Abstract Background Insects can be found in numerous diverse environments, being exposed to pathogenic organisms like fungi and bacteria. Once these pathogens cross insect physical barriers, the innate immune system operates through cellular and humoral responses. Antimicrobial peptides are small molecules produced by immune signaling cascades that develop an important and generalist role in insect defenses against a variety of microorganisms. In the present work, a cecropin B-like peptide (AgCecropB) sequence was identified in the velvetbean caterpillar Anticarsia gemmatalis and cloned in a bacterial plasmid vector for further heterologous expression and antimicrobial tests. Methods AgCecropB sequence (without the signal peptide) was cloned in the plasmid vector pET-M30-MBP and expressed in the Escherichia coli BL21(DE3) expression host. Expression was induced with IPTG and a recombinant peptide was purified using two affinity chromatography steps with Histrap column. The purified peptide was submitted to high-resolution mass spectrometry (HRMS) and structural analyses. Antimicrobial tests were performed using gram-positive (Bacillus thuringiensis) and gram-negative (Burkholderia kururiensis and E. coli) bacteria. Results AgCecropB was expressed in E. coli BL21 (DE3) at 28°C with IPTG 0.5 mM. The recombinant peptide was purified and enriched after purification steps. HRMS confirmed AgCrecropB molecular mass (4.6 kDa) and circular dichroism assay showed α-helix structure in the presence of SDS. AgCrecropB inhibited almost 50% of gram-positive B. thuringiensis bacteria growth. Conclusions The first cecropin B-like peptide was described in A. gemmatalis and a recombinant peptide was expressed using a bacterial platform. Data confirmed tertiary structure as predicted for the cecropin peptide family. AgCecropB was capable to inhibit B. thuringiensis growth in vitro.

O objetivo do estudo foi avaliar o crescimento corporal e idade de início da puberdade em cordeiras sob duas dietas específicas para taxas de crescimento baixas e altas. Um rebanho de 20 cordeiras Bergamácia Brasileira foi dividido em dois grupos (n= 10) e mantido confinado em todo período experimental, sendo dispostos dois animais do mesmo tratamento/baia. Duas fases foram estabelecidas: Fase 1, a partir de 90 dias de idade até o início da puberdade, e Fase 2, a partir do início da puberdade até 1 ano de idade. Para a fase 1, duas dietas distintas foram formuladas, sendo: Tratamento A, formulada para obter um ganho de peso diário de aproximadamente 150g; e Tratamento B, para um ganho de peso médio diário de cerca de 250g. Na Fase 2, uma dieta equilibrada idêntica foi fornecida para ambos os grupos. A cada 14 dias, os animais foram pesados e calcularam-se o ganho médio diário, o consumo médio diário de matéria seca e escore de condição corporal. A partir do quinto mês de idade, em cada grupo, um macho vasectomizado foi usado para detectar estro, estabelecendo a idade de início da puberdade e estro para cada cordeira. As amostras de sangue foram coletadas a cada 28 dias para determinar a concentração plasmática de hormônio do crescimento. Cordeiras do tratamento B ganharam mais peso e tiveram maior escore de condição corporal (P<0,05), mas não houve diferença de idade para o início da puberdade e para os níveis plasmáticos de hormônio do crescimento (P>0,05). Verificou-se que os tratamentos A e B apresentaram desempenhos satisfatórios. Assim, o tratamento A pode ser indicado como um sistema de alimentação para alcançar respostas positivas em cordeiras submetidas ao confinamento durante a fase de crescimento.

The objective of the study was to evaluate body growth and age at onset of puberty on lambs fed two specific diets for low and high growth rates. A herd of 20 Brazilian Bergamasca lambs was divided in two groups (n= 10) and kept confined throughout the experimental period, two animals of the same treatment/pen. Two phases were established: Phase 1, from 90 days of age until the onset of puberty; and Phase 2, from puberty onset up to 1 year old. For Phase 1, two distinct diets were formulated, being: Treatment A, which was formulated to obtain an average daily gain of approximately 150g; and Treatment B, for an average daily gain of about 250g. In Phase 2, a balanced, equal diet was provided to both groups. Every 14 days, the animals were weighed and given average daily gain, average daily dry matter intake and body condition score. From the 5th month of age on, in each group, a vasectomized male was used to detect estrus, establishing age at puberty onset and estrus interval for each lamb. Blood samples were collected every 28 days to determine plasma growth hormone concentration. Treatment B lambs gained more weight and had higher body condition score (P<0.05) and there was no difference for age at puberty onset and plasma growth hormone levels (P>0.05) between treatments. It was found that both treatments showed satisfactory performances. Thus, treatment A may be indicated as a reasonable feeding system to achieve positive responses on confined ewe lambs during growth phase.

Rabies is a zoonotic disease that affects all mammals and leads to more than 55,000 human deaths every year, caused by rabies virus (RABV) (Mononegavirales: Rhabdoviridae: Lyssavirus). Currently, human rabies treatment is based on the Milwaukee Protocol which consists on the induction of coma and massive antiviral therapy. The aim of this study was to assess the decrease in the titer of rabies virus both in vitro and in vivo using short-interfering RNAs. To this end, three siRNAs were used with antisense strands complementary to rabies virus nucleoprotein (N) mRNA. BHK-21 cells monolayers were infected with 1000 to 0.1 TCID50 of PV and after 2 hours the cells were transfected with each of tree RNAs in separate using Lipofectamine-2000. All three siRNAs reduced the titer of PV strain in a least 0.72 logTCID50/mL and no cytotoxic effect was observed in the monolayers treated with Lipofectamine-2000. Swiss albino mice infected with 10.000 to 1 LD of PV strain by the intracerebral route were also transfected after two hours of infection with a pool 3 siRNAs with Lipofectamine-2000 by the intracerebral route, resulting in a survival rate of 30% in mice inoculated with 100 LD50, while the same dose led to 100% mortality in untreated animals. Lipofectamine-2000 showed no toxic effect in control mice. These results suggest that intracerebral administration of siRNAs might be an effective antiviral strategy for rabies.

The puma is an iconic predator that ranges throughout the Americas, occupying diverse habitats. Previous phylogeographic analyses have revealed that it exhibits moderate levels of genetic structure across its range, with few of the classically recognized subspecies being supported as distinct demographic units. Moreover, most of the species' molecular diversity was found to be in South America. To further investigate the phylogeographic structure and demographic history of pumas we analyzed mtDNA sequences from 186 individuals sampled throughout their range, with emphasis on South America. Our objectives were to refine the phylogeographic assessment within South America and to investigate the demographic history of pumas using a coalescent approach. Our results extend previous phylogeographic findings, reassessing the delimitation of historical population units in South America and demonstrating that this species experienced a considerable demographic expansion in the Holocene, ca. 8,000 years ago. Our analyses indicate that this expansion occurred in South America, prior to the hypothesized re-colonization of North America, which was therefore inferred to be even more recent. The estimated demographic history supports the interpretation that pumas suffered a severe demographic decline in the Late Pleistocene throughout their distribution, followed by population expansion and re-colonization of the range, initiating from South America.

INTRODUÇÃO: Complicações tromboembólicas são importantes fatores de risco para perda do enxerto e pior evolução após o transplante renal. Pacientes com defeito trombofílico apresentam maior risco de complicações tromboembólicas. Foram analisados, entre receptores de transplante renal, a prevalência de defeito trombofílico e o risco atribuído a esta condição para a perda do enxerto e para o desenvolvimento de tromboses intravasculares. MÉTODOS: Estudo do tipo coorte incluindo 388 receptores adultos analisados quanto à presença de trombofilia de acordo com a pesquisa de anticorpos anticardiolipina (aCL) por ELISA e das mutações G1691A no gene do fator V (FV) e G20210A no gene da protrombina (PT) por PCR multiplex. RESULTADOS: Defeito trombofílico foi identificado em 25,8% dos pacientes. As taxas de sobrevida de 2 anos do enxerto foram semelhantes entre os pacientes com e sem defeito trombofílico (94% versus 94%, p = 0,53), bem como a sobrevida dos enxertos livres de tromboses intravasculares (97% versus 97%, p = 0,83). Pacientes com defeito trombofílico apresentaram prevalência de tromboses intravasculares semelhante à do grupo-controle (3% versus 3,5%, p = 0,82). O transplante renal anterior foi associado a maior risco de perda de enxerto (OR 20,8, p < 0,001) e de ocorrência de tromboses intravasculares (OR 6,8, p = 0,008). CONCLUSÕES: As prevalências das mutações FV G1691A e PT G20210A na população estudada foram semelhantes às da população geral não transplantada, e a prevalência de anticorpos aCL superou a observada entre os indivíduos sadios. Não houve associação entre os marcadores de trombofilia estudados e a sobrevida em médio prazo do transplante renal.

INTRODUCTION: Thromboembolic complications are important risk factors for graft failure and worse renal transplantation outcome. Patients with thrombophilic disorders have a higher risk of thromboembolic complications. The prevalence of thrombophilic disorders and the associated risk for graft failure and for intravascular thrombosis were analyzed in renal transplant recipients. METHODS: This is a cohort study of 388 adult recipients investigated regarding the presence of thrombophilia, through the search for anticardiolipin antibodies (aCL) via ELISA and FV G1691A and PT G20210A gene mutations by multiplex PCR. RESULTS: Thrombophilic disorders were identified in 25.8% of the patients. The 2-year graft survival was similar among patients with and without thrombophilic disorder (94% versus 94%, p = 0.53), and so was the survival free of intravascular thrombosis (97% versus 97%, p = 0.83). The prevalence of intravascular thrombosis was similar in both groups (3% versus 3.5%, p = 0.82). Patients with previous kidney transplantation had a higher risk of graft failure (OR 20.8, p < 0.001) and of intravascular thrombosis (OR 6.8, p = 0.008). CONCLUSIONS: The prevalences of FV G1691A and PT G20210A gene mutations in this cohort of patients were similar to those of the general non-transplanted population. The prevalence of aCL antibodies was higher in this cohort than that observed in healthy individuals. The thrombophilic markers studied did not predict the medium-term survival of renal transplant.

We describe the experimental apparatus and the methods to achieve Bose-Einstein condensation in 87Rb atoms. Atoms are first laser cooled in a standard double magneto-optical trap setup and then transferred into a QUIC trap. The system is brought to quantum degeneracy selectively removing the hottest atoms from the trap by radio-frequency radiation. We also present the main theoretical aspects of the Bose-Einstein condensation phenomena in atomic gases.

OBJETIVO: Analisar a aplicabilidade do retalho fáscio-cutâneo em V-Y da região posterior da coxa na reparação de lesões isoladas ou múltiplas nas regiões glútea e perineal. MÉTODO: Foram submetidos a tratamento cirúrgico 20 pacientes, portadores de 25 úlceras nas regiões glútea e perineal, sendo 23 delas úlceras por pressão, um pós-abscesso perianal e um pós-trauma perineal. Em todos os pacientes a reparação foi realizada com o retalho fáscio-cutâneo da região posterior da coxa. O retalho foi confeccionado com base súpero-lateral, preservando-se os ramos fáscio-cutâneos das artérias glútea inferior, primeira e segunda perfurantes no tratamento de úlceras isquiáticas. Nas associações de úlcera isquiática com úlcera sacral, adicionou-se ao retalho uma extensão fáscio-cutânea da região glútea para tratamento cirúrgico em tempo único. Foi realizado retalho com base súpero-medial, preservando-se o ramo fáscio-cutâneo da artéria glútea inferior no tratamento das úlceras trocantéricas. Nas associações com úlcera sacral, acrescentou-se ao retalho posterior da coxa uma extensão fáscio-cutânea da região glútea, que permitiu o fechamento de todas as úlceras em um só tempo cirúrgico. Nas associações de úlceras trocantéricas e isquiáticas, realizou-se o retalho com base superior, preservando-se o ramo fáscio-cutâneo da artéria glútea inferior. RESULTADOS: Não houve necrose do retalho. As complicações imediatas foram três infecções, uma deiscência e um hematoma. Num período de seguimento de 6 meses a 29 meses, houve recidiva de seis úlceras (6/24 = 25,0%) em cinco pacientes (5/19 = 26,32%). Um paciente foi perdido neste seguimento. CONCLUSÃO: Conclui-se que o retalho fáscio-cutâneo posterior da coxa, em V-Y, pode ser utilizado com segurança no tratamento de lesões isoladas ou múltiplas das regiões glútea e perineal.

PURPOSE: To analyze the clinical applicability of one-stage treatment of single or multiple sores using the V-Y posterior thigh fasciocutaneous flap. METHODS: Twenty patients were treated, having 25 ulcers in the gluteal and perineal regions, being 23 pressure ulcers, 1 perianal abscess and 1 perineal trauma. In all of the patients the repair was carried out with a fasciocutaneous flap from the posterior region of the thigh. The flap was made with a superior and lateral base, preserving the fasciocutaneous branches of the inferior gluteal arteries, first and second perforating, in the treatment of ischial ulcers. In the associations of the same with sacral ulcer, a fasciocutaneous extension from the gluteal region was added to the flap for a one and only surgical procedure. A superior and medial based flap was made, preserving the fasciocutaneous branch of the inferior gluteal artery in the treatment of the trochanteric ulcers. In the associations with sacral ulcer a fasciocutaneous extension from the gluteal region was added to the posterior thigh flap which permitted the closure of all ulcers in only one surgical procedure. In the associations of trochanteric and ischial ulcers a flap with a superior base was made, preserving the fasciocutaneous branch of the inferior gluteal artery. RESULTS: There was no necrosis of the flap. The immediate complications were 3 infections, 1 dehiscences and 1 hematoma. In a follow-up period of 6 months to 29 months, with one patient being lost to follow up, there were 6 recurrences 6/24 (25.0%) in 5 patients 5/19 (26.32%). CONCLUSION: We conclude that the posterior fasciocutaneous thigh flap, in V-Y, can be used with safety in the treatment of isolated or multiple ulcers in the gluteal and perineal regions.

Realizou-se inquérito sorológico para malária em escolares de Marabá — Pará, por meio de testes de imunofluorescência (IP) para anticorpos IgG e IgM, tendo como antígenos P. falciparum e P. gallinaceum, e teste de hemaglutinação (HAg) com P. gallinaceum. O teste IF-IgG com P. falciparum foi positivo em 6,94% dos 389 indivíduos estudados e o de P. gallinaceum em 11,56%, havendo concordância entre ambos os testes em 88,68% das amostras. No total, observou-se 14,91% de casos reagentes em qualquer dos testes. O teste com P. gallinaceum se mostrou mais abrangente provavelmente devido a maior prevalência na região de infecções por P. vivax. Ao se dividir a população estudada em faixas etárias de 6 a 10 anos (grupo A) e de 11 a 16 anos (grupo B), observou-se diferença significativa de reatividade ao teste IF-IgG com P. falciparum (2,68% para A e 10,94% para B) mas não com P. gallinaceum (10,10% para A e 12,97% para B). Para os testes IF-IgM houve positividade de 2,83% na população, e para o teste de HAg de 1,80%, sem diferença significativa entre os grupos etários A e B.

A malaria serologic survey was carried out in first grade school children from Marabá, PA. Immunofluorescence (IF) test was used for the detection of IgG and IgM antibodies to P. falciparum and to P. gallinaceum. Also, hemagglutination (HAg) test with P. gallinaceum infected erythrocytes was employed. The IF-IgG test with P. falciparum was positive in 6.94% of the 389 individuals studied, and the IF-IgG test with P. gallinaceum in 11.56%. Agreement between both tests 'was observed in 88.68% of sera. A total 14.91% reactive cases were seen by any of the two tests. The IF test using P. gallinaceum as antigen showed higher positivity than P. falciparum probably due to the predominance of T. vivax infection in that area. A statistically significant difference (P < 0.05) was observed for IF-IgG test with P. falciparum antigen between the age group A (6 to 10 years) and group B (11 to 16 years). However, this was not seen when P. gallinaceum antigen was applied. The positivity obtained in IF-IgM and HAg tests were respectively 2.83% and 1.80%, with no significant differences between the referred age groups (P > 0.05).

Os valores de complemento hemolítico total, C3 total (nativo + produtos de degradação) e o grau de conversão de C3 nativo foram estudados em dois subgrupos de pacientes chagásicos, nas formas cardíaca e indeterminada, e em um subgrupo de indivíduos não chagásicos, clinicamente sadios. Os níveis de C3 total e as taxas de conversão de C3 em seus produtos de degradação foram semelhantes nos três subgrupos. Os valores de complemento hemolítico total foram estatisticamente diferentes nos três subgrupos (nível de significância descritivo p = 0,0757), tendo sido observada média aritmética mais baixa no subgrupo de cardíacos e mais elevada no subgrupo de controles. Maior amplitude de variação dos níveis de complemento hemolítico total foi notada no subgrupo de cardíacos, no qual se encontraram os valores extremos (máximo e mínimo), considerando-se todos os subgrupos.

The levels of hemolytic complement and total C3 (native C3 plus its degration products) and the degree of conversion of native C3 into its breakdown products were studied in sera from two subgroups of chagasic patients (indeterminate and cardiac forms) and from non chagasic individuals (control subgroup). The levels of total C3 and the degree of conversion of native C3 into its breakdown products were similar in the three subgroups. The levels of hemolytic complement were statistically different among the three subgroups. The lowest average was observed in the subgroup of cardiac patients and the highest average in the control subgroup of non chagasic individuals. The widest variation on levels of hemolytic complement was observed in the subgroup of cardiac patients in which we found the maximum and minimum values among all the subgroups.

O efeito da adubaçao NPK e da matéria orgânica na composição mineral do grão e na qualidade da bebida (prova de xícara) foi estudado. Verificou-se que a adubação fosfatada e o uso da matéria orgânica não influiram nas características mencionadas. A adubação nitrogenada e potássica aumentou o teor dos elementos correspondentes no grão e prejudicou pouco mas, significativamente a qualidade da bebida. Esses dados devem ser encarados com cautela e tidos apenas como preliminares.

In this second year of work, research was carried out concerning, the effect of N, P, K and organic fertilization on the mineral composition of the coffee bean and in the quality of the beverage. The material employed was obtained from a NxPxK, 2³ , factorial experiment with split plot (with and without organic matter). The test area is located in poor, deep and, sandy soil; Coffea Arabica L., Var. "Bourbon Vermelho" is the plant source. The main findings are as follows: a) N content in the bean was raised by the application of N in the soil. The use of K increases the level of K in the bean. The level of total P and soluble P was not affected by P fertilization. No treatment affected the S level in the bean. The level of the elements analysed was not affected significantly by the organic matter applied to the soil. b) Fruits collected in treatments where N and K were used gave significantly lower quality beverage. The quality of beverage wasn't affected by the organic matter. c) It was found a negative and significant (1%) correlation between N level in the bean and quality of beverage (r = -0,642). For K the correlation was found to be r - -0,449, significant at 10% of probability. No similar correlation was found with respect to P.

Foram feitas determinações dos teores de N, P e K no solo e nos frutos de cafeeiros submetidos a um ensaio fatotorial 2x2x2. Verificou-se que apenas as aplicações de P e K elevaram os teoresdos elementos no solo. O uso de N e K aumentou o teor dêsses nutrientes no fruto. A qualidade da bebida foi prejudicada pela falta de P na adubação.

This paper gives some preliminary results from determinations of the effect of N, P and K fertilization on the chemical composition of soil and coffee berries, as well in the quality of the beverage there of. The main findings are as follows: a) The use of P arid K caused an increase in the level of the respective element in the soil; no similar effect of N fertilisation was ascertained. b) N content in the bean tuas raised by that of K; P increased N content in the pulp. The application of P failed to raise its level in the fruit; the use of N, however decreased P % in the pulp. The use of KCl increased the level of K in the fruit. c) Fruits collected in treatments where P was omitted gave significantly lower quality beverage. A positive correlation was found between P content in the soil and quality of the beverage; the meaning - if any - of such correlation cannot be evaluated at present.