Transcription Factors Tec1 and Tec2 Play Key Roles in the Hyphal Growth and Virulence of Mucor lusitanicus Through Increased Mitochondrial Oxidative Metabolism.

Mucormycosis is a lethal and difficult-to-treat fungal infection caused by fungi of the order Mucorales. Mucor lusitanicus, a member of Mucorales, is commonly used as a model to understand disease pathogenesis. However, transcriptional control of hyphal growth and virulence in Mucorales is poorly understood. This study aimed to investigate the role of Tec proteins, which belong to the TEA/ATTS transcription factor family, in the hyphal development and virulence of M. lusitanicus. Unlike in the genome of Ascomycetes and Basidiomycetes, which have a single Tec homologue, in the genome of Mucorales, two Tec homologues, Tec1 and Tec2, were found, except in that of Phycomyces blakesleeanus, with only one Tec homologue. tec1 and tec2 overexpression in M. lusitanicus increased mycelial growth, mitochondrial content and activity, expression of the rhizoferrin synthetase-encoding gene rfs, and virulence in nematodes and wax moth larvae but decreased cAMP levels and protein kinase A (PKA) activity. Furthermore, tec1- and tec2-overexpressing strains required adequate mitochondrial metabolism to promote the virulent phenotype. The heterotrimeric G beta subunit 1-encoding gene deletant strain (Δgpb1) increased cAMP-PKA activity, downregulation of both tec genes, decreased both virulence and hyphal development, but tec1 and tec2 overexpression restored these defects. Overexpression of allele-mutated variants of Tec1(S332A) and Tec2(S168A) in the putative phosphorylation sites for PKA increased both virulence and hyphal growth of Δgpb1. These findings suggest that Tec homologues promote mycelial development and virulence by enhancing mitochondrial metabolism and rhizoferrin accumulation, providing new information for the rational control of the virulent phenotype of M. lusitanicus.

The heterotrimeric G-protein beta subunit Gpb1 controls hyphal growth under low oxygen conditions through the protein kinase A pathway and is essential for virulence in the fungus Mucor circinelloides.

Mucor circinelloides, a dimorphic opportunistic pathogen, expresses three heterotrimeric G-protein beta subunits (Gpb1, Gpb2 and Gpb3). The Gpb1-encoding gene is up-regulated during mycelial growth compared with that in the spore or yeast stage. gpb1 deletion mutation analysis revealed its relevance for an adequate development during the dimorphic transition and for hyphal growth under low oxygen concentrations. Infection assays in mice indicated a phenotype with considerably reduced virulence and tissue invasiveness in the deletion mutants (Δgpb1) and decreased host inflammatory response. This finding could be attributed to the reduced filamentous growth in animal tissues compared with that of the wild-type strain. Mutation in a regulatory subunit of cAMP-dependent protein kinase A (PKA) subunit (PkaR1) resulted in similar phenotypes to Δgpb1. The defects exhibited by the Δgpb1 strain were genetically suppressed by pkaR1 overexpression, indicating that the PKA pathway is controlled by Gpb1 in M. circinelloides. Moreover, during growth under low oxygen levels, cAMP levels were much higher in the Δgpb1 than in the wild-type strain, but similar to those in the ΔpkaR1 strain. These findings reveal that M. circinelloides possesses a signal transduction pathway through which the Gpb1 heterotrimeric G subunit and PkaR1 control mycelial growth in response to low oxygen levels.

Development and characterization of novel chimeric monoclonal antibodies for broad spectrum neutralization of rabies virus.

Current post-exposure prophylaxis for rabies virus infection has several limitations in terms of supply, cost, safety, and efficacy. Attempts to replace human or equine rabies immune globulins (HRIG or ERIG) have been made by several companies and institutes. We developed potent monoclonal antibodies to neutralize a broad spectrum of rabies viruses by screening hybridomas received from the U.S. Centers for Disease Control and Prevention (CDC). Two kinds of chimeric human antibodies (chimeric #7 and #17) were constructed by cloning the variable regions from selected hybridomas and the constant region of a human antibody. Two antibodies were bound to antigenic site III and I/IV, respectively, and were able to neutralize 51 field isolates of rabies virus that were isolated at different times and places such as Asia, Africa, North America, South America, and Australia. These two antibodies neutralize rabies viruses with high efficacy in an in vivo test using Syrian hamster and mouse models and show low risk for adverse immunogenicity.

Effects of dietary Forsythia suspensa on feed utilization, rumen fermentation, and immune response of Korean native goats (Capra hircus) / Efectos de la adición dietaria de Forsythia suspensa sobre utilización del alimento, fermentación ruminal y respuesta inmune en cabras nativas coreanas (Capra hircus) / Efeitos na dieta pela utilização de Forsythia suspensa nos alimentos, fermentação ruminal e resposta imune em caprinos nativos coreanos (Capra hircus)

Background: plant extracts of Forsythia suspensa, which contain pinoresinol, have been proposed as an alternative to antibiotics due to their antioxidant, anti-inflammatory, and microflora modulating effects. Objective: to evaluate the effects of dietary F. suspensa on feed utilization, rumen fermentation, blood parameters and immune response of Korean native goats (Capra hircus). Methods: a total of nine Korean native goats were assigned to one of three dietary treatments: 1) a basal diet without F. suspensa, 2) a basal diet added with 0.25% F. suspensa, or 3) a basal diet added with 0.50% F. suspensa. A change-over design was used in three consecutive periods to give 9 replicates per treatment. Each period lasted 18 days, including 15 days of adaptation to feed and environmental conditions and three days of total collection of feces and urine. Rumen fluid and blood samples were also collected. Nutrient digestibility, nitrogen retention, ruminal content, and blood metabolites—including immunoglobulins—were measured. Results: F. suspensa supplementation had no effect on nutrient digestibility, whole body nitrogen retention rate, ruminal pH, acetate, propionate, isobutyrate, butyrate, isovalerate, or valerate content (p>0.05). However, F. suspensa supplementation decreased total volatile fatty acids (VFA) concentration compared with the control diet, regardless of F. suspensa concentrations (p<0.05). Goats fed a diet containing 0.25% F. suspensa had greater alkaline phosphatase (ALP) concentration than those fed a control or a diet with 0.50% F. suspensa (p<0.05). Feeding different concentrations of dietary F. suspensa did not influence plasma IgA and IgM levels (p>0.05), although goats receiving both 0.25% and 0.50% F. suspensa diets had greater plasma IgG than those fed the control diet (p<0.05). Conclusion: supplementation of 0.25% F. suspensa significantly decreased total VFA concentrations and increased plasma IgG in Korean native goats, compared with those fed the control diet.

Antecedentes: los extractos de la planta Forsythia suspensa contienen pinoresinol. Dicha planta se ha propuesto como una alternativa a los antibióticos debido a su contenido de antioxidantes, antiinflamatorios y sus efectos moduladores de la microflora. Objetivo: evaluar los efectos dietarios de F. suspensa sobre la utilización del alimento, fermentación ruminal, parámetros sanguíneos y respuesta inmune en cabras nativas coreanas (Capra hircus). Métodos: un total de nueve cabras nativas coreanas fueron asignadas a 1 de 3 tratamientos: (1) una dieta basal sin F. suspensa, (2) una dieta basal con 0,25%, o (3) 0,50% de F. suspensa dietaría en un diseño change-over por tres periodos consecutivos con nueve réplicas por tratamiento. Cada periodo se prolongó por 18 días, incluyendo 15 días de adaptación al alimento y a las condiciones ambientales, y tres días de colección total de heces y orina. Tambien se colectó fluido ruminal y muestras sanguineas. Se midió la digestibilidad de nutrientes, nitrógeno retenido, contenido ruminal y metabolitos sanguíneos —incluyendo inmunoglobulinas—. Resultados: la suplementación con F. suspensa no tuvo efecto sobre la digestibilidad de los nutrientes, la tasa de retención de N, pH ruminal, ni sobre los contenidos de acetato, propionato, isobutirato, butirato, isovalerato o valerato (p>0,05). Sin embargo, la suplementación con F. suspensa disminuyo la concentración total de ácidos grasos volátiles (VFA) en comparacion con la dieta control, sin tener en cuenta las concentraciones de F. suspensa (p<0,05). Las cabras alimentadas con la dieta de 0,25% de F. suspensa tuvieron mayor concentración de fosfatasa alcalina (ALP) que cuando fueron alimentadas con la dieta control o con la de 0,50% de F. suspensa (p<0,05). Aunque la alimentación con diferentes concentraciones de F. suspensa no influyo en los niveles de IgA y IgM en plasma (p>0,05), las cabras que recibieron 0,25 y 0,50% de F. suspensa tuvieron mayor concentración de IgG en plasma que aquellas alimentadas con la dieta control (p<0,05). Conclusiones: la suplementación de 0,25% de F. suspensa disminuyo significativamente la concentración total de VFA e incrementó la IgG en el plasma de las cabras nativas coreanas, en comparación con aquellas que consumieron la dieta control.

Antecedentes: a Forsythiae suspensa é uma planta e seu extrato contém pinoresinol. Tem sido proposta como uma alternativa aos antibióticos existentes, devido ao seu teor de antioxidantes, anti-inflamatórias e efeitos moduladores da microflora, com base em estudos feitos in vivo e in vitro. Objetivo: avaliar os efeitos na dieta da inclusão de F. suspensa no aproveitamento alimentar, fermentação ruminal, parâmetros sanguíneos e resposta imune em caprinos coreanos nativos (Capra hircus). Métodos: um total de nove cabras coreanas nativas foram assignadas a um de três tratamentos na dieta: (1) uma dieta basal sem F. suspensa, (2) uma dieta basal com 0,25% de F. suspensa, ou (3) com 0,50% de F. suspensa na dieta. Se fez um desenho change-over por três períodos consecutivos com nove repetições por tratamento. Cada período de pesquisa consistiu de 18 dias, incluindo 15 dias de adaptação às condições alimentares e ambientais, e três dias de coleta total de fezes e urina. Também foram coletadas amostras de líquido ruminal e sangue. Mediou-se a digestibilidade dos nutrientes, o nitrogênio (N) retido, o conteúdo ruminal, e os parâmetros sanguíneos —incluindo imunoglobulinas—. Resultados: a suplementação com F. suspensa não teve nenhum efeito sobre a digestibilidade dos nutrientes, a taxa de retenção de N, o pH do rúmen, nem sobre os conteúdos de: acetato, propionato, isobutirato, butirato, isovalerato e valerato (p>0,05). No entanto, a suplementação de F. suspensa diminuiu a concentração total de ácidos gordurosos voláteis (AGV) em comparação com a dieta controle, independentemente das concentrações de F. suspensa (p<0,05). As cabras alimentadas com 0,25% F. suspensa na dieta tiveram maiores quantidades da enzima fosfatasse alcalina (ALP) do que quando elas foram alimentadas com a dieta controle ou com 0,50% de F. suspensa (p<0,05). Ainda que a alimentação com diferentes níveis de F. suspensa não influenciou os níveis de IgA e IgM no plasma (p>0,05), as cabras que receberam na dieta 0,25 e 0,50% de F. suspensa tiveram uma maior concentração de IgG no plasma comparadas as cabras alimentadas com a dieta controle (p<0,05). Conclusões: a suplementação com 0,25% e 50% diminuiu significativamente a concentração de AGV e incrementou a concentração em plasma de IgG em cabras nativas coreanas comparadas com aquelas que consumiram a dieta controle.

Understanding the adsorptive and photoactivity properties of Ag-graphene oxide nanocomposites.

Nanocomposites of graphene oxide (GO) and silver nanoparticles (AgNPs) were synthetized using a practical photochemical silver functionalization. Their photocatalytic activities were evaluated with two dyes, Rhodamine B and Indigo Carmine, under visible-light irradiation. The prepared nanocomposites were characterized by HRTEM, FESEM, XRD, Raman, FTIR and UV-vis absorption spectroscopy. These nanocomposites present new defect domains of sp(3) type in combination with several graphitic functional groups that act as nucleation sites for anchoring AgNPs, while the sp(2)-sp(3) edge defects domains of GO generate the photoactivity. Furthermore, their photocatalytic performances are governed by their large adsorption capacity, and strong interaction with dye chromophores. A comprehensive photocatalytic way underlying the importance of adsorption is suggested to explain the low visible-light responsive photoactivity of the AgNPs-GO nanocomposites and the possible binding-site saturation. Then, the usage of H2SO4 allows the production of ionic species and helps to confirm the strong adsorption of both dyes. The ability to synthesize AgNPs-GO nanocomposites with extensive adsorptive capacity is certainly of interest for the efficient removal of hazardous materials.

Relationships among sensory analysis, isoflavone and hexanal contents of soymilk powder

The objective of this work was to study the sensory attributes and their relationships with isoflavones and hexanal contents of soymilks made in laboratory and commercial samples. The laboratory soymilk samples showed cooked grain and cotton candy aroma and cooked grain, malty and sweetness flavor (a mild flavor). The commercial samples presented stronger roasted soy, rancid, sesame seeds and fishy aroma and roasted soy, sesame seeds and bitterness flavor, and bitter taste (closed nose) and starchy texture. No differences were noted among laboratory soymilks, denoting inactivation of lipoxygenases enzymes in the soymilks process. There were differences between the samples prepared in laboratory and commercial soymilks, which was due to several factors related to processing techniques. The hexanal average content was positively associated with the cooked grain aroma and isoflavones was positively associated with the cooked grain and cotton candy aroma, cooked grain, malty and sweetness flavor and starchy texture.

O trabalho foi conduzido com o objetivo de descrever os atributos sensoriais de bebidas de extratos de soja em pó produzidos a partir de cultivares desprovida de lipoxigenases, convencional e tipo hortaliça e dois extratos comerciais e correlacioná-los com as quantidades de isoflavonas e n-hexanal nos extratos em pó. Os extratos de soja em pó obtidos em laboratório foram caracterizados pelos atributos: aroma de feijão cozido e de algodão doce; sabor de grão cozido, de malte e gosto doce sugerindo, sabor suave. Os extratos comerciais apresentaram forte aroma de soja tostada, de ranço, de sementes de gergelim e de peixe; sabor de soja tostada, gosto amargo e de sementes de gergelim; gosto amargo (com nariz preso) e consistência de amido. Não foram observadas diferenças sensoriais entre os extratos produzidos em laboratório, indicando a ocorrência da inativação das enzimas lipoxigenases durante o processamento dos produtos. Contudo, diferenças marcantes entre os extratos obtidos em laboratório e os comerciais quanto aos atributos sensoriais estudados foram encontradas provavelmente, devido às técnicas desconhecidas de processamento utilizadas pelos fabricantes dos produtos comerciais. O teor médio de n-hexanal presente nos extratos de soja comerciais e produzidos em laboratório apresentou correlação positiva e significativa com o atributo aroma de grão cozido, já a quantidade de isoflavonas com aroma de grão cozido e de algodão doce, sabor de grão cozido, de malte, gosto doce e consistência de amido.

Phase III randomized trial of sunitinib versus capecitabine in patients with previously treated HER2-negative advanced breast cancer.

This multicenter, randomized, open-label phase III trial (planned enrollment: 700 patients) was conducted to test the hypothesis that single-agent sunitinib improves progression-free survival (PFS) compared with capecitabine as treatment for advanced breast cancer (ABC). Patients with HER2-negative ABC that recurred after anthracycline and taxane therapy were randomized (1:1) to sunitinib 37.5 mg/day or capecitabine 1,250 mg/m(2) (1,000 mg/m(2) in patients >65 years) BID on days 1-14 q3w. The independent data-monitoring committee (DMC) determined during the first interim analysis (238 patients randomized to sunitinib, 244 to capecitabine) that the trial be terminated due to futility in reaching the primary endpoint. No statistical evidence supported the hypothesis that sunitinib improved PFS compared with capecitabine (one-sided P = 0.999). The data indicated that PFS was shorter with sunitinib than capecitabine (median 2.8 vs. 4.2 months, respectively; HR, 1.47; 95% CI, 1.16-1.87; two-sided P = 0.002). Median overall survival (15.3 vs. 24.6 months; HR, 1.17; two-sided P = 0.350) and objective response rates (11 vs. 16%; odds ratio, 0.65; P = 0.109) were numerically inferior with sunitinib versus capecitabine. While no new or unexpected safety findings were reported, sunitinib treatment was associated with higher frequencies and greater severities of many common adverse events (AEs) compared with capecitabine, resulting in more temporary discontinuations due to AEs with sunitinib (66 vs. 51%). The relative dose intensity was lower with sunitinib than capecitabine (73 vs. 95%). Based on these efficacy and safety results, sunitinib should not be used as monotherapy for patients with ABC.

Validity of self-reported weight and height: comparison between immigrant and non-immigrant Mexican Americans in NHANES III.

This study examined whether 1) the immigrant Mexican Americans reported weight and height differently from the non-immigrant Mexican Americans and, 2) self-reporting errors influenced weight classification. A U.S. national sample of 3085 Mexican Americans from the third National Health and Nutrition Examination Survey was used to investigate validity using paired t-tests, multiple linear regressions, sensitivity and specificity. While immigrant Mexican American women reported their weight and height higher than non-immigrant women, no significant differences were found between immigrant and non-immigrant Mexican American men. Sensitivity and specificity of weight classification with self-reported weight and height were not different between immigrant and non-immigrant Mexican Americans; however, underweight Mexican Americans showed the lowest sensitivity. Therefore, self-reported weight and height can be used for the purpose of weight classification with immigrant and non-immigrant Mexican Americans, with an exception of underweight Mexican Americans.

Efeitos da estimulacao cronica do cortex pre-frontal medial esquerdo sobre a aprendizagem e memoria em ratos

Ha relatos de que a hiperfuncao do cortex pre-frontal medial poderia levar a falencia de regioes que recebem suas projecoes, e estao envolvidos nos processos de aprendizagem e memoria. Este estudo avaliou o desempenho de ratos submetidos a estimulacao eletrica repetitiva do cortex pre-frontal medial esquerdo (CPFME) na aquisicao e execucao de tarefas no labirinto radial de 8 bracos. Ratos Wistar com eletrodos implantados no CPFME (B=2 mmA; 0,6 mmL e 2,7 mmV) foram divididos em 3 grupos de 6 animais: CON (sem estimulacao, mas submetidos a mesma manipulacao dos demais), CTE (estimulacoes de intensidade fixa de 50 amperesA) e VAR (estimulacoes de intensidade variavel capazes de induzir comportamentos estereotipados). Foram realizadas sessoes diarias de estimulacao, a tarde, e de avaliacao no labirinto radial, pela manha. Inicialmente os animais foram treinados para a aquisicao da tarefa (aprendizagem). Posteriormente foram submetidos a testes com retardos (interposicoes de intervalos d eretencao na metade da execucao da sessao) de 5 s e 1 h para avaliar o tempo de curso da memoria operacional) Os resultados demonstraram que animais submetidos a estimulacao eletrica repetitiva do CPFME nao apresentaram alteracoes na aprendizagem e na memoria operacional avaliadas pela execucao de testes no labirinto radial.