Physical understanding of pore formation on supported lipid bilayer by bacterial toxins

Pore forming toxins are being classified in the protein community based on their ability of forming pores in living cell membranes. Some initial study has apparently pointed out the crystallographic pathway rather can be viewed as a structural as well as morphological changes of proteins in terms of self assembly before and during the pore formation process in surfactant medium. Being a water soluble compound, it changes its conformation and originates some pre-pore complex, which later partially goes inside the cell membrane causing a pore. The physical mechanism for this whole process is still unknown. In this study we have tried to understand these types of biological processes from physical point of view by using supported lipid bilayer as a model system.

Synthesis, Characterization and Properties of Single-Walled Carbon Nanohorns

Single-walled nanohorns (SWNHs) have been prepared by sub-merged arc discharge of graphite electrodes in liquid nitrogen. The samples were examined by scanning electron microscopy, transmission electron microscopy and Raman spectroscopy. Nitrogen and boron doped SWNHs have been prepared by the sub-merged arc discharge method using melamine and elemental boron as precursors. Intensification of Raman D-band and stiffening of G-band has been observed in the doped samples. The electrical resistance of the SWNHs varies in opposite directions with nitrogen and boron doping. Functionalization of SWNHs through amidation has been carried out for solubilizing them in non-polar solvents. Water-soluble SWNHs have been produced by acid treatment and non-covalent functionalization with a coronene salt. SWNHs have been decorated with nanoparticles of Au, Ag and Pt. Interaction of electron donor (tetrathiafulvalene, TTF) and acceptor molecules (tetracyanoethylene, TCNE) with SWNHs has been investigated by Raman spectroscopy. Progressive softening and stiffening of Raman G-band has been observed respectively with increase in the concentration of TTF and TCNE.

Tripodal Bile Acid Architectures Based on a Triarylphosphine Oxide Core Obtained by Copper-Catalysed 1,3]-Dipolar Cycloaddition: Synthesis and Preliminary Aggregation Studies

We report the synthesis and aggregation behaviour of new water-soluble, bile acid derived tripodal architectures based on a core derived from triphenylphosphine oxide. We employed the well-established copper-catalysed 1,3]-dipolar cycloaddition (CuAAC) for the construction of these tripodal molecules. The aggregation behaviour of these molecules in aqueous media was studied by different analytical methods such as dye solubilisation, dynamic light scattering, NMR and AFM. These molecular architectures also offer an additional advantage in aiding understanding of the influence of the nature of the bile acid backbone and of the configuration at the steroid C-3 position in these architectures; to the best of our knowledge this has not been reported in the literature. The unique gelation properties of the -derivatives were explained through molecular modelling studies and the mechanical behaviour of these gels was studied by rheology experiments.

Green ionothermal synthesis of hierarchical nanostructures of SnS2 and their Li-ion storage properties

Flower-like hierarchical architectures of layered SnS2 have been synthesized ionothermally for the first time, using a water soluble EMIM]BF4 ionic liquid (IL) as the solvent medium. At lower reaction temperatures, the hierarchical structures are formed of few-layered polycrystalline 2D nanosheet-petals composed of randomly oriented nanoparticles of SnS2. The supramolecular networks of the IL serve as templates on which the nanoparticles of SnS2 are glued together by combined effects of hydrogen bonding, electrostatic, hydrophobic and imidazolium stacking interactions of the IL, giving rise to polycrystalline 2D nanosheet-petals. At higher reaction temperatures, single crystalline plate-like nanosheets with well-defined crystallographic facets are obtained due to rapid inter-particle diffusion across the IL. Efficient surface charge screening by the IL favors the aggregation of individual nanosheets to form hierarchical flower-like architectures of SnS2. The mechanistic aspects of the ionothermal bottom-up hierarchical assembly of SnS2 nanosheets are discussed in detail. Li-ion storage properties of the pristine SnS2 samples are examined and the electrochemical performance of the sample synthesized at higher temperatures is found to be comparable to that reported for pristine SnS2 samples in the literature.

Production and characterization of bioflocculants for mineral processing applications

A comparative study of two bacterial strains namely, Bacillus licheniformis and Bacillus firmus in the production of bioflocculants was made. The highest bioflocculant yield of 16.55 g/L was obtained from B. licheniformis (L) and 10 g/L from B. firmus (F). The bioflocculants obtained from the bacterial species were water soluble and insoluble in organic solvents. FTIR spectral analysis revealed the presence of hydroxyl, carboxyl and sugar derivatives in the bioflocculants. Thermal characterization by differential scanning calorimetry (DSC) showed the crystalline transition and the melting point (T-m) at 90-100 degrees C. Effects of bioflocculant dosage and pH on the flocculation of clay fines were evaluated. Highest bioflocculation efficiency on kaolin clay suspensions was observed at an optimum bioflocculant dosage of 5 g/L. The optimum pH range for the maximum bioflocculation was at pH 7-9. Bioflocculants exhibited high efficiency in dye decolorization. The maximum Cr (VI) removal was found to be 85 % for L (bioflocculant dosage at 2 g/L). This study demonstrates that microbial bioflocculants find potential applications in mineral processing such as selective flocculation of mineral fines, decolorization of dye solutions and in the remediation of toxic metal solutions. (C) 2015 Elsevier B.V. All rights reserved.

Binding of carboxylatopillar5]arene with alkyl and aryl ammonium salts in aqueous medium

The complex formation of alkyl ammonium salts by water-soluble carboxylatopillar5] arene (CP5A) in aqueous medium is reported. p-Xylene diammonium salt and a series of secondary alkyl ammonium salts with various alkyl groups have been prepared and investigated for complex formation. All the ammonium salts exhibit strong host-guest complexation with CP5A under neutral aqueous conditions. H-1 NMR, H-1 DOSY and 2D NOESY NMR experiments have been performed to characterize these inclusion complexes. In this study, the hydrophobic and electrostatic interactions govern the complex formation leading to the formation of pseudorotaxane species. Five pseudo2] rotaxanes and one pseudo3] rotaxane were obtained whose association constant values and stoichiometry were evaluated by an NMR titration method. The results indicate the use of ammonium salts as new complimentary synthons for CP5A in aqueous medium, adding to the repertoire of existing recognition motifs such as paraquat and 1,4-bis(pyridinium) derivatives.

Proceedings of the Workshop on the Okhotsk Sea and Adjacent Areas [Vladivostok, June 1995]

I REPORT OF THE PICES WORKSHOP ON THE OKHOTSK SEA AND ADJACENT AREAS (pdf, 0.1 Mb) 1. Outline of the workshop 2. Summary reports from sessions 3. Recommendations of the workshop 4. Acknowledgments II SCIENTIFIC PAPERS SUBMITTED FROM SESSIONS 1. Physical Oceanography Sessions (pdf, 4 Mb) A. Circulation and water mass structure of the Okhotsk Sea and Northwestern Pacific Valentina D. Budaeva & Vyacheslav G. Makarov Seasonal variability of the pycnocline in La Perouse Strait and Aniva Gulf Valentina D. Budaeva & Vyacheslav G. Makarov Modeling of the typical water circulations in the La Perouse Strait and Aniva Gulf region Nina A. Dashko, Sergey M. Varlamov, Young-Ho Han & Young-Seup Kim Anticyclogenesis over the Okhotsk Sea and its influence on weather Boris S. Dyakov, Alexander A. Nikitin & Vadim P. Pavlychev Research of water structure and dynamics in the Okhotsk Sea and adjacent Pacific Howard J. Freeland, Alexander S. Bychkov, C.S. Wong, Frank A. Whitney & Gennady I. Yurasov The Ohkotsk Sea component of Pacific Intermediate Water Emil E. Herbeck, Anatoly I. Alexanin, Igor A. Gontcharenko, Igor I. Gorin, Yury V. Naumkin & Yury G. Proshjants Some experience of the satellite environmental support of marine expeditions at the Far East Seas Alexander A. Karnaukhov The tidal influence on the Sakhalin shelf hydrology Yasuhiro Kawasaki On the formation process of the subsurface mixed water around the Central Kuril Islands Lloyd D. Keigwin Northwest Pacific paleohydrography Talgat R. Kilmatov Physical mechanisms for the North Pacific Intermediate Water formation Vladimir A. Luchin Water masses in the Okhotsk Sea Andrey V. Martynov, Elena N. Golubeva & Victor I. Kuzin Numerical experiments with finite element model of the Okhotsk Sea circulation Nikolay A. Maximenko, Anatoly I. Kharlamov & Raissa I. Gouskina Structure of Intermediate Water layer in the Northwest Pacific Nikolay A. Maximenko & Andrey Yu. Shcherbina Fine-structure of the North Pacific Intermediate Water layer Renat D. Medjitov & Boris I. Reznikov An experimental study of water transport through the Straits of Okhotsk Sea by electromagnetic method Valentina V. Moroz Oceanological zoning of the Kuril Islands area in the spring-summer period Yutaka Nagata Note on the salinity balance in the Okhotsk Sea Alexander D. Nelezin Variability of the Kuroshio Front in 1965-1991 Vladimir I. Ponomarev, Evgeny P. Varlaty & Mikhail Yu. Cheranyev An experimental study of currents in the near-Kuril region of the Pacific Ocean and in the Okhotsk Sea Stephen C. Riser, Gennady I. Yurasov & Mark J. Warner Hydrographic and tracer measurements of the water mass structure and transport in the Okhotsk Sea in early spring Konstantin A. Rogachev & Andrey V. Verkhunov Circulation and water mass structure in the southern Okhotsk Sea, as observed in summer, 1994 Lynne D. Talley North Pacific Intermediate Water formation and the role of the Okhotsk Sea Anatoly S. Vasiliev & Fedor F. Khrapchenkov Seasonal variability of integral water circulation in the Okhotsk Sea B. Sea ice and its relation to circulation and climate V.P. Gavrilo, G.A. Lebedev & A.P. Polyakov Acoustic methods in sea ice dynamics studies Nina M. Pestereva & Larisa A. Starodubtseva The role of the Far-East atmospheric circulation in the formation of the ice cover in the Okhotsk Sea Yoshihiko Sekine Anomalous Oyashio intrusion and its teleconnection with Subarctic North Pacific circulation, sea ice of the Okhotsk Sea and air temperature of the northern Asian continent C. Waves and tides Vladimir A. Luchin Characteristics of the tidal motions in the Kuril Straits George V. Shevtchenko On seasonal variability of tidal constants in the northwestern part of the Okhotsk Sea D. Physical oceanography of the Japan Sea/East Sea Mikhail A. Danchenkov, Kuh Kim, Igor A. Goncharenko & Young-Gyu Kim A “chimney” of cold salt waters near Vladivostok Christopher N.K. Mooers & Hee Sook Kang Preliminary results from a numerical circulation model of the Japan Sea Lev P. Yakunin Influence of ice production on the deep water formation in the Japan Sea 2. Fisheries and Biology Sessions (pdf, 2.8 Mb) A. Communities of the Okhotsk Sea and adjacent waters: composition, structure and dynamics Lubov A. Balkonskaya Exogenous succession of the southwestern Sakhalin algal communities Tatyana A. Belan, Yelena V. Oleynik, Alexander V. Tkalin & Tat’yana S. Lishavskaya Characteristics of pelagic and benthic communities on the North Sakhalin Island shelf Lev N. Bocharov & Vladimir K. Ozyorin Fishery and oceanographic database of Okhotsk Sea Victor V. Lapko Interannual dynamics of the epipelagic ichthyocen structure in the Okhotsk Sea Valentina I. Lapshina Quantitative seasonal and year-to-year changes of phytoplankton in the Okhotsk Sea and off Kuril area of the Pacific Lyudmila N. Luchsheva Biological productivity in anomalous mercury conditions (northern part of Okhotsk Sea) Inna A. Nemirovskaya Origin of hydrocarbons in the ecosystems of coastal region of the Okhotsk Sea Tatyana A. Shatilina Elements of the Pacific South Kuril area ecosystem Vyacheslav P. Shuntov & Yelena P. Dulepova Biota of the Okhotsk Sea: Structure of communities, the interannual dynamics and current status B. Abundance, distribution, dynamics of the common fishes of the Okhotsk Sea Yuri P. Diakov Influence of some abiotic factors on spatial population dynamics of the West Kamchatka flounders (Pleuronectidae) Gordon A. McFarlane, Richard J. Beamish & Larisa M. Zverkova An examination of age estimates of walleye pollock (Theragra chalcogramma) from the Sea of Okhotsk using the burnt otolith method and implications for stock assessment and management Larisa P. Nikolenko Migration of Greenland turbot (Reinhardtius hippoglossoides) in the Okhotsk Sea Galina M. Pushnikova Fisheries impact on the Sakhalin-Hokkaido herring population Vidar G. Wespestad Is pollock overfished? C. Salmon of the Okhotsk Sea: biology, abundance and stock identification Vladimir A. Belyaev, Alexander Yu. Zhigalin Epipelagic Far Eastern sardine of the Okhotsk Sea Yuri E. Bregman, Victor V. Pushnikov, Lyudmila G. Sedova & Vladimir Ph. Ivanov A preliminary report on stock status and productive capacity of horsehair crab Erimacrus isenbeckii (Brandt) in the South Kuril Strait Natalia T. Dolganova Mezoplankton distribution in the West Japan Sea Vladimir V. Efremov, Richard L. Wilmot, Christine M. Kondzela, Natalia V. Varnavskaya, Sharon L. Hawkins & Maria E. Malinina Application of pink and chum salmon genetic baseline to fishery management Vyacheslav N. Ivankov & Valentina V. Andreyeva Strategy for culture, breeding and numerous dynamics of Sakhalin salmon populations Alla M. Kovalevskaya, Natalia I. Savelyeva & Dmitry M. Polyakov Primary production in Sakhalin shelf waters Tatyana N. Krupnova Some reasons for resource reduction of Laminaria japonica (Primorye region) Lyudmila N. Luchsheva & Anatoliy I. Botsul Mercury in bottom sediments of the northeastern Okhotsk Sea Pavel A. Luk’yanov, Natalia I. Belogortseva, Alexander A. Bulgakov, Alexander A. Kurika & Olga D. Novikova Lectins and glycosidases from marine macro and micro-organisms of Japan and Okhotsk Seas Boris A. Malyarchuk, Olga A. Radchenko, Miroslava V. Derenko, Andrey G. Lapinski & Leonid L. Solovenchuk PCR-fingerprinting of mitochondrial genome of chum salmon, Oncorhynchus keta Alexander A. Mikheev Chaos and relaxation in dynamics of the pink salmon (Oncorhynchus gorbuscha) returns for two regions Yuri A. Mitrofanov & Larisa N. Lesnikova Fish-culture of Pacific Salmons increases the number of heredity defects Larisa P. Nikolenko Abundance of young halibut along the West Kamchatka shelf in 1982-1992 Sergey A. Nizyaev Living conditions of golden king crab Lithodes aequispina in the Okhotsk Sea and near the Kuril Islands Ludmila A. Pozdnyakova & Alla V. Silina Settlements of Japanese scallop in Reid Pallada Bay (Sea of Japan) Galina M. Pushnikova Features of the Southwest Okhotsk Sea herring Vladimir I. Radchenko & Igor I. Glebov Present state of the Okhotsk herring stock and fisheries outlook Alla V. Silina & Ida I. Ovsyannikova Distribution of the barnacle Balanus rostratus eurostratus near the coasts of Primorye (Sea of Japan) Galina I. Victorovskaya Dependence of urchin Strongylocentrotus intermedius reproduction on water temperature Anatoly F. Volkov, Alexander Y. Efimkin & Valery I. Chuchukalo Feeding habits of Pacific salmon in the Sea of Okhotsk and in the Pacific waters of Kuril Islands in summer 1993 Larisa M. Zverkova & Georgy A. Oktyabrsky Okhotsk Sea walleye pollock stock status Tatyana N. Zvyagintseva, Elena V. Sundukova, Natalia M. Shevchenko & Ludmila A. Elyakova Water soluble polysaccharides of some Far-Eastern seaweeds 3. Biodiversity Program (pdf, 0.2 Mb) A. Biodiversity of island ecosystems and seasides of the North Pacific Larissa A. Gayko Productivity of Japanese scallop Patinopecten yessoensis (IAY) culture in Posieta Bay (Sea of Japan) III APPENDICES 1. List of acronyms 2. List of participants (Document pdf contains 431 pages)

A Comparison of Two Methods for Enhancing the Recovery of Seagrasses into Propellor Scars: Mechanical Injection of a Nutrient and Growth Hormone Solution vs. Defecation by Roosting Seabirds: Final Report.

Based on the recovery rates for Thalassia testudinum measured in this study for scars of these excavation depths and assuming a linear recovery horizon, we estimate that it would take ~ 6.9 years (95% CI. = 5.4 to 9.6 years) for T. testudinum to return to the same density as recorded for the adjacent undisturbed population. The application of water soluble fertilizers and plant growth hormones by mechanical injection into the sediments adjacent to ten propellor scars at Lignumvitae State Botanical Site did not significantly increase the recovery rate of Thalassia testudinum or Halodule wrightii. An alternative method of fertilization and restoration of propellor scars was also tested by a using a method of “compressed succession” where Halodule wrightii is substituted for T. testudinum in the initial stages of restoration. Bird roosting stakes were placed among H.wrightii bare root plantings in prop scars to facilitate the defecation of nitrogen and phosphorus enriched feces. In contrast to the fertilizer injection method, the bird stakes produced extremely high recovery rates of transplanted H. wrightii. We conclude that use of a fertilizer/hormone injection machine in the manner described here is not a feasible means of enhancing T. testudinum recovery in propellor scars on soft bottom carbonate sediments. Existing techniques such as the bird stake approach provide a reliable, and inexpensive alternative method that should be considered for application to restoration of seagrasses in these environments. Document contains 40 pages)

Mechanism of transport of saquinavir-loaded nanostructured lipid carriers across the intestinal barrier

[EN] The aims of this work were (i) to evaluate the potential of nanostructured lipid carriers (NLCs) as a tool to 24 enhance the oral bioavailability of poorly soluble compounds using saquinavir (SQV), a BCS class IV drug 25 and P-gp substrate as a model drug, and (ii) to study NLC transport mechanisms across the intestinal barrier. 26 Three different NLC formulations were evaluated. SQV transport across Caco-2 monolayers was enhanced up 27 to 3.5-fold by NLCs compared to SQV suspension. M cells did not enhance the transport of NLCs loaded with 28 SQV. The size and amount of surfactant in the NLCs influenced SQV's permeability, the transcytosis pathway 29 and the efflux of SQV by P-gp. An NLC of size 247 nm and 1.5% (w/v) surfactant content circumvented P-gp 30 efflux and used both caveolae- and clathrin-mediated transcytosis, in contrast to the other NLC formulations, 31 which used only caveolae-mediated transcytosis. By modifying critical physicochemical parameters of the 32 NLC formulation, we were thus able to overcome the P-gp drug efflux and alter the transcytosis mechanism 33 of the nanoparticles. These findings support the use of NLCs approaches for oral delivery of poorly 34 water-soluble P-gp substrates.

Nachweis von Parvalbuminen in Störgeweben

Parvalbumins, calcium-binding, heat-stable proteins of considerable allergenic potential, are occurring in high concentration in light muscle of many fish species. Lack of knowledge about parvalbumins in sturgeon tissues prompted us to study the distribution of this type of proteins in light and dark muscle, as well as in swim bladder and skin of Atlantic sturgeon, Acipenser oxyrhynchus, and three other sturgeon species. Results: Light and dark muscle of sturgeons contained water soluble proteins with following properties: (1) Great stability against heating at 70 or 80 °C. (2) Isolelectric points between 3.85 and 5.66. (3) Treatment of proteins separated by isoelectric focusing with the dye “stains all” gave blue protein bands. (4) The molecular mass was about 10 to 14 kDalton. (5) Concentration of acidic, heat-stable proteins was higher in light than in dark muscle. Taken together, these findings gave strong indication for the presence of parvalbumins in muscle tissue of sturgeons. This conclusion was corroborated by immunological tests. However, parvalbumin could not be detected in swim bladder tissue. Skin preparations showed only traces of parvalbumins, possibly resulting from residual muscle tissue.

Design and synthesis of biofunctional magnetic/fluorescent glyco-nanoparticles and quantum dots and their application as specific molecular imaging probes

206 p.

The design and synthesis of a true, heterogeneous, asymmetric catalyst

This work describes the design and synthesis of a true, heterogeneous, asymmetric catalyst. The catalyst consists of a thin film that resides on a high-surface- area hydrophilic solid and is composed of a chiral, hydrophilic organometallic complex dissolved in ethylene glycol. Reactions of prochiral organic reactants take place predominantly at the ethylene glycol-bulk organic interface.

The synthesis of this new heterogeneous catalyst is accomplished in a series of designed steps. A novel, water-soluble, tetrasulfonated 2,2'-bis (diphenylphosphino)-1,1'-binaphthyl (BINAP-4S0_3Na) is synthesized by direct sulfonation of 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP). The rhodium (I) complex of BINAP-4SO_3Na is prepared and is shown to be the first homogeneous catalyst to perform asymmetric reductions of prochiral 2-acetamidoacrylic acids in neat water with enantioselectivities as high as those obtained in non-aqueous solvents. The ruthenium (II) complex, [Ru(BINAP-4SO_3Na)(benzene)Cl]Cl is also synthesized and exhibits a broader substrate specificity as well as higher enantioselectivities for the homogeneous asymmetric reduction of prochiral 2-acylamino acid precursors in water. Aquation of the ruthenium-chloro bond in water is found to be detrimental to the enantioselectivity with some substrates. Replacement of water by ethylene glycol results in the same high e.e's as those found in neat methanol. The ruthenium complex is impregnated onto a controlled pore-size glass CPG-240 by the incipient wetness technique. Anhydrous ethylene glycol is used as the immobilizing agent in this heterogeneous catalyst, and a non-polar 1:1 mixture of chloroform and cyclohexane is employed as the organic phase.

Asymmetric reduction of 2-(6'-methoxy-2'-naphthyl)acrylic acid to the non-steroidal anti-inflammatory agent, naproxen, is accomplished with this heterogeneous catalyst at a third of the rate observed in homogeneous solution with an e.e. of 96% at a reaction temperature of 3°C and 1,400 psig of hydrogen. No leaching of the ruthenium complex into the bulk organic phase is found at a detection limit of 32 ppb. Recycling of the catalyst is possible without any loss in enantioselectivity. Long-term stability of this new heterogeneous catalyst is proven by a self-assembly test. That is, under the reaction conditions, the individual components of the present catalytic system self-assemble into the supported-catalyst configuration.

The strategies outlined here for the design and synthesis of this new heterogeneous catalyst are general, and can hopefully be applied to the development of other heterogeneous, asymmetric catalysts.

New materials for biological applications prepared by olefin metathesis reactions

With the advent of well-defined ruthenium olefin metathesis catalysts that are highly active and stable to a variety of functional groups, the synthesis of complex organic molecules and polymers is now possible; this is reviewed in Chapter 1. The majority of the rest of this thesis describes the application of these catalysts towards the synthesis of novel polymers that may be useful in biological applications and investigations into their efficacy.

A method was developed to produce polyethers by metathesis, and this is described in Chapters 2 and 3. An unsaturated 12-crown-4 analog was made by template- directed ring-closing metathesis (RCM) and utilized as a monomer for the synthesis of unsaturated polyethers by ring-opening metathesis polymerization (ROMP). The yields were high and a range of molecular weights was accessible. In a similar manner, substituted polyethers with various backbones were synthesized: polymers with benzo groups along the backbone and various concentrations of amino acids were prepared. The results from in vitro toxicity tests of the unsubstituted polyethers are considered.

The conditions necessary to synthesize polynorbornenes with pendent bioactive peptides were explored as illustrated in Chapter 4. First, the polymerization of various norbornenyl monomers substituted with glycine, alanine or penta(ethylene glycol) is described. Then, the syntheses of polymers substituted with peptides GRGD and SRN, components of a cell binding domain of fibronectin, using newly developed ruthenium initiators are discussed.

In Chapter 5, the syntheses of homopolymers and a copolymer containing GRGDS and PHSRN, the more active forms of the peptides, are described. The ability of the polymers to inhibit human dermal fibroblast cell adhesion to fibronectin was assayed using an in vitro competitive inhibition assay, and the results are discussed. It was discovered that the copoymer substituted with both GRGDS and PHSR peptides was more active than both the GRGDS-containing homopolymer and the GRGDS free peptide.

Historically, one of the drawbacks to using metathesis is the removal of the residual ruthenium at the completion of the reaction. Chapter 6 describes a method where the water soluble tris(hydroxymethyl)phosphine is utilized to facilitate the removal of residual ruthenium from RCM reaction products.

Tecnologia mais limpa aplicada ao tratamento de água em sistemas de resfriamento abertos com recirculação

Os estudos destinados à aplicação de uma tecnologia mais limpa ao tratamento de águas em sistemas de resfriamento abertos com recirculação resultaram no desenvolvimento de uma filosofia de tratamento baseada na busca de uma operação com ciclo de concentração ideal. Assim, por meio do controle do balanço material no sistema de resfriamento, seria alcançado um nível de concentração em que a água de circulação apresentaria, naturalmente e ao mesmo tempo, baixa tendência à corrosão do ferro e baixa tendência à formação de incrustação de carbonato de cálcio. O tratamento com inibidores de corrosão e de formação de depósitos seria apenas complementar e, conseqüentemente, reduzido o emprego de produtos químicos para o condicionamento da água. O presente trabalho procura demonstrar que o critério de tratamento baseado apenas no índice de estabilidade não leva em conta o fato de que as espécies solúveis em água não se concentram de uma mesma forma, principalmente como decorrência de reações de conversão envolvendo o íon bicarbonato e de precipitação do íon cálcio. Como suporte desse trabalho, foram realizados ensaios de campo em um sistema de resfriamento aberto com recirculação em operação normal, porém sem tratamento químico da água. Foi possível, desse modo, observar a variação da concentração de cada espécie crítica presente na água com a vazão de descarga efetuada. Os resultados obtidos e o estudo realizado mostram que a metodologia tradicional pode apresentar desvios da realidade que podem comprometer a qualidade do tratamento, obrigando o emprego de maiores concentrações de produtos químicos para condicionamento da água, contrariando, assim, o próprio propósito original de execução de um tratamento dentro dos princípios de uma produção mais limpa. Este trabalho procura, ainda, salientar a importância qualitativa e quantitativa das espécies presentes, na água de reposição, na qualidade do tratamento, alertando que o emprego de reúso de água, como reposição de um sistema de resfriamento, deve ser criteriosamente analisado, pois uma água de má qualidade pode tornar o tratamento interno impraticável, exigindo a execução de um tratamento prévio muitas vezes inviável do ponto de vista econômico e causador de impacto ambiental

The isolation and partial biochemical analysis of Sindbis virus proteins

Recently, the amino acid sequences have been reported for several proteins, including the envelope glycoproteins of Sindbis virus, which all probably span the plasma membrane with a common topology: a large N-terminal, extracellular portion, a short region buried in the bilayer, and a short C-terminal intracellular segment. The regions of these proteins buried in the bilayer correspond to portions of the protein sequences which contain a stretch of hydrophobic amino acids and which have other common characteristics, as discussed. Reasons are also described for uncertainty, in some proteins more than others, as to the precise location of some parts of the sequence relative to the membrane.

The signal hypothesis for the transmembrane translocation of proteins is briefly described and its general applicability is reviewed. There are many proteins whose translocation is accurately described by this hypothesis, but some proteins are translocated in a different manner.

The transmembraneous glycoproteins E1 and E2 of Sindbis virus, as well as the only other virion protein, the capsid protein, were purified in amounts sufficient for biochemical analysis using sensitive techniques. The amino acid composition of each protein was determined, and extensive N-terminal sequences were obtained for E1 and E2. By these techniques E1 and E2 are indistinguishable from most water soluble proteins, as they do not contain an obvious excess of hydrophobic amino acids in their N-terminal regions or in the intact molecule.

The capsid protein was found to be blocked, and so its N-terminus could not be sequenced by the usual methods. However, with the use of a special labeling technique, it was possible to incorporate tritiated acetate into the N-terminus of the protein with good specificity, which was useful in the purification of peptides from which the first amino acids in the N-terminal sequence could be identified.

Nanomole amounts of PE2, the intracellular precursor of E2, were purified by an immuno-affinity technique, and its N-terminus was analyzed. Together with other work, these results showed that PE2 is not synthesized with an N-terminal extension, and the signal sequence for translocation is probably the N-terminal amino acid sequence of the protein. This N-terminus was found to be 80-90% blocked, also by Nacetylation, and this acetylation did not affect its function as a signal sequence. The putative signal sequence was also found to contain a glycosylated asparagine residue, but the inhibition of this glycosylation did not lead to the cleavage of the sequence.