Generativity in Later-Life: The Interplay Between Retirement Status and Human, Social, and Financial Capital.

Generativity has been increasingly recognized as an important component of healthy aging. Although the desire to be generative is influenced by societal and cultural expectations, the relative influence of its driving factors by retirement status, a significant life-course transition, is underexplored. This study examined how later-life generativity is driven by the interplay between retirement status and financial, human and social capital. An online survey targeting Hong Kong adults aged 45+ was conducted. Linear regression models were stratified by retirement status (working and retired) to examine the effects of financial (income, assets, and financial satisfaction), human (education and health-related measures), and social capitals (productive and social engagement) on generativity. Among those working, higher generativity was associated with financial, human, and social capitals that facilitated material provision. Among those retired, human and social capitals that supported the transmission of knowledge and experience were more important for generativity. For both groups, support from close social networks was the strongest predictor. Different cultural demands, dictated by retirement status, play a crucial role in determining how older adults feel like they can contribute to subsequent generations. These findings can inform policies and programs that seek to support healthy transitions into retirement.

High-Throughput Cellular Heterogeneity Analysis in Cell Migration at the Single-Cell Level.

Cancer cell migration represents an essential step toward metastasis and cancer deaths. However, conventional drug discovery focuses on cytotoxic and growth-inhibiting compounds rather than inhibitors of migration. Drug screening assays generally measure the average response of many cells, masking distinct cell populations that drive metastasis and resist treatments. Here, this work presents a high-throughput microfluidic cell migration platform that coordinates robotic liquid handling and computer vision for rapidly quantifying individual cellular motility. Using this innovative technology, 172 compounds were tested and a surprisingly low correlation between migration and growth inhibition was found. Notably, many compounds were found to inhibit migration of most cells while leaving fast-moving subpopulations unaffected. This work further pinpoints synergistic drug combinations, including Bortezomib and Danirixin, to stop fast-moving cells. To explain the observed cell behaviors, single-cell morphological and molecular analysis were performed. These studies establish a novel technology to identify promising migration inhibitors for cancer treatment and relevant applications.

Later-life depressive symptoms during the Covid-19 pandemic: Investigations of individual, cumulative, and synergistic effects of social isolation.

OBJECTIVES: This study examines associations between social isolation and depressive symptoms among Hong Kong Chinese adults aged 65 and older by investigating the distinct effects of individual indicators, cumulative index, and typologies of social isolation during the Covid-19 pandemic. METHODS: We used a sample of 260 older adults from a cross-sectional, city-wide online survey targeting 1,109 aged 45+ adults through purposive sampling. Seven indicators of social isolation (not married; living alone; not engaging in social/organizational activities; no social contact with friends or families; lack of family and friends networks; loneliness) using Cornwell & Waite's framework were selected to construct three unique types of social isolation measures. We used latent class analysis (LCA) and regression models to examine the effects of varied typologies of social isolation on depressive symptoms. RESULTS: Individual model of social isolation showed that lack of social contact and feeling lonely were significant predictors of depressive symptoms. A strong linear-trend gradient effect of cumulative social isolation on depressive symptoms was also observed. The LCA model identified four typologies of social isolation (socially isolated; living alone but socially engaged; married but lacking social ties, and not socially isolated); those in the 'socially isolated' and 'married but lacking social ties' groups had the most depressive symptoms. CONCLUSION: Three operationalizations of social isolation demonstrated different utilities and implications in assessing the impacts of social isolation on depressive symptoms. Social contacts and loneliness, rather than living status or other characteristics of isolation, were the factors most strongly associated with depressive symptoms. Support programs should target lonely older adults who lack social engagement opportunities, as they are at increased risk of depression.

Developing an Operationalized Framework for Comparing Consumer-Directed Care for Older Adults: Evidence from Expert Survey and Cross-National Comparison.

Consumer-directed care (CDC) programs for older people aim to optimize health outcomes by offering clients control and flexibility regarding service arrangements. However, policy design features may differ due to heterogenous sociostructural systems. By operationalizing a framework with three dimensions of CDC, i.e. control and direct services, variety of service options, and information and support, we analyzed how countries vary in their policy designs to achieve consumer direction. Using an expert survey (n = 20) and cross-national document analysis, we analyzed 12 CDC programs from seven selected countries: the United States, the United Kingdom, Germany, the Netherlands, China, Australia, and Spain. Among the three dimensions, CDC programs placed more emphasis on and displayed more homogenous performance of policy designs that achieve consumer direction in the dimension of control and direct services, while less emphasis was placed on and more heterogenous performance displayed in the dimensions of variety of service options and information and support. We offer a systematically operationalized framework to investigate CDC policy designs. Findings advance our understanding of CDC policy features from a cross-national perspective. Policymakers could incorporate these findings to empower older people in their respective societies.

Civic Activities and Mental Health in Later Life: The Moderating Role of Generative Concerns.

This study examined the moderating mechanisms of generative concerns (perception of making contributions to others) between generative civic activities and mental health among middle-aged and older adults. A total of 1,109 community-dwelling adults aged over 45 were recruited through an online survey. Generative civic activities (political participation and volunteering), generative concerns (the Loyola Generativity Scale), and two outcomes of mental health (depressive symptoms and mental wellbeing) were measured. Linear regression models and simple slope analyses were used to probe the moderating effects of generative concerns, stratified by age (45-64 years and 65+). Generative activities, but not concerns, were associated with lower depressive symptoms among middle-aged and older adults. Generative concerns and activities were related to better mental wellbeing among middle-aged adults. Generative concerns moderated the associations between civic activities and depressive symptoms. Those with higher generative concerns but lower civic activities had higher depression scores. Conversely, a stronger reduction in depressive symptoms by engaging in civic activities was found only when individuals had greater concerns. No moderating effect was found on mental wellbeing. The positive effect of civic engagement on reducing depressive symptoms depends on generative concerns, particularly among older people and those with greater concerns. Bridging the gap between perceived generative concerns and actual civic actions may improve later-life mental health.

LANCE: a Label-Free Live Apoptotic and Necrotic Cell Explorer Using Convolutional Neural Network Image Analysis.

Identifying and quantifying cell death is the basis for all cell death research. Current methods for obtaining these quantitative measurements rely on established biomarkers, yet the marker-based approach suffers from limited marker specificity, high cost of reagents, lengthy sample preparation, and fluorescence imaging. Based on the morphological difference, we developed a Live, Apoptotic, and Necrotic Cell Explorer (LANCE) to categorize cell death status in a label-free manner, by incorporating machine learning and image processing. The LANCE workflow includes cropping individual cells from microscopic images having hundreds of cells, formation of an image database of around 5000 events, training and validation of the convolutional neural network models using multiple cell lines, and treatment conditions. With LANCE, we precisely categorized live, apoptotic, and necrotic cells with a high accuracy of 96.3 ± 0.5%. More importantly, the nondestructive label-free LANCE method allows for tracking time dynamics of the cell death process, which enhances the understanding of subtle cell death regulation at the molecular level. Hence, LANCE is a fast, low-cost, and nondestructive label-free method to distinguish cell status, which can be applied to cell death studies as well as many other biomedical applications.

The intersectionality of life course socioeconomic status, race, and cognitive decline: An 18-year follow-up.

OBJECTIVES: Studies have documented the impact of childhood socioeconomic status (SES) on cognition. However, research that simultaneously considers SES in varied life stages, the multidimensional mechanisms, and racial differences is relatively understudied. This study examines the intersectionality across age, SES, and race and its impact on cognitive trajectories. METHODS: Using 8376 respondents aged 65+ from the 1998-2016 Health and Retirement Study, we used latent growth curve modeling to examine the effects of four life course models (latency, pathway, accumulation, and mobility) on 18-year trajectories of mental status and episodic memory. We further tested for differences in the links between SES and cognitive trajectories between black and white respondents. RESULTS: Cognitive function declines with age and is interrelated with SES and race. Adulthood has a stronger effect on cognitive performance than childhood. However, linked positive childhood and adulthood SES contributes to positive cognition. Accumulated SES disadvantages were associated with lower cognition. Older adults with downward mobility and low SES throughout their lifespans had the lowest cognition scores. Life course models operated differently on trajectories of cognitive decline, yet the effects were particularly evident among older black respondents. Overall, those with socioeconomic advantages tended to have a slower decline in cognition, while a faster decline occurred for those with accrued disadvantages. CONCLUSIONS: Cognitive performance is a complex, longitudinal process intertwined with socioeconomic conditions and population heterogeneity shaped by life course contexts. Policies that facilitate healthy cognitive performance and address SES inequality could equalize health opportunities and address racial cognitive disparities later in life.

Home as the first site for suicide prevention: a Hong Kong experience.

INTRODUCTION: There has been little research into at-home suicide cases globally, and particularly in Asian regions. This study aimed to investigate the differences in characteristics between suicide cases in Hong Kong that occurred at home and elsewhere; identify at-home suicide hotspots in the community and compare the differences in area-level characteristics between suicide hotspots and other areas. METHODS: Suicide cases (2013-2017) were identified from Hong Kong Coroner's Court reports. Area-level socioeconomic data were retrieved from the 2016 Hong Kong census. Wilcoxon signed-rank tests, χ2 tests and multiple logistic regression models were applied to compare differences in characteristics between people committing suicide at home and elsewhere. Global hotspot tests (Moran's I and Getis-Ord General G) and local analysis (Getis-Ord Gi*) identified at-home suicide community hotspots. The Wilcoxon signed-rank test was used to compare differences in area-level characteristics between at-home suicide hotspots and non-hotspots. RESULTS: About 60% of suicide cases in Hong Kong occurred at home. Being female, widowed and/or living alone were significant predictors of at-home suicide cases. A U-shaped association between age and at-home suicide was identified, with 32 years of age being the critical turning point. An at-home suicide hotspot was identified in the north-western region of Hong Kong, which had lower median household income, higher income inequality and higher percentages of households with single elderly people, and new arrivals, compared with other areas. CONCLUSION: Suicide prevention should start at home by restricting access to suicide methods. Community-based suicide prevention interventions with improvement of social services should target vulnerable members in identified suicide hotspots.

Single-cell analysis of progenitor cell dynamics and lineage specification in the human fetal kidney.

The mammalian kidney develops through reciprocal interactions between the ureteric bud and the metanephric mesenchyme to give rise to the entire collecting system and the nephrons. Most of our knowledge of the developmental regulators driving this process arises from the study of gene expression and functional genetics in mice and other animal models. In order to shed light on human kidney development, we have used single-cell transcriptomics to characterize gene expression in different cell populations, and to study individual cell dynamics and lineage trajectories during development. Single-cell transcriptome analyses of 6414 cells from five individual specimens identified 11 initial clusters of specific renal cell types as defined by their gene expression profile. Further subclustering identifies progenitors, and mature and intermediate stages of differentiation for several renal lineages. Other lineages identified include mesangium, stroma, endothelial and immune cells. Novel markers for these cell types were revealed in the analysis, as were components of key signaling pathways driving renal development in animal models. Altogether, we provide a comprehensive and dynamic gene expression profile of the developing human kidney at the single-cell level.

Longitudinal impacts of grandparent caregiving on cognitive, mental, and physical health in China.

OBJECTIVES: Although studies have documented the effects of grandparenting, little is known about grandparents' long-term health outcomes in terms of optimal engagement with grandchildren, and whether age, gender, or location make a difference. This study explores the longitudinal impacts of grandparenting on health, with considerations for subgroup differences. METHODS: Using three waves of China Health and Retirement Longitudinal Study, we used propensity score analysis combined with multilevel analysis to examine the longitudinal effect of grandparenting (no, low-, moderate-, and high-intensity) on health (self-rated health, mobility limitations, depressive symptoms, and cognition) among 4,925 grandparents with at least one grandchild, and how impact varies by age, gender, and urban/rural areas. RESULTS: A nonlinear relationship between grandparenting and health was observed. Compared to those not providing care, grandparents who provided care at a low-to-moderate level were more likely to have fewer mobility limitations and depressive symptoms and better cognition. High intensity had no positive impacts on health. The effects of grandparenting on health operated differently across subgroups. Older grandparents showed better physical, mental, and cognitive health compared to their younger counterparts. Both genders had fewer mobility limitations if they provided care at a low-to-moderate level, although females reported better self-rated health. Rural grandparents reported better physical health; whereas urban grandparents reported better cognition. CONCLUSION: Grandparenting could improve health in later life; however, there are complex interactions at play. Policies aimed at supporting grandparenting should consider optimal hours of care relevant to age, gender, and urban/rural contexts.

Withholding of M-CSF Supplement Reprograms Macrophages to M2-Like via Endogenous CSF-1 Activation.

Macrophage colony-stimulating factor (M-CSF or CSF-1) is known to have a broad range of actions on myeloid cells maturation, including the regulation of macrophage differentiation, proliferation and survival. Macrophages generated by M-CSF stimulus have been proposed to be alternatively activated or M2 phenotype. M-CSF is commonly overexpressed by tumors and is also known to enhance tumor growth and aggressiveness via stimulating pro-tumor activities of tumor-associated macrophages (TAMs). Currently, inhibition of CSF-1/CSF-1R interaction by therapeutic antibody to deplete TAMs and their pro-tumor functions is becoming a prevalent strategy in cancer therapy. However, its antitumor activity shows a limited single-agent effect. Therefore, macrophages in response to M-CSF interruption are pending for further investigation. To achieve this study, bone marrow derived macrophages were generated in vitro by M-CSF stimulation for 7 days and then continuously grown until day 21 in M-CSF absence. A selective pressure for cell survival was initiated after withdrawal of M-CSF. The surviving cells were more prone to M2-like phenotype, even after receiving interleukin-4 (IL-4) stimulation. The transcriptome analysis unveiled that endogenous CSF-1 level was dramatically up-regulated and numerous genes downstream to CSF-1 covering tumor necrosis factor (TNF), ras-related protein 1 (Rap1) and phosphatidylinositol 3-kinase (PI3K)-protein kinase B (AKT) signaling pathway were significantly modulated, especially for proliferation, migration and adhesion. Moreover, the phenomenal increase of miR-21-5p and genes related to pro-tumor activity were observed in parallel. In summary, withholding of CSF-1/CSF-1R interaction would rather augment than suspend the M-CSF-driven pro-tumor activities of M2 macrophages in a long run.

Serum Angiopoietin-like Protein 3 Level Is Associated with Peripheral Arterial Stiffness in Patients with Coronary Artery Disease.

Background and Objectives: Angiopoietin-like protein 3 (ANGPTL3) is a secretory protein regulating lipid metabolism. This study evaluated the relationship between serum ANGPTL3 level and peripheral arterial stiffness (PAS) in patients with coronary artery disease (CAD). Materials and Methods: Fasting blood samples were collected from 95 CAD patients. PAS was defined as left or right brachial-ankle pulse wave velocity (baPWV) > 18.0 m/s by an oscillometric method. Serum ANGPTL3 levels were assessed using a commercial enzyme-linked immunosorbent assay kit. Results: Seventeen CAD patients (17.9%) had PAS. Patients with PAS had a significantly higher percentage of diabetes (p = 0.002), older age (p = 0.030), higher systolic blood pressure (p = 0.016), higher fasting glucose (p = 0.008), serum C-reactive protein (p = 0.002), and ANGPTL3 level (p = 0.001) than those without PAS. After multivariable logistic regression analysis, serum ANGPTL3 level (Odds ratio (OR): 1.004, 95% confidence interval (CI): 1.000-1.007, p = 0.041) is still independently associated with PAS in CAD patients. The receiver operating characteristic curve for PAS prediction revealed that the area under the curve for ANGPTL3 level was 0.757 (95% CI: 0.645-0.870, p < 0.001). Conclusions: Serum fasting ANGPTL3 level is positively associated with PAS in CAD patients. Further studies are required for clarification.

Enhanced mitochondrial fission suppresses signaling and metastasis in triple-negative breast cancer.

BACKGROUND: Mitochondrial dynamics underlies malignant transformation, cancer progression, and response to treatment. Current research presents conflicting evidence for functions of mitochondrial fission and fusion in tumor progression. Here, we investigated how mitochondrial fission and fusion states regulate underlying processes of cancer progression and metastasis in triple-negative breast cancer (TNBC). METHODS: We enforced mitochondrial fission and fusion states through chemical or genetic approaches and measured migration and invasion of TNBC cells in 2D and 3D in vitro models. We also utilized kinase translocation reporters (KTRs) to identify single cell effects of mitochondrial state on signaling cascades, PI3K/Akt/mTOR and Ras/Raf/MEK/ERK, commonly activated in TNBC. Furthermore, we determined effects of fission and fusion states on metastasis, bone destruction, and signaling in mouse models of breast cancer. RESULTS: Enforcing mitochondrial fission through chemical or genetic approaches inhibited migration, invasion, and metastasis in TNBC. Breast cancer cells with predominantly fissioned mitochondria exhibited reduced activation of Akt and ERK both in vitro and in mouse models of breast cancer. Treatment with leflunomide, a potent activator of mitochondrial fusion proteins, overcame inhibitory effects of fission on migration, signaling, and metastasis. Mining existing datasets for breast cancer revealed that increased expression of genes associated with mitochondrial fission correlated with improved survival in human breast cancer. CONCLUSIONS: In TNBC, mitochondrial fission inhibits cellular processes and signaling pathways associated with cancer progression and metastasis. These data suggest that therapies driving mitochondrial fission may benefit patients with breast cancer.

Early Prediction of Single-Cell Derived Sphere Formation Rate Using Convolutional Neural Network Image Analysis.

Functional identification of cancer stem-like cells (CSCs) is an established method to identify and study this cancer subpopulation critical for cancer progression and metastasis. The method is based on the unique capability of single CSCs to survive and grow to tumorspheres in harsh suspension culture environment. Recent advances in microfluidic technology have enabled isolating and culturing thousands of single cells on a chip. However, tumorsphere assay takes a relatively long period of time, limiting the throughput of this assay. In this work, we incorporated machine learning with single-cell analysis to expedite tumorsphere assay. We collected 1,710 single-cell events as the database and trained a convolutional neural network model that predicts whether a single cell could grow to a tumorsphere on Day 14 based on its Day 4 image. With this future-telling model, we precisely estimated the sphere formation rate of SUM159 breast cancer cells to be 17.8% based on Day 4 images. The estimation was close to the ground truth of 17.6% on Day 14. The preliminary work demonstrates not only the feasibility to significantly accelerate tumorsphere assay but also a synergistic combination between single-cell analysis with machine learning, which can be applied to many other biomedical applications.

Exhaled breath condensate based breath analyser - a disposable hydrogen peroxide sensor and smart analyser.

Exhaled breath condensate (EBC) based breath analysis has gained significant interest in pulmonary disease diagnostics over the past few years due to the non-invasiveness and simplicity of the technique. Most approaches to date have separated EBC collection from the subsequent lab-based analysis. Here we report a low-cost disposable hydrogen peroxide (H2O2)sensor modified with a new composite consisting of Prussian Blue (PB), PEDOT:PSS, and crosslinker ethylene glycol (EG) and divinyl sulfone (DVS) combined with a thermoelectric EBC collection device. Theoretical modelling suggests that previously reported large variations between analyte concentrations arise from poor control of the condensation process. Experimental data confirm the model predictions and proof-of-principle human samples were analysed showing good agreement with fluorometric methods.

Label-Free Estimation of Therapeutic Efficacy on 3D Cancer Spheres Using Convolutional Neural Network Image Analysis.

Despite recent advances in cancer treatment, developing better therapeutic reagents remains an essential task for oncologists. To accurately characterize drug efficacy, 3D cell culture holds great promise as opposed to conventional 2D monolayer culture. Due to the advantages of cell manipulation in high-throughput, various microfluidic platforms have been developed for drug screening with 3D models. However, the dissemination of microfluidic technology is overall slow, and one missing part is fast and low-cost assay readout. In this work, we developed a microfluidic chip forming 1920 tumor spheres for drug testing, and the platform is supported by automatic image collection and cropping for analysis. Using conventional LIVE/DEAD staining as the ground truth of sphere viability, we trained a convolutional neural network to estimate sphere viability based on its bright-field image. The estimated sphere viability was highly correlated with the ground truth (R-value > 0.84). In this manner, we precisely estimated drug efficacy of three chemotherapy drugs, doxorubicin, oxaliplatin, and irinotecan. We also cross-validated the trained networks of doxorubicin and oxaliplatin and found common bright-field morphological features indicating sphere viability. The discovery suggests the potential to train a generic network using some representative drugs and apply it to many different drugs in large-scale screening. The bright-field estimation of sphere viability saves LIVE/DEAD staining reagent cost and fluorescence imaging time. More importantly, the presented method allows viability estimation in a label-free and nondestructive manner. In short, with image processing and machine learning, the presented method provides a fast, low-cost, and label-free method to assess tumor sphere viability for large-scale drug screening in microfluidics.

Single-cell RNA-sequencing of migratory breast cancer cells: discovering genes associated with cancer metastasis.

Considerable evidence suggests breast cancer metastasis arises from cells undergoing epithelial-to-mesenchymal-transition (EMT) and cancer stem-like cells (CSCs). Using a microfluidic device that enriches migratory breast cancer cells with enhanced capacity for tumor formation and metastasis, we identified genes differentially expressed in migratory cells by high-throughput single-cell RNA-sequencing. Migratory cells exhibited overall signatures of EMT and CSCs with variable expression of marker genes, and they retained expression profiles of EMT over time. With single-cell resolution, we discovered intermediate EMT states and distinct epithelial and mesenchymal sub-populations of migratory cells, indicating breast cancer cells can migrate rapidly while retaining an epithelial state. Migratory cells showed differential profiles for regulators of oxidative stress, mitochondrial morphology, and the proteasome, revealing potential vulnerabilities and unexpected consequences of drugs. We also identified novel genes correlated with cell migration and outcomes in breast cancer as potential prognostic biomarkers and therapeutic targets to block migratory cells in metastasis.

Environment patterns and mental health of older adults in long-term care facilities: the role of activity profiles.

Objectives: This study adopts the International Classification of Functioning, Disability and Health (ICF) model to determine extent to which the clustered patterns of long-term care (LTC) environment and activity participation are associated with older residents' mental health. Method: This study enrolled a stratified equal probability sample of 634 older residents in 155 LTC institutions in Taiwan. Latent profile analysis and latent class analysis were conducted to explore the profiles for environment and activity participation. Multilevel modeling was performed to elucidate the hypothesized relationships. Results: Three environment profiles (Low-, Moderate-, and High-Support Environment) based on physical, social, and attitudinal environment domains and two activity profiles (Low- and High-Activity Participation) across seven activity domains were identified. Compared to the Low-Support class, older adults in the Moderate- and High-Support Environment classes had better mental health. Older residents in those two classes were more likely to be in the "High Activity Participation" class, which in turn, exhibited better mental health. Conclusion: Environment and activity participation directly relate to older residents' mental health. Activity participation also mediates the link between environment and mental health. A combination of enhanced physical, social, and attitudinal environments, and continual engagement in various activities may optimize older LTC residents' mental health.

Scalable Multiplexed Drug-Combination Screening Platforms Using 3D Microtumor Model for Precision Medicine.

Cancer heterogeneity is a notorious hallmark of this disease, and it is desirable to tailor effective treatments for each individual patient. Drug combinations have been widely accepted in cancer treatment for better therapeutic efficacy as compared to a single compound. However, experimental complexity and cost grow exponentially with more target compounds under investigation. The primary challenge remains to efficiently perform a large-scale drug combination screening using a small number of patient primary samples for testing. Here, a scalable, easy-to-use, high-throughput drug combination screening scheme is reported, which has the potential of screening all possible pairwise drug combinations for arbitrary number of drugs with multiple logarithmic mixing ratios. A "Christmas tree mixer" structure is introduced to generate a logarithmic concentration mixing ratio between drug pairs, providing a large drug concentration range for screening. A three-layer structure design and special inlets arrangement facilitate simple drug loading process. As a proof of concept, an 8-drug combination chip is implemented, which is capable of screening 172 different treatment conditions over 1032 3D cancer spheroids on a single chip. Using both cancer cell lines and patient-derived cancer cells, effective drug combination screening is demonstrated for precision medicine.