Gene Expression Analysis of Adapted Insect Cells during Influenza VLP Production Using RNA-Sequencing

Adaptive laboratory evolution has been used to improve production of influenza hemagglutinin (HA)-displaying virus-like particles (VLPs) in insect cells. However, little is known about the underlying biological mechanisms promoting higher HA-VLP expression in such adapted cell lines. In this article, we present a study of gene expression patterns associated with high-producer insect High Five cells adapted to neutral pH, in comparison to non-adapted cells, during expression of influenza HA-VLPs. RNA-seq shows a decrease in the amount of reads mapping to host cell genomes along infection, and an increase in those mapping to baculovirus and transgenes. A total of 1742 host cell genes were found differentially expressed between adapted and non-adapted cells throughout infection, 474 of those being either up- or down-regulated at both time points evaluated (12 and 24 h post-infection). Interestingly, while host cell genes were found up- and down-regulated in an approximately 1:1 ratio, all differentially expressed baculovirus genes were found to be down-regulated in infected adapted cells. Pathway analysis of differentially expressed genes revealed enrichment of ribosome biosynthesis and carbohydrate, amino acid, and lipid metabolism. In addition, oxidative phosphorylation and protein folding, sorting and degradation pathways were also found to be overrepresented. These findings contribute to our knowledge of biological mechanisms of insect cells during baculovirus-mediated transient expression and will assist the identification of potential engineering targets to increase recombinant protein production in the future.     

Association of Food Intake with Sleep Durations in Adolescents from a Capital City in Northeastern Brazil

(1) Background: During adolescence, there are significant changes in food consumption, such as reducing the consumption of in natura or minimally processed foods and increasing the consumption of ultra-processed foods. Thus, eating habits can influence sleep duration and, consequently, affect the quality of life of young people. This study thus aims to estimate the association of consumption of in natura or minimally processed, processed, and ultra-processed foods with sleep durations in adolescents. (2) Methods: This is a cross-sectional study including 964 adolescents (18 to 19 years old) from the 1997 to 1998 birth cohort in São Luís, Maranhão. Food consumption was assessed using the food frequency questionnaire (FFQ) and stratified based on the NOVA classification. Sleep duration was verified using accelerometry in hours. The analysis of the association between the consumption of in natura or minimally processed, processedand ultra-processed foods with sleep durations in adolescents used crude and adjusted linear regression (by gender, age, skin color, education, economic class, work, consumption of alcohol, smoking, screen time, physical activity, use of illicit drugs, anxiety, depressive symptoms, and lean and fat mass). A directed acyclic graph (DAG) was used to determine the minimum set of adjustment factors. (3) Results: Of the 964 individuals evaluated, 52.0% were female. The mean sleep duration was 6 h (± 0.95). In the crude and adjusted analyses, no association was observed between food consumption according to the degree of processing and adolescent sleep durations. (4) Conclusion: There was no association between the consumption of in natura or minimally processed, processed, and ultra-processed foods with sleep durations.     

Hemotropic Mycoplasma spp. in Aquatic Mammals,Amazon Basin,Brazil

Hemotropic Mycoplasma spp. (hemoplasmas) are uncultivable bacteria that infect mammals, including humans. We detected a potentially novel hemoplasma species in blood samples from wild river dolphins in the Amazon River Basin, Brazil. Further investigation could determine pathogenicity and zoonotic potential of the detected hemoplasma.     

PROTEIN AI Advisor: A Knowledge-Based Recommendation Framework Using Expert-Validated Meals for Healthy Diets

AI-based software applications for personalized nutrition have recently gained increasing attention to help users follow a healthy lifestyle. In this paper, we present a knowledge-based recommendation framework that exploits an explicit dataset of expert-validated meals to offer highly accurate diet plans spanning across ten user groups of both healthy subjects and participants with health conditions. The proposed advisor is built on a novel architecture that includes (a) a qualitative layer for verifying ingredient appropriateness, and (b) a quantitative layer for synthesizing meal plans. The first layer is implemented as an expert system for fuzzy inference relying on an ontology of rules acquired by experts in Nutrition, while the second layer as an optimization method for generating daily meal plans based on target nutrient values and ranges. The system’s effectiveness is evaluated through extensive experiments for establishing meal and meal plan appropriateness, meal variety, as well as system capacity for recommending meal plans. Evaluations involved synthetic data, including the generation of 3000 virtual user profiles and their weekly meal plans. Results reveal a high precision and recall for recommending appropriate ingredients in most user categories, while the meal plan generator achieved a total recommendation accuracy of 92% for all nutrient recommendations.     

Fenótipo Inflamatório por Imagem de OCT Coronária: Características Específicas Entre Lesões De Novo,Hiperplasia Intimal e Neoaterosclerose Intra-Stent

BackgroundCoronary stenosis can be caused de novo atherosclerosis, in-stent restenosis, and in-stent neoatherosclerosis, three entities that develop from a diverse pathophysiological milieu.ObjectiveThis study aims to investigate, using optical coherence tomography (OCT), whether or not coronary lesions related to these processes differ in their local inflammatory profile.MethodsRetrospective analysis of patients with diagnosed or suspected coronary lesions who had undergone OCT imaging for clinical reasons. Macrophage and intra-plaque neovascularization were assessed by OCT and used as surrogates of local inflammation. A significance level of < 0.05 was adopted as statistically significant.ResultsFrom the 121 lesions, 74 were de novo, 29 were restenosis, and 18 were neoatherosclerosis. Neovascularization was found in 65.8% of de novo, 10.3% in restenosis, and 94.4% in neoatherosclerosis (p<0.01 for all). The volume of neovascularization was different among lesion types (950 vs. 0 vs. 6220, respectively [median values in 1000 x µm³/mm]; p<0.01 for all), which were significantly higher in neoatherosclerosis and lower in restenosis. The presence of macrophages differed among the lesions (95.9% in de novo vs. 6.9% in restenosis vs. 100% in neoatherosclerosis [p<0.01 for all]). Moreover, the intensity of macrophagic infiltration was different among lesion types (2.5 vs. 0.0 vs. 4.5, respectively [median values of macrophage score]; p<0.01 for all), significantly higher in neoatheroscleosis and lower in restenosis.ConclusionWhen compared using coronary OCT, de novo atherosclerosis, in-stent restenosis, and neoatherosclerosis presented markedly different inflammatory phenotypes.