The total synthesis of (-)-cryptocaryol A.

A stereoselective total synthesis of (-)-cryptocaryol A (1) is described. Key features of the 17-step route include the use of three boron-mediated aldol reaction-reduction sequences to control all stereocenters and an Ando modification of the Horner-Wadsworth-Emmons olefination that permitted the installation of the Z double bond of the α-pyrone ring.

Effects of immune system status on long-term results of cardiac resynchronization therapy.

AIM: Cardiac resynchronization therapy (CRT) is an effective therapy for patients with reduced systolic function and enlarged QRS. Recently, some Authors have demonstrated that the presence of positive antinuclear antibodies (ANAs) may play a role in the development of heart failure in a population of patients implanted with PM. METHODS: We investigated the effect of positive ANAs in 90 patients (mean age 71±8 years) implanted with a CRT device in our Centre between May 2010 and June 2013. To assess for immunologic contribution to CRT outcome, patients were divided into positive and negative ANAs (ANA +, ANA -), considering as positive patients with an ANAs dilution > 1:80. The primary endpoint was constituted by a combined endpoint of death or first hospitalization for heart failure; secondary endpoints were constituted by: 1) incidence of first hospitalization for heart failure; and 2) total cause mortality. RESULTS: After a mean follow-up of 1200 days, primary endpoint occurred in 11 patients (30%) of ANA+ group and in 8 patients (15.1%) of ANA-group. The significant difference is due to difference in heart failure events (27% vs. 11.3%, P<0.05), whilst difference in total mortality did not reach statistical significance (10.8% vs. 3.8%). CONCLUSION: Immune status seems to play a role in patients with congestive heart failure. If this immunological alteration is a determinant or a consequence of heart failure remains unclear.

Manejo de biorregulador nos componentes de produção e desempenho das plantas de soja / Bioregulators management in parts of production and performance of soybean plants

A grande relevância no posicionamento adequado de novas tecnologias na soja, o objetivo do presente trabalho foi avaliar o desempenho das plantas e os componentes de produção, em resposta à aplicação de biorregulador na cultura da soja. Para tanto, sementes de soja da cultivar BRS 246 RR foram semeadas no mês de outubro dos anos agrícolas de 2007/2008 e 2008/2009, no delineamento experimental em blocos completos com os tratamentos casualizados, com quatro repetições. Os tratamentos, arranjados em esquema fatorial, foram compostos pela combinação do tratamento de sementes com o biorregulador (sem e com 0,500 L 100 kg-1 de sementes) e cinco doses do produto (0; 0,125; 0,250; 0,375 e 0,500 L ha-1) aplicadas via foliar, em dois estádios de desenvolvimento da cultura (V5 ou R3). O biorregulador é composto por três reguladores vegetais nas seguintes concentrações: 0,005% do ácido indolbutírico; 0,009% de cinetina e 0,005% de ácido giberélico. Foram avaliadas características agronômicas e componentes de produção da cultura. O uso do biorregulador influenciou os componentes de produção da cultura, gerando aumentos no número de vagens que foram correspondidos por incremento na produtividade até uma dose máxima, não superior a 340 mL do produto.

The great relevance in the proper positioning of new technologies in soybean, the aim of this study was to evaluate the performance of plants and yield components in response to the application of bioregulators in soybean. For this, seeds of the cultivar BRS 246 RR was planted in October of the crop years 2007/2008 and 2008/2009, the complete block experimental design with treatments design with four replications. The treatments were arranged in factorial, consisted of the combination of seed treatment with bioregulators (with and without 0.500 L-1 100 kg of seeds) and five doses (0, 0.125, 0.250, 0.375 and 0.500 L ha-1) applied to leaves at two developmental stages of culture (V5 or R3). The bioregulators consists of three plant growth regulators in the following concentrations: 0.005% of the IBA, 0.009% 0.005% kinetin and gibberellic acid. The agronomic characteristics and yield components of culture. Use of bioregulators influenced the yield of the crop, causing increases in the number of pods that were matched by increased productivity up to a maximum dose not exceeding 340 mL of product.

Proposta de modelagem não-linear do desempenho germinativo de sementes de milho híbrido / Nonlinear proposal modeling of seed germination performance of hybrid corn seeds

Este trabalho foi desenvolvido com o objetivo de ajustar um modelo de regressão não-linear para estimar o desempenho germinativo de sementes de três lotes comerciais de milho híbrido OC 705, submetidas ao teste de envelhecimento acelerado, à temperatura de 43ºC. Utilizou-se o modelo logístico y(t)=C/(1+exp(B(t-M))) para ajustar os dados. As medidas de viés de Box e de curvaturas paramétrica e intrínseca foram usadas para o diagnóstico do modelo. O modelo proposto ajusta-se adequadamente aos dados de percentuais germinativos de sementes de milho híbrido OC 705, para todos os lotes. Com base nas estimativas dos parâmetros e qualidade de ajuste, o lote três foi identificado como sendo o melhor para comercialização, por apresentar menor redução no poder germinativo ao longo do tempo.

This research was developed with the goal to adjust a nonlinear regression model to estimate seed germination performance of three commercial seed lots of hybrid maize OC 705, subjected to accelerated aging test at a temperature of 43oC. The logistic model y(t)=C/(1+exp (B (t-M))) was used to fit the data. Measures of bias Box and parametric intrinsic bends were used for the diagnosis of the model. The proposed model fits properly to seed germination percentage data of hybrid corn seeds for OC 705, for all seed lots. Based on estimates of the parameters and fit quality, the seed lot three was identified as being the best for marketing, by presenting less reduction in seed germination power over time.

Qualidade fisiológica das sementes de soja rr em resposta ao uso de diferentes tratamentos contendo glyphosate em aplicação sequencial / Physiological quality of rr soybean seeds in response to the use of different treatments with sequential glyphosate application

O objetivo do presente trabalho foi avaliar a qualidade fisiológica das sementes da soja RR, sob efeito do manejo com glyphosate em pós-emergência, aplicado em misturas ou isolado, de forma sequencial. O trabalho foi constituído de três experimentos com distintos tratamentos, instalados no ano agrícola 2006/2007, no município de Floraí, PR. O delineamento experimental utilizado foi o de blocos ao acaso, com doze tratamentos e quatro repetições, para cada experimento. A qualidade fisiológica das sementes foi avaliada por meio dos testes de germinação e primeira contagem do teste de germinação. Os resultados demonstraram que a aplicação sequencial de glyphosate isolado e em altas doses influenciou mais a qualidade das sementes de soja RR, do que a aplicação sequencial em mistura com outros herbicidas, como o lactofen e o chlorimuron-ethyl. Não foi constatado efeito negativo sobre a qualidade fisiológicas das sementes, quando das misturas de glyposate com estes herbicidas em pós-emergência.

O objetivo do presente trabalho foi avaliar a qualidade fisiológica das sementes da soja RR, sob efeito do manejo com glyphosate em pós-emergência, aplicado em misturas ou isolado, de forma sequencial. O trabalho foi constituído de três experimentos com distintos tratamentos, instalados no ano agrícola 2006/2007, no município de Floraí, PR. O delineamento experimental utilizado foi o de blocos ao acaso, com doze tratamentos e quatro repetições, para cada experimento. A qualidade fisiológica das sementes foi avaliada por meio dos testes de germinação e primeira contagem do teste de germinação. Os resultados demonstraram que a aplicação sequencial de glyphosate isolado e em altas doses influenciou mais a qualidade das sementes de soja RR, do que a aplicação sequencial em mistura com outros herbicidas, como o lactofen e o chlorimuron-ethyl. Não foi constatado efeito negativo sobre a qualidade fisiológicas das sementes, quando das misturas de glyposate com estes herbicidas em pós-emergência

Efeito de anos e épocas de semeadura sobre o desempenho agronômico e rendimento de cultivares de trigo em Palotina, PR / Effect of sowing dates and years on agronomic performance and yield of wheat cultivars in Palotina, state of Parana, Brazil

O objetivo do trabalho foi avaliar o desempenho agronômico e rendimento de grãos de cultivares de trigo em função de anos e épocas de semeadura. Os experimentos de campo foram instalados na Fazenda Experimental da Cooperativa Central de Pesquisa Agrícola (Coodetec), em Palotina, Estado do PR, Brasil, durante os anos agrícolas de 2006 e 2007. O delineamento utilizado foi em blocos completos com os tratamentos ao acaso, com quatro repetições. Foram avaliadas seis cultivares de trigo (CD 104, CD 105, CD 108, CD 110, CD 111 e CD 114), e cinco épocas de semeadura (22/03, 07/04, 20/04, 04/05 e 18/05). Realizou-se a análise estatística conjunta para anos, cultivares e épocas de semeadura, em todas as combinações. As características avaliadas foram: ciclo até o espigamento e até a maturação, e rendimento de grãos. A maturação foi a característica mais afetada durante os dois anos avaliados. De maneira geral, em ambos os anos, o maior rendimento de grãos, para as seis cultivares de trigo, foi obtido em semeaduras realizadas no mês de abril.

The objective of this work was to evaluate the agronomic performance and yield of wheat cultivars in response to different sowing dates and years. Field experiments were carried out on the Experimental Farm of the Central Cooperative Unit in Agricultural Research (Coodetec) in Palotina, state of Paraná, Brazil, during the agricultural years of 2006 and 2007. There was used the randomized block design with four replications. There were evaluated six wheat cultivars (CD 104, CD 105, CD 108, CD 110, CD 111 and CD 114), and five sowing dates (03/22, 04/07, 04/20, 05/04 and 05/18). The statistical analysis was carried out jointly for the years, cultivars, and sowing dates, in all combinations. Days until both emergency to flowering and emergency to maturation, and yield grain, were evaluated. Maturation was the characteristic more affected during the two years of study. In general, for both years, the highest grain yield, for the six wheat cultivars, was obtained in April sowing dates.

Desempenho agronômico de três cultivares de soja em diferentes épocas de semeadura em duas safras / Agronomic performance of three soybean cultivars at different sowing times in two agriculture years

O cultivo da soja, com cultivares precoces, em semeadura antecipada, é uma prática comum no oeste do Estado do Paraná, visando à safrinha do milho, no entanto, pouca pesquisa é relatada sobre esse fato. Portanto, neste trabalho,objetivou-se avaliar a influência da antecipação da semeadura da soja, no desempenho agronômico de três cultivares precoces de soja, semeadas em diferentes épocas, em dois anos agrícolas. Para tanto, foram instalados ensaios de competição na unidade da COODETEC, em Palotina - Paraná. O delineamento utilizado foi o em blocos casualizados, com quatro repetições, conduzido em dois anos agrícolas (2003/2004 e 2004/2005), com três cultivares precoces de soja (CD 202, CD 215 e CD 216), semeadas em cinco épocas (15/09, 30/09, 15/10, 30/10 e 15/11). As variáveis avaliadas foram: altura de inserção de primeira vagem, altura de planta, número de vagens e produtividade. Os dados coletados foram submetidos à análise de variância conjunta e realizados os desdobramentos das interações, quando necessário. Constatou-se que: a época preferencial de semeadura foi desfavorável à produção de grãos, especialmente no segundo ano agrícola; a maior produtividade de sementes para as três cultivares de soja foi obtida na semeadura realizada no mês de outubro; a antecipação na semeadura pode ser uma alternativa viável para a região oeste do Estado do Paraná.

Growing soybean by using early maturing soybean cultivars in anticipated sowing has been a common practice in the west of Paraná State aiming at the safrinha corn (fall cropping), however, there has not been enough research reported about this fact. Therefore, the objective of this study was to evaluate the influence of sowing anticipation of three early maturing soybean cultivars during two agriculture years. For this purpose, competition experiments were installed at COODETEC, in Palotina, Paraná State. A randomized complete block design with four replications was carried out within two agriculture years (2003/2004 and 2004/2005) by using three early maturing soybean cultivars (CD 202, CD 215, and CD 216) sowed on five different days (09/15, 09/30, 10/15, 10/30, and 11/15). Data were collected and submitted to variance analysis and the interaction design was accomplished whenever necessary. It was verified that the preferable period for sowing was unfavorable to the seed production, especially in the second agriculture year. The highest seed yield for the three soybean cultivars was obtained from the sowing accomplished in October. Potentially, sowing anticipation might be a viable alternative to the west region of Paraná State.

Influência do estresse hídrico sobre o desempenho fisiológico de sementes de híbridos simples de milho-pipoca / Influence of water stress on the physiological performance of seeds of simple hybrids of popcorn

Objetivou-se, neste trabalho, avaliar o efeito do estresse hídrico induzido por polietileno glicol, na qualidade fisiológica das sementes de catorze híbridos simples de milho-pipoca. Para tanto, foi conduzido um experimento em que as sementes foram colocadas para germinar em substrato de papel-toalha, umedecido com soluções de polietileno glicol (PEG 6000), nos níveis de potencial osmótico de zero (controle) e -0,3 MPa. A qualidade fisiológica das sementes foi avaliada por meio do teste de germinação (primeira contagem e contagem final), do comprimento da raiz primária e da parte aérea das plântulas e da biomassa seca das plântulas. A diminuição do potencial hídrico do substrato é prejudicial à germinação das sementes e, principalmente, ao crescimento das plântulas de milho-pipoca. Os decréscimos nas variáveis analisadas dependeram dos híbridos avaliados. Os híbridos G e K foram os que se mostraram mais tolerantes ao estresse hídrico, enquanto que o híbrido M foi mais susceptível à seca. A tolerância ao estresse hídrico independe da qualidade inicial das sementes de milho-pipoca.

The aim in this work was to evaluate the effect of the water stress induced by polyethylene glycol, in the physiological quality of seeds of fourteen hybrids of popcorn. For this, an experiment was carried out at laboratory in which the seeds were put to germinate in substratum of paper-towel, soaked with solutions of polyethylene glycol (PEG 6000) in the levels of osmotic potential of zero (control) and -0.3 MPa. The physiological quality of the seeds was evaluated through the germination test (first counting and final counting), length of the primary root and aerial part of the seedlings and dry biomass of seedlings. The decrease of the water potential of the substratum is harmful to the germination, mainly to the growth of seedlings of popcorn. The decreases in the analyzed variables depended on the appraised hybrid. The hybrids G and K were the ones that showed to be more tolerant to the water stress, while the one of the hybrid M was more susceptible to the drought. The tolerance to water stress does not depend on the initial quality of the popcorn seeds.

Componentes genéticos de médias e depressão por endogamia em populacões de milho-pipoca / Average genetic components and inbreeding depression in popcorn populations

Dez populacões de milho-pipoca (PR 038, PR 079, RR 046, SC 016, PR 017, BRS Angela, SC 002, PR 009, PR 023 e SE 013) foram avaliadas com os objetivos de obter estimativas de depressão por endogamia e componentes genéticos de média. O delineamento experimental utilizado foi o de blocos ao acaso em esquema de parcelas subdivididas, nos quais os tratamentos primários foram os níveis de endogamia (S1 e S0) e os tratamentos secundários as populacões, com três repeticões. O ensaio foi conduzido nas cidades de Maringá e Iguatemi-PR, no ano agrícola de 2001/2002. Foram avaliadas as características altura de plantas, altura de espigas, rendimento de grãos e capacidade de expansão. Foram observados menores valores de depressão por endogamia e predominância de efeitos gênicos aditivos para capacidade de expansão em relacão ao rendimento de grãos. As populacões BRS ANGELA e SC 002 apresentaram maior probabilidade de sucesso na obtencão de linhagens com boa capacidade de expansão.

Milk line as an indicator of the harvesting time of three hybrid seeds of corn (Zea mays L. )

O experimento foi conduzido durante o ano agrícola de 1996/97 em área experimental da Cooperativa de Desenvolvimento Econômico e Tecnológico (Coodetec), em Cascavel, Paraná, com o objetivo de avaliar o efeito da época de colheita de três híbridos de milho (Zea mays L.) na qualidade fisiológica das sementes e o uso da linha de solidificação do endosperma como indicativo da maturidade fisiológica das sementes. O plantio foi realizado em 30 de setembro de 1996. Amostras de sementes dos híbridos simples CD 1723 e CD 5501, bem como do híbrido duplo OC 705, foram colhidas em intervalos de quatro dias, durante 59 dias, iniciando-se a colheita no vigésimo terceiro dia após o florescimento feminino. As características avaliadas nas sementes foram o acúmulo de matéria seca, o conteúdo de umidade, a germinação, o vigor (pelos testes de frio e de envelhecimento acelerado), a formação da camada preta e o desenvolvimento da linha de solidificação do endosperma. A colheita dos três híbridos realizada a partir dos 47 dias após o florescimento feminino identificada pela análise conjunta das sete características avaliadas, permitiu obter sementes com elevada qualidade fisiológica. O estádio 4 da linha de solidificação do endosperma demonstrou ser muito útil como indicador da época de colheita mais favorável a obtenção de sementes com elevada qualidade fisiológica, uma vez que diferentes estádios de desenvolvimento da linha de solidificação do endosperma em sementes de milho podem ser facilmente identificados no campo, na ausência de qualquer equipamento especial.

Rendimento de grãos e qualidade de sementes de soja após a aplicação de herbicidas dessecantes / Grain yield and seed quality of soybean after desiccants application

Aplicaçöes dos herbicidas diquat (0,3kg ha-1), paraquat (0,4kg ha-1), amônio-glufosinato (0,5kg ha-1) e carfentrazone-ethyl (30g ha-1) foram efetuadas em pré-colheita (estádio R7,5) na cultura da soja, com a finalidade de observar os efeitos na produtividade e na qualidade das sementes. Näo houve diferença significativa entre os tratamentos nos testes de germinaçäo, de classificaçäo do vigor de plântula e de sanidade, bem como na massa de cem sementes, no grau de umidade e no rendimento de sementes. No entanto, o glufosinato de amônio e a testemunha sem herbicida apresentaram maiores percentagens de plântulas normais no teste de envelhecimento acelerado, em relaçäo aos demais tratamentos

Peroxidase and lipid peroxidation of soybean roots in response to p-coumaric and p-hydroxybenzoic acids

The scope of the present study was to investigate how the p-coumaric (p-CA) and p-hydroxybenzoic (p-HD) acids affect the peroxidase (POD, EC 1.11.1.7) activity, the lipid peroxidation (LP) and the root growth of soybean (Glycine max (L.) Merr.). Three-day-old seedlings were cultivated in nutrient solution containing p-CA or p-HD (0.1 to 1 mM) for 48 h. After uptake, both compounds (at 0.5 and 1 mM) decreased root length (RL), fresh weight (FW) and dry weight (DW) while increased soluble POD activity, cell wall (CW)-bound POD activity (with 1 mM p-CA and 0.5 mM p-HD) and LP

[Cor triatriatum dextrum resulting from the persistence of embryonic remnants of the right valve of the sinus venosus: prevalence and echocardiographic aspects in a large consecutive non-selected patient population]. / Pseudocuore triatriato destro da persistenza di residui embrionari della primitiva valvola del seno venoso: prevalenza ed aspetti ecocardiografici in un'ampia popolazione consecutiva non selezionata.

BACKGROUND: Cor triatriatum destrum is a rare congenital malformation, usually associated with complex right heart abnormalities, characterized by a membrane that divides the right atrium into two chambers. It is considered the result of the incomplete and abnormal regression of the embryonic right valve of the sinus venosus. When the valve regresses the cephalic portion forms the crista terminalis and the caudal portion develops into the Eustachian and Thebesian valve. With an incomplete regression, a fenestrated or an unfenestrated membrane may persist in the right atrium. We describe the clinical and echocardiographic characteristics of this type of remnants in the right atrium. METHODS: In a 4-month period, 1728 transthoracic two-dimensional and Doppler echocardiographic studies were consecutively performed. RESULTS: Twenty-seven patients (16 males, 11 females, mean age 55 +/- 21 years, range 1-87 years) were occasionally recognized to be affected by a membrane inside the right atrial cavity. On the basis of its echocardiographic appearance (a membranous structure connecting the orifice of the inferior vena cava to the atrial septum with evident fenestration in 74% of cases), absence of associated congenital abnormalities (92.5% of cases), interatrial shunt or abnormal flow pattern of venous systemic return, a diagnosis of a normal benign anatomic variant of sinus venous remnants (similar to the so-called "Chiari's network") could be made in every case. CONCLUSIONS: The incomplete regression of the embryonic right valve of the sinus venosus may leave a fenestrated or an unfenestrated membrane in the right atrium that should be considered a normal benign variant of the so-called "Chiari's network".

Sensitization to ethanol's stimulant effect is associated with region-specific increases in brain D2 receptor binding.

RATIONALE: Stimulation of locomotor activity by low doses of ethanol (EtOH) and the potentiation of this response after repeated administration (sensitization) have been related to EtOH's rewarding and addictive properties and to altered dopaminergic activity in brain. In mice, behavioral sensitization to EtOH occurs only in a subset of treated animals, and this provides an opportunity for distinguishing general drug effects from sensitization-specific brain effects. OBJECTIVES: In view of evidence suggesting a role for dopamine D2 receptors in EtOH preference and abuse liability, the present study addressed the hypothesis that D2 binding would be altered in specific brain regions in mice showing differential sensitization responses to chronic EtOH administration. METHODS: Male albino Swiss mice received 2.4 g/kg EtOH i.p. daily for 21 days and were then separated into sensitized or non-sensitized subgroups on the basis of weekly locomotor activity tests. RESULTS: Autoradiographic analyses of [(3)H]raclopride binding to D2 sites revealed significant increases in the anterior caudate-putamen of mice in the EtOH-sensitized group when compared with either saline controls (+40%, P<0.00009) or to mice in the EtOH non-sensitized group (+32%; P<0.0003). Smaller increases were seen in the ventrolateral caudate-putamen of sensitized animals (+18% vs. control, P<0.02; and 12% vs. non-sensitized mice, P<0.07). No differences were found in other brain regions, including the nucleus accumbens, olfactory bulb and substantia nigra. CONCLUSIONS: The observed increases in D2-receptor binding in circumscribed targets of nigrostriatal projections may reflect either a pre-existing condition in sensitization-prone animals or a selective vulnerability of D2 receptors to chronic EtOH in these animals. In either case, it may be a marker for differential susceptibility to EtOH sensitization.

Extracellular calcium deprivation in astrocytes: regulation of mRNA expression and apoptosis.

Cell viability and gene expression were studied in primary astroglial cells cultured in a nominally calcium-free medium. Ca2+ deprivation reduced progressively the astrocytes' viability, starting from 12 h; the restoration of a normal Ca2+ concentration (1.8 mM) in the medium after 12-h deprivation reversed the degenerative effect within 24 h. Biochemical and morphological examinations indicated that cell death induced by Ca2+ deprivation was mediated by apoptosis. This was associated with the expression of c-fos, c-jun, and c-myc, which, with different time courses, were induced in astrocytes after Ca2+ deprivation. Furthermore, shifting to a Ca2+-free medium modified the expression of Ich-1S transcript and rapidly increased intracellular cyclic AMP, which has been implicated in the transcriptional activation of immediate-early genes. The absence of Ca2+ in the medium reduced the expression of constitutive proteins such as alpha-actin, clusterin, glial fibrillary acidic protein, amyloid precursor protein, and glucose-6-phosphate dehydrogenase. The expression of these mRNAs was reduced >50% after 8 h of Ca2+ deprivation, when the effect on cell viability was negligible. When Ca2+ deprivation was prolonged for 24 h the expression of mRNA dropped completely, and restoration of the Ca2+ ions in the medium for 48 h did not reverse this effect. In contrast with general assumption, the apoptotic machinery in astrocytes is activated similarly not only by increased Ca2+ influx but also with the extracellular Ca2+ deprivation.

Densitometric quantification of neuronal viability by computerized image analysis.

A new method is presented for the quantification of cell viability based on densitometry with computerized image analysis. Neuronal cells were stained with crystal violet and densitometric analysis was performed with an IBAS 2.0 image analyzer (Kontron/ Zeiss), using specially implemented dedicated software which integrates the optical density of the culture in each well with the area covered by the stained cells. To test the reliability of the densitometric method cortical cells were plated at different concentrations (5 x 10(4)-10(6)/ml); the standard curve obtained by analysis of crystal violet staining showed a linear proportion between cell number and optical density signal. The validation and accuracy of the method were assessed and compared with other methods using rat cortical cells cultured in vitro for 10 days and exposed to kainic acid (250 microM) for 24 h. Neuronal viability was reduced by 40-50% and comparison with direct cell counting, MTT assay, and spectrophotometric analysis confirmed that the method is simple, quick, and reliable.

Beta-amyloid fragment potentiates IL-6 and TNF-alpha secretion by LPS in astrocytes but not in microglia.

The effect of a peptide homologous to the biologically active fragment of beta amyloid 25-35 (beta 25-35) was studied on interleukin 6 (IL-6) and tumour necrosis factor (TNF-alpha) secretion induced by lipopolysaccharide (LPS) in primary rat astrocytes and microglia. Twenty-four hour exposure to LPS (50 ng/ml) induced IL-6 and TNF-alpha both in astrocytes and in microglial cells, while the effect of beta 25-35 (50 microM) per se was negligible in both cell types. In microglial cells, the application of beta peptide did not alter the production of either cytokine induced by LPS. However, beta 25-35 strongly amplified the production of both IL-6 and TNF-alpha in astrocytes. These findings confirm the complex interaction between cytokines and amyloidogenesis in Alzheimer's disease and indicate that astrocytes rather than microglia respond to the beta amyloid fragment, suggesting that these cells may be actively involved in cytokine-mediated events in AD.

Influence of cell culture conditions on the protective effect of antioxidants against beta-amyloid toxicity: studies with lazaroids.

The mechanisms of cell death of rat cortical neurons chronically exposed to the beta-amyloid (betaA) biologically active fragment beta-(25-35) involve oxidative stress. We examined the influence of culture conditions on the neuroprotective activity of antioxidants against beta-(25-35) toxicity. Common radical scavengers such as N-acetylcysteine (250 microM) and N-t-butyl-phenylnitrone (500 microM) only protected cortical cells cultured in the presence of fetal calf serum (FCS) from betaA insult. The neuroprotective effect of lazaroids (U74389G and U83836E), 21-aminosteroids with antioxidant activity, was tested in cells grown with or without FCS. U74389G did not interfere with beta-(25-35) toxicity in either condition, while U83836E at a very low concentration (15 nM) protected cortical cells exposed to the beta peptide only when the neurons were cultured in the presence of FCS. These data show that a lazaroid can prevent beta-(25-35) toxicity and that the antioxidants exerted their protective effect in certain conditions.

Prion protein fragment 106-126 differentially induces heme oxygenase-1 mRNA in cultured neurons and astroglial cells.

Heme oxygenase (HO), which catalyzes the degradation of heme, has two isozymes (HO-1 and HO-2). In brain the noninducible HO-2 isoform is predominant, whereas the inducible HO-1 is a marker of oxidative stress. Because brain oxidative stress might be present in prion-related encephalopathies (PREs), as in other neurodegenerative diseases, we investigated whether HO-1 mRNA was induced in neuronal and astroglial cell cultures by a peptide corresponding to residue 106-126 of human prion protein (PrP). This peptide is amyloidogenic, and when added in vitro to cultured cells it reproduces the neuronal death and astroglial proliferation and hypertrophy occurring in PREs. HO-1 mRNA did not accumulate in rat cultured neurons from hippocampus or cortex exposed to PrP 106-126 (50 microM for 5 days). PrP 106-126 induced HO-1 mRNA accumulation in rat astroglial cultures depending on the exposure time and concentration, being maximal (33-fold) after 7 days of exposure at 50 microM. The nonamyloidogenic amidated or amidated-acetylated PrP 106-126 was ineffective, as was a scrambled peptide used as control. N-Acetylcysteine reduced (50%) the accumulation of HO-1 mRNA in astroglial cells after PrP 106-126 (25 microM) given for 5 days. Thus, oxidative stress is apparently a feature of the toxicity of PrP 106-126, and it might also occur in PREs; induction of HO-1 could contribute to the greater resistance of astrocytes compared with neurons to PrP 106-126 toxicity.