Reactive oxygen species in inflammation

Chronic inflammation is the underlying cause of many common disabling conditions such as rheumatoid arthritis (RA), multiple sclerosis, coeliac disease, type I diabetes and coronary artery disease. NOX2 complex derived reactive oxygen species (ROS) are known to regulate joint inflammation in rats and mice, and additionally recent genetic evidence associates phagocyte ROS and the development RA in humans. Ncf1mutated mice have lost the functionality of their NOX2 complex and thus have no phagocyte ROS production. These mice suffer from exacerbated arthritis. The immune suppressive effect of the NOX2 complex derived ROS is mediated by monocytes/macrophages that downregulate the activation of autoreactive T cells. The aim of this thesis was to study how ROS modulate immune responses in different arthritis models and in tumor development. Additionally, genome wide gene expression profiling was carried out to assess the global effects of NOX2 complex derived ROS. Firstly, these results confirmed the potent anti-inflammatory nature of phagocyte ROS in arthritis models that were driven by the adaptive immune system. Secondly, arthritis models with predominantly innate immunity induced pathophysiology were moderately enhanced by phagocyte, more specifically, neutrophil derived ROS. Thirdly, the ROS induced immune suppression mediated by the adaptive immune system allowed development of bigger implanted tumors, while phagocyte ROS production did not affect the development of spontaneously growing tumors. Lastly, genome wide gene expression analysis revealed that both humans and mice with abrogated phagocyte NOX2 complex ROS production had an enhanced type I interferon signature in blood, reflecting their hyperinflammatory immune status.

Efeitos de doses e épocas de aplicação do bentazon na produção de duas cultivares de cebola (Allium cepa L.)

Em um solo aluvião eutrófico de classificação textural argilo siltoso, com 2,75% de M.O. foram estudados os efeitos de doses do herbicida bentazon (3 - isopropi l - 2,1,3 - benzotio-diazinona - (4) - 2,2 - dióxido) aplicado em pós-emergência. no controle de plantas daninhas dicotiledôneas na cultura da cebola. O delineamento experimental adotado foi um fatorial 2 x 5 x 2 + 4. constituído por duas cultivares: 'Baia Pe ri forme' e 'Texas Grano': cinco épocas de aplicação do produto: aos 10, 20, 30, 40 e 50 dias após a emergência das plantas daninhas: duas doses de bentazon: 0,46kg. i.a./ha e 0,96 kg. i.a./ha e quatro tratamentos adicionais: 'Baia Periforme' capinada e sem capina e 'Texas Grano'. capinada e sem capina. . As plantas daninhas predominantes no experimento foram: botão -de-ouro (Galinsoga parviflora Cav.), picão-preto (Bidens pilosa L.), amendoim-bravo (Euphorbia prunifoiia Jacq.(, mostarda (Brassica campestris L.), joá (Physalis angulata L.) fedegoso (Cassia tora L.(, serralha falsa (Emilia sonchifolia (L .) DC) e mentruz (Lepidium virginicum L. Nas condições em que foi realizado o ensaio, o herbicida bentazon nas duas doses aplicadas mostrou-se eficiente no controle das plantas daninhas até 60 dias após o transplante. Nas doses empregadas o bentazon não apresentou efeito fitotóxico sobre a produção das cultivares 'Baia Periforme' e 'Texas Grano'.

Efeitos de algumas características do solo na resposta da planta a doses de herbicidas

Foram estudados em casa de vegetação o tempo de sobre vivência de oito espécies olerícolas a aplicação de atrazina nas doses correspondentes a 1,8 e 3,2 kg/ha, e de diuron nas doses de 1,2 e 2,0 kg/ ha, em três tipos de solos ácidos tropicais. Os resultados mostraram alta correlação linear entre o tempo médio de vida das plântulas com teores de matéria orgânica, capacidade de troca catiônica (CTC) efetiva e pH do solo; e baixas correlações com teores de argila mais silte. Os resultados indicam ser mais adequado tomar como base a característica CTC do solo em lugar da sua classificação textural para fins de recomendação das doses dos herbicidas.

Efeitos da calagem do solo no crescimento inicial e absorção de macronutrientes por plantas de trapoeraba (Commelina benghalensis)

A presente pesquisa foi conduzida visando estudar os efeitos da calagem do solo sobre o crescimento inicial e absorção de macronutrientes por plantas de trapoeraba (Commelina benghalensis L.). A fase experimental foi conduzida em casa de vegetação, em vasos de cinco litros e o substrato foi coletado na camada arável de um Latossolo Vermelho Escuro, distrófico, classe textural franco-argilo-arenosa e valor original de pH igual a 3,8. O experimento foi instalado no delineamento inteiramente casualizado com quatro repetições e os tratamentos constaram da incorporação de quantidades correspondentes às doses 0, I, 2, 3, 4 e 5 t/ha de calcário dolomítico calcinado. Foi efetuada uma incubação por 15 dias antes do plantio das mudas da trapoeraba. Os resultados foram avaliados 49 dias após o transplante. A planta daninha respondeu intensamente à calagem incrementando a área foliar e os acúmulos de matéria seca e de N, P, Ca, Mg e S. Os padrões de distribuição dos diferentes macronutrientes nas diversas estruturas morfológicas da planta foram alterados. O acúmulo de K cresceu até a dose de 2 t/ha de calcário e depois decresceu, possivelmente devido ao antagonismo com Mg, adicionado através do calcário.

Seletividade de imazapic às soqueiras de cana-de-açúcar (Saccharum spp.)

Esta pesquisa objetivou a avaliar a seletividade de imazapic às soqueiras de cana-de-açúcar, cultivar RB 835089, comparado a imazapyr e tebuthiuron. O estudo foi conduzido no município de Araras-SP, durante o período de 1999/2000, em solo de classe textural franco-argilo-arenosa, devidamente corrigido e adubado. A instalação do experimento foi em blocos, com sete tratamentos casualizados e cinco repetições, a saber: imazapic (73,5; 98,0; e 122,5 g ha-1), imazapyr (125,0 g ha-1) e tebuthiuron (1.100,0 g ha-1). Os herbicidas foram aplicados em pós-emergência inicial da cultura, além das testemunhas com e sem capinas. Foi possível concluir que todos os produtos testados causaram leves sintomas de intoxicação inicial nas plantas de cana-de-açúcar, com total recuperação aos 100 dias após os tratamentos, sem prejuízo da produtividade e qualidade da matéria-prima.

Seletividade de herbicidas pós-emergentes aplicados na soja geneticamente modificada

O experimento foi instalado em área de plantio comercial de soja Roundup Ready®, na região do Pontal do Paranapanema, no município de Euclides da Cunha Paulista-SP, localizada a 20º 43' 11'' S e 50º 10' 20'' W, com uma altitude de 270 m. A fase experimental foi conduzida de dezembro de 2006 a abril de 2007, sob sistema plantio direto, com uma temperatura média de 25 ºC e índice pluviométrico de 800 mm. O solo da área experimental apresenta classe textural franco-argilo arenosa. Objetivou-se com este trabalho avaliar a eficácia e seletividade de glyphosate na formulação Roundup Transorb®, associado aos herbicidas diclosulam, cloransulam-methyl, flumioxazina e S-metolachlor em duas modalidades de aplicação: única, com associação do glyphosate aos herbicidas; e uma sequencial apenas com glyphosate aos 15 DAA, no manejo das plantas daninhas trapoeraba (Commelina benghalensis) e corda-de-viola (Ipomoea triloba) durante o cultivo da soja. O experimento foi instalado no delineamento de blocos ao acaso, com 12 tratamentos e quatro repetições. Os tratamentos foram distribuídos em arranjo fatorial acrescido de duas testemunhas: sem capina manual e com capina manual. O arranjo fatorial 2 x 5 contemplou duas condições de aplicação dos herbicidas (única e sequencial) e cinco herbicidas (glyphosate, glyphosate + diclosulam, glyphosate + cloransulam-methyl, glyphosate + flumioxazin e glyphosate + S-metolachlor). Nas condições de produtos, épocas de aplicação e doses, os resultados mostraram que o herbicida glyphosate aplicado em dose única ou sequencial e suas combinações com diclosulam e cloransulam-methyl na primeira aplicação não promovem fitointoxicação nas plantas de soja. A combinação com flumioxazin e S-metolachlor promoveu atraso no crescimento das plantas e no fechamento da cultura, em razão do efeito na altura dos indivíduos e comprimento dos ramos. No tratamento com o S-metolachlor, isso pode ser explicado pelo fato de o herbicida ser registrado para controle em pré-emergência e ter sido aplicado em pós-emergência. Nenhum dos tratamentos influenciou significativamente a produção de grãos da cultura da soja. A aplicação única ou a complementação com aplicação sequencial de glyphosate promoveram excelente controle de Commelina benghalensis e Ipomoea triloba.

Controle de plantas daninhas pelo indaziflam em solos com diferentes características físico-químicas

No solo, os herbicidas estão sujeitos a processos de degradação e sorção, que regulam seu destino e sua disponibilidade. Objetivou-se com este trabalho avaliar a eficiência do indaziflam no controle de cinco espécies de plantas daninhas em três solos com características físico-químicas contrastantes. O experimento foi conduzido em casa de vegetação, no delineamento de blocos ao acaso, arranjado em esquema fatorial 6 x 3 x 5, com quatro repetições. Os tratamentos constaram de seis doses de indaziflam (0, 30, 60, 90, 120 e 150 g i.a. ha-1), três solos (texturas areia, argila e franco-argiloarenosa) e cinco espécies daninhas (Rottboellia cochinchinensis, Panicum maximum, Digitaria horizontalis, Euphorbia heterophylla e Ipomoea grandifolia). O solo foi peneirado e colocado em bandejas perfuradas, e as espécies, semeadas a 0,015 m de profundidade. O herbicida foi aplicado sobre solo úmido, e as bandejas, colocadas sob irrigação diária de 5 mm, aproximadamente. Avaliou-se a emergência, sete dias após a emergência, e o controle das espécies, 40 dias depois da aplicação. O indaziflam foi mais eficiente no controle das espécies D. horizontalis, P. maximum e R. cochinchinensis. As duas primeiras foram muito sensíveis ao herbicida, com controle total dessas espécies em todas as doses e solos testados. No solo argiloso, R. cochincinensis só foi eficientemente controlada a partir da dose de 50 g ha-1 de indaziflam. O controle de E. heterophylla e I. grandifolia pelo indaziflam foi mais eficiente no solo franco-argiloarenoso.

SELECTION OF HERBICIDES TARGETING THE USE IN CROP SYSTEMS CULTIVATED WITH SHOWY CROTALARIA

ABSTRACT The increase in the area planted with Crotalaria spectabilishas occurred by several factors, highlighting the potential to reduce the nematodes, nitrogen fixation and the high production of biomass. By becoming a species sown as a crop, it is necessary to control the weeds that coexist with showy crotalaria. This change in the use of this crop creates the possibility of this specie becoming a weed. The aim of this study was to assess the potential use of herbicides applied in preemergence and postemergence of C.spectabilisfor different purposes (control of volunteer and selectivity plants). Three experiments were installed in a greenhouse (two with herbicides applied in preemergence - in soils with distinct textural categories; and one experiment with herbicides applied in postemergence). The results of the experiments with herbicides applied in preemergence showed that: amicarbazone, atrazine, diuron, metribuzin, prometryn, fomesafen and sulfentrazone showed effectiveness for control of C.spectabilis in clayey soil. Besides these, flumioxazin and isoxaflutole also showed potential to be used in the control of showy crotalaria in soils with loam texture. In relation to the postemergence herbicides, atrazine, diuron, prometryn, flumioxazin, fomesafen, lactofen, saflufenacil, amonio-glufosinate and glyphosate can be used aiming the chemical control of C.spectabilis. Herbicides chlorimuron-ethyl, diclosulan, imazethapyr, pyrithiobac-sodium, trifloxysulfuron-sodium, clomazone, pendimethalin, S-metolachlor and trifluralin applied in preemergence, and imazethapyr, pyrithiobac-sodium, flumiclorac, bentazon and clethodim applied in postemergence caused low levels of injury to C.spectabilis plants, making necessary the development of new searches to ensure the selectivity of these products.

Image analysis and definition of nuclear phenotypes

Some basic topics concerned with the extraction of textural and geometric information from cell nucleus images as well as description and characterization of chromatin supraorganization and consequent classification of nuclear phenotypes are presented.

P-glycoprotein-mediated resistance to chemotherapy in cancer cells: using recombinant cytosolic domains to establish structure-function relationships

Resistance to chemotherapy in cancer cells is mainly mediated by overexpression of P-glycoprotein (Pgp), a plasma membrane ATP-binding cassette (ABC) transporter which extrudes cytotoxic drugs at the expense of ATP hydrolysis. Pgp consists of two homologous halves each containing a transmembrane domain and a cytosolic nucleotide-binding domain (NBD) which contains two consensus Walker motifs, A and B, involved in ATP binding and hydrolysis. The protein also contains an S signature characteristic of ABC transporters. The molecular mechanism of Pgp-mediated drug transport is not known. Since the transporter has an extraordinarily broad substrate specificity, its cellular function has been described as a "hydrophobic vacuum cleaner". The limited knowledge about the mechanism of Pgp, partly due to the lack of a high-resolution structure, is well reflected in the failure to efficiently inhibit its activity in cancer cells and thus to reverse multidrug resistance (MDR). In contrast to the difficulties encountered when studying the full-length Pgp, the recombinant NBDs can be obtained in large amounts as soluble proteins. The biochemical and biophysical characterization of recombinant NBDs is shown here to provide a suitable alternative route to establish structure-function relationships. NBDs were shown to bind ATP and analogues as well as potent modulators of MDR, such as hydrophobic steroids, at a region close to the ATP site. Interestingly, flavonoids also bind to NBDs with high affinity. Their binding site partly overlaps both the ATP-binding site and the steroid-interacting region. Therefore flavonoids constitute a new promising class of bifunctional modulators of Pgp.

Thermodynamic evaluation and modeling of proton and water exchange associated with benzamidine and berenil binding to ß-trypsin

Serine-proteases are involved in vital processes in virtually all species. They are important targets for researchers studying the relationships between protein structure and activity, for the rational design of new pharmaceuticals. Trypsin was used as a model to assess a possible differential contribution of hydration water to the binding of two synthetic inhibitors. Thermodynamic parameters for the association of bovine ß-trypsin (homogeneous material, observed 23,294.4 ± 0.2 Da, theoretical 23,292.5 Da) with the inhibitors benzamidine and berenil at pH 8.0, 25ºC and with 25 mM CaCl2, were determined using isothermal titration calorimetry and the osmotic stress method. The association constant for berenil was about 12 times higher compared to the one for benzamidine (binding constants are K = 596,599 ± 25,057 and 49,513 ± 2,732 M-1, respectively; the number of binding sites is the same for both ligands, N = 0.99 ± 0.05). Apparently the driving force responsible for this large difference of affinity is not due to hydrophobic interactions because the variation in heat capacity (DCp), a characteristic signature of these interactions, was similar in both systems tested (-464.7 ± 23.9 and -477.1 ± 86.8 J K-1 mol-1 for berenil and benzamidine, respectively). The results also indicated that the enzyme has a net gain of about 21 water molecules regardless of the inhibitor tested. It was shown that the difference in affinity could be due to a larger number of interactions between berenil and the enzyme based on computational modeling. The data support the view that pharmaceuticals derived from benzamidine that enable hydrogen bond formation outside the catalytic binding pocket of ß-trypsin may result in more effective inhibitors.

Evaluation of thermotolerant capacity of lactic acid bacteria isolated from commercial sausages and the effects of their addition on the quality of cooked sausages

The thermotolerant capacity of several lactic acid bacteria strains isolated from cooked commercial sausages was determined. Four strains were positively identified as Lactobacillus plantarum, Lactobacillus curvatus, Pediococcus pentosaceus and Pediococcus acidilacti, after surviving thermal treatment (70 °C during 60 minutes). Thermotolerant strains were inoculated in sausage batters before cooking in order to determine their effect on color, texture, acceptance and inhibition of Enterobacteria during 12 days at 8 °C. No significant effect of the inoculated strains was detected on color parameters. Textural profile parameters, cohesiveness and resilience, were not affected by the inoculation of thermotolerant lactic acid bacteria, but L. curvatus sausages resulted softer than the rest of the treatments. Samples inoculated with L. curvatus also obtained the lowest scores for the sensory attributes evaluated, with the remaining treatments causing no unfavorable effects on sausage acceptance. There was a reduction in enterobacterial counts after 12 days of cold storage in inoculated samples. The performance of inoculated lactic acid bacteria strains can be explained in a similar way as that of starter cultures in dry-fermented sausages, where the growth in nests impairs other pathogenic microorganisms present in the rest of the sausage, since environmental conditions and the early inoculation of these thermotolerant strains favor them to become the dominant flora.

Quality of minimally processed guava with different types of cut, sanification and packing

The purpose of this project was to evaluate the sanitization effect on the quality of minimally processed guava. Initially, research was carried out with consumers in a supermarket to verify preferences of packaging for guava. Following this, the guava cv. Paluma underwent two sanitization sequences using dehydrated sodium dichloroisocyanurate compound, in 50 ppm concentration, sanitization prior to (S1) and after (S2) being cut; removal of excess water; conditioning in PET packaging and PSPVC and storage at 3 ºC ± 1 ºC. Physicochemical analysis - [pH, total soluble solids (SST), total labeled acidity (ATT), ascorbic acid (AA), total sugars (AT) and reducers (AR)], textural sensorial and microbiological analyses were used to monitor the quality of the products. The consumers preferred the guava cut in halves with pulp and packed in PET, although this packaging promoted condensation of water vapor on the inner surface of the lid, compromising the appearance of the product. The two sanitization sequences and the two kinds of packaging did not significantly affect the pH, SST, ATT, SST/ATT, texture and AA values. The AT and AR tenors increased significantly in the MP guavas stored in the PSPVC package. Both sanitizations were efficient in the bacterial control of the indicators of the hygienicsanitary conditions, although the S1 sanitization proved to be more efficient in the control of autochthonous aerobic microbiota (aerobic mesophylic microorganisms). It can be concluded that guava cv. Paluma packed in PSPVC can be conserved for 6 days when stored at 3 ºC.

Effect of wheat flour protein variations on sensory attributes, texture and staling of Taftoon bread

The quality of flat breads depends in part on the textural properties of breads during storage. These properties are largely affected by flour protein quality and quantity. The present study aimed to examine differences between sensory properties, textural and staling of Tandoori breads made from flours of different quality and different quantities of protein. This was implemented by using three flours with 9.4, 11.5 and 13.5% protein contents and different protein qualities shown by Zeleney sedimentation volume 16.25, 22.75 and 23.25 mL respectively. Bread strips were submitted to uniaxial compression between two parallel plates on an Instron Universal Testing machine, and firmness of the breads was determined. Results indicated the differences in the sensory attributes of breads produced by flours of different protein content and quality, demonstrating that high protein high quality flours are not able to sheet and expand under the high temperature - short time conditions employed in Taftoon bread production and are therefore not suitable for this kind of bread. Results showed that flour with 11.5% protein content, produced bread with better sensory characteristics and acceptable storage time.

Partial substitution of nitrite by chitosan and the effect on the quality properties of pork sausages

The aim of the study was to evaluate the influence of partial nitrite replacement by chitosan on the quality of Ham Visking (a type of pork sausages). Five Ham Visking formulations were elaborated modifying the sodium nitrite (0.011; 0.016 or 0.0212%) and chitosan concentrations (0.25 or 0.5%) in the products. Sausages were stored at 4 ºC and physicochemical, microbiological, and sensorial evaluations were performed in order to estimate their shelf life. Chitosan can be used in pork sausages without affecting ensory attributes such as color although the panelists detected textural differences among the samples with chitosan, which suggests that there is some influence of deacetylation degree of chitosan on the textural behavior of sausages which still needed to be explained for a successful application of chitosan in meat products. The reduction of residual sodium nitrite did not affect the color and flavor of such products, but the use of chitosan increasedsignificantly the shelf life of sausages.