Liposomal Delivery of Saquinavir to Macrophages Overcomes Cathepsin Blockade by Mycobacterium tuberculosis and Helps Control the Phagosomal Replicative Niches.

Mycobacterium tuberculosis is able to establish a chronic colonization of lung macrophages in a controlled replication manner, giving rise to a so-called latent infection. Conversely, when intracellular bacteria undergo actively uncontrolled replication rates, they provide the switch for the active infection called tuberculosis to occur. Our group found that the pathogen is able to manipulate the activity of endolysosomal enzymes, cathepsins, directly at the level of gene expression or indirectly by regulating their natural inhibitors, cystatins. To provide evidence for the crucial role of cathepsin manipulation for the success of tuberculosis bacilli in their intracellular survival, we used liposomal delivery of saquinavir. This protease inhibitor was previously found to be able to increase cathepsin proteolytic activity, overcoming the pathogen induced blockade. In this study, we demonstrate that incorporation in liposomes was able to increase the efficiency of saquinavir internalization in macrophages, reducing cytotoxicity at higher concentrations. Consequently, our results show a significant impact on the intracellular killing not only to reference and clinical strains susceptible to current antibiotic therapy but also to multidrug- and extensively drug-resistant (XDR) Mtb strains. Altogether, this indicates the manipulation of cathepsins as a fine-tuning strategy used by the pathogen to survive and replicate in host cells.

Innovative, Sugar-Free Oral Hydrogel as a Co-administrative Vehicle for Pediatrics: a Strategy to Enhance Patient Compliance.

Palatability and swallowability in the pediatric population are perceived as true challenges in the oral administration of medication. Pediatric patients have high sensitivity to taste and reduced ability to take solid dosage forms, which can often lead to a poor therapeutic compliance. It is crucial to find new strategies to simplify the oral administration of drugs to children. The present paper reports the development of a new hydrogel vehicle adapted to the pediatric population. Several polymers with similar properties were selected and adjustments were made to obtain the desired characteristics of the final product. The developed formulations were studied for organoleptic properties, rheology, mucoadhesion properties, preservative efficacy, and stability. Physical and chemical compatibilities between the vehicle and several drugs/medicines, at the time of administration, were also studied. Six final formulations with different polymers, odor, and color were chosen, and no known interactions with medications were observed. The proposed new oral vehicles are the first sugar-free vehicle hydrogels designed to make the intake of oral solid forms a more pleasant and safer experience for pediatric patients.

Repurposing of Marine Raw Materials in the Formulation of Innovative Plant Protection Products.

Over the years, the growth of the world population has caused a huge agricultural production to support the population's needs. Since plant protection products are essential to preserve agricultural crops and to optimize vital plant processes, it is crucial to use more sustainable, biodegradable, and biocompatible raw materials, without harming the environment and human health. Although the development of new plant protection products is a costly process, the environmental benefits should be considered. In this context, marine raw materials obtained as byproducts of fishing industries, possessing a wide variety of physicochemical and biological properties, can serve as a promising source of such materials. They have a high potential for developing alternative and safe formulations for agricultural applications, not only as biocompatible excipients but also as effective and selective, or even both. It is also possible to promote a synergistic effect between an active substance and the biological activity of the marine polymer used in the formulation, enabling plant protection products with lower concentrations of the active substances. Thus, this review addresses the repurposing of marine raw materials for the development of innovative plant protection products, focusing on micro- and nanoparticulate formulations, to protect the environment through more ecological and sustainable strategies.

A Newfangled Collagenase Inhibitor Topical Formulation Based on Ethosomes with Sambucus nigra L. Extract.

Sambucus nigra L. (S. nigra) is a shrub widespread in Europe and western Asia, traditionally used in medicine, that has become popular in recent years as a potential source of a wide range of interesting bioactive compounds. The aim of the present work was to develop a topical S. nigra extract formulation based on ethosomes and thus to support its health claims with scientific evidence. S. nigra extract was prepared by an ultrasound-assisted method and then included in ethosomes. The ethosomes were analyzed in terms of their size, stability over time, morphology, entrapment capacity (EC), extract release profile, stability over time and several biological activities. The prepared ethosomes were indicated to be well defined, presenting sizes around 600 nm. The extract entrapment capacity in ethosomes was 73.9 ± 24.8%, with an interesting slow extract release profile over 24 h. The extract-loaded ethosomes presented collagenase inhibition activity and a very good skin compatibility after human application. This study demonstrates the potential use of S. nigra extract incorporated in ethosomes as a potential cosmeceutical ingredient and on further studies should be performed to better understand the impact of S. nigra compounds on skin care over the time.

Sorting hidden patterns in nanoparticle performance for glioblastoma using machine learning algorithms.

Cationic compounds have been described to readily penetrate cell membranes. Assigning positive charge to nanosystems, e.g. lipid nanoparticles, has been identified as a key feature to promote electrostatic binding and design ligand-based constructs for tumour targeting. However, their intrinsic high cytotoxicity has hampered their biomedical application. This paper seeks to establish which cationic compounds and properties are compelling for interface modulation, in order to improve the design of tumour targeted nanoparticles against glioblastoma. How can intrinsic features (e.g. nature, structure, conformation) shape efficacy outcomes? In the quest for safer alternative cationic compounds, we evaluate the effects of two novel glycerol-based lipids, GLY1 and GLY2, on the architecture and performance of nanostructured lipid carriers (NLCs). These two molecules, composed of two alkylated chains and a glycerol backbone, differ only in their polar head and proved to be efficient in reversing the zeta potential of the nanosystems to positive values. The use of unsupervised and supervised machine learning (ML) techniques unraveled their structural similarities: in spite of their common backbone, GLY1 exhibited a better performance in increasing zeta potential and cytotoxicity, while decreasing particle size. Furthermore, NLCs containing GLY1 showed a favorable hemocompatible profile, as well as an improved uptake by tumour cells. Summing-up, GLY1 circumvents the intrinsic cytotoxicity of a common surfactant, CTAB, is effective at increasing glioblastoma uptake, and exhibits encouraging anticancer activity. Moreover, the use of ML is strongly incited for formulation design and optimization.

Further Evidence of Possible Therapeutic Uses of Sambucus nigra L. Extracts by the Assessment of the In Vitro and In Vivo Anti-Inflammatory Properties of Its PLGA and PCL-Based Nanoformulations.

Sambucus nigra L. is widely used in traditional medicine with different applications. However, confirmative studies are strongly required. This study aimed to assess the biological activities of the S. nigra flower's extract encapsulated into two different types of nanoparticles for optimizing its properties and producing further evidence of its potential therapeutic uses. Different nanoparticles (poly(lactide-co-glycolide, PLGA) and poly-Ɛ-caprolactone (PCL), both with oleic acid, were prepared by emulsification/solvent diffusion and solvent-displacement methods, respectively. Oleic acid was used as a capping agent. After the nanoparticles' preparation, they were characterized and the biological activities were studied in terms of collagenase, in vitro and in vivo anti-inflammatory, and in vitro cell viability. Rutin and naringenin were found to be the major phenolic compounds in the studied extract. The encapsulation efficiency was higher than 76% and revealed to have an impact on the release of the extract, mainly for the PLGA. Moreover, biochemical and histopathological analyses confirmed that the extract-loaded PLGA-based nanoparticles displayed the highest anti-inflammatory activity. In addition to supporting the previously reported evidence of potential therapeutic uses of S. nigra, these results could draw the pharmaceutical industry's interest to the novelty of the nanoproducts.

Solid Lipid Nanoparticles and Nanostructured Lipid Carriers as Smart Drug Delivery Systems in the Treatment of Glioblastoma Multiforme.

Glioblastoma multiforme (GBM) is the most common and malignant type of brain tumor. In fact, tumor recurrence usually appears a few months after surgical resection and chemotherapy, mainly due to many factors that make GBM treatment a real challenge, such as tumor location, heterogeneity, presence of the blood-brain barrier (BBB), and others. Solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs) represent the most promising carriers for therapeutics delivery into the central nervous system (CNS) owing to their inherent ability to cross the BBB. In this review, we present the main challenges in GBM treatment, a description of SLNs and NLCs and their valuable role as drug carriers in GBM treatment, and finally, a detailed description of all modification strategies that aim to change composition of SLNs and NLCs to enhance treatment outcomes. This includes modification of SLNs and NLCs to improve crossing the BBB, reduced GBM cell resistance, target GBM cells selectively minimizing side effects, and modification strategies to enhance SLNs and NLCs nose-to-brain delivery. Finally, future perspectives on their use are also be discussed, to provide insight about all strategies with SLNs and NLCs formulation that could result in drug delivery systems for GBM treatment with highly effective theraputic and minimum undesirable effects.

New nanoparticles for topical ocular delivery of erythropoietin.

Erythropoietin (EPO) is known for its neuroprotective and neuroregenerative properties. EPO topical ocular administration has not been tested yet and its bioavailability could be improved by mucoadhesive hydrogels. Thus, this study aimed to develop and evaluate a chitosan (CS) and hyaluronic acid (HA) nanoparticulate system for topical ocular delivery of EPO. Nanoparticles were prepared by ionotropic gelation using six different HAs (HA1-HA6), and characterized by size, zeta potential (ZP), polydispersity index (Pdi), cytotoxicity and mucoadhesion. Encapsulation efficiency and drug loading capacity were also determined. Ex vivo permeation was tested using fresh porcine corneas, scleras and conjunctivas. The permeated EPO was quantified by ELISA, and its presence in the membranes was confirmed by immunohistochemistry. Nanoparticles (NPs) presented size ≤300 nm, ZP around +30 mV and low Pdi (0.167-0.539) at a 1:1 CS:HA mass ratio. The most suitable HA was HA6 (300 kDa - Eye), which had the best mucoadhesive properties. CS/HA6-EPO nanoformulation permeated more rapidly through porcine conjunctiva, followed by sclera and thirdly by cornea, as assessed by immunohistochemistry. All formulations were noncytotoxic on ARPE-19 and HaCaT cell lines, as evaluated by metabolic and membrane integrity tests. In conclusion, CS/HA6-EPO NPs could be a promising formulation for increasing EPO ocular bioavailability by enhancing its retention time and permeation through the different ocular membranes.

Inhalable locust bean gum microparticles co-associating isoniazid and rifabutin: Therapeutic assessment in a murine model of tuberculosis infection.

Tuberculosis is a leading cause of death worldwide. Although the development of new antimycobacterial drugs is an obvious and necessary strategy to address the disease, improving the therapeutic performance of drugs already approved constitutes a valuable alternative approach. As the lung is the most affected organ, where M. tuberculosis is able to survive and proliferate, the direct pulmonary delivery of antitubercular drugs comprises a highly promising therapeutic strategy. In this work, spray-dried locust bean gum (LBG) microparticles were used to deliver a combination of two first line antitubercular drugs, isoniazid (INH) and rifabutin (RFB), to the alveolar zone, where macrophages hosting the bacteria reside. LBG is expected to mediate favoured macrophage uptake of microparticles, leading to enhanced therapeutic effect. The therapeutic effect of LBG/INH/RFB microparticles was evaluated in a murine model infected with M. tuberculosis, strain H37Rv and compared with oral co-therapy of INH and RFB in the free form. The pulmonary administration of LBG/INH/RFB microparticles 5 times per week was the only treatment schedule that provided negative growth index values in lung (-0.22), spleen (-0.14) and liver (-0.26) even using a lower therapeutic dose for both antibiotics. For the control group, the respective values were +1.95, +0.75 and +0.96.

Starch-Based Pickering Emulsions as Platforms for Topical Antibiotic Delivery: In Vitro and In Vivo Studies.

The present study investigated a new approach to treat superficial skin infections by topical application of minocycline hydrochloride (MH) formulated in a novel starch-based Pickering emulsion (ASt-emulsions). The emulsions were fully characterized in terms of efficacy, as well as in vitro release and permeation studies. The emulsions provided a prolonged MH release, always above its minimum inhibitory concentration against Staphylococcus aureus, although the drug did not permeate through the entire skin layer. The in vitro antibacterial activity of MHASt-emulsions against S. aureus was confirmed and their therapeutic efficacy was assessed using an in vitro skin-adapted agar diffusion test. In vivo antibacterial activity, evaluated using the tape-stripping infection model in mice, showed the topical administration of MH was effective against superficial infections caused by S. aureus. This study supports the potential of ASt-emulsions as promising platforms for topical antibiotic delivery, contributing to a new perspective on the treatment of superficial bacterial infections.

Combination of hyaluronic acid and PLGA particles as hybrid systems for viscosupplementation in osteoarthritis.

Hyaluronic acid (HA) is commonly used through intra-articular administration for viscosupplementation in osteoarthritis and other disorders. HA is generally supplied as an injection commonly reported as painful, with strong limitations after treatment. In this study, an alternative delivery system was constructed based on HA hydrogel and poly(lactic-co-glycolic acid) (PLGA) particles with oleic acid. Development studies included the determination of particle toxicity, hemolytic activity, in vitro and in vivo anti-inflammatory activity using macrophages and a murine model, respectively. This study showed that empty PLGA particles presented a mean size of 373 nm, while particles containing HA and oleic acid showed a marked particle size increase. The HA association efficiency was of 73.6% and 86.2% for PLGA particles without and with oleic acid, respectively. The in vitro HA release from PLGA particles revealed a sustained profile. Particles showed a good in vitro cell compatibility and the risk of hemolysis was less <1%, ensuring their safety. The in vivo anti-inflammatory study showed a higher inhibition for HA-loaded PLGA particles when compared to HA solution (78% versus 60%) and they were not different from the positive control, clearly suggesting that this formulation may be a promising alternative to the current HA commercial dosage form.

Associative organization of nursing: struggles for the social recognition of the profession (1943-1946).

OBJECTIVE:: To describe the circumstances that determine the creation of the Brazilian Association of Graduate Nurses of the Federal District Section and analyze its implications for the reorganization of the field of nursing. METHOD:: Qualitative, socio-historical, documentary study. The analysis generated the following category: Creation of a new group to guarantee unity: Brazilian Association of Graduate Nurses in the Federal District Section. RESULTS:: The economic crisis resulting from the Second World War, the creation of the Paulista Association of Graduate Nurses and the increase in the number of Schools of Nursing in the country were decisive for the Brazilian Association of Graduate Nurses to reformulate its statute as to guarantee its unit. FINAL CONSIDERATIONS:: The creation of the Federal District Section consisted of one of the strategies of the Association to reorganize the field of nursing, in order to ensure the recognition of the profession by the society. OBJETIVOS:: Descrever as circunstâncias que determinam a criação da Associação Brasileira de Enfermeiras Diplomadas Seção do Distrito Federal e analisar as implicações dessa criação para a reorganização do campo da enfermagem. MÉTODO:: Estudo qualitativo, sócio-histórico, documental. A análise gerou a seguinte categoria: Criação de um novo grupo para garantir a unidade: Associação Brasileira de Enfermeiras Diplomadas Seção do Distrito Federal. RESULTADOS:: A crise econômica, decorrente da Segunda Guerra Mundial, a criação da Associação Paulista de Enfermeiras Diplomadas e o aumento do número de escolas de enfermagem no país foram determinantes para que a Associação Brasileira de Enfermeiras Diplomadas reformulasse seu estatuto, de modo a garantir sua unidade. CONSIDERAÇÕES FINAIS:: A criação da Seção do Distrito Federal consistiu em uma das estratégias da associação para reorganizar o campo da enfermagem, de modo a assegurar o reconhecimento da profissão pela sociedade.

Approaches to tuberculosis mucosal vaccine development using nanoparticles and microparticles: a review.

Next-generation vaccines for tuberculosis should be designed to prevent the infection and to achieve sterile eradication of Mycobacterium tuberculosis. Mucosal vaccination is a needle-free vaccine strategy that provides protective immunity against pathogenic bacteria and viruses in both mucosal and systemic compartments, being a promising alternative to current tuberculosis vaccines. Micro and nanoparticles have shown great potential as delivery systems for mucosal vaccines. In this review, the immunological principles underlying mucosal vaccine development will be discussed, and the application of mucosal adjuvants and delivery systems to the enhancement of protective immune responses at mucosal surfaces will be reviewed, in particular those envisioned for oral and nasal routes of administration. An overview of the essential vaccine candidates for tuberculosis in clinical trials will be provided, with special emphasis on the potential different antigens and immunization regimens.

Advanced systems for glucocorticoids' dermal delivery.

INTRODUCTION: Although the skin provides a natural physical barrier against particle penetration, there are opportunities to deliver therapeutic agents through this barrier. With rapid developments in materials science, pharmaceutics, and biotechnology, new systems have emerged for topical glucocorticoids (TG) delivery. Despite being a mature class of drugs, TG are still the most frequently prescribed drugs by dermatologists, explaining the interest on this field. AREAS COVERED: Over the years, research has focused on strategies to optimize the potency of steroids while minimizing adverse effects. Several attempts have been made to increase the safety of TG treatment, including new application schedules, special vehicles, and new synthesized agents. This paper gives an overview on the conventional and advanced vehicles used to deliver TG, including strategies to improve the delivery and accumulation of TG. The therapeutic advantages of TG delivery to the epidermis are discussed with a special focus on new drug-delivery systems for topical application. EXPERT OPINION: New technologies have been developed for TG delivery, and studies are currently underway in order to obtain improved benefit/risk ratio. A rationale development approach that integrates simple formulations would help to develop vehicles with these features.

Polyol additives modulate the in vitro stability and activity of recombinant human phenylalanine hydroxylase.

Phenylketonuria (PKU; OMIM 261600), the most common disorder of amino acid metabolism, is caused by a deficient activity of human phenylalanine hydroxylase (hPAH). Although the dietetic treatment has proven to be effective in preventing the psycho-motor impairment, much effort has been made to develop new therapeutic approaches. Enzyme replacement therapy with hPAH could be regarded as a potential form of PKU treatment if the reported in vitro hPAH instability could be overcome. In this study, we investigated the effect of different polyol compounds, e.g. glycerol, mannitol and PEG-6000 on the in vitro stability of purified hPAH produced in a heterologous prokaryotic expression system. The recombinant human enzyme was stored in the presence of the studied stabilizing agents at different temperatures (4 and -20 degrees C) during a 1-month period. Protein content, degradation products, specific activity, oligomeric profile and conformational characteristics were assessed during storage. The obtained results showed that the use of 50% glycerol or 10% mannitol, at -20 degrees C, protected the enzyme from loss of its enzymatic activity. The determined DeltaG(0) and quenching parameters indicate the occurrence of conformational changes, which may be responsible for the observed increase in catalytic efficiency.

The quality of life of aged people living in homes for the aged / La calidad de vida de la persona de edad avanzada institucionalizada en hogares de ancianos / A qualidade de vida da pessoa idosa institucionalizada em lares

This is a descriptive cross-sectional study. The sample consisted of 93 aged people, divided among four institutions in the municipality of Lamego (Portugal). The purpose of this study was to investigate the quality of life of the sample and the factors affecting it, and identify the degree of dependence in basic daily living activities. The data was collected using the following: i) a bio-relational questionnaire, ii) a quality of life evaluation scale, and iii) the Katz Index. In terms of the quality of life of the institutionalized aged people, 51.6 percent include themselves in the group with quality of life, and it was observed there was a positive correlation between the Katz index and the quality of life index in the institutionalized aged people. There were no differences in the quality of life index regarding gender, marital status, degree of education, and the presence of pain in the institutionalized aged people.

Se trata de un estudio del tipo descriptivo y transversal, englobando una muestra con 93 ancianos, divididos en cuatro instituciones del Concejo de Lamego (Portugal), con el objetivo de conocer la calidad de vida, y los factores que la influencian, e identificar el grado de dependencia en las actividades básicas de la vida diaria. En la recolección de los datos se utilizó: i) formulario bio-relacional del anciano, ii) la planilla de evaluación de la calidad de vida del anciano, y iii) el índice de Katz. De las personas ancianas institucionalizadas, 51,6 por ciento están incluidas en el grupo con calidad de vida, existiendo una correlación positiva entre el índice de Katz y el índice de calidad de vida del anciano institucionalizado. El índice de calidad de vida no cambia en función del sexo, del estado civil, del nivel de instrucción y de la existencia de dolor de la persona anciana institucionalizada.

Trata-se de estudo do tipo descritivo e transversal, englobando amostra com 93 idosos, divididos por quatro instituições do Concelho de Lamego (Portugal), com o objetivo de conhecer a qualidade de vida, e fatores que a influenciam, e identificar o grau de dependência nas atividades básicas da vida diária. Na recolha de dados utilizou-se: i) formulário bio-relacional do idoso, ii) a grelha de avaliação da qualidade de vida do idoso e iii) o índice de Katz. Das pessoas idosas institucionalizadas, 51,6 por cento referem se incluir no grupo com qualidade de vida, existindo correlação positiva entre o índice de Katz e o índice de qualidade de vida do idoso institucionalizado. O índice de qualidade de vida não difere em função do sexo, estado civil, nível de instrução e da existência de dor da pessoa idosa institucionalizada.

The quality of life of aged people living in homes for the aged.

This is a descriptive cross-sectional study. The sample consisted of 93 aged people, divided among four institutions in the municipality of Lamego (Portugal). The purpose of this study was to investigate the quality of life of the sample and the factors affecting it, and identify the degree of dependence in basic daily living activities. The data was collected using the following: i) a bio-relational questionnaire, ii) a quality of life evaluation scale, and iii) the Katz Index. In terms of the quality of life of the institutionalized aged people, 51.6% include themselves in the group with quality of life, and it was observed there was a positive correlation between the Katz index and the quality of life index in the institutionalized aged people. There were no differences in the quality of life index regarding gender, marital status, degree of education, and the presence of pain in the institutionalized aged people.