Role of the plasma interaction with magnetic field in the “missing power” problem

It is shown that, at rapid changes of the heating power, the magnetically confined equilibrium plasma almost completely absorbs the injected energy, so that its only small part goes to the magnetic field. The result is obtained within the standard MHD theory with use of exact consequences of the force-balance equations in toroidal geometry. It is assumed that, when heated, the plasma evolves from one equilibrium state to another with the magnetic field frozen-in. Another constraint is the conservation of the toroidal magnetic flux in the plasma-wall vacuum gap. It is shown that the plasma interaction with magnetic field (which is traditionally neglected in the analysis of heat transport in tokamaks and stellarators) is the natural mechanism of fast redistribution of energy in the plasma, observed in some experiments in these devices at switchon of powerful heat sources.     

Investigations of the turnover of the putative Cellulose-synthesizing particle “rosettes” within the plasma membrane ofFunaria hygrometrica protonema cells

Summary YoungFunaria protonemata were treated with Monensin (10^–6 M) and Cytochalasin (CB) (2×10^–5 M). The influence of the inhibitors on a) elongation growth, b) cell fine structure and c) particle rosettes within the plasma membrane after freeze fracture was observed. Monensin stopped cell growth, caused swelling of the mitochondria and plastids and inhibited the secretory activity of the Golgi apparatus within about 15 minutes. The number of rosettes in the PF of the plasma membrane was distinctly reduced after 4–5 minutes and decreased further to only very few after 30 minutes. The tip to base gradient in distribution was maintained for a long time. The effects were reversible, regeneration occurred within 3 hours. CB treatment showed no effect on elongation growth and cell fine structure. The number of rosettes, however, was strongly reduced within 3 minutes exposure time and their distribution was nearly uniform then. Number and tip to base gradient increased again after 6 minutes intoxication. The results are discussed in regard to the turn over of the rosettes.Abbreviations CB Cytochalasin B - PF protoplasmic fracture face - F-vesicle flat vesicle - F-Actin filamentous actin - G-Ac-tin globular actin     

Are the conformational dynamics and the ligand binding properties of myoglobin affected by exposure to microwave radiation?

The global uptake of mobile communication emphasizes the question about possible adverse consequences of the exposure to low-level radiofrequency radiation from mobile phones on human health as result of so-called "non-thermal effects". In order to state safety guidelines it seems appropriate to start by excluding, if possible, non-specific effects on structural and dynamic properties of fundamental biomolecules such as proteins. Proteins are flexible polyelectrolytes; thus, they are susceptible, in principle, to the action of electromagnetic fields. In this article, we investigated the effects of microwaves on structural and functional properties of Tunnus tynnus myoglobin at 1.95 GHz, a frequency used by new wireless microwave communication systems. The protein solution was exposed for 2.5 h to 51 mW/g SAR (specific absorption rate) level. Measurements of absorption spectroscopy, circular dichroism and fluorescence emission decay in the frequency domain do not exhibit any influence of the radiation on the native structural state of protein macromolecules.     

Selective block by α-dendrotoxin of the K+ inward rectifier at the Vicia guard cell plasma membrane

The efficacy and mechanism of -dendrotoxin (DTX) block of K⁺ channel currents in Vicia stomatal guard cells was examined. Currents carried by inward- and outward-rectifying K⁺ channels were determined under voltage clamp in intact guard cells, and block was characterized as a function of DTX and external K⁺ (K⁺) concentrations. Added to the bath, 0.1-30 nM DTX blocked the inward-rectifying K⁺ current (I_K,in), but was ineffective in blocking current through the outward-rectifying K⁺ channels (I_K,out) even at concentrations of 30 nM. DTX block was independent of clamp voltage and had no significant effect on the voltage-dependent kinetics for I_K,in, neither altering its activation at voltages negative of –120 mV nor its deactivation at more positive voltages. No evidence was found for a use dependence to DTX action. Block of I_K,in followed a simple titration function with an apparent K_1/2 for block of 2.2 nM in 3 mm K _o ⁺ . However, DTX block was dependent on the external K⁺ concentration. Raising K⁺ from 3 to 30 mm slowed block and resulted in a 60–70% reduction in its efficacy (apparent K _i = 10 mm in 10 nm DTX). The effect of K⁺ in protecting I _K,in was competitive with DTX and specific for permeant cations. A joint analysis of I_K,in block with DTX and K⁺ concentration was consistent with a single class of binding sites with a K _d for DTX of 240 pm. A K _d of 410 m for extracellular K⁺ was also indicated. These results complement previous studies implicating a binding site requiring extracellular K⁺ (K_1/2 1 mm) for I_K,in activation; they parallel features of K⁺ channel block by DTX and related peptide toxins in many animal cells, demonstrating the sensitivity of plant plasma membrane K⁺ channels to nanomolar toxin concentrations under physiological conditions; the data also highlight one main difference: in the guard cells, DTX action appears specific to the K⁺ inward rectifier.We thank J.O. Dolly (Imperial, London) and S.M. Jarvis (University of Kent, Canterbury) for several helpful discussions. This work was supported by SERC grant GR/H07696 and was aided by equipment grants from the Gatsby Foundation, the Royal Society and the University of London Central Research Fund. G.O. was supported by an Ausbildungsstipendium (OB 85/1-1) from the Deutsche Forschungsgemeinschaft. F.A. holds a Sainsbury Studentship.     

Displacement of “pseudoanomeric” hydroxyl groups by using the diethyl azodicarboxylate-triphenylphosphine system

1d-(1,2,4/3)-2,3,4-Tri-O-benzyl-5-(trityloxymethyl)-5-cyclohexene-1,2,3,4-tetrol (5a) and its 1l-(1,3/2,4) isomer (5b) were prepared from d-glucose, and they underwent ready mutual interconversion through an S_N2 procedure employing a benzoic acid-diethyl azodicarboxylate-triphenylphosphine system and subsequent basic hydrolysis. Azido, phthalimido, and even more complex nucleophile groups could similarly also substitute the allylic hydroxyl groups of 5a and 5b by using the same system, with a few different results between 5a and 5b.     

The “double grafted tumor system”, proposed to find effector cells in the analyses of antitumor effect of BRMs

The antitumor effects of three biological response modifiers (BRMs; PSK, IFN A/D and OK432) and two chemotherapeutics (Mitomycin C and Neocarzinostatin) in a new experimental mouse model, the double grafted tumor system, were evaluated. BALB/c mice received simultaneous inoculations of Meth A fibrosarcoma cells on right flank (1 × 10⁶ cells) and left flank (2 × 10⁵ cells) on day 0, and drugs were given intratumorally into the right-flank tumor on day 3. The growth of the left-flank tumor was the real target for the evaluation of a given drug after 21 days. All tested five agents successfully cured the drug-injected right tumor with a pre-determined optimum dose. In addition, PSK, OK432, IFN A/D and MMC among the five, inhibited the left-flank tumor, whereas no inhibition was observed when treated with NCS. To understand the mechanism by which the antitumor effect of the above four agents is able to influence the growth of tumor on the other side, tumor cells (2 × 10⁵ cells) inoculated only into the left flank were treated with drugs given subcutaneously to the right flank (single tumor system). Among the four, MMC exhibited an effect similar to that obtained in the double tumor system, and IFN A/D showed a less pronounced but still definite antitumor effect. However, PSK and OK432 failed to express anti-tumor effect in the single tumor system. These results obtained with PSK, OK432 and IFNA/D suggest that the effect of the drug on the left-tumor may be mediated by certain effector cells, which are specifically induced by injection of the drug, in the right-tumor tissues. When effector cell analysis was conducted with spleen cells obtained after PSK treatment by means of intratumoral adoptive transfer into 3-day Meth A bearing recipients, these cells were shown to be Lyt-1⁺2^–-T and L3T4⁺-T cell.     

First step toward the “fingerprinting” of brain tumors based on synchrotron radiation X-ray fluorescence and multiple discriminant analysis

Synchrotron-radiation-based X-ray fluorescence was applied to the elemental microimaging of neoplastic tissues in cases of various types of brain tumors. The following cases were studied: glioblastoma multiforme, gemistocytic astrocytoma, oligodendroglioma, anaplastic oligodendroglioma, ganglioglioma, fibrillary astrocytoma, and atypical transitional meningioma. Apart from neoplastic tissue, the analysis included areas of tissue apparently without malignant infiltration. The masses per unit area of P, S, Cl, K, Ca, Fe, Cu, Zn, Br, and Rb were used to construct a diagnostic classifier for brain tumors using multiple discriminant analysis. It was found that S, Cl, Cu, Fe, K, Br, and Zn are the most significant elements in the general discrimination of tumor type. The highest similarity in elemental composition was between atypical transitional meningioma and fibrillary astrocytoma. The smallest differentiation was between glioblastoma multiforme and oligodendroglioma. The mean percentage of correct classifications, estimated according to the a posteriori probabilities procedure, was 99.9%, whereas the mean prediction ability of 87.6% was achieved for ten new cases excluded previously from the model construction. The results showed that multiple discriminant analysis based on elemental composition of tissue may be a potentially valuable method assisting differentiation and/or classification of brain tumors.     

Synopsis of a new “polyphyletic-polychronic-polytopic” system of the angiosperms

This work is continuation of two papers published by the present authors in 1998, in which our basic viewpoints on the phylogeny and evolution of angiosperms were presented. In this paper, a new outline of classification of the Magnoliophyta (angiosperms) is proposed. The Magnoliophyta are divided into 8 classes, 40 subclasses, 202 orders and 572 families. Among them, 22 new subclasses (Annonidae, Illiciidae, Ceratophyllidae, Lauridae, Calycanthidae, Chloranthidae, Aristolochiidae, Polygonidae, Plumbaginidae, Bromeliidae, Zingiberidae, Juncidae, Poaidae, Paeoniidae, Papaveridae, Trochodendridae, Betulidae, Malvidae, Ericidae, Myrtidae, Rutidae, Geraniidae) and 6 new orders (Degeneriales, Aizoales, Platanales, Dipentodontales, Meliosmales, Balanitales) are circumscribed. The number of genera and speciesin the families and each family’s geographical distribution are given.     

Vibrational distribution of nitrogen molecules in the C3Πu state in a near-surface microwave plasma in nitrogen at pressures of 1–5 Torr

The radiation of the second positive nitrogen system has been used to study the spatial dependence of the vibrational distribution of nitrogen molecules in the C³Π_u state in the near-surface plasma layer of an electrode microwave discharge in nitrogen at pressures of 1–5 Torr. It has been shown that the vibrational distribution changes at a scale of 100 μm. It has been concluded that this state is populated owing to the electron impact from the ground state. The possibility of using the local approximation for the electron energy distribution function to explain the experimental results has been analyzed.     

Prenatal diagnosis of familial hypercholesterolemia caused by the “Lebanese” mutation at the low density lipoprotein receptor locus

Summary Here, we report the prenatal diagnosis of familial hypercholesterolemia in a Christian-Arab family that carries the Lebanese mutation, a single base substitution that creates a HinfI restriction site, at the low density lipoprotein (LDL) receptor locus. Polymerase chain reaction amplification and restriction analysis were performed on genomic DNA extracted from a chorionic villus sample. In conjunction with karyotype analysis, the fetus was identified as a heterozygous female. Analysis of LDL receptor restriction fragment length polymorphisms confirmed the presence of a male parent marker and revealed that the fetus inherited the mutant gene from its mother. This technique offers a simple and rapid diagnostic tool that can be carried out at an early stage of gestation. It is recommended for families and population groups with molecularly defined LDL receptor mutations.     

Site-specific O-glycosylation of N-terminal serine residues by polypeptide GalNAc-transferase 2 modulates human δ-opioid receptor turnover at the plasma membrane

G protein-coupled receptors (GPCRs) are an important protein family of signalling receptors that govern a wide variety of physiological functions. The capacity to transmit extracellular signals and the extent of cellular response are largely determined by the amount of functional receptors at the cell surface that is subject to complex and fine-tuned regulation. Here, we demonstrate that the cell surface expression level of an inhibitory GPCR, the human δ-opioid receptor (hδOR) involved in pain and mood regulation, is modulated by site-specific N-acetylgalactosamine (GalNAc) -type O-glycosylation. Importantly, we identified one out of the 20 polypeptide GalNAc-transferase isoforms, GalNAc-T2, as the specific regulator of O-glycosylation of Ser6, Ser25 and Ser29 in the N-terminal ectodomain of the receptor. This was demonstrated by in vitro glycosylation assays using peptides corresponding to the hδOR N-terminus, Vicia villosa lectin affinity purification of receptors expressed in HEK293 SimpleCells capable of synthesizing only truncated O-glycans, GalNAc-T edited cell line model systems, and site-directed mutagenesis of the putative O-glycosylation sites. Interestingly, a single-nucleotide polymorphism, at residue 27 (F27C), was found to alter O-glycosylation of the receptor in efficiency as well as in glycosite usage. Furthermore, flow cytometry and cell surface biotinylation assays using O-glycan deficient CHO-ldlD cells revealed that the absence of O-glycans results in decreased receptor levels at the plasma membrane due to enhanced turnover. In addition, mutation of the identified O-glycosylation sites led to a decrease in the number of ligand-binding competent receptors and impaired agonist-mediated inhibition of cyclic AMP accumulation in HEK293 cells. Thus, site-specific O-glycosylation by a selected GalNAc-T isoform can increase the stability of a GPCR, in a process that modulates the constitutive turnover and steady-state levels of functional receptors at the cell surface.     

Evolution of major histocompatibility complex by “en bloc” duplication before mammalian radiation

Duplications are an important mechanism for the emergence of genetic novelties. Reports on duplicated genes are numerous, and mechanisms for polyploidization or local gene duplication are beginning to be understood. When a local duplication is studied, searches are usually done gene-by-gene, and the size of duplicated segments is not often investigated. Therefore, we do not know if the gene in question has duplicated alone or with other genes, implying that "en bloc" duplications are poorly studied. We propose a method for identification of "en bloc" duplication using mapping, phylogenetic and statistical analyses. We show that two segments present in the major histocompatibility complex (MHC) region of human chromosome 6 have resulted from an "en bloc" duplication that took place between divergence of amniotes and methaterian/eutherian separation. These segments contain members of the same multigenic families, namely olfactory receptors genes, genes encoding proteins containing B30.2 domain, genes encoding proteins containing immunoglobulin V domain and MHC class I genes. We will discuss the fact that olfactory receptors and MHC genes have undergone positive selection, which could have helped in fixation of the surrounding genes.     

Evaluation of changes in the cortical gait control in post-stroke patients induced by the use of the “Regent” soft exoskeleton complex (SEC) by navigated transcranial magnetic stimulation

The mechanisms underlying the locomotion recovery in poststroke patients remain unknown. Navigated transcranial magnetic stimulation (nTMS) is a new method to evaluate the functional state of the motor system. Using of the “Regent” soft exoskeleton complex (SEC) allow to correct walking pattern significantly. The aim of this study was to evaluate the capability of nTMS to assess changes in gait cortical control using SEC in poststroke patients. 14 patients of the subcortical stroke (the mean age was 53.0 years, the mean time from stroke onset was 14.2 months) received 10 training sessions with SEC. The patients received nTMS before and after the sessions, as well as they were clinically evaluated by the Fugl–Meyer scale section for lower extremity and a 10-m walk test. Whereas a reliable reduction in the time of walking for 10 m was recorded after the sessions with the application of SEC, the Fugl-Meyer scale assessment remained unchanged. During nTMS, a reduction was recorded in the average latency of evoked motor response from the affected hemisphere, as well as various patterns of changes in the size and localization of cortical representations of the leg muscles. We have concluded that the nTMS method allowed us to identify the individual patterns of changes in the cortical representations of leg muscles as a result of the use of SEC in post-stroke patients with injuries in a group of locomotor system elements, thus identifying not only the fact of the undergoing neuroplastic processes, but also their direction.     

The use of cycloamylose to probe the “charge-relay” system

The effect of the combination of imidazolyl and carboxyl groups on the cleavage of m-t-butylphenyl acetate in the presence of α-cyclodextrin was examined to shed light on the role of the “charge-relay” system in serine esterases. 2-Benzimidazole-acetic acid, which has both the imidazolyl and carboxyl groups in the same molecule, accelerates the cleavage of m-t-butylphenyl acetate in the presence of α-cyclodextrin. On the other hand, neither benzimidazole (which has only an imidazolyl group) nor 2-naphthaleneacetic acid (which has only a carboxyl group) exhibited measurable acceleration. The cleavage of m-t-butylphenyl acetate by the α-cyclodextrin-2-benzimidazolecetic acid system takes place through inclusion complex formation between m-t-butylphenyl acetate and α-cyclodextrin, followed by catalysis associated with the combination of the carboxyl anion, the neutral imidazolyl group, and the alkoxide anion. The most probable explanation for the combination of the three groups in the catalysis involves nucleophilic attack by the imidazolyl group, assisted by the carboxyl and alkoxide anions. The mechanism of the combination of the imidazolyl, carboxyl, and hydroxyl groups is apparently different from those shown by the “charge-relay” system in enzymatic reactions.     

Potential dependence of the “electrically silent” anion exchange across the plasma membrane ofXenopus oocytes mediated by the band-3 protein of mouse red blood cells

Summary Mouse erythroid band-3 protein was incorporated into the plasma membrane ofXenopus oocytes by microinjection of poly(A)⁺-mRNA from spleens of anemic mice. Subsequently, the efflux of microinjected³⁶Cl was continuously followed in single oocytes in a perfusion chamber the bottom of which was formed by the window of a Geiger-Müller tube. During the flux measurements, the membrane potential was clamped to different holding potentials. The efflux increased over the voltage range of –10 to –100 mV by a factor of about 1.5. Since the membrane potential cannot act as a driving force of anion exchange, it is suggested that the observed slight potential dependence is related to a recruitment of the anion-loaded transport protein by the electrical field, thereby changing the steady-state distribution between inwardly and outwardly facing anion binding sites of the transport molecules.The experimental data are discussed in terms of ping-pong kinetics, assuming that the potential dependence is primarily due to an effect of the electrical field in the membrane on the ratelimiting interconversion of inwardly and outwardly oriented anion binding sites. The results are compatible with the assumption that in the oocyte membrane the substrate-loaded band-3 molecules are preferentially inwardly oriented, and that the transition from the inwardly to the outwardly oriented conformation is associated with a reorientation of an effective charge of 0.1 elementary charge.During progesterone-induced maturation of the oocytes, several endogenous transport systems change their activity drastically. The mouse band-3 protein in the oocyte membrane also undergoes activity changes; however, these changes do not seem to involve direct regulation by specific metabolic processes. They can be explained as a consequence of the depolarization of the membrane potential associated with the maturation process.     

Imaging of spatial distributions of the millimeter wave intensity by using visible continuum radiation from a discharge in a Cs–Xe mixture. Part I: Review of the method and its fundamentals

The first part of the review is presented which is dedicated to the time-resolved method of imaging and measuring the spatial distribution of the intensity of millimeter waves by using visible continuum (VC) emitted by the positive column (PC) of a dc discharge in a mixture of cesium vapor with xenon. The review focuses on the operating principles, fundamentals, and applications of this new technique. The design of the discharge tube and experimental setup used to create a wide homogeneous plasma slab with the help of the Cs–Xe discharge at a gas pressure of 45 Torr are described. The millimeter-wave effects on the plasma slab are studied experimentally. The mechanism of microwave-induced variations in the VC brightness and the causes of violation of the local relation between the VC brightness and the intensity of millimeter waves are discussed. Experiments on the imaging of the field patterns of horn antennas and quasi-optical beams demonstrate that this technique can be used for good-quality imaging of millimeter-wave beams in the entire millimeter-wavelength band. The method has a microsecond temporal resolution and a spatial resolution of about 2 mm. Energy sensitivities of about 10 μJ/cm² in the Ka-band and about 200 μJ/cm² in the D-band have been demonstrated.