High-level expression of Actin of Paeonia lactiflora in Escherichia coli / 中草药

Objective: To construct the prokaryotic expression vector of Actin gene and to establish the high-level expression of Actin of Paeonia lactiflora in Escherichia coli. Methods: Based on cDNA sequence of Actin of P. lactiflora and the polyclonal sites on the prokaryotic expression vector pET-32a (+), a pair of PCR primers whose 5' end was inserted with BamH I and Hind III, respectively were designed. Subcloning of Actin was carried out using RT-PCR technique. The recombinant plasmid pMD18-T-PlActin and the prokaryotic expression vector pET-32a (+) was digested by BamH I and Hind III and the objective gene and the empty vector were purified. After ligation and transformation, the recombinant pET-32a (+)-PlActin, which was characterized by colony PCR, plasmid PCR, and double enzyme digestion, was transformed into BL21 (DE3), and the engineering expression strain was obtained. The expression of recombinant Actin protein in E. coli was induced at different concentration of inducer IPTG, different bacteria density, and different expression time. After SDS-PAGE and Coomassie brilliant blue R250 staining, the dry gel was prepared and the expression level of recombinant Actin protein was analyzed. Results: Subcloning sequencing result showed that the sequence of PlActin was exactly same with the objective sequence and the 5' and 3' ends were successfully inserted with BamH I and Hind III sites. The prokaryotic expression vector of Actin gene pET-32a (+)-PlActin was constructed successfully. The best concentration of IPTG was 0.1 mmol/L and the bacteria density A600 was 0.4 to 0.6. The optimal expression time was 4 h. Conclusion: The prokaryotic expression vector of Actin gene pET-32a (+)-PlActin is constructed and the high-level expression of Actin of P. lactiflora in E. coli is established.

Conservación de cultivos de hongos de importancia médica en agua destilada / Preservation of fungal cultures of medical importance in distilled water

Introducción: en la colección de cultivos de hongos patógenos del Instituto de Medicina Tropical Pedro Kourí, como centro de referencia nacional, se conservan los agentes causales de las principales micosis humanas, con fines de diagnóstico, investigaciones y enseñanza de la microbiología médica. La conservación de cultivos fúngicos en agua destilada estéril o método de Castellani ha sido avalada como un método que garantiza porcentajes elevados de viabilidad, pureza y estabilidad de las cepas; esto unido a su bajo costo y sencillez, la convierte en una alternativa ventajosa para el mantenimiento de los microorganismos en muchos laboratorios. Objetivo: mostrar los resultados en la preservación en agua destilada estéril de las principales especies fúngicas que integran esta colección. Métodos: se evaluó la viabilidad, pureza y estabilidad de las principales características morfológicas y fisiológicas de 240 cepas de diferentes especies fúngicas pertenecientes a la colección, conservadas en agua destilada estéril. Resultados: de los cultivos, 80 por ciento se conservó en estado viable y sin contaminaciones. Los mejores resultados se obtuvieron en la preservación de los hongos productores de abundantes conidios (97 por ciento de recuperación) y las levaduras (83 por ciento), mientras que con los dermatofitos y hongos dimórficos fue de 69 y 68 por ciento, respectivamente. En 17 géneros, la recuperación de cepas viables fue superior a 60 por ciento, mientras que en 8 resultó de 100 por ciento. El tiempo de conservación fue de 15 a 20 años. Se implementó una base de datos en formato digital de la colección sobre plataforma web. Conclusiones: el método de Castellani es un método de elección para la conservación de cultivos fúngicos en laboratorios de recursos limitados

Introduction: causal agents of the main human myicoses have been preserved in the culture collection of pathogenic fungi at Pedro Kourí Tropical Medicine Institute, a national reference center, with the purpose of using them for medical microbiology diagnoses, research and teaching. Preservation of fungal cultures in sterile distilled water, or Castellani's method, has been endorsed as a method ensuring high rates of strain viability, purity and stability, low costs and great simplicity. It is therefore an advantageous alternative for the maintenance of microorganisms in many laboratories. Objective: present the results obtained from the preservation in sterile distilled water of the main fungal species included in the collection. Methods: an evaluation was conducted of the viability, purity and stability of the main morphological and physiological characteristics of 240 strains of different fungal species from the collection, which had been preserved in sterile distilled water. Results: 80 percent of the cultures had retained their viable status without any contamination. The best results corresponded to the preservation of fungi producing abundant conidia (97 percent recovery) and yeasts (83 percent), followed by dermatophytes (69 percent) and dimorphic fungi (68 percent). In 8 genera, recovery of viable strains was 100 percent, and in 17 it was above 60 percent. Preservation time was 15-20 years. A web-based digital database was created for the collection. Conclusions: Castellani's is the method of choice for the preservation of fungal cultures in resource-limited laboratories

Bactéria ou parasita? a controvérsia sobre a etiologia da doença do sono e a participação portuguesa, 1898-1904 / Bacteria or parasite? the controversy over the etiology of sleeping sickness and the Portuguese participation, 1898-1904

A etiologia da doença do sono era desconhecida até o início do século XX. Essa doença tipicamente africana em breve se tornaria o principal obstáculo à colonização europeia. O envio de missões científicas às colônias para seu estudo in loco tornou-se inevitável. Portugal enviou a primeira missão de estudo, a Angola, em 1901, e a Royal Society of London apoiou duas missões britânicas de estudo da doença, em Entebe. O resultado dessas investigações estabeleceu uma controvérsia, na qual Portugal esteve envolvido de 1898 a 1904, no circuito nacional e internacional, objeto de análise deste artigo.

The etiology of sleeping sickness was unknown until the early twentieth century. This African disease soon became the main obstacle to European colonization. Sending scientific missions to the colonies to monitor its progression in loco thus became inevitable. Portugal sent the first research mission to Angola in 1901, and the Royal Society of London sponsored two British missions to study the disease in Entebbe (1902 and 1903). Their results led to a controversy in which Portugal was involved from 1898 to 1904, on the national and international circuits, analysed in this article.

Aldo Castellani y las expediciones de la Royal Society al país del sueño negro / Aldo Castellani and the expeditions of the Royal Society to the black sleep's country

At the beginning the investigation on infectious diseases was plenty of adventures in exotic countries. The efforts of the English investigators, headed by Patrick Manson, gave birth to the "tropical" medicine and "tropical" diseases, like the sleeping sickness, which was sweeping the country north to the Victoria Lake in 1901. The Royal Society of London sent two Commissions in search of the etiological agent. Aldo Castellani was decisive for the failure of the first - Low, Castellani, Christy,1902 - because even he saw Trypanosoma in samples of some patients, he did not appreciate his discovery; and decisive also for the success of the second -Bruce, Nabarro, Greig, 1903 - when he and Bruce recognized this Trypanosoma as the etiological agent. Following these expeditions, Low developed a brilliant career in England, Christy a life of investigation mixed up with adventures through Asia and Africa and Castellani a long life of lights and shadows in many lands.

En un comienzo la investigación sobre enfermedades infecciosas estuvo llena de aventuras en países exóticos. Impulsada por los investigadores ingleses, encabezados por Patrick Manson, nacieron la medicina y las enfermedades "tropicales", entre las cuales se encontraba la enfermedad del sueño, que a comienzos del siglo XX hacía estragos al norte del lago Victoria. La Real Sociedad de Londres envió dos Comisiones a Uganda para determinar el agente etiológico. Aldo Castellani fue decisivo para el fracaso de la primera, que incluía también a Low y Christy, en 1902, pues aunque vio tripanosomas en LCR de enfermos, no les otorgó valor y prefirió postular un diplococo como agente causal; y decisivo también para el éxito de la segunda, de Bruce, Nabarro y Greig, en 1903, al concordar con Bruce en que el tripanosoma era realmente el causante de la enfermedad. Después de estas expediciones, Low desarrolló una brillante carrera en Inglaterra, Christy una vida que combinaba investigación con aventura en Asia y África, y Castellani una larga vida de éxitos, oscurecida por sus ideas políticas, que lo ligaban a Mussolini.

A Case of Allergic Contact Dermatitis to Resorcin in Skin Marking Solution Occurred During Radiation Therapy / 대한피부과학회지

Resorcin is widely being used in the fields of the therapeutics, cosmetics, industry, but allergic contact dermatitis is an infrequent adverse reaction. We have experienced a case of allergic contact dermatitis to resorcin. A 59-year-old male with Kaposi's sarcoma on the right foot developed linear erythematous vesicular eruption along the marking areas 4 days after application of Castellanis paint, used as a skin marking solution for radiation therapy. He showed patch test positive reaction to Castellani's paint as well as its individual components, resorcin and phenol.