In Vitro Characterization and Real-Time Label-Free Assessment of the Interaction of Chitosan-Coated Niosomes with Intestinal Cellular Monolayers.

In vitro cell-based characterization methods of nanoparticles are generally static and require the use of secondary analysis techniques and labeling agents. In this study, bare niosomes and chitosan-coated niosomes (chitosomes) and their interactions with intestinal cells are studied under dynamic conditions and without fluorescent probes, using surface plasmon resonance (SPR)-based cell sensing. Niosomes and chitosomes were synthesized by using Tween 20 and cholesterol in a 15 mM:15 mM ratio and then characterized by dynamic light scattering (DLS). DLS analysis demonstrated that bare niosomes had average sizes of ∼125 nm, polydispersity index (PDI) below 0.2, and a negative zeta (ζ)-potential of -35.6 mV. In turn, chitosomes had increased sizes up to ∼180 nm, with a PDI of 0.2-0.3 and a highly positive ζ-potential of +57.9 mV. The viability of HT29-MTX, Caco-2, and Caco-2/HT29-MTX cocultured cells showed that both niosomes and chitosomes are cytocompatible up to concentrations of 31.6 µg/mL for at least 240 min. SPR analysis demonstrated that chitosomes interact more efficiently with HT29-MTX, Caco-2, and Caco-2/HT29-MTX cocultures compared to bare niosomes. The resulting SPR measurements were further supported by confocal microscopy and flow cytometry studies, which demonstrated that this method is a useful complementary or even alternative tool to directly characterize the interactions between niosomes and in vitro cell models in label-free and real-time conditions.

Microfluidic-Assisted Production of Gastro-Resistant Active-Targeted Diatomite Nanoparticles for the Local Release of Galunisertib in Metastatic Colorectal Cancer Cells.

The oral route is highly desirable for colorectal cancer (CRC) treatment because it allows concentrating the drug in the colon and achieving a localized effect. However, orally administered drugs are often metabolized in the liver, resulting in reduced efficacy and the need for higher doses. Nanoparticle-based drug delivery systems can be engineered to prevent the diffusion of the drug in the stomach, addressing the release at the target site, and enhancing the efficacy of the delivered drug. Here, an orally administrable galunisertib delivery system is developed with gelatin-covered diatomite nanoparticles targeting the ligand 1-cell adhesion molecule (L1-CAM) on metastatic cells, and further encapsulated in an enteric matrix by microfluidics. The gastro-resistant polymer protects the nanoparticles from the action of the digestive enzymes and allows for a sustained release of galunisertib at the intestinal pH. The efficacy of antibody-antigen interactions to drive the internalization of nanoparticles in the targeted cells is investigated in CRC cells expressing abnormal (SW620) or basal levels (Caco-2, HT29-MTX) of L1-CAM. The combination of local drug release and active targeting enhances the effect of the delivered galunisertib, which inhibits the migration of the SW620 cells with greater efficiency compared to the free drug.

Analysis of pain relief and functional recovery in patients with rotator cuff tendinopathy through therapeutic ultrasound and photobiomodulation therapy: a comparative study.

This study aimed to compare shoulder tendinopathy treatment with therapeutic ultrasound combined with LED photobiomodulation therapy using LED-infrared (850 nm) or LED-red (640 nm). The study assessed 75 patients, aged 45 to 70 years, distributed into five experimental groups (15 patients each): therapeutic ultrasound (US), infrared light irradiation (IR), visible red light irradiation (VR), infrared light and ultrasound combined (IR-US), and red light in conjunction with ultrasound (VR-US). The ultrasound parameters are 1 MHz, 0.5 W/cm2 (SATA), and 100 Hz repetition rate, applied for 4 min each session. LED irradiation protocols were as follows: 3 points, 7.5 J per point, IR-LED 750 mW, 10 s, VR-LED 250 mW, 30 s. LED irradiation is followed by ultrasound in the combined therapies. The efficiency of the five therapies was evaluated assessing 12 parameters: quality of life (Health Assessment Questionnaire, HAQ), pain intensity (Visual Analog Scale, VAS), articular amplitude of shoulder movement (flexion, extension, abduction, adduction, medial rotation, lateral rotation), muscle strength (abduction, lateral rotation), and electromyography (lateral rotation, abduction). Treatments comprised 12 sessions for 4 weeks. Intra-group analysis showed that the five therapies significantly improved the recovery of all parameters after treatment. Regarding the comparison of irradiated therapies and ultrasound, statistical analysis showed that IR-US was a better treatment than US for all 12 parameters. IR treatment exceeded US on 9 items, whereas that VR and VR-US therapies exceeded US in 7 and 10 parameters, respectively (p < 0.05). Because of that, IR-US shows to be the best treatment for rotator cuff tendinopathy. In conclusion, improvements in quality of life, pain intensity relief, shoulder amplitude motion, and muscle strength force obtained with ultrasound therapy are enhanced by adding infrared LED irradiation to ultrasound for patients suffering from rotator cuff tendinopathy. This study was registered with the Brazilian Registry of Clinical Trials (ReBEC) under Universal Trial Number (UTN) U1111-1219-3594 (2018/22/08).

[Early Detection of COVID-19 in Portugal: Use of Clinical Records]. / Deteção Precoce de COVID-19 em Portugal: Uso de Registos Clínicos.

INTRODUCTION: Syndromic surveillance allows early detection of changes in the population's morbidity pattern. The aim of this study is to evaluate the usefulness of indicators related to access to healthcare services, in COVID-19 surveillance. MATERIAL AND METHODS: A time series analysis was performed using the weekly incidence rate of COVID-19 in Mainland Portugal, between weeks 14/2020 (March 30 to April 5) and 25/2020 (June 15 to 21), and six indicators: 1) COVID-19 consultations in primary healthcare; 2) number of COVID-19 emergency department visits; 3) number of emergency department visits due to viral pneumonia; 4) number of hospitalizations due to viral pneumonia; 5) proportion of emergency department visits due to viral pneumonia; and 6) proportion of hospitalizations for viral pneumonia. Pearson correlation and cross-correlations were computed. RESULTS: A strong correlation was found between the weekly incidence rate of COVID-19 and all indicators. [(1) 0.76; (2) 0.82; (3) 0.77; (4) 0.84; (5) 0.86; e (6) 0.90]. Emergency department visits and hospitalizations for viral pneumonia detect variations in the frequency of the disease with a one week lag compared to the incidence rate of COVID-19, in one week. COVID-19 consultations in primary healthcare and emergency department visits trail behind the incidence rate of COVID-19, in one week. The proportion of viral pneumonias in emergency department visits, or hospitalizations, is temporally aligned with the weekly incidence rate of COVID-19. DISCUSSION: The delay found in the COVID-19 primary healthcare consultations and emergency department visits, may be related to changes in access to healthcare services and clinical coding. Emergency department visits and hospitalizations for viral pneumonia may be useful in the early detection of COVID-19. Viral pneumonia may have been coded as being of unknown origin. Future monitoring of these indicators is necessary to ascertain whether the incidence of COVID-19 is significantly influenced by changes in testing strategies. The indicators described in this study will be an asset for the optimization of testing strategies, allocation of healthcare resources to the communities that are most vulnerable to severe morbidity and assessing vaccination impact. As such, surveillance systems based on clinical data will be a valuable complementary tool to SINAVE. CONCLUSION: The indicators under analysis could be used regularly, with special attention to viral pneumonias, to detect outbreaks of COVID-19. Information on pneumonia of unknown etiology may be considered in the surveillance of COVID-19.

Introdução: A vigilância sindrómica permite a identificação precoce de alterações no padrão de morbilidade da população. Este estudo tem como objetivo avaliar a utilidade de indicadores relativos a cuidados de saúde primários e hospitalares, na vigilância da COVID-19.Material e Métodos: Foi realizada uma análise de séries temporais utilizando a taxa de incidência semanal de COVID-19 em Portugal Continental, entre as semanas 14/2020 (30 março a 05 abril) e 25/2020 (15 a 21 junho), e seis indicadores: 1) consultas em cuidados de saúde primários por COVID-19; 2) número de episódios de urgência por COVID-19; 3) número de episódios de urgência por pneumonia vírica; 4) número de internamentos por pneumonia vírica; 5) proporção de episódios de urgência por pneumonia vírica face ao total de episódios de urgência por pneumonia; e 6) proporção de internamentos por pneumonia vírica face ao total de internamentos por pneumonia. Foram calculadas correlações de Pearson e correlações cruzadas.Resultados: Foi encontrada uma correlação forte entre a taxa de incidência semanal de COVID-19 e todos os indicadores [(1) 0,76; (2) 0,82; (3) 0,77; (4) 0,84; (5) 0,86; e (6) 0,90]. Os episódios de urgência e internamento por pneumonias víricas detetam variações na frequência da doença, com uma semana de antecedência. As consultas em cuidados de saúde primários e urgências por COVID-19 registam uma semana de atraso relativamente à evolução da taxa de incidência. A proporção de pneumonias víricas face ao número de pneumonias em episódios de urgência, ou internamentos, encontra-se alinhada temporalmente com a evolução da taxa de incidência semanal de COVID-19.Discussão: O atraso encontrado no padrão de evolução de consultas em CSP, e de episódios de urgência por COVID-19 face à incidência de COVID-19, poderá estar relacionado com a reorganização dos serviços de saúde e criação de códigos específicos para estas consultas. Episódios de urgência e internamentos por pneumonia vírica poderão ser úteis para a deteção precoce de possíveis surtos de COVID-19. Pneumonias víricas poderão ter sido classificadas como pneumonias de causa indeterminada. A monitorização futura destes indicadores é necessária de modo a averiguar se a incidência de COVID-19 é influenciada significativamente por alterações na estratégia de testagem. Os indicadores deste trabalho serão uma mais valia para a adequação de estratégias de testagem, alocação de recursos de saúde a comunidades mais vulneráveis à morbilidade severa e avaliação de programas de vacinação. Como tal, os sistemas de vigilância com base em registos de saúde serão um complemento valioso ao SINAVE.Conclusão: Sugere-se que os indicadores em análise sejam utilizados de forma regular, com especial atenção à informação relativa a pneumonias víricas, como forma de detetar precocemente surtos de COVID-19. A informação relativa a pneumonias de causa indeterminada poderá ser considerada na monitorização da COVID-19.

The solid progress of nanomedicine.

This commentary article conveys the views of the board of the Nanomedicine and Nanoscale Delivery Focus Group of the Controlled Release Society regarding the decision of the United States National Cancer Institute (NCI) in halting funding for the Centers of Cancer Nanotechnology Excellence (CCNEs), and the subsequent editorial articles that broadened this discussion. Graphical abstract.

Avaliação da contaminação de nitrato e Escherichia coli em água subterrânea no município de Carapicuiba (SP) / Evaluation of nitrate and Escherichia coli contamination on groundwater in the city of Carapicuiba (SP)

Objetivo ­ Verificar a concentração de nitrato e a presença ou ausência de bactérias da espécie Escherichia coli em amostras de água subterrânea (poços) destinadas ao consumo humano e comparar os resultados aos Valores Máximos Permitidos (VMP) pela Portaria 2.914 do Ministério da Saúde. Métodos ­ Durante um ano (2017-2018), semestralmente, foram coletadas amostras de águasconsumidas por moradores em diferentes pontos: poços 1 e 2, ambos sem tratamento e com o auxílio de amostradores descartáveis do tipo bailer e; torneiras, denominados S1 e S2, água tratada pela Companhia de Saneamento Básico do Estado de São Paulo (SABESP) e destinadas ao abastecimento público. Esses últimos como grupo controle, pois não foram identificados poços de monitoramento ou nascentes próximas sem contaminação. As amostras em frascos apropriados foram preservadas em caixas térmicas (± 4°C) e encaminhadas para análise química e microbiológica, seguindo os protocolos de coletas da CETESB e USEPA e os métodos 4500-NO3-B e 9215c da APHA. As análises físico-químicas, tais como pH, OD, temperatura, cloro livre, condutividade elétrica e profundidade foram realizadas em campo com equipamentos, tais como multiparâmetro da marca Hanna Hi 9828 e outros. Resultados ­ Verificou-se que as águas subterrâneas são de natureza rasas (poços com profundidade < 20 m) e suas águas ligeiramente ácida (pH=5,0). Em ambos os poços houve a presença de bactérias E. coli e elevada concentração do íon nitrato (>18 mg/L) no período seco e chuvoso, estando em não conformidade ao VMP dado pela Portaria 2.914. As amostras S1 e S2 apresentaram ausência de E.coli e valores abaixo de 0,5 mg/L de N-NO-3. informação dos parâmetros de qualidade nos rótulos. O levantamento de alguns rótulos, mostrou que em cinco marcas de água envasadas, três delas estão não conforme a VMP ora citada (NO-3-N > 10 mg/L). Conclusão ­ O presente estudo demonstrou que para fins de potabilidade a água dos poços está imprópria para o consumo humano

Objective ­ To verify nitrate concentration, the presence or absence of Escherichia coli bacterium in groundwater samples (wells) intended for human consumption, and to compare the results to Maximum Permitted Values (MPV) by Portaria 2.914 of the Ministry of Health. Methods ­ During one year (2017-2018), samples of water consumed by residents at different points were taken every semester: wells 1 and 2, both without treatment and with the aid of disposable samplers of the bailer type; taps, called S1 and S2, water treated by the Basic Sanitation Company of the State of São Paulo (SABESP) and intended for public supply. The latter as a control group because no monitoring wells or nearby springs were identified without contamination. The samples in appropriate flasks were preserved in thermal boxes (± 4°C) and sent for chemical and microbiological analysis, following CETESB and USEPA collection protocols and APHA methods 4500-NO3-B and 9215c. Physical-chemical analyzes, such as pH, OD, temperature, free chlorine, electrical conductivity and depth were performed in the field with equipment such as multi-parameter, Hanna brand and others. Results ­ Groundwaters were found to be shallow (wells with depth <20 m) and their waters slightly acidic (pH = 5.0). In both wells, the presence of E. coli bacteria and high concentration of nitrate ion (> 18 mg / L) in the dry and rainy season were observed, being in non-compliance with the MPV given by Portaria 2.914. Samples S1 and S2 showed absence of E.coli and values below 0.5 mg / L of N-NO-3. Information on the quality parameters on the labels. The survey of some labels showed that in five bottled watermarks, three of them are contaminated by nitrate according to the MPV (NO-3-N> 10 mg / L). Conclusion ­ The present study demonstrates that drinking water from wells is unfit for human consumption.

The importance of microfluidics for the preparation of nanoparticles as advanced drug delivery systems.

INTRODUCTION: Nanoparticles are anticipated to overcome persistent challenges in efficient drug delivery, but the limitations associated with conventional methods of preparation are resulting in slow translation from research to clinical applications. Due to their enormous potential, microfluidic technologies have emerged as an advanced approach for the development of drug delivery systems with well-defined physicochemical characteristics and in a reproducible manner. AREAS COVERED: This review provides an overview of microfluidic devices and materials used for their manufacturing, together with the flow patterns and regimes commonly used for nanoparticle preparation. Additionally, the different geometries used in droplet microfluidics are reviewed, with particular attention to the co-flow geometry used for the production of nanoparticles. Finally, this review summarizes the main and most recent nanoparticulate systems prepared using microfluidics, including drug nanosuspensions, polymeric, lipid, structured, and theranostic nanoparticles. EXPERT OPINION: The production of nanoparticles at industrial scale is still a challenge, but the microfluidic technologies bring exciting opportunities to develop drug delivery systems that can be engineered in an easy, cost-effective and reproducible manner. As a highly interdisciplinary research field, more efforts and general acceptance are needed to allow for the translation of nanoparticulate drug delivery systems from academic research to the clinical practice.

Dynamics of binary mixtures of an ionic liquid and ethanol by NMR.

A study of molecular dynamics of the ionic liquid 1-ethyl-3-methylimidazolium bis(trifluoro-methylsulphonyl)imide ([Emim][Tf2N]) in solution with deuterated ethanol at different molar concentration and temperatures is presented. The study was performed using 1 H and 2 H nuclear magnetic relaxation and 2 H 1D spectroscopy. The temperature dependence of the spin-lattice relaxation time T1 of the cations allows the evaluation of the activation energies of the rotational degree of freedom of these molecules. The viscosity in the binary system increases with the concentration of ionic liquid. However, the activation energy in the cation molecules decreases when the concentration of the ionic liquid increases, indicating that the rotational dynamics is facilitated. This behavior is explained from the fact that the presence of the ionic liquid in the system disrupts the degree of intermediate range order expected in the alcohol system. Copyright © 2017 John Wiley & Sons, Ltd.

The Translation of Cyclin B1 and B2 is Differentially Regulated during Mouse Oocyte Reentry into the Meiotic Cell Cycle.

Control of protein turnover is critical for meiotic progression. Using RiboTag immunoprecipitation, RNA binding protein immunoprecipitation, and luciferase reporter assay, we investigated how rates of mRNA translation, protein synthesis and degradation contribute to the steady state level of Cyclin B1 and B2 in mouse oocytes. Ribosome loading onto Ccnb1 and Mos mRNAs increases during cell cycle reentry, well after germinal vesicle breakdown (GVBD). This is followed by the translation of reporters containing 3' untranslated region of Mos or Ccnb1 and the accumulation of Mos and Cyclin B1 proteins. Conversely, ribosome loading onto Ccnb2 mRNA and Cyclin B2 protein level undergo minimal changes during meiotic reentry. Degradation rates of Cyclin B1 or B2 protein at the GV stage are comparable. The translational activation of Mos and Ccnb1, but not Ccnb2, mRNAs is dependent on the RNA binding protein CPEB1. Inhibition of Cdk1 activity, but not Aurora A kinase activity, prevents the translation of Mos or Ccnb1 reporters, suggesting that MPF is required for their translation in mouse oocytes. Conversely, Ccnb2 translation is insensitive to Cdk1 inhibition. Thus, the poised state that allows rapid meiotic reentry in mouse GV oocytes may be determined by the differential translational control of two Cyclins.

Multifunctional Nanotube-Mucoadhesive Poly(methyl vinyl ether-co-maleic acid)@Hydroxypropyl Methylcellulose Acetate Succinate Composite for Site-Specific Oral Drug Delivery.

An advanced oral drug delivery system that can effectively deliver drugs with poor oral bioavailability is strongly desirable. Herein, a multifunctional nano-in-micro structured composite is developed by encapsulation of the mucoadhesive poly(methyl vinyl ether-co-maleic acid) modified halloysite nanotubes (HNTs) with the pH-responsive hydroxypropyl methylcellulose acetate succinate by the microfluidics to control the drug release, increase cell-particle interaction, and improve drug absorption. The microparticles show spherical shape, homogeneous particle size distribution (58 ± 1 µm), and pH-responsive dissolution behavior at pH > 6, and they prevent the premature release of curcumin in simulated pH conditions of the stomach and immediately release the curcumin in simulated pH conditions of the small intestine. The surface modification of HNT with mucoadhesive poly(methyl vinyl ether-co-maleic acid) significantly enhances its interactions with the intestinal Caco-2/HT29-MTX cells and the mouse small intestines, and increases the permeability of curcumin across the co-cultured Caco-2/HT29-MTX cell monolayers by about 13 times compared to the free curcumin. Therefore, the developed multifunctional nanotube-mucoadhesive poly(methyl vinyl ether-co-maleic acid)@hydroxypropyl methylcellulose acetate succinate composite is a promising oral drug delivery system for drugs with poor oral bioavailability.

A comprehensive review of the neonatal Fc receptor and its application in drug delivery.

Advances in the understanding of neonatal Fc receptor (FcRn) biology and function have demonstrated that this receptor, primarily identified for the transfer of passive immunity from mother infant, is involved in several biological and immunological processes. In fact, FcRn is responsible for the long half-life of IgG and albumin in the serum, by creating an intracellular protein reservoir, which is protected from lysosomal degradation and, importantly, trafficked across the cell. Such discovery has led researchers to hypothesize the role for this unique receptor in the controlled delivery of therapeutic agents. A great amount of FcRn-based strategies are already under extensive investigation, in which FcRn reveals to have profound impact on the biodistribution and half-life extension of therapeutic agents. This review summarizes the main findings on FcRn biology, function and distribution throughout different tissues, together with the main advances on the FcRn-based therapeutic opportunities and model systems, which indicate that this receptor is a potential target for therapeutic regimen modification.

Communication from the periphery to the hypothalamus through the blood-brain barrier: An in vitro platform.

One of the major routes of communication from the peripheral systems to the hypothalamus, the core structure of body homeostasis, is the humoral transmission through the blood-brain barrier (BBB). The BBB cultures are the in vitro model of choice to depict the mechanisms behind blood-brain interplay. Still, this strategy excludes the integration of the brain tissue response and, therefore, the resulting output might be limited. In this study, two in vitro assays were established: BBB coculture model and hypothalamic organotypic cultures. The combination of these two assays was used as a platform to address the two critical steps in the humoral transmission through the BBB to the brain: blood-BBB/BBB-brain. The in vitro model of the BBB was performed according to a coculture system using a brain microvascular endothelial cell line (bEnd.3) and primary astrocytes. The expression of junctional molecules as claudin-5, ZO-1, occludin and VE-cadherin was observed in the bEnd.3 cell-cell contact, confirming the BBB phenotype of these endothelial cells. Moreover, the transendothelial electrical resistance (TEER) values (71.1±9.4Ω× cm(2)) and the permeability coefficients (Pe) obtained in the transendothelial flux test (3.3±0.11×10(-6)cm/sec) support high integrity of the established barrier. The hypothalamic organotypic cultures were prepared from 8-days-old C57Bl/6 mice brains, based on the air-medium interface culture method. High cell viability (82±9.6%) and a dense neuronal network were achieved. The stimulation with dexamethasone resulted in an increased neuropeptide (NPY) expression, confirming the responsiveness of the neuronal system of these organotypic cultures. After optimization and characterization of each assay, the functionality of the platform was validated through the evaluation of the hypothalamic response to deep wound encompassing skin and muscle in mice. Results allowed to identify increased NPY activity in hypothalamic slices in response to peripheral signals within the plasma from wounded animals when compared with non-injured animals after surpassing and/or interacting with the BBB. This differential NPY response between the different animal conditions validated the functionality of the in vitro platform. In conclusion, this approach can be greatly anticipated as a useful tool for studying biologic or pharmacological circulating molecules and their impact on the hypothalamic activity.

Will dapivirine redeem the promises of anti-HIV microbicides? Overview of product design and clinical testing.

Microbicides are being developed in order to prevent sexual transmission of HIV. Dapivirine, a non-nucleoside reverse transcriptase inhibitor, is one of the leading drug candidates in the field, currently being tested in various dosage forms, namely vaginal rings, gels, and films. In particular, a ring allowing sustained drug release for 1month is in an advanced stage of clinical testing. Two parallel phase III clinical trials are underway in sub-Saharan Africa and results are expected to be released in early 2016. This article overviews the development of dapivirine and its multiple products as potential microbicides, with particular emphasis being placed on clinical evaluation. Also, critical aspects regarding regulatory approval, manufacturing, distribution, and access are discussed.