Disruption of Saccharomyces cerevisiae by Plantaricin 149 and investigation of its mechanism of action with biomembrane model systems

The action of a synthetic antimicrobial peptide analog of Plantaricin 149 (Pln149a) against Saccharomyces cerevisiae and its interaction with biomembrane model systems were investigated. Pln149a was shown to inhibit S. cerevisiae growth by more than 80% in YPD medium, causing morphological changes in the yeast wall and remaining active and resistant to the yeast proteases even after 24 h of incubation. Different membrane model systems and carbohydrates were employed to better describe the Pln149a interaction with cellular components using circular dichroism and fluorescence spectroscopies, adsorption kinetics and surface elasticity in Langmuir monolayers. These assays showed that Pln149a does not interact with either mono/polysaccharides or zwitterionic LUVs, but is strongly adsorbed to and incorporated into negatively charged surfaces, causing a conformational change in its secondary structure from random-coil to helix upon adsorption. From the concurrent analysis of Pln149a adsorption kinetics and dilatational surface elasticity data, we determined that 2.5 mu M is the critical concentration at which Pln149a will disrupt a negative DPPG monolayer. Furthermore, Pln149a exhibited a carpet-like mechanism of action, in which the peptide initially binds to the membrane, covering its surface and acquiring a helical structure that remains associated to the negatively charged phospholipids. After this electrostatic interaction, another peptide region causes a strain in the membrane, promoting its disruption. (C) 2009 Elsevier B.V. All rights reserved.

Four novel mononuclear and polynuclear Cu(I) thiosaccharinates with triphenylphosphane and bis(diphenylposphino)methane. Synthesis and structural study of Cu(4)(tsac)(4)(PPh(3))(3), Cu(tsac)(PPh(3))(2), Cu(4)(tsac)(4)(dppm)(2), and Cu(2)(tsac)(2)(dppm)(2)

Four new ternary complexes of copper(I) with thiosaccharin and phosphanes were prepared. The reaction of [Cu(4)(tsac)(4)(CH(3)CN)(2)] (1) (tsac: thiosaccharinate anion) with PPh(3) in molar ratios Cu(I)/PPh(3) 1:075 and 1:2 gave the complexes [Cu(4)(tsac)(4)(PPh(3))(3)] center dot CH(3)CN (2) and Cu(tsac)(PPh(3))(2) (3), respectively. The reaction of 1 with Ph(2)PCH(2)PPh(2) (dppm) in molar ratios Cu(I)/dppm 2:1 and 1:1 gave the complexes [Cu(4) (tsac)(4)(dppm)(2)] center dot 2CH(2)Cl(2) (4) and [Cu(2)(tsac)(2)(dppm)(2)] center dot CH(2)Cl(2) (5), respectively. All the compounds have been characterized by spectroscopic and X-ray crystallographic methods. Complex 2 presents a tetra-nuclear arrangement with three metal centers in distorted tetrahedral S(2)NP environments, the fourth one with the Cu(I) ion in a distorted trigonal S(2)N coordination sphere, and the tsac anions acting as six electron donor ligands in mu(3)-S(2)N coordination forms. Complex 3 shows mononuclear molecular units with copper(I) in a distorted trigonal planar coordination sphere, built with the exocyclic S atom of a mono-coordinated thiosaccharinate anion and two P-atoms of triphenylphosphane molecules. With dppm as secondary ligand the structures of the complexes depends strongly on the stoicheometry of the preparation reaction. Complex 4 has a centrosymmetric structure. Two triply bridged Cu(2)(tsac)(2)(dppm) units are joined together by the exocyclic S-atoms of two tsac anions acting effectively as bridging tridentate ligands. Complex 5 is conformed by asymmetric dinuclear moieties where the two dppm and one tsac ligands bridge two Cu(I) atoms and the second tsac anion binds one of the metal centers through its exocyclic S-atom. (C) 2009 Elsevier B.V. All rights reserved.

Interaction of cationic bilayer fragments with a model oligonucleotide

The interaction between cationic bilayer fragments and a model oligonucleotide was investigated by differential scanning calorimetry, turbidimetry, determination of excimer to monomer ratio of 2-(10-(1-pyrene)-decanoyl)-phosphatidyl-choline in bilayer fragment dispersions and dynamic light scattering for sizing and zeta-potential analysis. Salt (Na(2)HPO(4)), mononucleotide (2`-deoxyadenosine-5`-monophosphate) or poly (dA) oligonucleotide (3`-AAA AAA AAA A-5`) affected structure and stability of dioctadecyldimethylammonium bromide bilayer fragments. Oligonucleotide and salt increased bilayer packing due to bilayer fragment fusion. Mononucleotide did not reduce colloid stability or did not cause bilayer fragment fusion. Charge neutralization of bilayer fragments by poly (dA) at 1:10 poly (dA):dioctadecyldimethylammonium bromide molar ratio caused extensive aggregation, maximal size and zero of zeta-potential for the assemblies. Above charge neutralization, assemblies recovered colloid stability due to charge overcompensation. For bilayer fragments/poly (dA), the nonmonotonic behavior of colloid stability as a function of poly (dA) concentration was unique for the oligonucleotide and was not observed for Na(2)HPO(4) or 2`-deoxyadenosine-5`-monophosphate. For the first time, such interactions between cationic bilayer fragments and mono- or oligonucleotide were described in the literature. Bilayer fragments/oligonucleotide assemblies may find interesting applications in drug delivery. (c) 2010 Elsevier B.V. All rights reserved.

Purification and partial characterization of a midgut membrane-bound alpha-glucosidase from Quesada gigas (Hemiptera: Cicadidae)

Adults of Quesada gigas (Hemiptera: Cicadidae) have a major alpha-glucosidase bound to the perimicrovillar membranes, which are lipoprotein membranes that surround the midgut cell microvilli in Hemiptera and Thysanoptera. Determination of the spatial distribution of alpha-glucosidases in Q. gigas midgut showed that this activity is not equally distributed between soluble and membrane-bound isoforms. The major membrane-bound enzyme was solubilized in the detergent Triton X-100 and purified to homogeneity by means of gel filtration on Sephacryl S-100, and ion-exchange on High Q and Mono Q columns. The purified alpha-glucosidase is a protein with a pH optimum of 6.0 against the synthetic substrate p-nitrophenyl alpha-D-glucoside and M(r) of 61,000 (SDS-PAGE). Taking into account V(Max)/K(M) ratios, the enzyme is more active on maltose than sucrose and prefers oligomaltodextrins up to maltopentaose, with lower efficiency for longer chain maltodextrins. The Q gigas alpha-glucosidase was immunolocalized in perimicrovillar membranes by using a monospecific polyclonal antibody raised against the purified enzyme from Dysdercus peruvianus. The role of this enzyme in xylem fluid digestion and its possible involvement in osmoregulation is discussed. (C) 2009 Elsevier Inc. All rights reserved.

PREDICTION OF METAL CATION TOXICITY TO THE BIOLUMINESCENT FUNGUS GERRONEMA VIRIDILUCENS

A correlation between the physicochemical properties of mono- [Li(I), K(I), Na(I)] and divalent [Cd(II), Cu(II), Mn(II), Ni(II), Co(II), Zn(II), Mg(II), Ca(II)] metal cations and their toxicity (evaluated by the free ion median effective concentration. EC50(F)) to the naturally bioluminescent fungus Gerronema viridilucens has been studied using the quantitative ion character activity relationship (QICAR) approach. Among the 11 ionic parameters used in the current study, a univariate model based on the covalent index (X(m)(2)r) proved to be the most adequate for prediction of fungal metal toxicity evaluated by the logarithm of free ion median effective concentration (log EC50(F)): log EC50(F) = 4.243 (+/-0.243) -1.268 (+/-0.125).X(m)(2)r (adj-R(2) = 0.9113, Alkaike information criterion [AIC] = 60.42). Additional two- and three-variable models were also tested and proved less suitable to fit the experimental data. These results indicate that covalent bonding is a good indicator of metal inherent toxicity to bioluminescent fungi. Furthermore, the toxicity of additional metal ions [Ag(I), Cs(I), Sr(II), Ba(II), Fe(II), Hg(II), and Pb(II)] to G. viridilucens was predicted, and Pb was found to be the most toxic metal to this bioluminescent fungus (EC50(F)): Pb(II) > Ag(I) > Hg(I) > Cd(II) > Cu(II) > Co(II) Ni(II) > Mn(II) > Fe(II) approximate to Zn(II) > Mg(II) approximate to Ba(II) approximate to Cs(I) > Li(I) > K(I) approximate to Na(I) approximate to Sr(II)> Ca(II). Environ. Toxicol. Chem. 2010;29:2177-2181. (C) 2010 SETAC

Photophysics and photostability of adenine in aqueous solution: A theoretical study

The sequential Monte Carlo/CASPT2 approach was employed to investigate deactivation and emission processes from the lowest-lying pi pi * and n pi * excited states of 9H-adenine in aqueous solution. It is found that conical intersections connecting the pi pi* and n pi* states with the ground state are also present in solution, whereas the barriers for the deactivation paths are significantly smaller on solvated conditions. The large destabilization of the n pi* state found in solution possibly prevents its involvement in the deactivation photophysics and explains the change from a bi- to a mono-exponential decay for the molecule in the gas phase and solution, respectively. (C) 2010 Elsevier B.V. All rights reserved.

Metabolism of (-)-grandisin from Piper solmsianum in Coleoptera and Lepidoptera species

The biotransformation of the major Piper solmsianum leaf phenylpropanoids, such as the tetrahydrofuran lignan grandisin and derivatives was investigated in the beetle Naupactus bipes as well as in the caterpillars Heraclides hectorides and Quadrus u-lucida. Analysis of fecal material indicated that metabolism occurred mainly through mono- and di-O-demethylation at para positions of 3,4,5-trimethoxyphenyl rings of tetrahydrofuran lignans during digestion by these insects. Additionally, 3-hydroxy-4,5-dimethoxycinnamyl and 3,4,5-trimethoxycinnamyl alcohols were identified in fecal extracts of N. bipes. (C) 2008 Elsevier Ltd. All rights reserved.

Solvatochromism in Binary Mixtures: First Report on a Solvation Free Energy Relationship between Solvent Exchange Equilibrium Constants and the Properties of the Medium

We have employed UV-vis spectroscopy in order to investigate details of the solvation of six solvatochromic indicators, hereafter designated as ""probes"", namely, 2,6-diphenyl-4-(2,4,6-triphenylpyridinium-1-yl) phenolate (RB); 4-[(E)-2-(1-methylpyridinium-4-yl)ethenyl] phenolate, MePM; 1-methylquinolinium-8-olate, QB; 2-bromo-4-[(E)-2-(1-methylpyridinium-4-yl)ethenyl] phenolate, MePMBr, 2,6-dichloro-4-(2,4,6-triphenylpyridinium-1-yl) phenolate (WB); and 2,6-dibromo-4-[(E)-2-(1-methylpyridinium-4-yl)ethenyl] phenolate, MePMBr,, respectively. These can be divided into three pairs, each includes two probes of similar pK(a) in water and different lipophilicity. Solvation has been studied in binary mixtures, BMs, of water, W, with 12 protic organic solvents, S, including mono- and bifunctional alcohols (2-alkoxyethanoles, unsaturated and chlorinated alcohols). Each medium was treated as a mixture of S, W, and a complex solvent, S-W, formed by hydrogen bonding. Values of lambda(max) (of the probe intramolecular charge transfer) were converted into empirical polarity scales, E(T)(probe) in kcal/mol, whose values were correlated with the effective mole fraction of water in the medium, chi w(effective). This correlation furnished three equilibrium constants for the exchange of solvents in the probe solvation shell; phi(W/S) (W substitutes S): phi(S-W/W) (S-W substitutes W), and phi(S-W/S) (S-W substitutes S), respectively. The values of these constants depend on the physicochemical properties of the probe and the medium. We tested, for the first time, the applicability of a new solvation free energy relationship: phi = constant + a alpha(BM) + b beta(BM) + s(pi*(BM) + d delta) + p log P(BM), where a, b, s, and p are regression coefficients alpha(BM), beta(BM), and pi*(BM) are solvatochromic parameters of the BM, delta is a correction term for pi*, and log P is an empirical scale of lipophilicity. Correlations were carried out with two-, three-, and four-medium descriptors. In all cases, three descriptors gave satisfactory correlations; use of four parameters gave only a marginal increase of the goodness of fit. For phi(W/S), the most important descriptor was found to be the lipophilicity of the medium; for phi(S-W/W) and phi(S-W/S), solvent basicity is either statistically relevant or is the most important descriptor. These responses are different from those of E(T)(probe) of many solvatochromic indicators in pure solvents, where the importance of solvent basicity is usually marginal, and can be neglected.

Self-reinforced composites obtained by the partial oxypropylation of cellulose fibers. 1. Characterization of the materials obtained with different types of fibers

The in-depth oxypropylation of different types of cellulose fibers, namely Avicel, Rayon, Kraft, and Filter Paper, was investigated. New biphasic mono-component materials were obtained, which could be hot-pressed to form films of cellulose fibers dispersed into a thermoplastic matrix. The success of this chemical modification was assessed by FTIR spectroscopy, X-ray diffraction, scanning electron microscopy. differential scanning calorimetry, thermogravimetric analysis and contact angle measurements. The optimization of this process led to the establishment of the optimal molar ratio between the cellulose CH groups and propylene oxide, which varied as a function of the specific morphology of the fibers. (C) 2008 Elsevier Ltd. All rights reserved.

Förutsättningar för specialiserade musikproduktioner för personer med cochleaimplantat

Denna studies syfte är att finna sätt att producera musikinspelningar som passar lyssnare med cochleaimplantat. Frågeställningen är: Vilka perceptuella egenskaper hos cochleaimplantat bör man som musikproducent tänka på för att kunna anpassa musikinspelningar för människor med CI?För att finna de perceptuella egenskaper som cochleaimplantat, CI, har vid musiklyssning gjordes en litteraturstudie av det som skrivits kopplat till musiklyssning med CI.Resultaten visar att inspelningarna bör mixas i mono och att melodierna i musiken bör innehålla toner med stora intervall. Grundtonen i komplexa toner bör höras, och övertonerna bör kontrolleras, så att övertonsserierna inte blir alltför långa och går för högt i frekvensspektrumet. Det dynamiska omfånget i inspelningen bör inte vara alltför stort, och frekvensomfånget bör hålla sig runt 200-5000 Hz. Syntetisk rumssimulering, som t.ex. reverb, bör inte heller användas. Vidare presenteras en idé till hur man skulle kunna styra implantaten direkt elektroniskt, för att skapa musik.

Photovoltaic Process Development and innovative Techniques

Photovoltaic processing is one of the processes that have significance in semiconductor process line. It is complicated due to the no. of elements involved that directly or indirectly affect the processing and final yield. So mathematically or empirically we can’t say assertively about the results specially related with diffusion, antireflective coating and impurity poisoning. Here I have experimented and collected data on the mono-crystal silicon wafers with varying properties and outputs. Then by using neural network with available experimental data output required can be estimated which is further tested by the test data for authenticity. One can say that it’s a kind of process simulation with varying input of raw wafers to get desired yield of photovoltaic mono-crystal cells.

Buried screen-printed contacts for silicon solar cells

A Simple way to improve solar cell efficiency is to enhance the absorption of light and reduce the shading losses. One of the main objectives for the photovoltaic roadmap is the reduction of metalized area on the front side of solar cell by fin lines. Industrial solar cell production uses screen-printing of metal pastes with a limit in line width of 70-80 μm. This paper will show a combination of the technique of laser grooved buried contact (LGBC) and Screen-printing is able to improve in fine lines and higher aspect ratio. Laser grooving is a technique to bury the contact into the surface of silicon wafer. Metallization is normally done with electroless or electrolytic plating method, which a high cost. To decrease the relative cost, more complex manufacturing process was needed, therefore in this project the standard process of buried contact solar cells has been optimized in order to gain a laser grooved buried contact solar cell concept with less processing steps. The laser scribing process is set at the first step on raw mono-crystalline silicon wafer. And then the texturing etch; phosphorus diffusion and SiNx passivation process was needed once. While simultaneously optimizing the laser scribing process did to get better results on screen-printing process with fewer difficulties to fill the laser groove. This project has been done to make the whole production of buried contact solar cell with fewer steps and could present a cost effective opportunity to solar cell industries.

O mutante pso9-1 de Saccharomyces cerevisiae, sensível a psoralenos fotoativados, contém um alelo mutante do gene MEC3 envolvido em checkpoint

Análises de complementação do mutante pso9-1, sensível a psoralenos mono- e bifuncionais fotoativados, UV254nm e nitrosoguanidina, com os mutantes pso1 a pso8, confirmou que este contém uma nova mutação pso. A clonagem molecular a partir de um banco genômico de levedura sugeriu pso9-1 como alelo mutante do gene de checkpoint que responde a danos no DNA MEC3. A não-complementação de vários fenótipos de sensibilidade em diplóides pso9-1/mec3∆ confirmou o alelismo dos dois mutantes. A sensibilidade à calcofluor white e à cafeína não foi aumentada nas linhagens pso9-1 e mec3∆, em relação as suas respectivas selvagens, sugerindo que o mutante pso9-1 não porta uma mutação na região promotora. O fenótipo mutante de mec3∆ foi levemente mais pronunciado para sensibilidade à 8-MOP + UVA e UVC, e para a supressão de mutação para frente induzida por UVC, sugerindo uma função residual para a proteína produzida por este alelo mutante.

Caracterização molecular da mutação pso8-1/rad6 de Saccharomyces cerevisiae, sensível à fotoadição de psoralenos, à radiação UVC e a mutágenos químicos

Um novo mutante isolado da levedura Saccharomyces cerevisiae, sensível à fotoativação de psoralenos mono- e bi-funcionais, à UVC e ao MNNG, complementou o fenótipo de sensibilidade à fotoadição de psoralenos conferido pelas mutações pso1 a pso7 e assim foi chamado pso8-1. O duplo mutante pso8-1 rad4-4 foi altamente sensível à UVC, indicando assim uma interação sinergística dos dois mutantes alelos. A clonagem molecular pela complementação do fenótipo de sensibilidade à radiação UVC do mutante pso8-1 e estudos genéticos revelaram que pso8-1 é alelo ao gene RAD6. O produto do gene RAD6/UBC2 é uma enzima conjugada à ubiquitina envolvida em reparação de DNA, esporulação, recombinação, indução de mutagênese, degradação de proteínas, genes silenciosos, transposição Ty1. Enquanto o mutante pso8-1 apresenta um fenótipo mutador espontâneo e possui baixa mutabilidade induzida a mutágenos, a esporulação de diplóides homoalélicas mostrou eficiência próxima à da linhagem selvagem. A análise da seqüência do mutante alelo mostrou que pso8-1 contém uma nova, e até agora desconhecida, transição T→C no nucleotídeo 191, levando à substituição de uma prolina altamente conservada por uma leucina na posição 64 (Rad6-[P64L]) que pode ter severas conseqüências na estrutura terciária da proteína mutada, e conseqüente ligação a pRad18.

Hidrogenação de catalítica de acetileno sobre catalisadores de paládio e catalisadores bimetálicos de paládio e prata

Nesse estudo, prepararam-se catalisadores de paládio e catalisadores de paládio e prata que foram avaliados, juntamente com o catalisador comercial G58H para a reação de hidrogenação seletiva de acetileno. Foram preparados materiais com 0,03% e 0,1% de Pd depositados sobre α-Al2O3 e α-Al2O3 impregnada com 0,2% de Ag. A superfície dos catalisadores foi analisada por quimissorção de CO, TPR de H2 e medidas de área superficial pelo método BET. Os catalisadores de paládio e paládio/prata mostraram-se bastante ativos e seletivos para a reação de hidrogenação de acetileno. Os resultados dos ensaios de quimissorção de CO indicaram que os catalisadores monometálicos preparados a partir do precursor Pd(NO3)2 possuem maior dispersão e que esta diminui com o aumento do conteúdo metálico. A adição da prata diminui a dispersão do paládio nos catalisadores bimetálicos. As análises de redução a temperatura programada (TPR) indicaram que os catalisadores monometálicos de paládio preparados a partir do precursor Pd(NO3)2 apresentam um número maior de espécies redutíveis. No caso dos catalisadores bimetálicos, as análises de TPR permitiram a identificação de fases metálicas onde o paládio interage com a prata. prata. As medidas de área superficial sugeriram que a adição da fase metálica reduz a área superficial dos catalisadores. Os ensaios de atividade catalítica mostraram que os catalisadores mono e bimetálicos preparados a partir do precursor Pd(NO3)2 são mais ativos que os preparados a partir do complexo [Pd(acac)2]. A atividade aumenta com percentual de paládio, porém, as menores seletividades foram obtidas para os catalisadores com maior percentual de paládio. Os catalisadores bimetálicos de Pd/Ag mostraram-se menos ativos que os monometálicos de paládio, entretanto foram mais seletivos na formação de etileno.