Short isocapnic hyperoxia affects indices of vascular remodeling and intercellular adhesion molecules in healthy men

In preparation for tracheal intubation during induction of anesthesia, the patient may be ventilated with 100% oxygen. To investigate the impact of acute isocapnic hyperoxia on endothelial activation and vascular remodeling, ten healthy young men (24±3 years) were exposed to 5-min normoxia (21% O2) and 10-min hyperoxia trials (100% O2). During hyperoxia, intercellular adhesion molecules (ICAM-1) (hyperoxia: 4.16±0.85 vs normoxia: 3.51±0.84 ng/mL, P=0.04) and tissue inhibitor matrix metalloproteinase 1 (TIMP-1) (hyperoxia: 8.40±3.84 vs normoxia: 5.73±2.15 pg/mL, P=0.04) increased, whereas matrix metalloproteinase (MMP-9) activity (hyperoxia: 0.53±0.11 vs normoxia: 0.68±0.18 A.U., P=0.03) decreased compared to the normoxia trial. We concluded that even short exposure to 100% oxygen may affect endothelial activation and vascular remodeling.

The efficacy of clove oil as an anaesthetic and in euthanasia procedure for small-sized tropical fishes

Abstract Clove oil is used as a fish anesthetic because it is a natural and inexpensive product with low toxicity risks. The goal of the present study was to determine the appropriate concentration of clove oil for small-sized tropical fish to be used in mark-recapture studies or when individuals are to be sacrificed. We applied three different clove oil concentrations (D1=0.05 mL, D2=0.10 mL and D3=0.20 mL per 500 mL of water) on three small-sized fish species. We found a negative relationship between induction time and treatment for two species (Hyphessobrycon sp.1 and Hemigrammus sp.), while concentration was unrelated to recovery time. Fish body length was positively related to induction time in the D2 treatment for Hemigrammus sp., and negatively for Hyphessobrycon sp.1 in the D1 treatment, but was unrelated to recovery time for three species and treatments. Mortality rates varied across treatments, but higher rates were observed with higher clove oil concentrations. We conclude that 0.05 mL of clove oil per 500 mL of water is the most efficient dose for studies where fish will be released back to their natural habitats, while 0.20 mL of clove oil is recommended for studies that require fish euthanization for further laboratory analyses.

Resumo O óleo de cravo é recomendado como anestésico para peixes por ser produto de origem natural, baixo custo e apresentar poucos riscos de intoxicação. O objetivo deste trabalho foi determinar concentrações adequadas de óleo de cravo para anestesiar ou eutanasiar peixes de pequeno porte em ambiente natural. Foram testadas três concentrações do anestésico (D1=0,05 mL, D2=0,10 mL e D3=0,20 mL) em três espécies de peixes de pequeno. Houve uma relação negativa entre o tempo para a sedação dos indivíduos e a concentração para duas espécies (Hyphessobrycon sp.1 e Hemigrammus sp.), porém não foi encontrada relação entre o tempo para recuperação e as concentrações. Os exemplares maiores de Hemigrammus sp. levaram mais tempo para serem sedados no tratamento D2, já o contrário foi observado para Hyphessobrycon sp.1 no tratamento D1, enquanto que não houve efeito do comprimento no tempo de recuperação das três espécies. A mortalidade dos indivíduos variou entre as três concentrações do anestésico e as maiores taxas de mortalidade ocorreram nas maiores concentrações. Desse modo, a concentração de 0,05 mL é eficiente para estudos que envolvem manuseio e a soltura dos peixes, enquanto que a concentração de 0,20 mL é recomendada em estudos onde os peixes precisam ser sacrificados.

The efficacy of clove oil as an anaesthetic and in euthanasia procedure for small-sized tropical fishes / A eficiência do óleo de cravo como anestésico e em procedimentos de eutanaisa em peixes de pequeno porte

Abstract Clove oil is used as a fish anesthetic because it is a natural and inexpensive product with low toxicity risks. The goal of the present study was to determine the appropriate concentration of clove oil for small-sized tropical fish to be used in mark-recapture studies or when individuals are to be sacrificed. We applied three different clove oil concentrations (D1=0.05 mL, D2=0.10 mL and D3=0.20 mL per 500 mL of water) on three small-sized fish species. We found a negative relationship between induction time and treatment for two species (Hyphessobrycon sp.1 and Hemigrammus sp.), while concentration was unrelated to recovery time. Fish body length was positively related to induction time in the D2 treatment for Hemigrammus sp., and negatively for Hyphessobrycon sp.1 in the D1 treatment, but was unrelated to recovery time for three species and treatments. Mortality rates varied across treatments, but higher rates were observed with higher clove oil concentrations. We conclude that 0.05 mL of clove oil per 500 mL of water is the most efficient dose for studies where fish will be released back to their natural habitats, while 0.20 mL of clove oil is recommended for studies that require fish euthanization for further laboratory analyses.

Resumo O óleo de cravo é recomendado como anestésico para peixes por ser produto de origem natural, baixo custo e apresentar poucos riscos de intoxicação. O objetivo deste trabalho foi determinar concentrações adequadas de óleo de cravo para anestesiar ou eutanasiar peixes de pequeno porte em ambiente natural. Foram testadas três concentrações do anestésico (D1=0,05 mL, D2=0,10 mL e D3=0,20 mL) em três espécies de peixes de pequeno. Houve uma relação negativa entre o tempo para a sedação dos indivíduos e a concentração para duas espécies (Hyphessobrycon sp.1 e Hemigrammus sp.), porém não foi encontrada relação entre o tempo para recuperação e as concentrações. Os exemplares maiores de Hemigrammus sp. levaram mais tempo para serem sedados no tratamento D2, já o contrário foi observado para Hyphessobrycon sp.1 no tratamento D1, enquanto que não houve efeito do comprimento no tempo de recuperação das três espécies. A mortalidade dos indivíduos variou entre as três concentrações do anestésico e as maiores taxas de mortalidade ocorreram nas maiores concentrações. Desse modo, a concentração de 0,05 mL é eficiente para estudos que envolvem manuseio e a soltura dos peixes, enquanto que a concentração de 0,20 mL é recomendada em estudos onde os peixes precisam ser sacrificados.

Quantification of theophylline or paracetamol in milk matrices by high-performance liquid chromatography / 药物分析学报

A simple, accurate and sensitive high-performance liquid chromatography (HPLC) method was developed, validated and applied to the determination of either theophylline or paracetamol in milk-based samples. The method allowed drug quantification in fresh and powdered milk with a relatively short run time of analysis and it was also successfully applied to the quantification of the drugs in solid dosage forms intended for pediatric use. Moreover, the main significant advantages over other published works are the simplicity of the sample preparation, reduced assay time and sample loss. The method meets the International Conference on Harmonization guideline for analytical methods validation regarding specificity,linearity,accuracy,precision, specificity and robustness as required by health authorities and applied by industry while designing and marketing new drug products.The technique encompasses the separation of the analytes with a reverse phase C18column under isocratic conditions and UV detection at 272 nm and 243 nm,respectively,for theophylline and paracetamol. The lower limit of quantification for both drugs was determined as 0.2μg/mL and the between-batch accuracy was 99.7%.This HPLC method allows quantification of theophylline and paracetamol in milk matrices and it can be applied in the design,development and production of milk-based pediatric dosage forms.

Effects of disturbed blood flow during exercise on endothelial function: a time course analysis

This study aimed to examine the time course of endothelial function after a single handgrip exercise session combined with blood flow restriction in healthy young men. Nine participants (28±5.8 years) completed a single session of bilateral dynamic handgrip exercise (20 min with 60% of the maximum voluntary contraction). To induce blood flow restriction, a cuff was placed 2 cm below the antecubital fossa in the experimental arm. This cuff was inflated to 80 mmHg before initiation of exercise and maintained through the duration of the protocol. The experimental arm and control arm were randomly selected for all subjects. Brachial artery flow-mediated dilation (FMD) and blood flow velocity profiles were assessed using Doppler ultrasonography before initiation of the exercise, and at 15 and 60 min after its cessation. Blood flow velocity profiles were also assessed during exercise. There was a significant increase in FMD 15 min after exercise in the control arm compared with before exercise (64.09%±16.59%, P=0.001), but there was no change in the experimental arm (-12.48%±12.64%, P=0.252). FMD values at 15 min post-exercise were significantly higher for the control arm in comparison to the experimental arm (P=0.004). FMD returned to near baseline values at 60 min after exercise, with no significant difference between arms (P=0.424). A single handgrip exercise bout provoked an acute increase in FMD 15 min after exercise, returning to near baseline values at 60 min. This response was blunted by the addition of an inflated pneumatic cuff to the exercising arm.

Avaliação de níveis de glicina+serina em dietas de frangos de corte com reduzido teor de proteína bruta / Evaluation of glycine+serine levels in low crude protein diets for broiler chickens

O experimento foi conduzido em delineamento inteiramente ao acaso para avaliar os efeitos de diferentes níveis de glicina+serina (gli+ser) total em dietas de baixa proteína bruta (PB) sobre o desempenho e características de carcaça de frangos de corte de um a 21 dias de idade. Foram utilizadas 750 aves, distribuídas em cinco tratamentos, cinco repetições e 30 aves por unidade experimental. As dietas utilizadas continham 190g/kg de PB e níveis de gli+ser de 16,7; 19,2; 21,7 e 24,2g/kg; a dieta controle continha 230g/kg de PB. Avaliaram-se os efeitos dos níveis de glicina+serina sobre o desempenho e composição de carcaça. Os dados foram submetidos à análise de variância e os resultados obtidos com as dietas de baixos níveis proteicos com diferentes níveis de gli+ser foram comparados à dieta controle pelo teste de Dunnett. Os níveis de gli+ser apresentaram efeito linear sobre a conversão alimentar, ganho de peso e peso aos 21 dias, sendo o nível de 24,2g/kg de gli+ser com resultado semelhante à dieta controle. A redução proteica aumentou o conteúdo de extrato etéreo na carcaça.(AU)

A completely randomized experimental design was carried out to evaluate the effects of total gly+ser levels in low crude protein diets on performance and body composition of male broiler chickens from 1 to 21 days of age. Seven hundred and fifty broiler chickens were used in each one of the production phases. The birds were randomly allotted to five treatments and five replicates. The diets contained 190g/kg crude protein (CP) and total gly+ser levels of 16.7; 19.2; 21.7 and 24.2g/kg; and a control diet with 230 g/kg CP. The effect of glycine+serine levels on performance and body composition was evaluated. Data were subjected to analysis of variance and the results obtained with the diets of low crude protein levels with different levels of Gly + Ser were compared to the control diet by Dunnett's test. The gly+ser levels improved the feed linearly: weight gain ratio; weight gain and final weight. The broilers fed 24.2g/kg gly+ser diet showed a performance similar to broilers fed control diets. The carcass fat increased with CP reduction.(AU)

Teores de glicina+ serina total em dietas de baixa proteína bruta para de frangos de corte de 22 a 35 dias de idade / Total glycine+serine level in low crude protein diets of broiler chickens from 22 to 35 days of age

O experimento foi realizado em delineamento inteiramente ao acaso para avaliar os efeitos de diferentes teores de glicina + serina (gli+ser) total em dietas de baixa proteína bruta (PB) sobre o desempenho e as características de carcaça de frangos de corte de 22 a 35 dias de idade. Foram utilizadas 750 aves distribuídas em cinco tratamentos, cinco repetições e 30 aves por unidade experimental. As aves foram alimentadas até os 21 dias com uma dieta comum de acordo com as exigências. A partir do 21º dia, as dietas utilizadas continham 17 por cento de PB e níveis de gli+ser de 1,50; 1,75; 2,00 e 2,25 por cento; a dieta-controle continha 21 por cento de PB. O teor de gli+ser teve efeito linear sobre a conversão alimentar e não sobre as demais variáveis de desempenho. Não houve efeito de tratamento sobre os rendimentos de carcaça e cortes. Houve efeito linear decrescente do teor de gli+ser sobre a matéria seca da carcaça.

A completely randomized experimental design was carried out to evaluate the effects of total glycine+serine (gly+ser) levels in low crude protein (CP) diets on performance and body composition of male broiler from 22 to 35 days of age (growing phase). A total of 750 broilers were randomly allotted to five treatments and five replicates of 30 chickens per replication. The birds were fed from 1 to 21 days of age, a common diet formulated to meet bird requirements in all nutrients. From 21 days the diets contained 17 percent CP and gly+ser levels of 1.50; 1.75; 2.00 and 2.25 percent; and a control diet with 21 percentCP. The gly+ser levels showed linear effect on feed: weight gain ratio during the growing phase. There was no effect of gly+ser level on carcass or main carcass part yields. Increasing levels of gly+ser decreased linearly the carcass dry matter content.

Suplementação de betaglucano a dietas de leitões de 21 a 60 dias de idade / Beta-glucon suplementacion in diets for piglets from 21 to 60 days of age

Realizou-se um experimento com 1.500 leitões distribuídos em delineamento experimental de blocos ao acaso com cinco tratamentos: controle e com suplementação de 60, 120, 180 e 240g de betaglucano por tonelada de dieta. Foram analisadas as variáveis ganho de peso diário, peso final, consumo de dieta diário e conversão alimentar nos períodos de 21 a 35, 21 a 49 e 21 a 60 dias de idade. Houve aumento linear significativo do peso final e do ganho de peso diário de leitões suplementados com betaglucano na dieta dos 21 aos 60 dias de idade. A inclusão de 240g/ton. proporcionou aumento no peso final de 800g, o que corresponde ao aumento de 3,2 por cento em relação aos animais do grupo-controle. O ganho de peso diário foi 4,7 por cento mais alto para o grupo de animais tratados com 240g/ton. Não se observou efeito significativo dos tratamentos sobre: consumo diário de dieta, conversão alimentar, atividade da enzima superóxido dismutase nem sobre a resposta imune.

A total of 1,500 piglets were used in a completely randomized experimental block design to study the effects of beta-glucon level (control, 60, 120, 180, and 240g per ton of diet) on daily weight gain, body weight, daily feed intake and feed: weight gain ratio from 21 to 35, 21 to 49 and from 21 to 60 days of age Positive and significant linear effects of beta-gluton on body weight and daily weight gain of piglets from 21 to 60 days of age were observed. The inclusion of 240g of beta-gluton in diet resulted in 800g increase in body weight which corresponds to 3.25 percent increase in body weight and 4.7 percent increase in daily weight gain in comparison to animals of the control group. No effects of beta-gluton supplementation on feed intake, feed: weight gain ratio and superoxide dismutase enzyme activity or animal immune response were observed.

Comparison of different monoclonal antibodies against immunosuppressive proteins of Ascaris suum

The extract of Ascaris suum suppresses the humoral and cellular immune responses to unrelated antigens in the mouse. In order to further characterize the suppressive components of A. suum, we produced specific monoclonal antibodies which can provide an important tool for the identification of these proteins. The A. suum immunosuppressive fractions isolated by gel filtration from an extract of adult worms were used to immunize BALB/c mice. Popliteal lymph node cells taken from the immunized animals were fused with SP2/O myeloma cells and the cloned hybrid cells obtained were screened to determine the specificity of secreted antibodies. Three monoclonal antibodies named MAIP-1, MAIP-2 and MAIP-3 were selected and were shown to react with different epitopes of high molecular weight proteins from the A. suum extract. All antibody molecules have kappa-type light chains but differ in heavy chain isotype. MAIP-1 is a mouse IgM, MAIP-2 is an IgA immunoglobulin and MAIP-3 is an IgG1 immunoglobulin and they recognize the antigen with affinity constants of 1.3 x 10(10) M-1, 7.1 x 10(9) M-1 and 3.8 x 10(7) M-1, respectively. The proteins recognized by these monoclonal antibodies (PAS-1, PAS-2 and PAS-3) were purified from the crude extract by affinity chromatography and injected with ovalbumin in BALB/c mice in order to determine their suppressive activity on heterologous antibody production. It was demonstrated that these three proteins are able to significantly suppress anti-ovalbumin antibody secretion, with PAS-1 being more efficient than the others.

Biomarcador da modificaçäo oxidativa da LDL in vivo / Biomarker LDL's oxidative modification in vivo

As modificaçöes oxidativas da lipoproteína de baixa densidade (LDL) säo consideradas um fator importante para o desenvolvimento da aterosclerose. Estas modificaçöes ocorrem in vivo, originando uma sub-traçäo denominada de LDL, eletronegativa (LDL-). O monitoramento clínico da LDL- é de extrema importância, mas estava sendo limitado pela dificuldade para detecçäo desta partícula em fluídos biológicos. Neste estudo desenvolveu-se novas metodologias para detectar a LDL- no plasma, utilizando-se um anticorpo monoclonal anti-LDL- humana (3D1036) e avaliar a resposta imune humoral relacionada à LDL-. A LDL- plasmática foi analisada através de um ELISA com detecçäo por quimioluminescência com boa sensibilidade (<1,0µg/mL) e precisäo (CVintra=6,44 ñ 1,15 porcento e CVinter=8,59 ñ 3,42 porcento). As análises dos auto-anticorpos anti-LDL- evidenciaram a presença de uma resposta imune específica para LDL- em humanos e em coelhos. A determinaçäo da LDL-, abre novas perspectivas para o monitoramento das modificaçöes oxidativas endógenas da LDL em estudos clínicos e de intervençäo que utilizam um elevado número de amostras. Além disto, a detecçäo dos auto-anticorpos anti-LDL- demonstra o potencial imunogênico desta partícula. Portanto, a detecçäo da LDL- e dos auto-anticorpos anti-LDL- abre novas perspectivas para o monitoramento dos fatores de risco para a aterosclerose vinculados às reaçöes oxidativas

Production and characterization of a monoclonal antibody against an Ascaris suum allergenic component

Ascaris suum allergenic components (PIII) separated by gel filtration chromatography of an adult worm extract were used to immunize BALB/c mice. Popliteal lymph node cells taken from the immunized animals were fused with SP2/O myeloma cells using polyethylene glycol (MW 1450) as fusogen. The hybridomas were cultured in HAT-containing medium and cloned at limiting dilutions. Supernatants from the growing hybrids were screened by ELISA using plates coated with PIII or the A. suum crude extract. The monoclonal antibody obtained, named MAC-3 (mouse anti-A. suum allergenic component), is an IgG1 kappa mouse immunoglobulin that specifically recognizes a 29,000 molecular weight protein (called allergenic protein) with an affinity constant of 1.7 x 10(9) M-1. The A. suum components recognized by MAC-3 induce specific IgE antibody production in immunized BALB/c mice. Ascitic fluid induced in Swiss mice by injecting ip the hybridoma cells and incomplete Freund's adjuvant was purified by affinity chromatography using a protein A-Sepharose column. The purified monoclonal antibody was then coupled to activated Sepharose beads in order to isolate the A. suum allergenic component from the whole extract by affinity chromatography

Antigenic cross-reactivity among components of Brazilian Elapidae snake venoms

Snake venoms from M. corallinus (LD5=7.1 + or - 0.83 µg), M.frontalis (LD50=19.3 + or - 3.13 µg), M. ibiboboca (LD50=19.8 + or - 2.07 µg) and M. spiixi (LD50=6.7 + or - 1.25 µg) (family Elapidae, genus Micrurus) injected into horses alone or in combination (M. corallinus with M. frontalis) elicit antibody production, as indicated in vivo by neutralization of venom lethality and in vitro by enzyme-linked immunosorbent assay (ELISA), immunoelectrophoresis (IE) and Western blotting (WB). Venom lethality was efficiently neutralized by the antisera, with the monovalent antivenoms being more efficient than the bivalent antivenom. Antibodies against venom components were detected by all artisera at different titers by ELISA. Upon IE, antisera against M. spiixi and M. frontalis venoms cross-reacted with the four types of venoms studied and recognized several molecular components, the precipitin lines obtained had distinct intensities and electrophoretic motilities, whereas the antivenom against M. corallinus only recognized components of its venom but not of the others. All antivenoms cross-reacted with all the elapid venoms in WB revealing several blands with distinct MWs in M. corallinus and M. spiixi venoms, two very sharp and separate bands in M. corallinus venom and a very sharp band of high MW together with several other smaller and faint bands in M. frontalis venom. The data indicate that snake venoms of the genus Micrurus are good immunogens that contain many cross-reactive molecules, and that their toxic components are neutralized more effectively by monovalent rather than by bivalent antivenom

A rapid and efficient purification method for horse IgG(T) usin a rat monoclonal antibody

1. Louvain rats (IgK-1a) were immunized with horse IgG(T). To generate mAb to IgG(T), popliteal lymph node cells taken from the immunized animals were fused to a non-secreting LOU/C immunocytoma (IR983F). The hybridomas were cultured in HAT -containing medium and cloned under limiting dilution conditions. Supernatants from the growing hybrids were screened by ELISE using plates coated with horse IgG(T) or IgGa+b+c. 2. The anti-IgG(T) mAb obtained was named LO-HoGT-1 (LOU anti-horse IgG(T)). It is an IgG2a rat antibody whose light chain allotype is IgK-1a, and with an affinity constant of 2.9 x 1010 M-1. 3. Ascites was isnduced in LOU (IgK-1b) rats by injecting the hybridoma cells and incomplete Freund's adjuvant ip. To obtain purified mAb, ascitic fluid was applied to a Sepharose anti-rat LOU IgK-1 a chain column. 4. The purified mAb was then coupled to Sepharose. Immunoelectrophoretically pure IgG(T) was obtained by passage of horse serum through this column. The entire procedure took less than 30 min and resulted in a highly purified IgG(T)

Isolation of horse IgG with protein A

Horse immunoglobulins were obtained from normal defatted with dextran sulfate and precipitated with ammonium sulfate. Eight mg of this preparation was submitted to affinity chromatography with protein A-Sepharose CL-4B. Low temperature (4-C) and a starting buffer at pH 8.0 were conditions required for all IgG subclasses to bind to protein A, even those with low affinity. The IgGs bound to protein A were eluted with glycine buffer at pH 2.8. The yield was about 90%. Its suggested that isolated IgG, instead of whole Igs, be used in serum therapy, reducing the amount of Igs and diminishing serum-related reactions

Teste psicomotor de continuacao de linhas. / Starting lines' psychomotor test

O teste de continuacao de linhas, da autoria do Dr. Jose Otavio de Freitas Jr., visa comprovar a organicidade de varios casos patologicos ate entao sem testes apropriados. O teste tem por objetivo a localizacao cerebral das lesoes nas areas frontais e pre-frontais atingidas. Baseada nos trabalhos de Luria, a pesquisa prosseguiu o estudo da aplicacao do teste em pessoas normais e com lesoes cerebrais