Evaluation of Aerosol Therapy during the Escalation of Care in a Model of Adult Cystic Fibrosis.

Lung disease is the main cause of morbidity and mortality in cystic fibrosis (CF). CF patients inhale antibiotics regularly as treatment against persistent bacterial infections. The goal of this study was to investigate the effect of clinical intervention on aerosol therapy during the escalation of care using a bench model of adult CF. Droplet size analysis of selected antibiotics was completed in tandem with the delivered aerosol dose (% of total dose) assessments in simulations of various interventions providing oxygen supplementation or ventilatory support. Results highlight the variability of aerosolised dose delivery. In the homecare setting, the vibrating mesh nebuliser (VMN) delivered significantly more than the jet nebuliser (JN) (16.15 ± 0.86% versus 6.51 ± 2.15%). In the hospital setting, using VMN only, significant variability was seen across clinical interventions. In the emergency department, VMN plus mouthpiece (no supplemental oxygen) was seen to deliver (29.02 ± 1.41%) versus low flow nasal therapy (10 L per minute (LPM) oxygen) (1.81 ± 0.47%) and high flow nasal therapy (50 LPM oxygen) (3.36 ± 0.34%). In the ward/intensive care unit, non-invasive ventilation recorded 19.02 ± 0.28%, versus 22.64 ± 1.88% of the dose delivered during invasive mechanical ventilation. These results will have application in the design of intervention-appropriate aerosol therapy strategies and will be of use to researchers developing new therapeutics for application in cystic fibrosis and beyond.

Precise Targeting of miRNA Sites Restores CFTR Activity in CF Bronchial Epithelial Cells.

MicroRNAs that are overexpressed in cystic fibrosis (CF) bronchial epithelial cells (BEC) negatively regulate CFTR and nullify the beneficial effects of CFTR modulators. We hypothesized that it is possible to reverse microRNA-mediated inhibition of CFTR using CFTR-specific target site blockers (TSBs) and to develop a drug-device combination inhalation therapy for CF. Lead microRNA expression was quantified in a series of human CF and non-CF samples and in vitro models. A panel of CFTR 3' untranslated region (UTR)-specific locked nucleic acid antisense oligonucleotide TSBs was assessed for their ability to increase CFTR expression. Their effects on CFTR activity alone or in combination with CFTR modulators were measured in CF BEC models. TSB encapsulation in poly-lactic-co-glycolic acid (PLGA) nanoparticles was assessed as a proof of principle of delivery into CF BECs. TSBs targeting the CFTR 3' UTR 298-305:miR-145-5p or 166-173:miR-223-3p sites increased CFTR expression and anion channel activity and enhanced the effects of ivacaftor/lumacaftor or ivacaftor/tezacaftor in CF BECs. Biocompatible PLGA-TSB nanoparticles promoted CFTR expression in primary BECs and retained desirable biophysical characteristics following nebulization. Alone or in combination with CFTR modulators, aerosolized CFTR-targeting TSBs encapsulated in PLGA nanoparticles could represent a promising drug-device combination therapy for the treatment for CFTR dysfunction in the lung.

CFTR dysfunction in cystic fibrosis and chronic obstructive pulmonary disease.

INTRODUCTION: Obstructive lung diseases such as cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD) are causes of high morbidity and mortality worldwide. CF is a multiorgan genetic disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene and is characterized by progressive chronic obstructive lung disease. Most cases of COPD are a result of noxious particles, mainly cigarette smoke but also other environmental pollutants. Areas covered: Although the pathogenesis and pathophysiology of CF and COPD differ, they do share key phenotypic features and because of these similarities there is great interest in exploring common mechanisms and/or factors affected by CFTR mutations and environmental insults involved in COPD. Various molecular, cellular and clinical studies have confirmed that CFTR protein dysfunction is common in both the CF and COPD airways. This review provides an update of our understanding of the role of dysfunctional CFTR in both respiratory diseases. Expert commentary: Drugs developed for people with CF to improve mutant CFTR function and enhance CFTR ion channel activity might also be beneficial in patients with COPD. A move toward personalized therapy using, for example, microRNA modulators in conjunction with CFTR potentiators or correctors, could enhance treatment of both diseases.

Biopolymer-Based Nanoparticles for Cystic Fibrosis Lung Gene Therapy Studies.

Lung gene therapy for cystic fibrosis disease has not been successful due to several challenges such as the absence of an appropriate vector. Therefore, optimal delivery of emerging therapeutics to airway epithelial cells demands suitable non-viral systems. In this work, we describe the formulation and the physicochemical investigation of biocompatible and biodegradable polymeric nanoparticles (NPs), including PLGA and chitosan (animal and non-animal), as novel methods for the safe and efficient delivery of CFTR-specific locked nucleic acids (LNAs).

Chitosan in Non-Viral Gene Delivery: Role of Structure, Characterization Methods, and Insights in Cancer and Rare Diseases Therapies.

Non-viral gene delivery vectors have lagged far behind viral ones in the current pipeline of clinical trials of gene therapy nanomedicines. Even when non-viral nanovectors pose less safety risks than do viruses, their efficacy is much lower. Since the early studies to deliver pDNA, chitosan has been regarded as a highly attractive biopolymer to deliver nucleic acids intracellularly and induce a transgenic response resulting in either upregulation of protein expression (for pDNA, mRNA) or its downregulation (for siRNA or microRNA). This is explained as the consequence of a multi-step process involving condensation of nucleic acids, protection against degradation, stabilization in physiological conditions, cellular internalization, release from the endolysosome ("proton sponge" effect), unpacking and enabling the trafficking of pDNA to the nucleus or the siRNA to the RNA interference silencing complex (RISC). Given the multiple steps and complexity involved in the gene transfection process, there is a dearth of understanding of the role of chitosan's structural features (Mw and degree of acetylation, DA%) on each step that dictates the net transfection efficiency and its kinetics. The use of fully characterized chitosan samples along with the utilization of complementary biophysical and biological techniques is key to bridging this gap of knowledge and identifying the optimal chitosans for delivering a specific gene. Other aspects such as cell type and administration route are also at play. At the same time, the role of chitosan structural features on the morphology, size and surface composition of synthetic virus-like particles has barely been addressed. The ongoing revolution brought about by the recent discovery of CRISPR-Cas9 technology will undoubtedly be a game changer in this field in the short term. In the field of rare diseases, gene therapy is perhaps where the greatest potential lies and we anticipate that chitosans will be key players in the translation of research to the clinic.

Chitosan as a non-viral co-transfection system in a cystic fibrosis cell line.

Successful gene therapy requires the development of suitable vehicles for the selective and efficient delivery of genes to specific target cells at the expense of minimal toxicity. In this work, we investigated a non-viral gene delivery system based on chitosan (CS) to specifically address cystic fibrosis (CF). Thus, electrostatic self-assembled CS-pEGFP and CS-pEGFP-siRNA complexes were prepared from high-pure fully characterized CS (Mw ∼ 20 kDa and degree of acetylation ∼ 30%). The average diameter of positively-charged complexes (i.e. ζ ∼+25 mV) was ∼ 200 nm. The complexes were found relatively stable over 14h in Opti-MEM. Cell viability study did not show any significant cytotoxic effect of the CS-based complexes in a human bronchial cystic fibrosis cell line (CFBE41o-). We evaluated the transfection efficiency of this cell line with both CS-pEGFP and co-transfected with CS-pEGFP-siRNA complexes at (N/P) charge ratio of 12. We reported an increase in the fluorescence intensity of CS-pEGFP and a reduction in the cells co-transfected with CS-pEGFP-siRNA. This study shows proof-of-principle that co-transfection with chitosan might be an effective delivery system in a human CF cell line. It also offers a potential alternative to further develop therapeutic strategies for inherited disease treatments, such as CF.

Documento de Consenso: importancia nutricional y metabólica de la leche / Consensus document: nutritional and metabolic importance of cow´s milk

La leche de vaca es un alimento básico en la alimentación humana en todas las etapas de la vida. Su procesamiento industrial ha permitido el acceso generalizado a su consumo por parte de la población, lo que ha contribuido a mejorar notablemente su nivel de salud. Desde el punto de vista de su composición, la leche es un alimento completo y equilibrado, que proporciona un elevado contenido de nutrientes en relación con su contenido calórico, por lo que su consumo debe considerarse necesario desde la infancia a la tercera edad. Los beneficios de la leche de vaca no se limitan exclusivamente a su valor nutricional, sino que se extienden más allá y constituyen un factor de prevención en determinadas patologías afluentes como son la enfermedad cardiovascular, algunos tipos de cáncer, la hipertensión arterial o en patología ósea o dental. Puede contribuir también en la lucha frente al sobrepeso y la obesidad infantil. En los últimos años hemos asistido a un descenso preocupante en el consumo de leche entre la población española, condicionado al menos en parte por ideas equivocadas sobre su consumo y el de otros derivados lácteos. Este documento de consenso pretende revisar el estado actual de la cuestión en relación con los efectos del consumo de leche sobre la salud, al tiempo que hace una llamada a las instituciones y a las sociedades científicas para elaborar programas y campañas divulgativas sobre los beneficios del consumo de leche y derivados lácteos (AU)

Cow’s milk is a staple food for human consumption at all stages of life. Industrial processing has allowed widespread access to its consumption by the population, which has helped to significantly improve their health. From its composition point of view, milk is a complete and balanced food that provides high nutrient content in relation to its calorie content, so its consumption should be considered necessary from childhood to elderly. The benefits of cow’s milk are not limited to its nutritional value, but extend beyond and are a factor of prevention in certain non communicable pathologies as cardiovascular disease, some cancers, high blood pressure or bone or dental pathology. It can also help in the fight against childhood overweight and obesity. In recent years we have seen a worrying decline in milk consumption among the Spanish population, at least in part influenced by misconceptions about its consumption and of other dairy products. This consensus document aims to review the current state of the topic regarding the effects of milk consumption on health, while making a call to the institutions and scientific societies to develop programs and information campaigns about the benefits of milk and dairy products consumption (AU)

[Consensus document: nutritional and metabolic importance of cow's milk]. / Documento de consenso: importancia nutricional y metabólica de la leche.

Cow's milk is a staple food for human consumption at all stages of life. Industrial processing has allowed widespread access to its consumption by the population, which has helped to significantly improve their health. From its composition point of view, milk is a complete and balanced food that provides high nutrient content in relation to its calorie content, so its consumption should be considered necessary from childhood to elderly. The benefits of cow's milk are not limited to its nutritional value, but extend beyond and are a factor of prevention in certain non communicable pathologies as cardiovascular disease, some cancers, high blood pressure or bone or dental pathology. It can also help in the fight against childhood overweight and obesity. In recent years we have seen a worrying decline in milk consumption among the Spanish population, at least in part influenced by misconceptions about its consumption and of other dairy products. This consensus document aims to review the current state of the topic regarding the effects of milk consumption on health, while making a call to the institutions and scientific societies to develop programs and information campaigns about the benefits of milk and dairy products consumption.

La leche de vaca es un alimento básico en la alimentación humana en todas las etapas de la vida. Su procesamiento industrial ha permitido el acceso generalizado a su consumo por parte de la población, lo que ha contribuido a mejorar notablemente su nivel de salud. Desde el punto de vista de su composición, la leche es un alimento completo y equilibrado, que proporciona un elevado contenido de nutrientes en relación con su contenido calórico, por lo que su consumo debe considerarse necesario desde la infancia a la tercera edad. Los beneficios de la leche de vaca no se limitan exclusivamente a su valor nutricional, sino que se extienden más allá y constituyen un factor de prevención en determinadas patologías afluentes como son la enfermedad cardiovascular, algunos tipos de cáncer, la hipertensión arterial o en patología ósea o dental. Puede contribuir también en la lucha frente al sobrepeso y la obesidad infantil. En los últimos años hemos asistido a un descenso preocupante en el consumo de leche entre la población española, condicionado al menos en parte por ideas equivocadas sobre su consumo y el de otros derivados lácteos. Este documento de consenso pretende revisar el estado actual de la cuestión en relación con los efectos del consumo de leche sobre la salud, al tiempo que hace una llamada a las instituciones y a las sociedades científicas para elaborar programas y campañas divulgativas sobre los beneficios del consumo de leche y derivados lácteos.

Cystic fibrosis transmembrane conductance regulator-mRNA delivery: a novel alternative for cystic fibrosis gene therapy.

BACKGROUND: Cystic fibrosis (CF) is the most frequent lethal genetic disease in the Caucasian population. CF is caused by a defective gene coding for the cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP- and ATP-dependent Cl(-) channel and central regulatory protein in epithelia. CFTR influences the fluid composition of the mucus in the respiratory tract. The most common mutation inducing CF, ΔF508, impairs CFTR processing within the cell and thus prevents functional CFTR expression in the apical membrane. The present study aimed to investigate the functional restoration of CFTR in human CF airway epithelia after transfection with optimized wild-type (wt)CFTR-mRNA. METHODS: We used primary cultured human nasal epithelial (HNE) cells and the human bronchial epithelial cell line CFBE41o(-) that stably expresses ΔF508-CFTR and carried out transepithelial Ussing chamber measurements after transfection with optimized wtCFTR-mRNA. We confirmed the data obtained using immunofluorescence and protein biochemical approaches. RESULTS: Transfection of the CFBE41o(-) cells with wtCFTR-mRNA restored cAMP-induced CFTR currents similar to the values seen in control cells (16HBE14o(-)). Using immunofluorescence approaches, we demonstrated that a considerable amount of CFTR is located at the apical surface in the CF cells after transfection. Western blot analyses of wtCFTR-mRNA transfected CFBE41o(-) cells confirmed these findings. Furthermore, we demonstrated physiological relevance by using primary cultured HNE cells and showed an almost two-fold increase in the cAMP-stimulated CFTR current after transfection. CONCLUSIONS: From these data, we conclude that CFTR-mRNA transfection could comprise a novel alternative for gene therapy to restore impaired CFTR function.

Calidad de vida después recidiva bioquímica en los pacientes con cáncer de próstata. ¿Cómo y cuánto viven los pacientes después de recidiva bioquímica? / Quality of life after biochemical recurrence in patients with prostate cancer. How and how long do patients live after biochemical recurrence?

La monitorización rutinaria de PSA en pacientes con cáncer de próstata localizado tratados radicalmente permite identificar aquellos con recurrencia bioquímica exclusiva. Las opciones de tratamiento para el fallo bioquímico incluyen observación, cirugía, radioterapia externa sola o asociada a hormonoterapia, braquiterapia, crioterapia y hormonoterapia exclusiva. Estos tratamientos determinan un patrón específico de cambios (función urinaria, intestinal, sexual y hormonal) que puede impactar negativamente sobre la calidad de vida, de manera que su indicación debe realizarse de una manera juiciosa y siempre en consonancia con las expectativas y preferencias de los pacientes. Las decisiones sobre cómo y cuándo tratar un fallo bioquímico son complicadas y el impacto de las terapias de rescate sobre el resultado clínico final es desconocido. Las tasas de control del cáncer de próstata tras tratamiento de rescate con prostatectomía, crioterapia ó braquiterapia oscilan entre el 20-80% de los casos en función de las características de los pacientes seleccionados. Dado que los individuos con fallo bioquímico pueden estar clínicamente asintomáticos durante muchos años sin tratamiento es esencial que médicos y pacientes dispongan de un claro conocimiento del potencial impacto de las mismas sobre la calidad de vida(AU)

Routine monitoring of PSA in patients with localized prostate cancer radically treated permits to identify those with biochemical recurrence only. Treatment options for biochemical failure include observation, surgery, radiotherapy alone or combined with hormonal therapy, brachytherapy, cryotherapy and hormone therapy exclusively. These treatments determine a specific pattern of changes (urinary function, bowel, sexual and hormonal) that can negatively impact the quality of life, so that the indication must be made in a judicious way and always in consonance with patient’s expectations and preferences. Decisions on how and when to treat biochemical failure are complicated and the impact of salvage therapy on clinical outcome is unknown. Rates of prostate cancer control after salvage therapy with prostatectomy, brachytherapy or cryotherapy vary between 20-80% of cases according to selected patient characteristics. Because individuals with BF may be clinically asymptomatic for many years without treatment, it is essential that physicians and patients have a clear understanding of the potential impact of these on the quality of life(AU)