Uncaria tomentosa (UT) extracts have been shown to have promising anti-tumor activity. We hypothesized that its incorporation into nanostructured systems could improve the anticancer properties. ...Here, poly-e-caprolactone (PCL) and poly-d,l-lactide-co-glycolide (PLGA) were employed to generate nanoparticles loaded with UT extract in a single emulsion solvent evaporation method. The nanoparticles were characterized by particle size, zeta potential, morphology and entrapment efficiency along with stability and release profiles. The nanoparticles presented entrapment efficiencies above 60% and a mean diameter below 300nm. UT-PCL nanoparticles presented higher entrapment efficiency and mean particle size as well as a slow release rate. The UT-PLGA nanoparticles showed higher drug loading. Two prostate cancer cell-lines, LNCaP and DU145 that were derived from metastatic sites, served as model systems to assess cytotoxicity and anti-cancer activity. In vitro, both formulations reduced the viability of DU145 and LNCaP cells. Yet, the UT-PLGA nanoparticles showed higher cytotoxicity towards DU145 cells while the UTPCL against LNCaP cells. The results confirm that the incorporation of UT into nanoparticles could enhance its anti-cancer activities that can offer a viable alternative for the treatment of prostrate canner and highlights the potential of nanostructured systems to provide a promising methodology to enhance the activity of natural extracts.
The concern about the risks of viral infections transmission through blood transfusion has led into a search for improvements on screening tests used for the selection of blood donors. Molecular ...biology techniques applied in researches of viral genomes, known as Nucleic Acid-amplification-Test (NAT), represent a technology capable of increasing transfusion safety by shortening the diagnostic window period. In Brazil, the implementation of this technology for the detection of HIV, HCV and HBV occurred due to the implantation of the NAT Kit - produced by Immunobiological Technology Institute (Biomanguinhos-FIOCRUZ), in the Brazilian blood centers. The National Health Surveillance Agency attaches great importance to validation, since it standardizes, disciplines and regulates criteria for the registration of health products. This work aims to establish a protocol of performance validation by real-time PCR method, taking as the object of study the Bio-Manguinhos NAT Kit, in order to update the product registration or to meet any future needs to ensure all regulatory requirements for the performance validation of the real-time PCR diagnostic kit. The protocol developed followed the ICH recommendations. The results revealed that the adopted methodology contemplates the necessary requirements for compliance with the Brazilian legislation, as well as the established validation parameters.
Dermorphin is a peptide with analgesic actions similar to morphine, but with greater effect and less potential to cause tolerance. The use of dermorphin has been documented in race horses, and its ...use in humans has already been reported. Considering the potential advantages from the use of dermorphin over morphine, a method to monitor it, and its main metabolite dermorphin (1‐4) in humans becomes necessary for doping control. Here, we present two orthogonal methods for this purpose: a high‐throughput liquid chromatography coupled to high‐resolution mass spectrometry (HRMS) as an initial testing procedure and liquid chromatography–tandem mass spectrometry (MS/MS) in the selected reaction monitoring (SRM) acquisition mode for a confirmation procedure. For urine samples, pretreatment through a mixed‐mode weak cation‐exchange solid‐phase extraction emerged as an effective approach to extract peptides from the biological sample. For the HRMS analysis, a full‐MS scan acquisition mode was selected to detect the exact masses of dermorphin and dermorphin (1‐4) at m/z 803.37226 and 457.20816, respectively. The SRM method used in the MS/MS confirmation protocol presented high specificity and sensitivity. The selected product ions for dermorphin were 602.2, 202.1 and 574.3 and for dermorphin (1‐4) were 207.1, 223.1, and 235.1. Both methods were evaluated for specificity, repeatability, carryover, matrix effects, and recovery. No carryover and matrix effects were detected. The limit of detection for initial testing procedure and the limit of identification for confirmation procedure was 2.5 ng/ml. Also, specificity and robustness were acceptable for the application. Together, the developed methods proved to be efficient for the analysis of dermorphin and metabolite for human doping control purpose.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
Layered drug delivery carriers are current targets of nanotechnology studies since they are able to accommodate pharmacologically active substances and are effective at modulating drug release. ...Sodium montmorillonite (Na-MMT) is a clay that has suitable properties for developing new pharmaceutical materials due to its high degree of surface area and high capacity for cation exchange. Therefore Na-MMT is a versatile material for the preparation of new drug delivery systems, especially for slow release of protonable drugs. Herein, we describe the intercalation of several amine-containing drugs with Na-MMT so we can derive a better understanding of how these drugs molecules interact with and distribute throughout the Na-MMT interlayer space. Therefore, for this purpose nine sodium montmorillonite/amine-containing drugs complexes (Na-MMT/drug) were prepared and characterized. In addition, the physicochemical properties of the drugs molecules in combination with different experimental conditions were assessed to determine how these factors influenced experimental outcomes (e.g. increase of the interlayer spacing versus drugs arrangement and orientation). We also performed a molecular modeling study of these amine-containing drugs associated with different Na-MMT/drug complex models to analyze the orientation and arrangement of the drugs molecules in the complexes studied. Six amine-containing drugs (rivastigmine, doxazosin, 5-fluorouracil, chlorhexidine, dapsone, nystatin) were found to successfully intercalate Na-MMT. These findings provide important insights on the interlayer aspect of the molecular systems formed and may contribute to produce more efficient drug delivery nanosystems.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Dermorphin is a heptapeptide with an analgesic potential higher than morphine that does not present the same risk for the development of tolerance. These pharmacological features make dermorphin a ...potential doping agent in competitive sports and it is already prohibited for racehorses. For athletes, the development of an efficient strategy to monitor for its abuse necessitates an investigation of the metabolism of dermorphin in humans.
Here, human liver microsomes and zebrafish were utilized as model systems of human metabolism to evaluate the presence and kinetics of metabolites derived from dermorphin. Five hours after its administration, the presence of dermorphin metabolites could be detected in both models by liquid chromatography coupled to highresolution mass spectrometry.
Although the two models showed common results, marked differences were also observed in relation to the formed metabolites. Six putative metabolites, based on their exact masses of m/z 479.1915, m/z 501.1733, m/z 495.1657, m/z 223.1073, m/z 180.1017 and m/z 457.2085, are proposed to represent the metabolic pattern of dermorphin. The major metabolite generated from the administration of dermorphin in both models was YAFG-OH (m/z 457.2085), which is the N-terminal tetrapeptide previously identified from studies on rats.
Its extensive characterization and commercial availability suggest that it could serve as a primary target analyte for the detection of dermorphin misuse. The metabolomics approach also allowed the assignment of other confirmatory metabolites.
Abstract Sildenafil is a potent selective inhibitor of phosphosdiesterase-5 previously used in erectile dysfunction and subsequently approved in 2005 for pulmonary arterial hypertension treatment. ...Since oral administration of sildenafil shows pharmacokinetic problems with mean absolute bioavailability of 41%, the goal of this work was to develop a novel sildenafil self-emulsifying drug delivery system (SEDDS) for oral absorption improvement and management of dosage. One pharmaceutical solution and four SEDDS containing sildenafil were successfully obtained and SEDDS formed O/W nanoemulsion with droplet size less than 300 nm. The stability studies evidenced that the SEDDS containing 3.3% w/w of sildenafil yielded the best results. The safety of 2-pyrrolidone/isobutanol in oral formulations was assessed in mice and no lethality was achieved in the placebo groups with LD50 of 490 mg/Kg for SEDDS II-3.3, suggesting it as a safe excipient for humans. Therewithal, in silico studies using PBPK models provided the pharmacokinetic profile of sildenafil SEDDS. Subsequently, in silico evaluation indicated that the sildenafil SEDDS droplet size influenced its bioavailability, enhancing absorption, assuring a good pharmacokinetic profile. These findings suggest that the formulations developed here presented the potential to enhance drug oral absorption with the possibility to control drug dosage as they are liquid pharmaceutical formulations.
Dietary supplements in many countries such as the USA do not require registration prior to commercialization. The Agência Nacional de Vigilância Sanitária (ANVISA) registers substances with ...functional properties as foods. Lutein is a carotenoid with antioxidant activity available on the market. However, no regulatory mandates exist to govern the design of quality control tests, which are necessary to ensure formulation effectiveness. Therefore, in the present study, tablet and dosage formulations from different manufacturers were tested following general methods outlined in the Brazilian and American Pharmacopeias. The averageweight, disintegration, content and dose uniformity assays were performed for all tablets and capsules, whereas hardness assays were only performed on tablets. None of the 10 formulations studied were found to be of satisfactory quality. Of all tablets tested, two had no-significant available lutein content, which may indicate adulteration. The capsules displayed adequate amounts of lutein, however had alarmingly negative disintegration and dissolution test results, which may contribute to non-bioavailability of lutein. All formulations analyzed are currently being marketed in the Brazilian and American markets. The low physicochemical performance in these formulations can be explained by the lack of specific regulations, which are necessary to ensure the quality of lutein-containing products on the market.
The treatment of Diabetes Mellitus (DM), a chronic disease, is primarily based upon administration of insulin forms to patients. Conventional subcutaneous administration is associated with a large ...number of complications, therefore, several new strategies have been developed. Amongst these strategies, oral insulin administration is much less invasive and, therefore, well tolerated. In recent years, various nanoformulations were developed for the oral administration of insulin, allowing more effective stabilization of the active pharmaceutical ingredient and modified for better absorption along the gastrointestinal tract. The development of different oral insulin nanoformulations in academic research as well as in patents, including the development of nanoparticles, liposomes, nanoemulsions and the use of cyclodextrins deserves special attention. The future of oral insulin nanoformulations is dependent on strategies utilizing simple technologies that stabilize the raw material, including inclusion within cyclodextrins or inclusion in low weight molecular mass polymers/ oligomers. All of the theories developed here provide a solid foundation upon which to develop new methods for the production of pharmaceutical peptide formulations. In addition, the effective search for existing nanometric formulations of insulin could provide economically viable therapeutic options that can consequently be produced on an industrial scale.
ABSTRACT Considering the cashew tree’s relevance and the limitations imposed by salinity stress in semi-arid regions, the use of alternatives capable of mitigating the harmful effects due to salinity ...is of great importance to the production sector. The use of grafted plants, especially with rootstock made of tolerant materials, influences the accumulation of toxic ions in leaves of grafted seedlings. Thus, the objective of this work was to evaluate morphophysiological characteristics and leaf concentrations of Na+, K+ and Ca+2 of combinations of scion and rootstock of early dwarf cashew, contrasting in terms of salinity tolerance. The experiment was carried out in a completely randomized design with five replicates, in a 4 × 3 factorial arrangement, corresponding to four dwarf cashew scion/rootstock combinations (self-graft CCP 09, CCP 09/CCP 76, self-graft CCP 76, and CCP 76/CCP 09) and three NaCl concentrations (0, 50, and 100 mM L-1). Height, number of leaves, leaf area, dry matter, tolerance index and leaf concentrations of Na+, K+ and Ca+2 were evaluated after 30 days of application of NaCl concentrations. The scion/rootstock combination CCP 76/09 showed tolerance to 50 mM L-1, due to the increase of leaf area and number of leaves. The scion/rootstock combination CCP 76/09 was more suitable, as it kept the leaf K+ concentration and had the lowest Na+ concentration.
RESUMO Considerando a relevância do cajueiro e a limitação imposta pelo estresse salino em regiões semiáridas, o uso de alternativas capazes de mitigar os efeitos deletérios em decorrência da salinidade são de grande importância ao setor produtivo. O uso de plantas enxertadas, principalmente com porta-enxertos de materiais tolerantes, influencia no acúmulo de íons tóxicos nas folhas das mudas enxertadas. Assim, o objetivo deste trabalho foi avaliar características morfofisiológicas e concentração foliar de Na+, K+, Ca+2 de combinações de copa e porta-enxerto de cajueiro anão precoce, contrastantes à salinidade. O experimento foi realizado em delineamento inteiramente casualizado, com cinco repetições, tendo os tratamentos dispostos em arranjo fatorial de 4 × 3, correspondentes a quatro combinações de mudas enxertadas (auto-enxerto CCP 09/CCP 09, CCP 09/CCP 76, auto-enxerto CCP 76/CCP 76 e CCP 76/CCP 09) e a três concentrações de NaCl (0-controle, 50 e 100 mM L-1). Foram avaliados altura, número de folhas, área foliar, massa seca, índice de tolerância e concentração foliar de Na+, K+ e Ca+2 após 30 dias da aplicação das concentrações de NaCl. A combinação copa/porta-enxerto CCP 76/09 apresentou tolerância à salinidade de 50 mM L-1, devido ao aumento da área foliar e número de folhas. A combinação copa/porta-enxerto CCP 76/09 foi mais adequada, pois manteve a concentração foliar de K+ e o menor concentração de Na+.
The litterfall is the major organic material deposited in soil of Brazilian Caatinga biome, thus providing the ideal conditions for plant biomass-degrading microorganisms to thrive. Herein, the ...phylogenetic composition and lignocellulose-degrading capacity have been explored for the first time from a fosmid library dataset of Caatinga soil by sequence-based screening. A complex bacterial community dominated by Proteobacteria and Actinobacteria was unraveled. SEED subsystems-based annotations revealed a broad range of genes assigned to carbohydrate and aromatic compounds metabolism, indicating microbial ability to utilize plant-derived material. CAZy-based annotation identified 7275 genes encoding 37 glycoside hydrolases (GHs) families related to hydrolysis of cellulose, hemicellulose, oligosaccharides and other lignin-modifying enzymes. Taxonomic affiliation of genes showed high genetic potential of the phylum Acidobacteria for hemicellulose degradation, whereas Actinobacteria members appear to play an important role in celullose hydrolysis. Additionally, comparative analyses revealed greater GHs profile similarity among soils as compared to the digestive tract of animals capable of digesting plant biomass, particularly in the hemicellulases content. Combined results suggest a complex synergistic interaction of community members required for biomass degradation into fermentable sugars. This large repertoire of lignocellulolytic enzymes opens perspectives for mining potential candidates of biochemical catalysts for biofuels production from renewable resources and other environmental applications.
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