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Human Embryonic Kidney cells (HEK293) are a popular host for recombinant protein expression and production in the biotechnological industry. This has driven within both, the scientific and the engineering communities, the search for strategies to increase their protein productivity. The present work is inserted into this search exploring the impact of adding sodium acetate (NaAc) into a batch culture of HEK293 cells. We monitored, as a function of time, the cell density, many external metabolites, and the supernatant concentration of the heterologous extra-cellular domain ECD-Her1 protein, a protein used to produce a candidate prostate cancer vaccine. We observed that by adding different concentrations of NaAc (0, 4, 6 and 8 mM), the production of ECD-Her1 protein increases consistently with increasing concentration, whereas the carrying capacity of the medium decreases. To understand these results we exploited a combination of experimental and computational techniques. Metabolic Flux Analysis (MFA) was used to infer intracellular metabolic fluxes from the concentration of external metabolites. Moreover, we measured independently the extracellular acidification rate and oxygen consumption rate of the cells. Both approaches support the idea that the addition of NaAc to the culture has a significant impact on the metabolism of the HEK293 cells and that, if properly tuned, enhances the productivity of the heterologous ECD-Her1 protein.

Habitual consumption of highly palatable foods when not in metabolic need (HPF eating) is linked to obesity. High HPF consumption is also linked to mental health disorder (MHD) symptoms. Mindfulness-based interventions are popular treatments for obesity and MHDs, but little is known about the relationship between trait mindfulness and motive-based HPF eating. Therefore, a total of 927 young adults completed a survey that included the Palatable Eating Motives Scale-7 (which identifies Coping-, Reward enhancement-, Social-, and Conformity-eating), the Mindful Attention Awareness Scale, the Perceived Stress Scale, and demographic and body mass index (BMI) questions. An MHD questionnaire allowed a comparison of HPF eating between participants with and without various MHDs. Regressions revealed that Coping-eating was independently associated with lower mindfulness and also greater perceived stress, higher BMI, and female sex. Of these variables, only lower mindfulness was independently associated with Reward-, Social-, and Conformity-eating. Coping- and Reward-eating were more frequent in participants with versus without an anxiety disorder, depression, ADD/ADHD, and PTSD. Coping-eating was also more frequent in participants with body dysmorphic disorder. These findings warrant investigations in participants with clinically validated diagnoses for DSM-specific MHDs. Results from such investigations and the uncovered nature of associations between motive-specific HPF eating and trait mindfulness could provide novel targets to improve mindfulness-based interventions for obesity and MHDs.

We have developed a single process for producing two key COVID-19 vaccine antigens: SARS-CoV-2 receptor binding domain (RBD) monomer and dimer. These antigens are featured in various COVID-19 vaccine formats, including SOBERANA 01 and the licensed SOBERANA 02, and SOBERANA Plus. Our approach involves expressing RBD (319-541)-His6 in Chinese hamster ovary (CHO)-K1 cells, generating and characterizing oligoclones, and selecting the best RBD-producing clones. Critical parameters such as copper supplementation in the culture medium and cell viability influenced the yield of RBD dimer. The purification of RBD involved standard immobilized metal ion affinity chromatography (IMAC), ion exchange chromatography, and size exclusion chromatography. Our findings suggest that copper can improve IMAC performance. Efficient RBD production was achieved using small-scale bioreactor cell culture (2 L). The two RBD forms - monomeric and dimeric RBD - were also produced on a large scale (500 L). This study represents the first large-scale application of perfusion culture for the production of RBD antigens. We conducted a thorough analysis of the purified RBD antigens, which encompassed primary structure, protein integrity, N-glycosylation, size, purity, secondary and tertiary structures, isoform composition, hydrophobicity, and long-term stability. Additionally, we investigated RBD-ACE2 interactions, in vitro ACE2 recognition of RBD, and the immunogenicity of RBD antigens in mice. We have determined that both the monomeric and dimeric RBD antigens possess the necessary quality attributes for vaccine production. By enabling the customizable production of both RBD forms, this unified manufacturing process provides the required flexibility to adapt rapidly to the ever-changing demands of emerging SARS-CoV-2 variants and different COVID-19 vaccine platforms.

Highly palatable foods play a salient role in obesity and binge-eating, and if habitually eaten to deal with intrinsic and extrinsic factors unrelated to metabolic need, may compromise adaptive coping and interpersonal skills. This study used event sampling methodology (ESM) to examine whether individuals who report eating palatable foods primarily to cope, to enhance reward, to be social, or to conform, as measured by the Palatable Eating Motives Scale (PEMS), actually eat these foods primarily for the motive(s) they report on the PEMS. Secondly this study examined if the previously reported ability of the PEMS Coping motive to predict BMI would replicate if the real-time (ESM-reported) coping motive was used to predict BMI. A total of 1691 palatable eating events were collected from 169 college students over 4 days. Each event included the day, time, and types of tasty foods or drinks consumed followed by a survey that included an abbreviated version of the PEMS, hunger as an additional possible motive, and a question assessing general perceived stress during the eating event. Two-level mixed modeling confirmed that ESM-reported motives correlated most strongly with their respective PEMS motives and that all were negatively associated with eating for hunger. While stress surrounding the eating event was strongly associated with the ESM-coping motive, its inclusion in the model as a predictor of this motive did not abolish the significant association between ESM and PEMS Coping scores. Regression models confirmed that scores on the ESM-coping motive predicted BMI. These findings provide ecological validity for the PEMS to identify true-to-life motives for consuming palatable foods. This further adds to the utility of the PEMS in individualizing, and hence improving, treatment strategies for obesity, binge-eating, dietary nutrition, coping, reward acquisition, and psychosocial skills.