Efeitos da microgravidade em plantas de cana-de-açúcar

Sugarcane (Saccharum spp.) is a plant from Poaceae family that has an impressive ability to accumulate sucrose in the stalk, making it a significant component of the economy of many countries. About 100 countries produce sugarcane in an area of 22 million hectares worldwide. For this reason, many studies have been done using sugarcane as a plant model in order to improve production. A change in gravity may be one kind of abiotic stress, since it generates rapid responses after stimulation. In this work we decided to investigate the possible morphophysiological, biochemical and molecular changes resulting from microgravity. Here, we present the contributions of an experiment where sugarcane plants were submitted to microgravity flight using a vehicle VSB-30, a sounding rocket developed by Aeronautics and Space Institute teams, in cooperation with the German Space Agency. Sugarcane plants with 10 days older were submitted to a period of six minutes of microgravity using the VSB-30 rocket. The morphophysiological analyses of roots and leaves showed that plants submitted to the flight showed changes in the conduction tissues, irregular pattern of arrangement of vascular bundles and thickening of the cell walls, among other anatomical changes that indicate that the morphology of the plants was substantially influenced by gravitational stimulation, besides the accumulation of hydrogen peroxide, an important signaling molecule in stress conditions. We carried out RNA extraction and sequencing using Illumina platform. Plants subjected to microgravity also showed changes in enzyme activity. It was observed an increased in superoxide dismutase activity in leaves and a decreased in its activity in roots as well as for ascorbate peroxidase activity. Thus, it was concluded that the changes in gravity were perceived by plants, and that microgravity environment triggered changes associated with a reactive oxygen specie signaling process. This work has helped the understanding of how the gravity affects the structural organization of the plants, by comparing the anatomy of plants subjected to microgravity and plants grown in 1g gravity

Proteômica diferencial da Chromobacterium violaceum em resposta ao estresse oxidativo induzido por peróxido de hidrogênio

The β-proteobacterium Chromobacterium violaceum is a Gram-negative, free-living, saprophytic and opportunistic pathogen that inhabits tropical and subtropical ecosystems among them, in soil and water of the Amazon. It has great biotechnological potential, and because of this potential, its genome was completely sequenced in 2003. Genome analysis showed that this bacterium has several genes with functions related to the ability to survive under different kinds of environmental stresses. In order to understand the physiological response of C. violaceum under oxidative stress, we applied the tool of shotgun proteomics. Thus, colonies of C. violaceum ATCC 12472 were grown in the presence and absence of 8 mM H2O2 for two hours, total proteins were extracted from bacteria, subjected to SDS-PAGE, stained and hydrolysed. The tryptic peptides generated were subjected to a linear-liquid chromatography (LC) followed by mass spectrometer (LTQ-XL-Orbitrap) to obtain quantitative and qualitative data. A shotgun proteomics allows to compare directly in complex samples, differential expression of proteins and found that in C. Violaceum, 131 proteins are expressed exclusively in the control condition, 177 proteins began to be expressed under oxidative stress and 1175 proteins have expression in both conditions. The results showed that, under the condition of oxidative stress, this bacterium changes its metabolism by increasing the expression of proteins capable of combating oxidative stress and decreasing the expression of proteins related processes bacterial growth and catabolism (transcription, translation, carbon metabolism and fatty acids). A tool with of proteomics as an approach of integrative biology provided an overview of the metabolic pathways involved in the response of C. violaceum to oxidative stress, as well as significantly amplified understanding physiological response to environmental stress. Biochemical and "in silico" assays with the hypothetical ORF CV_0868 found that this is part of an operon. Phylogenetic analysis of superoxide dismutase, protein belonging to the operon also showed that the gene is duplicated in genome of C. violaceum and the second copy was acquired through a horizontal transfer event. Possibly, not only the SOD gene but also all genes comprising this operon were obtained in the same manner. It was concluded that C. violaceum has complex, efficient and versatile mechanisms in oxidative stress response

Estudo de adsorvente obtido de terras diatomáceas para remoção de metais pesados em águas produzidas: processamento, caracterização e dinâmica em leito fixo

The acceleration of industrial growth in recent decades on all continents aroused the interest of the companies to counter the impacts produced on the environment, spurred primarily by major disasters in the petroleum industry. In this context, the water produced is responsible for the largest volume of effluent from the production and extraction of oil and natural gas. This effluent has in its composition some critical components such as inorganic salts, heavy metals (Fe, Cu, Zn, Pb, Cd, ), presence of oil and chemicals added in the various production processes. In response to impact, have been triggered by research alternative adsorbent materials for water treatment and water produced, in order to removing oils and acids and heavy metals. Many surveys of diatomaceous earth (diatomite) in Brazil involve studies on the physico-chemical, mineral deposits, extraction, processing and applications. The official estimated Jazi are around 2.5 million tonnes, the main located in the states of Bahia (44%) and Rio Grande do Norte (37,4%). Moreover, these two states appear as large offshore producers, earning a prominent role in research of adsorbents such as diatomite for treatment of water produced. Its main applications are as an agent of filtration, adsorption of oils and greases, industrial load and thermal insulator. The objective of this work was the processing and characterization of diatomite diatomaceous earth obtained from the municipality of Macaíba-RN (known locally as tabatinga) as a low cost regenerative adsorbent for removal of heavy metals in the application of water produced treatment. In this work we adopted a methodology for batch processing, practiced by small businesses located in producing regions of Brazil. The characterization was made by X-ray diffraction (XRD), scanning electron microscopy (SEM) and specific surface area (BET). Research conducted showed that the improvement process used was effective for small volume production of diatomite concentrated. The diatomite obtained was treated by calcination at temperature of 900 oC for 2 hours, with and without fluxing Na2CO3 (4%), according to optimal results in the literature. Column adsorption experiments were conducted to percolation of the in nature, calcined and calcined fluxing diatomites. Effluent was used as a saline solution containing ions of Cu, Zn, Na, Ca and Mg simulating the composition of produced waters in the state of Rio Grande do Norte, Brazil. The breakthrough curves for simultaneous removal of copper ions and zinc as a result, 84.3% for calcined diatomite and diatomite with 97.3 % for fluxing. The calcined fluxing diatomite was more efficient permeability through the bed and removal of copper and zinc ions. The fresh diatomite had trouble with the permeability through the bed under the conditions tested, compared with the other obtained diatomite. The results are presented as promising for application in the petroleum industry

Avaliação dos parâmetros de estresse oxidativo no lúpus eritematoso

OBJECTIVE: The aim of this work was to analyse some oxidative stress parameters in patients of Systemic Lúpus Erythematosus. PATIENTS AND METHODS: Determinations of reduced glutathione content in whole blood were carried out. The activity of superoxide dismutase, gluthatione peroxidase and catalase in erythrocytes and the concentration of reactive substances of acid thiobarbituric in plasma of patients female (n =19) with SLE no activity of disease (Mex-SLEDAI < 2), with average ages of 32 ± 11 years, through the spectrophotometrical methods and from healthy individuals (n =30). Statistical data were analyzed by student t-test, p<0,05. RESULTS: Our data indicated a significant decrease on the activity of catalase and significant increase on the concentration of reactive substances of acid thiobarbituric in patients with SLE comparing with healthy individuals. There was no significant difference in other parameters. CONCLUSION: The results showed that oxidative stress has a role in the pathogenesis of the disease in SLE, even in patients without active disease.

Biochemical and histological characterization of tomato mutants

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

Influência do hipoestrogenismo por ooforectomia na adaptação antioxidante em ratas wistar exercitadas regularmente

Post-menopause is a period of women s life cycle that is characterized by estrogen depletion and therefore increasing cardiovascular diseases, neurodegenerative disorders, urogenital atrophy, osteoporosis, hot flushes and sexual discomfort incidences. Estrogen is a hormone with comfirmed antioxidant action and its depletion is related to oxidative stress instalation and damaging various important biomolecules. Regular physical activity has been identified as a factor involved in reducing women s post-menopausal complications in addition to improving antioxidant defense by reducing the oxidative damage and consequently improving life s quality in this part of the population. This study aims to evaluate the influence of hypoestrogenism in antioxidant adaptation due to regular exercise, by determining reduced glutathione (GSH) and Thiobarbituric Acid Reactive Substances (SRAT) concentrations and antioxidant enzymes glutathione peroxidase (GPx), Superoxide Dismutase (SOD) and Catalase (CAT) activities in blood, brain and liver of rats. To achieve this goal we used 50 Wistar rats, weighing 180-250g which were divided into two groups, control - GC (25) and ooforectomized - GO (25). Each group was subdivided into five subgroups: Not-trained - S (5), Not-trained Acute Exercise - SEA (5), regular exercise 30 days - E30 (5), regular exercise 60 days - E60 (5) and regular exercise 90 days - E90 (5). Each of the three subgroups exercised regularly was subjected to acute exercise on the eve and the day of sacrifice to collect biological samples of blood, liver and brain and subsequent determination of SRAT concentration, GSH content and antioxidant enzymes GPx, SOD and CAT activities. The results indicated that the sedentary animals acutely exercised presented oxidative stress and regular physical activity led to antioxidant adaptation. In ooforectomized group the antioxidant adaptation seen in control animals showed to be impaired. Unlike the results from blood and liver, in brain there was a shield against oxidative damage originated by the exercise and that hypoestrogenism led to a loss of this natural antioxidant potential. Therefore, hypoestrogenism interferes negatively in antioxidant adaptation due to regular exercise

Avaliação do status antioxidante, expressão gênica e polimorfismos dos genes SOD1, SOD2 e GPx1 em crianças, adolescentes e adultos jovens com diabetes tipo 1

Studies report that the pathophysiological mechanism of diabetes complications is associated with increased production of Reactive Oxygen Species (ROS)-induced by hyperglycemia and changes in the capacity the antioxidant defense system. In this sense, the aim of this study was to evaluate changes in the capacity of antioxidant defense system, by evaluating antioxidant status, gene expression and polymorphisms in the genes of GPx1, SOD1 and SOD2 in children, adolescents and young adults with type 1 diabetes. We studied 101 individuals with type 1 diabetes (T1D) and 106 normoglycemic individuals (NG) aged between 6 and 20 years. Individuals with type 1 diabetes were evaluated as a whole group and subdivided according to glycemic control in DM1G good glycemic control and DM1P poor glycemic control. Glycemic and metabolic control was evaluate by serum glucose, glycated hemoglobin, triglycerides, total cholesterol and fractions (HDL and LDL). Renal function was assessed by measurement of serum urea and creatinine and albumin-to-creatinine ratio (ACR) in spot urine. Antioxidant status was evaluate by content of reduced glutathione (GSH) in whole blood and the activity of erythrocyte enzymes glutathione peroxidase (GPx) and superoxide dismutase (SOD). We also analyzed gene expression and gene polymorphisms of GPx1 (rs1050450), SOD1 (rs17881135) and SOD2 (rs4880) by the technique of real-time PCR (Taqman®). Most individuals with DM1 (70.3%) had poor glycemic control (glycated hemoglobin> 8%). Regarding the lipid profile, individuals with type 1 diabetes had significantly elevated total cholesterol (p <0.001) and LDL (p <0.000) compared to NG; for triglycerides only DM1NC group showed significant increase compared to NG. There was an increase in serum urea and RAC of individuals with DM1 compared to NG. Nine individuals with type 1 diabetes showed microalbuminuria (ACR> 30 mg / mg). There was a decrease in GSH content (p = 0.006) and increased erythrocyte GPx activity (p <0.001) and SOD (p <0.001) in DM1 group compared to NG. There was no significant difference in the expression of GPx1 (p = 0.305), SOD1 (.365) and SOD2 (0.385) between NG and DM1. The allele and genotype frequencies of the polymorphisms studied showed no statistically significant difference between the groups DM1 and NG. However, the GPx1 polymorphism showed the influence of erythrocyte enzyme activity. There was a decrease in GPx activity in individuals with type 1 diabetes who had a polymorphic variant T (p = 0.012). DM1 patients with the polymorphic variant G (AG + GG) for polymorphism of SOD2 (rs4880) showed an increase in the RAC (p <0.05). The combined data suggest that glucose control seems to be the predominant factor for the emergence of changes in lipid profile, renal function and antioxidant system, but the presence of the polymorphisms studied may partly contribute to the onset of complications

Nitrogênio, fósforo, potássio, adubo orgânico e calcário dolomítico na produção de sementes de grama-batatais (Paspalum notatum Flügge) em Latossol Vermelho Escuro

Comparou-se a produção de sementes de grama-batatais em Latossol vermelho Escuro com ou sem adição de sulfato de amônio (4g de N/m²), superfosfato simples (50g de P2O5/m²), cloreto de potássio (15g de K2O/m²), esterco de equino (10 litros/m²) e calcário dolomítico (220 g/m²). As produções foram avaliadas através de número de espigas por área, peso total de sementes, peso de sementes puras, relação entre peso de sementes puras e peso total de sementes, peso de 1000 sementes puras, peso total de sementes por espiga, peso de sementes puras por espiga, porcentagem de germinação das sementes e velocidade de emergência das plântulas. Foram determinados os teores de nitrogênio, fósforo, potássio, cálcio, magnésio, enxofre, boro, cobre, ferro, manganês e zinco na matéria seca de folhas e sementes. Verificou-se que a produção de sementes, tanto em quantidade como em qualidade, foi influenciada pela nutrição mineral. As porcentagens mais elevadas de carbono no solo estiveram associadas a maior quantidade e melhor qualidade das sementes produzidas. As melhores produções de sementes correspondem aos níveis mais baixos de fósforo, potássio, cálcio e zinco nas folhas. Verificou-se serem importantes nas características avaliadas as seguintes relações entre teores de nutrientes nas folhas ou nas sementes: N/P, P/Ca, P/Mg, P/S, P/Zn, K/Ca, K/Mg, K/B, Ca/Mg, Ca/B, Fe/Mn, Zn/Cu, Zn/Fe e Zn/Mn. Nas maiores produções, as sementes apresentaram concentrações menores de nitrogênio, boro e cobre, e mais elevadas de cálcio. As sementes de melhor qualidade apresentaram maiores concentrações de zinco e menores de boro. A aplicação de adubos ou calcário dolomítico não melhorou significativamente a produção de sementes.

Produção de mudas de caramboleiras 'B-10' e 'Golden Star': II - marcha de absorção e acúmulo de nutrientes

A participação do Brasil no mercado externo de frutas tem aumentado consideravelmente e com potencial para crescer ainda mais. A constante ascensão dos dados de exportação brasileira é resultado da combinação de avanços tecnológicos do setor produtivo e de acesso a novos mercados consumidores. A caramboleira apresenta-se como uma excelente opção de cultivo de frutas exóticas, com grande potencial para atender ao mercado interno e às exportações. Assim, objetivou-se avaliar a marcha de absorção e de acúmulo de nutrientes em mudas de caramboleiras cultivadas em solução nutritiva. O experimento foi realizado em parcelas subdivididas, sendo utilizadas como parcela as duas cultivares de caramboleira ('B-10' e 'Golden Star') e, como subparcelas, cinco épocas de coleta de plantas, realizadas aos 208; 233; 258; 283 e 308 dias após o transplantio para a solução nutritiva. O delineamento foi inteiramente casualizado, com três repetições. As mudas foram cultivadas em vasos (8L) com solução nutritiva (pH=5,5 ± 0,5), com aeração. O experimento iniciou-se em 24-08-2005. Nos diferentes órgãos das mudas (folhas, caule e raízes), determinaram-se a marcha de absorção, o acúmulo de nutrientes e os índices nutricionais. Não houve diferenças no acúmulo de nutrientes entre as mudas de caramboleira de ambas as cultivares, sendo a ordem decrescente dos nutrientes em cada muda de 'B-10', no final do período experimental: N > K > Ca > Mg > S > P > Fe > Mn > B > Cu > Zn. Para a 'Golden Star', a ordem foi: N > K > Ca > Mg > P > S > Fe > Mn > B > Cu > Zn. Para as duas cultivares, o acúmulo médio foi maior nas folhas > caule > raízes. O período de maior exigência para 'B-10' foi entre 208 - 233 e, para 'Golden Star', entre 233 - 283 dias após o transplantio. As diferentes taxas de acumulação líquida dos nutrientes, nos diferentes órgãos da caramboleira, nem sempre acompanharam a taxa de acumulação de nutrientes do respectivo órgão.

Hypolipemic and antioxidant activities from Tamarindus indica L. pulp fruit extract in hypercholesterolemic hamsters

Dietary modifications may significantly reduce cardiovascular disease (CVD) risk factors, including cholesterol and atherosclerosis. The present study addressed the effects of the crude extract from the pulp fruit of Tamarindus indica L. on lipid serum levels and early atherosclerotic lesions in hypercholesterolemic hamsters in vivo, and the extract's antioxidant action, in vitro. Animals were fed on either chow or atherogenic diet during 10 weeks and concomitantly received either water or T indica L. extract for drinking. Treatment of hypercholesterolemic hamsters with the T. indica pulp fruit extract (5%) led to a decrease in the levels of serum total cholesterol (50%), non-HDL cholesterol (73%) and triglyceride (60%), and to an increase of high-density lipoprotein (HDL) cholesterol levels (61%). In vitro, the extract presented radical scavenging ability, as assessed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide radicals assays, and led to decreased lipid peroxidation in serum, as assessed by the thiobarbituric acid reactive substances (TBARS) assay. In vivo, the extract improved the efficiency of the antioxidant defense system, as assessed by the superoxide dismutase, catalase and glutathione peroxidase activities. Together these results indicate the potential of tamarind extracts in diminishing the risk of atherosclerosis development in humans. (c) 2005 Elsevier Ltd. All rights reserved.

Chemical and physical attributes of two substratum for eucalypt seedling production

This work evaluated the viability of one commercial substrate with pinus bark, vermiculite and 30% of coconut fiber for Eucalyptus grandis seedlings production under different water management: 6, 8, 10, 12 e 14 mm day(-1). In relation to the substrates, it was evaluated, before and after the seedlings production, the total content of carbon, nitrogen and organic substances, pH and EC of the extract, the content of macronutrients (P, K, Ca, S, Mg) and macronutrients (B, Fe, Mn, Cu, Zn), as well as total porosity, density and water retention. It was evaluated the macro and mcronutrients accumulation in the roots and in the aerial part of the seedlings, 105 days after sowing. It was concluded that the substrates presented some distinct properties of those cited as ideal for production in containers with irrigation and fertilization. At the end of the production process, both substrates registered nutrient content little influenced by water management what would guarantee the initial development of the plants in the field.

Características físicas e acúmulo de nutrientes pelos cachos de 'niagara rosada' em vinhedos na região de Jundiaí

Este trabalho teve por objetivo realizar um levantamento em 40 vinhedos de 'Niagara Rosada' nos municípios de Jundiaí e Louveira-SP, sendo metade sobre o porta-enxerto 'Ripária do Traviú' e outra sobre o 'IAC 766'. Determinaram-se, em cada vinhedo, a produtividade, a duração do ciclo, o número de cachos e de ramos produtivos por planta, as características físicas dos cachos e bagas, e o acúmulo de nutrientes pelos cachos. Avaliaram-se nos cachos amostrados a massa da matéria fresca, o comprimento e a largura dos cachos e baga, e diâmetro do pedicelo. Os cachos de cada vinhedo amostrado foram secos em estufa e posteriormente submetidos à análise química para determinar os teores de macro e micronutrientes, visando a estimar o acúmulo de nutrientes por tonelada de uva. Constatou-se um comportamento semelhante entre os porta-enxertos para os dados de produtividade, duração do ciclo, número de cachos e ramos produtivos, sendo de, respectivamente, 11.100kg ha-1, 134 dias, 13 cachos e 9 ramos produtivos. Quanto às características físicas, os valores médios da massa da matéria fresca, comprimento e largura dos cachos foram de, respectivamente, 209g, 12 e 6,8cm; massa da matéria fresca, comprimento e largura das bagas, e diâmetro do pedicelo de, respectivamente, 4,3g; 19,9; 18,2 e 3,2mm. Quanto ao acúmulo de nutrientes, concluiu-se que a cultivar Niagara Rosada enxertada sobre 'Ripária do Traviú' apresentou maior acúmulo de P, Fe e Zn, enquanto sobre o porta-enxerto 'IAC 766' houve maior acúmulo de Mn. em ambos os porta-enxertos, a 'Niagara Rosada' apresentou a seguinte escala de acúmulo de nutrientes em ordem decrescente: K>N>P>Ca>S>Mg>B>Fe>Mn>Cu>Zn.

Assessment of micronutrient extractants from soils of Paraná, Brazil

In Brazil, plant-available micronutrients in the soil can be determined by several chemical extractants, the most common of which are dilute acid and chelating solutions. The purpose of this study was to assess the extractants 0.1 mol L-1 HCl, Mehlich-1, Mehlich-3 and DTPA for analysis of the micronutrients Cu, Zn, Fe, and Mn in soils from the state of Parana. In samples from 12 soils (0-20 cm layer), wheat was planted (Triticum aestivum), grown for 42 days after emergence, and then bean (Phaseolus vulgaris) for 38 days. At the end of each planting period, the soil was sampled again. All extractants tested to assess the availability of Cu, Zn, Fe, and Mn correlated with each other. The efficiency of the extractants HCl, Mehlich-3 and DTPA in assessing plant-available Cu was similar, unlike Mehlich-1, which proved less efficient. The extractants HCl, Mehlich-1 and Mehlich-3 were less efficient in estimating plant-available Zn and Fe, and the most indicated extractant is DTPA. The efficiency of the extractants HCl, Mehlich-1, Mehlich-3 and DTPA in assessing plant-available Mn in soils from Parana was similar.

Efeito da adubação fosfata sobre o acúmulo de biomassa e teor de brix de duas variedades de cana-de-açúcar

The objective of this study was to evaluate the accumulation of matter, dry matter content of Brix and two varieties of sugar cane, influenced by the phosphorus cultivated on "Zona da Mata" region, State of Alagoas. The experiment, a factorial 2 x 6, was composed of two varieties of cane; RB867515 and RB92579 and six phosphorus levels: zero, 30, 60, 90, 120 and 150 kg ha(-1), with the source of the phosphorus triple superphosphate. Was also used nitrogen and potassium in doses equivalent to 100 and 200 kg ha(-1) and K, respectively. The soil of the experimental area was set to raise the base saturation to 60%. At the beginning of the first rainfall of the rainy season of 2006 was performed with micro fertilization in coverage, in doses equivalent to 6,0; 6,0 and 7,0 kg ha(-1) of Cu, Zn and Mn, respectively. Was The chemical control of weeds. Every month, from August to October 2006, samples were collected in the cane plant. There was initially the number of plants m(2), followed by weighing the material. It was subsequently withdrawn a subsample containing ten plants for weighing. After weighing the material was passed in horsemanship and homogenized, again drew up a sample of approximately 300g, leading them to forced ventilation oven at 65 degrees C, toobtain constant weight. Resulting in the production of natural (MN) and dry matter (DM). In July, we analyzed the levels of Brix% juice in the refractometer Brix. The average values of production of natural (MN), dry matter (DM) and contents of Brix% were subjected to analysis of variance and regression for variables. The accumulation of dry matter and were influenced by fertilization. The contents of Brix% in two varieties did not suffer significant influence of fertilization.

Avaliação da influência da matéria orgânica, na determinação quantitativa de metais em amostras de sedimento, por espectrometria de absorção atômica e análise de metais traços por fluorescência de raios-X

The proposal of this work is to evaluate the influence of the organic matter on the results of the analyses of the metals (Zn, Pb, Al, Cu, Cr, Fe, Cd e Ni) for Atomic Absorption Spectrometry (AAS), so much in the extraction stage as in the reading using for that the chemometrics. They were used for this study sample of bottom sediment collected in river Jundiaí in the vicinity of the city of Macaíba-RN, commercial humus and water of the station of treatment of sewer of UFRN. Through the analyses accomplished by EAA it was verified that the interference of the organic matter happens in the extraction stage and not in the reading. With relationship to the technique of X Ray Fluorescence Spectrometry (XRFS), the present work has as intended to evaluate the viability of this technique for quantitative analysis of trace metals (Cr, Ni, Cu, Zn, Rb, Sr and Pb) in having leached obtained starting from the extraction with acqua regia for an aqueous solution. The used samples constitute the fine fraction (<0.063 mm) of sediments of swamp of the river Jundiaí. The preparation of tablets pressed starting from the dry residue of those leached it allowed your analysis in the solid form. This preliminary study shows that, in the case of the digestion chemistry partially of the fine fractions of bottom sediments used for environmental studies, the technique of applied EFRX to the analysis of dry residues starting from having leached with acqua regia, compared her it analyzes of the leached with ICP-OES, it presents relative mistakes for Cu, Pb, Sr and Zn below 10%