Comparative study of using different yeast genera as vehicles for protein delivery to antigen-presenting cells

The use of yeasts as vehicle for protein antigens has been demonstrated to be a highly effective vaccination approach. In part, this can be attributed to the intrinsic adjuvant properties of yeast cell wall components. Moreover, the correct processing of recombinantly expressed proteins and the safety status of many yeast genera has encouraged the onset of preclinical and clinical trials using yeast vectors. However, the vast majority of such studies focused the attention on yeasts of the genus Saccharomyces as candidate T cell vaccine. In this work, different yeast genera were evaluated as potential antigen carrier in view of the development of novel yeast-based vaccines. For this purpose, yeasts were initially assessed for their ability to induce maturation and activation of human dendritic cells. Next, the internalization profile of selected yeasts by mammalian phagocytes was analyzed, as well as the involvement of pattern recognition receptors in the uptake process. Subsequently, yeasts engineered to express foreign proteins were assessed for their antigen delivery capacity. In vitro antigen presentation and ex vivo whole blood assays showed that recombinant yeast genera differently activate antigen-specific T cells. Furthermore, antigen localization played a decisive role in T cell activation. The data presented here strongly support the potential of recombinant yeast in the development of novel vaccine strategies in order to induce antigen-specific T cell responses.Der Einsatz von Hefen als Vehikel für Proteinantigene stellt eine vielversprechende Vakzinierungsstrategie dar, was u.a. auf adjuvante Eigenschaften der Hefe-Zellwandkomponenten zurückzuführen ist. Weiterhin haben der Nachweis der korrekten Prozessierung rekombinanter Proteine und der unbedenkliche Status vieler Hefegattungen ihren Einsatz in präklinischen und klinischen Studien gefördert. Bislang hat sich die Mehrzahl dieser Studien auf Hefen der Gattung Saccharomyces als Vakzinkandidaten für zellvermittelte Immunantworten konzentriert. Im Rahmen dieser Arbeit wurden verschiedene Hefegattungen als potentielle Antigenvehikel zur Etablierung neuartiger Hefe-basierter Vakzinen untersucht. Zunächst wurden Ausreifung und Aktivierung von Dendritischen Zellen durch diverse Hefegattungen analysiert. Danach wurden sowohl die Aufnahme bestimmter Hefegattungen durch Säuger-Phagozyten als auch die Beteiligung spezifischer Rezeptoren in diesem Prozess untersucht. Anschließend wurde die Fähigkeit rekombinanter Hefen zum Antigen "Delivery" evaluiert. Durch in vitro Antigenpräsentation und ex vivo Vollblut-Assays konnte gezeigt werden, dass verschiedene Hefegattungen Antigen-spezifische T-Zellen unterschiedlich aktivieren. Des Weiteren spielt die Antigenlokalisierung eine wichtige Rolle bei der T-Zellaktivierung. Die vorliegenden Ergebnisse unterstreichen das Potenzial rekombinanter Hefen bei der Entwicklung neuartiger Impfstrategien zur Induktion Antigen-spezifischer T-Zell-Immunantworten

Decreased resistin plasmatic concentrations in patients with Alzheimer's disease: A case-control study

Previous studies suggested a role for adipokines in ageing and in several age-related diseases. The purpose of our study was to further elucidate adipokines involvement in neurodegeneration, investigating adiponectin, leptin and resistin in Alzheimer's disease (AD) and Frontotemporal Dementia (FTD). We enrolled for the study 70 subjects: 26 AD, 21 FTD, and 23 with other neurological (but not neurodegenerative) conditions (CTR, control group). According to a standardized protocol, we measured adipokines plasmatic levels, blood parameters of glucidic and lipidic metabolism, ESR, cerebrospinal fluid (CSF) markers of neurodegeneration (beta-amyloid, total-Tau, phosphorylated-Tau) and anthropometric parameters. In comparison with control group, we found lower resistin concentrations in patients with dementia, and in particular in AD (p < 0.001). In multivariate analysis, AD relative risk was reduced by resistin, when controlling for sex, age and anthropometric/metabolic parameters (RR = 0.71, P < 0.0001). Considering CSF biomarkers, we found a direct correlation between resistin and Aβ(1-42) CSF concentration in patients (p < 0.001, r = 0.50). Lower resistin characterized AD patients in our study and AD, but not FTD, diagnosis risk was found to be inversely associated with resistin when controlling for confounders. We hypothesize that resistin-linked metabolic profile has to be reconsidered and further investigated in AD

Postoperative outcomes following rehabilitation in patients with left ventricular assist devices

Postoperative rehabilitation is a cornerstone of the recovery pathway following left ventricular assist device implantation (LVAD), and patients are expected to conduct an autonomous life thanks to improved technology and increased knowledge of mechanical circulatory support. The primary purpose of the present study was to quantify clinical changes related to rehabilitation, in patients with LVAD: functional capacity, disability, and quality of life were identified as reliable outcomes to detect such changes. The current study was a scoping review conducted searching three primary databases, namely PubMed, Scopus, and Cochrane Library, from their inception until January 2020. After the selection process was completed, 12 citations were included in the present study. Three hundred eight three patients were included in the current analysis. Functional capacity, disability, and quality of life were investigated in 157, 215, 18 patients, respectively. Significant differences were found before and after rehabilitation. The mean walked distance at 6-Minute Walk Test improved from 319±96 to 412.8±86.2 metres (p<0.001), the mean score of the Functional Independence Measure from 68.4±11.8 to 92.5±10.8 points (p<0.001), the mean score of the Short Form-36 physical component from 32.7±29.9 to 55.5±24.7 points (p=0.009) and the mental component from 55.8±19.8 to 75.4±21.4 points (p=0.002). Postoperative rehabilitation is effective at improving functional capacity, disability, and quality of life in patients with left ventricular assist device; all these three domains are particularly expressive of the entity of patients' functional recovery

Magnetic nanoparticles in the central nervous system: Targeting principles, applications and safety issues

One of the most challenging goals in pharmacological research is overcoming the Blood Brain Barrier (BBB) to deliver drugs to the Central Nervous System (CNS). The use of physical means, such as steady and alternating magnetic fields to drive nanocarriers with proper magnetic characteristics may prove to be a useful strategy. The present review aims at providing an up-to-date picture of the applications of magnetic-driven nanotheranostics agents to the CNS. Although well consolidated on physical ground, some of the techniques described herein are still under investigation on in vitro or in silico models, while others have already entered in—or are close to—clinical validation. The review provides a concise overview of the physical principles underlying the behavior of magnetic nanoparticles (MNPs) interacting with an external magnetic field. Thereafter we describe the physiological pathways by which a substance can reach the brain from the bloodstream and then we focus on those MNP applications that aim at a nondestructive crossing of the BBB such as static magnetic fields to facilitate the passage of drugs and alternating magnetic fields to increment BBB permeability by magnetic heating. In conclusion, we briefly cite the most notable biomedical applications of MNPs and some relevant remarks about their safety and potential toxicity

Investigating p62 Concentrations in Cerebrospinal Fluid of Patients with Dementia: A Potential Autophagy Biomarker In Vivo?

Several studies have revealed defects in autophagy in neurodegenerative disorders including Alzheimer&rsquo;s disease (AD) and frontotemporal dementia (FTD). SQSTM1/p62 plays a key role in the autophagic machinery and may serve as a marker for autophagic flux in vivo. We investigated the role of p62 in neurodegeneration, analyzing its concentrations in the CSF of AD and FTD patients. We recruited 76 participants: 22 patients with AD, 28 patients with FTD, and 26 controls. CSF p62 concentrations were significantly increased in AD and FTD patients when compared to controls, which persisted after adjusting for age (p = 0.01 and p = 0.008, respectively). In female FTD patients, p62 positively correlated with the neurodegenerative biomarkers t-Tau and p-Tau. A significant correlation between CSF p62 concentrations and several clinical features of AD was found. Our data show that p62 is increased in CSF of AD and FTD patients, suggesting a key role of autophagy in these two disorders. The levels of p62 in CSF may reflect an altered autophagic flux, and p62 could represent a potential biomarker of neurodegeneration

L-Carnitine Functionalization to Increase Skeletal Muscle Tropism of PLGA Nanoparticles

Muscular dystrophies are a group of rare genetic pathologies, encompassing a variety of clinical phenotypes and mechanisms of disease. Several compounds have been proposed to treat compromised muscles, but it is known that pharmacokinetics and pharmacodynamics problems could occur. To solve these issues, it has been suggested that nanocarriers could be used to allow controlled and targeted drug release. Therefore, the aim of this study was to prepare actively targeted poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) for the treatment of muscular pathologies. By taking advantage of the high affinity for carnitine of skeletal muscle cells due to the expression of Na+-coupled carnitine transporter (OCTN), NPs have been actively targeted via association to an amphiphilic derivative of L-carnitine. Furthermore, pentamidine, an old drug repurposed for its positive effects on myotonic dystrophy type I, was incorporated into NPs. We obtained monodispersed targeted NPs, with a mean diameter of about 100 nm and a negative zeta potential. To assess the targeting ability of the NPs, cell uptake studies were performed on C2C12 myoblasts and myotubes using confocal and transmission electron microscopy. The results showed an increased uptake of carnitine-functionalized NPs compared to nontargeted carriers in myotubes, which was probably due to the interaction with OCTN receptors occurring in large amounts in these differentiated muscle cells

DESENVOLVIMENTO E VALIDAÇÃO DE UM DISPOSITIVO PARA MENSURAÇÃO DA FLEXIBILIDADE POR MEIO DO ACELERÔMETRO EM CRIANÇAS

The objectives of this study were to develop and validate a computerized device to measure joint flexibility; to measure flexibility in children in elementary school aged 8 to 10 years using the Wells bank; to compare the data obtained by the Wells bank and the computerized device developed in the present study and evaluate the usability of the device. 215 volunteers participated in this research, aged between 8 and 10 years, 63.72% (137) were female and 36.28% (78) were male, duly enrolled in an educational institution in the Alto Tietê region. For data collection, the volunteers performed the flexibility test using the Wells bank and the computerized device developed in this research to assess flexibility. The computerized device it was made on a 3D printing technology printer. A usability evaluation was carried out with 15 Physical Education teachers for the product developed through a usability questionnaire based on the System Usability Scale (SUS) form, which contains 10 multiple-choice questions. The data obtained by the objective measures determined were evaluated in terms of frequency, mean and standard deviation. The arithmetic means of the three measurements performed at each stage were calculated. Mann-Whitney and Kruskal-Wallis tests and Spearman's correlation were performed for the investigation. For all analyses, significant differences were considered p&lt;0.05. It is concluded that the computerized device to measure joint flexibility is capable of evaluating the range of motion in degrees, and was qualified as easy to handle by professionals in the field of physical education.Los objetivos de este estudio fueron desarrollar y validar un dispositivo computarizado para medir la flexibilidad articular; medir la flexibilidad en niños en la escuela primaria de 8 a 10 años a través del banco Wells; comparar los datos obtenidos por la base de datos Wells y el dispositivo computarizado desarrollado en el presente estudio y evaluar la usabilidad del dispositivo. Un total de 215 voluntarios participaron en esta investigación, con edades de 8 a 10 años, 63,72% (137) mujeres y 36,28% (78) hombres, debidamente matriculados en una institución educativa de la región del Alto Tietê. Para la recolección de datos, los voluntarios realizaron la prueba de flexibilidad utilizando la base de datos Wells y el dispositivo computarizado desarrollado en esta investigación para evaluar la flexibilidad. El dispositivo fue hecho de una impresora de tecnología de impresión 3D. Se realizó una evaluación de usabilidad con 15 profesores de Educación Física para el producto desarrollado a través del cuestionario de usabilidad basado en el formulario System Usability Scale (SUS) que contiene 10 preguntas de opción múltiple. Los datos obtenidos por las mediciones objetivas determinadas fueron evaluados para la frecuencia, media y desviación estándar. Se calcularon las medias aritméticas de las tres mediciones realizadas en cada etapa. Las pruebas de Mann-Whitney y Kruskal-Wallis y la correlación de Spearman se realizaron para la investigación. Para todos los análisis, se consideraron diferencias significativas p&lt;0,05. Se concluye que el dispositivo computarizado para medir la flexibilidad articular es capaz de evaluar el rango de movimiento en grados, y fue calificado fácil de manejar por profesionales de la educación física.Os objetivos deste estudo foram desenvolver e validar um dispositivo computadorizado para mensurar a flexibilidade articular; mensurar a flexibilidade em crianças no ensino fundamental de 8 a 10 anos por meio do banco de Wells; comparar os dados alcançados pelo banco de Wells e o dispositivo computadorizado desenvolvido no presente estudo e avaliar a usabilidade do dispositivo. Participaram desta pesquisa 215 voluntários, com idade de 8 a 10 anos, sendo 63,72% (137) do sexo feminino e 36,28% (78) do sexo masculino, devidamente matriculados em uma instituição de ensino na região do Alto Tietê. Para coleta de dados, os voluntários realizaram o teste de flexibilidade utilizando o banco de Wells e o dispositivo computadorizado desenvolvido nessa pesquisa para avaliar a flexibilidade.O dispositivo foi confeccionado numa impressora de tecnologia de impressão 3D. Foi realizada uma avaliação da usabilidade com 15 professores de Educação Física para o produto desenvolvido através da interação de usabilidade baseada no formulário System Sustainability Scale (SUS) que contém 10 questões de múltipla-escolha. Os dados obtidos por medidas objetivas determinadas foram avaliadas quanto à frequência, média e desvio padrão. Foram seguidas como médias aritméticas das três medidas realizadas em cada etapa. Para investigar realizou-se os testes de Mann-Whitney e de Kruskal-Wallis e a tradução de Spearman. Para todas as análises foram consideradas diferenças significativas p&lt;0,05.Conclui-se que o dispositivo computadorizado para medir a flexibilidade articular é capaz de avaliar em graus a amplitude de movimento