Dissecting the treatment-naive ecosystem of human melanoma brain metastasis.

Melanoma brain metastasis (MBM) frequently occurs in patients with advanced melanoma; yet, our understanding of the underlying salient biology is rudimentary. Here, we performed single-cell/nucleus RNA-seq in 22 treatment-naive MBMs and 10 extracranial melanoma metastases (ECMs) and matched spatial single-cell transcriptomics and T cell receptor (TCR)-seq. Cancer cells from MBM were more chromosomally unstable, adopted a neuronal-like cell state, and enriched for spatially variably expressed metabolic pathways. Key observations were validated in independent patient cohorts, patient-derived MBM/ECM xenograft models, RNA/ATAC-seq, proteomics, and multiplexed imaging. Integrated spatial analyses revealed distinct geography of putative cancer immune evasion and evidence for more abundant intra-tumoral B to plasma cell differentiation in lymphoid aggregates in MBM. MBM harbored larger fractions of monocyte-derived macrophages and dysfunctional TOX+CD8+ T cells with distinct expression of immune checkpoints. This work provides comprehensive insights into MBM biology and serves as a foundational resource for further discovery and therapeutic exploration.

Collagenolysis-dependent DDR1 signalling dictates pancreatic cancer outcome.

Pancreatic ductal adenocarcinoma (PDAC) is a highly desmoplastic, aggressive cancer that frequently progresses and spreads by metastasis to the liver1. Cancer-associated fibroblasts, the extracellular matrix and type I collagen (Col I) support2,3 or restrain the progression of PDAC and may impede blood supply and nutrient availability4. The dichotomous role of the stroma in PDAC, and the mechanisms through which it influences patient survival and enables desmoplastic cancers to escape nutrient limitation, remain poorly understood. Here we show that matrix-metalloprotease-cleaved Col I (cCol I) and intact Col I (iCol I) exert opposing effects on PDAC bioenergetics, macropinocytosis, tumour growth and metastasis. Whereas cCol I activates discoidin domain receptor 1 (DDR1)-NF-κB-p62-NRF2 signalling to promote the growth of PDAC, iCol I triggers the degradation of DDR1 and restrains the growth of PDAC. Patients whose tumours are enriched for iCol I and express low levels of DDR1 and NRF2 have improved median survival compared to those whose tumours have high levels of cCol I, DDR1 and NRF2. Inhibition of the DDR1-stimulated expression of NF-κB or mitochondrial biogenesis blocks tumorigenesis in wild-type mice, but not in mice that express MMP-resistant Col I. The diverse effects of the tumour stroma on the growth and metastasis of PDAC and on the survival of patients are mediated through the Col I-DDR1-NF-κB-NRF2 mitochondrial biogenesis pathway, and targeting components of this pathway could provide therapeutic opportunities.

Opposing roles of hepatic stellate cell subpopulations in hepatocarcinogenesis.

Hepatocellular carcinoma (HCC), the fourth leading cause of cancer mortality worldwide, develops almost exclusively in patients with chronic liver disease and advanced fibrosis1,2. Here we interrogated functions of hepatic stellate cells (HSCs), the main source of liver fibroblasts3, during hepatocarcinogenesis. Genetic depletion, activation or inhibition of HSCs in mouse models of HCC revealed their overall tumour-promoting role. HSCs were enriched in the preneoplastic environment, where they closely interacted with hepatocytes and modulated hepatocarcinogenesis by regulating hepatocyte proliferation and death. Analyses of mouse and human HSC subpopulations by single-cell RNA sequencing together with genetic ablation of subpopulation-enriched mediators revealed dual functions of HSCs in hepatocarcinogenesis. Hepatocyte growth factor, enriched in quiescent and cytokine-producing HSCs, protected against hepatocyte death and HCC development. By contrast, type I collagen, enriched in activated myofibroblastic HSCs, promoted proliferation and tumour development through increased stiffness and TAZ activation in pretumoural hepatocytes and through activation of discoidin domain receptor 1 in established tumours. An increased HSC imbalance between cytokine-producing HSCs and myofibroblastic HSCs during liver disease progression was associated with increased HCC risk in patients. In summary, the dynamic shift in HSC subpopulations and their mediators during chronic liver disease is associated with a switch from HCC protection to HCC promotion.

A molecular single-cell lung atlas of lethal COVID-19.

Respiratory failure is the leading cause of death in patients with severe SARS-CoV-2 infection1,2, but the host response at the lung tissue level is poorly understood. Here we performed single-nucleus RNA sequencing of about 116,000 nuclei from the lungs of nineteen individuals who died of COVID-19 and underwent rapid autopsy and seven control individuals. Integrated analyses identified substantial alterations in cellular composition, transcriptional cell states, and cell-to-cell interactions, thereby providing insight into the biology of lethal COVID-19. The lungs from individuals with COVID-19 were highly inflamed, with dense infiltration of aberrantly activated monocyte-derived macrophages and alveolar macrophages, but had impaired T cell responses. Monocyte/macrophage-derived interleukin-1ß and epithelial cell-derived interleukin-6 were unique features of SARS-CoV-2 infection compared to other viral and bacterial causes of pneumonia. Alveolar type 2 cells adopted an inflammation-associated transient progenitor cell state and failed to undergo full transition into alveolar type 1 cells, resulting in impaired lung regeneration. Furthermore, we identified expansion of recently described CTHRC1+ pathological fibroblasts3 contributing to rapidly ensuing pulmonary fibrosis in COVID-19. Inference of protein activity and ligand-receptor interactions identified putative drug targets to disrupt deleterious circuits. This atlas enables the dissection of lethal COVID-19, may inform our understanding of long-term complications of COVID-19 survivors, and provides an important resource for therapeutic development.

A Therapeutically Targetable TAZ-TEAD2 Pathway Drives the Growth of Hepatocellular Carcinoma via ANLN and KIF23.

BACKGROUND & AIMS: Despite recent progress, long-term survival remains low for hepatocellular carcinoma (HCC). The most effective HCC therapies target the tumor immune microenvironment (TIME), and there are almost no therapies that directly target tumor cells. Here, we investigated the regulation and function of tumor cell-expressed Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) in HCC. METHODS: HCC was induced in mice by Sleeping Beauty-mediated expression of MET, CTNNB1-S45Y, or TAZ-S89A, or by diethylnitrosamine plus CCl4. Hepatocellular TAZ and YAP were deleted in floxed mice via adeno-associated virus serotype 8-mediated expression of Cre. TAZ target genes were identified from RNA sequencing, confirmed by chromatin immunoprecipitation, and evaluated in a clustered regularly interspaced short palindromic repeats interference (CRISPRi) screen. TEA domain transcription factors (TEADs), anillin (ANLN), Kif23, and programmed cell death protein ligand 1 were knocked down by guide RNAs in dead clustered regularly interspaced short palindromic repeats-associated protein 9 (dCas9) knock-in mice. RESULTS: YAP and TAZ were up-regulated in murine and human HCC, but only deletion of TAZ consistently decreased HCC growth and mortality. Conversely, overexpression of activated TAZ was sufficient to trigger HCC. TAZ expression in HCC was regulated by cholesterol synthesis, as demonstrated by pharmacologic or genetic inhibition of 3-hydroxy-3-methylglutaryl- coenzyme A reductase (HMGCR), farnesyl pyrophosphate synthase, farnesyl-diphosphate farnesyltransferase 1 (FDFT1), or sterol regulatory element-binding protein 2 (SREBP2). TAZ- and MET/CTNNB1-S45Y-driven HCC required the expression of TEAD2 and, to a lesser extent, TEAD4. Accordingly, TEAD2 displayed the most profound effect on survival in patients with HCC. TAZ and TEAD2 promoted HCC via increased tumor cell proliferation, mediated by TAZ target genes ANLN and kinesin family member 23 (KIF23). Therapeutic targeting of HCC, using pan-TEAD inhibitors or the combination of a statin with sorafenib or anti-programmed cell death protein 1, decreased tumor growth. CONCLUSIONS: Our results suggest the cholesterol-TAZ-TEAD2-ANLN/KIF23 pathway as a mediator of HCC proliferation and tumor cell-intrinsic therapeutic target that could be synergistically combined with TIME-targeted therapies.

Histone acetylation of bile acid transporter genes plays a critical role in cirrhosis.

BACKGROUND & AIMS: Owing to the lack of genetic animal models that adequately recreate key clinical characteristics of cirrhosis, the molecular pathogenesis of cirrhosis has been poorly characterized, and treatments remain limited. Hence, we aimed to better elucidate the pathological mechanisms of cirrhosis using a novel murine model. METHODS: We report on the first murine genetic model mimicking human cirrhosis induced by hepatocyte-specific elimination of microspherule protein 1 (MCRS1), a member of non-specific lethal (NSL) and INO80 chromatin-modifier complexes. Using this genetic tool with other mouse models, cell culture and human samples, combined with quantitative proteomics, single nuclei/cell RNA sequencing and chromatin immunoprecipitation assays, we investigated mechanisms of cirrhosis. RESULTS: MCRS1 loss in mouse hepatocytes modulates the expression of bile acid (BA) transporters - with a pronounced downregulation of Na+-taurocholate cotransporting polypeptide (NTCP) - concentrating BAs in sinusoids and thereby activating hepatic stellate cells (HSCs) via the farnesoid X receptor (FXR), which is predominantly expressed in human and mouse HSCs. Consistently, re-expression of NTCP in mice reduces cirrhosis, and genetic ablation of FXR in HSCs suppresses fibrotic marks in mice and in vitro cell culture. Mechanistically, deletion of a putative SANT domain from MCRS1 evicts histone deacetylase 1 from its histone H3 anchoring sites, increasing histone acetylation of BA transporter genes, modulating their expression and perturbing BA flow. Accordingly, human cirrhosis displays decreased nuclear MCRS1 and NTCP expression. CONCLUSIONS: Our data reveal a previously unrecognized function of MCRS1 as a critical histone acetylation regulator, maintaining gene expression and liver homeostasis. MCRS1 loss induces acetylation of BA transporter genes, perturbation of BA flow, and consequently, FXR activation in HSCs. This axis represents a central and universal signaling event in cirrhosis, which has significant implications for cirrhosis treatment. LAY SUMMARY: By genetic ablation of MCRS1 in mouse hepatocytes, we generate the first genetic mouse model of cirrhosis that recapitulates human features. Herein, we demonstrate that the activation of the bile acid/FXR axis in liver fibroblasts is key in cirrhosis development.

Meditation, well-being and cognition in heartfulness meditators - A pilot study.

Meditative practices can vary considerably in technique as well as their effects. Heartfulness is a popular meditation technique that includes in its repertoire, a unique passive form of meditation. We carried out a pilot study recruiting male heartfulness meditators (proficient n = 24, with 6-28 years of meditation experience; novice n = 24, 5 to 16 months of experience) and subsequently recruited matched controls (n = 15). We examined well-being, and carried out high-density EEG recordings to examine indices of meditation and cognition in these groups. Well-being scores were significantly higher for the proficient meditators as compared to novice and intermediate for the controls. We did not find any group differences in cognitive processing. During meditation, enhanced occipital gamma was found in proficient meditators as compared to controls. We discuss the findings from this pilot study and suggest avenues for future research.

Beyond Hypnograms: Assessing Sleep Stability Using Acoustic and Electrical Stimulation.

OBJECTIVE: Conventional polysomnographic recordings reflect brain dynamics associated with sleep architecture. We hypothesized that noninvasive tools like transcranial alternating current stimulation (tACS) and acoustic stimulation (for generating event related potentials [ERPs]) would help to predict sleep stability and provide a window to actively assess brain activity during sleep. MATERIALS AND METHODS: Twelve healthy male volunteers participated in the multiple whole-night polysomnography (PSG) recording protocol. Acoustic tones (100 msec duration) were presented throughout night to evaluate ERP during sleep. Furthermore, 30 sec tACS were presented during nonrapid eye movement (NREM) and rapid eye movement (REM) sleep on subsequent two nights without disturbing the subject's sleep. For ERP analysis, event-locked artifact-free epochs from each sleep stage were averaged separately. For tACS analysis, 30 sec prestimulus and poststimulus artifact-free EEG epochs were subjected to bootstrapping-based comparison of power spectral values. RESULTS: Acoustic stimulation generated sleep stage-dependent ERP components (N350, N550, and P900) in all participants. The tACS stimulation during NREM sleep (0.75 Hz) increased parietal delta power but decreased frontocentral theta and increased frontal gamma power when delivered during REM sleep (40Hz). These interventions provide details on sleep stability as larger N550-P900 ERP-complex correlated with lower NREM disruptions (Spearman's rho = -0.553; p = 0.049; n = 10) and tACS-related theta power perturbation with higher REM disruptions (Spearman's rho = 0.734; p = 0.030; n = 7). CONCLUSIONS: Noninvasive brain stimulation approaches such as sleep ERP and sleep tACS are reliable tools to evaluate sleep stability during NREM and REM sleep, respectively, but more large-sample studies are warranted.

Just a minute meditation: Rapid voluntary conscious state shifts in long term meditators.

Meditation induces a modified state of consciousness that remains under voluntary control. Can meditators rapidly and reversibly bring about mental state changes on demand? To check, we carried out 128 channel EEG recordings on Brahma Kumaris Rajayoga meditators (36 long term: median 14240h meditation; 25 short term: 1095h) and controls (25) while they tried to switch every minute between rest and meditation states in different conditions (eyes open and closed; before and after an engaging task). Long term meditators robustly shifted states with enhanced theta power (4-8Hz) during meditation. Short term meditators had limited ability to shift between states and showed increased lower alpha power (8-10Hz) during eyes closed meditation only when pre and post task data were combined. Controls could not shift states. Thus trained beginners can reliably meditate but it takes long term practice to exercise more refined control over meditative states.

Comparison of outcomes of different Graf grades of developmental dysplasia of the hip in infants treated with Tubingen splint versus Pavlik harness - A systematic review.

This systematic review was designed to compare the outcomes of the two braces against each other classified by the Graf method. The databases sources included PubMed, Embase, and Google Scholar. The keywords included "DDH Tubingen versus Pavlik" and Tubingen and Pavlik separately. Included papers provided specific data regarding success and failure rate, avascular necrosis (AVN), duration, and age of intervention. The excluded studies discussed surgeries, diagnosis and mechanism, and ones that weren't in English. Total of 20 papers were included, resulting in 1243 Tubingen and 420 Pavlik samples. It was seen that the Tubingen splint had a statistically significant greater success rate and lower failure rate for Graf 2, D, and 3 hips, while both braces were not very successful for Graf 4 at success rates less than 60 %. Tubingen also had a lower incidence of AVN. Both braces shared similar ages of intervention, duration, and time per day. Both braces are very comparable to each other, each having better success rates for lower Graf grades, which points to the importance of bracing earlier to improve the success rates. The Tubingen splint had a higher success rate, lower failure rate, and lower AVN rate compared to the Pavlik harness. This points to the Tubingen splint potentially being the preferred option for bracing in infants.

Cement Embolism After Kyphoplasty.

Kyphoplasty is used for the treatment of vertebral compression fractures. The procedure involves inflating a balloon at the compression site; then, polymethylmethacrylate (PMMA) cement is added into the space created by the balloon, where it polymerizes, achieving stabilization, with possible expansion of the vertebral angle. The process is guided by X-rays. Complications are rare, especially when compared to vertebroplasty, and one rare complication is pulmonary cement embolism (PCE). Although many cases are likely undetected due to a lack of symptoms, symptomatic cases require treatment, as they can sometimes prove fatal. We present a case of a patient who underwent kyphoplasty and later presented with a PCE. The PCE was diagnosed using X-rays and computed tomography (CT).

Pediatric central nervous system (CNS) neuroblastoma: A case report.

Background: Neuroblastomas are rare tumors activated by the FoxR2 gene commonly found in pediatric patients. Due to the novelty of these tumors, there is no standard diagnostic profile. However, they have been found to express Olig2, MAP2, SOX10, ANKRD55, and synaptophysin, and they can be identified with magnetic resonance imaging (MRI). Treatment with chemotherapy combined with stem cell rescue and craniospinal irradiation can improve non-infant patient outcomes. Case Description: We report a case of a 2-year-old patient who was diagnosed with a neuroblastoma through MRI imaging and pathology that confirmed FoxR2 gene activation. The tumor was successfully removed. However, the tumor was not high-grade like most FoxR2 neuroblastomas. Conclusion: The unusual presentation of a low-grade FoxR2 neuroblastoma demonstrates the necessity to conduct further research into the characteristics of these tumors.

Impacts of Omega-3 Fatty Acids, Natural Elixirs for Neuronal Health, on Brain Development and Functions.

Omega-3 fatty acids play a seminal role in maintaining the structural and functional integrity of the nervous system. These specialized molecules function as precursors for many lipid-based biological messengers. Also, studies suggest the role of these fatty acids in regulating healthy sleep cycles, cognitive ability, brain development, etc. Dietary intake of essential poly unsaturated fatty acids (PUFA) such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are foundational to the optimal working of the nervous system. Besides regulating health, these biomolecules have great therapeutic value in treating several diseases, particularly nervous system diseases and disorders. Many recent studies conclusively demonstrated the beneficial effects of Omega-3 fatty acids in treating depression, neuropsychiatric disorders, neurodegenerative disorders, neurochemical disorders, and many other illnesses associated with the nervous system. This chapter summates the multifaceted role of poly unsaturated fatty acids, especially Omega-3 fatty acids (EPA and DHA), in the neuronal health and functioning. The importance of dietary intake of these essential fatty acids, their recommended dosages, bioavailability, the mechanism of their action, and therapeutic values are extensively discussed.

Exploring the potential of telemedicine for improved primary healthcare in India: a comprehensive review.

Telemedicine is a promising solution to the challenges of delivering equitable and quality primary healthcare, especially in LMICs. This review evaluated peer-reviewed literature on telehealth interventions in Indian primary care published from Jan 1, 2011 to Dec 31, 2021, from PubMed, Scopus, TRIP, Google Scholar, Indian Kanoon, and Cochrane database The majority of Indian studies focus on key health issues like maternal and child health, mental health, diabetes, infectious diseases, and hypertension, mainly through patient education, monitoring, and diagnostics. Yet, there's a lack of research on telemedicine's cost-effectiveness, communication among providers, and the role of leadership in its quality and accessibility. The current research has gaps, including small sample sizes and inconsistent methodologies, which hamper the evaluation of telemedicine's effectiveness. India's varied healthcare landscape, technological limitations, and social factors further challenge telemedicine's adoption. Despite regulatory efforts, issues like the digital divide and data privacy persist. Addressing these challenges with a context-aware, technologically driven approach is crucial for enhancing healthcare through telemedicine in India.

Purpose in life as a resilience factor for brain health: diffusion MRI findings from the Midlife in the U.S. study.

Introduction: A greater sense of purpose in life is associated with several health benefits relevant for active aging, but the mechanisms remain unclear. We evaluated if purpose in life was associated with indices of brain health. Methods: We examined data from the Midlife in the United States (MIDUS) Neuroscience Project. Diffusion weighted magnetic resonance imaging data (n=138; mean age 65.2 years, age range 48-95; 80 females; 37 black, indigenous, and people of color) were used to estimate microstructural indices of brain health such as axonal density, and axonal orientation. The seven-item purpose in life scale was used. Permutation analysis of linear models was used to examine associations between purpose in life scores and the diffusion metrics in white matter and in the bilateral hippocampus, adjusting for age, sex, education, and race. Results and discussion: Greater sense of purpose in life was associated with brain microstructural features consistent with better brain health. Positive associations were found in both white matter and the right hippocampus, where multiple convergent associations were detected. The hippocampus is a brain structure involved in learning and memory that is vulnerable to stress but retains the capacity to grow and adapt through old age. Our findings suggest pathways through which an enhanced sense of purpose in life may contribute to better brain health and promote healthy aging. Since purpose in life is known to decline with age, interventions and policy changes that facilitate a greater sense of purpose may extend and improve the brain health of individuals and thus improve public health.

Role of oxygenation devices in alleviating the oxygen crisis in India.

There has been an unprecedented increase in global demand for medical oxygen equipment to solve the acute oxygen shortages caused by SARS-CoV-2 infection. The study aims to assess the value of improved access and use of Oxygen Concentrators (OCs) and cylinders during the COVID-19 pandemic in India. This evaluation is relevant to strengthening health systems in many resource-constrained Low- and Middle-Income Country (LMIC) settings. Using a Probability Proportional to Size (PPS) sampling method, primary surveys were conducted in 450 health facilities across 21 states in India. The primary outcomes measured were self-reported utility of oxygenation devices in meeting the oxygen demand in the short-run and long-run utility of devices compared to the pre-oxygen-devices-distribution-period. We perform bivariate and multivariate regression analyses. Around 53-54% of surveyed facilities reported that the distributed oxygenation devices helped meet oxygen demand in the short run and are expected to increase their long-run capacity to admit non-COVID patients with oxygen needs. The timely availability of technicians was associated with meeting oxygen demand using the additional oxygenation devices at the facilities. Facilities that increased the number of staff members who were able to administer oxygen devices were at higher odds of reducing the administrative load on their staff to organize oxygen support in the long run. Hospital infrastructure was also associated with long-run outcomes. We find that oxygenation devices such as cylinders and OCs were useful in addressing the oxygen demand during the COVID-19-related oxygen emergency. Overall production of oxygen to meet the demands and investments in training biomedical engineers/technicians to administer oxygen could help save lives.

Differences in brain connectivity of meditators during assessing neurocognition via gamified experimental logic task: A machine learning approach.

Meditation practices increase attention, memory, and self-awareness. The neuroscientific study of meditation has helped gain useful insights into the functional changes in the brain. In this study, we have assessed the performance of meditators with different years of practice while performing an engaging task rather than studying the meditation practice itself. This task helps assess many neural processes simultaneously and represents task performance in presence of multiple audio-visual distractors as in a real-life scenario. The long-term practice of meditation could bring neuroplastic changes in the way cognitive processing is carried out. It could be conscious and effortful in short-term practitioners and relatively unconscious and effortless in long-term practitioners. Our goal is to understand if it is possible to differentiate between long-term and short-term meditators solely based on their cognitive processing. A group of proficient Rajayoga meditators from the Brahma Kumaris were recruited based on their meditation experience-Long-Term Practitioners (n = 12, mean 13,596 h) and Short-Term Practitioners (n = 10, mean 1095 h). A task-based functional Magnetic Resonance Imaging was acquired while the subjects performed the task. Functional Connectivity Analysis was performed to derive the correlation measures to be used as features for classification. Five supervised Machine Learning algorithms Logistic Regression, Support Vector Machine, Decision Tree, Random Forest, and Gradient Boosted Tree were used for classification. Among all the classifiers Gradient Boosted Tree performed the best with an accuracy of 77% when all the four Functional Connectivity Metrics were used. Connectivity in visual areas, cerebellum, left rostral prefrontal cortex, and middle frontal gyrus was found to be higher in long-term meditators. Such a classification demonstrates that long-term meditation practice brings about neuroplastic changes that influence cognitive processing.

Altered Developmental Trajectory in Male and Female Rats in a Prenatal Valproic Acid Exposure Model of Autism Spectrum Disorder.

Early motor and sensory developmental delays precede Autism Spectrum Disorder (ASD) diagnosis and may serve as early indicators of ASD. The literature on sensorimotor development in animal models is sparse, male centered, and has mixed findings. We characterized early development in a prenatal valproic acid (VPA) model of ASD and found sex-specific developmental delays in VPA rats. We created a developmental composite score combining 15 test readouts, yielding a reliable gestalt measure spanning physical, sensory, and motor development, that effectively discriminated between VPA and control groups. Considering the heterogeneity in ASD phenotype, the developmental composite offers a robust metric that can enable comparison across different animal models of ASD and can serve as an outcome measure for early intervention studies.