Chromosome aberrations induced in human lymphocytes by 8.7 MeV protons and 23.5 MeV helium-3 ions

This paper describes the irradiation of thin samples of blood with 8.7 MeV protons and 23.5 MeV helium-3 ions in the track segment mode. Chromosome aberrations in human lymphocytes have been scored. The relationship between dicentric yield and dose in Gy was Y = 0.044 D + 0.058 D2 for protons and Y = 0.394 D for helium ions. These results are compared with data from other laboratories using protons and an attempt is made to reconcile differences. An unexpected observation was that the ratio of the linear coefficients for helium ions and protons was about 9 whereas the ratio of the l.e.t. values was 4.5. This disagrees with current theory which predicts that the linear coefficients should be proportional to l.e.t. Possible sources of error in our experiments are discussed but do not adequately account for the discrepancies.     

Oscillatory Ca2+ signaling and its cellular function

It is well known that in the cells of many higher eukaryotic organisms Ca2+ ions are used as a signal messenger in the regulation of cellular functions. From recent studies with single cells it was suggested that the intracellular Ca2+ signal comprises repetitive and periodic Ca2+ spikes in a variety of cells. The mechanism by which intracellular Ca2+ oscillates and the biological significance of this oscillation are not well understood. It also remains to be determined how the Ca2+ signaling system sends a message into the cell, intermittently, to amplify the functional response. This review describes and integrates some recent views of oscillatory Ca2+ signaling.     

Binding and selectivity in L-type calcium channels: a mean spherical approximation

L-type calcium channels are Ca(2+) binding proteins of great biological importance. They generate an essential intracellular signal of living cells by allowing Ca(2+) ions to move across the lipid membrane into the cell, thereby selecting an ion that is in low extracellular abundance. Their mechanism of selection involves four carboxylate groups, containing eight oxygen ions, that belong to the side chains of the "EEEE" locus of the channel protein, a setting similar to that found in many Ca(2+)-chelating molecules. This study examines the hypothesis that selectivity in this locus is determined by mutual electrostatic screening and volume exclusion between ions and carboxylate oxygens of finite diameters. In this model, the eight half-charged oxygens of the tethered carboxylate groups of the protein are confined to a subvolume of the pore (the "filter"), but interact spontaneously with their mobile counterions as ions interact in concentrated bulk solutions. The mean spherical approximation (MSA) is used to predict ion-specific excess chemical potentials in the filter and baths. The theory is calibrated using a single experimental observation, concerning the apparent dissociation constant of Ca(2+) in the presence of a physiological concentration of NaCl. When ions are assigned their independently known crystal diameters and the carboxylate oxygens are constrained, e.g., to a volume of 0.375 nm(3) in an environment with an effective dielectric coefficient of 63.5, the hypothesized selectivity filter produces the shape of the calcium binding curves observed in experiment, and it predicts Ba(2+)/Ca(2+) and Na(+)/Li(+) competition, and Cl(-) exclusion as observed. The selectivities for Na(+), Ca(2+), Ba(2+), other alkali metal ions, and Cl(-) thus can be predicted by volume exclusion and electrostatic screening alone. Spontaneous coordination of ions and carboxylates can produce a wide range of Ca(2+) selectivities, depending on the volume density of carboxylate groups and the permittivity in the locus. A specific three-dimensional structure of atoms at the binding site is not needed to explain Ca(2+) selectivity.     

Transport ATPases: Structure,Motors, Mechanism and Medicine: A Brief Overview

Today we know there are four different types of ATPases that operate within biological membranes with the purpose of moving many different types of ions or molecules across these membranes. Some of these ions or molecules are transported into cells, some out of cells, and some in or out of organelles within cells. These ATPases span the biological world from bacteria to eukaryotic cells and have become most simply and commonly known as “transport ATPases.” The price that each cell type pays for transport work is counted in molecules of hydrolyzed ATP, a metabolic currency that is itself regenerated by a transport ATPase working in reverse, i.e., the ATP synthase. Four major classes of transport ATPases, the P, V, F, and ABC types are now known. In addition to being involved in many different types of biological/physiological processes, mutations in these proteins also account for a large number of diseases. The purpose of this introductory article to a mini-review series on transport ATPases is to provide the reader with a very brief and focused look at this important area of research that has an interesting history and bears significance to cell physiology, biochemistry, immunology, nanotechnology, and medicine, including drug discovery. The latter involves potential applications to a whole host of diseases ranging from cancer to those that affect bones (osteoporosis), ears (hearing), eyes (macromolecular degeneration), the heart (hypercholesterolemia/cardiac arrest,), immune system (immune deficiency disease), kidney (nephrotoxicity), lungs (cystic fibrosis), pancreas (diabetes and cystic fibrosis), skin (Darier disease), and stomach (ulcers).     

Genetic control of single lumen formation in the zebrafish gut

Most organs consist of networks of interconnected tubes that serve as conduits to transport fluid and cells and act as physiological barriers between compartments. Biological tubes are assembled through very diverse developmental processes that generate structures of different shapes and sizes. Nevertheless, all biological tubes invariably possess one single lumen. The mechanisms responsible for single lumen specification are not known. Here we show that zebrafish mutants for the MODY5 and familial GCKD gene tcf2 (also known as vhnf1) fail to specify a single lumen in their gut tube and instead develop multiple lumens. We show that Tcf2 controls single lumen formation by regulating claudin15 and Na+/K+-ATPase expression. Our in vivo and in vitro results indicate that Claudin15 functions in paracellular ion transport to specify single lumen formation. This work shows that single lumen formation is genetically controlled and appears to be driven by the accumulation of fluid.     

低能重离子在作物种胚内射程分布的研究技术

我国科学工作者率先将低能重离子束成功地应用于作物诱变育种,并建立了能量,质量和电荷三因子作用机制体系,但至今有关理论射程很短的低能重离子注入生物体后如何通过信息传递而诱发生物学效应的机理尚不完全清楚,低能重离子在作物种胚内的实际射程分布迄今仍是一个颇有争议的热点问题,而该项研究就直接触及低能离子束与生物组织细胞的原初作用机制,应用低能放射性束或具有可探测放射性的核反应产物,通过超薄切片和逐层分析测定,即可定量计算不同能量的低能离子束在作物种胚内的射程分布,本文还探讨了激光共聚焦扫描显微镜,原子力显微镜,X-射线能谱分析技术,单粒子微束技术和图像处理等技术途径在该项研究中的应用。     

Development of an ion microbeam system for irradiating single plant cell[s].

An ion microbeam system for irradiating single plant cells was developed to analyze exact biological effects of ion beams. Tobacco BY-2 protoplasts were used as a model of single plant cells. Protoplasts were cultured in thin agarose medium on a specially designed irradiation-vessel, which has a CR-39 nuclear track detector (a 100-micrometer thick sheet). The colony formation rate of unirradiated protoplasts was 22.7 +/- 6.7% (mean +/- SE of 3 different experiments) after a month of culture. Protoplasts were irradiated with programmed numbers of 18.3 MeV/u carbon ions that had been collimated by a 20-micrometer phi micro-aperture. After the irradiation, the positions within the protoplasts that were hit with ions were accurately determined by etching the CR-39 sheet in 13.4M KOH solution at 27 degrees centigrade for 9 h. The hit rate of the carbon ion microbeam, i.e., the percent of the ion particles that hit the protoplast that they were aimed at, was 56.9 +/- 2.4% (mean +/- SE of 7 different replications).     

Biophysical mechanism of radiation damage to mammalian cells by X- and gamma-rays

Published information on the reproductive death in mammalian cells irradiated by a wide range of X- and gamma-ray energies has been re-analysed to extract intrinsic efficiencies of damage for the secondary electrons in transient equilibrium. On a log-log plot, a linear dependence on the track average l.e.t. and the average specific primary ionization is found, indicating that either serves as a good quality parameter. The soft X-ray data are consistent with this conclusion. Upon comparison with data for fast heavy ion irradiations, the average specific primary ionization is shown to be applicable independently of radiation type whereas track average l.e.t. is not. Furthermore it is revealed that electrons are most damaging near the end of their range but their efficiency is only about 10-20 per cent of that of fast ions at the same quality, possibly due to the influence of multiple scatter on the electron penetration depth. It is deduced that, for the dose rates involved, the damage by electrons is predominantly by intra-track action and not inter-track action. The results are consistent with the suggestion that optimum damage occurs when the mean free path between ionizations is equivalent to the strand separation in the double-stranded DNA.     

Detergents induce raft-like domains budding and fission from giant unilamellar heterogeneous vesicles: a direct microscopy observation

The effect of detergents on giant unilamellar vesicles (GUVs) composed of phosphatidylcholine, sphingomyelin and cholesterol and containing liquid-ordered phase (l(o)) domains was investigated. Such domains have been used as models for the lipid rafts present in biological membranes. The studied detergents included lyso-phosphatidylcholine, the product of phospholipase A2 activity, as well as Triton X-100 and Brij 98, i.e. detergents used to isolate lipid rafts as DRMs. Local external injection of each of the three detergents at subsolubilizing amounts promoted exclusion of l(o) domains from the GUV as small vesicles. The budding and fission processes associated with this vesiculation were interpreted as due to two distinct effects of the detergent. In this framework, the budding is caused by the initial incorporation of the detergent in the outer membrane leaflet which increases the spontaneous curvature of the bilayer. The fission is related to the inverted-cone molecular shape of the detergent which stabilizes positively curved structures, e.g. pores involved in vesicle separation. On the other hand, we observed in GUVs neither domain formation nor domain coalescence to be induced by the addition of detergents. This supports the idea that isolation of DRM from biological membranes by detergent-induced extraction is not an artifact. It is also suggested that the physico-chemical mechanisms involved in l(o) domain budding and fission might play a role in rafts-dependant endocytosis in cells.     

Effect of different pion stopping volumes on the radiation response of mouse small intestine

This radiobiological investigation was based on measurements of crypt cell survival in mouse small intestine when the animals were exposed to 5 cm3 (spot), 350 cm3 and 3010 cm3 (the latter two were spot scans) pion stopping volumes generated by the multi-channel Piotron of the Swiss Institute for Nuclear Research. The experimental data obtained indicate that there was a decrease in biological effectiveness when the pion treatment volume was enlarged, irrespective of whether the pion dose was delivered in a single exposure or in four fractions. The r.b.e. values for pion beams relative to 200 kVp X-rays for the different experimental conditions used in this study are presented. The phenomenon of decreasing biological effectiveness with increasing pion stopping volume may be attributed to the following two factors: (1) when the pion stopping volume is increased there is a corresponding dilution of the high l.e.t. component of the beam; (2) the biological test system used may be sensitive to radiation dose rate which varied by a ratio of about 20 for the pion volumes used in this study.     

Effects of very low fluences of high-energy protons or iron ions on irradiated and bystander cells

In space, astronauts are exposed to radiation fields consisting of energetic protons and high atomic number, high-energy (HZE) particles at very low dose rates or fluences. Under these conditions, it is likely that, in addition to cells in an astronaut's body being traversed by ionizing radiation particles, unirradiated cells can also receive intercellular bystander signals from irradiated cells. Thus this study was designed to determine the dependence of DNA damage induction on dose at very low fluences of charged particles. Novel techniques to quantify particle fluence have been developed at the NASA Space Radiation Biology Laboratory (NSRL) at Brookhaven National Laboratory (BNL). The approach uses a large ionization chamber to visualize the radiation beam coupled with a scintillation counter to measure fluence. This development has allowed us to irradiate cells with 1 GeV/nucleon protons and iron ions at particle fluences as low as 200 particles/cm(2) and quantify biological responses. Our results show an increased fraction of cells with DNA damage in both the irradiated population and bystander cells sharing medium with irradiated cells after low fluences. The fraction of cells with damage, manifest as micronucleus formation and 53BP1 focus induction, is about 2-fold higher than background at doses as low as ～0.47 mGy iron ions (～0.02 iron ions/cell) or ～70 μGy protons (～2 protons/cell). In the irradiated population, irrespective of radiation type, the fraction of damaged cells is constant from the lowest damaging fluence to about 1 cGy, above which the fraction of damaged cells increases with dose. In the bystander population, the level of damage is the same as in the irradiated population up to 1 cGy, but it does not increase above that plateau level with increasing dose. The data suggest that at fluences of high-energy protons or iron ions less than about 5 cGy, the response in irradiated cell populations may be dominated by the bystander response.     

Famotidine, the new antiulcero-genic agent, a potent ligand for metal ions.

Potentiometric, polarographic, and spectroscopic results obtained for Cu2+ and Ni(2+)-famotidine systems clearly indicated that this anti-ulcerogenic drug is a very potent chelating agent able to coordinate cupric ion that was at pH below 2. This drug exhibits excellent histamine H2 receptor blocking effects and its effective coordination to metal ions may have significant biological implications. Famotidine is found to be a very effective ligand for Ni2+ ions also.     

Modeling the effects of low-LET cosmic rays on electronic components

The effects of cosmic radiation in single cells, organic tissues and electronics are a major concern for space exploration and manned missions. Standard heavy ions radiation tests employ ion cocktails with energy of the order of 10 MeV per nucleon and with a linear energy transfer ranging from a few MeV cm(2) mg(-1) to hundreds of MeV cm(2) mg(-1). In space, cosmic rays show significant fluxes at energies up to the order of GeV per nucleon. The present work aims at investigating single event damage due to low-, high- and very-high-energy ions. The European Space Agency reference single event upset monitor data are used to support the discussion. Finally, the effect of ionization induced directly by primary particles and ionization induced by recoils produced in an electronic device is investigated for different types of devices.     

Factors controlling the intracellular concentration of calcium and the spontaneous activity of the ureter

The role of the electrogenic Na(+)-Ca(2+)-exchange mechanism in regulating the spike activity of the ureter was studied. The ureter cells were shown to be capable of generating action potentials (AP) in sodium-free Krebs solution. The time during which the spikes are generated is in exponential dependence on the concentration of calcium ions in the medium, [Ca2+]o within 2.5 to 15 mmol/l. Simultaneously with the generation of the spikes, accumulation of calcium in the muscles is observed, proportional to the increase of [Ca2+]o. The addition of as little as 20 mmol/l Na+ or Li+ ions into the solution restores the prolonged electrical activity of the ureter. Under these conditions, the decrease of intracellular Ca2+ within 5 min was more than two times larger as compared with that in sodium-free medium. Upon substituting Ba2+ ions for Ca2+ ions in Krebs solution AP are generated within an interval which was the longer the higher the Ba2+ concentration in the medium. Li+ ions can replace Na+ ions in maintaining AP and in extruding calcium from the cell. It is supposed that the generation of the stable spike activity of the ureter depends on the functioning of Na(+)-Ca(2+)-exchange mechanism.     

Size-dependent ESS sex allocation in wind-pollinated cosexual plants: fecundity vs. stature effects

To theoretically investigate the single and compound effects of relative fecundity and relative stature of plants on size-dependent sex allocation (SDS) in wind-pollinated cosexual species, we developed a game model and analysed ESS sex allocation of large and small plants having totally or partially different reproductive resources and different pollen and seed dispersal areas in a population. We found that e.g. when both sized plants have large pollen dispersal areas relative to their seed dispersal areas, which plants are male-biased is largely determined by relative fecundity (t) and relative size of seed dispersal area (k) of the large plants to the small plants: If t >k, large plants tend to be more male-biased even if relative size of pollen dispersal area of large to small plants (l) is smaller than k. If t相似文献   

Effects of CO-2 and extracellular H+ iontophoresis on single cell activity in the cat brainstem.

The activity of single cells in deep regions of the medulla oblongata was observed both during CO2 inhalation and during the extracellular iontophoresis of hydrogen ions in peripheral chemoreceptor-denervated cats. All 53 neurons that fired in synchrony with some part of the ventilatory cycle showed increased firing during CO2 inhalation; yet none responded in a graded fashion to the extracellular application of hydrogen ions. Seventy-one of the 74 nonperiodic cells studied showed no response to CO2 inhalation. Of the 3 nonperiodic cells that did respond to CO2, 2 also responded in a graded fashion to the extracellular iontophoresis of hydrogen ions. It is concluded that the cell bodies of medullary neurons with respiratory periodicity are relatively insensitive to hydrogen ions. Further the paucity of hydrogen ion-sensitive cells found in deep areas of the medulla does not support the notion that medullary hydrogen ion chemoreception is largely achieved by structures located deep in the lower brainstem.     

Characteristics of a microbial assay for the detection of halogenated hydrocarbons using cells of an actinomycete-like organism as a biological component

Cells of an Actinomycete-like bacterium, strain GJ70, with the ability to degrade several haloalkanes were used as a biological component in a discontinuous microbial bioassay for the detection of 1,3-dichloropropene and 1,2-dibromoethane in water. The cells were entrapped in different matrices such as calcium alginate, carrageenan, chitosan, polyacrylamide-hydrazide and chitosan-carboxy-methyl cellulose; the specific dehalogenating activity of the immobilized cells to a stirred sample solution and by the use of an ion selective electrode (ISE) for the quantification of enzymatically released halogen ions, the concentration of halogenated hydrocarbons could be estimated by determining the change of electrode potential within a period of 5 min. The detection limits for 1,3-dichloropropene and 1,2-dibromoethane were below 100 μg/l and 25 μg/l, respectively; the relative standard deviation was < 10%. In addition, several chlorinated and brominated hgydrocarbons were converted by the bacterial cells at a reduced rate e.g. 1, 2-dibromopropane, 1-bromoethane, 1,5-dichloropentane, etc. Moreover, temperatures of between 20 and 40%C did not affect the enzymatic activity of the cells, and a pH of between at 5 and 9 had little influence. Several organic substances and non-metabolizable compounds did not affect the conversion, whereas some heavy metal ions acted as inhibitors.     

Study on effect of diluent osmotic pressure on yeast cell strength and cell size using a novel micromanipulation technique

A new micromanipulation technique which has previously been used to measure the mechanical properties of single animal cells has now been applied to yeast cells. In this study this technique was used to measure yeast cell strength and cell size across a 2l batch fermentation. Alternatively the cell size could also be determined using a Coulter counter while cell measurement was diluted with a conducting fluid (Isoton II). For the cell strength, it was found that the osmotic pressure of diluents did affect cell strength. However, it was also found that there was no significant effect of osmotic pressure of diluents on cell size whether a Coulter counter or micromanipulation was used for measurement. Micromanipulation has been shown to be a powerful technique for measuring the mechanical properties of yeast cells and it will be very useful for studying their behaviour in cell disruption equipment, e.g. high-pressure homogenizers.     

Activation of mitochondrial glycerol 3-phosphate dehydrogenase by cadmium ions

Mitochondrial glycerol 3-phosphate dehydrogenase (EC 1.1.2.1.) requires Ca2+ ions for its activity. Cadmium ions also have activatory effect on the enzyme. They activate the glycerol 3-phosphate dehydrogenase in a very narrow concentration range (1-2 mmol/l). As contrasted with calcium, strong inhibitory effect occurred at higher concentrations (3-4 mmol/l). The inhibition induced by cadmium ions was completely reversible by washing of the mitochondria.