Cystamine and cysteamine increase brain levels of BDNF in Huntington disease via HSJ1b and transglutaminase

There is no treatment for the neurodegenerative disorder Huntington disease (HD). Cystamine is a candidate drug; however, the mechanisms by which it operates remain unclear. We show here that cystamine increases levels of the heat shock DnaJ-containing protein 1b (HSJ1b) that are low in HD patients. HSJ1b inhibits polyQ-huntingtin¿induced death of striatal neurons and neuronal dysfunction in Caenorhabditis elegans. This neuroprotective effect involves stimulation of the secretory pathway through formation of clathrin-coated vesicles containing brain-derived neurotrophic factor (BDNF). Cystamine increases BDNF secretion from the Golgi region that is blocked by reducing HSJ1b levels or by overexpressing transglutaminase. We demonstrate that cysteamine, the FDA-approved reduced form of cystamine, is neuroprotective in HD mice by increasing BDNF levels in brain. Finally, cysteamine increases serum levels of BDNF in mouse and primate models of HD. Therefore, cysteamine is a potential treatment for HD, and serum BDNF levels can be used as a biomarker for drug efficacy.

Nedd4-2 modulates renal Na+-Cl- cotransporter via the aldosterone-SGK1-Nedd4-2 pathway.

Regulation of renal Na(+) transport is essential for controlling blood pressure, as well as Na(+) and K(+) homeostasis. Aldosterone stimulates Na(+) reabsorption by the Na(+)-Cl(-) cotransporter (NCC) in the distal convoluted tubule (DCT) and by the epithelial Na(+) channel (ENaC) in the late DCT, connecting tubule, and collecting duct. Aldosterone increases ENaC expression by inhibiting the channel's ubiquitylation and degradation; aldosterone promotes serum-glucocorticoid-regulated kinase SGK1-mediated phosphorylation of the ubiquitin-protein ligase Nedd4-2 on serine 328, which prevents the Nedd4-2/ENaC interaction. It is important to note that aldosterone increases NCC protein expression by an unknown post-translational mechanism. Here, we present evidence that Nedd4-2 coimmunoprecipitated with NCC and stimulated NCC ubiquitylation at the surface of transfected HEK293 cells. In Xenopus laevis oocytes, coexpression of NCC with wild-type Nedd4-2, but not its catalytically inactive mutant, strongly decreased NCC activity and surface expression. SGK1 prevented this inhibition in a kinase-dependent manner. Furthermore, deficiency of Nedd4-2 in the renal tubules of mice and in cultured mDCT(15) cells upregulated NCC. In contrast to ENaC, Nedd4-2-mediated inhibition of NCC did not require the PY-like motif of NCC. Moreover, the mutation of Nedd4-2 at either serine 328 or 222 did not affect SGK1 action, and mutation at both sites enhanced Nedd4-2 activity and abolished SGK1-dependent inhibition. Taken together, these results suggest that aldosterone modulates NCC protein expression via a pathway involving SGK1 and Nedd4-2 and provides an explanation for the well-known aldosterone-induced increase in NCC protein expression.

Influência de nitrogênio e potássio, via fertirrigação, em atributos do solo, níveis críticos foliares e produção do coqueiro-anão

O plantio de coqueiro-anão irrigado, com o objetivo de produzir coco para aproveitamento de água, tem se expandido muito no Brasil. As maiores produtividades, obtidas sob irrigação, influenciam nas quantidades de N e de K requeridas pela planta, o que altera as relações entre os seus teores no solo e na planta. Um experimento com plantas de coqueiro-anão, irrigado por microaspersão, foi conduzido em um Argissolo Amarelo da unidade de paisagem dos tabuleiros costeiros no Nordeste do Brasil. Foi utilizada a matriz Pan Puebla III em blocos ao acaso, com quatro repetições. Os tratamentos de fertirrigação foram dez combinações de N e K, utilizando uréia e cloreto de potássio como fontes dos nutrientes, aplicados semanalmente durante 52 meses. Amostras de solo foram coletadas na zona de influência do microaspersor e amostras de folhas foram coletadas nas posições 9 e 14. A aplicação de uréia reduziu o pH e os teores de Ca2+ e Mg2+ no solo. O maior valor de condutividade elétrica do extrato de saturação do solo foi de 0,65 dS m-1, na profundidade 0_5 cm, e decresceu com a profundidade. Os coeficientes de correlação entre os teores de K na folha e no solo, nas profundidades 0_5, 0_20 e 20_40 cm, foram similares, indicando que a amostragem de solo para fertilidade na cultura do coqueiro-anão fertirrigado pode ser feita na profundidade de 0_20 cm. Maior quantidade de massa do fruto e mais volume do albúmen líquido foram obtidos com a menor dose de N, dado o menor número de frutos por árvore. Os tratamentos de N e K não influenciaram o valor do pH da água do coco, porém alteraram o valor do brix. O teor de K na água do coco aumentou com a quantidade de K aplicada e do teor de K na folha. Os níveis críticos de N e K nas folhas 9 e 14 foram 19,2, 20,5, 12,1 e 9,4 g kg-1, respectivamente. O nível crítico de K no solo pelo Mehlich-1, na profundidade 0_20 cm, foi 45 mg dm-3.

Activation of peroxisome proliferator-activated receptor beta/delta inhibits lipopolysaccharide-induced cytokine production in adipocytes by lowering nuclear factor-kappaB activity via extracellular signal-related kinase 1/2.

OBJECTIVE: Chronic activation of the nuclear factor-kappaB (NF-kappaB) in white adipose tissue leads to increased production of pro-inflammatory cytokines, which are involved in the development of insulin resistance. It is presently unknown whether peroxisome proliferator-activated receptor (PPAR) beta/delta activation prevents inflammation in adipocytes. RESEARCH DESIGN AND METHODS AND RESULTS: First, we examined whether the PPARbeta/delta agonist GW501516 prevents lipopolysaccharide (LPS)-induced cytokine production in differentiated 3T3-L1 adipocytes. Treatment with GW501516 blocked LPS-induced IL-6 expression and secretion by adipocytes and the subsequent activation of the signal transducer and activator of transcription 3 (STAT3)-Suppressor of cytokine signaling 3 (SOCS3) pathway. This effect was associated with the capacity of GW501516 to impede LPS-induced NF-kappaB activation. Second, in in vivo studies, white adipose tissue from Zucker diabetic fatty (ZDF) rats, compared with that of lean rats, showed reduced PPARbeta/delta expression and PPAR DNA-binding activity, which was accompanied by enhanced IL-6 expression and NF-kappaB DNA-binding activity. Furthermore, IL-6 expression and NF-kappaB DNA-binding activity was higher in white adipose tissue from PPARbeta/delta-null mice than in wild-type mice. Because mitogen-activated protein kinase-extracellular signal-related kinase (ERK)1/2 (MEK1/2) is involved in LPS-induced NF-kappaB activation in adipocytes, we explored whether PPARbeta/delta prevented NF-kappaB activation by inhibiting this pathway. Interestingly, GW501516 prevented ERK1/2 phosphorylation by LPS. Furthermore, white adipose tissue from animal showing constitutively increased NF-kappaB activity, such as ZDF rats and PPARbeta/delta-null mice, also showed enhanced phospho-ERK1/2 levels. CONCLUSIONS: These findings indicate that activation of PPARbeta/delta inhibits enhanced cytokine production in adipocytes by preventing NF-kappaB activation via ERK1/2, an effect that may help prevent insulin resistance.

Ciclagem de nutrientes via precipitação pluvial total, interna e escoamento pelo tronco em sistema agroflorestal com Gliricidia sepium

A deposição de nutrientes contidos na água de chuva que passa pela copa das árvores e que escoa pelo tronco até o solo pode constituir uma via importante da ciclagem biogeoquímica em sistemas agroflorestais de baixo uso de insumos externos. Todavia, não há informação disponível sobre esses processos em agroecossistemas da região semi-árida do Nordeste brasileiro. O presente trabalho foi conduzido em um sistema agroflorestal de cultivo em aléias, com gliricídia e milho, em Esperança, PB, e teve como objetivo quantificar: a proporção da água de chuva que escoa através da copa das árvores, ou pelo tronco, ou que é interceptada pela copa; e as entradas de N, P e K contidos na água escoada através da copa ou pelo tronco, bem como na água de chuva em áreas sem árvores. Na área experimental, foram delimitadas quatro parcelas, onde foram instalados coletores a 0,50 m de distância do tronco das árvores de gliricídia e coletores tipo ´colarinho´ acoplados ao redor do tronco destas. Paralelamente, foram instalados coletores em áreas adjacentes, sem árvores, para quantificação da precipitação pluvial total. Da precipitação pluvial total, 67 % escoou através da copa , 0,74 % escoou pelo tronco e 32 % foi interceptada pela copa das árvores. As concentrações de N e P foram similares nas amostras da água escoada através da copa ou pelo tronco, porém estas foram cerca de 300 % maiores do que na água de chuva. A concentração de K na água escoada pelo tronco foi cerca de 100 e 600 % maior do que na água escoada através da copa e na água de chuva, respectivamente. Em média, os aportes de N, P e K ao solo foram de 5, 1 e 24 kg ha-1 na água de chuva; 9, 2 e 62 kg ha-1 na água escoada através da copa; e 0,12, 0,02 e 1 kg ha-1 na água escoada pelo tronco, respectivamente. Os resultados demonstram a importância da adoção de sistemas agroflorestais para a sustentabilidade de sistemas agrícolas de baixo uso de insumos externos, como os da região semi-árida do Nordeste.

Uncoupling phosphate deficiency from its major effects on growth and transcriptome via PHO1 expression in Arabidopsis.

Inorganic phosphate (Pi) is one of the most limiting nutrients for plant growth in both natural and agricultural contexts. Pi-deficiency leads to a strong decrease in shoot growth, and triggers extensive changes at the developmental, biochemical and gene expression levels that are presumably aimed at improving the acquisition of this nutrient and sustaining growth. The Arabidopsis thaliana PHO1 gene has previously been shown to participate in the transport of Pi from roots to shoots, and the null pho1 mutant has all the hallmarks associated with shoot Pi deficiency. We show here that A. thaliana plants with a reduced expression of PHO1 in roots have shoot growth similar to Pi-sufficient plants, despite leaves being strongly Pi deficient. Furthermore, the gene expression profile normally triggered by Pi deficiency is suppressed in plants with low PHO1 expression. At comparable levels of shoot Pi supply, the wild type reduces shoot growth but maintains adequate shoot vacuolar Pi content, whereas the PHO1 underexpressor maintains maximal growth with strongly depleted Pi reserves. Expression of the Oryza sativa (rice) PHO1 ortholog in the pho1 null mutant also leads to plants that maintain normal growth and suppression of the Pi-deficiency response, despite the low shoot Pi. These data show that it is possible to unlink low shoot Pi content with the responses normally associated with Pi deficiency through the modulation of PHO1 expression or activity. These data also show that reduced shoot growth is not a direct consequence of Pi deficiency, but is more likely to be a result of extensive gene expression reprogramming triggered by Pi deficiency.

Perdas de nutrientes via subsuperfície em colunas de solo sob fertilização mineral e orgânica

O uso de dejetos de animais na produção agrícola pode ocasionar a contaminação ou poluição das águas por nutrientes, principalmente com aplicações consecutivas na mesma área. Nesse sentido, o objetivo deste trabalho foi avaliar o efeito da aplicação de dejeto líquido de bovinos e fertilizantes minerais na lixiviação de NH4+, NO3-, P e K. Para essa avaliação, foram coletadas colunas indeformadas de solo, num experimento de campo que estava sendo conduzido por sete anos consecutivos e que continha 12 tratamentos, resultantes da combinação de quatro doses de dejeto líquido de bovinos com três doses de fertilizante mineral. O experimento de campo era conduzido num Latossolo Bruno distrófico em sistema de plantio direto com sorgo, milho, aveia-preta e azevém em rotação. Em laboratório, essas colunas, de acordo com os respectivos tratamentos de campo, foram submetidas à aplicação do dejeto líquido de bovinos (0, 30, 60 e 90 m³ ha-¹, aplicados em dose única) e dos fertilizantes minerais (0, 50 e 100 % da dose recomendada para o milho - 120, 60 e 60 kg ha-1 de N, P2O5 e K2O). Imediatamente após a aplicação de dejeto e fertilizante, a água percolada por coluna foi coletada e analisada em cinco turnos contínuos, totalizando 516 mm - aproximadamente três volumes de poros. Constatou-se aumento nas concentrações de N-NH4+, P e K com as doses de dejeto e aumento de N-NO3- e K com as doses de fertilizante mineral. No entanto, com exceção do N-NO3-, as concentrações de N-NH4+ e P estiveram abaixo do limite máximo permitido pela legislação brasileira, indicando a importância da infiltração de água no solo na redução do potencial poluidor dos dejetos.

Activation of c-Jun in the nuclei of neurons of the CA-1 in thrombin preconditioning occurs via PAR-1.

Recently it has been shown that the c-Jun N-terminal kinase (JNK) plays a role in thrombin preconditioning (TPC) in vivo and in vitro. To investigate further the pathways involved in TPC, we performed an immunohistochemical study in hippocampal slice cultures. Here we show that the major target of JNK, the AP-1 transcription factor c-Jun, is activated by phosphorylation in the nuclei of neurons of the CA1 region by using phospho-specific antibodies against the two JNK phosphorylation sites. The activation is early and transient, peaking at 90 min and not present by 3 hr after low-dose thrombin administration. Treatment of cultures with a synthetic thrombin receptor agonist results in the same c-Jun activation profile and protection against subsequent OGD, both of which are prevented by specific JNK inhibitors, showing that thrombin signals through PAR-1 to JNK. By using an antibody against the Ser 73 phosphorylation site of c-Jun, we identify possible additional TPC substrates.

Old world arenaviruses enter the host cell via the multivesicular body and depend on the endosomal sorting complex required for transport.

The highly pathogenic Old World arenavirus Lassa virus (LASV) and the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) use α-dystroglycan as a cellular receptor and enter the host cell by an unusual endocytotic pathway independent of clathrin, caveolin, dynamin, and actin. Upon internalization, the viruses are delivered to acidified endosomes in a Rab5-independent manner bypassing classical routes of incoming vesicular trafficking. Here we sought to identify cellular factors involved in the unusual and largely unknown entry pathway of LASV and LCMV. Cell entry of LASV and LCMV required microtubular transport to late endosomes, consistent with the low fusion pH of the viral envelope glycoproteins. Productive infection with recombinant LCMV expressing LASV envelope glycoprotein (rLCMV-LASVGP) and LCMV depended on phosphatidyl inositol 3-kinase (PI3K) as well as lysobisphosphatidic acid (LBPA), an unusual phospholipid that is involved in the formation of intraluminal vesicles (ILV) of the multivesicular body (MVB) of the late endosome. We provide evidence for a role of the endosomal sorting complex required for transport (ESCRT) in LASV and LCMV cell entry, in particular the ESCRT components Hrs, Tsg101, Vps22, and Vps24, as well as the ESCRT-associated ATPase Vps4 involved in fission of ILV. Productive infection with rLCMV-LASVGP and LCMV also critically depended on the ESCRT-associated protein Alix, which is implicated in membrane dynamics of the MVB/late endosomes. Our study identifies crucial cellular factors implicated in Old World arenavirus cell entry and indicates that LASV and LCMV invade the host cell passing via the MVB/late endosome. Our data further suggest that the virus-receptor complexes undergo sorting into ILV of the MVB mediated by the ESCRT, possibly using a pathway that may be linked to the cellular trafficking and degradation of the cellular receptor.

El Dr. Lluís Via Boada, una vida fecunda

El Dr. Lluís Via Boada, nació en Vilafranca del Penedes en octubre de 1910, y empezó sus estudios eclesiásticos en el Seminario de Barcelona en setiembre de 1925.

Synaptic Plasticity at Intrathalamic Connections via CaV3.3 T-type Ca2+ Channels and GluN2B-Containing NMDA Receptors.

The T-type Ca(2+) channels encoded by the Ca(V)3 genes are well established electrogenic drivers for burst discharge. Here, using Ca(V)3.3(-/-) mice we found that Ca(V)3.3 channels trigger synaptic plasticity in reticular thalamic neurons. Burst discharge via Ca(V)3.3 channels induced long-term potentiation at thalamoreticular inputs when coactivated with GluN2B-containing NMDA receptors, which are the dominant subtype at these synapses. Notably, oscillatory burst discharge of reticular neurons is typical for sleep-related rhythms, suggesting that sleep contributes to strengthening intrathalamic circuits.

Oxygen tension controls the expression of the monocarboxylate transporter MCT4 in cultured mouse cortical astrocytes via a hypoxia-inducible factor-1α-mediated transcriptional regulation.

The monocarboxylate transporter MCT4 is a high capacity carrier important for lactate release from highly glycolytic cells. In the central nervous system, MCT4 is predominantly expressed by astrocytes. Surprisingly, MCT4 expression in cultured astrocytes is low, suggesting that a physiological characteristic, not met in culture conditions, is necessary. Here we demonstrate that reducing oxygen concentration from 21% to either 1 or 0% restored in a concentration-dependent manner the expression of MCT4 at the mRNA and protein levels in cultured astrocytes. This effect was specific for MCT4 since the expression of MCT1, the other astrocytic monocarboxylate transporter present in vitro, was not altered in such conditions. MCT4 expression was shown to be controlled by the transcription factor hypoxia-inducible factor-1α (HIF-1α) since under low oxygen levels, transfecting astrocyte cultures with a siRNA targeting HIF-1α largely prevented MCT4 induction. Moreover, the prolyl hydroxylase inhibitor dimethyloxalylglycine (DMOG) induced MCT4 expression in astrocytes cultured in presence of 21% oxygen. In parallel, glycolytic activity was enhanced by exposure to 1% oxygen as demonstrated by the increased lactate release, an effect dependent on MCT4 expression. Finally, MCT4 expression was found to be necessary for astrocyte survival when exposed for a prolonged period to 1% oxygen. These data suggest that a major determinant of astrocyte MCT4 expression in vivo is likely the oxygen tension. This could be relevant in areas of high neuronal activity and oxygen consumption, favouring astrocytic lactate supply to neurons. Moreover, it could also play an important role for neuronal recovery after an ischemic episode.

Produtividade e qualidade dos grãos de feijão em função da aplicação de nitrogênio em cobertura e via foliar

A adequada disponibilidade de N durante do ciclo do feijoeiro é fundamental para garantir elevada produtividade e qualidade dos grãos produzidos. Esse nutriente pode ser absorvido pelas raízes e folhas da planta. Contudo, ainda existem dúvidas sobre a eficiência da aplicação via foliar de N no feijoeiro e sobre a influência dessa prática na qualidade dos grãos. Objetivou-se neste trabalho avaliar o efeito da adubação nitrogenada em cobertura e via foliar sobre a produtividade e qualidade dos grãos da cultura do feijão. O experimento foi conduzido durante a safra "da seca", em um Latossolo Vermelho distroférrico, no município de Botucatu-SP. O delineamento experimental utilizado foi o de blocos ao acaso com quatro repetições, num esquema fatorial 3 x 4, constituído por três doses de N (0, 45 e 90 kg ha-1) em cobertura e quatro épocas de aplicação de N via foliar (1 - sem aplicação de N via foliar, 2 - pulverização estádio R5 (pré-floração), 3 - pulverização no estádio R7 (início da formação das vagens) e 4 - pulverizações nos estádios R5 e R7). Em cada aplicação de N via foliar foram utilizados 200 L ha-1 de uma solução com 10 % de ureia. Quando foi realizada a adubação nitrogenada de cobertura, a aplicação de N via foliar, independentemente da época, não alterou os componentes da produção, a produtividade e a qualidade dos grãos do feijoeiro. Na ausência da adubação nitrogenada de cobertura, a aplicação de N via foliar na fase reprodutiva aumentou a massa e o tamanho dos grãos, a produtividade de grãos e o teor de proteínas nos grãos do feijoeiro. A aplicação de N via foliar no estádio R5 foi mais eficiente em aumentar a produtividade de grãos do feijoeiro que a aplicação em R7.