Molecular characterization of a crustinlike antimicrobial peptide in the giant tiger shrimp, Penaeus monodon, and its expression profile in response to various immunostimulants and challenge with WSSV

A crustinlike antimicrobial peptide from the haemocytes of giant tiger shrimp, Penaeus monodon was partially characterized at the molecular level and phylogenetic analysis was performed. The partial coding sequence of 299 bp and 91 deduced amino acid residues possessed conserved cysteine residues characteristic of the shrimp crustins. Phylogenetic tree and sequence comparison clearly confirmed divergence of this crustinlike AMP from other shrimp crustins. The differential expression of the crustinlike AMP in P. monodon in response to the administration of various immunostimulants viz., two marine yeasts (Candida haemulonii S27 and Candida sake S165) and two bglucan isolates (extracted from C. haemulonii S27 and C. sake S165) were noted during the study. Responses to the application of two grampositive probiotic bacteria (Bacillus MCCB101 and Micrococcus MCCB104) were also observed. The immune profile was recorded preand postchallenge white spot syndrome virus (WSSV) by semiquantitative RTPCR. Expressions of seven WSSV genes were also observed for studying the intensity of viral infection in the experimental animals. The crustinlike AMP was found to be constitutively expressed in the animal and a significant downregulation could be noted postchallenge WSSV. Remarkable downregulation of the gene was observed in the immunostimulant fed animals prechallenge followed by a significant upregulation postchallenge WSSV. Tissuewise expression of crustinlike AMP on administration of C. haemulonii and Bacillus showed maximum transcripts in gill and intestine. The marine yeast, C. haemulonii and the probiotic bacteria, Bacillus were found to enhance the production of crustinlike AMP and confer significant protection to P. monodon against WSSV infection

"A Sequence in Subdomain 2 of DBL1a of Plasmodium falciparum Erythrocyte Membrane Protein 1 Induces Strain Transcending Antibodies"

Immunity to severe malaria is the first level of immunity acquired to Plasmodium falciparum. Antibodies to the variant antigen PfEMP1 (P. falciparum erythrocyte membrane protein 1) present at the surface of the parasitized red blood cell (pRBC) confer protection by blocking microvascular sequestration. Here we have generated antibodies to peptide sequences of subdomain 2 of PfEMP1-DBL1a previously identified to be associated with severe or mild malaria. A set of sera generated to the amino acid sequence KLQTLTLHQVREYWWALNRKEVWKA, containing the motif ALNRKE, stained the live pRBC. 50% of parasites tested (7/14) were positive both in flow cytometry and immunofluorescence assays with live pRBCs including both laboratory strains and in vitro adapted clinical isolates. Antibodies that reacted selectively with the sequence REYWWALNRKEVWKA in a 15-mer peptide array of DBL1a-domains were also found to react with the pRBC surface. By utilizing a peptide array to map the binding properties of the elicited anti-DBL1a antibodies, the amino acids WxxNRx were found essential for antibody binding. Complementary experiments using 135 degenerate RDSM peptide sequences obtained from 93 Ugandan patient-isolates showed that antibody binding occurred when the amino acids WxLNRKE/D were present in the peptide. The data suggests that the ALNRKE sequence motif, associated with severe malaria, induces strain-transcending antibodies that react with the pRBC surface

A sequence in subdomain 2 of DBL1 alpha of plasmodium falciparum erythrocyte membrane protein 1 induces strain transcending antibodies

Immunity to severe malaria is the first level of immunity acquired to Plasmodium falciparum. Antibodies to the variant antigen PfEMP1 (P. falciparum erythrocyte membrane protein 1) present at the surface of the parasitized red blood cell (pRBC) confer protection by blocking microvascular sequestration. Here we have generated antibodies to peptide sequences of subdomain 2 of PfEMP1-DBL1 alpha previously identified to be associated with severe or mild malaria. A set of sera generated to the amino acid sequence KLQTLTLHQVREYWWALNRKEVWKA, containing the motif ALNRKE, stained the live pRBC. 50% of parasites tested (7/14) were positive both in flow cytometry and immunofluorescence assays with live pRBCs including both laboratory strains and in vitro adapted clinical isolates. Antibodies that reacted selectively with the sequence REYWWALNRKEVWKA in a 15-mer peptide array of DBL1 alpha-domains were also found to react with the pRBC surface. By utilizing a peptide array to map the binding properties of the elicited anti-DBL1 alpha antibodies, the amino acids WxxNRx were found essential for antibody binding. Complementary experiments using 135 degenerate RDSM peptide sequences obtained from 93 Ugandan patient-isolates showed that antibody binding occurred when the amino acids WxLNRKE/D were present in the peptide. The data suggests that the ALNRKE sequence motif, associated with severe malaria, induces strain-transcending antibodies that react with the pRBC surface.

La desaparición forzada de personas: análisis crítico del derecho administrativo colombiano a través de los estándares del sistema interamericano de protección de derechos humanos. Estudio de caso de los mal llamados “falsos positivos”

La categorización jurídica de los mal llamados “falsos positivos” en Colombia es un tema controversial: en algunas ocasiones se habla de ejecuciones extrajudiciales, otras veces de homicidio en persona protegida y en otros casos de desapariciones forzadas, dependiendo de la institución que juzgue los casos. Ahora bien, al no existir unidad de criterios en la categorización jurídica de los “falsos positivos”, se hace imprescindible la integración de conceptos en torno a entender dichos hechos ilícitos inequívocamente como tales, toda vez que esto permitiría generar seguridad jurídica al interior del derecho administrativo colombiano. Precisamente, en lo a referente a la responsabilidad extracontractual del Estado. De igual manera, el concepto de reparación adoptado en los estamentos de Derecho Administrativo Colombiano, no siempre coincide con los estándares delineados por el Sistema interamericano de protección de Derechos Humanos, lo cual repercute de manera negativa en el tratamiento dado a las víctimas de los “falsos positivos” que encuadran en la categoría de desapariciones forzadas, lo que hace necesario el estudio detallado de las decisiones propias de cada sistema y compararlas, para buscar un mejoramiento de los estándares nacionales de reparación. Para dar un aporte teórico importante, esta investigación partirá del análisis sistemático de la Convención Interamericana sobre Desaparición Forzada de Personas, y la jurisprudencia de la Corte Interamericana de Derechos Humanos, y después inspeccionará los elementos fácticos y la reparación de los “falsos positivos”, determinando el grado de aplicación que se ha dado del sistema interamericano en los fallos proferidos por el Consejo de Estado.

The effect of temperature, photosynthetic photon flux density, and photoperiod on the vegetative growth and flowering of 'Autumn Bliss' raspberry

The effects of temperature, photosynthetic photon flux density (PPFD) and photoperiod on vegetative growth and flowering of the raspberry (Rubus idaeus L.) 'Autumn Bliss' were investigated. Increased temperature resulted in an increased rate of vegetative growth and a greater rate of progress to flowering. Optimum temperatures lay in the low to mid 20degreesC range. Above this the rate of plant development declined. Increased PPFD also advanced flowering. While photoperiod did not significantly affect the rate of vegetative growth, flowering occurred earliest at intermediate photoperiods and was delayed by extreme photoperiods. These responses suggest that there is potential for adjusting cropping times of raspberry grown under protection by manipulating the environment, especially temperature.

Evidence for the conspecificity of Palisada papillosa with P. perforata (Ceramiales, Rhodophyta) from the western and eastern Atlantic Ocean on the basis of morphological and molecular analyses

Morphological and molecular studies were carried out on Palisada papillosa and P. perforata from the Canary Islands (type locality of P. perforata), Mexico and Brazil. The two species have been distinguished by features of their external morphology such as size and degree of compactness of the thalli, presence or absence of arcuate branches, branching pattern and basal system. A detailed morphological comparison between these taxa showed that none of the vegetative anatomical or reproductive characters was sufficient to separate these species. The presence or absence of cortical cells in a palisade-like arrangement, also previously used to. distinguish these species, is not applicable. The species present all characters typical of the genus, and both share production of the first pericentral cell underneath the basal cell of the trichoblast, production of two fertile pericentral cells (the second and the third additional, the first remaining sterile), spermatangial branches produced from one of two laterals on the suprabasal cell of trichoblasts, and the procarpbearing segment with four pericentral cells. Details of the procarp are described for the species for the first time. The phylogenetic position of these species was inferred by analysis of the chloroplast-encoded rbcL gene sequences from 39 taxa, using one other Rhodomelacean taxon and two Ceramiaceae as outgroups. Relationships within the clade formed by P. papillosa and P. perforata have not been resolved due to the low level of genetic variation in their rbcL sequences (0-0.4%). Considering this and the morphological similarities, we conclude that P. papillosa is a taxonomic synonym of P. perforata. The phylogenetic analyses also supported the nomenclatural transfer of two species of Chondrophycus to Palisada, namely, P. patentiramea (Montagne) Cassano, Senties, Gil-Rodriguez & M.T. Fujii comb. nov. and P. thuyoides (Kutzing) Cassano, Senties, Gil-Rodriguez & M.T. Fujii comb. nov.

Efficiency of uniparental male and female care against egg predators in two closely related syntopic harvestmen

Although the benefits of maternal care have been investigated in many species, the caring role of males in species with exclusive paternal care has received less attention. We experimentally quantified the protective role of paternal care in the harvestman Iporangaia pustulosa. Additionally, we compared the effectiveness of paternal care against predation in this species with a syntopic harvestman with maternal care, Acutisoma proximum. We demonstrated that nearly one-third of the unprotected Iporangaia clutches disappeared entirely in 12 days, while the other two-thirds suffered a mean reduction of 55% in egg number. Conversely, 50% of the control clutches did not suffer any reduction, and only one was entirely consumed by predators. We also demonstrated that the mucus coat that covers Iporangaia clutches has an important deterrent role against predation by conspecifics: 58.3% of the clutches without mucus were attacked and three of them were entirely consumed, whereas only three clutches with mucus were attacked, suffering a reduction of up to three eggs. Iporangaia males were as efficient as Acutisoma females in protecting eggs. However, unattended Acutisoma eggs were attacked 20% more frequently than unattended Iporangaia eggs. Unattended Iporangaia eggs are protected by a mucus coat that prevents or decreases predation rate, whereas Acutisoma eggs are more susceptible to predation, probably because they lack this mucus coat. Thus, besides the fact that Iporangaia males efficiently protect the offspring against egg predators, females also contribute to egg protection by providing a mucus coat that deters egg predators. (C) 2009 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.

Occurrence and characterization of injuries caused by Mycotretus apicalis Lacordaire, 1842 (Coleoptera: Erotylidae) on cultivation of Pleurotus sajor-caju

When carrying out experiments on the production of the edible mushroom Pleurotus sajor-caju in the Laboratory of Edible Mushrooms, Universidade Federal de Lavras, Lavras, Brazil, in the second half of 2007, the presence of beetles later identified as belonging to the species Mycotretus apicalis was verified. This is the first recorded instance of this insect in cultures of P. sajor-caju in Brazil. The larvae and adults of this insect feed on the fruiting bodies of commercial harvests, resulting in reduction in mushroom quality. To provide evaluation of the injuries caused by these insects, substrates colonized by P. sajor-caju were infested with 4, 8, 16, 32 and 64 insects per block of substrate being the qualitative and quantitative losses then noted. Despite the lack of an observed decrease in biological efficiency, the injuries caused by these insects affected the commercial quality of the mushrooms, which may result in economic losses. The results showed that infestations of 32 insects per 0.8 kg of substrate led to a depreciation in the prices of mushrooms meant to be sold.

Gradual Decline in Malaria-Specific Memory T Cell Responses Leads to Failure to Maintain Long-Term Protective Immunity to Plasmodium chabaudi AS Despite Persistence of B Cell Memory and Circulating Antibody

The mechanisms responsible for the generation and maintenance of immunological memory to Plasmodium are poorly understood and the reasons why protective immunity in humans is so difficult to achieve and rapidly lost remain a matter for debate. A possible explanation for the difficulty in building up an efficient immune response against this parasite is the massive T cell apoptosis resulting from exposure to high-dose parasite Ag. To determine the immunological mechanisms required for long-term protection against P. chabaudi malaria and the consequences of high and low acute phase parasite loads for acquisition of protective immunity, we performed a detailed analysis of T and B cell compartments over a period of 200 days following untreated and drug-treated infections in female C57BL/6 mice. By comparing several immunological parameters with the capacity to control a secondary parasite challenge, we concluded that loss of full protective immunity is not determined by acute phase parasite load nor by serum levels of specific IgG2a and IgG1. Abs, but appears to be a consequence of the progressive decline in memory T cell response to parasites, which occurs similarly in untreated and drug-treated mice with time after infection. Furthermore, by analyzing adoptive transfer experiments, we confirmed the major role of CD4(+) T cells for guaranteeing long-term full protection against P. chabaudi malaria. The Journal of Immunology, 2008, 181: 8344-8355.

Attempts at a peptide vaccine against paracoccidioidomycosis, adjuvant to chemotherapy

Chemotherapy is the basis of treatment of paracoccidioidomycosis in its various forms. Depending on the Paracoccidioides brasiliensis virulence, the status of host immunity, the degree of tissue involvement and fungal dissemination, treatment can be extended for long periods with an alarming frequency of relapses. Association of chemotherapy with a vaccine to boost the cellular immune response seemed a relevant project not only to reduce the time of treatment but also to prevent relapses and improve the prognosis of anergic cases. The candidate immunogen is the gp43 major diagnostic antigen of P. brasiliensis and more specifically its derived peptide P10, carrying the CD4(+) T-cell epitope. Both gp43 and P10 protected Balb/c mice against intratracheal infections with virulent P. brasiliensis strain. P10 as single peptide or in a multiple-antigen-peptide (MAP) tetravalent construction was protective without adjuvant either by preimmunization and intratracheal challenge or as a therapeutic agent in mice with installed infection. P10 showed additive protective effects in drug-treated mice stimulating a Th-1 type immune response with high IFN-gamma and IL-12. P10 and few other peptides in the gp43 were selected by Tepitope algorithm and actually shown to promiscuously bind several prominent HLA-DR molecules suggesting that a peptide vaccine could be devised for a genetically heterogenous population. P10 was protective in animals turned anergic, was effective in a DNA minigene vaccine, and increased the protection by monoclonal antibodies in Balb/c mice. DNA vaccines and peptide vaccines are promising therapeutic tools to be explored in the control of systemic mycoses.

Balanoposthitis caused by Pseudomonas aeruginosa co-producing metallo-beta-lactamase and 16S rRNA methylase in children with hematological malignancies

Balanoposthitis is defined as the inflammation of the glans penis and its foreskin. In the presence of other underlying medical conditions, this localized infection may spread systemically, serving as a source of fever and bacteremia in neutropenic males. Two rare cases of balanoposthitis caused by a clonally related Pseudomonas aeruginosa isolate co-producing the SPM-1 metallo-beta-lactamase and the novel 16S rRNA methylase RmtD are described. Four multidrug-resistant (MDR) P. aeruginosa isolates were successively recovered from glans/foreskin swabs and urine cultures from two uncircumcised pediatric patients, one with Burkitt`s non-Hodgkin`s lymphoma and one with acute lymphoblastic leukemia. Clinically, preputial colonization by MDR P. aeruginosa evolved to severe balanoposthitis with glans/foreskin lesions as a source of fever. Combination therapy of ciprofloxacin and/or aztreonam (systemic) plus polymyxin B (topical) was effective once reversion of the neutropenic condition was achieved. Although P. aeruginosa remains an unusual cause of balanoposthitis, these cases should alert the physician to the potential pathogenicity of this bacterium. Furthermore, co-production of metallo-beta-lactamase and 16S rRNA methylase has a potential impact on the empirical management of complicated infections caused by P. aeruginosa. Crown Copyright (C) 2009 Published by Elsevier Ltd on behalf of International Society for Infectious Diseases. All rights reserved.

Survey of preys captured and used by Polybia platycephala (Richards) (Hymenoptera: Vespidae, Epiponini)

As vespas sociais são predadoras de várias espécies de insetos e, portanto, o estudo de suas presas pode revelar seu potencial para programas de controle biológico de pragas. Durante o período de setembro de 2000 a janeiro de 2002, foram realizadas 70h de coleta de presas capturadas em doze ninhos de Polybia platycephala Richards, localizados em áreas urbanas do município de Juiz de Fora, MG. As presas capturadas por P. platycephala compreenderam cinco ordens de insetos: Diptera (33,4%), Lepidoptera (28,6%), Hemiptera (12,0%), Hymenoptera (9,4%) e Coleoptera (7,2%). O peso médio da carga protéica transportada pelas vespas foi 1,9 ±1,6 mg (n = 34, 0,3 - 6,2 mg), e a taxa média de proteína transportada por dia foi 22,8 mg. de acordo com os resultados, pode-se estimar a captura de 4.380 presas por ano por uma única colônia de P. platycephala. Desta forma, a espécie pode ser utilizada em programas de manejo em ambientes urbanos contribuindo para o controle de insetos pragas como larvas de pernilongos, lagartas desfolhadoras de plantas de jardins, pulgões e formas aladas de formigas.

Concentração de nitrogênio uréico plasmático (NUP) e produção de leite de vacas mestiças mantidas em gramíneas tropicais sob pastejo rotacionado

O trabalho foi realizado com o objetivo de estudar os níveis de nitrogênio uréico plasmático (NUP) de vacas mestiças leiteiras mantidas em duas áreas de pastejo rotacionado, uma de capim-elefante cv. Guaçu (Pennisetum purpureum Schum. cv. Guaçu) e outra de capim-Tanzânia (Panicum maximum Jacq. cv. Tanzânia). Foram avaliadas a produção de forragem e as frações colmo, folha e material morto, com base na matéria seca (MS), bem como os teores de proteína bruta (PB) e fibra em detergente neutro (FDN) nas amostras de forragem. O sangue foi coletado em quatro épocas, utilizando-se 16 vacas por espécie de capim. A média da massa de forragem pré-pastejo foi de 5.321 e 5.384 kg de MS/ha por ciclo de pastejo para o capim-elefante e capim-Tanzânia, respectivamente. A forragem desaparecida pós-pastejo, referente à folha, ao colmo e ao material morto foi de 1.400, 620 e 443 kg de MS/ha para o capim-elefante e de 1.586, 736 e 132 kg de MS/ha para o capim-Tanzânia, por ciclo de pastejo. As médias de quatro ciclos de pastejo de PB e FDN nas amostras da planta inteira, colmo, folha, material morto e resíduo foram de 8,4; 78,5; 6,9; 77,1; 12,5; 76,3; 5,3; 82,6; 6,5; 80,2, respectivamente, para o capim-elefante, e de 9,3; 81,6; 7,4; 82,4; 14,3; 80,4; 6,3; 82,9; 7,6; 81,6, respectivamente, para o capim-Tanzânia. Foi encontrada diferença significativa nos níveis de NUP das vacas alimentadas nas diferentes espécies de capim e por época de amostragem do sangue. As médias de NUP foram 9,8 mg/dL para as vacas mantidas no capim-elefante e 10,6 mg/dL para as vacas mantidas no capim-Tanzânia. Os baixos níveis de NUP encontrados refletem a baixa ingestão de proteína bruta por vacas manejadas em pastagens tropicais.

Impactos do estresse salino e da cobertura morta nas características químicas do solo e no desenvolvimento do amaranto

Availability of good quality water has been reduced vertiginously, over the last decade, in the world. In some regions, the water resources have high concentration of the dissolved salts, these characteristics of the water make it s use impossible. Water quality can be a limitation for irrigated agriculture, principally in regions of arid or semiarid climate where the water resources are generally saline and are exposed at high evaporation ratio. For that reason, precipitation of the salts occurs near the soil surface and those salts themselves cumulate in the vegetal tissue, reducing the soil fertility and crop production. The adoption of tolerant crop to the water salinity and soil salinity, adaptable to the climatic conditions is other emergent necessity. This work had the goal of studying the effects of four salinity levels of the irrigation water salinity and use of mulch, dried leaves of Forest mangrove (Acacia mangiumWilld), in cultivated soil with amaranth (Amaranthus cruentus, BRS Alegria variety), in greenhouse. It was utilized the transplant of plants to PVC columns, containing 30 kg of silty loam soil, 10 days after emerging, with space of 50 x 50 cm between lines. Treatments were composed by combination of four levels of salinity (0.147; 1.500; 3.000 e 4.500 dS m-1), obtained by addition NaCl (commercial) to irrigation water and soil with and without protection, by mulch. A factorial system 4 x 2 was used with four repetitions, totalizing 32 parcels. The concentrations of nutrients in soil solution have been evaluated, in the dry matter of the vegetal tissue (roots, stem, leaves and raceme residue), at the end of the vegetative cycle. The use of soil protection reduced time for the beginning inflorescence of plants, at the same time, the increase of the salinity delayed this phase of amaranth development. The use of the mulch effectively increased the height, stem diameter, area of the larger leaf, humidity and dry matter content and amaranth grain production. The vegetal species showed salinity tolerance to experimented levels. The adopted treatments did not affect the pH values, exchangeable cation contents, electrical conductivity of soil solution (EC1:5) and saturated extract (ECSE), and Ca+2, Mg+, Fe+2 and Mn+2 contents, in the soil solution. The increase of the salinity concentration in the irrigation water inhibited the mineralization process of the organic matter (OM) and, consequently, the efficiency in the it´s utilization by plants, at the same time, produced increase in the values of the exchangeable sodium percentage (ESP), sodium adsorption ratio (SAR) and potassium adsorption ratio (PAR), in the soil solution. Therefore, the use of the mulch did not affect the first three parameters. The protein and nutrient contents: K+, Ca+2, P, Mg+2 e Cu+2, in amaranth grains, were improved by tillage condition. The raceme residues showed chemical/nutritional composition that makes advantageous its application in animal ration. In this context, it follows that amaranth tolerate the saline stress, of the irrigation water, until 4.500 dS m-1, temperature and relative humidity of the air predominant in the experimental environment

Induction of apoptosis in A549 pulmonary cells by two Paracoccidioides brasiliensis samples

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