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Nowadays, the therapeutic efficiency of small interfering RNAs (siRNA) is still limited by the efficiency of gene therapy vectors capable of carrying them inside the target cells. In this study, siRNA nanocarriers based on low molecular weight chitosan grafted with increasing proportions (5 to 55%) of diisopropylethylamine (DIPEA) groups were developed, which allowed precise control of the degree of ionization of the polycations at pH 7.4. This approach made obtaining siRNA nanocarriers with small sizes (100-200 nm), positive surface charge and enhanced colloidal stability (up to 24 h) at physiological conditions of pH (7.4) and ionic strength (150 mmol L-1) possible. Moreover, the PEGylation improved the stability of the nanoparticles, which maintained their colloidal stability and nanometric sizes even in an albumin-containing medium. The chitosan-derivatives displayed non-cytotoxic effects in both fibroblasts (NIH/3T3) and macrophages (RAW 264.7) at high N/P ratios and polymer concentrations (up to 0.5 g L-1). Confocal microscopy showed a successful uptake of nanocarriers by RAW 264.7 macrophages and a promising ability to silence green fluorescent protein (GFP) in HeLa cells. These results were confirmed by a high level of tumor necrosis factor-α (TNFα) knockdown (higher than 60%) in LPS-stimulated macrophages treated with the siRNA-loaded nanoparticles even in the FBS-containing medium, findings that reveal a good correlation between the degree of ionization of the polycations and the physicochemical properties of nanocarriers. Overall, this study provides an approach to enhance siRNA condensation by chitosan-based carriers and highlights the potential of these nanocarriers for in vivo studies.

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BACKGROUND: Injuries to the articular cartilage of the knee often fail to heal properly due to the hypocellular and avascular nature of this tissue. Subsequent disability can limit participation in sports and decrease quality of life. Subchondral bone perforations are used for the treatment of small defects. Filling out the central portion in larger lesions becomes difficult, and scaffolds can be used as adjuvants, providing a matrix onto which the defect can be filled in completely. Also, autogenous cartilage grafts can be combined, acting as an inducer and improving healing quality, all in a single procedure. METHODS: This observational study evaluated the clinical and quality-of-life outcomes of patients with articular cartilage lesions of the knee undergoing repair via a microfracture technique combined with a synthetic scaffold and autogenous cartilage graft, with transosseous sutures and fibrin glue fixation, at 12 months of follow-up. Secondarily, it assessed whether combined procedures, previous surgical intervention, traumatic aetiology, lesion location, and age affect outcomes. The sample consisted of adult patients (age 18-66 years) with symptoms consistent with chondral or osteochondral lesions, isolated or multiple, ICRS grade III/IV, 2-12 cm2 in size. Patients with corrected angular deviations or instabilities were included. Those with BMI > 40 kg/m2, prior total or subtotal (> 30%) meniscectomy, second-look procedures, and follow-up < 6 months were excluded. Pain (VAS), physical activity (IKDC), osteoarthritis (WOMAC), and general quality of life (SF-36) were assessed. RESULTS: 64 procedures were included, comprising 60 patients. There was significant improvement (P < 0.05) in VAS score (5.92-2.37), IKDC score (33.44-56.33), and modified WOMAC score (53.26-75.93) after surgery. The SF-36 showed significant improvements in the physical and mental domains (30.49-40.23 and 46.43-49.84 respectively; both P < 0.05). CONCLUSIONS: Combination of microfractures, autogenous crushed cartilage graft, synthetic scaffold, and transosseous sutures with fibrin glue provides secure fixation for treatment of articular cartilage lesions of the knee. At 12-month follow-up, function had improved by 20 points on the IKDC and WOMAC, and quality of life, by 10 points on the SF-36. Age > 45 years had a negative impact on outcomes.

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Magnesium supplementation may be beneficial for cancer patients due to its action as a modulator of cell proliferation and metabolism and its anti-inflammatory effect. Tumor metabolism can influence the bioavailability and absorption of nutrients, leading to an increase in the individual's nutritional needs. In this work, the effects of supplementing different dosages of magnesium chloride in mice with solid Ehrlich's tumors were investigated by analyzing their hematological, inflammatory and anthropometric biomarkers. Three dosages of magnesium chloride (MgCl2) were administered for 28 consecutive days. Animal welfare was assessed according to the criteria stipulated by the National Center for the Replacement, Refinement, and Reduction of Animals in Research (NC3Rs). The inverted grid method was used to analyze muscle strength and fatigue. Difference in expression of the Tumor Necrosis Factor (TNF-α) and the Growth Transformation Factor (TGF-ß1) genes was determined by the 2-ΔCt method. The hematological evaluation consisted of the erythrogram, white blood cell and platelet counts were used for the hematological evaluation and treatment cytotoxicity. Difference in the expression of the TNF-α and TGF-ß genes showed that the group that received a high dose of magnesium had a decrease in TNF-α and RNL, an improvement in well-being with a tendency to increase muscle strength and less tumor progression according to the days of treatment. The group that received a low dosage of magnesium had a smaller tumor volume and a more controlled tumor growth according to the days. The group that received an intermediate dosage presented cytotoxicity.

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This study aims to contribute toward a better understanding of the current biodiversity patterns existing in the Atlantic Forest of the state of Rio de Janeiro from a paleo-ecological approach. Three soil profiles, each formed from distinct source materials under varied climate and vegetation conditions were selected from the coastal regions of Rio de Janeiro, Brazil. Soil horizon samples for chemical and physical characterization were collected from 10 cm-interval depths, and the total organic carbon was determined in addition to performing isotype and phytolithic analyses. The phytolithic analysis associated with the isotopic techniques (δ13C) permitted the characterization of three stages of paleo-environmental evolution for the studied soils, presenting valid trends that indicate small variations within the phytosociological structure of the vegetation along a Spodosols line in the studied areas. These stages indicated that the phytolithic assemblage was formed from a vegetation profile containing more trees, adapted to conditions colder than the existing vegetation, although no environmental changes were detected using the isotopic data. The milder temperature conditions may have favored the pedogenetic processes of the spodic B horizon formation as well as the maintenance of the organic matter contained in the soil. This study serves as the first draft for the paleo-environmental association among the regions where the analyzed profiles are found.

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We developed a label-free potentiometric biosensor using tyrosinase extracted from Musa acuminata and immobilized by covalent bond on a surface of a solid-contact transducer. The transducer was manufactured containing two layers. The first layer contained a blend of poly(vinyl) chloride carboxylated (PVC-COOH), graphite and potassium permanganate. On this layer, we deposited a second layer containing just a mixture of poly(vinyl chloride) carboxylated and graphite. On the last layer of the transducer, we immobilized the tyrosinase enzyme by reaction with N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride. The solid-contact potentiometric biosensor presented at low detection limit of 7.3×10-7M and a linear range to catechol concentration between 9.3×10-7M and 8.3×10-2M. This biosensor was applied to determine the amount of total phenols in different samples of honey and propolis. The results agreed with the Folin-Ciocalteu method.

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Changes in soil use and management can affect the soil aggregation, including aggregate formation by biogenic and physicogenic processes. The aim of this study was to analyze physical and biological influences on the genesis of soil aggregates in areas with different vegetations in the Atlantic Forest, as well as to compare physical and chemical attributes of the aggregates formed in different ways. Undeformed soil samples were collected from a depth of 0-10 cm from second-growth forests at different successional stages and a pasture area. To identify the pathways of aggregation we used morphological patterns proposed by Bullock et al. (1985), and established three clusters: physicogenic, biogenic, and intermediates. The aggregates were analyzed for stability, exchangeable cations, distribution of total organic carbon (TOC), and oxidizable fractions of total organic carbon. In all areas evaluated, the percentage of physicogenic aggregates was higher than that of biogenic and intermediate aggregates. The biogenic aggregates with the highest mean weight diameter (MWD) and geometric mean diameter (GMD) were recorded for samples from the second-growth forest at an advanced stage of succession (SFAS) and from the mixed managed pasture (MMP) sites (MWD: 4.520 mm and 4.896 mm; GMD: 3.678 mm and 4.479 mm, respectively). The biogenic aggregates presented higher levels of K and P compared to the other morphological types in all areas studied, with higher P levels in the SFAS area. The TOC content was also higher in the biogenic aggregates in all study areas, with 22.33 g kg-1 in SFAS, 25.60 g kg-1 in the site with second-growth forest at the middle stage (SFMS) of succession, 24.74 g kg-1 in the site with second-growth forest at the initial stage (SFIS) of succession, and 20.28 g kg-1 in MMP. The highest content of the fractions F1 (6.93 g kg-1) and F2 (7.43 g kg-1) were found in the biogenic class compared to the intermediate and physicogenic aggregates.[...](AU)

Mudanças no uso e manejo do solo podem afetar os processos de agregação, incluindo a formação de agregados por processos biogênicos e fisiogênicos. O objetivo deste estudo foi analisar a gênese de agregados por diferentes vias de formação, bem como atributos físicos e químicos dos agregados formados por essas vias em áreas com diferentes coberturas vegetais. Foram coletadas amostras de solo indeformadas na camada de 0-10 cm em áreas de floresta secundária com diferentes estádios sucessionais e uma área de pastagem. Para identificar as vias de agregação foram usados padrões morfológicos propostos por Bullock et al. (1985) e estabeleceu-se três grupos: fisiogênicos, biogênicas e intermediários. Os agregados foram analisados quanto à estabilidade em água, cátions trocáveis, teor de carbono orgânico total (COT) e frações oxidáveis do carbono orgânico total. Em todas as áreas avaliadas a porcentagem de agregados fisiogênicos foi maior do que a de agregados biogênicos e intermediários. Os agregados biogênicos foram encontrados em quantidade menor, com as maiores médias de Diâmetro Médio Ponderado (4.520 milímetros e 4.896 milímetros) e Diâmetro Médio Geométrico (3.678 milímetros e 4.479 milímetros) nas áreas de Floresta Secundária Estádio Avançado (FSEA) e Pasto Misto Manejado (PMM). Os agregados biogênicos apresentaram níveis mais elevados de K e P entre as classes morfológicas em todas as áreas estudadas, com os níveis de fósforo mais elevados na área de FSEA. O conteúdo COT também foi maior nos agregados biogênicos em todas as áreas de estudo, com 22.33 g kg-1 na FSEA, 25.60 g kg-1 na Floresta Secundária Estadio Médio (FSEM), 24.74 g kg-1 na Floresta Secundária Estadio Inicial (FSEI) e 20.28 g kg-1 em PMM. O maior teor de frações F1 (6.93 g kg-1) e F2 (7.43 g kg-1) foram encontrados na classe biogênica em comparação com agregados intermediários e fisiogênicos.[...](AU)

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Changes in soil use and management can affect the soil aggregation, including aggregate formation by biogenic and physicogenic processes. The aim of this study was to analyze physical and biological influences on the genesis of soil aggregates in areas with different vegetations in the Atlantic Forest, as well as to compare physical and chemical attributes of the aggregates formed in different ways. Undeformed soil samples were collected from a depth of 0-10 cm from second-growth forests at different successional stages and a pasture area. To identify the pathways of aggregation we used morphological patterns proposed by Bullock et al. (1985), and established three clusters: physicogenic, biogenic, and intermediates. The aggregates were analyzed for stability, exchangeable cations, distribution of total organic carbon (TOC), and oxidizable fractions of total organic carbon. In all areas evaluated, the percentage of physicogenic aggregates was higher than that of biogenic and intermediate aggregates. The biogenic aggregates with the highest mean weight diameter (MWD) and geometric mean diameter (GMD) were recorded for samples from the second-growth forest at an advanced stage of succession (SFAS) and from the mixed managed pasture (MMP) sites (MWD: 4.520 mm and 4.896 mm; GMD: 3.678 mm and 4.479 mm, respectively). The biogenic aggregates presented higher levels of K and P compared to the other morphological types in all areas studied, with higher P levels in the SFAS area. The TOC content was also higher in the biogenic aggregates in all study areas, with 22.33 g kg-1 in SFAS, 25.60 g kg-1 in the site with second-growth forest at the middle stage (SFMS) of succession, 24.74 g kg-1 in the site with second-growth forest at the initial stage (SFIS) of succession, and 20.28 g kg-1 in MMP. The highest content of the fractions F1 (6.93 g kg-1) and F2 (7.43 g kg-1) were found in the biogenic class compared to the intermediate and physicogenic aggregates.[...]

Mudanças no uso e manejo do solo podem afetar os processos de agregação, incluindo a formação de agregados por processos biogênicos e fisiogênicos. O objetivo deste estudo foi analisar a gênese de agregados por diferentes vias de formação, bem como atributos físicos e químicos dos agregados formados por essas vias em áreas com diferentes coberturas vegetais. Foram coletadas amostras de solo indeformadas na camada de 0-10 cm em áreas de floresta secundária com diferentes estádios sucessionais e uma área de pastagem. Para identificar as vias de agregação foram usados padrões morfológicos propostos por Bullock et al. (1985) e estabeleceu-se três grupos: fisiogênicos, biogênicas e intermediários. Os agregados foram analisados quanto à estabilidade em água, cátions trocáveis, teor de carbono orgânico total (COT) e frações oxidáveis do carbono orgânico total. Em todas as áreas avaliadas a porcentagem de agregados fisiogênicos foi maior do que a de agregados biogênicos e intermediários. Os agregados biogênicos foram encontrados em quantidade menor, com as maiores médias de Diâmetro Médio Ponderado (4.520 milímetros e 4.896 milímetros) e Diâmetro Médio Geométrico (3.678 milímetros e 4.479 milímetros) nas áreas de Floresta Secundária Estádio Avançado (FSEA) e Pasto Misto Manejado (PMM). Os agregados biogênicos apresentaram níveis mais elevados de K e P entre as classes morfológicas em todas as áreas estudadas, com os níveis de fósforo mais elevados na área de FSEA. O conteúdo COT também foi maior nos agregados biogênicos em todas as áreas de estudo, com 22.33 g kg-1 na FSEA, 25.60 g kg-1 na Floresta Secundária Estadio Médio (FSEM), 24.74 g kg-1 na Floresta Secundária Estadio Inicial (FSEI) e 20.28 g kg-1 em PMM. O maior teor de frações F1 (6.93 g kg-1) e F2 (7.43 g kg-1) foram encontrados na classe biogênica em comparação com agregados intermediários e fisiogênicos.[...]

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The purpose of this work was to improve the functional properties of chitosan for gene transfer by inserting phosphorylcholine (PC) and diethylaminoethyl (DEAE) groups into the main chain. A series of derivatives containing increasing contents of DEAE and a fixed content of PC groups were synthesized and characterized, aiming to evaluate the effect of these groups on the nanoparticles' properties and the in vitro transfection efficiency. The derivatives were soluble at physiological pH levels and all derivatives were less cytotoxic than the control, the lipid lipofectamine. The obtained derivatives complexed pDNA into nanoparticles with smaller sizes and higher zeta potentials than plain chitosan. The in vitro transfection was performed with nanoparticles prepared at pH 6.3 and 7.4 and the results showed that nanoparticles prepared with derivatives containing 20% of PC groups (PC18-CH) and high degrees of substitution by DEAE (PC20-CH-DEAE100, CH-DEAE80, CH-DEAE100) displayed the better transfection efficiencies in HeLa cells, reaching relative values comparable to lipofectamine. The most effective derivative, PC18CH, was selected for complexation with siRNA and its complexes demonstrated an in vitro knockdown efficiency highly dependent on the N/P ratio. Our combined results indicated that, by means of controlled modifications, the limitations of chitosan can be overcome to obtain more effective carriers based on chitosan, and the derivatives here studied hold potential for in vivo studies.

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Chitosan has been indicated as a safe and promising polycation vector for gene delivery. However its low transfection efficiency has been a challenging obstacle for its application. To address this limitation, we synthesized chitosan derivatives which had increasing amounts of diethylethylamine groups (DEAE) attached to the chitosan main chain. The plasmid DNA VR1412 (pDNA), encoding the ß-galactosidase (ß-gal) reporter gene was used to prepare nanoparticles with the chitosan derivatives, and the transfection studies were performed with HeLa cells. By means of dynamic light scattering and zeta potential measurements, it was shown that diethylethylamine-chitosan derivatives (DEAE(x)-CH) were able to condense DNA into small particles having a surface charge depending on the polymer/DNA ratio (N/P ratio). Nanoparticles prepared with derivatives containing 15 and 25% of DEAE groups (DEAE(15)-CH and DEAE(25)-CH) exhibited transfection efficiencies ten times higher than that observed with deacetylated chitosan (CH). For derivatives with higher degrees of substitution (DS), transfection efficiency decreased. The most effective carriers showed low cytotoxicity and good transfection activities at low charge ratios (N/P). Vectors with low DS were easily degraded in the presence of lysozyme at physiological conditions in vitro and the nontoxicity displayed by these vectors opens up new opportunities in the design of DEAE-chitosan-based nanoparticles for gene delivery.

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The interactions between phosphorylcholine-substituted chitosans (PC-CH) and calf-thymus DNA (ct-DNA) were investigated focusing on the effects of the charge ratio, the pH, and phosphorylcholine content on the size and stability of the complexes using the ethidium bromide fluorescence assay, gel electrophoresis, dynamic light scattering, and fluorescence microscopy. The size and colloidal stability of deacetylated chitosan (CH/DNA) and PC-CH/DNA complexes were strongly dependent on phosphorylcholine content, charge ratios, and pH. The interaction strengths were evaluated from ethidium bromide fluorescence, and, at N/P ratios higher than 5.0, no DNA release was observed in any synthesized PC-CH/DNA polyplexes by gel electrophoresis. The PC-CH/DNA polyplexes exhibited a higher resistance to aggregation compared to deacetylated chitosan (CH) at neutral pH. At low pH values highly charged chitosan and its phosphorylcholine derivatives had strong binding affinity with DNA, whereas at higher pH values CH formed large aggregates and only PC-CH derivatives were able to form small nanoparticles with hydrodynamic radii varying from 100 to 150 nm. Nanoparticles synthesized at low ionic strength with PC-CH derivatives containing moderate degrees of substitution (DS=20% and 40%) remained stable for weeks. Photomicroscopies also confirmed that rhodamine-labeled PC(40)CH derivative nanoparticles presented higher colloidal stability than those synthesized using deacetylated chitosan. Accordingly, due to their improved physicochemical properties these phosphorylcholine-modified chitosans provide new perspectives for controlling the properties of polyplexes.

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Os autores avaliaram 20 pacientes submetidos a tratamento cirúrgico por artrite séptica do quadril no período de novembro de 1964 a março de 1988 no Hospital de Clínicas da UFPR. A média de idade no momento do atendimento foi de 9,3 anos, com seguimento médio pós-operatório de 10,3 anos. Os resultados foram classificados segundo os critérios de Griffin & Green, dos pontos de vista clínico e radiográfico. A análise estatística mostrou que os melhores resultados foram obtidos nos casos em que o tratamento foi instituído assim que surgiram os sintomas e quando foi utilizada instilaçäo-aspiraçäo após a drenagem cirúrgica. Näo houve diferença estatisticamente significativa nos resultados dos nossos pacientes em relaçäo às diversas faixas etárias estudadas

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Os autores apresentam o caso de um paciente de 38 anos portador de mmesotelioma peritoneal cujas manifestaçöes clínicas de ascite recidivante e rebelde a terapêutica com diuréticos, dor abdominal, alteraçöes do comportamento e derrame pleural apontavam a evoluçäo clínica agressiva de tal patologia abdominal ainda sem terapêutica específica eficaz

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