Evidence for Two Subpopulations of Cerebrospinal Fluid-Contacting Neurons with Opposite GABAergic Signaling in Adult Mouse Spinal Cord.

Spinal cerebrospinal fluid-contacting neurons (CSF-cNs) form an evolutionary conserved bipolar cell population localized around the central canal of all vertebrates. CSF-cNs were shown to express molecular markers of neuronal immaturity into adulthood; however, the impact of their incomplete maturation on the chloride (Cl-) homeostasis as well as GABAergic signaling remains unknown. Using adult mice from both sexes, in situ hybridization revealed that a proportion of spinal CSF-cNs (18.3%) express the Na+-K+-Cl- cotransporter 1 (NKCC1) allowing intracellular Cl- accumulation. However, we did not find expression of the K+-Cl- cotransporter 2 (KCC2) responsible for Cl- efflux in any CSF-cNs. The lack of KCC2 expression results in low Cl- extrusion capacity in CSF-cNs under high Cl- load in whole-cell patch clamp. Using cell-attached patch clamp allowing recordings with intact intracellular Cl- concentration, we found that the activation of ionotropic GABAA receptors (GABAA-Rs) induced both depolarizing and hyperpolarizing responses in CSF-cNs. Moreover, depolarizing GABA responses can drive action potentials as well as intracellular calcium elevations by activating voltage-gated calcium channels. Blocking NKCC1 with bumetanide inhibited the GABA-induced calcium transients in CSF-cNs. Finally, we show that metabotropic GABAB receptors have no hyperpolarizing action on spinal CSF-cNs as their activation with baclofen did not mediate outward K+ currents, presumably due to the lack of expression of G-protein-coupled inwardly rectifying potassium (GIRK) channels. Together, these findings outline subpopulations of spinal CSF-cNs expressing inhibitory or excitatory GABAA-R signaling. Excitatory GABA may promote the maturation and integration of young CSF-cNs into the existing spinal circuit.

Maternal Pre-Existing Diabetes: A Non-Inherited Risk Factor for Congenital Cardiopathies.

Congenital heart defects (CHDs) are the most common form of birth defects in humans. They occur in 9 out of 1000 live births and are defined as structural abnormalities of the heart. Understanding CHDs is difficult due to the heterogeneity of the disease and its multifactorial etiology. Advances in genomic sequencing have made it possible to identify the genetic factors involved in CHDs. However, genetic origins have only been found in a minority of CHD cases, suggesting the contribution of non-inherited (environmental) risk factors to the etiology of CHDs. Maternal pregestational diabetes is associated with a three- to five-fold increased risk of congenital cardiopathies, but the underlying molecular mechanisms are incompletely understood. According to current hypotheses, hyperglycemia is the main teratogenic agent in diabetic pregnancies. It is thought to induce cell damage, directly through genetic and epigenetic dysregulations and/or indirectly through production of reactive oxygen species (ROS). The purpose of this review is to summarize key findings on the molecular mechanisms altered in cardiac development during exposure to hyperglycemic conditions in utero. It also presents the various in vivo and in vitro techniques used to experimentally model pregestational diabetes. Finally, new approaches are suggested to broaden our understanding of the subject and develop new prevention strategies.

Decoding loneliness: Can explainable AI help in understanding language differences in lonely older adults?

STUDY OBJECTIVES: Loneliness impacts the health of many older adults, yet effective and targeted interventions are lacking. Compared to surveys, speech data can capture the personalized experience of loneliness. In this proof-of-concept study, we used Natural Language Processing to extract novel linguistic features and AI approaches to identify linguistic features that distinguish lonely adults from non-lonely adults. METHODS: Participants completed UCLA loneliness scales and semi-structured interviews (sections: social relationships, loneliness, successful aging, meaning/purpose in life, wisdom, technology and successful aging). We used the Linguistic Inquiry and Word Count (LIWC-22) program to analyze linguistic features and built a classifier to predict loneliness. Each interview section was analyzed using an explainable AI (XAI) model to classify loneliness. RESULTS: The sample included 97 older adults (age 66-101 years, 65 % women). The model had high accuracy (Accuracy: 0.889, AUC: 0.8), precision (F1: 0.8), and recall (1.0). The sections on social relationships and loneliness were most important for classifying loneliness. Social themes, conversational fillers, and pronoun usage were important features for classifying loneliness. CONCLUSIONS: XAI approaches can be used to detect loneliness through the analyses of unstructured speech and to better understand the experience of loneliness.

Understanding the cell fate and behavior of progenitors at the origin of the mouse cardiac mitral valve.

Congenital heart malformations include mitral valve defects, which remain largely unexplained. During embryogenesis, a restricted population of endocardial cells within the atrioventricular canal undergoes an endothelial-to-mesenchymal transition to give rise to mitral valvular cells. However, the identity and fate decisions of these progenitors as well as the behavior and distribution of their derivatives in valve leaflets remain unknown. We used single-cell RNA sequencing (scRNA-seq) of genetically labeled endocardial cells and microdissected mouse embryonic and postnatal mitral valves to characterize the developmental road. We defined the metabolic processes underlying the specification of the progenitors and their contributions to subtypes of valvular cells. Using retrospective multicolor clonal analysis, we describe specific modes of growth and behavior of endocardial cell-derived clones, which build up, in a proper manner, functional valve leaflets. Our data identify how both genetic and metabolic mechanisms specifically drive the fate of a subset of endocardial cells toward their distinct clonal contribution to the formation of the valve.

Nuclei Isolation from Mouse Cardiac Progenitor Cells for Epigenome and Gene Expression Profiling at Single-Cell Resolution.

The developing heart is a complex structure containing various progenitor cells controlled by complex regulatory mechanisms. The examination of the gene expression and chromatin state of individual cells allows the identification of the cell type and state. Single-cell sequencing approaches have revealed a number of important characteristics of cardiac progenitor cell heterogeneity. However, these methods are generally restricted to fresh tissue, which limits studies with diverse experimental conditions, as the fresh tissue must be processed at once in the same run to reduce the technical variability. Therefore, easy and flexible procedures to produce data from methods such as single-nucleus RNA sequencing (snRNA-seq) and the single-nucleus assay for transposase-accessible chromatin with high-throughput sequencing (snATAC-seq) are needed in this area. Here, we present a protocol to rapidly isolate nuclei for subsequent single-nuclei dual-omics (combined snRNA-seq and snATAC-seq). This method allows the isolation of nuclei from frozen samples of cardiac progenitor cells and can be combined with platforms that use microfluidic chambers.

"Say Ahhh": Experience and Views on Halitosis Management in the General Public in Victoria, Australia.

INTRODUCTION: Halitosis is common and can have a significant impact on quality of life. It is attributed to both intraoral and extraoral causes. Although halitosis treatment depends on the aetiology, little is known about consumers' perception of halitosis causes and the types of assistance sought. The Say Ahhh Study aimed to explore the interprofessional care for halitosis and improve understanding by the general public. Phase 1 explored the perceptions of health care providers. This article reports the findings of Phase 2, which explored the general public's knowledge of halitosis, its management, and help-seeking behaviour. METHODS: Patients and clients at 2 medical clinics, 3 dental clinics, and 2 pharmacies in Melbourne and rural Victoria were approached and invited to participate in a short semistructured interview. Qualitative data was thematically analysed. RESULTS: A total of 122 participants (54 males, 66 females, 2 missing) were interviewed. Participants' past experience with halitosis influences their understanding of the cause. Halitosis was attributed mostly to poor oral hygiene and diet and less commonly to systemic disease. Their perception of the condition's severity influenced whether they seek professional help. Their perceived roles of health professionals and ease of access influenced their choice of health professionals to seek help from. CONCLUSION: The Victorian general public is aware of the causes of halitosis and the available treatment options. Choice of treatment and help-seeking behaviour are influenced by experience, perceived severity of the condition, and perceived role of health professionals and their accessibility.