Atlantic Forest succession over calcareous soil, Parque Estadual Turístico do Alto Ribeira - PETAR, SP

The forest succession after abandonment of slash-and-burn agriculture over calcareous soil in Brazilian Atlantic Forest was assessed. This is one of the world's most threatened Biome, with only 8% remaining. The study area is located over calcareous soil inside the Alto Ribeira Touristic State Park (PETAR), southeast Brazil. The phytossociological survey showed a successional pattern dominated by species of Leguminosae, especially Piptadenia gonoacantha (Mart.) J.F. Macbr. This species occurs in calcareous soils as a substitute of Tibouchina pulchra (Cham.) Cogn. (Melastomataceae) that is the most usual dominant tree species in early succession over acidic soil, which is the most common situation in this Biome. These results are important for a better understanding of Neotropical forest biodiversity and characterize a unique genetic bank in this highly endangered Biome. They are also decisive to support actions regarding rehabilitation of degraded lands and a potential tool for Neotropical forest sustainable management, both inside and around the conservation unit.

Woody community dynamics in two fragments of "cerrado" stricto sensu over a seven-year period (1995-2002), MA, Brazil

This study was conducted in two fragments of "cerrado" stricto sensu in the Gerais de Balsas Colonization Project, located in southern Maranhão, Brazil. The objective was to evaluate the dynamics of the woody plant community, over seven years (1995-2002). Four transects of 160 × 20 m were monitored. All woody plants with a stem diameter > 3 cm, at 0.30 m above ground level, were recorded. In 1995, 983 and 1,177 stems were sampled in fragments 1 and 2, respectively; in 2002, 1057 and 1406 stems were sampled in the same fragments. In 1995, the Shannon diversity indices (H') were 3.07 and 3.33, in fragments 1 and 2, respectively, reaching their maximum value in 2002 of 3.11 and 3.35. The community of fragment 1 showed an increase of 7.5% in density and 4.4% in basal area between 1995 and 2002, while in fragment 2 there was an increment of 19.4% in density and 23.5% in basal area, over the same period. The annual increment in diameter was 0.13 cm year-1 and 0.17 cm year-1 in fragments 1 and 2, respectively. The mortality rate was 2.73% per year in fragment 1 and 4.88% per year in fragment 2, while the recruitment rate was 3.25% per year and 5.86% per year, respectively. The community presented high recruitment and mortality rates compared to the studies conducted in other sites, indicating a community that was highly dynamic in the period studied.

Tacrolimus (FK506) reduces ischemia-induced hippocampal damage in rats: a 7- and 30-day study

The neuroprotective effect of the immunosuppressant agent FK506 was evaluated in rats after brain ischemia induced for 15 min in the 4-vessel occlusion model. In the first experimental series, single doses of 1.0, 3.0 or 6.0 mg FK506/kg were given intravenously (iv) immediately after ischemia. In the second series, FK506 (1.0 mg/kg) was given iv at the beginning of reperfusion, followed by doses applied intraperitoneally (ip) 6, 24, 48, and 72 h post-ischemia. The same protocol was used in the third series except that all 5 doses were given iv. Damage to the hippocampal field CA1 was assessed 7 or 30 days post-ischemia on three different stereotaxic planes along the septotemporal axis of the hippocampus. Ischemia caused marked neurodegeneration on all planes (P<0.001). FK506 failed to provide neuroprotection to CA1 both when applied iv as a single dose of 1.0, 3.0 or 6.0 mg/kg (experiment 1), and after five iv injections of 1.0 mg/kg (experiment 3). In contrast, the repeated administration of FK506 combining iv plus ip administration reduced CA1 cell death on all stereotaxic planes both 7 and 30 days post-ischemia (experiment 2; P<=0.01). Compared to vehicle alone, FK506 reduced rectal temperature in a dose-dependent manner (P<=0.05); however, this effect did not alter normothermia (37ºC). FK506 reduced ischemic brain damage, an effect sustained over time and apparently dependent on repeated doses and on delivery route. The present data extend previous findings on the rat 4-vessel occlusion model, further supporting the possible use of FK506 in the treatment of ischemic brain damage.

Atherosclerosis in aged mice over-expressing the reverse cholesterol transport genes

We determined whether over-expression of one of the three genes involved in reverse cholesterol transport, apolipoprotein (apo) AI, lecithin-cholesterol acyl transferase (LCAT) and cholesteryl ester transfer protein (CETP), or of their combinations influenced the development of diet-induced atherosclerosis. Eight genotypic groups of mice were studied (AI, LCAT, CETP, LCAT/AI, CETP/AI, LCAT/CETP, LCAT/AI/CETP, and non-transgenic) after four months on an atherogenic diet. The extent of atherosclerosis was assessed by morphometric analysis of lipid-stained areas in the aortic roots. The relative influence (R²) of genotype, sex, total cholesterol, and its main sub-fraction levels on atherosclerotic lesion size was determined by multiple linear regression analysis. Whereas apo AI (R² = 0.22, P < 0.001) and CETP (R² = 0.13, P < 0.01) expression reduced lesion size, the LCAT (R² = 0.16, P < 0.005) and LCAT/AI (R² = 0.13, P < 0.003) genotypes had the opposite effect. Logistic regression analysis revealed that the risk of developing atherosclerotic lesions greater than the 50th percentile was 4.3-fold lower for the apo AI transgenic mice than for non-transgenic mice, and was 3.0-fold lower for male than for female mice. These results show that apo AI overexpression decreased the risk of developing large atherosclerotic lesions but was not sufficient to reduce the atherogenic effect of LCAT when both transgenes were co-expressed. On the other hand, CETP expression was sufficient to eliminate the deleterious effect of LCAT and LCAT/AI overexpression. Therefore, increasing each step of the reverse cholesterol transport per se does not necessarily imply protection against atherosclerosis while CETP expression can change specific athero genic scenarios.

Efficacy of the FEV1/FEV6 ratio compared to the FEV1/FVC ratio for the diagnosis of airway obstruction in subjects aged 40 years or over

The present cross-sectional, population-based study was designed to evaluate the performance of the FEV1/FEV6 ratio for the detection of airway-obstructed subjects compared to the FEV1/FVC <0.70 fixed ratio test, as well as the lower limit of normality (LLN) for 1000 subjects ³40 years of age in the metropolitan area of São Paulo, SP, Brazil. After the exclusion of 37 (3.7%) spirometries, a total of 963 pre-bronchodilator (BD) and 918 post-BD curves were constructed. The majority of the post-BD curves (93.1%) were of very good quality and achieved grade A (762 curves) or B (93 curves). The FEV1/FEV6 and FEV1/FVC ratios were highly correlated (r² = 0.92, P < 0.000). Two receiver operator characteristic curves were constructed in order to express the imbalance between the sensitivity and specificity of the FEV1/FEV6 ratio compared to two FEV1/FVC cut-off points for airway obstruction: equal to 70 (area under the curve = 0.98, P < 0.0001) and the LLN (area under the curve = 0.97, P < 0.0001), in the post-BD curves. According to an FEV1/FVC <0.70, the cut-off point for the FEV1/FEV6 ratio with the highest sum for sensitivity and specificity was 0.75. The FEV1/FEV6 ratio can be considered to be a good alternative to the FEV1/FVC ratio for the diagnosis of airway obstruction, both using a fixed cut-off point or below the LLN as reference. The FEV1/FEV6 ratio has the additional advantage of being an easier maneuver for the subjects and for the lung function technicians, providing a higher reproducibility than traditional spirometry maneuvers.

Over-expression of cyclooxygenase-2 in endoscopic biopsies of ectopic gastric mucosa

Ectopic gastric mucosa (EGM) is considered to be a congenital condition. Rare cases of adenocarcinoma have been described. There are no data justifying regular biopsies or follow-up. Cyclooxygenase-2 (COX-2) is a protein involved in gastrointestinal tumor development by inhibiting apoptosis and regulating angiogenesis. The aim of this prospective study was to evaluate COX-2 expression in EGM and compare it with normal tissue and Barrett's esophagus. We evaluated 1327 patients. Biopsies were taken from the inlet patch for histological evaluation and from the gastric antrum to assess Helicobacter pylori infection. Biopsies taken from normal esophageal, gastric antrum and body mucosa and Barrett's esophagus were retrieved from a tissue bank. EGM biopsies were evaluated with respect to type of epithelium, presence of H. pylori, and inflammation. COX-2 was detected by immunohistochemistry using the avidin-biotin complex. EGM islets were found in 14 patients (1.1%). Histological examination revealed fundic type epithelium in 58.3% of cases, H. pylori was present in 50% and chronic inflammation in 66.7%. Expression of COX-2 was negative in normal distal esophagus, normal gastric antrum and normal gastric body specimens (10 each). In contrast, EGM presented over-expression of COX-2 in 41.7% of cases and Barrett's esophagus in 90% of cases (P = 0.04 and 0.03, respectively). COX-2 immunoexpression in EGM was not related to gender, age, epithelium type, presence of inflammation or intestinal metaplasia, H. pylori infection, or any endoscopic finding. Our results demonstrate up-regulation of COX-2 in EGM, suggesting a possible malignant potential of this so-called harmless mucosa.

Participation of the endoplasmic reticulum protein chaperone thio-oxidoreductase in gonadotropin-releasing hormone receptor expression at the plasma membrane

Chaperone members of the protein disulfide isomerase family can catalyze the thiol-disulfide exchange reaction with pairs of cysteines. There are 14 protein disulfide isomerase family members, but the ability to catalyze a thiol disulfide exchange reaction has not been demonstrated for all of them. Human endoplasmic reticulum protein chaperone thio-oxidoreductase (ERp18) shows partial oxidative activity as a protein disulfide isomerase. The aim of the present study was to evaluate the participation of ERp18 in gonadotropin-releasing hormone receptor (GnRHR) expression at the plasma membrane. Cos-7 cells were cultured, plated, and transfected with 25 ng (unless indicated) wild-type human GnRHR (hGnRHR) or mutant GnRHR (Cys14Ala and Cys200Ala) and pcDNA3.1 without insert (empty vector) or ERp18 cDNA (75 ng/well), pre-loaded for 18 h with 1 µCi myo-[2-3H(N)]-inositol in 0.25 mL DMEM and treated for 2 h with buserelin. We observed a decrease in maximal inositol phosphate (IP) production in response to buserelin in the cells co-transfected with hGnRHR, and a decrease from 20 to 75 ng of ERp18 compared with cells co-transfected with hGnRHR and empty vector. The decrease in maximal IP was proportional to the amount of ERp18 DNA over the range examined. Mutants (Cys14Ala and Cys200Ala) that could not form the Cys14-Cys200 bridge essential for plasma membrane routing of the hGnRHR did not modify maximal IP production when they were co-transfected with ERp18. These results suggest that ERp18 has a reduction role on disulfide bonds in wild-type hGnRHR folding.

Lack of tolerance for the anti-dyskinetic effects of 7-nitroindazole, a neuronal nitric oxide synthase inhibitor, in rats

7-Nitroindazole (7-NI) inhibits neuronal nitric oxide synthase in vivo and reduces l-DOPA-induced dyskinesias in a rat model of parkinsonism. The aim of the present study was to determine if the anti-dyskinetic effect of 7-NI was subject to tolerance after repeated treatment and if this drug could interfere with the priming effect of l-DOPA. Adult male Wistar rats (200-250 g) with unilateral depletion of dopamine in the substantia nigra compacta were treated with l-DOPA (30 mg/kg) for 34 days. On the 1st day, 6 rats received ip saline and 6 received ip 7-NI (30 mg/kg) before l-DOPA. From the 2nd to the 26th day, all rats received l-DOPA daily and, from the 27th to the 34th day, they also received 7-NI before l-DOPA. Animals were evaluated before the drug and 1 h after l-DOPA using an abnormal involuntary movement scale and a stepping test. All rats had a similar initial motor deficit. 7-NI decreased abnormal involuntary movement induced by l-DOPA and the effect was maintained during the experiment before 7-NI, median (interquartile interval), day 26: 16.75 (15.88-17.00); day 28: 0.00 (0.00-9.63); day 29: 13.75 (2.25-15.50); day 30: 0.5 (0.00-6.25); day 31: 4.00 (0.00-7.13), and day 34: 0.5 (0.00-14.63), Friedman followed by Wilcoxon test,vs day 26, P < 0.05;. The response to l-DOPA alone was not modified by the use of 7-NI before the first administration of the drug (l-DOPA vs time interaction, F1,10 = 1.5, NS). The data suggest that tolerance to the anti-dyskinetic effects of a neuronal nitric oxide synthase inhibitor does not develop over a short-term period of repeated administration. These observations open a possible new therapeutic approach to motor complications of chronic l-DOPA therapy in patients with Parkinson’s disease.

Characterization and analysis of the outcome of adults with acute myeloid leukemia treated in a Brazilian University hospital over three decades

We evaluated the outcome of 227 patients with acute myeloid leukemia during three decades (period 1 - 1980’s, N = 89; period 2 - 1990’s, N = 73; period 3 - 2000’s, N = 65) at a single institution. Major differences between the three groups included a higher median age, rates of multilineage dysplasia and co-morbidities, and a lower rate of clinical manifestations of advanced leukemia in recent years. The proportion of patients who received induction remission chemotherapy was 66, 75, and 85% for periods 1, 2, and 3, respectively (P = 0.04). The median survival was 40, 77, and 112 days, and the 5-year overall survival was 7, 13, and 22%, respectively (P = 0.01). The median disease-free survival was 266, 278, and 386 days (P = 0.049). Survival expectation for patients with acute myeloid leukemia has substantially improved during this 30-year period, due to a combination of lower tumor burden and a more efficient use of chemotherapy and supportive care.

Cotransfected human chondrocytes: over-expression of IGF-I and SOX9 enhances the synthesis of cartilage matrix components collagen-II and glycosaminoglycans

Damage to cartilage causes a loss of type II collagen (Col-II) and glycosaminoglycans (GAG). To restore the original cartilage architecture, cell factors that stimulate Col-II and GAG production are needed. Insulin-like growth factor I (IGF-I) and transcription factor SOX9are essential for the synthesis of cartilage matrix, chondrocyte proliferation, and phenotype maintenance. We evaluated the combined effect of IGF-I and SOX9 transgene expression on Col-II and GAG production by cultured human articular chondrocytes. Transient transfection and cotransfection were performed using two mammalian expression plasmids (pCMV-SPORT6), one for each transgene. At day 9 post-transfection, the chondrocytes that were over-expressing IGF-I/SOX9 showed 2-fold increased mRNA expression of the Col-II gene, as well as a 57% increase in Col-II protein, whereas type I collagen expression (Col-I) was decreased by 59.3% compared with controls. The production of GAG by these cells increased significantly compared with the controls at day 9 (3.3- vs 1.8-times, an increase of almost 83%). Thus, IGF-I/SOX9 cotransfected chondrocytes may be useful for cell-based articular cartilage therapies.

Sementes de ipê-branco (Tabebuia roseo-alba (Ridl.) Sand. - Bignoniaceae): temperatura e substrato para o teste de germinação

A pesquisa avaliou a temperatura e o substrato para o teste de germinação das sementes de ipê-branco (Tabebuia roseo-alba (Ridl.) Sand.). Em mesa termogradiente, foram avaliados 11 intervalos de temperatura entre 15°C e 35°C, sob oito horas de fotoperíodo diário. Foram analisados também os substratos papel (sobre papel e rolo de papel) e vermiculita (entre vermiculita) sob 30°C e 35°C, em germinadores com oito horas de fotoperíodo diário. As sementes e as plântulas foram avaliadas diariamente, considerando os critérios indicados nas regras para análise de sementes, e calculados a porcentagem e o índice de velocidade de germinação. A temperatura e o substrato interferem na germinação das sementes de ipê-branco; a condição mais favorável para o teste de germinação dessas sementes é 30°C em substrato papel.

Armazenamento de sementes de ipê-branco e ipê-roxo em diferentes embalagens e ambientes

O armazenamento em condições ideais é de fundamental importância para a conservação da qualidade fisiológica de um lote de sementes. Nesse trabalho procurou-se identificar a condição adequada para o armazenamento de sementes de ipê-branco (Tabebuia roseo-alba (Ridl.) Sand.) e de ipê-roxo (Tabebuia impetiginosa (Mart.) Standl.), bem como avaliar métodos para estimar alterações na viabilidade e no vigor dessas sementes. Diferentes grupos de sementes foram acondicionados em saco de polietileno, saco de papel Kraft e lata e armazenados por até 300 dias em condições de laboratório, geladeira e câmara refrigerada. No início do armazenamento e a cada 60 dias, determinaram-se o teor de água, a porcentagem e a velocidade de germinação e a condutividade elétrica da solução de embebição das sementes. A melhor condição para conservação de sementes de T. roseo-alba e T. impetiginosa foi obtida com o acondicionamento em lata e manutenção em geladeira. Sementes de T. impetiginosa também podem ser conservadas embaladas em saco de polietileno, saco de papel ou lata quando estocadas em câmara refrigerada. Alterações no vigor de sementes dessas espécies são primeiramente identificadas pela redução da velocidade de germinação.

Conservação de sementes de ipê-roxo (Tabebuia impetiginosa (Mart. ex DC.) Standl.) em nitrogênio líquido

O ipê-roxo (Tabebuia impetiginosa (Mart. ex DC.) Standl.) é espécie de alto valor econômico, ornamental e medicinal e está na relação das espécies para conservação genética ex situ, no Instituto Florestal de São Paulo. O objetivo deste trabalho foi o de estudar o comportamento fisiológico de sementes de ipê-roxo, conservadas com diferentes teores de água em nitrogênio líquido, por 360 dias. Frutos maduros foram colocados sobre bancada de alvenaria, em interior de laboratório e sob luz indireta, para posterior extração manual das sementes. Primeiramente, foi determinado o grau de umidade do lote e obtida a amostra do tratamento controle, com o maior teor de água a ser estudado (18,3%). Em seguida, as sementes remanescentes foram submetidas à secagem, em secador com circulação de ar a 30ºC, para a obtenção dos demais tratamentos (12,5, 8,4 e 4,2% de água). Amostras de sementes de cada um dos teores de água foram armazenadas em nitrogênio líquido (-196ºC), sendo submetidas às avaliações fisiológicas no início e após 120, 240 e 360 dias de armazenamento. O delineamento experimental foi o inteiramente casualizado com quatro tratamentos (graus de umidade), avaliados separadamente para cada época de armazenamento. A comparação das médias foi realizada pelo teste Tukey, a 5% de probabilidade. As sementes de ipê-roxo podem ser previamente desidratadas a baixo teor de água (4,2%) e, posteriormente, conservadas em nitrogênio líquido por, pelo menos, 360 dias.

Conservação de sementes de ipê-amarelo (Tabebuia chrysotricha (Mart. ex A. DC.) standl.) em função do teor de água das sementes e da temperatura do armazenamento

O ipê-amarelo (Tabebuia chrysotricha (Mart. ex A. DC.) Standl.) é uma espécie arbórea de alto valor ecológico, econômico e paisagístico. Suas sementes apresentam variação significativa na qualidade durante o armazenamento, o que representa dificuldades no desenvolvimento de técnicas de propagação. O objetivo desta pesquisa foi estudar o comportamento fisiológico das sementes de ipê-amarelo durante o armazenamento através de variações no teor de água das sementes e na temperatura do ambiente. Frutos colhidos manualmente de plantas matrizes foram colocados para secagem e posterior extração das sementes. Primeiramente, foi determinado o teor de água inicial do lote e, paralelamente, obtida a porção com o maior teor de água a ser estudado (21,1%). Em seguida, as sementes remanescentes foram submetidas à secagem, em secador com circulação forçada de ar a 28 ± 2ºC, por diferentes períodos, obtendo-se teores de água de 15,9, 13,6, 11,9 e 8,5%. As porções correspondentes aos diferentes teores de água foram armazenadas a -12, 10 e 20ºC. No início e a cada 30 dias até 270 dias de armazenamento, as sementes foram submetidas às avaliações de teor de água, germinação, emergência, velocidade de emergência e comprimento da parte aérea. O mais alto e o mais baixo teor de água estudado (21,1% e 8,5%) são prejudiciais à qualidade fisiológica das sementes mesmo à temperatura extremamente baixa (-12ºC) de armazenamento. A conservação das sementes de ipê-amarelo (Tabebuia chrysotricha) é favorecida quando armazenadas com teor de água em torno de 11,9% nas temperaturas de 10 e -12ºC.

Teste de raios X para avaliação do potencial fisiológico de sementes de ipê-roxo

Em espécies multiplicadas comercialmente via semeadura indireta, como várias essências florestais, a utilização de sementes de alto potencial fisiológico é imprescindível para garantir o desenvolvimento e estabelecimento adequado das plantas no campo. Assim, diversas técnicas têm sido utilizadas para avaliar a integridade das sementes e, dentre elas, o teste de raios X. O objetivo no trabalho foi avaliar o potencial fisiológico das sementes de ipê roxo (Tabebuia heptaphylla), após separação por cor e tamanho, por meio dos testes de raios X e de germinação. As sementes foram classificadas pela cor, utilizando carta de cor de Munsell, e tamanho, utilizando peneira de crivos oblongos, obtendo-se materiais das frações amarelo-claro maiores, amarelo-claro menores, amarelo-escuras maiores, amarelo-escuras menores, mistura de amarelo-claro maiores, amarelo-claro menores e amarelo-escuro maiores e testemunha (sem classificação). Em seguida, as sementes das diferentes frações, foram submetidas a avaliações da integridade física e viabilidade, por meio de teste de raios X e de germinação, respectivamente. Adotou-se delineamento experimental inteiramente casualizado, com seis tratamentos e cinco repetições, comparando-se as médias pelo teste de Scott-Knott (p < 0,05). O teste de raios X é eficiente na avaliação da morfologia interna de sementes de ipê-roxo e sua relação com o potencial fisiológico. A coloração e o tamanho das sementes de ipê-roxo influenciam o potencial fisiológico das mesmas.