Plant metabolite 5-pentadecyl resorcinol is produced by the Amazonian fungus Penicillium sclerotiorum LM 5679 / O metabólito vegetal 5-pentadecil resorcinol é produzido pelo fungo amazônico Penicillium sclerotiorum LM 5679

Since the classic studies of Alexander Flemming, Penicillium strains have been known as a rich source of antimicrobial substances. Recent studies have identified novel metabolites produced by Penicillium sclerotiorum that have antibacterial, antifouling and pharmaceutical activities. Here, we report the isolation of a P. sclerotiorum (LM 5679) from Amazonian soil and carry out a culture-based study to determine whether it can produce any novel secondary metabolite(s) that are not thus-far reported for this genus. Using a submerged culture system, secondary metabolites were recovered by solvent extract followed by thin-layer chromatography, nuclear magnetic resonance, and mass spectroscopy. One novel secondary metabolite was isolated from P. sclerotiorum (LM 5679); the phenolic compound 5-pentadecyl resorcinol widely known as an antifungal, that is produced by diverse plant species. This metabolite was not reported previously in any Penicillium species and was only found once before in fungi (that time, in a Fusarium). Here, we discuss the known activities of 5-pentadecyl resorcinol in the context of its mode-of-action as a hydrophobic (chaotropicity-mediated) stressor.

Desde os estudos clássicos de Alexander Flemming, as cepas de Penicillium são conhecidas como uma fonte rica em substâncias antimicrobianas. Estudos recentes identificaram novos metabólitos produzidos pela espécie Penicillium sclerotiorum com atividades antibacteriana, anti-incrustante e farmacêutica. Aqui, relatamos o isolamento de uma colônia de P. sclerotiorum (LM 5679) do solo amazônico e relatamos também o estudo baseado em cultura para determinar se ele pode produzir qualquer novo metabólito (s) secundário (s) que não foram relatados até agora para este gênero. Usando um sistema de cultura submerso, os metabólitos secundários foram recuperados por extrato de solvente seguido por cromatografia em camada delgada, ressonância magnética nuclear e espectroscopia de massa. Um novo metabólito secundário foi isolado de P. sclerotiorum (LM 5679); o composto fenólico 5-pentadecil resorcinol que é amplamente conhecido como um antifúngico que é produzido por diversas espécies de plantas. Este metabólito não foi relatado anteriormente em nenhuma espécie de Penicillium, e foi encontrado apenas uma vez em fungos (Fusarium). Aqui, discutimos as atividades conhecidas do 5-pentadecil resorcinol no contexto de seu modo de ação como um estressor hidrofóbico (mediado pela caotropicidade).

Plant metabolite 5-pentadecyl resorcinol is produced by the Amazonian fungus Penicillium sclerotiorum LM 5679

Abstract Since the classic studies of Alexander Flemming, Penicillium strains have been known as a rich source of antimicrobial substances. Recent studies have identified novel metabolites produced by Penicillium sclerotiorum that have antibacterial, antifouling and pharmaceutical activities. Here, we report the isolation of a P. sclerotiorum (LM 5679) from Amazonian soil and carry out a culture-based study to determine whether it can produce any novel secondary metabolite(s) that are not thus-far reported for this genus. Using a submerged culture system, secondary metabolites were recovered by solvent extract followed by thin-layer chromatography, nuclear magnetic resonance, and mass spectroscopy. One novel secondary metabolite was isolated from P. sclerotiorum (LM 5679); the phenolic compound 5-pentadecyl resorcinol widely known as an antifungal, that is produced by diverse plant species. This metabolite was not reported previously in any Penicillium species and was only found once before in fungi (that time, in a Fusarium). Here, we discuss the known activities of 5-pentadecyl resorcinol in the context of its mode-of-action as a hydrophobic (chaotropicity-mediated) stressor.

Resumo Desde os estudos clássicos de Alexander Flemming, as cepas de Penicillium são conhecidas como uma fonte rica em substâncias antimicrobianas. Estudos recentes identificaram novos metabólitos produzidos pela espécie Penicillium sclerotiorum com atividades antibacteriana, anti-incrustante e farmacêutica. Aqui, relatamos o isolamento de uma colônia de P. sclerotiorum (LM 5679) do solo amazônico e relatamos também o estudo baseado em cultura para determinar se ele pode produzir qualquer novo metabólito (s) secundário (s) que não foram relatados até agora para este gênero. Usando um sistema de cultura submerso, os metabólitos secundários foram recuperados por extrato de solvente seguido por cromatografia em camada delgada, ressonância magnética nuclear e espectroscopia de massa. Um novo metabólito secundário foi isolado de P. sclerotiorum (LM 5679); o composto fenólico 5-pentadecil resorcinol que é amplamente conhecido como um antifúngico que é produzido por diversas espécies de plantas. Este metabólito não foi relatado anteriormente em nenhuma espécie de Penicillium, e foi encontrado apenas uma vez em fungos (Fusarium). Aqui, discutimos as atividades conhecidas do 5-pentadecil resorcinol no contexto de seu modo de ação como um estressor hidrofóbico (mediado pela caotropicidade).

Plant metabolite 5-pentadecyl resorcinol is produced by the Amazonian fungus Penicillium sclerotiorum LM 5679 / O metabólito vegetal 5-pentadecil resorcinol é produzido pelo fungo amazônico Penicillium sclerotiorum LM 5679

Since the classic studies of Alexander Flemming, Penicillium strains have been known as a rich source of antimicrobial substances. Recent studies have identified novel metabolites produced by Penicillium sclerotiorum that have antibacterial, antifouling and pharmaceutical activities. Here, we report the isolation of a P. sclerotiorum (LM 5679) from Amazonian soil and carry out a culture-based study to determine whether it can produce any novel secondary metabolite(s) that are not thus-far reported for this genus. Using a submerged culture system, secondary metabolites were recovered by solvent extract followed by thin-layer chromatography, nuclear magnetic resonance, and mass spectroscopy. One novel secondary metabolite was isolated from P. sclerotiorum (LM 5679); the phenolic compound 5-pentadecyl resorcinol widely known as an antifungal, that is produced by diverse plant species. This metabolite was not reported previously in any Penicillium species and was only found once before in fungi (that time, in a Fusarium). Here, we discuss the known activities of 5-pentadecyl resorcinol in the context of its mode-of-action as a hydrophobic (chaotropicity-mediated) stressor.

Desde os estudos clássicos de Alexander Flemming, as cepas de Penicillium são conhecidas como uma fonte rica em substâncias antimicrobianas. Estudos recentes identificaram novos metabólitos produzidos pela espécie Penicillium sclerotiorum com atividades antibacteriana, anti-incrustante e farmacêutica. Aqui, relatamos o isolamento de uma colônia de P. sclerotiorum (LM 5679) do solo amazônico e relatamos também o estudo baseado em cultura para determinar se ele pode produzir qualquer novo metabólito (s) secundário (s) que não foram relatados até agora para este gênero. Usando um sistema de cultura submerso, os metabólitos secundários foram recuperados por extrato de solvente seguido por cromatografia em camada delgada, ressonância magnética nuclear e espectroscopia de massa. Um novo metabólito secundário foi isolado de P. sclerotiorum (LM 5679); o composto fenólico 5-pentadecil resorcinol que é amplamente conhecido como um antifúngico que é produzido por diversas espécies de plantas. Este metabólito não foi relatado anteriormente em nenhuma espécie de Penicillium, e foi encontrado apenas uma vez em fungos (Fusarium). Aqui, discutimos as atividades conhecidas do 5-pentadecil resorcinol no contexto de seu modo de ação como um estressor hidrofóbico (mediado pela caotropicidade).

Nó-de-cachorro (Heteropterys tomentosa A. Juss.): espécie de uso medicinal em Mato Grosso, Brasil / Nó-de-cachorro (Heteropterys tomentosa A. Juss.): a medicinal plant from Mato Grosso State, Brazil

Neste estudo é apresentada uma revisão sobre nó-de-cachorro (Heteropterys tomentosa A. Juss.), espécie de uso medicinal em Mato Grosso. São abordadas as características da espécie com a descrição da planta, hábitat, distribuição geográfica, ecologia, fenologia e biologia reprodutiva, propriedades químicas e farmacológicas. Os aspectos como a etnobotânica (uso e partes usadas), formas de exploração atual e informações sobre cultivo são discutidos.

This study presents a review about "Nó-de-cachorro" (Heteropterys tomentosa A. Juss.), a medicinal species from Mato Grosso State, Brazil. We outlined the characteristics of the species including the plant description, habitat, geographic distribution, ecology, phenology and reproductive biology, chemical and pharmacological properties. Aspects such as ethnobotany (use and used parts), current ways of exploiting and information on cultivation are discussed.

Estudo in vitro da atividade do extrato etanólico de sementes de bacupari (Rheedia gardneriana Planch. & Triana) e das frações no crescimento de Streptococcus mutans / In vitro study of the activity of ethanol extract from bacupari (Rheedia gardneriana Planch. & Triana) seeds and its fractions on Streptococcus mutans growth

Streptococcus mutans, principal microrganismo da cavidade oral, desempenha papel preponderante na formação de placas dentárias, sendo considerado o agente etiológico primário da cárie. Rheedia gardneriana, conhecida popularmente como bacupari, é uma planta utilizada com fins medicinais para o tratamento de diversas patologias, e por apresentar atividade antimicrobiana de compostos das folhas contra bactérias Gram-positivas e Gram-negativas. O objetivo do presente trabalho foi avaliar o efeito de extrato de semente de R. gardneriana sobre a cepa S. mutans UA159. Os testes foram conduzidos com o extrato etanólico bruto e as frações obtidas com os solventes diclorometano, etanol-água, metanol e hexano, em ensaios de inibição in vitro. O extrato bruto (100 por cento) apresentou halos de inibição com diâmetro similar ao obtido com solução de digluconato de clorexidina 0,12 por cento, usada como controle. Os ensaios com a fração diclorometano exibiram atividade inibitória 35 por cento menor comparado com o controle, enquanto nenhum efeito antimicrobiano foi observado com a fração etanol-água. Contrariamente, os resultados obtidos com as frações hexânica e metanólica demonstraram claramente a atividade antimicrobiana por inibição do crescimento bacteriano. Na fração metanólica a formação de halos de inibição foi similar ao do controle. Estes dados apresentam atividade antimicrobiana de R. gardneriana contra S. mutans.

Streptococcus mutans, which is the main microorganism of the oral cavity, plays a preponderant role in dental plaque formation and is considered the primary etiologic agent regarding caries. Commonly known as "bacupari", Rheedia gardneriana is a plant used for medicinal purposes in the treatment of several pathologies; besides, its leaves have compounds that present antimicrobial activity against Gram-positive and Gram-negative bacteria. The aim of this work was to evaluate the effect of R. gardneriana seed extract on S. mutans strain UA159. The tests were carried out with crude ethanol extract and the fractions obtained with the solvents dichloromethane, ethanol-water, methanol, and hexane in in vitro inhibition assays. The crude extract (100 percent) presented inhibition halos with diameter similar to that obtained by using 0.12 percent chlorhexidine digluconate solution as control. Assays with the fraction dichloromethane showed an inhibitory activity 35 percent lower than that of the control, whereas no antimicrobial effect was observed with the ethanol-water fraction. Conversely, the results obtained with the fractions hexane and methanol clearly demonstrated antimicrobial activity by inhibiting the bacterial growth. In the methanol fraction, the formation of inhibition halos was similar to that in the control. These data present antimicrobial activity of R. gardneriana against S. mutans.

Results of CHOP chemotherapy for diffuse large B-cell lymphoma

Patients with diffuse large B-cell lymphoma treated in a University Hospital were studied from 1990 to 2001. Two treatment regimens were used: ProMACE-CytaBOM and then, from November 1996 on, the CHOP regimen. Complete remission (CR), disease-free survival (DFS), and overall survival (OS) rates were determined. Primary refractory patients and relapsed patients were also assessed. A total of 111 patients under 60 years of age were assessed and ranked according to the international prognostic index adjusted to age. Twenty (18 percent) of them were classified as low risk, 40 (36 percent) as intermediate risk, 33 (29.7 percent) as high intermediate risk, and 18 (16.3 percent) as high risk. Over a five-year period, OS and DFS rates were 71 and 59 percent, respectively, for all patients. For the same time period, OS and DFS rates were 72.8 and 61.3 percent, respectively, for 77 patients treated with CHOP chemotherapy and 71.3 and 60 percent for patients treated with the ProMACE-CytaBOM protocol. There was no significant difference in OS or DFS between the two groups. Eleven of 50 refractory and relapsed patients were consolidated with high doses of chemotherapy. Three received allogenic and 8 autologous bone marrow transplantation. For the latter, CR was 62.5 percent and mean OS was 41.1 months. The clinical behavior, CR, DFS, and OS of the present patients were similar to those reported in the literature. We conclude that both the CHOP and ProMACE-CytaBOM protocols can be used to treat diffuse large B-cell lymphoma patients, although the CHOP protocol is preferable because of its lower cost and lower toxicity.

Plantas hipoglicemiantes utilizadas por comunidades tradicionais na Bacia do Alto Paraguai e Vale do Guaporé, Mato Grosso-Brasil

No Brasil, com inúmeras espécies tropicais, evidencia-se o conhecimento e uso da vegetação medicinal pelas comunidades quilombolas, ribeirinhas, rurais, tradicionais e indígenas, que são praticadas até os dias atuais. O Estado de Mato Grosso pela sua localização no Planalto Central brasileiro possui uma variedade destas comunidades, que utilizam estas plantas medicinais, levando em consideração o conhecimento popular passado de geração em geração. Esta pesquisa visa identificar a forma de coleta e os usos desse grupo vegetal indicados pelos usuários. O estudo foi realizado em 15 comunidades tradicionais (não indígenas) na Bacia do Alto Paraguai e 2 (duas) no Vale do Guaporé. A coleta de dados deu-se através de líderes comunitários, benzedeiras, parteiras e usuários utilizando o método qualitativo, mediante abordagem qualitativa, com auxílio de entrevistas estruturadas, semi-estruturadas e abertas. Dentre as espécies identificadas 17 estacam-se para o tratamento do Diabetes, dentre elas citam-se: Anacadium humile; Bauhinia glabra; Cecropia pachystachya; Hancornia speciosa; Heteropteris aphrodisiaca; Leonotis nepetifolia; Momordica charantia; Solanum lycocarpum. Registra-se ainda que as partes mais utilizadas desses vegetais são: folhas, casca do caule, raiz, planta toda, brotos, óleos dos frutos e polvilho dos frutos. De acordo com os usuários a maioria das plantas não podem ser colhidas após o nascer do sol e o preparo deve ser feito com folhas secas ou que tenham sido submetidas ao processo de secagem sobre o fogão de lenha ou em local abafado.

In Brazil, with a large number of tropical species, evidenciate the knowledge and use of the medicine vegetation by the "quilombolas", riverine, rurally, traditional and indigenous communities are used nowadays. The Mato Grosso state by their localization in the Central Brazilian Plateau posses a variety of community, which use those medicinal plants, carrying in consideration the popular knowledge passed though generation to generation. This research had as purpose to identificate the use of this vegetable group the collect ways related by the users and collectors. The study was realized into 15 traditional communities (not indigenous) in the Alto Paraguay Bay and 2 (two) in the Guaporé Valley. The data collect were made with the community leaders, women healers, midwife and users, it was used the qualitative method, with qualitative boarding, with aim of structured qualitative interview, semistructured and opened. Between the identified 17 destacated to the treatment of diabetes, between them it was cited Anacadium humile; Bauhinia glabra; Cecropia pachystachya; Hancornia speciosa; Heteropteris aphrodisiaca; Leonotis nepetifolia; Momordica charantia; Solanum lycocarpum. It was registered still, that the more used parts of this vegetable are: leaves, burke, root, all plant, shoot, fruit oil and fruits flour. In according to users the biggest part of the plants can't be collected after the sun rinsing and the prepare must be done with dry leaves or that had been submitted to the process of drying on the oven or in a muffled place.

Plantas medicinais usadas para tratamentos dermatológicos, em comunidades da Bacia do Alto Paraguai, Mato Grosso

O uso de plantas medicinais no Brasil e em especial no Estado de Mato Grosso vem sendo registrado em suas comunidades rurais e tradicionais. Este trabalho visa identificar plantas com potencial dermatológico utilizada pelas comunidades: quilombolas, ribeirinhas, rurais e tradicionais dos municípios de Barão de Melgaço, Nossa Senhora do Livramento, Poconé e Santo Antônio de Leverger. Fez-se a coleta de material botânico fértil e levantamento sobre o uso desses vegetais com a população local. Este material encontra-se identificado e depositado em Coleção de Referência de Plantas Medicinais no UFMT, Herbário Central. Registraram-se 36 espécies utilizadas no tratamento de doenças como afecções, feridas e úlceras, erisipelas, herpes labial e verrugas. Destacam-se espécies como: Astronium fraxinifolium Schott.; Bixa orellana L.; Brosimum gaudichaudii Tréc.; Cassia grandis Lf.; Heliotropium indicum L.; Jacaranda brasiliana (Lam.) Pers.; Macrosiphonia velame (St. Hil.) Mull. Arg.; Pistia stratiotes L.; Senna alatal. Roxb., entre outras. Estes vegetais são freqüentemente usados na forma de banhos e chás. O órgão vegetal mais utilizado foi as folhas, seguido de cascas do caule, frutos, óleo, raiz, planta inteira, seiva e sementes. O avanço da fronteira agropecuária nesse Estado é constante preocupação dos botânicos e das comunidades que ao longo das décadas usufruem desse potencial medicinal.

Medicinal plants used for dermatology treatment in communities of Alto Rio Paraguai' Basin in Mato Grosso State. The use of medicinal plants in Brazil and in special in the Mato Grosso State is being registered by the rural and traditional communities. This work aims to identify plants with dermatologic potential used by the communities: quilombolas, riversides, rural and traditional of the municipal districts of Barão de Melgaço, Nossa Senhora do Livramento, Poconé and Santo Antonio de Leverger. The collection of fertile botanical material and rising on the use of these vegetables was done with the local population. The identified material is deposited in the Reference Collection of Medicinal Plants in UFMT Central Herbarium. It were registered 36 species used in the diseases treatment as infections, hurt and ulcers, erysipelas, labial herpes and warts. Highlight species as: Astronium fraxinifolium Schott.; Bixa orellana L.; Brosimum gaudichaudii Tréc.; Cassia grandis Lf.; Heliotropium indicum L.; Jacaranda brasiliana (Lam.) Pers.; Macrosiphonia velame (St. Hil.) Mull. Arg.; Pistia stratiotes L.; Senna alata (l.) Roxb., come in another. These vegetables are frequently used in baths and teas form. The used vegetable organ was to the leaves followed by the peel and of the stem, fruits, oil, root, the whole plant, sap and seeds. The advance of the agriculture in this state is an constant preoccupation of the botanists and communities who along decades use those medicinal potential.

Extracts of Ascaris suum egg and adult worm share similar immunosuppressive properties

Adult Ascaris suum body extract (Asc) prepared from male and female worms (with stored eggs) down-regulates the specific immune response of DBA/2 mice to ovalbumin (OA) and preferentially stimulates a Th2 response to its own components, which is responsible for the suppression of the OA-specific Th1 response. Here, we investigated the participation of soluble extracts prepared from male or female worms or from eggs (E-Asc) in these immunological events. Extracts from either sex (1 mg/animal) or E-Asc (0.35 or 1 mg protein/animal) suppressed the delayed-type hypersensitivity (DTH) reaction (60-85 percent), proliferative response (50-70 percent), IL-2 and IFN-gamma secretion (below detection threshold) and IgG1 antibody production (70-90 percent) of DBA/2 mice to OA. A dose of 0.1 mg E-Asc/animal did not change DTH or proliferation, but was as effective as 0.35 mg in suppressing IL-2 and IFN-gamma, and OA-specific IgG1 antibodies. Lymph node cells from DBA/2 mice injected with Asc (1 mg/animal) or a high dose of E-Asc (1 mg protein/animal) secreted IL-4 upon in vitro stimulation with concanavalin A. As previously demonstrated for Asc, the cytokine profile obtained with the E-Asc was dose dependent and changed towards Th1 when a low dose (0.1 mg protein/animal) was used. Taken together, these results suggest that adult worms of either sex and eggs induce the same type of T cell response and share similar immunosuppressive properties

Corn oil favours protease production by Streptomyces viridosporus T7A

Besides of being largely used for antibiotic production, streptomyces have also been pointed out as good producers of enzymes with industrial interest such as protease. In this work, the effect of corn oil on protease production by S. virisdoporus T7A was investigated as part of a wide project for microbial protease production. Culture media contained 0.65(per cent) yeast as nitrogen source, corn oil or corn oil combined with 0.65(per cent) glucose as carbone source, plus mineral salts. In both cases, corn oil used in three differents concentrations, 0.1, 0.5 and 1(per cent) (p/V). All experiments were carried out in agitated flasks at 37§C for 105 hours. Higher protease activty (52 U/L) was obtained in medium containing 0.65(per cent) glucose and 1.0(per cent) corn oil as carbon sources. Protease activity responded positively to the increase in the medium C/N ratio, i.e., to the increment in oil concentration. Our results also suggested that corn oil favours enzyme stability during the fermentation.

Traumatismo cerrado de tórax: Tratamiento quirúrgico de la ruptura bronquial de diagnóstico tardío / Non penetrating thoracic trauma: Delayed diagnosis and treatment of bronchial rupture

La ruptura bronquial secundaria a traumatismo cerrado de tórax es poco frecuente y muchas veces causa la muerte del paciente.Es menos frecuente aún su diagnóstico tardío,que haya pasado desapercibido inicialmente.Se presentan dos casos de diagnóstico tardío(entre 2 meses y 2 años de evolución de su traumatismo)con atelectasia pulmonar cuyos estudios determinaron la sección completa del bronquio fuente.El tratamiento quirúrgico resultó favorable,con buena expansión pulmonar a largo plazo.

Criopreservaçäo de medula óssea e células pluripotentes periféricas utilizando um congelador programável: experiência em 86 congelamentos / Cryopreservation of bone marrow and peripheral blood stem cells using a controlled rate freezing system: experience on 86 procedures

A infusao de células hematopoéticas totipotentes criopreservadas permite a recuperaçao da hematopoese após quimioterapia mieolblativa. Objetivo. A formaçao de cristais de gelo durante o processo de congelamento é o fator principal que causa ruptura das estruturas celulares. A criopreservaçao dessas células a uma taxa constante preveniria os danos causados pelo congelamento brusco. Métodos. Vinte e três pacientes com mediana de 25 anos (variaçao 3-57) tiveram a medula óssea e/ou células-tronco periféricas (CTP) coletadas no período de março de 1993 a outubro de 1994, totalizando 86 congelamentos. Os pacientes apresentavam as seguintes neoplasias: linfoma nao-Hodgkin (n=5), leucemia mielóide aguda (n=8), leucemia linfóide aguda (n=6), doença de Hodkin (n=3) e mieloma múltiplo (n=1). O congelamento foicontrolado por um computador, acoplado ao sistema, às seguintes temperaturas: -1 graus Celsius/min até -45 graus Celsius e depois a -10 graus Celsius/min até -80 graus Celsius. Após o congelamento, as células foram mantidas em freezer a -110 graus Celsius até o momento da infusao. Para obtençao das CTP, empregou-se o fator de crescimento estimulante de granulócitos (G-CSF). Resultados. Uma mediana de 3,16 x 10(8) céls./kg (variaçao 0,86-24,22) de CTP e 2,03 x 10(8) céls./kg (variaçao 0,19-12,21) de medula óssea foi congelada. A mediana para atingir granulócitos maior ou igual a 500/muL e plaquetas maior que 20.000/muL foi de 12 dias (variaçao 8-40) e 31 dias (variaçao 8-80), respectivamente. Todos os pacientes tiveram recuperaçao hematopoética após a infusao das células criopreservadas. Conclusao. A criopreservaçao em congelador program vel permite o armazenamento de células hematopoéticas e, potencialmente, pode causar menor dano celular.

Purification and properties of storage proteins (vicilins) from cowpea (Vigna unguiculata) seeds which are susceptible or resistant to the bruchid beethe Callosobruchus maculatus

Vicilins (7S storage proteins) from seeds of cowpea (Vigna unguiculata) cultivars which are susceptible or resistant to the bruchil veetle C. maculatus were purified by size-exclusion and ion-exchange chromatography. The vicilins were partially characterized by polyacrylamide gel electrophoresis under both denaturing and non-denaturing conditions, by Western blotting and by amino acid analysis. The variant vicilins from C. maculatus-resistant seeds do not differ appreciably from vicilins from susceptible seeds by these criteria except that they are more strongly bound to DEAE-Sepharose, suggesting differences in charge between the various molecules

Colorimetric assay for the measurement of thymocyte/thymic stromal cell adhesion

1. We describe a simple and accurate colorimetric adhesion assay (CAA) and illustrate the assay by measuring the adhesion of mouse thymocytes to mouse 2BH4 cells. 2. The assay is based on the crystal violet staining of thymocytes adhered to a subconfluent layer of 2BH4 cells (plated at 2 x 10(4) cells/well for 24 h). The optimal incubation time was shown to be 1 h and washing in PBS of non-bound and non-specifically bound thymocytes is the critical step for the precision and accuracy of the assay. 3. Saturation curves were obtained for thymocytes adhered to plated 2BH4 cells. The blank (only 2BH4 cells) was near 0.200 +/- 0.010 (mean +/- SD) and was quite reproducible. As expected, the extent of adhesion was also dependent on the number of plated 2BH4 cells. Standard curves need to be run with each assay for quantitative measurements. The intra-assay and interassay coefficients of variation were 5 and 20, respectively. 4. The specificity of the reaction was demonstrated by the reduction of adherence by trypsin pretreatment of thymocytes, and the dose-dependent inhibition of adherence by rabbit anti-mouse thymocyte antisera but not rabbit anti-mouse immunoglobulin antisera. 5. The proposed method is simple and requires less effort than the counting of adhered cells with the light microscope and does not require the use of radioactive material as when labelling with Na2(51)CrO4 is utilized.

Suppression of the IgE antibody response by Ascaris suum components: effect of x-irradiation

The effect of X-irradiation on the supperession of IgE antibody responses induced by some of the Ascaris suum (ASC) components was analyzed in mice (7-week old A?Sn females). Treatment with 300 R 24h before immunization with 50 *g OVA and 200 *g ASC suppressive components abolished the damping effect on ati-OVA IgE antibody levels. The same effect was observed on the anti-ASC IgE antibody response obtained in mice injected with 200 *g ASC immunogenic plus 200 *g ASC suppressive components. Moreover, the failure of suppressive components to induce an IGE anti-ASC antibody response on their own was also abolished by X-irradiation. These results indicate that the suppressive components are able to elicit an IgE antibody response, but simultaneously activate a regulatory mechanism which suppressive both the homologous (anti-ASC) and heterologous (anti-OVA) antibody formation

Paf antagonists do not modify IgE antibody production in mice

The effect of selective PAF antagonists on the in vivo production of IgE antibodies was investigated. The anti-ovalbumin IgE antibody content was estimated by passive cutaneous anaphylactic reaction (PCA) in the plasma of Balb/c mice 10 days after immunization with ovalbumin and alum. The PAF antagonists, BN 52021 (5 mg/kg, ip), BN 50730(20 mg/kg, ip) BN 50730 (20 mg/kg, po), WEB 2086 (2 mg/kg, ip) and WEB 2170 (5 mg/kg, ip) were administered 1 h before immunization and twice a day for 8 days thereafter. The effect of the antagonists on the PAF-induced vasopermeability was also assayed. In the immunized mice the level of antiovalbumin IgE antibody, estimated by PCA titer, was 1/640. The treatment with the PAF antagonists did not change this level. At the concentrations employed, the antagonists BN 50730, WEB 2086 and WEB 2170 significantly reduced the PAF-induced vascular permeability. These results suggest that PAF does not seem to have a relevant effect on the production of IgE antibodies in vivo in the system used in the present study

Further characterization of ascaris suum component(s) with supressive activity on the IgE antibody response

In order to characterize the component(s) of Ascaris suum reponsible for damping of the IgE antibody production we demonstrated that the extract incubated in sodium acetate buffer, pH 4.5, maintained its suppressive effect and the same protein banding pattern by SDS-PAGE. Elimination of the lipoprotein components of the extract also left its damping properties unchanged. SDS-PAGE of the lipoprotein-free extract revealed practically the same pattern as shown by the whole extract, except for the high molecular weight polupeptides. These results indicate that the suppressive component(s) of A. suum did not precipitate and retained their activity at low pH. In addition, they appear not to be lipoproteins

Murine delayed type hypersensitivity is suppressed by Ascaris suum extract

We studied the suppressive effect of an Ascaris suum extract on delayed-type hypersensitivity (DTH) reactions to ovalbumin (OVA) and to Bacillus Calmette-Guerin (BCG). The ability of mice to develop DTH reactions to both antigens was suppressed when an immunizing dose of the antigen was given subcutaneously together with the Ascaris extract. Partial or complete suppression of the response to OVA was obtained by the use of 400 por 1000 microng of Ascaris extract, respectively. The response to BCG, on the other hand, was totally suppressed by 400 microng of extract

Suppression of IgE antibody production by Ascaris suum extract: characterization of suppressive component(S)

Partial characterization ot the suppressive component(s) of A. suum extract that is (are) responsible for damping production of IgE antibody to ovalbumin was performed by physical and chemical methods. Digestion of the whole extract with trypsin and chymotrypsin completely abolished the suppressive activity. Oxidation with sodium metaperiodate or heating at 56-C, however, had no effect. These results indicate that the integrity of heat-stable proteins(s) present in the crude extract is essential for its suppressive effect. In addition, the carbohydrate moiety does not seem to play an important role in this effect