Aging is characterized by autophagy impairment that contributes to age-related disease aggravation. Moreover, it was described that the hypothalamus is a critical brain area for whole-body aging development and has impact on lifespan. Neuropeptide Y (NPY) is one of the major neuropeptides present in the hypothalamus, and it has been shown that, in aged animals, the hypothalamic NPY levels decrease. Because caloric restriction (CR) delays aging, at least in part, by stimulating autophagy, and also increases hypothalamic NPY levels, we hypothesized that NPY could have a relevant role on autophagy modulation in the hypothalamus. Therefore, the aim of this study was to investigate the role of NPY on autophagy in the hypothalamus. Using both hypothalamic neuronal in vitro models and mice overexpressing NPY in the hypothalamus, we observed that NPY stimulates autophagy in the hypothalamus. Mechanistically, in rodent hypothalamic neurons, NPY increases autophagy through the activation of NPY Y 1 and Y 5 receptors, and this effect is tightly associated with the concerted activation of PI3K, MEK/ERK, and PKA signaling pathways. Modulation of hypothalamic NPY levels may be considered a potential strategy to produce protective effects against hypothalamic impairments associated with age and to delay aging.ging is associated with accumulation of specific cellular proteins within neurons, a pathologic hallmark of many neurodegenerative diseases. Because average human life expectancy has increased, but also the prevalence of cognitive decline and dementia, aging research is now focused in finding strategies that increase both lifespan and healthspan.Autophagy is a highly regulated intracellular process involved in the turnover of most cellular constituents and in the maintenance of cellular homeostasis (1, 2). It is well described that basal autophagic activity decreases with age, contributing to the accumulation of altered macromolecules (3). In addition, autophagy impairment contributes to different aspects of aging phenotype and to aggravation of age-related diseases (4).Caloric restriction (CR), the reduced intake of calories without malnutrition, extends lifespan of many organisms, from yeast to mammals, and delays the progression of age-related diseases, at least in part, by stimulating autophagy (5-8). One major neuroendocrine effect of CR is the increase of neuropeptide Y (NPY) in the hypothalamus (9-12). The hypothalamus has a key role in the control of body homeostasis, neuroendocrine outputs, and feeding behavior. Recently, it was described that this brain area is critical for the development of whole-body aging and has impact on lifespan (13,14). In the hypothalamus, NPY is involved in the regulation of different physiological functions, such as regulation of food intake, body temperature, circadian rhythms, memory processing, and cognition (15-19). These diverse actions of NPY are mediated by G protein-coupled receptor subtypes named NPY Y 1 , Y 2 , Y 4 , and/or Y 5 (20, 21), all of which have been reported to be...

SummaryA 3D cell culture chip was used for high-throughput screening of a human neural progenitor cell line. The differential toxicity of 24 compounds was determined on undifferentiated and differentiating NPCs. Five compounds led to significant differences in IC50 values between undifferentiated and differentiating cultures. This platform has potential use in phenotypic screening to elucidate molecular toxicology on human stem cells.

3D suspension culture is generally considered a promising method to achieve efficient expansion and controlled differentiation of human pluripotent stem cells (hPSCs). In this work, we focused on developing an integrated culture platform for expansion and neural commitment of hPSCs into neural precursors using 3D suspension conditions and chemically-defined culture media. We evaluated different inoculation methodologies for hPSC expansion as 3D aggregates and characterized the resulting cultures in terms of aggregate size distribution. It was demonstrated that upon single-cell inoculation, after four days of culture, 3D aggregates were composed of homogenous populations of hPSC and were characterized by an average diameter of 139 ± 26 μm, which was determined to be the optimal size to initiate neural commitment. Temporal analysis revealed that upon neural specification it is possible to maximize the percentage of neural precursor cells expressing the neural markers Sox1 and Pax6 after nine days of culture. These results highlight our ability to define a robust method for production of hPSC-derived neural precursors that minimizes processing steps and that constitutes a promising alternative to the traditional planar adherent culture system due to a high potential for scaling-up.

No abstract

Due to the recent launch of new real estate investment funds for the logistical and industrial segment of the agribusiness supply chain, whose main purpose is the production, storage and processing of products, new discussions on this type of project at the real estate market, with an emphasis on product and market formatting. According to the IBGE, this year's national grain harvest will be a record in the historical series that began in the 70s, with growth of 3.5% compared to last year. Thus, the development of agribusiness in the country is remarkable. Considering the fundamental importance of understanding the agribusiness logistics supply chain, this work aims to define the format of the product (logistics warehouses for agribusiness), establishing the characteristics and guidelines necessary to serve the market and discuss real estate investment opportunities for this type of business. The work will use the methodology of bibliographic research, seeking references in the area. The research steps involve the understanding the logistics of agribusiness warehouses, delimiting the types and which agribusiness market segments will be served, topics in formatting the product: defining the characteristics and basic guidelines of the business to serve this market and guide the companies that will work with this product and seek investment opportunities in this type of opportunity (emerging real estate investment funds in this sector). As a result, this work seeks to characterize the logistic warehouses aimed at agribusiness, defining the needs to meet the sector and companies in the area, such as: minimum/maximum area, location and classification of the business, availability of land to make this product viable, types of warehouses available on the market that provide adequate storage and characterize the investment funds that exist for this type of business, among other information.

No abstract

Cell-based microarrays are valuable platforms for the study of cytotoxicity and cellular microenvironment because they enable high-throughput screening of large sets of conditions at reduced reagent consumption. However, most of the described microarray technologies have been applied to two-dimensional cultures, which do not accurately emulate the in vivo three-dimensional (3D) cell-cell and cell-extracellular matrix interactions.Herein, we describe the methodology for production of alginate- and Matrigel-based 3-D cell microarrays for the study of mouse and human pluripotent stem cells on two different chip-based platforms. We further provide protocols for on-chip proliferation/viability analysis and the assessment of protein expression by immunofluorescence.