Oxygen-dependence of metabolic rate in the muscles of craniates

We present evidence that oxygen consumption (V_O2 ) is oxygen partial pressure (P_O2) dependent in striated muscles and P_O2 -independent in the vasculature in representatives of three craniate taxa: two teleost fish, a hagfish and a rat. Blood vessel V_O2 displayed varying degrees of independence in a P_O2 range of 15–95 mmHg, while V_O2 by striated muscle tissue slices from all species related linearly to P_O2 between 0 and 125 mmHg, despite V_O2 rates varying greatly between species and muscle type. In salmon red muscle, lactate concentrations fell in slices incubated at a P_O2 of either 30 or 100 mmHg, suggesting aerobic rather than anaerobic metabolism. Consistent with this finding, potential energy, a proxy of ATP turnover, was P_O2 -dependent. Our data suggest that the reduction in V_O2 with falling P_O2 results in a decrease in ATP demand, suggesting that the hypoxic signal is sensed and cellular changes effected. Viability and diffusion limitation of the preparations were investigated using salmon cardiac and skeletal muscles. Following the initial P_O2 depletion, reoxygenation of the Ringer bathing salmon cardiac muscle resulted in V_O2^s that was unchanged from the first run. V_O2 increased in all muscles uncoupled with p-trifluoromethoxylphenyl-hydrazone (FCCP) and 2,4-dinitrophenol (DNP). Mitochondrial succinate dehydrogenase activity, quantified by reduction of 3-(4,5-dimethylthiazol)-2,5-diphenyl-2H-tetrazolium bromide (MTT) to formazan, was constant over the course of the experiment. These three findings indicate that the tissues remained viable over time and ruled out diffusion-limitation as a constraint on V_O2.

Oxygen consumption and blood flow distribution in perfused skeletal muscle of chinook salmon

An isolated, perfused salmon tail preparation showed oxyconformance at low oxygen delivery rates. Addition of pig red blood cells to the perfusing solution at a haematocrit of 5 or 10% allowed the tail tissues to oxyregulate. Below ca. 60 ml O₂ kg⁻¹ h⁻¹ of oxygen delivery (DO₂), VO₂ was delivery dependent. Above this value additional oxygen delivery did not increase VO₂ of resting muscle above ca. 35 ml O₂ kg⁻¹ h⁻¹. Following electrical stimulation, VO₂ increased to ca. 65 ml O₂ kg⁻¹ h⁻¹, with a critical DO₂ of ca. 150 ml O₂ kg⁻¹ h⁻¹. Dorsal aortic pressure fell to 69% of the pre-stimulation value after 5 min of stimulation and to 54% after 10 min. Microspheres were used to determine blood flow distribution (BFD) to red (RM) and white muscle (WM) within the perfused myotome. Mass specific BFD ratio at rest was found to be 4.03 ± 0.49 (RM:WM). After 5 min of electrical stimulation the ratio did not change. Perfusion with saline containing the tetrazolium salt 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) revealed significantly more mitochondrial activity in RM. Formazan production from MTT was directly proportional to time of perfusion in both red and WM. The mitochondrial activity ratio (RM:WM) did not change over 90 min of perfusion.

Influence of ionic liquids on the electrical conductivity and morphology of PEDOT:PSS films

Small-molecule nonvolatile additives based on ionic liquids (IL) as electrical conductivity enhancer in Poly(3,4-ethylenedioxythiophene): poly(styrenesulfonate) (PEDOT:PSS) was studied. Ionic liquids were investigated in the synthesis of self-assembled, highly organized hybrid nanostructures due to their ability as supramolecular solvents. Different percentage of five ionic liquids, such as 1-butyl-3-methylimidazolium tetrafluoroborate (bmim) F 4 and 1-butyl-3-methylimidazolium bromide (bmim)Br were added to a PEDOT:PSScommercial dispersion. Films of pure PEDOT:PSS showed an average conductivity of 14 S cm-1, which corresponded to the value range given by the supplier. AFM images showed that IL induced the formation of a three-dimensional conducting network with smaller PEDOT domains. The ionic character of the films was significantly increased because of the presence of ionic liquids, which can be used effectively in optoelectronic devices.

Structure retention in cross-linked poly(ethylene glycol) diacrylate hydrogel templated from a hexagonal lyotropic liquid crystal by controlling the surface tension

Retaining hexagonal lyotropic liquid crystal (LLC) structures in polymers after surfactant removal and drying is particularly challenging, as the surface tension existing during the drying processes tends to change the morphology. In this study, cross-linked poly(ethylene glycol) diacrylate (PEGDA) hydrogels were prepared in LLC hexagonal phases formed from a dodecyltrimethylammonium bromide (DTAB)/water system. The retention of the hexagonal LLC structures was examined by controlling the surface tension. Polarized light microscopy, X-ray diffraction and small angle X-ray scattering results indicate that the hexagonal LLC structure was successfully formed before polymerization and well retained after polymerization and after surfactant removal when the surface tension forces remained neutral. Controlling the surface tension during the drying process can retain the nanostructures templated from lyotropic liquid crystals which will result in the formation of materials with desired nanostructures.

Preparation of hemicyanine dye encapsulated silica particles and their application in cotton fabrics

Trans-4- [p- (N, N-Die (2-hydroxyethyl)) styryl] -N- ethyl pyridinium bromide (DHEASPBr-C2), a hemicyanine fluorescent dye, was encapsulated into silica nanoparticles by co-hydrolysis and co-condensation of organosilanes in the presence of the dye. The dye containing silica nanoparticles were applied onto cotton fabrics. Scanning electron microscopy (SEM), UV–vis spectra, single-photon emission fluorescence spectra and reflectance spectra of the samples were characterized. The SEM results showed that the particle size (ranging from 100-200 nm) and dye encapsulating (1.5-8.1 mg dye per g silica matrix) could be adjusted by the concentration of fluorescent dye and organosilanes. The reflectance of the treated cotton fabrics showed that there were obvious adsorption spectra in 410 - 540 nm and emission spectra in 560 - 700 nm.

Use of fluorescein as a ground water tracer in brackish water aquifers

A drift and pumpback experiment was conducted in a brackish water sandfill. The sandfill was reclaimed from the sea in the eastern part of Singapore and contains sands with low organic and clay/silt contents. The high salinity in the ground water precludes the use of chloride and bromide as tracers in such an environment, and a field experiment was conducted to assess the viability of using fluorescein as a tracer in brackish water aquifers. Nitrate was used as a second tracer to serve as a check. Initial laboratory studies showed that fluorescence was unaffected over the range of electrical conductivity and pH of the ground water. Results from the field experiment show that fluorescein appears to behave conservatively.

Homogeneous isolation of nanocelluloses by controlling the shearing force and pressure in microenvironment

Nanocelluloses were prepared from sugarcane bagasse celluloses by dynamic high pressure microfluidization (DHPM), aiming at achieving a homogeneous isolation through the controlling of shearing force and pressure within a microenvironment. In the DHPM process, the homogeneous cellulose solution passed through chambers at a higher pressure in fewer cycles, compared with the high pressure homogenization (HPH) process. X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS) demonstrated that entangled network structures of celluloses were well dispersed in the microenvironment, which provided proper shearing forces and pressure to fracture the hydrogen bonds. Gel permeation chromatography (GPC), CP/MAS 13C NMR and Fourier transform infrared spectroscopy (FT-IR) measurements suggested that intra-molecular hydrogen bonds were maintained. These nanocelluloses of smaller particle size, good dispersion and lower thermal stability will have great potential to be applied in electronics devices, electrochemistry, medicine, and package and printing industry. © 2014 Elsevier Ltd.

A new insight into growth mechanism and kinetics of mesoporous silica nanoparticles by in situ small angle x-ray scattering

The growth mechanism and kinetics of mesoporous silica nanoparticles (MSNs) were investigated for the first time by using a synchrotron time-resolved small-angle X-ray scattering (SAXS) analysis. The synchrotron SAXS offers unsurpassed time resolution and the ability to detect structural changes of nanometer sized objects, which are beneficial for the understanding of the growth mechanism of small MSNs (∼20 nm). The Porod invariant was used to quantify the conversion of tetraethyl orthosilicate (TEOS) in silica during MSN formation, and the growth kinetics were investigated at different solution pH and temperature through calculating the scattering invariant as a function of reaction time. The growth of MSNs was found to be accelerated at high temperature and high pH, resulting in a higher rate of silica formation. Modeling SAXS data of micelles, where a well-defined electrostatic interaction is assumed, determines the size and shape of hexadecyltrimethylammonium bromide (CTAB) micelles before and after the addition of TEOS. The results suggested that the micelle size increases and the micelle shape changes from ellipsoid to spherical, which might be attributed to the solubilization of TEOS in the hydrophobic core of CTAB micelles. A new "swelling-shrinking" mechanism is proposed. The mechanism provides new insights into understanding MSN growth for the formation of functional mesoporous materials exhibiting controlled morphologies. The SAXS analyses were correlated to the structure of CTAB micelles and chemical reaction of TEOS. This study has provided critical information to an understanding of the growth kinetics and mechanism of MSNs.

Solubilizacao e interacao de pigmentos com solventes organicos e agentes tenso-ativos

A Mg e Mn-Ftalocianina (Mg e Mn-Pc) foram solubilizados à 25°C em dimetilsulfóxido (DMSO); N, N-dimetilacetamida (DMA); N,N-dimetilformamida (DMF); N-metil-formamida, formamida, piridina, o-diclorobenzeno, monoclorobenzeno, tolueno, metanol, etanol, propanol-1, propanol-2, butanol-1 e octanol-1. Alguns valores representativos obtidos para o logarítimo da absortividade molar (E) da Mn-Pc, são os seguintes: o-diclorobenzeno (E = 4,94); DMSO (E = 4,39); octanol-1 (E = 3,90). Valores correspondentes para Mg-Pc são: o-diclarobenzeno (E = 4,93); DMSO (E = 5,22) e Octanol-1 ( E = 5,06). Em função de interação com solventes, pode-se classificar a Mg-Pc como um indicador básico e a Mn-Pc como indicador ácido. Os pigmentos Mg e Mn-Pc foram também solubillzados em soluções aquosas contendo vários surfatantes à 25°C. A Mg-Pc apresentou solubilidade significativa em água contendo brometo de cetiltrimetilamônio (CTAB), Brij-35, cloreto de cetilpiridinio (CPC1), brometo de cetilpiridínio (CPBr,) Triton X-100, cloreto de metildodecilbenziltrimetilamônio, brometo de cetildimetiletilamõnio e brometo de laurilisoquinolínio. A Mn-Pc foi solúvel em soluções aquosas de Brij-35 e Triton X-100. Em função de sua interação com surfatantes a Mg-Pc é classificada como corante catiônico e a Mn-Pc como corante aniônico. O corante comercial quinóide Oil Blue A [1,4-di(isopropilarnina)-antraquinona - 9,10 foi solubilizado à 25°C em DMF, DMSO, DMA, monoclorobenzeno, benzeno, tolueno, piridina, metanol, etanol, propanol-1, propanol-2, butanol-1 e octanol-1. Foi também solubilizado em soluções aquosas de surfatantes, tais como sódio lauril-sulfato (NaLS), cloreto de cetiltrimetilamônio (CTAB), brometo de cetildimetiletilamônio, Triton X-100, cloreto de cetilpiridínio (CPCl), Brij-35, cloreto de rnetildodecilbenziltrimetilamônio e brometo de laurilisoquinolínio. Em função de suas interações com os solventes o corante é um indicador ácido-básico pouco sensível e em função de sua interação com surfatantes é um corante catiônico. 0s resultados experimentais apresentam importância teórica e prática considerando sistemas que envolvem armazenamento e transferência de energia, compostos porfirínicos, fotossíntese, fotocondutores, coletores solares, semi-condutores e processos de embelezamento e proteção de superficies de vários materiais.

Estudo da influência da variabilidade em genes de apolipoproteínas sobre níveis lipídicos e parâmetros de massa e gordura corporal na população de Porto Alegre

Apolipoproteínas constituem a parte proteica das lipoproteínas e de uma maneira geral desempenham papéis como proporcionar estabilidade estrutural, solubilizar lipídeos altamente hidrofóbicos, servir como ligantes a receptores ou agir como co-fatores para enzimas envolvidas no metabolismo. Diversos estudos têm mostrado que a variabilidade dos genes que codificam estas proteínas podem influenciar os níveis lipídicos em diversas populações. A variabilidade da apo A-IV também foi associada com variáveis antropométricas. Nesta investigação foram analisados 8 RFLPs nos genes das apolipoproteínas C-I (HpaI), C-II (AvaII), C-III (SacI, FokI e MspI) e A-IV (XbaI, HinfI e PvuII). A amostra foi composta por 391 indivíduos caucasóides de Porto Alegre (RS) e dados sobre hábitos de vida, dosagens lipídicas e medidas antropométricas foram obtidas para cada indivíduo. Os fragmentos de interesse de cada gene foram amplificados por PCR e os genótipos foram identificados por eletroforese em géis de agarose ou poliacrilamida corados com brometo de etídio.

O polimorfismo da apolipoproteina e em três grupos étnicos e suas influências sobre os níveis lipídicos e a doença de Alzheimer

O gene da apolipoproteina E (APOE) possui três alelos com freqüências polimórficas. Esta apolipoproteína possui um importante papel no metabolismo de lipídeos, crescimento e regeneração neuronal, e parece estar relacionada com a doença de Alzheimer. No entanto, a magnitude destas influências difere de acordo com a população estudada, sugerindo uma interação genótipo/ambiente. No presente trabalho, foram estudadas seis tribos indígenas sul-americanas (n=186), 100 negróides e 466 caucasóides de Porto Alegre. Destes últimos, 343 foram investigados quanto à associação com níveis lipídicos e 23 quanto à associação com doença de Alzheimer. Todas as amostras foram amplificadas pela reação em cadeia da polimerase (PCR) e clivadas com a enzima de restrição Hha I. Os genótipos foram identificados após separação dos fragmentos de restrição por eletroforese em gel de agarose a 4% corado com brometo de etídeo. O presente estudo teve os seguintes objetivos específicos: 1)Determinar as freqüências gênicas e genotípicas da APOE nas populações negróides e caucasóides de Porto Alegre e de seis tribos indígenas da América do Sul; 2)Verificar se as associações entre os alelos da APOE e lipídeos séricos descritas em caucasóides também ocorrem em populações indígenas brasileiras; 3)Investigar a influência do polimorfismo do gene APOE em pacientes com hipercolesterolemia e hipertrigliceridemia, bem como em indivíduos normais da população de Porto Alegre e 4)Determinar a distribuição dos alelos da APOE em uma amostra de pacientes com Doença de Alzheimer.

Estimulação colinérgica com piridostigmina reduz arritmia ventricular e aumenta a variabilidade da frequência cardíaca em pacientes com insuficiência cardíaca

INTRODUÇÃO. O aumento da densidade de arritmia ventricular e a redução da variabilidade da freqüência cardíaca estão associados com risco de morte súbita e mortalidade total em insuficiência cardíaca. A inibição colinesterásica com brometo de piridostigmina (PIR) aumenta a variabilidade da freqüência de pessoas normais, porém seu efeito em pacientes com insuficiência cardíaca é desconhecido. OBJETIVOS. Testar a hipótese de que a administração a curto prazo de piridostigmina reduz a densidade de arritmia ventricular e aumenta a variabilidade da freqüência cardíaca em pacientes com insuficiência cardíaca. MÉTODOS. Pacientes com insuficiência cardíaca e em ritmo sinusal participaram de um estudo duplo-cego, cruzado, randomizado para placebo e piridostigmina (30mg VO de 8 em 8 horas por 2 dias). Monitorização eletrocardiográfica ambulatorial de 24 horas foi realizada para análise de arritmia e para avaliação dos índices do domínio do tempo da variabilidade da freqüência cardíaca. Pacientes foram separados em 2 grupos, de acordo com a densidade de arritmia ventricular. O grupo Arritmia (n = 11) incluiu pacientes com mais de 10 extrassístoles ventriculares por hora (ESV/h), e o grupo Variabilidade da Freqüêcia Cardíaca (n = 12) incluiu pacientes com um número de ESVs em 24 horas que não excedia 1 % do número total de intervalos RR. RESULTADOS. No grupo Arritmia, PIR resultou em uma redução de 65% no número de extrassístoles ventriculares (Placebo 266 + 56 ESV/h vs. PIR 173 + 49 ESV/h; p = 0,03). No grupo da Variabilidade da Freqüência Cardíaca, a administração de PIR resultou em um aumento do intervalo RR médio (Placebo 733 + 22 ms vs PIR 790 + 33 ms; p = 0,01), e nos índices do domínio do tempo da variabilidade da freqüência cardíaca PNN50 (Placebo 3 + 1,1 % vs PIR 6 + 1,6 %; p = 0,03) e RMSSD (Placebo 21 + 2 vs PIR 27 + 3; p = 0,008). CONCLUSÃO. Em pacientes com insuficiência cardíaca, PIR reduziu a densidade de arritmia ventricular e aumentou a VFC, provavelmente por seu efeito colinomimético. Estudos a longo prazo com PIR em insuficiência cardíaca devem ser realizados.

Estudos comparativos do ciclo de regeneração de diferentes tipos de silicoaluminofosfatos

Different types of heterogeneous catalysts of the silicoaluminophosphate type, (SAPO-5, SAPO-11, SAPO-31, SAPO-34 and SAPO-41), molecular sieves with a: AFI, AEL, ATO, CHA and AFO structure, respectively, were synthesized through the hydrothermal method. Using sources such as hydrated alumina (pseudobohemita), phosphoric acid, silica gel, water, as well as, different types of organic structural templates, such as: cetyltrimethylammonium bromide (CTMABr), di-isopropylamine (DIPA), di-n- propylamine (DNPA) and tetraethylammonium hydroxide (TEOS), for the respective samples. During the preparation of the silicoaluminophosphates, the crystallization process of the samples occurred at a temperature of approximately 200 ° C, ranging through periods of 18-72 h, when it was possible to obtain pure phases for the SAPOs. The materials were furthermore washed with deionized water, dried and calcined to remove the molecules of the templates. Subsequently the samples were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), absorption spectroscopy in the infrared region (FT-IR), specific surface area and thermal analysis via TG/DTG. The acidic properties were determined using adsorption of n-butylamine followed by programmed termodessorption. These methods revealed that the SAPO samples showed a typically weak to moderate acidity. However, a small amount of strong acid sites was also detected. The deactivation of the catalysts was conducted by artificially coking the samples, followed by n-hexane cracking reactions in a fixed bed with a continuous flow micro-reactor coupled on line to a gas chromatograph. The main products obtained were: ethane, propane, isobutene, n-butane, n-pentane and isopentane. The Vyazovkin (model-free) kinetics method was used to determine the catalysts regeneration and removal of the coke

Renal and antibacterial effects induced by myotoxin I and II isolated from Bothrops jararacussu venom

Bothrops jararacussu myotoxin I (BthTx-I; Lys 49) and II (BthTX-II; Asp 49) were purified by ion-exchange chromatography and reverse phase HPLC. In this work we used the isolated perfused rat kidney method to evaluate the renal effects of B. jararacussu myotoxins I (Lys49 PLA(2)) and II (Asp49 PLA(2)) and their possible blockage by indomethacin. BthTX-1 (5 mu g/ml) and BthTX-II (5 mu g/ml) increased perfusion pressure (PP; ct(120) = 110.28+/-3.70 mmHg; BthTX I = 171.28+/-6.30* mmHg; BthTX II = 175.50+/-7.20* mmHg), renal vascular resistance (RVR; ct(120) = 5.49+/-0.54 mmHg/ml.g(-1) min(-1); BthTX I = 8.62+/-0.37* mmHg/ml g(-1) min(-1); BthTX II=8.9+/-0.36* mmHg/ml g(-1) min(-1)), urinary flow (UF; ct(120)= 0.14+/-0.01 ml g(-1) min(-1); BthTX I=0.32+/-0.05* ml g(-1) min(-1); BthTX II=0.37+/-0.01* ml g(-1) min(-1)) and glomerular filtration rate (GFR; ct(120)=0.72+/-0.10 ml g(-1) min(-1); BthTX I=0.85+/-0.13* ml g(-1) min(-1); BthTX II=1.22+/-0.28* ml g(-1) min(-1)). In contrast decreased the percent of sodium tubular transport (%TNa+; ct(120)=79,76+/-0.56; BthTX I=62.23+/-4.12*; BthTX II=70.96+/-2.93*) and percent of potassium tubular transport (%TK+;ct(120)=66.80+/-3.69; BthTX I=55.76+/-5.57*; BthTX II=50.86+/-6.16*). Indomethacin antagonized the vascular, glomerular and tubular effects promoted by BthTX I and it's partially blocked the effects of BthTX II. In this work also evaluated the antibacterial effects of BthTx-I and BthTx-II against Xanthomonas axonopodis. pv. passiflorae (Gram-negative bacteria) and we observed that both PLA2 showed antibacterial activity. Also we observed that proteins Also we observed that proteins chemically modified with 4-bromophenacyl bromide (rho-BPB) decrease significantly the antibacterial effect of both PLA(2). In conclusion, BthTx I and BthTX II caused renal alteration and presented activity antimicrobial. The indomethacin was able to antagonize totally the renal effects induced by BthTx I and partially the effects promoted by BthTx II, suggesting involvement of inflammatory mediators in the renal effects caused by myotoxins. In the other hand, other effects could be independently of the enzymatic activity of the BthTX II and the C-terminal domain could be involved in both effects promoted for PLA(2). (C) 2005 Elsevier Ltd. All rights reserved.

Structure-function relationship of new crotamine isoform from the Crotalus durissus cascavella

In this work we isolated a novel crotamine like protein from the Crotalus durissus cascavella venom by combination of molecular exclusion and analytical reverse phase HPLC. Its primary structure was:YKRCHKKGGHCFPKEKICLPPSSDLGKMDCRWKRK-CCKKGS GK. This protein showed a molecular mass of 4892.89 da that was determined by Matrix Assisted Laser Desorption Ionization Time-of-flight (MALDI-TOF) mass spectrometry. The approximately pI value of this protein was determined in 9.9 by two-dimensional electrophoresis. This crotamine-like protein isolated here and that named as Cro 2 produced skeletal muscle spasm and spastic paralysis in mice similarly to other crotamines like proteins. Cro 2 did not modify the insulin secretion at low glucose concentration (2.8 and 5.6 mM), but at high glucose concentration (16.7 mM) we observed an insulin secretion increasing of 2.7-3.0-fold than to control. The Na+ channel antagonist tetrodoxin (6 mM) decreased glucose and Cro 2-induced insulin secretion. These results suggested that Na+ channel are involved in the insulin secretion. In this article, we also purified some peptide fragment from the treatment of reduced and carboxymethylated Cro 2 (RC-Cro 2) with cyanogen bromide and protease V8 from Staphylococcus aureus. The isolated pancreatic beta-cells were then treated with peptides only at high glucose concentration (16.7 mM), in this condition only two peptides induced insulin secretion. The amino acid sequence homology analysis of the whole crotamine as well as the biologically-active peptide allowed determining the consensus region of the biologically-active crotamine responsible for insulin secretion was KGGHCFPKE and DCRWKWKCCKKGSG.