Crystal structure of a soluble decoy receptor IL-22BP bound to interleukin-22

Interleukin-22 (IL-22) plays an important role in the regulation of immune and inflammatory responses in mammals. The IL-22 binding protein (IL-22BP), a soluble receptor that specifically binds IL-22, prevents the IL-22/interleukin-22 receptor 1 (IL-22R1)/interleukin-10 receptor 2 (IL-10R2) complex assembly and blocks IL-22 biological activity. Here we present the crystal structure of the IL-22/IL-22BP complex at 2.75 angstrom resolution. The structure reveals IL-22BP residues critical for IL-22 binding, which were confirmed by site-directed mutagenesis and functional studies. Comparison of IL-22/IL-22BP and IL-22/IL-22R1 crystal structures shows that both receptors display an overlapping IL-22 binding surface, which is consistent with the inhibitory role played by IL-22 binding protein.

On the localization of water-soluble porphyrins in micellar systems evaluated by static and time-resolved frequency-domain fluorescence techniques

Fluorescence quenching of meso-tetrakis-4-sulfonatophenyl (TPPS4) and meso-tetrakis-4-N-methylpyridil (TMPyP) porphyrins is studied in aqueous solution and upon addition of micelles of sodium dodecylsulfate (SDS), cetyltrimethylammonium chloride (CTAC), N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (HPS) and t-octylphenoxypolyethoxyethanol (Triton X-100). Potassium iodide (KI) was used as quencher. Steady-state Stern-Volmer plots were best fitted by a quadratic equation, including dynamic (K-D) and static (K-s) quenching. Ks was significantly smaller than K-D. Frequency-domain fluorescence lifetimes allowed estimating bimolecular quenching constants, k(q). At 25 degrees C, in aqueous solution, TMPyP shows k(q), values a factor of 2-3 higher than the diffusional limit. TPPS4 shows collisional quenching with pH dependent k(q) values. For TMPyP quenching results are consistent with reported binding constants: a significant reduction of quenching takes place for SDS, a moderate reduction is observed for H PS and almost no change is seen for Triton X-100. Similar data were obtained at 50 C. For CTAC-TPPS4 system an enhancement of quenching was observed as compared to pure buffer. This is probably associated to accumulation of iodide at the cationic micellar interface. The attraction between CTAC headgroups and 1(-), and repulsion between SDS and 1(-), enhances and reduces the fluorescence quenching, respectively, of porphyrins located at the micellar interface. The small quenching of TPPS4 in Triton X-100 is consistent with strong binding as reported in the literature. (C) 2008 Elsevier B.V. All rights reserved.

Synthesis of Phthalocyanines-ALA Conjugates: Water-Soluble Compounds with Low Aggregation

Syntheses of two water-soluble phthalocyanines (Pc) containing 5-aminolevulinic acid (ALA) linked to the core structure are described. These compounds were prepared by using original functionalizations, and they present remarkable structural and photophysical features, indicating that they could be applied to photodynamic therapy (PDT).

Water-soluble Cu, Fe, Mn and Zn species in nuts and seeds

An approach was developed to estimate molecular weight distribution of water-soluble Cu, Fe, Mn and Zn species in Brazil nut, cupuassu seed and coconut pulp by size exclusion chromatography (SEC) coupled on-line to ultra-violet (UV) and off-line to graphite furnace atomic absorption spectrometry (GF-AAS) detectors and matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS). SEC-UV analytical signals showed the prevalence of high molecular weight (HMW) species (79-1.7 kDa for Brazil nut, 50-1.7 kDa for coconut pulp, and 34-1.7 kDa for cupuassu seeds). The Brazil nut SEC-UV, GF-AAS and MALDI-TOF mass spectra gave confirmation of the association of the elements with water-soluble compounds. The elemental profiles were associated with fractions of compounds of molecular weight 1.2-16 kDa for Brazil nut, 1.7-13 kDa for coconut pulp, and 1.2-7.6 kDa for cupuassu seeds. (C) 2009 Elsevier Inc. All rights reserved.

Sequencing of Spodoptera frugiperda midgut trehalases and demonstration of secretion of soluble trehalase by midgut columnar cells

Both soluble (SfTre1) and membrane-bound (SfTre2) trehalases occur along the midgut of Spodoptera frugiperda larvae. Released SfTre2 was purified as a 67 kDa protein. Its K(m) (1.6 mM) and thermal stability (half life 10 min at 62 degrees C) are different from the previously isolated soluble trehalase (K(m) = 0.47 mM; 100% stable at 62 degrees C). Two cDNAs coding for S. frugiperda trehalases have been cloned using primers based on consensus sequences of trehalases and having as templates a cDNA library prepared from total polyA-containing RNA extracted from midguts. One cDNA codes for a trehalase that has a predicted transmembrane sequence and was defined as SfTre2. The other, after being cloned and expressed, results in a recombinant trehalase with a K(m) value and thermal stability like those of native soluble trehalase. This enzyme was defined as SfTre1 and, after it was used to generate antibodies, it was immunolocalized at the secretory vesicles and at the glycocalyx of columnar cells. Escherichia coli trehalase 3D structure and sequence alignment with SfTre1 support a proposal regarding the residue modulating the pKa value of the proton donor.

Association of soluble tumor necrosis factor receptors 1 and 2 with nephropathy, cardiovascular events, and total mortality in type 2 diabetes

AIMS/HYPOTHESIS: Soluble tumor necrosis factor receptors 1 and 2 (sTNFR1 and sTNFR2) contribute to experimental diabetic kidney disease, a condition with substantially increased cardiovascular risk when present in patients. Therefore, we aimed to explore the levels of sTNFRs, and their association with prevalent kidney disease, incident cardiovascular disease, and risk of mortality independently of baseline kidney function and microalbuminuria in a cohort of patients with type 2 diabetes. In pre-defined secondary analyses we also investigated whether the sTNFRs predict adverse outcome in the absence of diabetic kidney disease. METHODS: The CARDIPP study, a cohort study of 607 diabetes patients [mean age 61 years, 44 % women, 45 cardiovascular events (fatal/non-fatal myocardial infarction or stroke) and 44 deaths during follow-up (mean 7.6 years)] was used. RESULTS: Higher sTNFR1 and sTNFR2 were associated with higher odds of prevalent kidney disease [odd ratio (OR) per standard deviation (SD) increase 1.60, 95 % confidence interval (CI) 1.32-1.93, p < 0.001 and OR 1.54, 95 % CI 1.21-1.97, p = 0.001, respectively]. In Cox regression models adjusting for age, sex, glomerular filtration rate and urinary albumin/creatinine ratio, higher sTNFR1 and sTNFR2 predicted incident cardiovascular events [hazard ratio (HR) per SD increase, 1.66, 95 % CI 1.29-2.174, p < 0.001 and HR 1.47, 95 % CI 1.13-1.91, p = 0.004, respectively]. Results were similar in separate models with adjustments for inflammatory markers, HbA1c, or established cardiovascular risk factors, or when participants with diabetic kidney disease at baseline were excluded (p < 0.01 for all). Both sTNFRs were associated with mortality. CONCLUSIONS/INTERPRETATIONS: Higher circulating sTNFR1 and sTNFR2 are associated with diabetic kidney disease, and predicts incident cardiovascular disease and mortality independently of microalbuminuria and kidney function, even in those without kidney disease. Our findings support the clinical utility of sTNFRs as prognostic markers in type 2 diabetes.

Comparison between a soluble antigen-based ELISA and IFAT in detecting antibodies against Babesia canis in dogs

O presente trabalho estudou um ensaio imunoenzimático (ELISA) indireto para a detecção de anticorpos anti-Babesia canis no soro de cães, tendo a Reação de Imunofluorescência Indireta (RIFI), como teste de referência O antígeno utilizado no ELISA do presente estudo consistiu em uma preparação antigênica solúvel de merozoítas B. canis e as diluições ótimas do antígeno, soros e conjugado foram determinadas por titulação em bloco, utilizando soros de referência positivos e negativos. A preparação antigênica solúvel de B. canis ótima foi de 10 µg.mL-1, com soros de referência positivos e negativos em uma única diluição de 1:100, e conjugado a 1:4.000. Um total de 246 amostras séricas foram colhidas em cães, durante a campanha de vacinação anti-rábica em Jaboticabal, São Paulo, Brasil e a presença de anticorpos anti-B. canis foi avaliada pelo ELISA e RIFI. Nestas condições, a média de absorbância dos soros de referência negativos foi de 0,129 ± 0,025, resultando em um ponto de corte de 0,323 (Nível de ELISA 3) e a média da absorbância dos soros de referência positivos foi de 2,156 ± 1,187. As amostras com sorologia positiva para B. canis por ELISA e RIFI foram 67,89% (n = 167) e 59,35% (n = 146), respectivamente. Os resultados obtidos sugerem que o ELISA descrito revelou-se um teste sorológico eficaz no diagnóstico da babesiose canina.

The status of soil phosphate fractions and the ability of fungi to dissolve hardly soluble phosphates

Brazilian soils predominantly consist of iron and aluminum oxides and have a low phosphorus content. The present study was carried out in order to assess the status of phosphate fractions in pasture, forest and agricultural soils and the ability of soil fungi to solubilize iron and aluminum phosphates. The abundance of P fractions in the soils studied occurred in the following order: Fe-P > reductant-soluble Fe-P > occluded Fe-P > occluded Al-P > Al-P > Ca-P. of the 481 fungi isolated, 33 showed the ability to solubilize the inorganic phosphates in culture. of these, 14 were considered to be high or very high solubilizers based on a solubilization capacity > 1000 mu g PO43- ml(-1). Isolate F-111 was the only one that dissolved all the insoluble phosphates used. Nine isolates solubilized both Al-P and Ca-P, and four other isolates only solubilized Ca-P. The highest number of isolates with high solubilization capacity were detected in pasture soil, followed by tropical rain forest and forest patch soils. Pasture soil presented both the largest contents of insoluble phosphates and the largest number of fungal isolates with phosphate-solubilizing ability. The range and size of P fractions influenced the number of fungi and their ability to solubilize hardly soluble phosphates. (c) 2004 Elsevier B.V. All rights reserved.

Water-soluble nutrients in aerial plant parts of peanut and white oat as affected by lime and gypsum application

A calagem e a gessagem influenciam a disponibilidade e a absorção de nutrientes, podendo alterar a quantidade de ácidos orgânicos na parte aérea das culturas. Essas alterações, quantificadas por meio da análise de nutrientes solúveis em água, podem potencializar o efeito da correção do solo, garantindo a sustentabilidade do sistema de semeadura direta. Nesse sentido, objetivou-se avaliar o efeito da aplicação superficial de calcário e gesso no teor de nutientes solúveis em água nos resíduos vegetais das culturas de amendoim e aveia-branca. O experimento foi realizado em Latossolo Vermelho distroférrico em Botucatu (SP), nos anos agrícolas de 2004/2005 e 2005/2006. O delineamento experimental foi em blocos casualizados em esquema de parcelas subdivididas, com quatro repetições, em que as parcelas foram constituídas por doses de calcário, e as subparcelas, pela presença ou ausência da aplicação de gesso. Foi cultivado amendoim no verão e aveia-branca no inverno, em toda a área experimental. Na cultura do amendoim a aplicação de gesso elevou os teores de Ca solúvel em água apenas na primeira safra, em razão do curto período entre a aplicação do produto e a determinação dos teores de nutientes solúveis no extrato vegetal da cultura. A calagem realizada nas culturas de amendoim e aveia-branca elevou os teores solúveis de Ca, Mg, K e não alterou o teor de Cu e reduziu os teores de Zn, Mn e Fe nos dois anos de cultivo. Já a gessagem reduziu os valores da condutividade elétrica do amendoim (2004/2005 e 2005/2006) e da aveia-branca (2004/2005).

Chemical Modification of Botryosphaeran: Structural Characterization and Anticoagulant Activity of a Water-Soluble Sulfonated (1 -> 3)(1 -> 6)-beta-D-Glucan

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

Upregulation of soluble and membrane-bound human leukocyte antigen G expression is primarily observed in the milder histopathological stages of chronic hepatitis C virus infection

Chronic hepatitis C virus (HCV) infection is a worldwide health problem that may evolve to cirrhosis and hepatocellular carcinoma. Incompletely understood immune system mechanisms have been associated with impaired viral clearance. The nonclassical class I human leukocyte antigen G (HLA-G) molecule may downregulate immune system cell functions exhibiting well-recognized tolerogenic properties. HCV genotype was analyzed in chronic HCV-infected patients. Because HLA-G expression may be induced by certain viruses, we evaluated the presence of HLA-G in the liver microenvironment obtained from 89 biopsies of patients harboring chronic HCV infection and stratified according to clinical and histopathological features. Overall, data indicated that HCV genotype 1 was predominant, especially subgenotype 1a, with a prevalence of 87%. HLA-G expression was observed in 45(51%) liver specimens, and it was more frequent in milder stages of chronic hepatitis (67.4%) than in moderate (27.8%; p = 0.009) and severe (36.0%; p = 0.021) stages of the disease. Altogether, these results suggest that the expression of HLA-G in the context of HCV is a complex process modulated by many factors, which may contribute to an immunologic environment favoring viral persistence. However, because the milder forms predominantly expressed HLA-G, a protective role of this molecule may not be excluded. (C) 2012 American Society for Histocompatibility and Immunogenetics. Published by Elsevier B.V. All rights reserved.

Study on soluble heavy metals with preconcentration by using a new modified oligosilsesquioxane sorbent

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

Effect of soluble fiber on hypertriglyceridemia and immune profile in HIV-positive individuals undergoing highly active antiretroviral therapy

The advent of highly active antiretroviral therapy (HAART), since 1996, represented a profound impact on the natural history of HIV-infection by promoting important and sustainable viral replication suppression and increasing survival and quality of life among seropositive patients. Nonetheless, antiretroviral therapy has been observed to be accompanied by metabolic alterations such as dyslipidemia, especially hypertriglyceridemia, insulin resistance, hyperglycemia and lipodystrophy (body fat redistribution). Epidemiological studies have demonstrated a correlation between high triglyceride (TG) levels and higher incidence of coronary artery disease (CAD). Some investigators suggest dietary intervention as part of hyperlipidemia treatment, including an increase in soluble fiber intake (10-25g/day). Whereas some studies have demonstrated that both cholesterol and serum triglyceride levels decrease with the use of food fiber, others have shown just a serum triglyceride decrease, and others failed to observe any alteration in lipid metabolism. The purpose of this study was to assess the effect of soluble fiber (R) (partially hydrolyzed guar gum) supplementation on hypertriglyceridemia and immune profile in HIVpositive individuals on HAART. Nineteen HIV-positive individuals with hypertriglyceridemia (serum levels >= 150 to < 500mg/dl) were studied. of these individuals, 63.16% were males and 36.84% females, with mean age of 43.52 +/- 9.22 years. These individuals had been on the same HAART regimen for at least six months, had no change in therapy during the study and received 20g/day of soluble fiber for four months at pre-established times. Clinical-nutritional, biochemical (total proteins, albumin, globulin, total cholesterol, LDL-c, HDL-c, TG, TG/HDL-c and LDLc/HDL-c), hematimetric (hemoglobin, hematocrit and total lymphocytes), and immunologic (lymphocytes T CD4(+), T CD8(+); T CD4(+)/CD8(+) ratio, viral load, TNF-alpha and IL-6) parameters were assessed in all patients at three time points (M0: pretreatment, M1: 30 days, and M2: four months after intervention). Significance level was set at 5% for all data statistically analyzed. Serum TG and TG/HDL-c ratio reduction was observed at all time points, but statistical significance was found just at M0 and M2. The remaining biochemical, hematimetric and immunologic parameters (lymphocytes T CD4(+), T CD8(+); T CD4(+)/ CD8(+) ratio, and viral load) showed no significant difference at all times. Regarding serum cytokines, TNF-alpha and IL-6 significantly decreased between M0 and M2, and only IL-6 reduced between M1 and M2. The data collected show that dietary and anthropometric parameters remained unchanged excluding potential confounding factors related with the effect of fiber supplementation on serum TG, TNF-alpha and IL-6. Thus, soluble fiber (R) contributed to an important reduction in hypertriglyceridemia and in the serum levels of the proinflammatory cytokines TNF-alpha and IL-6 in HIV-seropositive individuals on HAART. In addition, soluble fiber (R) might have minimized the process of atherosclerosis in these individuals, given that elevated serum levels of TG, TNF-alpha and IL-6 have been associated with the development of these lesions.

Time-dependent alterations of soluble and cellular components in human milk

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

Hepatoprotective effect of water soluble extract of Coleus barbatus on cholestasis on young rats

OBJETIVO: Testar os efeitos do extrato aquoso de Coleus barbatus (EAB) na cirrose biliar secundária por obstrução das vias biliares extra-hepáticas em ratos jovens. MÉTODOS: Quarenta ratos Wistar machos com 21 dias de vida (P21), foram distribuídos em quatro grupos de 10 animais, submetidos a operação simulada ou dupla ligadura e ressecção do ducto biliar (S ou L) combinados EAB e a Água (B ou A). No P48, foi medido o tempo de sono com o pentobarbital (TS). No P49, foram submetidos a eutanásia para a determinação das atividades séricas do aspartato aminotransferase (AST) e da alanina aminotransferases (ALT); após a eutanásia foram avaliados o peso fresco do fígado (PFF) e, em cortes histológicos do fígado, a freqüência de mitoses (FM), o número de áreas de necrose (NN), a intensidade da fibrose (IF) e da proliferação ductal (IPD). Os efeitos da colestase, os do EAB e suas interações foram testados pela ANOVA com dois fatores, e as comparações múltiplas pareadas foram realizadas pelo teste de S.N.K ou teste de Wilcoxon. Também foi determinada a correlação linear de Pearson entre as variáveis histológicas duas a duas separadamente para os grupos LA e LD. O nível de significância estatística para os vários testes foi de p do erro alfa <0,05. RESULTADOS: A colestase aumentou significativamente o TS, a ALT, a AST, o PFF, a MI, o NN, a IF e a IPD. O EAB diminuiu o TS e a IM nos animais sem colestase (operação simulada). O EAB diminuiu o TS, a ALT, a AST, o PFF, a MI, o NN e IF na colestase. No grupo LA houve correlação positiva entre a IPD e a IF, correlação negativa entre a IPD e a FM e correlação negativa entre a IF a FM. No grupo LD houve correlação negativa entre o NN e a IPD. CONCLUSÕES: Na ausência de colestase o EAB encurta o tempo de sono e diminui a freqüência de mitoses. O EAB apresenta efeito hepatoprotetor no modelo de cirrose biliar secundária a obstrução biliar extra-hepática.