A genome-wide arrayed CRISPR screen identifies PLSCR1 as an intrinsic barrier to SARS-CoV-2 entry that recent virus variants have evolved to resist.

Interferons (IFNs) play a crucial role in the regulation and evolution of host-virus interactions. Here, we conducted a genome-wide arrayed CRISPR knockout screen in the presence and absence of IFN to identify human genes that influence Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection. We then performed an integrated analysis of genes interacting with SARS-CoV-2, drawing from a selection of 67 large-scale studies, including our own. We identified 28 genes of high relevance in both human genetic studies of Coronavirus Disease 2019 (COVID-19) patients and functional genetic screens in cell culture, with many related to the IFN pathway. Among these was the IFN-stimulated gene PLSCR1. PLSCR1 did not require IFN induction to restrict SARS-CoV-2 and did not contribute to IFN signaling. Instead, PLSCR1 specifically restricted spike-mediated SARS-CoV-2 entry. The PLSCR1-mediated restriction was alleviated by TMPRSS2 overexpression, suggesting that PLSCR1 primarily restricts the endocytic entry route. In addition, recent SARS-CoV-2 variants have adapted to circumvent the PLSCR1 barrier via currently undetermined mechanisms. Finally, we investigate the functional effects of PLSCR1 variants present in humans and discuss an association between PLSCR1 and severe COVID-19 reported recently.

TGF-ß and RAS jointly unmask primed enhancers to drive metastasis.

Epithelial-to-mesenchymal transitions (EMTs) and extracellular matrix (ECM) remodeling are distinct yet important processes during carcinoma invasion and metastasis. Transforming growth factor ß (TGF-ß) and RAS, signaling through SMAD and RAS-responsive element-binding protein 1 (RREB1), jointly trigger expression of EMT and fibrogenic factors as two discrete arms of a common transcriptional response in carcinoma cells. Here, we demonstrate that both arms come together to form a program for lung adenocarcinoma metastasis and identify chromatin determinants tying the expression of the constituent genes to TGF-ß and RAS inputs. RREB1 localizes to H4K16acK20ac marks in histone H2A.Z-loaded nucleosomes at enhancers in the fibrogenic genes interleukin-11 (IL11), platelet-derived growth factor-B (PDGFB), and hyaluronan synthase 2 (HAS2), as well as the EMT transcription factor SNAI1, priming these enhancers for activation by a SMAD4-INO80 nucleosome remodeling complex in response to TGF-ß. These regulatory properties segregate the fibrogenic EMT program from RAS-independent TGF-ß gene responses and illuminate the operation and vulnerabilities of a bifunctional program that promotes metastatic outgrowth.

Fibrinolysis associated proteins and lipopolysaccharide bioactivity in plasma and cerebrospinal fluid in multiple sclerosis.

The coagulation cascade and fibrinolysis have links with neuroinflammation and increased activation of the coagulation system has been reported in MS patients. We quantified levels of D-dimer, tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1) and the bioactivity of bacterial lipopolysaccharide (LPS) in cerebrospinal fluid (CSF) and plasma from newly diagnosed untreated MS patients and controls. These molecules showed multiple correlations with each other as well as with age, HLA-DRB1*15:01, body-mass-index and CSF IgG. Our results confirm previous findings of increased plasma PAI-1 and LPS in MS patients compared to controls indicating changes in platelet function and gut permeability in MS.

From asylums to deinstitutionalization and after: an analytic review.

BACKGROUND: After a period of institution-based mental health care, in which the asylum system was the way in which the mental patients were treated, deinstitutionalization brought a set of significant changes and transformations in the conceptualization of mental illness and, by extension, the traditional therapeutic settings in which those in most need were assisted. However, this shift in the psychiatric domain was not only accompanied by valued achievements, but also by difficulties and challenges, as has been evidenced today. AIM/OBJECTIVE: The aim of this paper is thus to examine the pros and cons of the closure of asylums, and the subsequent implementation of deinstitutionalization over the 60 years or so of such important transformations in the field of psychiatry. METHODS: In considering this question, I examine in detail recent works of literature based on scholarly knowledge. In addition, I identify various issues involved, as well as ways of confronting these so as to attempt to overcome the difficulties they present. RESULTS AND CONCLUSIONS: As I show here, the changes in the treatment and care of the mentally ill after asylum and deinstitutionalization brought a new air of hope to patients and their families, but also had undesirable effects. The paper also considers how mental health professionals today try to solve these effects on behalf of patients and society as a whole.

Circulating platelets modulate oligodendrocyte progenitor cell differentiation during remyelination.

Revealing unknown cues that regulate oligodendrocyte progenitor cell (OPC) function in remyelination is important to optimise the development of regenerative therapies for multiple sclerosis (MS). Platelets are present in chronic non-remyelinated lesions of MS and an increase in circulating platelets has been described in experimental autoimmune encephalomyelitis (EAE) mice, an animal model for MS. However, the contribution of platelets to remyelination remains unexplored. Here we show platelet aggregation in proximity to OPCs in areas of experimental demyelination. Partial depletion of circulating platelets impaired OPC differentiation and remyelination, without altering blood-brain barrier stability and neuroinflammation. Transient exposure to platelets enhanced OPC differentiation in vitro, whereas sustained exposure suppressed this effect. In a mouse model of thrombocytosis (Calr+/-), there was a sustained increase in platelet aggregation together with a reduction of newly-generated oligodendrocytes following toxin-induced demyelination. These findings reveal a complex bimodal contribution of platelet to remyelination and provide insights into remyelination failure in MS.

Lack of autonomy and professional recognition as major factors for burnout in midwives: A systematic mixed-method review.

AIM: The aim of this study was to review the existing evidence on burnout levels in midwives and the main related factors. DESIGN: Mixed studies systematic review. DATA SOURCES: PubMed, Scopus and Web of Science were sourced from 2018 and 2023. REVIEW METHODS: Inclusion criteria: quantitative cross-sectional or qualitative articles published in English within the last 5 years. EXCLUSION CRITERIA: studies with undergraduate or trainee midwives, studies examining the factors in a pandemic setting and those not answering the research question. Potential risk of bias was assessed using the Mixed Methods Assessment Tool (MMAT). A convergent synthesis design was followed through a thematic synthesis using Thomas and Harden's three-step method: inductive coding of the text, development of descriptive themes and generation of analytical themes. Qualitative approaches adopted exploratory descriptive studies and participatory action research. RESULTS: Thirty-six studies were included, with a total of 17,364 participants. There were higher levels of burnout in midwives who were single, under 35-40 years of age, with less than 10 years of experience and those with young children. Stress, anxiety and depression, as well as the emotional impact of traumatic events, have been described as related psychological factors. CONCLUSION: Although extrinsic work factors such as shifts, workload, pay and interpersonal relationships increase burnout, intrinsic factors such as lack of autonomy and recognition are the main factors related to it. IMPACT: What problem did the study address? Burnout among healthcare workers has been recognized as a global crisis requiring urgent attention, specifically in midwives. What were the main findings? There is a persistent shortage of midwives that is attributed in part to chronic retention difficulties related to job burnout expressed by these professionals. Where and on whom will the research have an impact? We seek to address the paucity of research on burnout in midwives in the current crisis in the profession. Work factors such as lack of autonomy or recognition in the profession carry an associated risk of burnout and job attrition. Understanding the factors that contribute to burnout will enable healthcare organizations to reduce the current problem. REPORTING METHOD: PREFERRED: Reporting items for systematic review and meta-analyses (PRISMA). PATIENT OF PUBLIC CONTRIBUTION: No patient or public contribution.

Revolution and Resistance in the Desert: The Guggenheim System's Impact on Nitrate Mining and Society in Atacama, 1926-31.

In 1926, during an economic crisis that severely impacted the mining industry, Guggenheim Brothers, the Guggenheim family business, implemented a new technological system to extract saltpeter from the Atacama Desert in northern Chile. Known as the Guggenheim system, this cutting-edge technological innovation had a significant impact on regional society and facilitated the introduction of Chilean saltpeter into the global fertilizer market. For this system to succeed, however, it had to incorporate a sociopolitical strategy based on a highly hierarchical and well-controlled labor force. Through their political and cultural influence in the region, the Guggenheim family's industry transformed a remote area into a state periphery, creating new ways of inhabiting the desert within a strict framework in which workers' lives were regulated by company-imposed labor discipline. With more political power than the state, the Guggenheim family sought to suppress any social agency deemed dangerous to the production of saltpeter.

The romantic relationships of adopted adolescents.

INTRODUCTION: There is a gap in the literature on the romantic relationships of adopted adolescents. To address this issue, the present study has three aims: (1) to explore differences between adopted and non-adopted adolescents in terms of their involvement in and the length of their romantic relationships; (2) to explore the quality of these relationships; and (3) to analyze associations between affective relationships and well-being in both groups. METHOD: The sample comprised 276 adopted (64.5% girls; mean age 16.3 years, 73.9% international adoptees) and 276 non-adopted (48.3% girls; mean age 16.3 years) adolescents, all of whom participated in the Spanish Health Behaviour in School-aged Children survey. RESULTS: Similar romantic relationship rates and lengths were found among adoptees and non-adoptees, as well as between international and domestic adoptees. Adoptees reported more emotional support and conflicts in their romantic relationships than their non-adopted peers. Finally, associations between the quality of the romantic relationships and well-being were similar for both groups, with more conflicts being linked to lower levels of well-being, and more emotional support and affection correlating with higher levels of well-being. DISCUSSION: The data suggest more similarities than differences between adopted and non-adopted adolescents. However, although this indicates that romantic relationships are yet another example of recovery for adopted boys and girls, further research is required, with larger and more diverse samples from multiple countries, to explore the differences observed in more detail.

ER-associated degradation adapter Sel1L is required for CD8+ T cell function and memory formation following acute viral infection.

The maintenance of antigen-specific CD8+ T cells underlies the efficacy of vaccines and immunotherapies. Pathways contributing to CD8+ T cell loss are not completely understood. Uncovering the pathways underlying the limited persistence of CD8+ T cells would be of significant benefit for developing novel strategies of promoting T cell persistence. Here, we demonstrate that murine CD8+ T cells experience endoplasmic reticulum (ER) stress following activation and that the ER-associated degradation (ERAD) adapter Sel1L is induced in activated CD8+ T cells. Sel1L loss limits CD8+ T cell function and memory formation following acute viral infection. Mechanistically, Sel1L is required for optimal bioenergetics and c-Myc expression. Finally, we demonstrate that human CD8+ T cells experience ER stress upon activation and that ER stress is negatively associated with improved T cell functionality in T cell-redirecting therapies. Together, these results demonstrate that ER stress and ERAD are important regulators of T cell function and persistence.

Deconstructing cancer with precision genome editing.

Recent advances in genome editing technologies are allowing investigators to engineer and study cancer-associated mutations in their endogenous genetic contexts with high precision and efficiency. Of these, base editing and prime editing are quickly becoming gold-standards in the field due to their versatility and scalability. Here, we review the merits and limitations of these precision genome editing technologies, their application to modern cancer research, and speculate how these could be integrated to address future directions in the field.

The Soteria project recounted by Mosher and its clinical resonances today.

BACKGROUND: The American psychiatrist Loren R. Mosher has passed to posterity as an eager proponent of a psychosocial approach to psychosis. The best example of this is the Soteria project that he founded in San Jose, California, in the 1970s. The contribution of Alma Zito Menn, ACSW, also merits attention as project director of Soteria and for her links to the Mental Research Institute, Palo Alto. She was later replaced as program director by Voyce Hendrix, LCSW, when she turned to other preoccupations linked to the grant continuation of Soteria. Equally, the nonprofessional staff of the facility should receive appreciation. AIM/OBJECTIVE: Bearing this in mind, the main aim of this paper is to reflect upon the Soteria project, giving voice to Mosher himself, while simultaneously connecting his ideas with other empirical works that have been published on this topic in recent decades. METHODS: Using a selection from the extant literature assessing the implementation and outcomes of adapting Soteria-elements to different settings, I present here provisional results obtained from current research. First, I expound what Mosher hoped to achieve in creating Soteria and why it worked. In this respect, I go beyond what is commonly reported in scholarly works, where the Soteria project is considered without paying too great attention to its main architect, as if the project could be separated from the man who created it. RESULTS AND CONCLUSIONS: As I have corroborated here, there is today growing and promising scientific evidence validating the principles of the Soteria project. Undoubtedly, this would not have been possible without the pioneering work of Mosher, who, imbued with the tenets of interpersonal phenomenology, shook the psychiatric establishment, leading others to follow the path that he had begun.

High-throughput evaluation of genetic variants with prime editing sensor libraries.

Tumor genomes often harbor a complex spectrum of single nucleotide alterations and chromosomal rearrangements that can perturb protein function. Prime editing has been applied to install and evaluate genetic variants, but previous approaches have been limited by the variable efficiency of prime editing guide RNAs. Here we present a high-throughput prime editing sensor strategy that couples prime editing guide RNAs with synthetic versions of their cognate target sites to quantitatively assess the functional impact of endogenous genetic variants. We screen over 1,000 endogenous cancer-associated variants of TP53-the most frequently mutated gene in cancer-to identify alleles that impact p53 function in mechanistically diverse ways. We find that certain endogenous TP53 variants, particularly those in the p53 oligomerization domain, display opposite phenotypes in exogenous overexpression systems. Our results emphasize the physiological importance of gene dosage in shaping native protein stoichiometry and protein-protein interactions, and establish a framework for studying genetic variants in their endogenous sequence context at scale.

Expanding the Clinical Spectrum of DRP2-Associated Charcot-Marie-Tooth Disease.

BACKGROUND AND OBJECTIVES: Germline truncating variants in the DRP2 gene (encoding dystrophin-related protein 2) cause the disruption of the periaxin-DRP2-dystroglycan complex and have been linked to Charcot-Marie-Tooth disease. However, the causality and the underlying phenotype of the genetic alterations are not clearly defined. METHODS: This cross-sectional retrospective observational study includes 9 patients with Charcot-Marie-Tooth disease (CMT) with DRP2 germline variants evaluated at 6 centers throughout Spain. RESULTS: We identified 7 Spanish families with 4 different DRP2 likely pathogenic germline variants. In agreement with an X-linked inheritance, men harboring hemizygous DRP2 variants presented with an intermediate form of CMT, whereas heterozygous women were asymptomatic. Symptom onset was variable (36.6 ± 16 years), with lower limb weakness and multimodal sensory loss producing a mild-to-moderate functional impairment. Nerve echography revealed an increase in the cross-sectional area of nerve roots and proximal nerves. Lower limb muscle magnetic resonance imaging confirmed the presence of a length-dependent fatty infiltration. Immunostaining in intradermal nerve fibers demonstrated the absence of DRP2 and electron microscopy revealed abnormal myelin thickness that was also detectable in the sural nerve sections. DISCUSSION: Our findings support the causality of DRP2 pathogenic germline variants in CMT and further define the phenotype as a late-onset sensory and motor length-dependent neuropathy, with intermediate velocities and thickening of proximal nerve segments.

The role of genetics in the treatment of dystonia with deep brain stimulation: Systematic review and Meta-analysis.

BACKGROUND: Dystonia is a movement disorder characterized by sustained or intermittent muscle contractions that lead to involuntary postures or repetitive movements. Genetic mutations are being increasingly recognized as a cause of dystonia. Deep brain stimulation (DBS) is one of the limited treatment options available. However, there are varying reports on its efficacy in genetic dystonias. This systematic review of the characteristics of genetic dystonias treated with DBS and their outcomes aims to aid in the evaluation of eligibility for such treatment. METHODS: We performed a PUBMED search of all papers related to genetic dystonias and DBS up until April 2022. In addition to performing a systematic review, we also performed a meta-analysis to assess the role of the mutation on DBS response. We included cases that had a confirmed genetic mutation and DBS along with pre-and post-operative BFMDRS. RESULTS: Ninety-one reports met our inclusion criteria and from them, 235 cases were analyzed. Based on our analysis DYT-TOR1A dystonia had the best evidence for DBS response and Rapid-Onset Dystonia Parkinsonism was among the least responsive to DBS. CONCLUSION: While our report supports the role of genetics in DBS selection and response, it is limited by the rarity of the individual genetic conditions, the reliance on case reports and case series, and the limited ability to obtain genetic testing on a large scale in real-time as opposed to retrospectively as in many cases.

Metabolic reprogramming by histone deacetylase inhibition preferentially targets NRF2-activated tumors.

The interplay between metabolism and chromatin signaling is implicated in cancer progression. However, whether and how metabolic reprogramming in tumors generates chromatin vulnerabilities remain unclear. Lung adenocarcinoma (LUAD) tumors frequently harbor aberrant activation of the NRF2 antioxidant pathway, which drives aggressive and chemo-resistant disease. Using a chromatin-focused CRISPR screen, we report that NRF2 activation sensitizes LUAD cells to genetic and chemical inhibition of class I histone deacetylases (HDACs). This association is observed across cultured cells, mouse models, and patient-derived xenografts. Integrative epigenomic, transcriptomic, and metabolomic analysis demonstrates that HDAC inhibition causes widespread redistribution of H4ac and its reader protein, which transcriptionally downregulates metabolic enzymes. This results in reduced flux into amino acid metabolism and de novo nucleotide synthesis pathways that are preferentially required for the survival of NRF2-active cancer cells. Together, our findings suggest NRF2 activation as a potential biomarker for effective repurposing of HDAC inhibitors to treat solid tumors.

Somatic mouse models of gastric cancer reveal genotype-specific features of metastatic disease.

Metastatic gastric carcinoma is a highly lethal cancer that responds poorly to conventional and molecularly targeted therapies. Despite its clinical relevance, the mechanisms underlying the behavior and therapeutic response of this disease are poorly understood owing, in part, to a paucity of tractable models. Here we developed methods to somatically introduce different oncogenic lesions directly into the murine gastric epithelium. Genotypic configurations observed in patients produced metastatic gastric cancers that recapitulated the histological, molecular and clinical features of all nonviral molecular subtypes of the human disease. Applying this platform to both wild-type and immunodeficient mice revealed previously unappreciated links between the genotype, organotropism and immune surveillance of metastatic cells, which produced distinct patterns of metastasis that were mirrored in patients. Our results establish a highly portable platform for generating autochthonous cancer models with flexible genotypes and host backgrounds, which can unravel mechanisms of gastric tumorigenesis or test new therapeutic concepts.

A prime editor mouse to model a broad spectrum of somatic mutations in vivo.

Genetically engineered mouse models only capture a small fraction of the genetic lesions that drive human cancer. Current CRISPR-Cas9 models can expand this fraction but are limited by their reliance on error-prone DNA repair. Here we develop a system for in vivo prime editing by encoding a Cre-inducible prime editor in the mouse germline. This model allows rapid, precise engineering of a wide range of mutations in cell lines and organoids derived from primary tissues, including a clinically relevant Kras mutation associated with drug resistance and Trp53 hotspot mutations commonly observed in pancreatic cancer. With this system, we demonstrate somatic prime editing in vivo using lipid nanoparticles, and we model lung and pancreatic cancer through viral delivery of prime editing guide RNAs or orthotopic transplantation of prime-edited organoids. We believe that this approach will accelerate functional studies of cancer-associated mutations and complex genetic combinations that are challenging to construct with traditional models.

Laing and Rogers in London: An unfortunate meeting or something else?

BACKGROUND: In the summer of 1978 a large 1-day event was scheduled to take place in the Grand Ballroom at the Hilton Hotel in Park Lane, London between the psychotherapists Carl R. Rogers (1902-1987), and his associates, and Ronald D. Laing (1927-1989) and his group. From among all the eyewitness accounts of that meeting, I have found only the testimonies of Maureen O'Hara, Ian Cunningham, Charles Elliot, and Emmy van Deurzen. According to O'Hara, Laing behaved in a rude, impolite, and aggressive way toward his American colleague Rogers. For his part, Cunningham says that Rogers came over as he had expected: a genuinely nice, caring, humane person. Laing, though, was even more impressive in person than in his books. Similarly, Elliot observes that Laing and Rogers held a genuine encounter, one in which both sat like two real mutually respecting persons who asked each other questions, while the perspective of van Deurzen is more in line with that of O'Hara than that of Elliot. AIMS: Taking into account the different versions given on the Laing-Rogers event, I will analyze whether this encounter was only an unfortunate meeting or something else. METHODS: Narrative review; combining eyewitness accounts with the few sources found in the literature on this topic. RESULTS/CONCLUSIONS: As I will show here, all these accounts taken jointly paint a picture of Laing as a brilliant clinician and as a terrible man. Without exculpating Laing for committing all sorts of mischief, I will offer a tentative account of his behavior sustained by his own psychic dynamics. In doing so, I will attempt to explain why Laing reacted in so censurable a way, going beyond Thomas S. Szasz's (1920-2012) condemnation in his essay on antipsychiatry, which gives credence only to O'Hara's version without quoting more sources or posing more questions.