressure-perturbed structures