Proper acquisition of cell class identity in organoids allows definition of fate specification programs of the human cerebral cortex.

Realizing the full utility of brain organoids to study human development requires understanding whether organoids precisely replicate endogenous cellular and molecular events, particularly since acquisition of cell identity in organoids can be impaired by abnormal metabolic states. We present a comprehensive single-cell transcriptomic, epigenetic, and spatial atlas of human cortical organoid development, comprising over 610,000 cells, from generation of neural progenitors through production of differentiated neuronal and glial subtypes. We show that processes of cellular diversification correlate closely to endogenous ones, irrespective of metabolic state, empowering the use of this atlas to study human fate specification. We define longitudinal molecular trajectories of cortical cell types during organoid development, identify genes with predicted human-specific roles in lineage establishment, and uncover early transcriptional diversity of human callosal neurons. The findings validate this comprehensive atlas of human corticogenesis in vitro as a resource to prime investigation into the mechanisms of human cortical development.

3D Brain Organoids: Studying Brain Development and Disease Outside the Embryo.

Scientists have been fascinated by the human brain for centuries, yet knowledge of the cellular and molecular events that build the human brain during embryogenesis and of how abnormalities in this process lead to neurological disease remains very superficial. In particular, the lack of experimental models for a process that largely occurs during human in utero development, and is therefore poorly accessible for study, has hindered progress in mechanistic understanding. Advances in stem cell-derived models of human organogenesis, in the form of three-dimensional organoid cultures, and transformative new analytic technologies have opened new experimental pathways for investigation of aspects of development, evolution, and pathology of the human brain. Here, we consider the biology of brain organoids, compared and contrasted with the endogenous human brain, and highlight experimental strategies to use organoids to pioneer new understanding of human brain pathology.

Autism genes converge on asynchronous development of shared neuron classes.

Genetic risk for autism spectrum disorder (ASD) is associated with hundreds of genes spanning a wide range of biological functions1-6. The alterations in the human brain resulting from mutations in these genes remain unclear. Furthermore, their phenotypic manifestation varies across individuals7,8. Here we used organoid models of the human cerebral cortex to identify cell-type-specific developmental abnormalities that result from haploinsufficiency in three ASD risk genes-SUV420H1 (also known as KMT5B), ARID1B and CHD8-in multiple cell lines from different donors, using single-cell RNA-sequencing (scRNA-seq) analysis of more than 745,000 cells and proteomic analysis of individual organoids, to identify phenotypic convergence. Each of the three mutations confers asynchronous development of two main cortical neuronal lineages-γ-aminobutyric-acid-releasing (GABAergic) neurons and deep-layer excitatory projection neurons-but acts through largely distinct molecular pathways. Although these phenotypes are consistent across cell lines, their expressivity is influenced by the individual genomic context, in a manner that is dependent on both the risk gene and the developmental defect. Calcium imaging in intact organoids shows that these early-stage developmental changes are followed by abnormal circuit activity. This research uncovers cell-type-specific neurodevelopmental abnormalities that are shared across ASD risk genes and are finely modulated by human genomic context, finding convergence in the neurobiological basis of how different risk genes contribute to ASD pathology.

Cell-type specific defects in PTEN-mutant cortical organoids converge on abnormal circuit activity.

De novo heterozygous loss-of-function mutations in phosphatase and tensin homolog (PTEN) are strongly associated with autism spectrum disorders; however, it is unclear how heterozygous mutations in this gene affect different cell types during human brain development and how these effects vary across individuals. Here, we used human cortical organoids from different donors to identify cell-type specific developmental events that are affected by heterozygous mutations in PTEN. We profiled individual organoids by single-cell RNA-seq, proteomics and spatial transcriptomics and revealed abnormalities in developmental timing in human outer radial glia progenitors and deep-layer cortical projection neurons, which varied with the donor genetic background. Calcium imaging in intact organoids showed that both accelerated and delayed neuronal development phenotypes resulted in similar abnormal activity of local circuits, irrespective of genetic background. The work reveals donor-dependent, cell-type specific developmental phenotypes of PTEN heterozygosity that later converge on disrupted neuronal activity.

Individual brain organoids reproducibly form cell diversity of the human cerebral cortex.

Experimental models of the human brain are needed for basic understanding of its development and disease1. Human brain organoids hold unprecedented promise for this purpose; however, they are plagued by high organoid-to-organoid variability2,3. This has raised doubts as to whether developmental processes of the human brain can occur outside the context of embryogenesis with a degree of reproducibility that is comparable to the endogenous tissue. Here we show that an organoid model of the dorsal forebrain can reliably generate a rich diversity of cell types appropriate for the human cerebral cortex. We performed single-cell RNA-sequencing analysis of 166,242 cells isolated from 21 individual organoids, finding that 95% of the organoids generate a virtually indistinguishable compendium of cell types, following similar developmental trajectories and with a degree of organoid-to-organoid variability comparable to that of individual endogenous brains. Furthermore, organoids derived from different stem cell lines show consistent reproducibility in the cell types produced. The data demonstrate that reproducible development of the complex cellular diversity of the central nervous system does not require the context of the embryo, and that establishment of terminal cell identity is a highly constrained process that can emerge from diverse stem cell origins and growth environments.

Effects of different types of leisure activities on working memory across the adult lifespan.

The aim of the present study was to contrast the potential influence of five independent types of leisure activities (physical, mental, social, cultural, and passive) on working memory in a lifespan sample and in specific stages of adulthood (young, middle-aged, and older adults). A sample of 1652 healthy adults between 21 and 80 years of age participated in the study. Leisure activities were assessed through a lifestyle questionnaire created for the study. Working memory was measured in the verbal and spatial domains using a computerized n-back task that allowed us to reliably measure discrimination and reaction times. Across adulthood, mental (computer use and hobbies) and social leisure activities predicted greater verbal and spatial working memory discrimination; mental (reading) and social activities predicted faster verbal working memory; and mental (computer use) and physical activities predicted faster spatial working memory. In young adults, mental (computer use) and social activities were associated with greater verbal and spatial working memory performance. In middle-aged adults, physical and mental activities (computer use) were associated with greater working memory performance. In older adults, physical, mental (hobbies), and social activities were associated with greater working memory performance. Leisure activities can enhance working memory discrimination and speed independent of individuals' age.

Nutrient effects on working memory across the adult lifespan.

OBJECTIVE: To identify the nutrients that influence the performance of working memory, which is greatly affected as age progresses. METHOD: A total of 1646 healthy adults between 21 and 80 years old participated in the study. The daily consumption of 64 nutrients was examined using a food frequency questionnaire that assessed food intake during the previous year. Working memory was measured in the verbal and spatial domains using a computerized task. We examined which nutrients influence working memory across the entire adult lifespan and whether the influence of any of these nutrients on working memory is moderated by individuals' ages. RESULTS: Working memory, across the entire adult lifespan, benefits from the intake of cholesterol, alcohol, gamma- and delta-tocopherol, vitamin B6, and palmitoleic, oleic, alpha linoleic and linoleic acids. Moderator analyses revealed that fats, energy, lactose and sodium negatively influenced working memory in middle-aged and older adults, whereas vitamin D and vitamin C had positive effects on memory beyond 70 years of age. CONCLUSION: Nutrients have the ability to positively or negatively affect working memory, which varies as a function of age.

Protein carbonylation in food and nutrition: a concise update.

Protein oxidation is a topic of indisputable scientific interest given the impact of oxidized proteins on food quality and safety. Carbonylation is regarded as one of the most notable post-translational modifications in proteins and yet, this reaction and its consequences are poorly understood. From a mechanistic perspective, primary protein carbonyls (i.e. α-aminoadipic and γ-glutamic semialdehydes) have been linked to radical-mediated oxidative stress, but recent studies emphasize the role alternative carbonylation pathways linked to the Maillard reaction. Secondary protein carbonyls are introduced in proteins via covalent linkage of lipid carbonyls (i.e. protein-bound malondialdehyde). The high reactivity of protein carbonyls in foods and other biological systems indicates the intricate chemistry of these species and urges further research to provide insight into these molecular mechanisms and pathways. In particular, protein carbonyls are involved in the formation of aberrant and dysfunctional protein aggregates, undergo further oxidation to yield carboxylic acids of biological relevance and establish interactions with other biomolecules such as oxidizing lipids and phytochemicals. From a methodological perspective, the routine dinitrophenylhydrazine (DNPH) method is criticized not only for the lack of accuracy and consistency but also authors typically perform a poor interpretation of DNPH results, which leads to misleading conclusions. From a practical perspective, the biological relevance of protein carbonyls in the field of food science and nutrition is still a topic of debate. Though the implication of carbonylation on impaired protein functionality and poor protein digestibility is generally recognized, the underlying mechanism of such connections requires further clarification. From a medical perspective, protein carbonyls are highlighted as markers of protein oxidation, oxidative stress and disease. Yet, the specific role of specific protein carbonyls in the onset of particular biological impairments needs further investigations. Recent studies indicates that regardless of the origin (in vivo or dietary) protein carbonyls may act as signalling molecules which activate not only the endogenous antioxidant defences but also implicate the immune system. The present paper concisely reviews the most recent advances in this topic to identify, when applicable, potential fields of interest for future studies.

Effects of hypotension and hypertension on source memory and working memory.

OBJECTIVES: The effects of chronic low and high blood pressure on memory are unclear due to divergent results, originating in part due to participant misclassifications. The aim of this study was to compare source memory and working memory performance in individuals diagnosed with hypotension or hypertension with the performance of normotensive participants. Hypertensive and hypotensive individuals were receiving medical treatment. METHOD: From a sample of 1656 participants, 219 were identified as hypertensive, and 37 were identified as hypotensive. Each of these two groups was compared with normotensive individuals matched by age, education and sex. Source memory performance and working memory performance were assessed through computerized tasks. RESULTS: Source memory accuracy was poorer in hypotensive and hypertensive individuals than in normotensive individuals, and spatial working memory discrimination was inferior in hypertensive participants compared to normotensive individuals. CONCLUSION: Blood pressure impairment should be considered a major concern because it has been linked to severe cardiovascular and cerebrovascular diseases. Furthermore, here we show that it has negative effects on the two types of memory that are most essential for preserving a self-sufficient lifestyle.

Metamemory Mediates the Effects of Age on Episodic and Working Memory across the Adult Lifespan.

BACKGROUND: Previous studies have examined the direct relationship between metamemory and memory performance in young and older adults, but the results of these studies have been inconsistent. Therefore, we examined whether metamemory mediates the effects of age on memory performance. METHODS: We examined episodic memory and working memory through computerized tasks performed by a lifespan sample of 1554 healthy adults. Seven metamemory traits were measured with the Metamemory in Adulthood (MIA) questionnaire. Separate structural equation modeling analyses were conducted to investigate potential metamemory mediators that intervened between age and the accuracy and speed of accessing information from episodic and working memory. RESULTS: The use of internal or external strategies mediated the effects of age on episodic memory and spatial working memory performance. The perception of one's own memory capacity and the experience of anxiety when using memory functions mediated the effects of age on working memory performance in both domains. CONCLUSIONS: Metamemory traits have the power to strengthen or weaken the course of episodic and working memory decline throughout adulthood.

Contributions of Cognitive Aging Models to the Explanation of Source Memory Decline across the Adult Lifespan.

Background: A number of cognitive aging models have been proposed to explain the age-related decline in several cognitive functions, but these models have rarely been examined together. We analyzed the contributions of four main models - processing resources, speed of processing, cognitive reserve and knowledge - to source memory decay related to the aging process.Methods: A total of 1554 healthy adults between 21 and 80 years old participated in the study. Structural equation modeling was conducted on data from the whole sample and separately in the data from young, middle-aged and older adult age groups. To estimate each cognitive model, we measured working memory discrimination levels (processing resources), working memory reaction times (speed of processing), education (cognitive reserve) and vocabulary (knowledge).Results: Processing resources mediate the effects of age on source memory across the adult lifespan, whereas speed of processing mediates these effects only in young adults, cognitive reserve only in middle-aged adults and knowledge only in older adults.Conclusions: Processing resources was the cognitive model that most contributes to explaining source memory decay. The fact that the other models are relevant to specific age groups provides useful information to exploit their benefits to preserve source memory in specific life stages.

Multidimensional Patterns of Sexual Risk Behavior and Psychiatric Disorders in Men with Substance Use Disorders.

Previous evidence links substance use disorders (SUD) to STI/HIV risk and suggests that comorbid psychiatric disorders increase the probability to engage in sexual risk behaviors. This study had two aims: (1) to identify subgroups based on sexual risk behavior using a person-centered approach in a sample of substance users and (2) to measure the association of psychiatric and SUD with subgroup membership. We assessed 402 male adults with SUD, reporting sexual intercourse in the previous 12 months using the HIV-Risk Behavior Scale and the Mini International Neuropsychiatric Interview. Latent class analysis was performed to determine multidimensional patterns of sexual risk behaviors and multinomial logistic regression was utilized to associate classes with disorders. The three-class model showed the best fit, and the classes were labeled: Relationship-Based (31.34% of the sample), Condom-Based (39.55%), and Multiple Risks (29.10%). Controlling for age and marital status, major depressive disorders, antisocial personality disorder, and any psychiatric disorder were associated with the Multiple Risks class. Results stress the importance of developing a personalized assessment and counseling for sexual risk behaviors in individuals with SUD, particularly when they endorse criteria for comorbid psychiatric disorders. Future studies should focus on evaluating differential response to preventive interventions.

Deconvolving sequence features that discriminate between overlapping regulatory annotations.

Genomic loci with regulatory potential can be annotated with various properties. For example, genomic sites bound by a given transcription factor (TF) can be divided according to whether they are proximal or distal to known promoters. Sites can be further labeled according to the cell types and conditions in which they are active. Given such a collection of labeled sites, it is natural to ask what sequence features are associated with each annotation label. However, discovering such label-specific sequence features is often confounded by overlaps between the labels; e.g. if regulatory sites specific to a given cell type are also more likely to be promoter-proximal, it is difficult to assess whether motifs identified in that set of sites are associated with the cell type or associated with promoters. In order to meet this challenge, we developed SeqUnwinder, a principled approach to deconvolving interpretable discriminative sequence features associated with overlapping annotation labels. We demonstrate the novel analysis abilities of SeqUnwinder using three examples. Firstly, SeqUnwinder is able to unravel sequence features associated with the dynamic binding behavior of TFs during motor neuron programming from features associated with chromatin state in the initial embryonic stem cells. Secondly, we characterize distinct sequence properties of multi-condition and cell-specific TF binding sites after controlling for uneven associations with promoter proximity. Finally, we demonstrate the scalability of SeqUnwinder to discover cell-specific sequence features from over one hundred thousand genomic loci that display DNase I hypersensitivity in one or more ENCODE cell lines.

Prediction of Cesarean Section for Intrapartum Fetal Compromise: A Multivariable Model from a Prospective Observational Approach.

OBJECTIVE: A cesarean section for intrapartum fetal compromise (IFC) is performed to avoid potential damage to the newborn. It is, therefore, crucial to develop an accurate prediction model that can anticipate, prior to labor, which fetus may be at risk of presenting this condition. MATERIAL AND METHODS: To calculate a prediction model for IFC, the clinical, epidemiological, and ultrasonographic variables of 538 patients admitted to the maternity of La Fe Hospital were studied and evaluated using univariable and multivariable logistic regression analysis, using the area under the curve (AUC) and the Akaike Information Criteria (AIC). RESULTS: In the univariable analysis, CPR MoM was the best single parameter for the prediction of CS for IFC (OR 0.043, p < 0.0001; AUC 0.72, p < 0.0001). Concerning the multivariable analysis, for the general population, the best prediction model (lower AIC) included the CPR multiples of the median (MoM), the maternal age, height, and parity, the smoking habits, and the type of labor onset (spontaneous or induction) (AUC 0.80, p < 0.0001). In contrast, for the pregnancies undergoing labor induction, the best prediction model included the CPR MoM, the maternal height and parity, and the smoking habits (AUC 0.80, p < 0.0001). None of the models included estimated fetal weight (EFW). CONCLUSIONS: CS for IFC can be moderately predicted prior to labor using maternal characteristics and CPR MoM. A validation study is pending to apply these models in daily clinical practice.

Prediction of Failure to Progress after Labor Induction: A Multivariable Model Using Pelvic Ultrasound and Clinical Data.

Objective: Labor induction is one of the leading causes of obstetric admission. This study aimed to create a simple model for predicting failure to progress after labor induction using pelvic ultrasound and clinical data. Material and Methods: A group of 387 singleton pregnant women at term with unruptured amniotic membranes admitted for labor induction were included in an observational prospective study. Clinical and ultrasonographic variables were collected at admission prior to the onset of contractions, and labor data were collected after delivery. Multivariable logistic regression analysis was applied to create several models to predict cesarean section due to failure to progress. Afterward, the most accurate and reproducible model was selected according to the lowest Akaike Information Criteria (AIC) with a high area under the curve (AUC). Results: Plausible parameters for explaining failure to progress were initially obtained from univariable analysis. With them, several multivariable analyses were evaluated. Those parameters with the highest reproducibility included maternal age (p < 0.05), parity (p < 0.0001), fetal gender (p < 0.05), EFW centile (p < 0.01), cervical length (p < 0.01), and posterior occiput position (p < 0.001), but the angle of descent was disregarded. This model obtained an AIC of 318.3 and an AUC of 0.81 (95% CI 0.76-0.86, p < 0.0001) with detection rates of 24% and 37% for FPRs of 5% and 10%. Conclusions: A simplified clinical and sonographic model may guide the management of pregnancies undergoing labor induction, favoring individualized patient management.

Cerebroplacental Ratio as a Predictive Factor of Emergency Cesarean Sections for Intrapartum Fetal Compromise: A Systematic Review.

Background: This systematic review aimed to clarify the association between the cerebroplacental ratio (CPR) and emergency cesarean sections (CSs) due to intrapartum fetal compromise (IFC). Methods: Datasets of PubMed, ScienceDirect, CENTRAL, Embase, and Google Scholar were searched for studies published up to January 2024 regarding the relationship between the CPR and the rate of CS for IFC, as well as the predictive value of the CPR. Results: The search identified 582 articles, of which 16 observational studies were finally included, most of them with a prospective design. A total of 14,823 patients were involved. A low CPR was associated with a higher risk of CS for IFC. The predictive value of the CPR was very different among the studies due to substantial heterogeneity regarding the group of patients included and the time interval from CPR evaluation to delivery. Conclusions: A low CPR is associated with a higher risk of CS for IFC, although with a poor predictive value. The CPR could be calculated prior to labor in all patients to stratify the risk of CS due to IFC.

Neuronal Galectin-4 is required for axon growth and for the organization of axonal membrane L1 delivery and clustering.

Axon membrane glycoproteins are essential for neuronal differentiation, although the mechanisms underlying their polarized sorting and organization are poorly understood. We describe here that galectin-4 (Gal-4), a lectin highly expressed in gastrointestinal tissues and involved in epithelial glycoprotein transport, is expressed by hippocampal and cortical neurons where it is sorted to discrete segments of the axonal membrane in a microtubule- and sulfatide-dependent manner. Gal-4 knockdown retards axon growth, an effect that can be rescued by recombinant Gal-4 addition. This Gal-4 reduction, as inhibition of sulfatide synthesis does, lowers the presence and clustered organization of axon growth-promoting molecule NCAM L1 at the axon membrane. Furthermore, we find that Gal-4 interacts with L1 by specifically binding to LacNAc branch ends of L1 N-glycans. Impairing the maturation of these N-glycans precludes Gal-4/L1 association resulting in a failure of L1 membrane cluster organization. In all, Gal-4 sorts to axon plasma membrane segments by binding to sulfatide-containing microtubule-associated carriers and being bivalent, it organizes the transport of L1, and likely other axonal glycoproteins, by attaching them to the carriers through their LacNAc termini. This mechanism would underlie L1 functional organization on the plasma membrane, required for proper axon growth.

ERK1/2 activation in heart is controlled by melusin, focal adhesion kinase and the scaffold protein IQGAP1.

Extracellular signal-regulated kinase 1/2 (ERK1/2) signalling is a key pathway in cardiomyocyte hypertrophy and survival in response to many different stress stimuli. We have previously characterized melusin as a muscle-specific chaperone protein capable of ERK1/2 signalling activation in the heart. Here, we show that in the heart, melusin forms a supramolecular complex with the proto-oncogene c-Raf, MEK1/2 (also known as MAPKK1/2) and ERK1/2 and that melusin-bound mitogen-activated protein kinases (MAPKs) are activated by pressure overload. Moreover, we demonstrate that both focal adhesion kinase (FAK) and IQ motif-containing GTPase activating protein 1 (IQGAP1), a scaffold protein for the ERK1/2 signalling cascade, are part of the melusin complex and are required for ERK1/2 activation in response to pressure overload. Finally, analysis of isolated neonatal cardiomyocytes indicates that both FAK and IQGAP1 regulate melusin-dependent cardiomyocyte hypertrophy and survival through ERK1/2 activation.

Intestinal permeability assessment using lactulose and mannitol in celiac disease.

Alterations in intestinal permeability can lead to increased uptake of luminal antigens, which has been linked to several intestinal diseases, such as inflammatory bowel diseases, celiac disease, and irritable bowel syndrome, but also to extra-intestinal diseases. Promising therapies that target intestinal permeability could be developed, for instance tight junction modulators. Consequently, permeability assays are increasingly being used as treatment endpoints in clinical studies. Therefore, reliable, reproducible, and feasible methods for measuring intestinal permeability in the clinical setting are necessary. Currently, a variety of in vivo, ex vivo, and in vitro tests are available, some of which are only applicable to basic research. Despite the various options available to measure gut permeability, their use in clinical setting is still limited because of their heterogeneity. Here, we describe a clinical method to measure intestinal permeability using two non-metabolizable sugars.