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Citrus Huanglongbing (HLB), caused by the phloem-inhibiting bacterium Candidatus Liberibacter asiaticus (CLas), is the most devastating citrus disease, intimidating citrus production worldwide. Although commercially cultivated citrus cultivars are vulnerable to CLas infection, HLB-tolerant attributes have, however, been observed in certain citrus varieties, suggesting a possible pathway for identifying innate defense regulators that mitigate HLB. By adopting transcriptome and small RNAome analysis, the current study compares the responses of HLB-tolerant lemon (Citrus limon L.) with HLB-susceptible Shatangju mandarin (Citrus reticulata Blanco cv. Shatangju) against CLas infection. Transcriptome analysis revealed significant differences in gene expression between lemon and Shatangju. A total of 1751 and 3076 significantly differentially expressed genes were identified in Shatangju and lemon, respectively. Specifically, CLas infected lemon tissues demonstrated higher expressions of genes involved in antioxidant enzyme activity, protein phosphorylation, carbohydrate, cell wall, and lipid metabolism than Shatangju. Wet-lab experiments further validated these findings, demonstrating increased antioxidant enzyme activity in lemon: APX (35%), SOD (30%), and CAT (64%) than Shatangju. Conversely, Shatangju plants exhibited higher levels of oxidative stress markers like H2O2 (44.5%) and MDA content (65.2%), alongside pronounced ion leakage (11.85%), than lemon. Moreover, microscopic investigations revealed that CLas infected Shatangju phloem exhibits significantly more starch and callose accumulation than lemon. Furthermore, comparative sRNA profiles revealed the potential defensive regulators for HLB tolerance. In Shatangju, increased expression of csi-miR166 suppresses the expression of disease-resistant proteins, leading to inadequate defense against CLas. Conversely, reduced expression of csi-miR166 in lemon plants enables them to combat HLB by activating disease-resistance proteins. The above findings indicate that when infected with CLas, lemon exhibits stronger antioxidative activity and higher expression of disease-resistant genes, contributing to its enhanced tolerance to HLB. In contrast, Shatangju shows lower antioxidative activity, reduced expression of disease-resistant genes, significant ion leakage, and extensive callose deposition, possibly related to damage to plant cell structure and blockage of phloem sieve tubes, thereby promoting the development of HLB symptoms.

Contamination by acid drainage is an environmental problem in mineralized regions, especially in the surroundings of sulphide mines. The water rock interaction process involves the oxidative dissolution of sulphides, naturally or by mining activity, that generates acidity which, in turn, produces sulfate, and water pollution by sulphide-hosted metals. The particular geology of the Iberian Pyrite Belt (IPB) in the southwestern Iberian Peninsula creates the ideal conditions for such water contamination. Different water types were sampled at 28 locations across the entire IBP metallogenetic province. Anions, metals, and arsenic were analysed to assess the influence of acid drainage. The results demonstrate the sulfate nature of waters in the vicinity of mines. Other types of water in the IPB region are mainly mixed chloride and bicarbonate (river and groundwater) and mixed and sodium-bicarbonate (lakes) types. Water quality assessment indicated strong contamination of surface waters that are directly influenced by mine wastes. There is the additional concern that some lakes and groundwaters exhibit concentrations of potentially toxic elements (e.g. Al, As) that are above the regulatory limits established by the European Commission Water Framework Directive.

The Iberian Pyrite Belt (IPB) is a world-class metallogenic province with volcanogenic massive sulphide ore deposits. Most ore exploitation occurred since pre-Roman time, creating extensive galleries, wells, waste-dumps and pit lakes. These last structures are a concern for their potential environmental impact because they accumulate large volumes of mine water affected acid mine drainage. The present work classifies the pit lakes based on the surface water hydrochemistry. Using the Ficklin diagram for classification, pit lake waters vary from acid, high-metal to high-acid, extreme-metal and exhibit similarities with other pit lakes from the Spanish sector of the IPB.

Potentially toxic elements are mobilized in aquatic systems in solution or bounded to colloids of different sizes, which may include nanometer particles. The present work studies the distribution of elements (Al, Fe, Zn, Mn, Co, Cd, Ni, Cu, and As) between small (

No ano de 2000, realizou-se um levantamento epidemiológico em Paulínia, São Paulo, Brasil, com o objetivo de verificar a prevalência dos principais problemas de saúde bucal, utilizando o índice CPOD e os critérios de diagnóstico da Organização Mundial da Saúde. A amostra foi probabilística e totalizou 1.151 indivíduos nas diferentes faixas etárias. Em crianças de cinco anos de idade, o índice ceod foi de 1,90, com 54,2% de crianças sem experiência de cárie. Aos 12 anos o CPOD foi de 1,0, com 46,4% de crianças livres de cárie. A prevalência de fluorose nos escolares foi de 30,5%, e a maioria dessas crianças apresentou fluorose muito leve (22,9%). A prevalência de opacidades e hipoplasias foi de 9,1%. Entre os adultos examinados, o índice de cuidados foi de 55,4 e em média 21,3 dentes apresentaram experiência de cárie. Nos idosos o CPOD foi ainda mais elevado (29,50), sendo composto predominantemente por dentes extraídos (93,0 %).

OBJETIVO: Avaliar a resposta biológica que o oleato de etanolamina possa desencadear na parede de veias superficiais normais de cães. MÉTODOS: Utilizados 39 cães, sem raça definida, adultos, machos, com peso variando entre 10 a 18 kg, distribuídos de modo aleatório em três grupos: grupo 1, avaliados após 7 dias, grupo 2, 14 dias e grupo 3, 21 dias. O procedimento foi realizado em duas fases. A primeira constou da injeção de 2 ml do oleato de monoetanolamina a 5%, por punção única na veia cefálica do membro torácico do cão. A segunda, realizada 7, 14 e 21 dias após, constou da retirada da peça operatória, tendo sido executada em três tempos diferentes, conforme o grupo a que pertencia o animal. As veias contralaterais foram extraídas como controle. Para estudo histológico utilizaram-se os métodos de hematoxilina-eosina e tricrômio de Masson. RESULTADOS: A trombose venosa e a organização do trombo ocorreram em todos animais estudados. A recanalização do trombo não foi observada de modo estatisticamente significante, até 21 dias de exame. Encontrou-se lesão de túnica média, que não foi acompanhada de correspondente processo inflamatório. Na túnica adventícia este processo foi visto nos três períodos de tempo estudados. Depósitos de hemossiderina em fagócitos ocorreram aos 14 e 21 dias de experimento. Extravasamento de esclerosante foi observado somente na primeira semana de estudo. Material hialino fibrinóide foi encontrado aos 21 dias de experimento. CONCLUSÕES: O oleato de etanolamina em contato com a parede interna da veia superficial produziu trombose venosa, a qual se organizou em todos os casos, não se observando sua recanalização durante o tempo deste ensaio. Houve lesão da túnica média venosa em todos animais estudados, sem que houvesse processo inflamatório reativo nesse local. Na túnica adventícia venosa surgiu processo inflamatório, além de sinais de extravasamento do esclerosante, da presença de hemossiderina e de material hialino externos à veia.

Determinou-se a importância epidemiológica de dípteros Fanniidae na infestação de mosca-do-berne, por meio da identificação das espécies presentes, da determinação daquelas utilizadas por Dermatobia hominis na veiculação de seus ovos, bem como, pelo conhecimento da dinâmica populacional das espécies mais abundantes. Foram utilizadas cinco armadilhas iscadas com fígado bovino cru deteriorado e colocadas em uma mata ciliar margeada por uma área de pastagem com presença constante de bovinos. O estudo foi desenvolvido em uma área da Embrapa Gado de Corte, em Campo Grande, Estado de Mato Grosso do Sul, Brasil, localizada a 20º27'S e 54º37'W. A captura dos insetos foi realizada semanalmente durante o período de 09/08/1999 a 03/08/2000. Foi capturado um total de 40.629 moscas da família Fanniidae, pertencendo a cinco espécies do gênero Fannia: F. pusio, F. heydenii, F. bahiensis e F. longipila, e uma a ser identificada. A espécie mais freqüente foi F. pusio, com 63,20% do total capturado, seguida de F. heydenii, com 28,82%. Somente 0,44% do total de fêmeas de F. heydenii (45 exemplares) capturadas, principalmente nos meses de agosto e setembro, portavam ovos de D. hominis e o número médio, por indivíduo, foi de 15,98±7,13. Observaram-se ovos de D. hominis apenas na região abdominal dos vetores. F. heydenii predominou no período seco (maio a setembro) e início do período chuvoso do ano (outubro e novembro). O número de exemplares portando ovos de D. hominis foi maior no final do período seco do ano, o que explica a alta incidência deste parasito em bovinos nos meses de setembro e outubro.

Foram utilizados 960 pintos, metade machos e metade fêmeas, no período de 1 a 21 dias idade, com peso inicial médio de 42,5 e 42,1g, respectivamente. O delineamento experimental utilizado foi em blocos casualizados, com seis tratamentos, dois sexos, quatro repetições por sexo e vinte aves por unidade experimental. Os tratamentos consistiram de uma dieta basal deficiente em fósforo (0,150%), suplementada com fósforo disponível, provenientes do fosfato bicálcico (0,0; 0,10; 0,20; 0,30; 0,40; e 0,50%). As variáveis avaliadas foram ganho de peso (GP), consumo de ração (CR), conversão alimentar (CA), resistência dos ossos à quebra (ROQ), cinzas (CO) e fósforo no osso (PO). As exigências em fósforo disponível foram estimadas por intermédio dos modelos de regressão polinomial e LRP. As suplementações com fósforo disponível influenciaram o GP, CR, CA, ROQ, CO e PO, tanto para os machos como para as fêmeas. As estimativas de exigências de P-disp variaram de 0,259 (0,085%/Mcal de EM) a 0,559% (0,184%/Mcal de EM), para os machos, e de 0,307% (0,092%/Mcal de EM) a 0,575% (0,190%/Mcal de EM) para as fêmeas. Entretanto, considerando as respostas biológicas, para as variáveis de desempenho e características ósseas, pode-se sugerir que a exigência de fósforo disponível, para frangos de corte machos e fêmeas, de 1 a 21 dias de idade, é de 0,45 ou 0,148% de Pdisp/Mcal de EM.

Foram realizados dois experimentos para determinar o nível ideal de proteína bruta para frangos de corte Ross de ambos os sexos, nas fases inicial (1 a 21) e crescimento (22 a 42 dias de idade). Foram avaliados: o ganho de peso (GP), consumo de ração (CR), conversão alimentar (CA) e características de carcaça. Para cada fase, o delineamento experimental foi em blocos inteiramente casualisados. Na fase inicial, utilizou-se um fatorial 2x6, constituído de dois sexos e seis níveis de proteína bruta (20,00; 20,50; 21,00; 21,50; 22,00 e 22,50%), seis repetições e um total de 1440 aves. Na fase de crescimento, utilizou-se um fatorial 2x5, constituído de dois sexos e cinco níveis de proteína bruta (17,50; 18,00; 18,50; 19,00 e 19,50%), seis repetições e um total de 1200 aves. Os níveis de lisina total utilizados foram 1,27% e 1,16%, respectivamente para as fases inicial e de crescimento. Foram mantidas as relações lisina: metionina+cistina, treonina, triptofano e arginina, levando em consideração o perfil da proteína ideal. Com base nos resultados de desempenho, a exigência de proteína bruta estimada para machos, na fase de 1 a 21 dias foi de 22,42% PB, enquanto que para fêmeas, observou-se efeito linear para GP e CA. Na fase de crescimento, os machos responderam linearmente aos níveis de proteína para CR, CA, rendimento de peito e gordura abdominal, tendo estes resultados melhorados à medida que se aumentava o nível de proteína bruta da ração. Para as fêmeas, verificou-se efeito quadrático do nível dietético de proteína para GP e efeito linear para CA e gordura abdominal. Os níveis de proteína bruta estimados foram: na fase inicial 22,4 e 22,5% e na fase de crescimento 19,5 e 18,5% de PB, respectivamente, para machos e fêmeas.

Através de levantamentos empíricos, este artigo discute os estereótipos visuais propagados na cultura de consumo. Tratarei de alguns tópicos relativos às artes visuais, considerando que a visualidade contemporânea é intensamente marcada pelas imagens infantis veiculadas na mídia e por objetos de consumo. A influência do imaginário estereotipado constrói uma subjetividade homogeneizada, reforçando os discursos hegemônicos que estabelecem desigualdades. Pensando a arte como alternativa para novas formas de presentidade, defendo o desenvolvimento de um olhar crítico sobre estas imagens e aponto algumas estratégias para abordarmos esta questão dentro da sala de aula.

Duzentos leitões mestiços (Landrace × Large White) foram distribuídos em delineamento em blocos ao acaso, com quatro tratamentos e cinco repetições, com o objetivo de avaliar os efeitos de diferentes dietas sobre o desempenho e os pesos relativos e absolutos de fígado e pâncreas de leitões durante os períodos de aleitamento e pós-desmame, arraçoados dos 14 a 56 dias de idade. Os tratamentos consistiram de quatro dietas, as quais foram fornecidas aos animais dos 14 aos 42 dias de idade, quando então foram substituídas por uma dieta simples (ração de creche). Com base nos resultados, pode-se concluir que dietas contendo fontes de proteína de alta qualidade, como leite em pó integral e farinha de peixe, sem fonte suplementar de alimentos energéticos, como soro de leite, quando fornecidas a leitões dos 14 aos 42 dias de idade, proporcionaram os mesmos resultados de desempenho de leitões desmamados aos 28 dias de idade e recebendo rações simples à base de milho e farelo de soja.

Com o objetivo de determinar as exigências de metionina+cistina, foram utilizados 1440 e 1080 frangos de corte da marca comercial Hubbard, 50% de cada sexo, para as fases de crescimento e acabamento, respectivamente. Seis níveis de suplementação de DL-metionina (0,0; 0,06, 0,12; 0,18; 0,24; e 0,30%) foram adicionados às rações basais deficientes em metionina+cistina. Foram avaliados, aos 22-42 e 43-56 dias, ganho de peso, consumo de ração, conversão alimentar, rendimento e composição química da carcaça, gordura abdominal e concentração plasmática de ácido úrico. Com base nas análises estatística e nos parâmetros estudados durante a fase de crescimento (22 a 42 dias de idade), sugere-se utilizar 0,896 e 0,856% de metionina+cistina total para machos e fêmeas, respectivamente. Para a fase de acabamento (43 a 56 dias de idade), com base nas análises estatísticas e nos parâmetros estudados, sugere-se utilizar 0,764 e 0,740% de metionina+cistina total para machos e fêmeas, respectivamente.

Neste estudo observamos a reação tecidual após implantação de próteses de silicone associadas à poliglactina 910. Telas de borracha de silicone com 1,5 x 1,5cm de diâmetro e 0,7mm de espessura, polifenestradas, foram implantadas no subcutâneo do dorso de ratos.Dois grupos foram observados, no primeiro apenas telas fenestradas eram implantadas, no segundo as fenestrações das telas eram preenchidas com fios de poliglactina 910. Os animais foram observados por 30, 60 e 90 dias quando eram sacrificados e as telas mais o tecido adjacente ressecados. Não se observou diferenças na quantidade de fibras colágenas entre os subgrupos de um mesmo grupo e ou quando comparados com os subgrupos do grupo em estudo. No grupo controle, onde se usou apenas as telas fenestradas de silicone houve uma diferença significante entre as fibras reticulares, em maior número nos grupos de 30 e 60 dias de observação, quando comparadas ao grupo de 90 dias. A poliglactina 910 associada ao silicone não alterou o número de fibras do tecido conjuntivo, contadas nas fenestrações das telas.

OBJETIVO: Estudar a ultra-estrutura das vilosidades intestinais de ratos após diferentes tempos de clampeamento total do pedículo hepático. MÉTODOS: Foram utilizados 40 ratos, machos, divididos em quatro grupos de 10 animais cada um. O grupo Sham não foi submetido a isquemia. Os grupos E1, E2 e E3 sofreram isquemia de 10, 20 e 30 minutos, respectivamente. Ao final do experimento, fragmentos do intestino delgado (íleo) foram retirados e processados para microscopia eletrônica de transmissão. RESULTADOS: Nos grupos E1 e E2 observou-se infiltração de leucócitos, dilatação vascular e áreas eletrolúcidas na lâmina própria das vilosidades. Já no grupo E3 observou-se desprendimento do revestimento epitelial, desintegração das microvilosidades e núcleos em apoptose. Na lâmina própria notamos áreas hemorrágicas, vasos sangüíneos e linfáticos dilatados e inúmeros leucócitos. CONCLUSÕES: o clampeamento do pedículo hepático hepático provoca congestão esplâncnica, tempo dependente, sendo que aos 30 minutos se nota desprendimento das células epiteliais, apoptose e hemorragia.

O objetivo da pesquisa foi estudar a eficácia do enxerto autógeno de fáscia lata, para tratar ferimentos penetrantes provocados no esôfago cervical de cães. Foram estudadoscomparativamente dois procedimentos operatórios para se corrigir lesões orificiais provocadas na parede do esôfago cervical do cão com uma área padronizada de 3,12 cm2 Foram operados 40 cães, divididos em 2 grupos iguais denominados, grupo A (sutura ) e grupo B (enxerto) .Cada grupo foi subdividido em dois sub grupos denominados A1, A2 e B1, B2, com 10 animais cada para serem reoperados no 7° e no 30° P.O.respectivamente.No grupo controle, (A ) foi aplicada a sutura em plano único extramucosa com pontos simples de náilon 5-zeros .No grupo experimento (B) aplicou-se para ocluir o defeito provocado um enxerto de fáscia lata, suturado à borda da parede do esôfago com pontos simples extramucosa em plano único, com fio de náilon 5-zeros. Comparou-se os achados macroscópicos e microscópicos dos dois grupos não se observando diferença estatística significante, mas quanto ao aspecto clínico, os animais do grupo A apresentaram resultados melhores no que se refere a formação de fístulas apresentando um caso, enquanto o grupo B apresentou cinco casos Concluimos que a fáscia lata não é eficaz para tratar ferimentos penetrantes no esôfago cervical de cães.

Abstract Metabolic Syndrome (MetS) is associated with increased risk of morbi-mortality, thus the characterization of the population magnitude of this syndrome is critical for allocating health care. However, prevalence estimates of MetS in the same population could differ depending on the definition used. Therefore, we compared the prevalence of the MetS using definitions proposed by: National Cholesterol Education Panel Revised (NCEP) and International Diabetes Federation (IDF) 2009 in a Japanese-Brazilians community (131 individuals, age 57 ± 16 years, 1st and 2nd generation). All individuals went through a clinical and laboratorial evaluation for assessment of weigh, height, waist circumference, blood pressure, triglycerides, HDL-cholesterol and fasting plasma glucose. The prevalence of MetS was 26.7% (n = 35) and 37.4% (n = 49) under the NCEP and IDF definitions, respectively. Despite higher blood pressure measurements, waist circumference and serum triglyceride levels and lower HDL cholesterol levels (p

Abstract Background Bathymodiolus azoricus is a deep-sea hydrothermal vent mussel found in association with large faunal communities living in chemosynthetic environments at the bottom of the sea floor near the Azores Islands. Investigation of the exceptional physiological reactions that vent mussels have adopted in their habitat, including responses to environmental microbes, remains a difficult challenge for deep-sea biologists. In an attempt to reveal genes potentially involved in the deep-sea mussel innate immunity we carried out a high-throughput sequence analysis of freshly collected B. azoricus transcriptome using gills tissues as the primary source of immune transcripts given its strategic role in filtering the surrounding waterborne potentially infectious microorganisms. Additionally, a substantial EST data set was produced and from which a comprehensive collection of genes coding for putative proteins was organized in a dedicated database, "DeepSeaVent" the first deep-sea vent animal transcriptome database based on the 454 pyrosequencing technology. Results A normalized cDNA library from gills tissue was sequenced in a full 454 GS-FLX run, producing 778,996 sequencing reads. Assembly of the high quality reads resulted in 75,407 contigs of which 3,071 were singletons. A total of 39,425 transcripts were conceptually translated into amino-sequences of which 22,023 matched known proteins in the NCBI non-redundant protein database, 15,839 revealed conserved protein domains through InterPro functional classification and 9,584 were assigned with Gene Ontology terms. Queries conducted within the database enabled the identification of genes putatively involved in immune and inflammatory reactions which had not been previously evidenced in the vent mussel. Their physical counterpart was confirmed by semi-quantitative quantitative Reverse-Transcription-Polymerase Chain Reactions (RT-PCR) and their RNA transcription level by quantitative PCR (qPCR) experiments. Conclusions We have established the first tissue transcriptional analysis of a deep-sea hydrothermal vent animal and generated a searchable catalog of genes that provides a direct method of identifying and retrieving vast numbers of novel coding sequences which can be applied in gene expression profiling experiments from a non-conventional model organism. This provides the most comprehensive sequence resource for identifying novel genes currently available for a deep-sea vent organism, in particular, genes putatively involved in immune and inflammatory reactions in vent mussels. The characterization of the B. azoricus transcriptome will facilitate research into biological processes underlying physiological adaptations to hydrothermal vent environments and will provide a basis for expanding our understanding of genes putatively involved in adaptations processes during post-capture long term acclimatization experiments, at "sea-level" conditions, using B. azoricus as a model organism.

Realizou-se um estudo da ação dos fios de categute cromado 6-0 e de poliglecaprone 25, 5-0, para determinar o fio ideal na ileocistoplastia em ratas. O maior objetivo, foi a profilaxia litiásica. Utilizaram-se 51 ratos de Wistar, que participaram de dois grupos experimentais: grupo do plano piloto (27 ratos), que foi o grupo categute (GC) e grupo da tese do mestrado (24 ratos), que foi o grupo poliglecaprone (GP). Sob anestesia intraperitonial com pentobarbital sódico a 3%, todos os animais foram submetidos a ileocistoplastia, após laparotomia mediana longitudinal. Nos ratos do GC, a anastomose da bexiga urinária (aberta 0,5 cm sagitalmente no seu ápice), com a extremidade distal do segmento ileal ,era realizada por meio de pontos separados em plano único, com fio de categute cromado 6-0, e nas ratas do GP, com fio de poliglecaprone 25, 5-0. Seguiu-se a síntese da parede abdominal e recuperação anestésica. O estudo era realizado em 27, 42 e 57 dias nos animais do GC, que eram divididos em 3 subgrupos de 9 ratos. No GP, os animais eram analisados em 28 e 84 dias, por subgrupos de 12 animais. Transcorrido o tempo determinado para cada subgrupo, eram reoperados e observados os aspectos macroscópicos da cicatrização, aderências e formação de cálculos. A bexiga urinária ampliada pelo segmento ileal, era ressecada, aberta, lavada em solução salina isotônica, fixada no Líquido de Boüin, e processada , para histologia. A eutanásia era consumada com dose mínima letal anestésica. Os resultados apresentados, foram 29,6% de litíase, nas ratas do GC: 5,4% em 27dias; 10% em 42 dias e 14,25 em 57 dias do pós operatório (p.o.). Nas ratas do GP, não foram encontrados cálculos. Os estudos microscópicos, foram submetidos à análise estatística (a £ a 0,05), e utilizados para outro trabalho científico. Conclui-se: na ileocistoplastia com categute cromado 6-0, há 29,6% de cálculos, do 27o ao 57o dia p.o. e com o uso do fio de poliglecaprone 25, 5-0, até o 84o p.o.,não há presença de litíase.

Pollination is vital for ecosystem functioning, especially in biodiversity-rich regions like the Brazilian Cerrado. Our research establishes a comprehensive meta network of pollinator-plant interactions within this biome. We quantified the importance of different pollinator groups, identifying keystone species. We examined potential biases in sampling effort and the spatial behavior of interactions within the heterogeneous Cerrado plant physiognomies. Our investigation uncovered 1499 interactions among 293 plant species and 386 visitor species, with legitimate pollination accounting for 42.4% of the interactions. The network exhibited modularity, driven by bees and insects, with vertebrates bridging diurnal and nocturnal modules. While a generalized pattern emerged, high specialization existed within modules due to habitat diversity. Bees, particularly Apis mellifera (exotic) and Trigona spinipes (native), played central roles as network hubs. Hummingbirds and bats, engaged in specialized interactions showing strong connectivity within and between modules. Interestingly, invertebrate-vertebrate modules were more prevalent than expected in the meta network. However, a bias was evident, primarily within specific biogeographical districts with fragmented landscapes and intrusion from other biomes. Variations in plant species and endemism rates influenced pollinator occurrence and the Cerrado network topology. Our study offers valuable insights into pollinator-plant interactions within the Cerrado, encompassing both invertebrates and vertebrates. The modeled network represents a significant step in understanding the structural complexity of pollination networks, integrating partial networks from diverse pollination systems within heterogeneous habitats. Nevertheless, a biogeographical bias could limit a comprehensive understanding of network functionality across the Cerrado., Competing Interests: The authors have no conflict of interest., (© 2024 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Chronic stress and elevated levels of glucocorticoids (GCs), the main stress hormones, accelerate Alzheimer's disease (AD) onset and progression. A major driver of AD progression is the spreading of pathogenic Tau protein between brain regions, precipitated by neuronal Tau secretion. While stress and high GC levels are known to induce intraneuronal Tau pathology (i.e. hyperphosphorylation, oligomerization) in animal models, their role in trans-neuronal Tau spreading is unexplored. Here, we find that GCs promote secretion of full-length, primarily vesicle-free, phosphorylated Tau from murine hippocampal neurons and ex vivo brain slices. This process requires neuronal activity and the kinase GSK3β. GCs also dramatically enhance trans-neuronal Tau spreading in vivo, and this effect is blocked by an inhibitor of Tau oligomerization and type 1 unconventional protein secretion. These findings uncover a potential mechanism by which stress/GCs stimulate Tau propagation in AD., (© 2024. The Author(s).)

Background: Extracellular vesicles (EVs), including small EVs (sEVs) such as exosomes, exhibit great potential for the diagnosis and treatment of brain disorders, representing a valuable tool for precision medicine. The latter demands high-quality human biospecimens, especially in complex disorders in which pathological and specimen heterogeneity, as well as diverse individual clinical profile, often complicate the development of precision therapeutic schemes and patient-tailored treatments. Thus, the collection and characterization of physiologically relevant sEVs are of the utmost importance. However, standard brain EV isolation approaches rely on tissue dissociation, which can contaminate EV fractions with intracellular vesicles., Methods: Based on multiscale analytical platforms such as cryo-EM, label-free proteomics, advanced flow cytometry, and ExoView analyses, we compared and characterized the EV fraction isolated with this novel method with a classical digestion-based EV isolation procedure. Moreover, EV biogenesis was pharmacologically manipulated with either GW4869 or picrotoxin to assess the validity of the spontaneous-release method, while the injection of labelled-EVs into the mouse brain further supported the integrity of the isolated vesicles., Results: We hereby present an efficient purification method that captures a sEV-enriched population spontaneously released by mouse and human brain tissue. In addition, we tested the significance of the release method under conditions where biogenesis/secretion of sEVs was pharmacologically manipulated, as well as under animals' exposure to chronic stress, a clinically relevant precipitant of brain pathologies, such as depression and Alzheimer's disease. Our findings show that the released method monitors the drug-evoked inhibition or enhancement of sEVs secretion while chronic stress induces the secretion of brain exosomes accompanied by memory loss and mood deficits suggesting a potential role of sEVs in the brain response to stress and related stress-driven brain pathology., Conclusions: Overall, the spontaneous release method of sEV yield may contribute to the characterization and biomarker profile of physiologically relevant brain-derived sEVs in brain function and pathology. Video Abstract., (© 2023. The Author(s).)

The colloidal stability of non-motile algal cells in water drives their distribution in space. An accurate description of the interfacial properties of microalgae is therefore critical to understand how microalgae concentrations can change in their biotope or during harvesting processes. Here, we probe the surface charges of three unicellular algae─ Chlorella vulgaris , Nannochloropsis oculata , and Tetraselmis suecica ─through their electrophoretic mobility. Ohshima's soft particle theory describes the electrokinetic properties of particles covered by a permeable polyelectrolyte layer, a usual case for biological particles. The results appear to fit the predictions of Ohshima's theory, proving that all three microalgae behave electrokinetically as soft particles. This allowed us to estimate two characteristic parameters of the polyelectrolyte external layer of microalgae: the volume charge density and the hydrodynamic penetration length. Results were compared with transmission electron microscopy observations of the algal cells' surfaces, and in particular of their extracellular polymeric layer, which was identified with the permeable shell evidenced by electrophoretic measurements. Noticeably, the algal surface potentials estimated from electrophoretic mobility using the soft particle theory are less negative than the apparent zeta potentials. This finding indicates that electrostatics are expected to play a minor role in phenomena of environmental and industrial importance, such as microalgae aggregation or adhesion.

Insects are the most abundant and diverse organisms on Earth and provide essential ecosystem services. However, Brazilian society rarely consider the importance of insects in their diverse country. Therefore, in this review, we provide an overview of ecosystem services provided by insects in Brazil. A database search returned 136 articles, published in English or Portuguese, on ecosystem services provided by insects in Brazil. The first article was published in 1982, and majority of the studies were conducted in the Atlantic Forest or the Cerrado biomes. The most frequently studied insect-provided ecosystem services were pollination, decomposition, and biological control of pests. The studies focused primarily on natural and anthropic ecosystems, and most followed an experimental approach. We noted that the term "ecosystem services" was not used frequently in studies on insects in Brazil. The information available was mostly taxon-biased. We discuss the implications of these findings in relation to reconciling economic interests and the need for insect conservation for continued provision of ecosystem services in a broader perspective. In conclusion, we argue that the scientific community should focus on understanding the ecosystem services provided by insects other than those strictly related to economic activities, and on improving communication with policymakers and citizens. As a tropical and megadiverse country, Brazil has the potential to become a protagonist in conserving and using the ecosystem services provided by insects, both locally and internationally, by providing scientific information to policymakers and citizens.

Chagas disease and malaria are two neglected tropical diseases (NTDs) that prevail in tropical and subtropical regions in 149 countries. Chagas is also present in Europe, the US and Australia due to immigration of asymptomatic infected individuals. In the absence of an effective vaccine, the control of both diseases relies on chemotherapy. However, the emergence of parasite drug resistance is rendering currently available drugs obsolete. Hence, it is crucial to develop new molecules. Phthalimides, thiosemicarbazones, and 1,3-thiazoles have been used as scaffolds to obtain antiplasmodial and anti-Trypanosoma cruzi agents. Herein we present the synthesis of 24 phthalimido-thiosemicarbazones (3 a-x) and 14 phthalimido-thiazoles (4 a-n) and the corresponding biological activity against T. cruzi, Plasmodium falciparum, and cytotoxicity against mammalian cell lines. Some of these compounds showed potent inhibition of T. cruzi at low cytotoxic concentrations in RAW 264.7 cells. The most active compounds, 3 t (IC 50 =3.60 μM), 3 h (IC 50 =3.75 μM), and 4 j (IC 50 =4.48 μM), were more active than the control drug benznidazole (IC 50 =14.6 μM). Overall, the phthalimido-thiosemicarbazone derivatives were more potent than phthalimido-thiazole derivatives against T. cruzi. Flow cytometry assay data showed that compound 4 j was able to induce necrosis and apoptosis in trypomastigotes. Analysis by scanning electron microscopy showed that T. cruzi trypomastigote cells treated with compounds 3 h, 3 t, and 4 j at IC 50 concentrations promoted changes in the shape, flagella, and surface of the parasite body similar to those observed in benznidazole-treated cells. The compounds with the highest antimalarial activity were the phthalimido-thiazoles 4 l (IC 50 =1.2 μM), 4 m (IC 50 =1.7 μM), and 4 n (IC 50 =2.4 μM). Together, these data revealed that phthalimido derivatives possess a dual antiparasitic profile with potential effects against T. cruzi and lead-like characteristics., (© 2020 Wiley-VCH GmbH.)

A cost-effective optical instrument for continuous in-situ monitoring applications is presented. With a production cost in raw materials of 38 €, a power consumption of 300 A in sleep mode and 100 mA in active mode (5 ms reading), and a capacity to monitor turbidity and sedimentary displacement at eight different depths in the water column, the sensor was developed for sediment monitoring in coastal areas. Due to the extent and dynamics of the processes involved in these areas, observations require a wide spatial and temporal resolution. Each of the eight monitoring nodes uses one infrared backscatter channel, to estimate turbidity and sediment concentration, and one ultraviolet with one infrared transmitted light channels to distinguish organic/inorganic composition of the suspended material load. An in-lab calibration was conducted, using formazine to correlate turbidity with the electronic outputs of the instrument. An analysis of the influence of external light sources and correction techniques were performed. Moreover, an in-lab experiment was conducted to study the behaviour of the sensor-to-sediment transport, wash load and sediment accumulation. The device was deployed, with a water level sensor, in an estuarine area with high sediment dynamics. The monitoring data were analysed, showing the potential of the device to continuously monitor turbidity, sediment processes, and distinguish between organic and inorganic matter, at the different depths in the water column.

A cost-effective optical sensor for continuous in-situ monitoring of turbidity and suspended particulate matter concentration (SPM), with a production cost in raw materials less than 20 €, is presented for marine or fluvial applications. The sensor uses an infrared LED and three photodetectors with three different positions related to the light source-135º, 90º and 0º-resulting in three different types of light detection: backscattering, nephelometry and transmitted light, respectively. This design allows monitoring in any type of environment, offering a wide dynamic range and accuracy for low and high turbidity or SPM values. An ultraviolet emitter-receiver pair is also used to differentiate organic and inorganic matter through the differences in absorption at different wavelengths. The optical transducers are built in a watertight structure with a radial configuration where a printed circuit board with the electronic signal coupling is assembled. An in-lab calibration of the sensor was made to establish a relation between suspended particulate matter (SPM) or the turbidity (NTU) to the photodetectors' electrical output value in Volts. Two different sizes of seashore sand were used (180 µm and 350 µm) to evaluate the particle size susceptibility. The sensor was tested in a fluvial environment to evaluate SPM change during sediment transport caused by rain, and a real test of 22 days continuous in-situ monitoring was realized to evaluate its performance in a tidal area. The monitoring results were analysed, showing the SPM change during tidal cycles as well as the influence of the external light and biofouling problems.

Spinal cord haematoma, or haematomyelia, is a rare condition caused by several unusual disease processes. Traumatic events, such as spinal cord injury and surgery or procedures involving the spinal cord, are the most important causes of spinal cord haematoma. Rarely, it is associated with anticoagulation therapy. Irrespective of cause, spinal cord haematoma is considered a neurosurgical emergency and must be treated promptly in order to prevent neurological sequelae. The authors describe the case of a 69-year-old patient taking warfarin in the therapeutic range for a mechanic mitral valve, who developed chest pain with cervical and dorsal radiation, and experienced sudden paraparesis of the limbs. A CT of the spine confirmed haematomyelia. A high index of suspicion, prompt recognition and immediate intervention are essential to prevent major morbidity and mortality from intraspinal haemorrhage., Learning Points: This article reports an unusual presentation of spontaneous spinal haematoma, imposing the careful elaboration of differential diagnoses, which is very important in internal medicine.The description of this low-incidence case allows the scientific community to assist in approaching patients with similar symptoms.The lack of studies about the etiology and treatment of spontaneous spinal haematoma underlines the need for further studies and research in the area in order to increase the scientific evidence on the approach of these patients., Competing Interests: Conflicts of Interests: The Authors declare that there are no competing interests.

In this study we compared DNA barcode-suggested species boundaries with morphology-based species identifications in the amphipod fauna of the southern European Atlantic coast. DNA sequences of the cytochrome c oxidase subunit I barcode region (COI-5P) were generated for 43 morphospecies (178 specimens) collected along the Portuguese coast which, together with publicly available COI-5P sequences, produced a final dataset comprising 68 morphospecies and 295 sequences. Seventy-five BINs (Barcode Index Numbers) were assigned to these morphospecies, of which 48 were concordant (i.e., 1 BIN = 1 species), 8 were taxonomically discordant, and 19 were singletons. Twelve species had matching sequences (<2% distance) with conspecifics from distant locations (e.g., North Sea). Seven morphospecies were assigned to multiple, and highly divergent, BINs, including specimens of Corophium multisetosum (18% divergence) and Dexamine spiniventris (16% divergence), which originated from sampling locations on the west coast of Portugal (only about 36 and 250 km apart, respectively). We also found deep divergence (4%-22%) among specimens of seven species from Portugal compared to those from the North Sea and Italy. The detection of evolutionarily meaningful divergence among populations of several amphipod species from southern Europe reinforces the need for a comprehensive re-assessment of the diversity of this faunal group.

Snake venoms are natural sources of biologically active molecules that are able to act selectively and specifically on different cellular targets, modulating physiological functions. Thus, these mixtures, composed mainly of proteins and peptides, provide ample and challenging opportunities and a diversified molecular architecture to design and develop tools and agents of scientific and therapeutic interest. Among these components, peptides and small proteins play diverse roles in numerous physiological processes, exerting a wide range of pharmacological activities, such as antimicrobial, antihypertensive, analgesic, antitumor, analgesic, among others. The pharmaceutical and cosmetic industries have recognized the huge potential of these privileged frameworks and believe them to be a promising alternative to contemporary drugs. A number of natural or synthetic peptides from snake venoms have already found preclinical or clinical applications for the treatment of pain, hypertension, cardiovascular diseases and aging skin. A well-known example is captopril, whose natural peptide precursor was isolated from Bothrops jararaca snake venom, which is a peptide-based drug that inhibits the angiotensin-converting enzyme, producing an anti-hypertensive effect. The present review looks at the main peptides (natriuretic peptides, bradykinin-potentiating peptides and sarafotoxins) and low mass proteins (crotamine, disintegrins and three-Finger toxins) from snake venoms, as well as synthetic peptides inspired by them, describing their biochemical, structural and physiological features, as well as their applications as research tools and therapeutic agents., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.)

Cancer remains a high incidence and mortality disease, causing around 8.2 million of deaths in the last year. Current chemotherapy needs to be expanded, making research for new drugs a necessary task. Immune system modulation is an emerging concept in cancer cell proliferation control. In fact, there are a number of mechanisms underlying the role immune system plays in tumor cells. In this work, we describe the structural design, synthesis, antitumor and immunomodulatory potential of 31 new 1,3-thiazole and thiosemicarbazone compounds. Cisplatin was used as anticancer drug control. Cytotoxicity against J774A.1 macrophages and antitumor activity against HT-29 and Jurkat cells was determined. These 1,3-thiazole and thiosemicarbazone compounds not only exhibited cytotoxicity in cancer cells, but were able to cause irreversible cancer cell damage by inducing necrosis and apoptosis. In addition, these compounds, especially pyridyl-thiazoles compounds, regulated immune factors such as interleukin 10 and tumor necrosis factor, possible by directing immune system in favor of modulating cancer cell proliferation. By examining their pharmacological activity, we were able to identify new potent and selective anticancer compounds., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

Chagas disease is a tropical disease caused by the parasite Trypanosoma cruzi, which is endemic in Central and South America. Few treatments are available with effectiveness limited to the early (acute) stage of disease, significant toxicity and widespread drug resistance. In this work we report the outcome of a HTS-ready assay chemical library screen to identify novel, nontoxic, small-molecule inhibitors of T. cruzi. We have selected 50 compounds that possess hydrazone as a common group. The compounds were screened using recombinant T. cruzi (Tulahuen strain) expressing beta-galactosidase. A 3D quantitative structure-activity relationship (QSAR) analysis was performed using descriptors calculated from comparative molecular field analysis (CoMFA). Our findings show that of the fifty selected hydrazones, compounds LpQM-19, 28 and 31 displayed the highest activity against T. cruzi, leading to a selectivity index (SI) of 20-fold. The 3D-QSAR analysis indicates that a particular electrostatic arrangement, where electron-deficient atoms are aligned along the molecule main axis positively correlates with compound biological activity. These results provide new candidate molecules for the development of treatments against Chagas disease., (Copyright © 2016. Published by Elsevier Ltd.)

Chagas disease is a parasitic infection caused by protozoan Trypanosoma cruzi that affects approximately 6-7 million people worldwide. Benznidazole is the only drug approved for treatment during the acute and asymptomatic chronic phases; however, its efficacy during the symptomatic chronic phase is controversial. The present work reports the synthesis and anti-T. cruzi activities of a novel series of phthalimido-thiazoles. Some of these compounds showed potent inhibition of the trypomastigote form of the parasite at low cytotoxicity concentrations in spleen cells, and the resulting structure-activity relationships are discussed. We also showed that phthalimido-thiazoles induced ultrastructural alterations on morphology, flagellum shortening, chromatin condensation, mitochondria swelling, reservosomes alterations and endoplasmic reticulum dilation. Together, these data revealed, for the first time, a novel series of phthalimido-thiazoles-structure-based compounds with potential effects against T. cruzi and lead-like characteristics against Chagas disease., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

A main source of uncertainty currently associated with environmental risk assessment of chemicals is the poor understanding of the influence of environmental factors on the toxicity of xenobiotics. Aiming to reduce this uncertainty, here we evaluate the joint-effects of two pesticides (chlorpyrifos and mancozeb) on the terrestrial isopod Porcellionides pruinosus under different soil moisture regimes. A full factorial design, including three treatments of each pesticide and an untreated control, were performed under different soil moisture regimes: 25%, 50%, and 75% WHC. Our results showed that soil moisture had no effects on isopods survival, at the levels assessed in this experiment, neither regarding single pesticides nor mixture treatments. Additivity was always the most parsimonious result when both pesticides were present. Oppositely, both feeding activity and biomass change showed a higher sensitivity to soil moisture, with isopods generally showing worse performance when exposed to pesticides and dry or moist conditions. Most of the significant differences between soil moisture regimes were found in single pesticide treatments, yet different responses to mixtures could still be distinguished depending on the soil moisture assessed. This study shows that while soil moisture has the potential to influence the effects of the pesticide mixture itself, such effects might become less important in a context of complex combinations of stressors, as the major contribution comes from its individual interaction with each pesticide. Finally, the implications of our results are discussed in light of the current state of environmental risk assessment procedures and some future perspectives are advanced., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Bacterial colonization and subsequent biofilm formation is still one of the major problems associated with medical devices. Antimicrobial peptides (AMP) immobilization onto biomaterials surface is a promising strategy to avoid bacterial colonization. However, a correct peptide orientation and exposure from the surface is essential to maintain AMP antimicrobial activity. This work aims to evaluate the effect of the immobilization on antibacterial activity of Dhvar5 (LLLFLLKKRKKRKY), an AMP with a head-to-tail amphipathicity. Dhvar5 was linked to thin chitosan coatings in i) a controlled orientation and exposure, testing covalent immobilization of its N- or C-terminus and using spacers with different lengths and flexibilities or in ii) a random orientation by physical adsorption. Chitosan coating was chosen due to its antimicrobial properties and readiness to be functionalized. Surface characterization demonstrated the chemoselective immobilization of the peptide with different spacers in a similar concentration (∼2 ng/cm2). Efficacy assays demonstrated that covalent immobilization of Dhvar5 exposing its cationic end, improves the chitosan coating antimicrobial effect by decreasing Methicillin-resistant Staphylococcus aureus (MRSA) colonization. This effect was enhanced when longer spacers were used independently of their flexibility. In opposite, immobilized Dhvar5 exposing its hydrophobic end has no effect on bacterial adhesion to chitosan, and when adsorbed in a random orientation even induces bacterial adhesion to chitosan coating., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Chagas disease, caused by Trypanosoma cruzi, is a life-threatening infection leading to approximately 12,000 deaths per year. T. cruzi is susceptible to thiosemicarbazones, making this class of compounds appealing for drug development. Previously, the homologation of aryl thiosemicarbazones resulted in an increase in anti-T. cruzi activity in comparison to aryl thiosemicarbazones without a spacer group. Here, we report the structural planning, synthesis and anti-T. cruzi evaluation of new aryl thiosemicarbazones (9a-x), designed as more conformationally restricted compounds. By varying substituents attached to the phenyl ring, substituents were observed to retain, enhance or greatly increase the anti-T. cruzi activity, in comparison to the nonsubstituted derivative. In most cases, hydrophobic and bulky substituents, such as bromo, biphenyl and phenoxyl groups, greatly increased antiparasitic activity. Specifically, thiosemicarbazones were identified that inhibit the epimastigote proliferation and were toxic for trypomastigotes without affecting mouse splenocytes viability. The most potent anti-T. cruzi thiosemicarbazones were evaluated against cruzain. However, inhibition of this enzyme was not observed, suggesting that the compounds work through another mechanism. In addition, examination of T. cruzi cell death showed that these thiosemicarbazones induce apoptosis. In conclusion, the structural design executed within the series of aryl thiosemicarbazones (9a-x) led to the identification of new potent anti-T. cruzi agents, such as compounds (9h) and (9r), which greatly inhibited epimastigote proliferation, and demonstrated a toxicity for trypomastigotes, but not for splenocytes. Mechanistically, these compounds do not inhibit the cruzain, but induce T. cruzi cell death by an apoptotic process., (Copyright © 2014 Elsevier Masson SAS. All rights reserved.)

Schistosomiasis is a chronic and debilitating disease caused by a trematode of the genus Schistosoma and affects over 207 million people. Chemotherapy is the only immediate recourse for minimizing the prevalence of this disease and involves predominately the administration of a single drug, praziquantel (PZQ). Although PZQ has proven efficacy, there is a recognized need to develop new drugs as schistosomicides since studies have shown that repeated use of this drug in areas of endemicity may cause a temporary reduction in susceptibility in isolates of Schistosoma mansoni. Hydrazones, thiosemicarbazones, phthalimides, and thiazoles are thus regarded as privileged structures used for a broad spectrum of activities and are potential candidates for sources of new drug prototypes. The present study determined the in vitro schistosomicidal activity of 10 molecules containing these structures. During the assays, parameters such motility and mortality, oviposition, morphological changes in the tegument, cytotoxicity, and immunomodulatory activity caused by these compounds were evaluated. The results showed that compounds formed of thiazole and phthalimide led to higher mortality of worms, with a significant decline in motility, inhibition of pairing and oviposition, and a mortality rate of 100% starting from 144 h of exposure. These compounds also stimulated the production of nitric oxide and tumor necrosis factor alpha (TNF-α), thereby demonstrating the presence of immunomodulatory activity. The phthalyl thiazole LpQM-45 caused significant ultrastructural alterations, with destruction of the tegument in both male and female worms. According to the present study, phthalyl thiazole compounds possess antischistosomal activities and should form the basis for future experimental and clinical trials.

Objectives: To assess the prevalence of anemia among children attending public preschools in Ilhabela, state of São Paulo, Brazil, and their response to a treatment regimen., Methods: Between 2007 and 2008, the weight, height, and hemoglobin levels of 667 children were measured in the school setting. Anemic children were prescribed a 12-week course of ferrous sulfate, and follow-up assessment visits were scheduled for halfway through this course and the end of treatment., Results: The prevalence of anemia was 25.6% (N = 170). The lowest hemoglobin level measured was 9.5 g/dL. Anemia was most prevalent (36%) in children between the ages of 60 and 65 months. Median hemoglobin levels rose to 11.8 g/dL from 10.5 g/dL after treatment; 76% of children recovered from anemia., Conclusion: Screening for anemia in the school setting and prompt therapy, including mid-treatment follow-up, proved to be an effective strategy for facing this major public health issue.

Shiga-like toxin 2 (Stx2)-producing enterohemorrhagic Escherichia coli (referred to as EHEC or STEC) strains are the primary etiologic agents of hemolytic-uremic syndrome (HUS), which leads to renal failure and high mortality rates. Expression of Stx2 is the most relevant virulence-associated factor of EHEC strains, and toxin neutralization by antigen-specific serum antibodies represents the main target for both preventive and therapeutic anti-HUS approaches. In the present report, we describe two Salmonella enterica serovar Typhimurium aroA vaccine strains expressing a nontoxic plasmid-encoded derivative of Stx2 (Stx2DeltaAB) containing the complete nontoxic A2 subunit and the receptor binding B subunit. The two S. Typhimurium strains differ in the expression of flagellin, the structural subunit of the flagellar shaft, which exerts strong adjuvant effects. The vaccine strains expressed Stx2DeltaAB, either cell bound or secreted into the extracellular environment, and showed enhanced mouse gut colonization and high plasmid stability under both in vitro and in vivo conditions. Oral immunization of mice with three doses of the S. Typhimurium vaccine strains elicited serum anti-Stx2B (IgG) antibodies that neutralized the toxic effects of the native toxin under in vitro conditions (Vero cells) and conferred partial protection under in vivo conditions. No significant differences with respect to gut colonization or the induction of antigen-specific antibody responses were detected in mice vaccinated with flagellated versus nonflagellated bacterial strains. The present results indicate that expression of Stx2DeltaAB by attenuated S. Typhimurium strains is an alternative vaccine approach for HUS control, but additional improvements in the immunogenicity of Stx2 toxoids are still required.

To develop long-lasting, topical pour-on insecticides for dogs to control zoonotic visceral leishmaniasis, two deltamethrin-based formulations (emulsifiable concentrate [EC] and suspension concentrate [SC]) were tested for their efficacy against the phlebotomine sandfly Lutzomyia longipalpis Lutz & Neiva (Diptera: Psychodidae), vector of Leishmania infantum Nicolle (Kinetoplastida: Trypanosomatidae). The entomological outcomes tested were anti-feeding effect (proportion of female sandflies unfed), lethal effect (24-h female sandfly mortality) and these two effects combined, and the insecticide persistence time at 50% (residual activity, RA50) and 80% (RA80) efficacy. On initial application, the proportions of female flies that demonstrated anti-feeding activity or were killed were similar for both formulations, at 0.93 (95% confidence interval [CI] 0.856-0.977) vs. 0.81 (95% CI 0.763-0.858) (anti-feeding) and 0.86 (95% CI 0.787-0.920) vs. 0.76 (95% CI 0.698-0.817) (24-h mortality) for EC and SC, respectively. The RA(50) rates for anti-feeding and mortality caused by the EC formulation were 4.7 months (95% CI 4.18-5.84) and 2.5 months (95% CI 2.25-2.90), respectively, compared with 1.1 months (95% CI 0.96-1.15) and 0.6 months (95% CI 0.50-0.61), respectively, for the SC formulation. The RA(50) for the combined anti-feeding and mortality effects of EC was 5.2 months (95% CI 4.73-5.96), compared with only 0.9 months (95% CI 0.85-1.00) for the SC formulation. The four- to six-fold superior residual activity of the EC formulation was attributed to the addition of a solvent-soluble resin in the formulation which improved fur adhesion and acted as a reservoir for the slow release of the active ingredient. These results identify the potential of such a low-cost formulation to reduce the inter-intervention interval to 5-6 months, similar to that recommended for deltamethrin-impregnated dog collars or for re-impregnation of conventional bednets, both of which are currently used to combat Leishmania transmission. Finally, a novel bioassay was developed in which sandflies were exposed to fur from treated dogs, revealing no detectable tolerance (24-h mortality) in wild-caught sandflies to the insecticide formulations up to 8 months after the initiation of communitywide application of the insecticides to dogs.

No effective vaccine or immunotherapy is presently available for patients with the hemolytic uremic syndrome (HUS) induced by Shiga-like toxin (Stx) produced by enterohaemorragic Escherichia coli (EHEC) strains, such as those belonging to the O157:H7 serotype. In this work we evaluated the performance of Bacillus subtilis strains, a harmless spore former gram-positive bacterium species, as a vaccine vehicle for the expression of Stx2B subunit (Stx2B). A recombinant B. subtilis vaccine strain expressing Stx2B under the control of a stress inducible promoter was delivered to BALB/c mice via oral, nasal or subcutaneous routes using both vegetative cells and spores. Mice immunized with vegetative cells by the oral route developed low but specific anti-Stx2B serum IgG and fecal IgA responses while mice immunized with recombinant spores developed anti-Stx2B responses only after administration via the parenteral route. Nonetheless, serum anti-Stx2B antibodies raised in mice immunized with the recombinant B. subtilis strain did not inhibit the toxic effects of the native toxin, both under in vitro and in vivo conditions, suggesting that either the quantity or the quality of the induced immune response did not support an effective neutralization of Stx2 produced by EHEC strains.

The entomological efficacy of using 25% deltamethrin EC insecticide-treated bednets (ITNs) was evaluated against the sandfly Lutzomyia longipalpis Lutz and Neiva (Diptera: Psychodidae), the principal vector of zoonotic visceral leishmaniasis (ZVL) in Latin America. A crossover field study in Amazon Brazil (Marajó Island, Pará State) demonstrated that, compared with untreated nets, the insecticide increased the barrier effect of nets by 39% (95% confidence interval [CI] 34-44%), reduced human landing rates by 80% (95% CI 62-90%) and increased the 24-h mortality rate from 0% to 98% (95% CI 93-99%) inside ITNs. The presence of an ITN also reduced the human landing rate on unprotected persons outside the net in the same room by 56% (95% CI 52-59%), and increased 24-h mortality to 68% (95% CI 62-73%) compared to 0.4% (0.1-2.0%) in untreated houses. The reduction in human landing rates in ITN rooms was associated with a doubling in the proportion of sandflies alighting on walls compared with that in untreated rooms, which was attributed to insecticide-induced excito-repellency. There was no evidence that sandflies were diverted onto unprotected hosts. Human landing catches inside houses peaked between 19.00 hours and 23.00 hours and declined steadily to zero at 02.00 hours and thereafter. House-to-house questionnaires established that only 34% of households owned at least one net (median two, range 1-8), only 20% of the population slept under a net (33% of 0-5-year-old children), and the majority (73%) of the population slept in hammocks. Combined data pertaining to sleeping times for children and sandfly activity period indicate that > 50% of sandfly bites inside houses, and substantially more outside houses, were taken before a third of children were potentially protected by a net. This study demonstrates the clear entomological efficacy of ITNs against Lu. longipalpis in this endemic region. The effectiveness of ITNs at preventing ZVL infection and disease has still to be evaluated.

Electric field stimulation is widely used for heart pacing and arrhythmia reversion. In this study, we analysed the influence of waveform and direction of external stimulating electric field on the excitation threshold of isolated ventricular myocytes. The threshold field (E(T)) was lower when the field was applied longitudinally (E(T,L)) rather than transversally (E(T,T)) to the cell major axis. Rheobase was greater for transversal stimulation, but chronaxie and estimated membrane polarization were similar for both directions. The calculated maximal variation in membrane potential at the threshold (DeltaV(T) approximately 15 mV) was insensitive to field direction. As DeltaV(T) values were similar, we assumed that the E(T,T)/E(T,L) ratio might be described solely as the ratio of the major and minor cell semi-axes. Accordingly, the ratio thus estimated was comparable to that determined experimentally. Stimulus waveform significantly affected both E(T) and DeltaV(T), which were greater for monophasic versus biphasic stimuli. Direction and waveform effects were independent. We conclude that (a) direction affects E(T) by its influence on the ability of a given field intensity to cause threshold membrane polarization and (b) threshold-lowering effects of longitudinal stimulation and biphasic waveforms apparently depend on different mechanisms, are additive and thus may be combined to decrease the energy requirement for myocardial stimulation.

The occurrence of human pulmonary dirofilariasis maintains a relation with the prevalence of Dirofilaria immitis infection in the canine population. Several mosquito species are vectors of this nematode. Canine blood samples collected in Pingo d'Agua and União villages, Salvaterra municipality (Marajó Island, Pará), in June, 2004 (n = 34) and April, 2005 (N = 90) were analyzed. Parasitological and immunological (ELISA--kit SNAP(R) 3DX Biobrasil) diagnoses were compared following the examination of 34 samples. The prevalence in the population (N = 90) was evaluated by means of ELISA. ELISA revealed more positive samples (25/34; 73.5%) than thick smears (23/34, 67.6%) or Knott (21/34, 61.8%), but the differences were not significant (p > 0.05). The frequency of D. immitis infection was 58% in dogs ranging from 0-2 years old, whereas in older dogs it was 100%. The prevalence of canine dirofilariasis was high in Pingo d'Agua and Vila União (53.5%), indicating the risk of parasite transmission to the people in this area.

Among the causes for sudden death in epilepsy, cardiac dysfunction has been an area of interest. Based on this, the aim of our study was to evaluate the heart rate (in vivo and in vitro) and ventricular pressure in vitro of rats with epilepsy induced by pilocarpine. Adult male Wistar rats (n=6) were given pilocarpine hydrochloride to induce status epilepticus. Control rats (n=6) received saline solution instead pilocarpine. Our results showed significant differences in the mean of heart rate in vivo between the groups. In contrast, we did not find differences during in vitro experiments. Our results suggest a central nervous system modulation on the heart, which could explain the sudden unexpected death in epilepsy.

Atmospheric concentrations of volatile organic compounds were measured at two rural sites in central Portugal. The sites were chosen to be in line with the summer northwesterly sea breezes in order to study the evolution of the chemical composition of air masses during transport to inland areas. The most abundant non-oxygenated hydrocarbon in the ambient air was isoprene and the monoterpenes alpha-pinene, beta-pinene and 1,8 cineol. The maximum isoprene levels (6-7 ppb) were recorded at the most inland site, suggesting an enrichment of coastal air masses with biogenic emissions during transport over eucalyptus forests. Formaldehyde was the most prominent carbonyl compound in the atmosphere but acetaldehyde and acrolein were also abundant. Concentrations of carbonyl compounds had a tendency to be higher inland, particularly for glyoxal, methyl glyoxal, methyl vinyl ketone, metacrolein and pentanal. The observed increases indicate that carbonyls were produced by photochemical oxidation of biogenic hydrocarbons in aged air masses with coastal origin. Isoprene, monoterpenes and various carbonyls exhibited pronounced diurnal variations, which are explained on the basis of emissions from vegetation, oxidation pathways of biogenic hydrocarbons and meteorological conditions.

A primate model of leishmaniasis was developed with the objective of future vaccine testing. Lesion development and immunological parameters were studied upon primary and secondary infections. Seven Cebus apella were injected subcutaneously with 2 x 10(6) Leishmania (Leishmania) amazonensis promastigotes. Erythematous nodules appeared 19-29 days p.i., which disappeared 100 days p.i. Four months later, six of the monkeys were challenged with the same inoculum; three of them developed erythematous nodules after 7 days p.i., with ulcer formation in two of these subjects. The lesions were short-lived and all were cured 40 days post challenge. Anti-Leishmania IgG antibodies were detected and they increased after the challenge infection. Leishmania antigen-induced lymphoproliferation was found 1 month post-primary infection, which coincided with IFN-gamma production and lesion development. It decreased to control levels afterwards, but at the time of the challenge dose, it was significantly above the initial level. After the challenge infection, it first increased then decreased sharply at 40 days post-challenge, coinciding with the healing of the lesion. It increased again to a higher level at 60 days post-challenge. Leishmania (Leishmania) amazonensis-infection in C. apella did not induce complete protection against a secondary infection with a homologous parasite although specific antibody production and lymphoproliferation with IFN-gamma production were observed. This fact indicates that vaccine has to be better than infection in the induction of protective immunity, and raises a question on in vitro parameters that should be considered as a counterpart of expected protection induced by vaccine candidate. In addition, we conclude that this is a useful primate model for the evaluation of candidate vaccines.

Introduction: After birth, cardiac myocytes undergo substantial growth and differentiation that affect their stimulation threshold. Cells from younger animals have a higher stimulation threshold than cells from adults. The aim of this work was to compare the excitability of isolated hearts of rats during postnatal development by measuring the stimulation threshold at several pulse durations., Methods and Results: Stimulation threshold of isolated hearts were measured at eight different pulse durations between 0.1 to 20 msec. For each heart, a strength-duration curve was constructed and data were fitted using both Weiss-Lapicque and Blair models. Analysis of variance showed significant age-dependent differences in both rheobase field (E(reob)) and chronaxie (c). E(reob) decreased while c increased during development (E(reob) was 0.21, 0.16, 0.13, 0.10, and 0.09 V/cm and c was 2.0, 2.2, 2.3, 2.7, and 3.2 msec for rats aged 1, 2, 4, 8, and 20 weeks, respectively). There was a decrease in the threshold field with heart weight between 0.1 and 0.7 g, whereas the threshold was almost constant in the range from 0.7 to 2.0 g. Estimation of the energy density needed to defibrillate the heart was performed and appeared to be higher for younger than for adult animals., Conclusion: Hearts from younger animals have higher stimulation threshold than those from adults. This probably is due to changes in the cellular threshold as a result of maturation. The smaller excitability of younger hearts can have consequences with regard to the energy levels required for younger patients.

Studies on cardiac cell response to electric field stimulation are important for understanding basic phenomena underlying cardiac defibrillation. In this work, we used a model of a prolate spheroidal cell in a uniform external field (Klee and Plonsey, 1976) to predict the threshold electric field (ET) for stimulation of isolated ventricular myocytes of rats at different ages. The model assumes that ET is primarily determined by cell shape and dimensions, which markedly change during postnatal development. Neonatal cells showed very high ET, which progressively decreased with maturation (experimental mean values were 29, 21, 13, and 5.9 and 6.3 V/cm for 3-6, 13-16, 20-21, 28-35, and 120-180 day-old rats, respectively, P < 0.001; theoretical values were 24, 18, 11, 9, and 6 V/cm, respectively). Estimated maximum membrane depolarization at threshold (deltaVT approximately equals 35 mV, under our experimental conditions) was reasonably constant during development, except for cells from 1-mo-old animals, in which deltaVT was lower than at other ages. We conclude that the model reasonably correlates ET with cell geometry and size in most cases. Our results might be relevant for the development of efficient procedures for defibrillation of pediatric patients.