MicroRNA-221 silencing attenuates the degenerated phenotype of intervertebral disc cells.

The aim of this study was to investigate the role of an antichondrogenic factor, MIR221 (miR‐221), in intervertebral disc degeneration (IDD), and provide basic information for the development of a therapeutic strategy for the disc repair based on specific nucleic acid based drugs, such as miR‐221 silencing. We established a relatively quick protocol to minimize artifacts from extended in vitro culture, without selecting the different types of cells from intervertebral disc (IVD) or completely disrupting extracellular matrix (ECM), but by using the whole cell population with a part of resident ECM. During the de‐differentiation process miR‐221 expression significantly increased. We demonstrated the effectiveness of miR‐221 silencing in driving the cells towards chondrogenic lineage. AntagomiR‐221 treated cells showed in fact a significant increase of expression of typical chondrogenic markers including COL2A1, ACAN and SOX9, whose loss is associated with IDD. Moreover, antagomiR‐221 treatment restored FOXO3 expression and increased TRPS1 expression levels attenuating the severity grade of degeneration, and demonstrating in a context of tissue degeneration and inflammation not investigated before, that FOXO3 is target of miR‐221. Data of present study are promising in the definition of new molecules useful as potential intradiscal injectable biological agents

Níveis plasmáticos de vitamina A, carotenóides e proteína ligadora de retinol em crianças com infecções respiratórias agudas e doenças diarréicas

The present study was carried out in order to assess the plasma levels of vitamin A, carotenoids and retinol binding protein (RBP) of three-hundred and eleven children aged from seven months to eleven years, who had a history of upper respiratory infection (URI), pneumonia and diarrhoea. The children were resident in the urban area of the Municipality of S. Paulo, Brazil, and were seen at the pediatric service of the one school-hospital. The data show that plasma vitamin A (µg/dl) and RBP (mg/ dl) levels in the diarrhoea (15.2 µg/dl; 1.7 mg/dl) and pneumonia (15.2 µg/dl; 0.7 mg/dl) groups were lower (pPlanejou-se um estudo com o objetivo de se avaliar os níveis plasmáticos de vitamina A, carotenóides e proteína ligadora de retinol (RBP) em 311 crianças, de 7 meses a onze anos de idade, com história de infecções das vias aéreas superiores (IVAS), pneumonia e diarréia, residentes na área urbana da Cidade de São Paulo, Brasil, e atendidas no serviço de pediatria de um hospital-escola. As dosagens de vitamina A e carotenóides realizaram-se pelo método de Neeld-Pearson e o RBP pelo método de Mancini. Os níveis plasmáticos de vitamina A (µg/dl) e RBP (mg/dl) foram mais baixos (

Peptidomics characterization of allergenic and non-allergenic tropomyosin orthologs

Background The cleavage and the digestion patterns of allergenic proteins play a key role in allergenicity. The transformation of food proteins starts with their denaturation by the acidic environment of the stomach. However, protein denaturation is not enough to completely remove allergenic properties of a protein, but it is necessary a complete enzymatic digestion. Whether the enzymatic digestion is not efficient, the persistence of bigger peptides can occur and put the basis for the development of the sensitization process.The hypothesis of the current work takes into consideration the probability that shrimp tropomyosin (TM) is not fully digested or presents a digestion pattern that generates some peptides that can be immunogenic.Therefore, the work plan designed, aims to study the cleavage pattern of: purified chicken TM, recombinant chicken TM, purified TM of Penaeus monodon (Pen m 1), recombinant TM of Penaeus monodon (rPen m 1) and recombinant TM of Crangon crangon (rCrac c 1). Methods One mg of each ortholog has been processed through simulated mouth, gastric and intestinal digestion. The sample was frozen to block the digestion and, after this step, concentrated and cleaned through protein precipitation. The protein pellet was processed for peptidomic analysis through 1D Tricine gel electrophoresis, 2D Tricine gel electrophoresis and mass spectrometry. Results Simulated gastric digestion pattern of shrimp TM highlighted the presence of a resistant band at an average MW of 25 kDa that could be involved in the immunogenic process. Conclusions This innovative approach (peptidomics study through 1D-2D Tricine/MS) could represent a milestone for the study of digestion patterns of allergenic proteins or for the study of allergenic potential of novel foods

Preliminary Evidence for Cell Membrane Amelioration in Children with Cystic Fibrosis by 5-MTHF and Vitamin B12 Supplementation: A Single Arm Trial

Cystic fibrosis (CF) is one of the most common fatal autosomal recessive disorders in the Caucasian population caused by mutations of gene for the cystic fibrosis transmembrane conductance regulator (CFTR). New experimental therapeutic strategies for CF propose a diet supplementation to affect the plasma membrane fluidity and to modulate amplified inflammatory response. The objective of this study was to evaluate the efficacy of 5-methyltetrahydrofolate (5-MTHF) and vitamin B12 supplementation for ameliorating cell plasma membrane features in pediatric patients with cystic fibrosis.A single arm trial was conducted from April 2004 to March 2006 in an Italian CF care centre. 31 children with CF aged from 3 to 8 years old were enrolled. Exclusion criteria were diabetes, chronic infections of the airways and regular antibiotics intake. Children with CF were supplemented for 24 weeks with 5-methyltetrahydrofolate (5-MTHF, 7.5 mg /day) and vitamin B12 (0.5 mg/day). Red blood cells (RBCs) were used to investigate plasma membrane, since RBCs share lipid, protein composition and organization with other cell types. We evaluated RBCs membrane lipid composition, membrane protein oxidative damage, cation content, cation transport pathways, plasma and RBCs folate levels and plasma homocysteine levels at baseline and after 24 weeks of 5-MTHF and vitamin B12 supplementation. In CF children, 5-MTHF and vitamin B12 supplementation (i) increased plasma and RBC folate levels; (ii) decreased plasma homocysteine levels; (iii) modified RBC membrane phospholipid fatty acid composition; (iv) increased RBC K(+) content; (v) reduced RBC membrane oxidative damage and HSP70 membrane association.5-MTHF and vitamin B12 supplementation might ameliorate RBC membrane features of children with CF.ClinicalTrials.gov NCT00730509

Comparative Proteomic Analysis of Serum from Patients with Systemic Sclerosis and Sclerodermatous GVHD. Evidence of Defective Function of Factor H

BACKGROUND: Systemic sclerosis (SSc) is an autoimmune disease characterized by immunological and vascular abnormalities. Until now, the cause of SSc remains unclear. Sclerodermatous graft-versus-host disease (ScGVHD) is one of the most severe complications following bone marrow transplantation (BMT) for haematological disorders. Since the first cases, the similarity of ScGVHD to SSc has been reported. However, both diseases could have different etiopathogeneses. The objective of this study was to identify new serum biomarkers involved in SSc and ScGVHD. METHODOLOGY: Serum was obtained from patients with SSc and ScGVHD, patients without ScGVHD who received BMT for haematological disorders and healthy controls. Bi-dimensional electrophoresis (2D) was carried out to generate maps of serum proteins from patients and controls. The 2D maps underwent image analysis and differently expressed proteins were identified. Immuno-blot analysis and ELISA assay were used to validate the proteomic data. Hemolytic assay with sheep erythrocytes was performed to evaluate the capacity of Factor H (FH) to control complement activation on the cellular surface. FH binding to endothelial cells (ECs) was also analysed in order to assess possible dysfunctions of this protein. PRINCIPAL FINDINGS: Fourteen differentially expressed proteins were identified. We detected pneumococcal antibody cross-reacting with double stranded DNA in serum of all bone marrow transplanted patients with ScGVHD. We documented higher levels of FH in serum of SSc and ScGVHD patients compared healthy controls and increased sheep erythrocytes lysis after incubation with serum of diffuse SSc patients. In addition, we observed that FH binding to ECs was reduced when we used serum from these patients. CONCLUSIONS: The comparative proteomic analysis of serum from SSc and ScGVHD patients highlighted proteins involved in either promoting or maintaining an inflammatory state. We also found a defective function of Factor H, possibly associated with ECs damage

Aspects of the encephalitis epidemic caused by arbovirus in the Ribeira Valley, S. Paulo, Brazil, during 1975-1978

In 1975, 1976, and 1977, arbovirus caused an encephalitis epidemic in the Ribeira Valley in the state of S. Paulo. The epidemic would peak when the temperature and pluvial levels were higher. From 1978 on the disease maintained low levels within a presumed endemic zone. The epidemic had swept from east to west and from east to southwest in a wave toward the neighboring coastal region. The mountain chain to the north and northwest acted as a barrier. It was only logical natural that the hypothesis that the etiological agent, arbovirus Rocio may have recently infected the human population be considered. Mosquitos must have transmitted the infection from birds and small mammals in nearby forests. Probable forms of transmission of arboviroses in the home are also discussed in this article as well as the fact that population groups that presented the worst forms of the disease were the very young, and the very old and those living in the worst conditions. The epidemiological perspective of the arboviroses is that it persists in this area because the area presents excellent conditions for the development of the etiological agent - reservoirs and biological vectors, with a continuous supply of susceptible people, migrants or tourists.Foi realizado estudo epidemiológico da encefalite por arbovirus na região do Vale do Ribeira, S. Paulo, Brasil. Uma epidemia da moléstia ocorreu em 1975, 1976 e 1977, com picos nas épocas de maior temperatura e pluviosidade. A partir de 1978 a moléstia manteve-se em níveis baixos numa presumível ende-micidade. A epidemia se deslocou em onda em direção leste-oeste e leste-sudoeste para a região litorânea vizinha. A cadeia montanhosa situada ao norte e noroeste atuou como barreira à propagação da moléstia. Considerou-se a hipótese que o agente etiológico, arbovirus Rocio, deva ter começado a infectar a população humana recentemente, tendo sido veiculado ao homem de reservatórios silvestres, aves e pequenos mamíferos, por culicídeos silvestres. Discutiu-se também prováveis formas de transmissão domiciliar. Verificou-se que os grupos populacionais que apresentaram as formas mais graves foram os de idades extremas e os que apresentavam piores condições de vida. Considerou-se que a perspectiva epidemiológica desta arbovirose é que ela persista na região, uma vez que a mesma apresenta condições ótimas para o desenvolvimento do agente etiológico, dos reservatórios e dos vetores biológicos, além de receber um contínuo afluxo de população suscetível, migrantes ou turistas

Proteomica

Questo volume è frutto di un’iniziativa emersa dall’esigenza di molti docenti, e qui Autori, che necessitavano di un libro completo di proteomica da adottare in diversi corsi universitari. Il testo è diviso in sezioni, che potrebbero corrispondere a moduli di corso: una parte più generale, una sezione che presenta i diversi metodi, una sezione molto più approfondita ed, infine, una parte finale più specifica, che si occupa di particolari procedure e della rielaborazione dei risultati proteomici. Il docente potrà quindi scegliere se adottarle tutte o inserirle in diversi corsi, a seconda delle proprie necessità e di quelle dei suoi studenti

Relação entre diarréia aguda e baixos níveis plasmáticos de vitamina A e proteína carregadora de retinol (RBP)

The objective of this study was to assess vitamin A status and association between acute diarrhoea and plasma levels of vitamin A through cross-sectional comparison in children. Plasma vitamin A was measured by colorimetric method of Neeld & Pearson and RBP by radial immunodiffusion technique. Seventy eight children (aged 18-119 months), 26 with current history of diarrhoea and 52 children as controls (outpatient from the Santa Casa de Misericórdia Hospital in metropolitan area of São Paulo City, Brazil) were studied. Children with history of diarrhoea showed significant low levels (mean &plusmn; s.e.) as compared to controls, vitamin A (15.87 &plusmn; 1.4 µg/dl vs. 21.14 &plusmn; 1.15 µg/dl, p < 0.007) and RBP (1.70 &plusmn; 0.2 mg/dl vs. 2.52 &plusmn;0.11 mg/dl). Multivariate logistic regression adjusted by sex, age, nutritional status and mother education revealed association between diarrhoea and inadequate levels of vitamin A and RBP.O objetivo deste trabalho foi avaliar o estado nutricional relativo a vitamina A e a associação entre diarréia e os níveis plasmáticos de vitamina A e RBP através de um estudo transversal. A vitamina A foi medida pelo método de Neeld & Pearson e RBP pela técnica de imunodifusão radial. Foram estudadas 78 crianças (com idades entre 18 e 119 meses), 26 com diarréia e 52 como controles, atendidas na consulta ambulatorial do Hospital Santa Casa de Misericórdia de São Paulo, da Cidade de São Paulo. As crianças com diarréia mostraram níveis significativamente mais baixos de vitamina A (15.87 &plusmn; 1.4 µg/dl vs. 21.14 &plusmn; 1.15 µg/dl, p< 0.007) e RBP (1.70 &plusmn; 0.2 mg/dl v.s 2.52 &plusmn; 0.11 mg/dl) quando comparadas com as do grupo controle. As análises de regressão logística mostraram associação entre diarréia e níveis inadequados de vitamina A e RBP ajustados pelo sexo, idade, estado nutricional e nível de educação materna