Redistribution of stress due to a circular hole in a nonlinear anisotropic membrane

Many clinical procedures introduce holes into thin tissues that are typically under multiaxial stresses and finite strains. Such incisions change the stresses and strains from their homeostatic values, which may induce cells to alter their orientation and cytoskeletal organization as well as to migrate, proliferate, change their synthesis of matrix, or even to enter the cell death cycle. To correlate such changes in cellular activity with changes in the mechanics, we need solutions for the native and the perturbed boundary value problems. Such problems will often be complex and require a finite element solution; weak solutions should be evaluated independently, however, at least for special cases. Herein, we present a numerical solution of the governing nonlinear ordinary differential equation for the special case of stress redistribution due to the introduction of a circular hole into a finitely deformed, Fung-type, circular membrane that exhibits a cylindrical orthotropy. Among other results, we show that the anisotropy plays an increasingly greater role as the size of the hole becomes smaller, which is of course a goal of minimally invasive procedures.     

The central reactions of nitrogen fixation

Summary An outline is given of the general enzymology of the nitrogen-fixing enzyme complex, nitrogenase and thermodynamic considerations of the function of ATP in the reaction are introduced. Some of the implications of these features of the central reactions of nitrogen fixation are discussed in relation to the role of the process in biological productivity.     

Predicting and mapping the spatial distribution of Chamaecyparis formosensis in central Taiwan in a GIS with species distribution models

Analyzing the relationship between species and environment is always a focal question of ecological research. In recent years species distribution models (SDMs) has been widely used to predict the spatial distribution of species. SDMs are numerical tools that combine observations and species occurrence or abundance with environmental variables to predict the spatial distribution of species across landscapes, sometimes requiring extrapolation in space and time. Chamaecyparis formosensis (Taiwan red cypress, TRCs) is a coniferous species endemic to Taiwan, where it natural grows in the central mountains at moderate to high altitudes of 800–2800 m, and most stands in the range of 1500–2150 m. It is threatened by habitat loss and over-cutting for its valuable timber. To preserve TRCs species and achieve sustainable use of biological resources, we choose TRCs as a target for the study to predict its distribution in central Taiwan.The pure forests of TRCs in the study area were mainly located in Pachsienshan (P), Shouchentashan (S) and Baigou Mountain (B) in central Taiwan, and the distribution data were originally obtained by The Third Survey of Forest Resources and Land Use in Taiwan. Elevation, slope, aspect, and three vegetation indices were derived from both SPOT-5 satellite images and DEM. GIS technique was used to overlay those factors. Discriminant analysis (DA), decision tree (DT) and maximum entropy (MAXENT), three commonly used SDMs, were applied based on above-mentioned six variables to predict the suitable habitat of TRCs, and to evaluate which the best model is in terms of accuracy and efficiency. Three experiment designs (ED1, ED2 and ED3) with different combinations of samples were used for model building and validation. The 200 target samples were collected from the site P–B, B–S and P–S for model building under ED1, ED2 and ED3 respectively, while the 100 samples were collected from the site S, P and B for model validation. All experiment designs had same 1350 background samples. The results showed that the overall accuracy and kappa coefficient of DT (96%, 0.88) was higher than that of MAXENT (91%, 0.70), and their accuracies were better than that of DA (84%, 0.58). All the three models were highly efficient in implementation of model construction and evaluation, while the DT model was difficult for generating the entire predicted map of potential habitat due to its complex conditional sentence. Vegetation indices derived from SPOT-5 satellite images could not improve model accuracy because of its insufficiency of spectral resolution and spatial resolution. High spatial resolution and spectral resolution remotely sensed imagery should be used in our future research to improve model performance and reliability.     

Photoselection and circular dichroism in the purple membrane.

The transient dichroic ratio D = delta A parallel/delta A perpendicular has been measured in the visible absorption region of bacteriorhodopsin in purple membrane by a flash photolysis method. D is found to be wavelength independent throughout the visible absorption band, and reaches a maximum value of 2.75 +/- 0.15 on reduction of the excitation intensity. This value is close to that expected for a single nondegenerate transition dipole moment and is incompatible with the strong exciton coupling model used to explain circular dichroism (CD) spectrum of purple membrane. A time-dependent analysis of the exciton interaction and consideration of the coupling strength suggests an explanation of these observations. It is concluded that excitation interaction between retinals in purple membrane is of the weak or very weak type defined by Förster.     

On stability of circular hole in membrane bilayer.

It was observed recently that nonionic surfactant octaethyleneglycol dodecylether (Cl2E8) decreases threshold for irreversible electroporation in membrane bilayers. In accordance, it is shown theoretically in this work that anisotropic C12E8 membrane inclusions may stabilize circular hole in a flat membrane segment.     

Extracellular recording with transistors and the distribution of ionic conductances in a cell membrane

Intracellular voltage transients of cultured cells are recorded by transistors and other planar electrodes as local extracellular voltages. The theoretical relationship between extra- and intracellular voltage is investigated with a two-compartment circuit using the approximation of a fast, weak and small cell-silicon junction. It is shown that extracellular recording relies on the difference of specific ionic conductances in the attached and free regions of the cell membrane. The result rationalizes various observations with neuron transistors. It guides the optimization of extracellular recording and the development of cell-based chemical sensors. Received: 13 August 1998 / Revised version: 9 October 1998 / Accepted: 9 October 1998     

Identification of a synaptic vesicle-specific membrane protein with a wide distribution in neuronal and neurosecretory tissue

Two different monoclonal antibodies, characterized initially as binding synaptic terminal regions of rat brain, bind a 65,000-dalton protein, which is exposed on the outer surface of brain synaptic vesicles. Immunocytochemical experiments at the electron microscope level demonstrate that these antibodies bind the vesicles in many different types of nerve terminals. The antibodies have been used successfully to purify synaptic vesicles from crude brain homogenates by immunoprecipitation onto the surface of polyacrylamide beads. The profiles of the structures precipitated by these beads are almost exclusively vesicular, confirming the vesicle-specificity of the antibodies. In SDS gels, the antibodies bind a single protein of 65,000 daltons. The two antibodies are not identical, but compete for binding sites on this protein. Immune competition experiments also demonstrate that the antigenic components on the 65,000-dalton protein are widely distributed in neuronal and neural secretory tissues. Detectable antigen is not found in uninnervated tissue--blood cells and extrajunctional muscle. Low levels are found in nonneural secretory tissues; it is not certain whether this reflects the presence of low amounts of the antigen on all the exocytotic vesicles in these tissues or whether the antigen is found only in neuronal fibers within these tissues. The molecular weight and at least two antigenic determinants of the 65,000-dalton protein are highly conserved throughout vertebrate phylogeny. The two antibodies recognize a 65,000-dalton protein present in shark, amphibia, birds, and mammals. The highly conserved nature of the determinants on this protein and their specific localization on secretory vesicles of many different types suggest that this protein may be essential for the normal function of neuronal secretory vesicles.     

Acetyl CoA,a central intermediate of autotrophic CO2 fixation in Methanobacterium thermoautotrophicum

The pathway of autotrophic CO₂ fixation in Methanobacterium thermoautotrophicum has been investigated by long term labelling of the organism with isotopic acetate and pyruvate while exponentially growing on H₂ plus CO₂. Maximally 2% of the cell carbon were derived from exogeneous tracer, 98% were synthesized from CO₂. Since growth was obviously autotrophic the labelled compounds functioned as tracers of the cellular acetyl CoA and pyruvate pool during cell carbon synthesis from CO₂. M. thermoautotrophicum growing in presence of U-¹⁴C acetate incorporated ¹⁴C into cell compounds derived from acetyl CoA (N-acetyl groups) as well as into compounds derived from pyruvate (alanine), oxaloacetate (aspartate), -ketoglutarate (glutamate), hexosephosphates (galactosamine), and pentosephosphates (ribose). The specific radioactities of N-acetylgroups and of the three amino acids were identical. The hexosamine exhibited a two times higher specific radioactivity, and the pentose a 1.6 times higher specific radioactivity than e.g. alanine. M. thermoautotrophicum growing in presence of 3-¹⁴C pyruvate, however, did not incorporate ¹⁴C into cell compounds directly derived from acetyl CoA. Those compounds derived from pyruvate, dicarboxylic acids and hexosephosphates became labelled. The specific radioactivities of alanine, aspartate and glutamate were identical; the hexosamine had a specific radioactivity twice as high as e.g. alanine.The finding that pyruvate was not incorporated into compounds derived from acetyl CoA, whereas acetate was incorporated into derivatives of acetyl CoA and pyruvate in a 1:1 ratio demonstrates that pyruvate is synthesized by reductive carboxylation of acetyl CoA. The data further provide evidence that in this autotrophic CO₂ fixation pathway hexosephosphates and pentosephosphates are synthesized from CO₂ via acetyl CoA and pyruvate.     

Stress sensitive glucose oxidase-nylon membrane

Summary Glucose oxidase (GOD) was immobilized on a nylon membrane. No activity of the GOD-nylon membrane was observed under normal conditions but it appeared when the membrane was mechanically stretched. A linear relationship was observed between the stress strength and the GOD activity of the membrane. The appearance of the GOD activity of the membrane with stress was reproducible and the membrane could be stored for at least 2 months. Therefore, the GOD-nylon membrane can be called a stress sensitive membrane.     

SNARE interactions in membrane trafficking: a perspective from mammalian central synapses

SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) are a large family of proteins that are present on all organelles involved in intracellular vesicle trafficking and secretion. The interaction of complementary SNAREs found on opposing membranes presents an attractive lock-and-key mechanism, which may underlie the specificity of vesicle trafficking. Moreover, formation of the tight complex between a vesicle membrane SNARE and corresponding target membrane SNAREs could drive membrane fusion. In synapses, this tight complex, also referred to as the synaptic core complex, is essential for neurotransmitter release. However, recent observations in knockout mice lacking major synaptic SNAREs challenge the prevailing notion on the executive role of these proteins in fusion and open up several questions about their exact role(s) in neurotransmitter release. Persistence of a form of regulated neurotransmitter release in these mutant mice also raises the possibility that other cognate or non-cognate SNAREs may partially compensate for the loss of a particular SNARE. Future analysis of SNARE function in central synapses will also have implications for the role of these molecules in other vesicle trafficking events such as endocytosis and vesicle replenishment. Such analysis can provide a molecular basis for synaptic processes including certain forms of short-term synaptic plasticity.     

Analyses of circular dichroism spectra of membrane proteins

Circular dichroism (CD) spectroscopy is a valuable technique for the determination of protein secondary structures. Many linear and nonlinear algorithms have been developed for the empirical analysis of CD data, using reference databases derived from proteins of known structures. To date, the reference databases used by the various algorithms have all been derived from the spectra of soluble proteins. When applied to the analysis of soluble protein spectra, these methods generally produce calculated secondary structures that correspond well with crystallographic structures. In this study, however, it was shown that when applied to membrane protein spectra, the resulting calculations produce considerably poorer results. One source of this discrepancy may be the altered spectral peak positions (wavelength shifts) of membrane proteins due to the different dielectric of the membrane environment relative to that of water. These results have important consequences for studies that seek to use the existing soluble protein reference databases for the analyses of membrane proteins.     

Plasma membrane of Synechocystis PCC 6803: a heterogeneous distribution of membrane proteins

Proteomic studies carried out previously on the plasma membrane of Synechocystis have identified several peripheral and integral proteins. The distribution of these proteins along the membrane still remains obscure. In this study, the distribution of proteins along the plasma membrane of Synechocystis was carried out using subfractions, the right-side-out (RSO) and inside-out (ISO) vesicles, fractionated from a pure and specific fraction of the plasma membrane. These subfractions were analyzed and quantified for several proteins by immunoblotting. It was found that the ISO fraction contained higher quantities of preD1, D1 and PsaD, the integral proteins of photosystem I and II known to be present also in the plasma membrane. Lower amounts of peripheral vesicle inducing protein Vipp1 and nitrate/nitrite binding protein NrtA were present in the ISO compared to the RSO fraction. On the contrary, the distribution of two integral transporter proteins, SbtA and PxcA, was found equal in both fractions. Our studies clearly establish that the plasma membrane of Synechocystis has a heterogeneous composition with respect to protein distribution. The accumulation of photosynthesis-associated proteins in the ISO fraction provides evidence that the discrete regions of the plasma membrane harbor sites for biogenesis of photosystems.     

Factors associated with woody alien species distribution in a newly invaded mountain system of central Argentina

To help determine the major factors associated with alien plant in a newly invaded mountain range; we analyzed the distribution patterns of woody alien species along the altitudinal gradient of the Córdoba mountains, in relation to biotic, abiotic and anthropogenic factors. We selected 303 plots using a Geographic Information System (GIS) covering all the variability of these factors. In the field we registered woody alien occurrence in these 303 and in 303 additional neighbor plots. We used 12 biotic, abiotic and anthropogenic variables from the GIS to predict the probability of presence of alien species through a logistic model. Then, we analyzed if neighbor alien occurrences could explain some additional variance. We created a probability map with 4 categories of alien occurrence which was then validated by new field sampling. Occurrence of woody aliens was highest in the eastern slope (with longer history of species introduction), at low altitudes, near sources of propagule pressure (human settlements, roads and neighbor sites with established alien plants), and associated to sheltered topographies. In the upper belt of the Córdoba Mountains woody invasion is incipient and thus in a transient stage. Accordingly, propagule pressure seems to be the major factor at play, while the relevance of disturbance and biotic interactions is less clear.     

Simultaneous fixation and production of catecholamine fluorescence in central nervous tissue by perfusion with aldehydes

Synopsis Perfusion with a mixture of formaldehyde (4%) and glutaraldehyde (0.5%) is shown both to fix central nervous tissue and to produce, with no further treatment, a fluorescence histochemical localization of catecholamines. After perfusion, serial sections can be readily taken through the whole brain with a Vibratome. Landmarks which are apparent at low power with white-light illumination can be seen when the sections are viewed in the fluorescence microscope. Catecholamine-containing nerve cell bodies, varicose axon terminals and non-varicose nerve fibres appear brightly fluorescent and well localized. The method has been applied to rats, guinea-pigs and rabbits and is ideally suited to the accurate mapping of central catecholamine neurons and their processes.