Engineering Nanoscale Exosomes for Lysosomal Delivery of Bioactive Enzymes

There are at least 50 lysosomal storage diseases that are caused by accumulation of a substrate due to a defective enzyme. Although some treatment options are available, low efficacy, high cost, and immunogenicity are main challenges of current treatments. To overcome those limitations, we use cell-derived nanoparticles to deliver biologically active cargos into lysosomes. Naturally produced exosomes can deliver proteins without triggering an immune response. Additionally, exosome have an intrinsic ability to cross blood brain barrier benefiting patients at a late disease stage that affects the brain. After fusing either Gaussia luciferase (gLuc) or puromycin resistant protein (puro) onto a vesicular stomatitis virus glycoprotein (VSVG) with GFP/RFP reporters, we show that exosomes can be loaded with the recombinant protein without changing basic characteristics of exosomes. Moreover, we show that the biological activity of the proteins is retained in both the producer and recipient cells at a statistically significant level. As expected, the modified exosomes co-localize with both lysosomal and endosomal compartments indicating that they still undergo an endosomal pathway after an uptake assay. This work demonstrates that we can engineer nanoscale exosomes for delivery of therapeutic enzymes into lysosomes. The engineered vesicles have a great potential of becoming a method for enzyme delivery into patients with a lysosomal storage disease

Lady Triệu Thị Trinh Prays While Grooming Her Elephant

Lady Triệu Thị Trinh Prays While Grooming Her Elephant by Mai Nguyen D

Three Poems: Charges Against a Newborn; Unanswered; For the First Generation

This creative work features three poems: Charges Against a Newborn, Unanswered, For the First Generation

Old architectures in Thanh Ha - Bao Vinh area

VĂN HÓA - LỊCH SỬ HUẾ QUA GÓC NHÌN LÀNG XÃ PHỤ CẬN VÀ QUAN HỆ VỚI BÊN NGOÀI 　フエの文化と歴史：周辺集落と外部との関係からの視点より　Session 1: Huong Vinh Land with town-harbour Thanh Ha-Bao Vin

Covariates of turnover intentions of teleworking call center agents in Québec during the COVID-19 pandemic

Les télétravailleurs de plusieurs centres d'appels au Québec ont fourni des données des questionnaires sur leurs diverses demandes au travail (mesurées par les facteurs de stress organisationnels, la charge mentale et la charge émotionnelle), les ressources au travail (mesurées par l'indépendance au travail, la participation et les relations avec les superviseurs) ainsi que pour les mesures des résultats de la satisfaction au travail, de l'engagement organisationnel et de l’intention de quitter. Les hypothèses structurées par le modèle Job Demands-Resources ont été testées à l'aide de méthodes corrélationnelles. Comme prévu, les ressources au travail étaient liées de façon significative à la fois à la satisfaction au travail et à l'engagement organisationnel perçu par l'échantillon. Les demandes au travail prédisaient la satisfaction au travail, mais elles n'étaient pas liées à l'engagement organisationnel. Les implications théoriques et pratiques de ces résultats ont été discutées.Teleworkers from multiple call centers in Québec provided questionnaire data about their various job demands (measured by organizational stressors, mental load, and emotional load), job resources (measured by independence in the work, participation, and relationship with supervisors) as well as for outcome measures of job satisfaction, organizational commitment, and turnover intentions. Hypotheses structured by the JD-R model were tested using correlational methods. As predicted, job resources were significantly related to both job satisfaction and organizational commitment perceived by the sample. Job demands predicted job satisfaction, but they did not relate to organizational commitment. The theoretical and practical implications of these results were discussed

TRANSPLANTING COMMON LAW PRECEDENTS: AN APPROPRIATE SOLUTION FOR DEFECTS OF LEGISLATION IN VIETNAM, (part 2)

Vietnam is in the midst of legal and judicial reforms as it attempts to construct an appropriate framework for a successful market-based economy. It is increasingly likely that as its legal system has been changed to complement a market economy, an added degree of unpredictability has entered the situation, particularly due to the degree of legal transplantation involved. Vietnam has imported legal rules from the Civil Law and the Common Law, altering its legal system and blurring previously clear structural lines. Further change can be anticipated, and with due consideration, welcomed. Recently, the Vietnamese government has been planning to borrow Common Law precedents as a solution to fill the gaps in its sources of law, which have traditionally been of legislative origin. These sources include Codes, Laws, Ordinances, Decrees, Resolutions, and Circulars. Precedents and customary laws, on the other hand, have not been officially recognized. Partly due to the acceptance of a single source of law i.e. legislation, the application of law in Vietnam is critically evaluated not to be consistent, uniform and effective. Further, a lack of adequate legal interpretation, the uncertainty and the deficiency of the legislation constitute shortcomings of the legal system. Precedents are major sources of law in the Common Law system which is one of the most pre-eminent legal families in the world. As sources of law, precedents are usually considered to possess merits that make the legal system certain, consistent, fair, predictable and stable. Therefore, with those strengths, precedents can supply the certainty and stability which legislation lacks. Several factors favour the integration of precedents. Beside the need for Vietnam to counter the weakness of its legislation and the apparent strengths offered by precedents to fill legislative gaps are the readiness of Vietnam for judicial reform and the strong possibility of success of legal transplants whose success has elsewhere generally been confirmed. These provide strong motives for Vietnam to transplant precedents into its legal system. Therefore, the introduction of the Common Law precedents is completely an appropriate solution for defects of the Vietnamese legislation

Targeted delivery of lysosomal enzymes to the endocytic compartment in human cells using engineered extracellular vesicles.

Targeted delivery of lysosomal enzymes to the endocytic compartment of human cells represents a transformative technology for treating a large family of lysosomal storage diseases (LSDs). Gaucher disease is one of the most common types of LSDs caused by mutations to the lysosomal β-glucocerebrosidase (GBA). Here, we describe a genetic strategy to produce engineered exosomes loaded with GBA in two different spatial configurations for targeted delivery to the endocytic compartment of recipient cells. By fusing human GBA to an exosome-anchoring protein: vesicular stomatitis virus glycoprotein (VSVG), we demonstrate that the chimeric proteins were successfully integrated into exosomes which were secreted as extracellular vesicles (EVs) by producer cells. Isolation and molecular characterization of EVs confirmed that the fusion proteins were loaded onto exosomes without altering their surface markers, particle size or distribution. Further, enzyme-loaded exosomes/EVs added to cultured medium were taken up by recipient cells. Further, the endocytosed exosomes/EVs targeted to endocytic compartments exhibited a significant increase in GBA activity. Together, we have developed a novel method for targeting and delivery of lysosomal enzymes to their natural location: the endocytic compartment of recipient cells. Since exosomes/EVs have an intrinsic ability to cross the blood-brain-barrier, our technology may provide a new approach to treat severe types of LSDs, including Gaucher disease with neurological complications

APPLICATION OF IMAGE-BASED HIGH CONTENT ANALYSIS FOR THE SCREENING OF BIOACTIVE NATURAL PRODUCTS

Each bioactive compound induces phenotypic changes in target cells that can be made visible by labelling selected molecules of the cells with fluorescent dyes and/or directly observed under the high-throughput microscope. A comparison of the cellular phenotype induced by a compound of interest with known cellular targets allows predicting its mode of action. Over the past 15 years, high-throughput microscopy has been one of the fastest growing fields in cell biology. When combined with automated multiparametric image and data analysis, it is referred to as high-content screening (HCS). Whilst HCS has been successfully applied to the bioactivity characterization of natural products, recent studies used automated microscopy and software to increase speed and to reduce subjective interpretation. In 2017, Institute of Natural Products Chemistry (INPC-VAST) has been equipped with a HCS platform (Olympus Scan^R) that designed for fully automated image acquisition and analysis of biological samples to visually inspect the cellular morphology induced by hit compounds as well as to discriminate from false positives. Accordingly, this short review covers the concepts of HCS and its application in screening of biologically active natural products whose molecular targets could be identified through such approaches