Escherichia coli expression and purification of four antimicrobial peptides fused to a family 3 carbohydrate-binding module (CBM) from Clostridium thermocellum

Antimicrobial peptides (AMPs) are molecules that act in a wide range of physiological defensive mechanisms developed to counteract bacteria, fungi, parasites and viruses. Several hundreds of AMPs have been identified and characterized. These molecules are presently gaining increasing importance, as a consequence of their remarkable resistance to microorganism adaptation. Carbohydrate-binding modules (CBMs) are non-catalytic domains that anchor glycoside hydrolases into complex carbohydrates. Clostridium thermocellum produces a multi-enzyme complex of cellulases and hemicellulases, termed the cellulosome, which is organized by the scaffoldin protein CipA. Binding of the cellulosome to the plant cell wall results from the action of CipA family 3 CBM (CBM3), which presents a high affinity for crystalline cellulose. Here CipA family 3 CBM was fused to four different AMPs using recombinant DNA technology and the fusion recombinant proteins were expressed at high levels in Escherichia coli cells. CBM3 does not present antibacterial activity and does not bind to the bacterial surface. However, the four recombinant proteins retained the ability to bind cellulose, suggesting that CBM3 is a good candidate polypeptide to direct the binding of AMPs into cellulosic supports. A comprehensive characterization of the antimicrobial activity of the recombinant fusion proteins is currently under evaluation.Instituto de Tecnologia Química e Biológica,Fundação para a Ciência e a Tecnologia (FCT

Translational control of UIS4 protein of the host-parasite interface is mediated by the RNA binding protein Puf2 in Plasmodium berghei sporozoites

Copyright: © 2016 Silva et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.UIS4 is a key protein component of the host-parasite interface in the liver stage of the rodent malaria parasite Plasmodium berghei and required for parasite survival after invasion. In the infectious sporozoite, UIS4 protein has variably been shown to be translated but also been reported to be translationally repressed. Here we show that uis4 mRNA translation is regulated by the P. berghei RNA binding protein Pumilio-2 (PbPuf2 or Puf2 from here on forward) in infectious salivary gland sporozoites in the mosquito vector. Using RNA immunoprecipitation we show that uis4 mRNA is bound by Puf2 in salivary gland sporozoites. In the absence of Puf2, uis4 mRNA translation is de-regulated and UIS4 protein expression upregulated in salivary gland sporozoites. Here, using RNA immunoprecipitation, we reveal the first Puf2-regulated mRNA in this parasite.This work was supported by Fundação para a Ciência e a Tecnologia (FCT) grants PTDC/SAU-MIC/122082/2010 and PTDC/BIA-BCM/105610/2008 to GRM, and SFRH/BPD/72619/2010 to PAGCS.info:eu-repo/semantics/publishedVersio

Today, in the endoscopist hands

Endoscopic submucosal dissection (ESD) was first described as a non-surgical promise for early gastric epithelial lesions

Handling strategies for termites in the Azores

As térmitas estão perfeitamente estabelecidas nos Açores e constituem já uma praga de dimensões apreciadas nas zonas urbanas. Com base no conhecimento actual, é impossível erradicar as térmitas dos Açores. São conhecidas, actualmente, três espécies de térmitas nos Açores: uma térmita de madeira seca, a Cryptotermes brevis, uma térmita de madeira viva, a Kalotermes flavicollis, e uma térmita subterrânea, a Reticulitermes grassei. Qualquer que seja a estratégia de infestação, todas estas espécies irão causar grandes prejuízos económicos aos açorianos, nas próximas décadas, sendo as formas de combate e gestão diferentes para as três espécies. Com base na investigação científica realizada no último ano, em relação à espécie de térmita de madeira seca, Cryptotermes brevis, torna-se claro que as principais estratégias de gestão passam por: 1) evitar a dispersão das espécies entre ilhas; 2) melhorar as medidas de quarentena para, assim, evitar a entrada de novas espécies no arquipélago; 3) melhorar as técnicas de inspecção e treinar novos inspectores; 4) educar as pessoas para lidarem com a gestão dos insectos alados; 5) tratar o mobiliário com uma série de técnicas recentemente disponibilizadas (Bolhas de gás com CO2 ou N2) ou através do simples uso do frio, bem como do calor do sol; 6) remover as peças de madeira muito infestadas e proceder à sua substituição por peças de metal ou madeira pré-tratada com químicos em autoclave; 7) usar químicos em tratamentos localizados, apenas, quando a infestação tiver um carácter de fraca ou média infestação e ter sempre o cuidado de seleccionar o químico considerado mais adequado com base nos dados experimentais; 8) testar a possibilidade de utilizar medidas de controlo em grande escala, com base no uso de fumigação por gases ou por calor; 9) criar nova legislação.ABSTRACT: Termites are well-established and are serious pest of structural wood in the Azores. Eradication of termites cannot currently be achieved. Three species of termites are currently known from the archipelago (dry-wood termite Cryptotermes brevis, living trees Kalotermes flavicollis and subterranean termite Reticulitermes grassei). Whatever the infestation strategy, all these three species will cause enormous economic impact in the next decades in the Azores, and the types of control that are needed are entirely different for the three. As a result of the scientific research performed with the dry-wood termite Cryptotermes brevis in the last year it is now clear that the main management strategies are: 1) to avoid further dispersal between islands; 2) to improve quarantine measures in order to avoid the entrance in the archipelago of new termite species; 3) to improve inspection techniques and train new inspectors; 4) to educate home-owners to manage alates spread; 5) to treat furniture with newly available techniques (e.g., Gas Bubble with non-toxic physical controls such as CO2, or N2 gas) or simple use of "sun heat" or extreme cold; 6) to remove all highly infested wood and replace it with new materials such as metal parts or pressure-treated wood when (re)building; 7) to use chemicals in spot-treatments only when the infestation is moderate to small and selecting the most adequate chemical based on experimental grounds; 8) to test the possibility of using in the Azores whole-block control usually by gas or heat fumigation; 9) to create new legislation

Síndrome de transfusão feto-fetal: neurodesenvolvimento de lactentes tratados com cirurgia a laser

To assess the neurodevelopmental functions of survivors of twin-twin transfusion syndrome (TTTS) treated by fetoscopic laser coagulation (FLC), during the first year of life, comparing them to a control group; and to verify the influence of specific variables on neurodevelopment. Method This was a prospective, longitudinal study. The sample comprised 33 monochorionic diamniotic twins who underwent FLC for treatment of TTTS and 22 full-term infants of single-fetus pregnancies. Bayley Scales of Infant and Toddler Development Screening Test were used for evaluation. Prenatal, perinatal and postnatal information were obtained. Results There was an increased frequency of infants in the TTTS group with inadequate performance compared to the control group. The identified variables (fetal donor, low economic income and cardiorespiratory disease) negatively impacted expressive communication and fine motor skills. Conclusion Although through follow-up is recommended in all TTTS survivors, particular attention is required for the high-risk group as defined in this study744307313Avaliar o desenvolvimento neurológico de sobreviventes da sindrome de transfusão feto-fetal (STFF) submetidos à coagulação a laser por fetoscopia (CLF), durante o primeiro ano de vida, comparando estes ao grupo controle; e verificar a influência de variáveis específicas no desenvolvimento. Método Tratou-se de estudo prospectivo, longitudinal. A amostra foi composta por 33 gêmeos diamnióticos monocoriônicos submetidos à CLF para tratamento da STFF e 22 lactentes a termo de gestação única. Bayley Scales of Infant and Toddler Development Screening Test foram utilizadas para avaliação. Informações pré-natal, perinatal e pós-natal foram coletadas. Resultados Houve maior número de lactentes com desempenho inadequado no grupo STFF do que no controle. As variáveis identificadas (feto doador, baixa renda econômica e doença cárdio-respiratória) influenciaram negativamente a comunicação expressiva e as habilidades motoras finas. Conclusão Embora o acompanhamento seja recomendado para todos lactentes com STFF, especial atenção deve ser dada àqueles que apresentam fatores de risc