Sensing danger through the olfactory system: The role of the hypothalamic dorsal premammillary nucleus

The dorsal premammillary nucleus (PMd) has a critical role on the expression of defensive responses to predator odor. Anatomical evidence suggests that the PMd should also modulate memory processing through a projecting branch to the anterior thalamus. By using a pharmacological blockade of the PMd with the NMDA-receptor antagonist 2-amino-5-phosphonopentanoic acid (AP5), we were able to confirm its role in the expression of unconditioned defensive responses, and further revealed that the nucleus is also involved in influencing associative mechanisms linking predatory threats to the related context. We have also tested whether olfactory fear conditioning, using coffee odor as CS, would be useful to model predator odor. Similar to cat odor, shock-paired coffee odor produced robust defensive behavior during exposure to the odor and to the associated context. Shock-paired coffee odor also up-regulated Fos expression in the PMd, and, as with cat odor, we showed that this nucleus is involved in the conditioned defensive responses to the shock-paired coffee odor and the contextual responses to the associated environment. (C) 2008 Elsevier Ltd. All rights reserved.

Absence of Melatonin Induces Night-Time Hepatic Insulin Resistance and Increased Gluconeogenesis Due to Stimulation of Nocturnal Unfolded Protein Response

It is known that the circadian rhythm in hepatic phosphoenolpyruvate carboxykinase expression (a limiting catalytic step of gluconeogenesis) and hepatic glucose production is maintained by both daily oscillation in autonomic inputs to the liver and night feeding behavior. However, increased glycemia and reduced melatonin (Mel) levels have been recently shown to coexist in diabetic patients at the end of the night period. In parallel, pinealectomy (PINX) is known to cause glucose intolerance with increased basal glycemia exclusively at the end of the night. The mechanisms that underlie this metabolic feature are not completely understood. Here, we demonstrate that PINX rats show night-time hepatic insulin resistance characterized by reduced insulin-stimulated RAC-alpha serine/threonine-protein kinase phosphorylation and increased phosphoenolpyruvate carboxykinase expression. In addition, PINX rats display increased conversion of pyruvate into glucose at the end of the night. The regulatory mechanism suggests the participation of unfolded protein response (UPR), because PINX induces night-time increase in activating transcription factor 6 expression and prompts a circadian fashion of immunoglobulin heavy chain-binding protein, activating transcription factor 4, and CCAAT/enhancer-binding protein-homologous protein expression with Zenith values at the dark period. PINX also caused a night-time increase in Tribble 3 and regulatory-associated protein of mammalian target of rapamycin; both were reduced in liver of PINX rats treated with Mel. Treatment of PINX rats with 4-phenyl butyric acid, an inhibitor of UPR, restored night-time hepatic insulin sensitivity and abrogated gluconeogenesis in PINX rats. Altogether, the present data show that a circadian oscillation of UPR occurs in the liver due to the absence of Mel. The nocturnal UPR activation is related with night-time hepatic insulin resistance and increased gluconeogenesis in PINX rats. (Endocrinology 152: 1253-1263, 2011)

Conformational Properties of Angiotensin II and Its Active and Inactive TOAC-Labeled Analogs in the Presence of Micelles. Electron Paramagnetic Resonance, Fluorescence, and Circular Dichroism Studies

The interaction between angiotensin II (AII, DRVYIHPF) and its analogs carrying 2,2,6,6-tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid (TOAC) and detergents-negatively charged sodium dodecyl sulfate (SDS) and zwitterionic N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (HPS)-was examined by means of EPR, CD, and fluorescence. EPR spectra of partially active TOAC(1)-AII and inactive TOAC(3)-AII in aqueous solution indicated fast tumbling, the freedom of motion being greater at the N-terminus. Line broadening occurred upon interaction with micelles. Below SDS critical micelle concentration, broader lines indicated complex formation with tighter molecular packing than in micelles. Small changes in hyperfine splittings evinced TOAC location at the micelle-water interface. The interaction with anionic micelles was more effective than with zwitterionic micelles. Peptide-micelle interaction caused fluorescence increase. The TOAC-promoted intramolecular fluorescence quenching was more, pronounced for TOAC(3)-AII because of the proximity between the nitroxide and Tyr(4). CD spectra showed that although both AII and TOAC(1)-AII presented flexible conformations in water, TOAC(3)-AII displayed conformational restriction because of the TOAC-imposed bend (Schreier et al., Biopolymers 2004, 74, 389). In HPS, conformational changes were observed for the labeled peptides at neutral and basic pH. In SDS, all peptides underwent pH-dependent conformational changes. Although the spectra suggested similar folds for All and TOAC(1)-AII, different conformations were acquired by TOAC(3)-AII. The membrane environment has been hypothesized to shift conformational equilibria so as to stabilize the receptor-bound conformation of ligands. The fact that TOAC(3)-AII is unable to acquire conformations similar to those of native AII and partially active TOAC(1)-AII is probably the explanation for its lack of biological activity. (C) 2009 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 92: 525-537, 2009.

A proposed EPR approach to evaluating agonist binding site of a peptide receptor

Angiotensin II (Ang II) and its transmembrane AT(1) receptor were selected in order to test an innovative strategy that might allow the assessment of the agonist binding site in the receptor molecule. With the use of the 2,2,6,6-tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid (TOAC) paramagnetic probe, a biologically active agonist (TOAC(1)-Ang II), as well as an inactive control (TOAC(4)-Ang II) analogs were mixed in solution with various synthesized AT(1) fragments. Comparative intermolecular interactions, as estimated by analyzing the EPR spectra of solutions, suggested the existence of an agonist binding site containing a sequence composed of portions of the N-terminal (13-17) and the third extracellular loop (266-278) fragments of the AT(1) molecule. Therefore, this combined EPR-TOAC approach shows promise as an alternative for use also in other applications related to specific intermolecular association processes.

Enraizamento de estacas de Caesalpinia echinata Lam. em hidroponia

This study aimed to determine the indole-3-butyric acid (IBA) requirement and proper concentration for Caesalpinia echinata rooting in a hydroponic system for plantlet production. The experiment was conducted in a greenhouse of the 'Faculdade de Ciencias Agrarias e Veterinarias', UNESP, in Jaboticabal, São Paulo state. The vegetal material consisted of 160 stem cuttings that were 11 to 12 cm in length with two pairs of leaves, prepared from young seedlings. The cutting bases were treated by slow immersion in 5% ethanol solution containing 0, 100, 200 and 400 mg L-1 of IBA during 14 hours and by fast immersion in 50% ethanol solution containing 0, 1000, 3000 and 5000 mg L-1 of IBA during 5 seconds. The cuttings were maintained in the hydroponic system for 90 days and, for additional 90 days in plastic bags containing substrate (Plantmax (R)) under intermittent mist. After 90 and 180 days of rooting, the percentage of cuttings that survived, the rooting percentage and the number and length of adventious root were evaluated. Cuttings should be treated with IBA at 100 mg L-1 for 14 hours and placed to root in a hydroponic system.

Propagação de jambeiro vermelho (Syzygium malaccense L.) por estaquia de ramos herbáceos

The consumption of exotic fruits has been showing accentuated increase and the cultivated area is in expansion, generating demand for adequate culturing techniques. The Malay apple (Syzygium malaccense), with probable origin in India, has a fruit widely known and appreciated in the North and Northeasthern Brazilian states. The Malay apple tree is extremely tall and has a long juvenile period when propagated by seed, making its vegetative multiplication is desirable, to anticipate the productive period and decrease its size, and also to obtain uniform orchards. The experiment was conducted at UNESP/FCAV, Jaboticabal Campus, using Malay apple herbaceous cuttings subjected to treatments with indol butyric acid (IBA) (0; 1,000; 3,000 and 5,000 mgL(-1)) and cuttings with and without basal incision. The variables analyzed were percentage of survival and rooting of the cuttings, number and mean length of roots per cutting. The experiment was conducted under CRB on a factorial scheme (4 X 2) with 4 replicates constituted by 10 cuttings each. Data were analyzed by Tukey's mean test at 5% probability. The vegetative propagation by rooting of herbaceous cuttings of the Malay apple is possible, however, both IBA treatments and basal incision have not shown significant effect on the analyzed variables.

Influência do ácido indolbutírico no enraizamento de estacas apicais e basais de caramboleira (Averrhoa carambola L.) sob condições de nebulização intermitente

A produção de mudas de caramboleira é um dos fatores limitantes à expansão comercial da cultura, devido ao tempo que estas levam para serem formadas e iniciarem a produção. Uma boa alternativa pode estar na otimização dos métodos de propagação vegetativa, através de estudos relativos aos processos e fatores envolvidos no enraizamento de estacas. Este trabalho foi desenvolvido com o objetivo de estudar o enraizamento de estacas apicais e basais de caramboleira, tratadas com ácido indolbutírico (IBA), em condições de nebulização intermitente. Estacas apicais e basais de caramboleira foram coletadas de ramos de plantas-matrizes da cultivar B-10 e submetidas à aplicação de cinco concentrações de IBA (0; 1.000; 3.000; 5.000 e 7000 mg.L-1), em imersão por 10 segundos, para avaliar a capacidade de sobrevivência, enraizamento e número médio de raízes/estaca. Posteriormente, as estacas foram colocadas em caixas de madeira contendo vermiculita média como substrato e mantidas em casa de vegetação, sob nebulização intermitente, durante 70 dias. O delineamento experimental utilizado foi em esquema fatorial 2 x 5, com 4 repetições e 10 estacas/parcela. As estacas apicais obtiveram melhores resultados para porcentagem de sobrevivência (49,26%), enraizamento (34,84%) e número médio de raízes (20,51), mostrando-se superiores às basais. O uso de IBA não influenciou em nenhuma das variáveis analisadas.

Estaquia como processo de clonagem do bacuripari (Redhia gardneriana Miers ex Planch e Triana)

No Brasil, existem diversas fruteiras nativas com potencial de exploração comercial, especialmente na Amazônia, local de origem do bacuripari. Neste sentido, realizou-se um trabalho com o objetivo de avaliar a clonagem dessa espécie pelo processo da estaquia, mediante uso de ácido indolil-3-butírico (AIB), em condições de nebulização intermitente. O delineamento experimental foi o inteiramente casualizado, com 5 tratamentos, caracterizados pelas concentrações de AIB (0; 1.000; 3.000; 5.000 e 7.000 mg.L-1), com 4 repetições e 10 estacas por parcela. Foram avaliados na porcentagem de estacas enraizadas, o número médio de folhas e o comprimento de raízes. O uso de AIB não influenciou na porcentagem de enraizamento.

Influência da frequência de suplementação no consumo, na digestibilidade e na fermentação ruminal em novilhos de corte mantidos em pastagem de capim-marandu

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação da época de estaquia e uso de bioregulador no enraizamento de estacas de figueira

O trabalho objetivou avaliar a propagação da figueira com estacas semilenhosas, empregando-se concentrações de ácido indolbutírico, em diferentes épocas de coleta. O material propagativo utilizado foi proveniente de figueiras da cv. Roxo de Valinhos, com 5 anos de idade. As estacas foram retiradas por ocasião da poda hibernal, no final dos meses de agosto, setembro e outubro, as quais foram tratadas com AIB preparado em pó, nas seguintes concentrações: 0 (testemunha); 2.500; 5.000; 7.500 e 10.000 mg kg-1. Posteriormente, foram colocadas para enraizar em bandejas de polipropileno, tendo como substrato vermiculita e mantidas sob nebulização intermitente por 70 dias. Decorrido esse período, avaliaram-se a porcentagem de estacas enraizadas (%), o comprimento da maior raiz (cm) e a massa seca das raízes (g). O delineamento experimental utilizado foi o inteiramente casualizado, em esquema fatorial 3 x 5. Observou-se que estacas provenientes do tratamento-testemunha e da poda de agosto apresentaram baixa porcentagem de enraizamento (20%), menor comprimento e massa seca de raiz, necessitando de tratamento com AIB na concentração de 2.500 mg kg-1, que aumentou significativamente esta porcentagem (90%). Estacas oriundas da poda de setembro e outubro não necessitaram de tratamento com AIB para conseguirem um elevado enraizamento. O mês de setembro foi o mais viável para a coleta de estacas de figueira, pois houve maior porcentagem de enraizamento das estacas (95-100%). As estacas tratadas com 7.500 e 10.000 mg kg-1 de AIB apresentaram as mais baixas porcentagens de enraizamento nos meses de agosto e outubro.

Cultura de embrião e indução de brotos in vitro para micropropagação do pinhão-manso

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Rooting of healthy and CVC-affected 'Valência' sweet orange stem cuttings, through the use of plant regulators

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Enraizamento de estacas de Trichilia catigua A. Juss (catigua) em diferentes estações do ano

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Enraizamento de estacas de brotações oriundas de estacas radiculares de amoreira-preta

This study aimed to evaluate the effect of indolyl-3-butyric acid (IBA) in promoting the rooting of blackberry shoots derived of root cuttings. This work was conducted from June to August 2010, at UNESP, Botucatu, São Paulo state, Brazil, under the randomized block design, with six concentrations of IBA and six replications of 12 shoot cuttings per plot. The treatments consisted of six concentrations of IBA, as a solution: T1= 0 mg L(-1), T2= 250 mg L(-1), T3= 500 mg L(-1), T4= 1000 mg L(-1), T5= 2000 mg L(-1) and T6= 4000 mg L(-1) applied on the basis of shoots cuttings for ten seconds. After 60 days, the percentage of rooting, the root fresh weight, the root dry weight, the length of root and the survival percentage, were evaluated. The concentration of 250 mg L(-1) IBA favored characteristics as fresh and dry weight of roots originating from roots of blackberry. The highest concentrations of IBA inhibited root development and characteristics ??? (please, verify the 'Resumo") of the shoots cuttings.

The effects of extrusion conditions and the addition of volatile compounds and flavour enhancers to corn grits on the retention of the volatile compounds and texture of the extrudates

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)