Metabolomics approach for increasing CHO cell specific productivity

Chinese hamster ovary cells are the most commonly used expression system in the production of monoclonal antibody therapeutic drugs. The biomanufacturing industry has made significant advances in increasing protein titers of these cell cultures by over 100-fold since the 1980s to gram-per-liter ranges, and much of this progress has been made via increasing cell density and viability. However, even next generation processes are approaching the limits of how high cell densities can be reached with available technologies. On the other hand, the specific productivity (qP) of the cell lines, though much higher now than at the advent of biologics production, has not been improved to the same degree, and advances on this front are needed to attain higher titers in shorter times. In this work, a library of twelve cell lines, having a wide range of qPs but all derived from the same parental cell line and expressing one of two different antibodies, was investigated using an untargeted metabolomics approach. Spent medium samples were collected from each fed-batch culture at two time points. BioCAn (Biologically Consistent Annotation), a recently developed automated annotation tool, was used to determine the most likely identities of features detected in LC-MS data from these cell lines. A correlation analysis was then performed to find annotated features that were significantly associated with either cell growth (37 features), qP (32 features), or both (56 features). Interestingly, all features associated with cell growth showed a negative correlation, while all features associated with qP showed a positive correlation. To investigate whether metabolites positively correlated with qP reflect endogenous metabolic activity beneficial for productivity, several metabolites were added to the culture medium at varying concentrations. We found that supplementing the medium with one or more select metabolites could improve qP without negatively impacting cell growth. We next evaluated whether these metabolites could be used as biomarkers to identify clones with potential for high productivity, as current screening methods can falsely eliminate clones due to sub-optimal culture media or process conditions. Together, these studies demonstrate opportunities for using untargeted metabolomics to achieve higher titer in biologics production processes. Further, the identification of biomarkers has potential to shorten cell line development timelines, which is on the critical path to biologics manufacturing

Environmental sensitivity in children: development of the highly sensitive child scale and identification of sensitivity groups

A large number of studies document that children differ in the degree they are shaped by their developmental context with some being more sensitive to environmental influences than others. Multiple theories suggest that Environmental Sensitivity is a common trait predicting the response to negative as well as positive exposures. However, most research to date relied on more or less proximal markers of Environmental Sensitivity. In this paper we introduce a new questionnaire—the Highly Sensitive Child (HSC) scale—as a promising self-report measure of Environmental Sensitivity. After describing the development of the short 12-item HSC scale for children and adolescents, we report on the psychometric properties of the scale, including confirmatory factor analysis and test-retest reliability. After considering bivariate and multivariate associations with well-established temperament and personality traits, we apply Latent Class Analysis to test for the existence of hypothesised sensitivity groups. Analyses are conducted across five studies featuring four different UK-based samples ranging in age from 8-19 years and with a total sample size of N = 3,581. Results suggest the 12-item HSC scale is a psychometrically robust measure that performs well in both children and adolescents. Besides being relatively independent from other common traits, the Latent Class Analysis suggests that there are three distinct groups with different levels of Environmental Sensitivity—low (approx. 25-35%), medium (approx. 41-47%), and high (20-35%). Finally, we provide exploratory cut-off scores for the categorisation of children into these different groups which may be useful for both researchers and practitioners

Environmental sensitivity in children: development of the highly sensitive child scale and identification of sensitivity groups

A large number of studies document that children differ in the degree they are shaped by their developmental context with some being more sensitive to environmental influences than others. Multiple theories suggest that Environmental Sensitivity is a common trait predicting the response to negative as well as positive exposures. However, most research to date relied on more or less proximal markers of Environmental Sensitivity. In this paper we introduce a new questionnaire—the Highly Sensitive Child (HSC) scale—as a promising self-report measure of Environmental Sensitivity. After describing the development of the short 12-item HSC scale for children and adolescents, we report on the psychometric properties of the scale, including confirmatory factor analysis and test-retest reliability. After considering bivariate and multivariate associations with well-established temperament and personality traits, we apply Latent Class Analysis to test for the existence of hypothesised sensitivity groups. Analyses are conducted across five studies featuring four different UK-based samples ranging in age from 8-19 years and with a total sample size of N = 3,581. Results suggest the 12-item HSC scale is a psychometrically robust measure that performs well in both children and adolescents. Besides being relatively independent from other common traits, the Latent Class Analysis suggests that there are three distinct groups with different levels of Environmental Sensitivity—low (approx. 25-35%), medium (approx. 41-47%), and high (20-35%). Finally, we provide exploratory cut-off scores for the categorisation of children into these different groups which may be useful for both researchers and practitioners

Mutual Accountability Is the Key to Equity-Oriented Systems Change: How Initiatives Can Create Durable Shifts in Policies and Practices

The COVID-19 pandemic and protests arising from police killings of Black Americans have drawn national attention to long-existent and worsening racialized gaps in health, wealth, and well-being that decades of investment and problem solving have been unable to close. Responding to amplified calls from communities and advocates for meaningful change, some philanthropic organizations are reexamining what and how they fund. We present findings from one such effort by the Robert Wood Johnson Foundation (RWJF) in partnership with the Urban Institute to assess the funder's health-promoting portfolio of investments in community development organizations and activities.This brief presents a framework for grantmakers seeking to understand why some past efforts have fallen short and how future investments might produce more equity-oriented, power-shifting systems change. Urban analyzed a portion of RWJF's portfolio consisting of 15 health-promoting programs and investments launched between 2013 and 2019 that aimed to integrate public health, health care, and community development to improve community health, well-being, and equity. As part of the assessment, we developed a guiding framework that proved critical to our inquiry. We were able to road-test the model as we synthesized insights from dozens of interviews with grantees and partners, community development intermediaries, and philanthropic leaders and staff. The mutual accountability framework allowed us to disentangle intended goals, necessary commitments, and actual results to think about the ways these three elements may—or may not be—aligned

Clinico-Neuropathological Findings in the Oldest Old from the Georgia Centenarian Study

Background: Centenarian studies are important sources for understanding of factors that contribute to longevity and healthy aging. Clinico-neuropathological finding is a key in identifying pathology and factors contributing to age-related cognitive decline and dementia in the oldest old. Objective: To characterize the cross-sectional relationship between neuropathologies and measures of premortem cognitive performance in centenarians. Methods: Data were acquired from 49 centenarians (≥98 years) from the Georgia Centenarian Study. Cognitive assessment from the time point closest to mortality was used (\u3c1 year for all subjects) and scores for cognitive domains were established. Neuropathologies [cerebral atrophy, ventricular dilation, atherosclerosis, cerebral amyloid angiopathy (CAA), Lewy bodies, hippocampal sclerosis (HS), hippocampal TDP-43 proteinopathy, neuritic plaque (NP) and neurofibrillary tangle (NFT) counts, Braak staging, and National Institute on Aging-Reagan Institute (NIARI) criteria for the neuropathological diagnosis of Alzheimer’s disease (AD)] were compared among subjects with different ratings of dementia. Linear regression was applied to evaluate the association between cognitive domain scores and neuropathologies. Results: Wide ranges of AD-type neuropathological changes were observed in both non-demented and demented subjects. Neocortical NFT and Braak staging were related to clinical dementia rating. Neocortical NFT and NP, Braak and NIARI staging, cerebral and ventricular atrophy, HS, CAA, and TDP-43 proteinopathy were differentially associated with poor performance in multiple cognitive domains and activities of daily living. Conclusion: AD-type pathology was associated with severe dementia and poor cognition but was not the only variable that explained cognitive impairment, indicating the complexity and heterogeneity of pathophysiology of dementia in the oldest old

CD11c+ Cells Are Gatekeepers for Lymphocyte Trafficking to Infiltrated Islets During Type 1 Diabetes.

Type 1 diabetes (T1D) is a T cell mediated autoimmune disease that affects more than 19 million people with incidence increasing rapidly worldwide. For T cells to effectively drive T1D, they must first traffic to the islets and extravasate through the islet vasculature. Understanding the cues that lead to T cell entry into inflamed islets is important because diagnosed T1D patients already have established immune infiltration of their islets. Here we show that CD11

GWAS of longevity in CHARGE consortium confirms APOE and FOXO3 candidacy.

To access publisher's full text version of this article, please click on the hyperlink in Additional Links field or click on the hyperlink at the top of the page marked Files. This article is open access.The genetic contribution to longevity in humans has been estimated to range from 15% to 25%. Only two genes, APOE and FOXO3, have shown association with longevity in multiple independent studies.We conducted a meta-analysis of genome-wide association studies including 6,036 longevity cases, age ≥90 years, and 3,757 controls that died between ages 55 and 80 years. We additionally attempted to replicate earlier identified single nucleotide polymorphism (SNP) associations with longevity.In our meta-analysis, we found suggestive evidence for the association of SNPs near CADM2 (odds ratio [OR] = 0.81; p value = 9.66 × 10(-7)) and GRIK2 (odds ratio = 1.24; p value = 5.09 × 10(-8)) with longevity. When attempting to replicate findings earlier identified in genome-wide association studies, only the APOE locus consistently replicated. In an additional look-up of the candidate gene FOXO3, we found that an earlier identified variant shows a highly significant association with longevity when including published data with our meta-analysis (odds ratio = 1.17; p value = 1.85×10(-10)).We did not identify new genome-wide significant associations with longevity and did not replicate earlier findings except for APOE and FOXO3. Our inability to find new associations with survival to ages ≥90 years because longevity represents multiple complex traits with heterogeneous genetic underpinnings, or alternatively, that longevity may be regulated by rare variants that are not captured by standard genome-wide genotyping and imputation of common variants.Netherlands Organisation of Scientific Research NWO Investments 175.010.2005.011 911-03-012 Research Institute for Diseases in the Elderly 014-93-015 RIDE2 Netherlands Genomics Initiative (NGI)/Netherlands Organisation for Scientific Research (NWO) 050-060-810 Erasmus Medical Center Erasmus University, Rotterdam Netherlands Organization for the Health Research and Development (ZonMw) Research Institute for Diseases in the Elderly (RIDE) Ministry of Education, Culture and Science Ministry for Health, Welfare and Sports European Commission (DG XII) Municipality of Rotterdam National Institutes of Health National Institute on Aging (NIA) R01 AG005407 R01 AR35582 R01 AR35583 R01 AR35584 R01 AG005394 R01 AG027574 R01 AG027576 AG023629 R01AG29451 U01AG009740 RC2 AG036495 RC4 AG039029 P30AG10161 R01AG17917 R01AG15819 R01AG30146 U01-AG023755 U19-AG023122 NHLBI HHSN 268201200036C HHSN268200800007C N01HC55222 N01HC85079 N01HC85080 N01HC85081 N01HC85082 N01HC85083 N01HC 85086 HL080295 HL087652 HL105756 National Institute of Neurological Disorders and Stroke (NINDS) National Center for Advancing Translational Sciences, CTSI UL1TR000124 National Institute of Diabetes and Digestive and Kidney Disease Diabetes Research Center (DRC) DK063491 National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) National Center for Research Resources (NCRR) NIH Roadmap for Medical Research U01 AR45580 U01 AR45614 U01 AR45632 U01 AR45647 U01 AR45654 U01 AR45583 U01 AG18197 U01-AG027810 UL1 RR024140 NIAMS R01-AR051124 RC2ARO58973 National Heart, Lung and Blood Institute's Framingham Heart Study N01-HC-25195 Affymetrix, Inc N02-HL-6-4278 Robert Dawson Evans Endowment of the Department of Medicine at Boston University School of Medicine Boston Medical Center National Institute of Arthritis, Musculoskeletal and Skin Diseases NIA R01 AR/AG 41398 NIH N01-AG-12100 NIA Intramural Research Program Hjartavernd (the Icelandic Heart Association) Althingi (the Icelandic Parliament) Illinois Department of Public Health Translational Genomics Research Institute Italian Ministry of Health ICS110.1/RF97.71 U.S. National Institute on Aging 263 MD 9164 263 MD 821336 Intramural Research Program of the NIH, National Institute on Aging 1R01AG028321 1R01HL09257

A genome-wide association study of aging

AbstractHuman longevity and healthy aging show moderate heritability (20%–50%). We conducted a meta-analysis of genome-wide association studies from 9 studies from the Cohorts for Heart and Aging Research in Genomic Epidemiology Consortium for 2 outcomes: (1) all-cause mortality, and (2) survival free of major disease or death. No single nucleotide polymorphism (SNP) was a genome-wide significant predictor of either outcome (p < 5 × 10−8). We found 14 independent SNPs that predicted risk of death, and 8 SNPs that predicted event-free survival (p < 10−5). These SNPs are in or near genes that are highly expressed in the brain (HECW2, HIP1, BIN2, GRIA1), genes involved in neural development and function (KCNQ4, LMO4, GRIA1, NETO1) and autophagy (ATG4C), and genes that are associated with risk of various diseases including cancer and Alzheimer's disease. In addition to considerable overlap between the traits, pathway and network analysis corroborated these findings. These findings indicate that variation in genes involved in neurological processes may be an important factor in regulating aging free of major disease and achieving longevity

Genome-wide meta-analysis of muscle weakness identifies 15 susceptibility loci in older men and women.

Low muscle strength is an important heritable indicator of poor health linked to morbidity and mortality in older people. In a genome-wide association study meta-analysis of 256,523 Europeans aged 60 years and over from 22 cohorts we identify 15 loci associated with muscle weakness (European Working Group on Sarcopenia in Older People definition: n = 48,596 cases, 18.9% of total), including 12 loci not implicated in previous analyses of continuous measures of grip strength. Loci include genes reportedly involved in autoimmune disease (HLA-DQA1 p = 4 × 10-17), arthritis (GDF5 p = 4 × 10-13), cell cycle control and cancer protection, regulation of transcription, and others involved in the development and maintenance of the musculoskeletal system. Using Mendelian randomization we report possible overlapping causal pathways, including diabetes susceptibility, haematological parameters, and the immune system. We conclude that muscle weakness in older adults has distinct mechanisms from continuous strength, including several pathways considered to be hallmarks of ageing