Endogenous endothelin-1 limits exercise-induced vasodilation in hypertensive humans

Essential hypertension is a common disorder, associated with increased endothelin-l-mediated vasoconstrictor tone at rest. We hypothesized that increased vasoconstrictor activity of endothelin-1 might explain why the normal decrease in peripheral vascular resistance in response to exercise is attenuated in hypertensive patients. Therefore, we investigated the effect of endothelin A (ETA) receptor blockade on the vasodilator response to handgrip exercise. Forearm blood flow responses to handgrip exercise (15%, 30%, and 45% of maximum voluntary contraction) were assessed in hypertensive patients and matched normotensive subjects, before and after intra-arterial infusions of the ETA receptor antagonist BQ-123; a control dilator, hydralazine; and placebo (saline). Preinfusion (baseline) vasodilation in response to exercise was significantly attenuated at each workload in hypertensive patients compared with normotensive subjects. Intra-arterial infusions of hydralazine and saline did not increase the vasodilator response to exercise in either hypertensives or normotensives at any workload. The vasodilator response to exercise was markedly enhanced after BQ-123 at the 2 higher workloads in hypertensives (157 +/- 48%, P < 0.01; 203 &PLUSMN; 58%, P < 0.01) but not in normotensives. This suggests that the impaired vasodilator response to exercise in hypertensive patients is, at least in part, a functional limitation caused by endogenous ETA receptor-mediated vasoconstriction. Treatment with endothelin receptor antagonists may, therefore, increase exercise capacity in essential hypertension.

Effects of intravenous injection of Clostridium perfringens type D epsilon toxin in calves

In cattle, a neurological lesion similar to that produced in sheep and goats by Clostridium perfringens type D enterotoxaemia has been reported. However, no causal relationship has been established between this disease and the lesion in cattle. The effects of single and multiple intravenous injections of epsilon toxin in three calves aged 6 months were studied. A further calf was inoculated intravenously with saline solution and used as a control. Epsilon toxin invariably produced neurological signs within 2-60 min of the end of the injection process. Clinical signs consisted of loss of consciousness, recumbency, convulsions, paddling, opisthotonus, hyperaesthesia and dyspnoea. Gross changes consisted of severe acute pulmonary oedema, which was particularly marked in the interlobular septa. The histological lesions consisted of intra-alveolar and interstitial oedema of the lung and variable degrees of perivascular proteinaceous oedema in the internal capsule, thalamus and cerebellar white matter. No clinical or post-mortem changes were observed in the control calf. These results show that calves are susceptible to the intravenous injection of epsilon toxin, and that they can show at least some of the histological lesions produced in sheep and goats by this toxin. (C) 2002 Harcourt Publishers Ltd.

Cold Storage Preservation and Warm Ischaemic Injury to Isolated Arterial Segments: Endothelial Cell Injury

Injury to endothelial calls is thought to be important to the development of the vascular lesion of chronic rejection. It was the aim of this study to develop a semiquantitative method to assess endothelial injury in arterial grafts and to document the injury produced by cold storage preservation and additional warm ischaemia. Twelve- and 24-h cold preservation of rat aortic segments, together with an additional 1 h of warm ischaemia, were assessed. Electron micrographs of representative endothelial cells were scored for cytoplasmic, nuclear and mitochondrial injury. The overall injury score was obtained by addition of the individual scores. Storage for up to 24 h in University of Wisconsin (UW) and Terasaki did not produce any injury. Twenty-four hours of storage in Euro-Collins resulted in endothelial cell death. Injury occurred after 12 h of storage in Ross, Collins and normal saline, and the injury increased following 24 h of storage. One hour of warm ischaemia did not increase the injury. Injury to endothelial cells varies with the preservation solution used and the time of cold storage, so that both the type of solution and the storage time should be taken into account in clinical studies looking at the influence of cold ischaemia time and graft outcome.

Antagonism by NKP608 of substance P-induced plasma protein exudation in a novel preparation of perfused trachea in rats and guinea-pigs in vivo

Objective: To examine whether NKP608, a novel 1-benzoyl-2-benzyl-4-aminopiperidine NK1 receptor antagonist, inhibits substance P (SP)-induced airway plasma protein exudation in vivo. Material: Anaesthetised English shorthair guinea-pigs and Wistar rats. Treatment: Tachykinin peptides were applied topically onto the trachea and antagonists administered intravenously. Methods: Tracheal segments isolated in situ were perfused with saline and plasma-derived protein assayed in the perfusate. Results: SP (1 muM) caused plasma protein exudation, which was abolished by an NK1 antagonist (RP 67580, 1.75 mumol/kg) but unaffected by an NK2 antagonist (SR 48968, 1.75 mumol/kg) indicating the response is NK1-receptor-mediated. This was confirmed with a response to an NK1 agonist ([Sar(9), Met(O-2)(11)]-SP, 1 muM) but none to an NK2 agonist ([betaAla(8)]-neurokinin A(4-10), 1 muM). NKP608 inhibited SP responses with estimated ID50 values (mumol/kg) of 0.0044 (guinea-pigs) and 0.19 (rats). Conclusions: NKP608 is an antagonist in vivo of NK1 receptor-induced tracheal plasma protein exudation and is more potent in guinea-pigs than rats.

Differences in mouse strain influence leukocyte and immunoglobulin phenotype response to Porphyromonas gingivalis

This study examined the nature of the infiltrating cells in Porphyromonas gingivalis-induced lesions and immunoglobulins in the serum samples of BALB/c (H-2(d)), C57BL6 (H-2(b)), DBA/2J (H-2(d)) and CBA/CaH (H-2(k)) mice. Mice were immunized intraperitoneally with P. gingivalis outer membrane antigens or sham-immunized with phosphate-buffered saline followed by subcutaneous challenge with live organisms 1 week after the final immunization. The resulting skin abscesses were excised 7 days later, cryostat sections cut and an immunoperoxidase method used to detect the presence of CD4(+) and CD8(+) T-cell subsets, CD14(+) macrophages and CD19(+) B cells. Peroxidase positive neutrophils and IgG1- and IgG2a-producing plasma cells were also identified. Anti P. gingivalis IgG1 and IgG2a subclass antibodies were determined in serum obtained by cardiac puncture. Very few CD8(+) T cells and CD19(+) B cells were found in any of the lesions. The percentages of CD4(+) cells, CD14(+) cells and neutrophils were similar in lesions of immunized BALB/c and C57BL6 mice, with a trend towards a higher percentage of CD14(+) cells in sham-immunized mice. The percentage of CD14(+) cells was higher than that of CD4(+) cells in immunized compared with sham-immunized DBA/2J mice. The percentages of CD4(+) and CD14(+) cells predominated in immunized CBA/CaH mice and CD4(+) cells in sham-immunized CBA/CaH mice. The percentage of neutrophils in immunized CBA/CaH mice was significantly lower than that of CD14(+) cells and CD4(+) cells in sham-immunized mice. IgG1(+) plasma cells were more dominant than IgG2a(+) cells in immunized BALB/c, C57BL6 and DBA/2J mice, whereas IgG2a(+) plasma cells were more obvious in sham-immunized mice. IgG2a(+) plasma cells were predominant in immunized and sham-immunized CBA/CaH mice. In the serum, specific anti-P. gingivalis IgG2a antibody levels (Th1 response) were higher than IgG1 levels (Th2 response) in sham-immunized CBA/CaH and DBA/2J mice. In immunized BALB/c mice, IgG2a levels were lower than IgG1 levels, while IgG2a levels were higher in immunized C57BL6 mice. In conclusion, this study has shown differences in the proportion of infiltrating leukocytes and in the subclasses of immunoglobulin produced locally and systemically in response to P. gingivalis in different strains of mice, suggesting a degree of genetic control over the response to P. gingivalis.

Rapid colour changes in multilayer reflecting stripes in the paradise whiptail, Pentapodus paradiseus

The Paradise whiptail (Pentapodus paradiseus) has distinct reflective stripes on its head and body. The reflective stripes contain a dense layer of physiologically active iridophores, which act as multilayer reflectors. The wavelengths reflected by these stripes can change from blue to red in 0.25 s. Transmission electron microscopy revealed that the iridophore cells contain plates that are, on average, 51.4 nm thick. This thickness produces a stack, which acts as an ideal quarter-wavelength multilayer reflector (equal optical thickness of plates and spaces) in the blue, but not the red, region of the spectrum. When skin preparations were placed into hyposmotic physiological saline, the peak wavelength of the reflected light shifted towards the longer (red) end of the visible spectrum. Hyperosmotic saline reversed this effect and shifted the peak wavelength towards shorter (blue/UV) wavelengths. Norepinephrine (100 mumol l(-1)) shifted the peak wavelength towards the longer end of the spectrum, while adenosine (100 mumol l(-1)) reversed the effects of norepinephrine. The results from this study show that the wavelength changes are elicited by a change of similar to70 nm in the distance between adjacent plates in the iridophore cells.

Bovine-serum-albumin-containing receptor phase better predicts transdermal absorption parameters for lipophilic compounds

Based on the hypothesis that limited receptor solubility of lipophilic compounds may result in lower observed permeability parameters, the aim of this study was to determine the in vitro human epidermal permeability coefficients and membrane retention of a series of aliphatic alcohols (C1-C10, log p -0.72 to 4.06) using two different receptor solutions (water and 4% bovine serum albumin in phosphate-buffered saline). Aqueous solutions of radiolabeled alcohols were dosed into the stratum corneum side of membranes mounted in side-by-side glass diffusion cells. Appearance of alcohol in the receptor compartment filled with either of the two solutions was monitored over a 7 h period when both stratum corneum (assessed by tape stripping) and the remaining epidermis levels of radioactivity were determined. In a separate study the degree of binding of alcohols to 4% bovine serum albumin was determined. The data showed increased receptor phase solubility in the bovine serum albumin solution and higher permeability coefficients for the more lipophilic alcohols in the series. No changes were seen in the partitioning of the alcohols from the vehicle into either the stratum corneum or tape-stripped epidermis with the two receptor phases; however, a decrease in the amount of the more lipophilic alcohols partitioning into the water receptor phase from the tape-stripped epidermis was observed. We conclude that bovine serum albumin receptor phase allows better estimation of real permeability parameters for lipophilic compounds due to its increased solubility capacity and we question whether permeability parameters for lipophilic solutes from older data sets based on aqueous receptor phases are completely reliable.

Use of a thermodilution catheter to measure core body temperature, central venous pressure and cardiac output during anaesthesia and surgery in the dog

The use of thermodilution and other methods of monitoring in dogs during surgery and critical care was evaluated. Six Greyhounds were anaesthetised and then instrumented by placing a thermodilution catheter into the pulmonary artery via the jugular vein. A catheter in the dorsal pedal artery also permitted direct measurement of arterial pressures. Core body temperature (degreesC) and central venous pressure (mmHg) were measured, while cardiac output (mL/min/kg) and mean arterial pressure (mmHg) were calculated. A mid-line surgical incision was performed and the physiological parameters were monitored for a total of two hours. All physiological parameters generally declined, although significant increases (P<0.05) were noted for cardiac output following surgical incision. Central venous pressure was maintained at approximately 0mmHg by controlling an infusion of sterile saline. Core body temperature decreased from 37.1+/-0.6degreesC (once instrumented) to 36.6+/-0.60degreesC (at the end of the study), despite warming using heating pads. Physiological parameters indicative of patient viability will generally decline during surgery without intervention. This study describes an approach that can be undertaken in veterinary hospitals to accurately monitor vital signs in surgical and critical care patients.

Variação radial da retratibilidade e densidade básica da madeira de Eucalyptus saligna Sm.

A ultra-estrutura e a composição química da madeira, bem como suas propriedades físicas e mecânicas, variam significativamente entre espécies, entre árvores de uma mesma espécie e, mesmo, entre diferentes partes de uma mesma árvore. Com este trabalho objetivou-se o estudo dos parâmetros de retratibilidade e de densidade básica da madeira Eucalyptus saligna, com idade de 16 anos, proveniente de talhões experimentais da EMBRAPA Florestas, de Colombo, Paraná. As amostras foram retiradas à altura do DAP de quatro posições eqüidistantes a partir da medula em direção à periferia, correspondendo a 0, 33, 66 e 100%, com dimensões nominais de 1,0 x 2,0 x 3,0 cm, sendo a última dimensão no sentido longitudinal. Elas foram mantidas em câmara fechada com ventilação, próximo de soluções salinas supersaturadas, com o objetivo de proporcionar diferentes condições de umidade relativa. Uma vez atingidas as distintas condições de umidade de equilíbrio, as amostras foram secas em estufa a 105 ºC e obtidos os dados de retratibilidade e densidade básica da madeira nas posições mencionadas. Constataram-se valores de contração volumétrica mais baixos na região medular, apresentando um acréscimo para as demais posições. Comportamento semelhante foi observado para os coeficientes das contrações lineares nas direções tangencial e radial. O fator anisotrópico foi consideravelmente mais elevado na região medular, decrescendo substancialmente em direção ao alburno. A densidade básica não mostrou sinais efetivos de estabilidade, apesar de mostrar tendência de aumento em direção à periferia do tronco.

Avaliação das propriedades higroscópicas da madeira de Eucalyptus saligna Sm., em diferentes condições de umidade relativa do ar

O estudo da higroscopicidade é indispensável para o entendimento da trabalhabilidade, estabilidade dimensional, resistência mecânica e durabilidade natural da madeira. Neste trabalho objetivou-se a avaliação do teor de equilíbrio higroscópico para diversas condições de umidade relativa do ar, bem como da retratibilidade linear e volumétrica e da densidade básica da madeira de Eucalyptus saligna Sm. A madeira utilizada foi proveniente de árvores com 16 anos de idade, procedentes de talhões experimentais da EMBRAPA Florestas de Colombo, Paraná. Amostras com dimensões de 1,0x2,0x3,0 cm, sendo a última na direção longitudinal, foram colocadas em uma câmara fechada, sob ventilação, próximas de recipientes com soluções salinas supersaturadas, a fim de atingir determinada condição preestabelecida de teor de equilíbrio higroscópico. Após o equilíbrio da umidade da madeira nas distintas condições de umidade relativa, as amostras foram secas em estufa, para posterior avaliação. Os dados relativos à umidade de equilíbrio ajustaram-se muito bem às condições de umidade relativa adotadas neste estudo, tendo sido possível estimar com grande precisão o teor de equilíbrio higroscópico, para a faixa de aproximadamente 20 até 100% de umidade relativa. A madeira em estudo apresentou dados de retratibilidade bastante elevados, se comparados aos de outras da mesma faixa de densidade. Apesar dos elevados coeficientes de contração, o fator anisotrópico ou relação T/R mostrou-se próximo daquele encontrado na grande maioria das madeiras comerciais brasileiras. Verificaram-se ainda coeficientes de contrações mais suaves nos teores de umidade abaixo de 17%.

Qualidade fisiológica de sementes e diversidade genética de maracujazeiros cultivados em diferentes altitudes no Espírito Santo

O maracujazeiro pertence à família Passifloraceae e ao gênero Passiflora que é o mais importante economicamente. Altitudes entre 100 a 900 m são indicadas para o plantio do maracujazeiro e estudos sobre localizações geográficas distintas possibilitam expressões do genótipo, influenciadas pelas condições ambientais. O gradiente altitudinal influencia a distribuição da variação genética dentro e entre populações de plantas e a diversidade genética muda com a altitude. O presente trabalho teve como objetivo avaliar a qualidade fisiológica de sementes e a diversidade genética de maracujazeiros cultivados em diferentes altitudes do Espírito Santo. Para avaliação da qualidade fisiológica das sementes, os frutos de Passiflora spp. maduros foram colhidos em pomares localizados em altitudes baixa (0-100 m), média (>100 até 600 m) e alta (>600 m) de diferentes municípios do Espírito Santo. Os tratamentos pré-germinativos nas sementes foram: T1- escarificação física, feita manualmente com lixa d´água nº 120; T2- tratamento com ácido giberélico (GA3) na concentração de 500 mg L-1 com embebição por 24 horas e T3- sementes sem escarificação realizados nas sementes em laboratório e em casa de vegetação. Foi avaliado o envelhecimento acelerado tradicional, envelhecimento acelerado com saturação salina e deterioração controlada em sementes de maracujá amarelo sem escarificação localizado em alta altitude e as condições que apresentaram menor deterioração das sementes para aplicação às demais espécies e altitudes com os respectivos tratamentos pré-germinativos que apresentaram maiores valores de germinação e vigor em laboratório e em casa de vegetação. Assim, para as sementes do maracujá amarelo utilizou-se o tratamento sem escarificação, para sementes de maracujá roxo, a escarificação física e para as sementes de maracujá doce, o tratamento com ácido giberélico. O teste de envelhecimento acelerado com saturação salina a 43 ºC por 72 horas e deterioração controlada a 25% de umidade expostas por 24 horas diferencia as espécies nas diferentes altitudes. Sementes de maracujá amarelo e sementes localizadas em alta altitude apresentam qualidade fisiológica superior. Para a avaliação da diversidade genética foram utilizadas folhas jovens de cinco plantas matrizes de Passiflora edulis Sims f. flavicarpa Degener, P. edulis Sims e P. alata Curtis cultivadas em três altitudes (baixa, média e alta) do Espírito Santo. Para SSR foi encontrado baixo número de alelos, alta heterozigosidade esperada e altos valores de PIC e para a análise ISSR detectou um elevado número de bandas por primer e alto polimorfismo. Há maior similaridade genética entre P. edulis f. flavicarpa Deg. e P. edulis. Passiflora alata apresenta maior distância genética em relação às espécies. As populações de baixa altitude se diferenciam das demais independente da espécie e do marcador utilizado.

Efeitos do isolamento social neonatal, da fluoxetina e do lactato desódio sobre o pânico experimental induzido por cianeto de potássio ou estimulação da matéria cinzenta periaquedutal em ratos

Os ataques de pânico (AP) podem ser precipitados pela inalação de dióxido de carbono (CO2 5%) ou pela infusão de lactato de sódio 0,5 mol/L (LAC) em pacientes predispostos, mas não nos indivíduos sadios ou pacientes com outros transtornos psiquiátricos. Estas e outras observações sugeriram que os AP sejam "alarmes falsos de sufocamento". O TP também caracteriza-se pela alta comorbidade com ansiedade de separação da infância (ASI). De fato, a ASI tem sido considerada como um fator predisponente tanto do pânico como da resistência aos panicolíticos. Estudos pré-clínicos do nosso laboratório mostraram, por outro lado, que a fluoxetina (FLX) atenua o pânico experimental à estimulação elétrica da matéria cinzenta periaquedutal dorsal (MCPAd) e à injeção de cianeto de potássio (KCN) em doses e regimes similares aos empregados na clínica. Adicionalmente, estes estudos mostraram que o isolamento social neonatal (ISN), um modelo de ASI, abole os efeitos panicolíticos da fluoxetina (1-2 mg.kg-1.dia-1, 21 dias) no modelo de pânico por estimulação da MCPAd. Portanto, o presente trabalho avaliou os efeitos da administração de 4 mg/kg de FLX (Estudo-1) e da infusão de uma solução 0,5 mol/L de LAC (Estudo-2) sobre as respostas de pânico à estimulação da MCPAd ou à injeção de KCN, respectivamente, em ratos submetidos ao isolamento social neonatal tanto efetivo (ISN) quanto fictício (ISF). No Estudo-1, ratos Wistar machos adultos submetidos ao ISN (3 h diárias) ou ao ISF (somente manipulação), ao longo da lactação, foram implantados com eletrodos na MCPAd e tratados com salina (0,9%, SAL) ou FLX (4 mg.kg-1.dia-1) ao longo de 21 dias. Nos ratos tratados com SAL, enquanto os limiares das respostas de pânico mantiveram-se praticamente inalterados no grupo ISF, eles foram progressivamente aumentados no grupo ISN. Os ratos tratados com FLX4 apresentaram limiares mais elevados que aqueles dos ratos tratados com SAL. A comparação aos limiares basais mostrou que a FLX4 teve efeitos diferenciados nos grupos ISF e ISN, aumentando ou reduzindo seus limiares, respectivamente. Embora não tenhamos observado diferenças dos efeitos da FLX4 sobre os limiares de pânico dos grupos ISN e ISF, as variações percentuais em relação à sessão de triagem indicam que a FLX4 teve efeitos até mesmo facilitadores. Estes resultados estendem observações anteriores da ausência de efeitos de doses menores de FLX nos ratos submetidos ao ISN. Por sua vez, o Estudo-2 mostrou que a infusão endovenosa de uma concentração clinicamente eficaz de LAC (0,5 mol/L) não tem efeito algum sobre o pânico induzido pelo KCN em ratos tanto virgens quanto submetidos ao ISF ou ISN. Embora os últimos resultados sugiram que os AP ao KCN (em ratos) e LAC (em humanos) envolvam mecanismos distintos, a conclusão definitiva requer experimentos adicionais com concentrações maiores de LAC.

Thermal and hydrolytic degradation of electrospun fish gelatin membranes

The thermal and hydrolytic degradation of electrospun gelatin membranes cross-linked with glutaraldehyde in vapor phase has been studied. In vitro degradation of gelatin membranes was evaluated in phosphate buffer saline solution at 37 ºC. After 15 days under these conditions, a weight loss of 68 % was observed, attributed to solvation and depolymerization of the main polymeric chains. Thermal degradation kinetics of the gelatin raw material and as-spun electrospun membranes showed that the electrospinning processing conditions do not influence polymer degradation. However, for cross-linked samples a decrease in the activation energy was observed, associated with the effect of glutaraldehyde cross-linking reaction in the inter- and intra-molecular hydrogen bonds of the protein. It is also shown that the electrospinning process does not affect the formation of the helical structure of gelatin chains.

Modifying fish gelatin electrospun membranes for biomedical applications: cross- linking and swelling behavior

Development of suitable membranes is a fundamental requisite for tissue and biomedical engineering applications. This work presents fish gelatin random and aligned electrospun membranes cross-linked with glutaraldehyde (GA). It was observed that the fiber average diameter and the morphology is not influenced by the GA exposure time and presents fibers with an average diameter around 250 nm. Moreover, when the gelatin mats are immersed in a phosphate buffered saline solution (PBS), they can retain as much as 12 times its initial weight of solution almost instantaneously, but the material microstructure of the fiber mats changes from the characteristic fibrous to an almost spherical porous structure. Cross-linked gelatin electrospun fiber mats and films showed a water vapor permeability of 1.37 ± 0.02 and 0.13 ± 0.10 (g.mm)/(m2.h.kPa), respectively. Finally, the processing technique and cross-linking process does not inhibit MC-3T3-E1 cell adhesion. Preliminary cell culture results showed good cell adhesion and proliferation in the cross-linked random and aligned gelatin fiber mats.

Hydraulic conductivity in response to exchangeable sodium percentage and solution salt concentration

Hydraulic conductivity is determined in laboratory assays to estimate the flow of water in saturated soils. However, the results of this analysis, when using distilled or deionized water, may not correspond to field conditions in soils with high concentrations of soluble salts. This study therefore set out to determine the hydraulic conductivity in laboratory conditions using solutions of different electrical conductivities in six soils representative of the State of Pernambuco, with the exchangeable sodium percentage adjusted in the range of 5-30%. The results showed an increase in hydraulic conductivity with both decreasing exchangeable sodium percentage and increasing electrical conductivity in the solution. The response to the treatments was more pronounced in soils with higher proportion of more active clays. Determination of hydraulic conductivity in laboratory is routinely performed with deionized or distilled water. However, in salt affected soils, these determinations should be carried out using solutions of electrical conductivity different from 0 dS m-1, with values close to those determined in the saturation extracts.