Electrorotation of Erythrocytes Treated with Dipicrylamine: Mobile Charges within the Membrane Show their ``Signature' in Rotational Spectra

In this study, electrorotation spectra of individual cells (that is, frequency dependence of cell rotation speed) have been proved to yield information not only about the passive electric properties of cell constituents, but also about the presence of mobile charges within the plasma membrane being part of ion carrier transport systems. Experiments on human erythrocytes pretreated with the lipophilic anion dipicrylamine (DPA) gave convincing evidence that these artificial mobile charges adsorbed to the plasma membrane contributed significantly to the rotational spectrum at relatively low conductivity of the external medium (2–5 mS m⁻¹). Theoretical integration of the mobile charge concept into the single-shell model (viewing the cell as a homogenous sphere surrounded by a membrane) led to a set of equations which predicted electrorotational behavior of DPA-treated cells in dependence on medium conductivity. The quantitative data on the partition and the transmembrane translocation rate of the DPA anion extracted from the experimental rotational spectra agreed well with the corresponding literature values. Received: 14 February 1996/Revised: 29 May 1996     

Growth and crown efficiency of height repressed lodgepole pine; are suppressed trees more efficient?

Leaf area, crown projection area and growth over the last 5 years were measured to assess growth efficiency (GE) and crown efficiency (CE) of dominant (D), codominant (CD) and suppressed (SP) trees growing in height-repressed (P sites) and normally developing (M sites) lodgepole pine stands. Leaf area index (LAI), hydraulic characteristics, and needle nutrient concentrations were also measured. Volume growth of P site trees between 1994 and 1999 was 46% that of M site trees. Volume growth was closely associated with both hydraulic supply capacity (Q*) and leaf area. Height repression was not associated with lower GE, but P site trees had CE that was 24.5% lower than M site trees. Average GE of D and CD trees was 28% lower than that of SP trees, while mean CE for the D trees was 46% greater than that of CD, and 80% greater than for SP trees. Between M and P sites, canopy LAI and Q* per unit leaf area did not differ. Needle nitrogen (N) concentrations of M site trees were 7.6% greater than for P site trees. SP tree needles had the highest concentration of N and phosphorus. The nutrient advantage enjoyed by SP trees presumably allowed them to maintain higher GE for a given Q*/A_l. The fastest growing trees were the large D and CD trees from M sites. As LAI did not differ between sites, height repression on P sites may be a result of total leaf area being distributed among too many small trees.     

Reversible Electropermeabilization of Mammalian Cells by High-Intensity, Ultra-Short Pulses of Submicrosecond Duration

Mouse myeloma cells were electropermeabilized by single square-wave electric pulses with amplitudes of up to ∼150 kV/cm and durations of 10–100 nsec. The effects of the field intensity, pulse duration and medium conductivity on cell viability and field-induced uptake of molecules were analyzed by quantitative flow cytometry using the membrane-impermeable fluorescent dye propidium iodide as indicator molecule. Despite the extremely large field strengths, the majority of cells survived the exposure to ultra-short field pulses. The electrically induced dye uptake increased markedly with decreasing conductivity of the suspending medium. We assigned this phenomenon to the transient electrodeformation (stretching) force that assumes its maximum value if cells are suspended in low-conductivity media, i.e., if the external conductivity σ_e is smaller than that of the cytosol σ_i. The stretching force vanishes when σ_e is equal to or larger than σ_i. Due to their capability of delivering extremely large electric fields, the pulse power systems used here appear to be a promising tool for the electropermeabilization of very small cells and vesicles (including intracellular organelles, liposomes, etc.). Received: 15 May 2001/Revised: 20 July 2001     

Microbial community composition and hydrochemistry of underexplored geothermal waters in Croatia

In Croatia, a variety of geothermal springs with a wide temperature range and varied hydrochemical conditions exist, and they may harbor different niches for the distribution of microbial communities. In this study, 19 different sites, mainly located in central and eastern Croatia, were selected for primary characterization of spring hydrochemistry and microbial community composition. Using 16S rRNA gene amplicon sequencing, it was found that the bacterial communities that dominated most geothermal waters were related to Proteobacteria and Campylobacteria, while most archaeal sequences were related to Crenarchaeota. At the genus level, the prokaryotic community was highly site-specific and was often dominated by a single genus, including sites dominated by Hydrogenophilus, Sulfuricurvum, Sulfurovum, Thiofaba and Nitrospira, while the most abundant archaeal genera were affiliated to the ammonia-oxidizing archaea, Candidatus Nitrosotenuis and Candidatus Nitrososphaera. Whereas the microbial communities were overall highly location-specific, temperature, pH, ammonia, nitrate, total nitrogen, sulfate and hydrogen sulfide, as well as dissolved organic and inorganic carbon, were the abiotic factors that significantly affected microbial community composition. Furthermore, an aquifer-type effect was observed in the community composition, but there was no pronounced seasonal variability for geothermal spring communities (i.e. the community structure was mainly stable during the three seasons sampled). These results surprisingly pointed to stable and geographically unique microbial communities that were adapted to different geothermal water environments throughout Croatia. Knowing which microbial communities are present in these extreme habitats is essential for future research. They will allow us to explore further the microbial metabolisms prevailing at these geothermal sites that have high potential for biotechnological uses, as well as the establishment of the links between microbial community structure and the physicochemical environment of geothermal waters.     

山珍补益膏的营养研究

山珍补益膏含有较高含量的各种必需氨基酸、微量元素和维生素。     

Water beetle tolerance to salinity and anionic composition and its relationship to habitat occupancy

Water salinity and ionic composition are among the main environmental variables that constrain the fundamental niches of aquatic species, and accordingly, physiological tolerance to these factors constitutes a crucial part of the evolution, ecology, and biogeography of these organisms. The present study experimentally estimated the fundamental saline and anionic niches of adults of two pairs of congeneric saline beetle species that differ in habitat preference (lotic and lentic) in order to test the habitat constraint hypothesis. Osmotic and anionic realised niches were also estimated based on the field occurrences of adult beetle species using Outlying Mean Index analysis and their relationship with experimental tolerances. In the laboratory, all of the studied species showed a threshold response to increased salinity, displaying high survival times when exposed to low and intermediate conductivity levels. These results suggest that these species are not strictly halophilic, but that they are able to regulate both hyperosmotically and hypoosmotically. Anionic water composition had a significant effect on salinity tolerance at conductivity levels near their upper tolerance limits, with decreased species survival at elevated sulphate concentrations. Species occupying lentic habitats demonstrated higher salinity tolerance than their lotic congeners in agreement with the habitat constraint hypothesis. As expected, realised salinity niches were narrower than fundamental niches and corresponded to conditions near the upper tolerance limits of the species. These species are uncommon on freshwater-low conductivity habitats despite the fact that these conditions might be physiologically suitable for the adult life stage. Other factors, such as biotic interactions, could prevent their establishment at low salinities. Differences in the realised anionic niches of congeneric species could be partially explained by the varying habitat availability in the study area. Combining the experimental estimation of fundamental niches with realised field data niche estimates is a powerful method for understanding the main factors constraining species’ distribution at multiple scales, which is a key issue when predicting species’ ability to cope with global change.     

Electroporation in dense cell suspension—Theoretical and experimental analysis of ion diffusion and cell permeabilization

Electroporation is a process where increased permeability of cells exposed to an electric field is observed. It is used in many biomedical applications including electrogene transfection and electrochemotherapy. Although the increased permeability of the membrane is believed to be the result of pores due to an induced transmembrane voltage U_m, the exact molecular mechanisms are not fully explained.