Cooperative Response to Endocardial Notch Reveals Interaction With Hippo Pathway.

BACKGROUND: The endocardium is a crucial signaling center for cardiac valve development and maturation. Genetic analysis has identified several human endocardial genes whose inactivation leads to bicuspid aortic valve formation and calcific aortic valve disease, but knowledge is very limited about the role played in valve development and disease by noncoding endocardial regulatory regions and upstream factors. METHODS: We manipulated Notch signaling in mouse embryonic endocardial cells by short-term and long-term coculture with OP9 stromal cells expressing Notch ligands and inhibition of Notch activity. We examined the transcriptional profile and chromatin accessibility landscape for each condition, integrated transcriptomic, transcription factor occupancy, chromatin accessibility, and proteomic datasets. We generated in vitro and in vivo models with CRISPR-Cas9-edited deletions of various noncoding regulatory elements and validated their regulatory potential. RESULTS: We identified primary and secondary transcriptional responses to Notch ligands in the mouse embryonic endocardium, and a NOTCH-dependent transcriptional signature in valve development and disease. By defining the changes in the chromatin accessibility landscape and integrating with the landscape in developing mouse endocardium and adult human valves, we identify potential noncoding regulatory elements, validated selected candidates, propose interacting cofactors, and define the timeframe of their regulatory activity. Additionally, we found cooperative transcriptional repression with Hippo pathway by inhibiting nuclear Yap (Yes-associated protein) activity in the endocardium during cardiac valve development. CONCLUSIONS: Sequential Notch-dependent transcriptional regulation in the embryonic endocardium involves multiple factors. Notch activates certain noncoding elements through these factors and simultaneously suppresses elements that could hinder cardiac valve development and homeostasis. Biorxviv: https://www.biorxiv.org/content/10.1101/2023.03.23.533882v1.full.

Structural Factors Contributing to Compassion Fatigue, Burnout, and Secondary Traumatic Stress Among Hospital-Based Healthcare Professionals During the COVID-19 Pandemic.

High levels of burnout among healthcare providers (HCPs) have been a widely documented phenomenon, which have been exacerbated during the COVID-19 pandemic. In the United States, qualitative studies that are inclusive of HCPs in diverse professional roles have been limited. Therefore, we utilized a qualitative-quantitative design to examine professional quality of life in terms of compassion fatigue, burnout, and secondary traumatic stress among hospital-based HCPs, including social workers, hospitalists, residents, and palliative care team members during COVID-19. HCPs (n = 26) participated in virtual semi-structured focus groups or individual interviews and online surveys (n = 30) including the Professional Quality of Life (ProQOL) Scale. While ProQOL scores indicated low levels of compassion fatigue, burnout, and secondary traumatic stress, thematic analysis of our qualitative data included rich descriptions of compassion fatigue, burnout, and secondary traumatic stress. Safety concerns and value misalignment characterized structural stressors perceived to contribute to HCP compassion fatigue, burnout, and secondary traumatic stress. The discrepancy between our qualitative and quantitative findings may be indication that modifications to current screenings are warranted. These findings also suggest a need to identify and implement structural and policy changes that increase HCPs' physical and emotional safety and promote better alignment of institutional interests with HCP values.

ABC-GOALScl score predicts admission to the intensive care unit and mortality of COVID-19 patients over 60 years of age.

BACKGROUND: One of the risk factors for getting seriously ill from COVID-19 and reaching high mortality rates is older age. Older age is also associated with comorbidities, which are risk factors for severe COVID-19 infection. Among the tools that have been evaluated to predict intensive care unit (ICU) admission and mortality is ABC-GOALScl. AIM: In the present study we validated the utility of ABC-GOALScl to predict in-hospital mortality in subjects over 60 years of age who were positive for SARS-CoV-2 virus at the moment of admission with the purpose of optimizing sanitary resources and offering personalized treatment for these patients. METHODS: This was an observational, descriptive, transversal, non-interventional and retrospective study of subjects (≥ 60 years of age), hospitalized due to COVID-19 infection at a general hospital in northeastern Mexico. A logistical regression model was used for data analysis. RESULTS: Two hundred forty-three subjects were included in the study, whom 145 (59.7%) passed away, while 98 (40.3%) were discharged. Average age was 71, and 57.6% were male. The prediction model ABC-GOALScl included sex, body mass index, Charlson comorbidity index, dyspnea, arterial pressure, respiratory frequency, SpFi coefficient (Saturation of oxygen/Fraction of inspired oxygen ratio), serum levels of glucose, albumin, and lactate dehydrogenase; all were measured at the moment of admission. The area under the curve for the scale with respect to the variable of discharge due to death was 0.73 (IC 95% = 0.662-0.792). CONCLUSION: The ABC-GOALScl scale to predict ICU admission in COVID-19 patients is also useful to predict in-hospital death in COVID-19 patients ≥ 60 years old.

Suicide Within 72 Hours After Discharge From Health Care Settings: Decedent Characteristics.

Health care providers have the challenge of identifying patients at risk of committing suicide after discharge from their care. The aim of this study was to identify and describe the population committing suicide less than 72 hours after discharge from medical care. Between 2006 and 2014 in Harris County, Texas, 30 individuals were identified who met these criteria. The decedents included 27 men and 3 women with a mean age of 43.5 years. The cause of death in most cases was gunshot wound of the head. Five of the decedents had requested discharge or left against medical advice and 24 committed suicide within 24 hours. Although the sample size is small, it is one of the largest cohorts of its type.

The Value of Subjective Olfactometry as a Predictive Biomarker of Neurodegenerative Diseases: A Systematic Review.

BACKGROUND: Olfactory disorders (ODs) are reported to be an early non-motor sign before the onset of deterioration in neurodegenerative diseases (NDs) such as Alzheimer's and Parkinson's. This systematic revision aims to review the current literature and the value of subjective olfactometry (SO) in the early diagnosis of cognitive decline and NDs. METHODS: A systematic literature review was conducted following the PRISMA framework. Four different authors reviewed six different databases. The main variables analyzed were olfactory function and cognitive status. The quality of results was evaluated using the Oxford Centre of Evidence-based Medicine Levels. RESULTS: Twenty-one cross-sectional and cohort studies and six meta-analyses were included. Most of them found an association between ODs and NDs. A prevalence of ODs greater than 80% was shown in Parkinson's disease, proportional to the severity of symptoms. In Alzheimer's, ODs were associated with early diagnosis and prognosis. All SO tests employed in the literature showed enough predictive value to correlate with early stages of cognitive decline. CONCLUSIONS: SO should be considered a pivotal tool when diagnosing NDs due to their association with early symptoms and prognosis. However, in the current literature, no firm consensus exists on the optimal SO tests and protocols that should be applied to the study of NDs, which prevents the interpretability and comparability of results among studies.

Culture of cancer spheroids and evaluation of anti-cancer drugs in 3D-printed miniaturized continuous stirred tank reactors (mCSTRs).

Cancer continues to be a leading cause of mortality in modern societies; therefore, improved and more reliablein vitrocancer models are needed to expedite fundamental research and anti-cancer drug development. Here, we describe the use of a miniaturized continuous stirred tank reactor (mCSTR) to first fabricate and mature cancer spheroids (i.e. derived from MCF7 cells, DU145 cells, and a mix of MCF7 cells and fibroblasts), and then to conduct anti-cancer drug assays under continuous perfusion. This 3 ml mCSTR features an off-center agitation system that enables homogeneous chaotic laminar mixing at low speeds to support cell aggregation. We incubated cell suspensions for 3 d in ultra-low-attachment plates to allow formation of discoid cell aggregates (∼600µm in diameter). These cell aggregates were then transferred into mCSTRs and continuously fed with culture medium. We characterized the spheroid morphology and the expression of relevant tumor biomarkers at different maturation times for up to 4 weeks. The spheroids progressively increased in size during the first 5-6 d of culture to reach a steady diameter between 600 and 800µm. In proof-of-principle experiments, we demonstrated the use of this mCSTR in anti-cancer drug testing. Three drugs commonly used in breast cancer treatment (doxorubicin, docetaxel, and paclitaxel) were probed at different concentrations in MCF7-derived spheroids. In these experiments, we evaluated cell viability, glucose consumption, spheroid morphology, lactate dehydrogenase activity, and the expression of genes associated with drug resistance (ABCB1andABCC1) and anti-apoptosis (Bcl2). We envision the use of this agitated system as a tumor-on-a-chip platform to expedite efficacy and safety testing of novel anti-cancer drugs and possibly in personalized medicine applications.

The Tumor-on-Chip: Recent Advances in the Development of Microfluidic Systems to Recapitulate the Physiology of Solid Tumors.

The ideal in vitro recreation of the micro-tumor niche-although much needed for a better understanding of cancer etiology and development of better anticancer therapies-is highly challenging. Tumors are complex three-dimensional (3D) tissues that establish a dynamic cross-talk with the surrounding tissues through complex chemical signaling. An extensive body of experimental evidence has established that 3D culture systems more closely recapitulate the architecture and the physiology of human solid tumors when compared with traditional 2D systems. Moreover, conventional 3D culture systems fail to recreate the dynamics of the tumor niche. Tumor-on-chip systems, which are microfluidic devices that aim to recreate relevant features of the tumor physiology, have recently emerged as powerful tools in cancer research. In tumor-on-chip systems, the use of microfluidics adds another dimension of physiological mimicry by allowing a continuous feed of nutrients (and pharmaceutical compounds). Here, we discuss recently published literature related to the culture of solid tumor-like tissues in microfluidic systems (tumor-on-chip devices). Our aim is to provide the readers with an overview of the state of the art on this particular theme and to illustrate the toolbox available today for engineering tumor-like structures (and their environments) in microfluidic devices. The suitability of tumor-on-chip devices is increasing in many areas of cancer research, including the study of the physiology of solid tumors, the screening of novel anticancer pharmaceutical compounds before resourcing to animal models, and the development of personalized treatments. In the years to come, additive manufacturing (3D bioprinting and 3D printing), computational fluid dynamics, and medium- to high-throughput omics will become powerful enablers of a new wave of more sophisticated and effective tumor-on-chip devices.

SARS: mobilizing and maintaining a public health emergency response.

During the spring and summer of 2003, the Centers for Disease Control and Prevention (CDC) mobilized the resources of the entire agency in a concerted effort to meet the challenges posed by the outbreak of Severe Acute Respiratory Syndrome (SARS). Over the 133 days that comprised the emergency response phase of the SARS outbreak, CDC utilized the skills of more than 850 people. These staff were deployed from every Center, Institute, and Office within CDC and the Agency for Toxic Substances and Disease Registry. They provided technical assistance to countries reporting large numbers of cases and requesting assistance, met passengers and crew from these locations upon arrival in the United States, and assured that the syndrome was reported and thoroughly investigated within the United States. This paper describes the operational requirements that were established and the resources that were used to conduct this investigation.