Reguladores de crescimento vegetal na concentração de macronutrientes em videira Itália

Em clima tropical semi-árido, o uso de reguladores vegetais em fruteiras permite o manejo do crescimento vegetativo, tão necessário ao escalonamento da produção e ao aumento da fertilidade das gemas. Estas substâncias influenciam vários fenômenos fisiológicos relacionados com a absorção mineral, como a condutância de membrana e utilização metabólica de íons. O objetivo deste trabalho foi avaliar o crescimento vegetativo, produção de matéria seca dos ramos, concentração e quantidade acumulada de N, P, K, Ca e Mg nos ramos e nos pecíolos e produção de panículas florais em resposta ao uso de reguladores vegetais, em plantas de videira da cultivar Itália. No experimento foram avaliados três ciclos vegetativos e cinco tratamentos (controle, chlormequat - 1500 mg L-1; daminozide - 3000 mg L-1; uniconazole - 30 mg L-1 e cloreto de mepiquat - 300 mg L-1), aplicados nas plantas em pulverizações aos 35 e 70 dias após a poda. Com o chlormequat e o uniconazole ocorreu efeito cumulativo nas plantas, quando aplicados em ciclos sucessivos da cultura, favorecendo o acúmulo de nutrientes e a diferenciação de gemas férteis em comparação com as plantas controle das videiras 'Itália'. Embora o daminozide tenha inibido significativamente o crescimento das plantas, não resultou em maior acúmulo de nutrientes e emissão de panículas florais nas plantas tratadas.

Produção e qualidade do milheto semeado em duas épocas e adubado com nitrogênio

O trabalho foi conduzido na FCAV-UNESP, câmpus de Jaboticabal, com o objetivo de avaliar as características fisiológicas de crescimento, produção de matéria seca (MS) e teores de proteína bruta (PB), fibra em detergente ácido (FDA) e digestibilidade in vitro da matéria seca (DIVMS) de dois genótipos de milheto (Pennisetum americanum) cultivar Comum e CMS 02/Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA), semeados em duas épocas (23/11/94 e 10/3/95) e submetidos a quatro doses de N (0; 75; 150 e 225 kg ha-1). O delineamento experimental adotado foi o de blocos ao acaso, com parcelas subdivididas, e três repetições. Na primeira época de semeadura, a cv. Comum apresentou produção de MS total significativamente superior (6.995 kg ha-1) à do genótipo CMS 02 (6.177 kg ha-1), e a aplicação de 150 kg ha-1 foi a dose mais adequada nesse período. Na segunda época de semeadura, a produção de MS total dos genótipos foi, em média, de 2.799 kg ha-1, e a adubação nitrogenada não revelou efeito significativo. Plantas da primeira época de semeadura apresentaram, nos dois primeiros cortes, alta produção de folhas, cujos teores de PB foram superiores a 20%, e os valores de DIVMS em torno de 70%.

Estudo do composto derivado do óleo de Ricinus communis L. (mamona) sobre a bactéria e biopolímero da fermantação etanólica

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo da resposta imune humoral em mulheres com neoplasia ovariana. -

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB

Uso de paclobutrazol na produção de mudas, no crescimento, produção e qualidade de frutos de tomateiro em ambiente protegido

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Investigação da ação da proteína anexina A1 sobre o processo de angiogênese induzido pelo fator de crescimento do endotélio vascular: modelos experimentais in vivo e in vitro

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

Transcriptional effects of 1,25 dihydroxyvitamin D3 physiological and supra-physiological concentrations in breast cancer organotypic culture

Background Vitamin D transcriptional effects were linked to tumor growth control, however, the hormone targets were determined in cell cultures exposed to supra physiological concentrations of 1,25(OH)2D3 (50-100nM). Our aim was to evaluate the transcriptional effects of 1,25(OH)2D3 in a more physiological model of breast cancer, consisting of fresh tumor slices exposed to 1,25(OH)2D3 at concentrations that can be attained in vivo. Methods Tumor samples from post-menopausal breast cancer patients were sliced and cultured for 24 hours with or without 1,25(OH)2D3 0.5nM or 100nM. Gene expression was analyzed by microarray (SAM paired analysis, FDR≤0.1) or RT-qPCR (p≤0.05, Friedman/Wilcoxon test). Expression of candidate genes was then evaluated in mammary epithelial/breast cancer lineages and cancer associated fibroblasts (CAFs), exposed or not to 1,25(OH)2D3 0.5nM, using RT-qPCR, western blot or immunocytochemistry. Results 1,25(OH)2D3 0.5nM or 100nM effects were evaluated in five tumor samples by microarray and seven and 136 genes, respectively, were up-regulated. There was an enrichment of genes containing transcription factor binding sites for the vitamin D receptor (VDR) in samples exposed to 1,25(OH)2D3 near physiological concentration. Genes up-modulated by both 1,25(OH)2D3 concentrations were CYP24A1, DPP4, CA2, EFTUD1, TKTL1, KCNK3. Expression of candidate genes was subsequently evaluated in another 16 samples by RT-qPCR and up-regulation of CYP24A1, DPP4 and CA2 by 1,25(OH)2D3 was confirmed. To evaluate whether the transcripitonal targets of 1,25(OH)2D3 0.5nM were restricted to the epithelial or stromal compartments, gene expression was examined in HB4A, C5.4, SKBR3, MDA-MB231, MCF-7 lineages and CAFs, using RT-qPCR. In epithelial cells, there was a clear induction of CYP24A1, CA2, CD14 and IL1RL1. In fibroblasts, in addition to CYP24A1 induction, there was a trend towards up-regulation of CA2, IL1RL1, and DPP4. A higher protein expression of CD14 in epithelial cells and CA2 and DPP4 in CAFs exposed to 1,25(OH)2D3 0.5nM was detected. Conclusions In breast cancer specimens a short period of 1,25(OH)2D3 exposure at near physiological concentration modestly activates the hormone transcriptional pathway. Induction of CYP24A1, CA2, DPP4, IL1RL1 expression appears to reflect 1,25(OH)2D3 effects in epithelial as well as stromal cells, however, induction of CD14 expression is likely restricted to the epithelial compartment.

Generierung und Transduktion wachstumsnegativer Signale durch den Contactinhibin-Rezeptor

Generierung und Transduktion wachstumsnegativer Signale durch den Contactinhibin-RezeptorDie kontaktabhängige Wachstumshemmung, Kontaktinhibition, basierend auf der Interaktion von Contactinhibin (Ci) und seinem Rezeptor (CiR), reguliert das Zellwachstum. Ziel dieser Arbeit war die Aufklärung der Signalweiterleitung über den CiR.Der transformierende Wachstumsfaktor TGF-beta führt in den meisten Zelltypen wie die Kontaktinhibition zum Zellzyklusstopp in der G1-Phase. Um mögliche Interaktionen der beiden Signalkaskaden zu untersuchen, wurden humane Keratinozyten (HaCaT) mit einem dominant-negativen TGF-beta-Rezeptor Typ II stabil transfiziert. Das Ausschalten des TGF-beta-Signalweges resultierte in einer erhöhten Sättigungsdichte und Aufhebung der Kontaktinhibition. Die durch Kontaktinhibition induzierte Zunahme der Expression der TGF-beta-mRNA bestätigte die mögliche Interaktion der beiden Signalwege.In einem weiteren Ansatz sollte die Proteinkinase C (PKC) als möglicher Second Messenger der Kontaktinhibition untersucht werden. Die Herunterregulierung der PKC-Isoformen alpha, delta, epsilon und mu nach Langzeit-Behandlung mit 12-O-Tetradecanoylphorbol-13-acetat führte in humanen Fibroblasten (FH109) zu einer Reduktion der Kontaktinhibition. Nur durch Inkubation mit dem spezifischen Inhibitor der delta-Isoform Rottlerin konnte die Kontaktinhibition vollständig aufgehoben werden. Eine Beteiligung der PKC-delta in die Kontaktinhibition wurde dadurch bestätigt, daß sowohl die Stärke ihrer Proteinexpression als auch ihre intrazelluläre Verteilung über Zell-Zellkontakte reguliert wurde. Außerdem führte die transiente Transfektion von Maus-Fibroblasten, NIH3T3, mit einer dominant-negativen Mutante der PKC-delta zu einem transformierten Phänotyp.

Risposta Immunitaria Epatite B specifica in pazienti affetti da Epatocarcinoma

Hepatitis B x protein (HBx) is a non structural, multifunctional protein of hepatitis B virus (HBV) that modulates a variety of host processes.Due to its transcriptional activity,able to alter the expression of growth-control genes,it has been implicated in hepatocarcinogenesis.Increased expression of HBx has been reported on the liver tissue samples of hepatocellular carcinoma (HCC),and a specific anti-HBx immune response can be detected in the peripheral blood of patients with chronic HBV.However,its role and entity has not been yet clarified.Thus,we performed a cross-sectional analysis of anti-HBx specific T cell response in HBV-infected patients in different stage of disease.A total of 70 HBV-infected subjects were evaluated:15 affected by chronic hepatitis (CH-median age 45 yrs),14 by cirrhosis (median age 55 yrs),11 with dysplastic nodules (median age 64 yrs),15 with HCC (median age 60 yrs),15 with IC(median age 53 yrs).All patients were infected by virus genotype D with different levels of HBV viremia and most of them (91%) were HBeAb positive.The HBx-specific T cell response was evaluated by anti-Interferon (IFN)-gamma Elispot assay after in vitro stimulation of peripheral blood mononuclear cells,using 20 overlapping synthetic peptides covering all HBx protein sequence.HBx-specific IFN-gamma-secreting T cells were found in 6 out of 15 patients with chronic hepatitis (40%), 3 out of 14 cirrhosis (21%), in 5 out of 11 cirrhosis with macronodules (54%), and in 10 out of 15 HCC patients (67%). The number of responding patients resulted significantly higher in HCC than IC (p=0.02) and cirrhosis (p=0.02). Central specific region of the protein x was preferentially recognize,between 86-88 peptides. HBx response does not correlate with clinical feature disease(AFP,MELD).The HBx specific T-cell response seems to increase accordingly to progression of the disease, being increased in subjects with dysplastic or neoplastic lesions and can represent an additional tool to monitor the patients at high risk to develop HCC

Adjuvant or radical fractionated stereotactic radiotherapy for patients with pituitary functional and nonfunctional macroadenoma

Purpose To evaluate the efficacy and toxicity of stereotactic fractionated radiotherapy (SFRT) for patients with pituitary macroadenoma (PMA). Methods and Materials Between March 2000 and March 2009, 27 patients (male to female ratio, 1.25) with PMA underwent SFRT (median dose, 50.4 Gy). Mean age of the patients was 56.5 years (range, 20.3 - 77.4). In all but one patient, SFRT was administered for salvage treatment after surgical resection (transphenoidal resection in 23, transphenoidal resection followed by craniotomy in 2 and multiple transphenoidal resections in another patient). In 10 (37%) patients, the PMAs were functional (3 ACTH-secreting, 3 prolactinomas, 2 growth hormone-secreting and 2 multiple hormone-secretion). Three (11.1%) and 9 (33.3%) patients had PMA abutting and compressing the optic chiasm, respectively. Mean tumor volume was 2.9 ± 4.6 cm3. Eighteen (66.7%) patients had hypopituitarism prior to SFRT. The mean follow-up period after SFRT was 72.4 ± 37.2 months. Results Tumor size decreased for 6 (22.2%) patients and remained unchanged for 19 (70.4%) other patients. Two (7.4%) patients had tumor growth inside the prescribed treatment volume. The estimated 5-year tumor growth control was 95.5% after SFRT. Biochemical remission occurred in 3 (30%) patients with functional PMA. Two patients with normal anterior pituitary function before SFRT developed new deficits 25 and 65 months after treatment. The 5-year survival without new anterior pituitary deficit was thus 95.8%. Five patients with visual field defect had improved visual function and 1 patient with no visual defect prior to SFRT, but an optic chiasm abutting tumor, had a decline in visual function. The estimated 5-year vision and pituitary function preservation rates were 93.2% and 95.8%, respectively. Conclusions SFRT is a safe and effective treatment for patients with PMA, although longer follow-up is needed to evaluate long-term outcomes. In this study, approximately 1 patient with visual field defect out of two had an improved visual function.

Gene expression in cortex and hippocampus during acute pneumococcal meningitis

BACKGROUND: Pneumococcal meningitis is associated with high mortality (approximately 30%) and morbidity. Up to 50% of survivors are affected by neurological sequelae due to a wide spectrum of brain injury mainly affecting the cortex and hippocampus. Despite this significant disease burden, the genetic program that regulates the host response leading to brain damage as a consequence of bacterial meningitis is largely unknown.We used an infant rat model of pneumococcal meningitis to assess gene expression profiles in cortex and hippocampus at 22 and 44 hours after infection and in controls at 22 h after mock-infection with saline. To analyze the biological significance of the data generated by Affymetrix DNA microarrays, a bioinformatics pipeline was used combining (i) a literature-profiling algorithm to cluster genes based on the vocabulary of abstracts indexed in MEDLINE (NCBI) and (ii) the self-organizing map (SOM), a clustering technique based on covariance in gene expression kinetics. RESULTS: Among 598 genes differentially regulated (change factor > or = 1.5; p < or = 0.05), 77% were automatically assigned to one of 11 functional groups with 94% accuracy. SOM disclosed six patterns of expression kinetics. Genes associated with growth control/neuroplasticity, signal transduction, cell death/survival, cytoskeleton, and immunity were generally upregulated. In contrast, genes related to neurotransmission and lipid metabolism were transiently downregulated on the whole. The majority of the genes associated with ionic homeostasis, neurotransmission, signal transduction and lipid metabolism were differentially regulated specifically in the hippocampus. Of the cell death/survival genes found to be continuously upregulated only in hippocampus, the majority are pro-apoptotic, while those continuously upregulated only in cortex are anti-apoptotic. CONCLUSION: Temporal and spatial analysis of gene expression in experimental pneumococcal meningitis identified potential targets for therapy.

Nitric oxide is protective in listeric meningoencephalitis of rats

The bacterium Listeria monocytogenes causes meningoencephalitis in humans. In rodents, listeriosis is associated with granulomatous lesions in the liver and the spleen, but not with meningoencephalitis. Here, infant rats were infected intracisternally to generate experimental listeric meningoencephalitis. Dose-dependent effects of intracisternal inoculation with L. monocytogenes on survival and activity were noted; 10(4) L. monocytogenes organisms induced a self-limiting brain infection. Bacteria invaded the basal meninges, chorioid plexus and ependyme, spread to subependymal tissue and hippocampus, and disappeared by day 7. This was paralleled by recruitment and subsequent disappearance of macrophages expressing inducible nitric oxide synthase (iNOS) and nitrotyrosine accumulation, an indication of nitric oxide (NO.) production. Treatment with the spin-trapping agent alpha-phenyl-tert-butyl nitrone (PBN) dramatically increased mortality and led to bacterial numbers in the brain 2 orders of magnitude higher than in control animals. Treatment with the selective iNOS inhibitor L-N(6)-(1-iminoethyl)-lysine (L-NIL) increased mortality to a similar extent and led to 1 order of magnitude higher bacterial counts in the brain, compared with controls. The numbers of bacteria that spread to the spleen and liver did not significantly differ among L-NIL-treated, PBN-treated, and control animals. Thus, the infant rat brain is able to mobilize powerful antilisterial mechanisms, and both reactive oxygen and NO. contribute to Listeria growth control.

Culture of ovine bone marrow-derived macrophages and evidence for serum factors distinct from M-CSF contributing to their propagation in vitro.

An in vitro system allowing the culture of ovine bone marrow-derived macrophages (BMMs) is described. Bone marrow (BM) cells from the sternum of 4- to 9-month-old sheep were cultured in liquid suspension in hydrophobic bags with medium containing 20% autologous serum and 20% fetal calf serum (FCS). Cells with macrophage characteristics were positively selected and increased four- to five-fold between day (d) 0 and d18. Granulocytes and cells of lymphoid appearance including progenitor cells were negatively selected and were diminished 50-fold during this 18-d culture. The addition of macrophage colony-stimulating factor (M-CSF)-containing supernatants to liquid cultures did not significantly improve the yield of BMM in 18-d cultures. In contrast, cell survival at d6 and macrophage cell yield at d18 depended on the concentration and source of serum in the culture medium. FCS and 1:1 mixtures of FCS and autologous serum were superior to autologous serum alone. Analysis of growth requirements of ovine BMMs suggested that they are under more complex growth control than their murine counterparts. In an [3H]thymidine incorporation assay with BM cells collected at different times of culture, d3 or d4 BM cells responded to human recombinant M-CSF, human recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF), bovine GM-CSF, murine M-CSF or murine M-CSF-containing supernatants, and bovine interleukin 1 beta (IL-1 beta) in decreasing order of magnitude. Likewise, pure murine BMM populations harvested at d6 responded to homologous GM-CSF, IL-3, and human or murine M-CSF. FCS did not stimulate the proliferation of murine BMMs (d6) and of ovine BM cells (d3 or d4). In contrast, ovine BM cells harvested at d12 responded to FCS by proliferation in a dose-dependent manner but failed to proliferate in the presence of human or murine M-CSF or M-CSF-containing supernatants of mouse and sheep fibroblasts containing mouse macrophage growth-promoting activity. Likewise, various cytokine-containing supernatants and recombinant cytokines (murine IL-3, murine and human GM-CSF, murine and bovine IL-1 beta) did not promote proliferation of ovine d12 BM cells to an extent greater than that achieved with 15% FCS alone. Thus, ovine BMM proliferation is under the control of at least two factors acting in sequence, M-CSF and an unidentified factor contained in FCS. The ovine BMM culture system may provide a model for the analysis of myelomonocytopoiesis in vitro.

The H19/let-7 double-negative feedback loop contributes to glucose metabolism in muscle cells

The H19 lncRNA has been implicated in development and growth control and is associated with human genetic disorders and cancer. Acting as a molecular sponge, H19 inhibits microRNA (miRNA) let-7. Here we report that H19 is significantly decreased in muscle of human subjects with type-2 diabetes and insulin resistant rodents. This decrease leads to increased bioavailability of let-7, causing diminished expression of let-7 targets, which is recapitulated in vitro where H19 depletion results in impaired insulin signaling and decreased glucose uptake. Furthermore, acute hyperinsulinemia downregulates H19, a phenomenon that occurs through PI3K/AKT-dependent phosphorylation of the miRNA processing factor KSRP, which promotes biogenesis of let-7 and its mediated H19 destabilization. Our results reveal a previously undescribed double-negative feedback loop between sponge lncRNA and target miRNA that contributes to glucose regulation in muscle cells.

Interplay between receptor tyrosine kinases and hypoxia signaling in cancer.

Deregulated signaling via receptor tyrosine kinase (RTK) pathways is prevalent in numerous types of human cancers and is commonly correlated with worst prognosis, resistance to various treatment modalities and increased mortality. Likewise, hypoxic tumors are often manifested by aggressive mode of growth and progression following an adaptive genetic reprogramming with consequent transcriptional activation of genes encoding proteins, which support tumor survival under low oxygen-related conditions. Consequently, both the hypoxia-inducible factor (HIF) system, which is the major mediator of hypoxia-related signaling, and numerous RTK systems are considered critical molecular targets in current cancer therapy. It is now evident that there is an intricate molecular crosstalk between RTKs and hypoxia-related signaling in the sense that hypoxia can activate expression of particular RTKs and/or their corresponding ligands, while some RTK systems have been shown to trigger activation of the HIF machinery. Moreover, signaling regulation of some RTK systems under hypoxic conditions has also been documented to take place in a HIF-independent manner. With this review we aim at overviewing the most current observations on that topic and highlight the importance of the potential co-drugging the HIF system along with particular relevant RTKs for better tumor growth control.