Methane excess in Arctic surface water-triggered by sea ice formation and melting.

Arctic amplification of global warming has led to increased summer sea ice retreat, which influences gas exchange between the Arctic Ocean and the atmosphere where sea ice previously acted as a physical barrier. Indeed, recently observed enhanced atmospheric methane concentrations in Arctic regions with fractional sea-ice cover point to unexpected feedbacks in cycling of methane. We report on methane excess in sea ice-influenced water masses in the interior Arctic Ocean and provide evidence that sea ice is a potential source. We show that methane release from sea ice into the ocean occurs via brine drainage during freezing and melting i.e. in winter and spring. In summer under a fractional sea ice cover, reduced turbulence restricts gas transfer, then seawater acts as buffer in which methane remains entrained. However, in autumn and winter surface convection initiates pronounced efflux of methane from the ice covered ocean to the atmosphere. Our results demonstrate that sea ice-sourced methane cycles seasonally between sea ice, sea-ice-influenced seawater and the atmosphere, while the deeper ocean remains decoupled. Freshening due to summer sea ice retreat will enhance this decoupling, which restricts the capacity of the deeper Arctic Ocean to act as a sink for this greenhouse gas.

Antarctic Sea ice--a habitat for extremophiles.

The pack ice of Earth's polar oceans appears to be frozen white desert, devoid of life. However, beneath the snow lies a unique habitat for a group of bacteria and microscopic plants and animals that are encased in an ice matrix at low temperatures and light levels, with the only liquid being pockets of concentrated brines. Survival in these conditions requires a complex suite of physiological and metabolic adaptations, but sea-ice organisms thrive in the ice, and their prolific growth ensures they play a fundamental role in polar ecosystems. Apart from their ecological importance, the bacterial and algae species found in sea ice have become the focus for novel biotechnology, as well as being considered proxies for possible life forms on ice-covered extraterrestrial bodies.

Cu(II) inhibition of the proton translocation machinery of the influenza A virus M2 protein.

The homotetrameric M2 integral membrane protein of influenza virus forms a proton-selective ion channel. An essential histidine residue (His-37) in the M2 transmembrane domain is believed to play an important role in the conduction mechanism of this channel. Also, this residue is believed to form hydrogen-bonded interactions with the ammonium group of the anti-viral compound, amantadine. A molecular model of this channel suggests that the imidazole side chains of His-37 from symmetry-related monomers of the homotetrameric pore converge to form a coordination site for transition metals. Thus, membrane currents of oocytes of Xenopus laevis expressing the M2 protein were recorded when the solution bathing the oocytes contained various transition metals. Membrane currents were strongly and reversibly inhibited by Cu2+ with biphasic reaction kinetics. The biphasic inhibition curves may be explained by a two-site model involving a fast-binding peripheral site with low specificity for divalent metal ions, as well as a high affinity site (Kdiss approximately 2 microM) that lies deep within the pore and shows rather slow-binding kinetics (kon = 18.6 +/- 0.9 M-1 s-1). The pH dependence of the interaction with the high affinity Cu2+-binding site parallels the pH dependence of inhibition by amantadine, which has previously been ascribed to protonation of His-37. The voltage dependence of the inhibition at the high affinity site indicates that the binding site lies within the transmembrane region of the pore. Furthermore, the inhibition by Cu2+ could be prevented by prior application of the reversible blocker of M2 channel activity, BL-1743, providing further support for the location of the site within the pore region of M2. Finally, substitutions of His-37 by alanine or glycine eliminated the high affinity site and resulted in membrane currents that were only partially inhibited at millimolar concentrations of Cu2+. Binding of Cu2+ to the high affinity site resulted in an approximately equal inhibition of both inward and outward currents. The wild-type protein showed very high specificity for Cu2+ and was only partially inhibited by 1 mM Ni2+, Pt2+, and Zn2+. These data are discussed in terms of the functional role of His-37 in the mechanism of proton translocation through the channel.

Exploration of the structural features defining the conduction properties of a synthetic ion channel.

The finite-difference Poisson-Boltzmann methodology was applied to a series of parallel, alpha-helical bundle models of the designed ion channel peptide Ac-(LSSLLSL)3-CONH2. This method is able to fully describe the current-voltage curves for this channel and quantitatively explains their cation selectivity and rectification. We examined a series of energy-minimized models representing different aggregation states, side-chain rotamers, and helical rotations, as well as an ensemble of structures from a molecular dynamics trajectory. Potential energies were computed for single, permeating K+ and Cl- ions at a series of positions along a central pathway through the models. A variable-electric-field Nernst-Planck electrodiffusion model was used, with two adjustable parameters representing the diffusion coefficients of K+ and Cl- to scale the individual ion current magnitudes. The ability of a given DelPhi potential profile to fit the experimental data depended strongly on the magnitude of the desolvation of the permeating ion. Below a pore radius of 3.8 A, the predicted profiles showed large energy barriers, and the experimental data could be fit only with unrealistically high values for the K+ and Cl- diffusion coefficients. For pore radii above 3.8 A, the desolvation energies were 2kT or less. The electrostatic calculations were sensitive to positioning of the Ser side chains, with the best fits associated with maximum exposure of the Ser side-chain hydroxyls to the pore. The backbone component was shown to be the major source of asymmetry in the DelPhi potential profiles. Only two of the energy-minimized structures were able to explain the experimental data, whereas an average of the dynamics structures gave excellent agreement with experimental results. Thus this method provides a promising approach to prediction of current-voltage curves from three-dimensional structures of ion channel proteins.

The role of protonation and metal chelation preferences in defining the properties of mercury-binding coiled coils.

To define the delicate interplay between metal chelation, protein folding and function in metalloproteins, a family of de novo-designed peptides was synthesized that self-assemble in aqueous solution to form two and three-stranded alpha-helical coiled coils. Each peptide contains a single Cys residue at an a or d position of the heptad repeat. Peptide association thus produces a Cys-rich coordination environment that has been used to bind Hg(II) ions. These peptides display a pH-dependent association, with trimers observed above the pKa of Glu side-chains and dimers below this value. Finite-difference Poisson-Boltzmann calculations suggest that the dimeric state decreases the unfavorable electrostatic interactions between positively charged Lys side-chains (relative to the trimer). The Cys-containing peptides bind Hg(II) in a position-dependent fashion. Cys at a positions form three-coordinate Hg complexes at high pH where the trimeric aggregation state predominates, and two-coordinate complexes at lower pH. A d position Cys, however, is only able to generate the two-coordinate complex, illustrating the difference in coordination geometry between the two positions in the coiled coil. The binding of Hg(II) was also shown to substantially increase the stability of the helical aggregates.

Two models of the influenza A M2 channel domain: verification by comparison.

BACKGROUND: The influenza M2 protein is a simple membrane protein, containing a single transmembrane helix. It is representative of a very large family of single-transmembrane helix proteins. The functional protein is a tetramer, with the four transmembrane helices forming a proton-permeable channel across the bilayer. Two independently derived models of the M2 channel domain are compared, in order to assess the success of applying molecular modelling approaches to simple membrane proteins. RESULTS: The Calpha RSMD between the two models is 1.7 A. Both models are composed of a left-handed bundle of helices, with the helices tilted roughly 15 degrees relative to the (presumed) bilayer normal. The two models have similar pore radius profiles, with a pore cavity lined by the Ser31 and Gly34 residues and a pore constriction formed by the ring of His37 residues. CONCLUSIONS: Independent studies of M2 have converged on the same structural model for the channel domain. This model is in agreement with solid state NMR data. In particular, both model and NMR data indicate that the M2 helices are tilted relative to the bilayer normal and form a left-handed bundle. Such convergence suggests that, at least for simple membrane proteins, restraints-directed modelling might yield plausible models worthy of further computational and experimental investigation.

A functionally defined model for the M2 proton channel of influenza A virus suggests a mechanism for its ion selectivity.

The M2 protein from influenza A virus forms proton-selective channels that are essential to viral function and are the target of the drug amantadine. Cys scanning was used to generate a series of mutants with successive substitutions in the transmembrane segment of the protein, and the mutants were expressed in Xenopus laevis oocytes. The effect of the mutations on reversal potential, ion currents, and amantadine resistance were measured. Fourier analysis revealed a periodicity consistent with a four-stranded coiled coil or helical bundle. A three-dimensional model of this structure suggests a possible mechanism for the proton selectivity of the M2 channel of influenza virus.

Modeling transmembrane helical oligomers.

Recently, methods for the analysis and design of water-soluble, oligomeric bundles of alpha helices, including coiled coils, have reached a high level of sophistication. These same methods may now be applied to transmembrane helical bundles. Studies of the transmembrane domains of glycophorin, phospholamban, and the M2 protein from influenza A virus exemplify this general approach.

Primary Production in Antarctic Sea Ice

A numerical model shows that in Antarctic sea ice, increased flooding in regions with thick snow cover enhances primary production in the infiltration (surface) layer. Productivity in the freeboard (sea level) layer is also determined by sea ice porosity, which varies with temperature. Spatial and temporal variation in snow thickness and the proportion of first-year ice thus determine regional differences in sea ice primary production. Model results show that of the 40 teragrams of carbon produced annually in the Antarctic ice pack, 75 percent was associated with first-year ice and nearly 50 percent was produced in the Weddell Sea.

Thermodynamic analysis of a designed three-stranded coiled coil.

The study and successful design of coiled-coil protein structural motifs have provided much insight into the rules governing protein folding and stability. In this work we use a thermodynamic approach to quantitate the rules that govern the specific oligomerization of coiled coils. We have designed a highly stable trimeric coiled coil by placing valine residues at each a position and leucine residues at each d position of the heptad repeating unit. The peptide forms a very stable trimer as determined by sedimentation equilibrium, and the concentration dependence of its circular dichroism spectrum follows a cooperative monomer/dimer/trimer equilibrium with the dimer state as a highly unstable intermediate. Its guanidinium chloride denaturation curve was collected at several peptide concentrations, and analysis of the data confirms the cooperativity of the trimerization process and provides a free energy of stabilization of - 18.4 kcal mol-1 for the trimer. The heat capacity, delta Cp, was measured by global analysis of thermal unfolding data collected at a number of guanidinium chloride concentrations. Guanidinium chloride induces cold denaturation in the thermal unfolding curves, providing a reasonably well-determined value for delta Cp of 750 cal deg-1 mol-1. This translates to a delta Cp of 8.6 cal deg-1 mol-1 per residue and corresponds well to that expected of a coiled coil with a well-defined tertiary structure.

Bacterial Standing Stock, Activity, and Carbon Production during Formation and Growth of Sea Ice in the Weddell Sea, Antarctica.

Bacterial response to formation and growth of sea ice was investigated during autumn in the northeastern Weddell Sea. Changes in standing stock, activity, and carbon production of bacteria were determined in successive stages of ice development. During initial ice formation, concentrations of bacterial cells, in the order of 1 x 10 to 3 x 10 liter, were not enhanced within the ice matrix. This suggests that physical enrichment of bacteria by ice crystals is not effective. Due to low concentrations of phytoplankton in the water column during freezing, incorporation of bacteria into newly formed ice via attachment to algal cells or aggregates was not recorded in this study. As soon as the ice had formed, the general metabolic activity of bacterial populations was strongly suppressed. Furthermore, the ratio of [H]leucine incorporation into proteins to [H]thymidine incorporation into DNA changed during ice growth. In thick pack ice, bacterial activity recovered and growth rates up to 0.6 day indicated actively dividing populations. However, biomass-specific utilization of organic compounds remained lower than in open water. Bacterial concentrations of up to 2.8 x 10 cells liter along with considerably enlarged cell volumes accumulated within thick pack ice, suggesting reduced mortality rates of bacteria within the small brine pores. In the course of ice development, bacterial carbon production increased from about 0.01 to 0.4 mug of C liter h. In thick ice, bacterial secondary production exceeded primary production of microalgae.

Stereotactic removal of foreign bodies in the maxillofacial area.

A technique is described for making it possible to localize any deep-seated radiopaque foreign body in the soft tissues and skeleton of the maxillofacial area. Using a stereotactic guide system, any point in the facial skeleton can be reached exactly, with precalculated precision, from any point outside the face. Thus, a foreign body can be exposed and subsequently removed by a surgical approach that gives minimal discomfort and carries low risk. The target probe serves the surgeon as a guide bar whilst the tissue is dissected. A precondition of successful stereotactic operations in the head and neck area is close cooperation between neurosurgeons and maxillofacial surgeons.

[Stereotactic localization and removal of foreign bodies situated deep to the facial tissues]. / Entfernung tiefliegender Fremdkörper im Gesichtsbereich nach stereotaktischer Lokalisation.

This stereotactic method permits the localization of any radiographically visible foreign body in the maxillofacial hard and soft tissues. Stereotactic needles can be inserted either via an intraoral or an extraoral approach. Use of these needles as guides during tissue preparation allows simple exposure and subsequent removal of foreign bodies. A close cooperation between neurosurgeons and maxillofacial surgeons during this procedure is recommended. The technique is explained with the aid of 2 case reports.

[Microsurgical resection of small brain tumor processes with stereotactic control]. / Mikrochirurgische Resektion kleiner Hirntumorprozesse unter stereotaktischer Führung.

From 1986 we have operated upon 13 patients with small neoplasms localised deep intracerebrally or subcortically in the central region with stereotactic guidance. Exact localisation of the tumour was one aim, whereas another one was to avoid damaging functionally important structures. In the first two patients we performed stereotactic marking of the tumour after craniotomy. The following procedure was found to be particularly useful: - determination of the tumour coordinates and planning of the approach - approach by means of a stereotactically guided probe, if necessary biopsy - stereotactically guided introduction of the leading catheter - removal of the stereotactic frame and bedding of the patient - craniotomy and microsurgical resection of the tumour This surgical procedure may be performed in one or two separate sessions. The accuracy of microsurgical tumour resection achieved by stereotactic means resulted in a remarkable reduction of postoperative neurological deficits.

[An applicator for the implantation of I125 seeds for interstitial therapy of brain tumors]. / Ein Magazin für die Implantation von J-125-Seeds zur interstitiellen Hirntumortherapie.

A compact magazine has been developed allowing a quick and easy implantation of I-125 seeds in interstitial therapy of intracranial tumors. The magazine is made of steel and has the shape of a right parallelepiped. One seed can be put each time into a movable insert. The magazine is connected to a normal application canula. It is easy to handle and contributes much to radioprotection.

Psychiatric and neuropsychological findings after stereotactic hypothalamotomy, in cases of extreme sexual aggressivity.

Report on 14 cases treated for aggressive sexual delinquency by unilateral ventromedial hypothalamotomy. Eight of them had a thorough psychiatric and psychological examination during follow-up. The following constant modifications were found: Decrease in the domination by sexual drive, increase in the fluency in semantic contexts, increase of rapidity of visual image formation and of coordinative perception processes, positive modifications in the scope of some personality dimensions (poise, openness, self-criticism), decreased colour perception, increased appetite. The structure of the patient's sexuality was not changed but the probability of a specific aggressive behaviour reduced, due to the diminished sexual drive thus allowing a positive consolidation of their social interactions with more harmonic relationships to the families or partners. These results give proof that the many prejudices and aversions against this type of therapy are not justified.

Retrogasserian glycerol injection or percutaneous stimulation in the treatment of typical and atypical trigeminal pain.

Of 164 patients with trigeminal neuralgia treated by percutaneous retrogasserian glycerol injections, we have studied the long-term outcome (one to four years) of 72 patients. 51 patients suffered from idiopathic trigeminal neuralgia, and of these 92% were freed from pain by the operation. 21 patients suffered from symptomatic trigeminal neuralgia due to multiple sclerosis or of traumatic or infectious origin. Of these, only 38% became free of pain. A further 8 patients with facial deafferentation pain who underwent treatment by retrogasserian neurostimulation with a permanently implanted electrode were included in this study. 4 of these patients were rendered free of pain by this procedure.