Multiplexed Quantitative Proteomic Analysis of HEK293 Provides Insights into Molecular Changes Associated with the Cell Density Effect, Transient Transfection, and Virus-Like Particle Production

The production of virus-like particles (VLPs) has gained importance over the last few years owing to the benefits they provide compared to conventional vaccines. The biopharmaceutical industry is currently searching for safer candidates based on VLPs for new and existing vaccines and implementing new methods of manufacturing, thus allowing a more sustainable, effective, and species-specific production. Despite achieving lower yields compared to traditional platforms, the use of mammalian cells provides the right post-translational modifications, and consequently, the intensification of bioprocesses using mammalian cell platforms has become a matter of pressing concern. One of the methods subjected to intensification is transient gene expression, which has been proven to be highly effective regarding VLP production for preclinical or even clinical trials. In this work, a multiplexed quantitative proteomic approach has been applied to study the molecular characteristics of HEK293 cell cultures when growing at cell densities higher than 4 × 106 cells/mL and to study the effects related to cell transfection and VLP production. The obtained results revealed a set of functional and metabolic profiles of HEK293 under these three different conditions that allowed the identification of physiological bottlenecks regarding VLP production. Regarding the cell density effect, molecular alterations in the cell biology were proposed to help explain the difficulty for the cells to be transfected at higher densities. In addition, an overall disruption of cellular homeostasis after transfection was observed based on altered biological processes, and after identifying potential pathways liable to be optimized via metabolic engineering, different solutions were proposed to improve VLP production.The project that gave rise to these results received the support of a fellowship from ”la Caixa” Foundation (ID 100010434). The fellowship code is LCF/BQ/ES17/11600003. This study was supported by competitive grants from the Spanish Ministry of Economy and Competitiveness (MINECO) (BIO2015-67580-P, PGC2018-097019-BI00) through the Carlos III Institute of Health-Fondo de Investigación Sanitaria (PRB3, PT17/0019/0003 ISCIIISGEFI/FEDER) and by CIBERCV (CB16/11/00277). The CNIC is supported by the Ministerio de Ciencia, Innovación y Universidades and the Pro-CNIC Foundation, and is a Severo Ochoa Center of Excellence (SEV-2015-0505).S

Nursing Genetic Research: New Insights Linking Breast Cancer Genetics and Bone Density

Nursing research is expected to provide options for the primary prevention of disease and health promotion, regardless of pathology or disease. Nurses have the skills to develop and lead research that addresses the relationship between genetic factors and health. Increasing genetic knowledge and research capacity through interdisciplinary cooperation as well as the development of research resources, will accelerate the rate at which nurses contribute to the knowledge about genetics and health. There are currently different fields in which knowledge can be expanded by research developed from the nursing field. Here, we present an emerging field of research in which it is hypothesized that genetics may affect bone metabolism. Better insight of genetic factors that are contributing to metabolic bone diseases would allow for focused nursing care and preventive interventions

Depressed Mood after Stroke: Predictive Factors at Six Months Follow-Up

We aimed to know the prevalence of post-stroke depression (PSD) in our context, identify the variables that could predict post-stroke depression, by using the Hamilton Depression Rating Scale, occurring within six months after stroke, and identify patients at high risk for PSD. Methods: descriptive, cross-sectional and observational study. We included 173 patients with stroke (transient ischemic attack (TIA) included) and collected sociodemographic and clinical variables. We used the Hamilton Depression Scale (HDS) for depression assessment and Barthel Index and modified Rankin Scale (mRS) for functional assessment. The neurological severity was evaluated by the National Institutes of Health Stroke Scale (NIHSS). Results: 35.5% were women, aged 71.16 (±12.3). Depression was present in 42.2% patients (n = 73) at six months after stroke. The following variables were significantly associated with PSD: diagnosis of previous depression (p = 0.005), the modified Rankin Scale at discharge (p = 0.032) and length of hospital stay (p = 0.012). Conclusion: PSD is highly prevalent after stroke and is associated with the severity, left location of the stroke, and the degree of disability at discharge. Its impact justifies the evaluation and early treatment that still continues to be a challenge today

Partial correlations between LCO3-PUFAs intake and bone mineral density in normal women according to the WHO diagnosis criteria for osteoporosis.

<p>Partial correlations between LCO3-PUFAs intake and bone mineral density in normal women according to the WHO diagnosis criteria for osteoporosis.</p

Multiple linear regression analysis for the association between BMD at the femoral neck and age, BMI, calcium intake, vitamin D intake, menopausal status and total LCO3-PUFA intake according to the WHO diagnosis criteria for osteoporosis.

<p>Multiple linear regression analysis for the association between BMD at the femoral neck and age, BMI, calcium intake, vitamin D intake, menopausal status and total LCO3-PUFA intake according to the WHO diagnosis criteria for osteoporosis.</p

Bivariate and partial correlations between LCO3-PUFAs and bone mineral density.

<p>Bivariate and partial correlations between LCO3-PUFAs and bone mineral density.</p

Dietary intake of EPA and DHA according to the WHO diagnosis criteria for osteoporosis.

<p>Bars represents mean±C.I 95%. Significant differences were found for the EPA and DHA intake between normal vs osteoporosis and for osteopenia vs osteoporosis groups; p-values after further adjustment by energy intake (kcal/day) (ANCOVA test).</p

Bone mineral density of premenopausal and postmenopausal subjects.

<p>Bone mineral density of premenopausal and postmenopausal subjects.</p