Paired CRISPR screens to map gene regulation in and .

Recent massively-parallel approaches to decipher gene regulatory circuits have focused on the discovery of either -regulatory elements (CREs) or -acting factors. Here, we develop a scalable approach that pairs - and -regulatory CRISPR screens to systematically dissect how the key immune checkpoint is regulated. In human pancreatic ductal adenocarcinoma (PDAC) cells, we tile the locus using ∼25,000 CRISPR perturbations in constitutive and IFNγ-stimulated conditions.

Transcriptome-scale RNA-targeting CRISPR screens reveal essential lncRNAs in human cells.

Mammalian genomes host a diverse array of RNA that includes protein-coding and noncoding transcripts. However, the functional roles of most long noncoding RNAs (lncRNAs) remain elusive. Using RNA-targeting CRISPR-Cas13 screens, we probed how the loss of ∼6,200 lncRNAs impacts cell fitness across five human cell lines and identified 778 lncRNAs with context-specific or broad essentiality.

Ruthenium-catalyzed Heck coupling of 3-arylidene-oxindoles with alkenes: a facile synthesis of 3-allylidene-2(3)-oxindoles.

A simple and efficient Ru(II)-catalyzed olefination of 3-(arylbenzylidene)indolin-2-ones with alkenes is described. This is an atom and step-economical strategy with a wide substrate scope, good functional group tolerance, and suitability for gram scale synthesis. A plausible mechanism is also proposed for this synthetic transformation involving the formation of a 5-membered ruthenacycle and insertion of the alkene followed by β-hydride elimination to deliver the desired product.

Comparing single- and multi-post labeling delays for the measurements of resting cerebral and hippocampal blood flow for cerebrovascular testing in midlife adults.

Objectives: To assess the reliability and validity of measuring resting cerebral blood flow (CBF) and hippocampal CBF using a single-post-labeling delay (PLD) and a multi-PLD pseudo-continuous arterial spin labeling (pCASL) protocol for cerebrovascular reactivity (CVR) testing.

Methods: 25 healthy, midlife adults (57 ± 4 years old) were imaged in a Siemens Prisma 3T magnetic resonance imaging (MRI) scanner. Resting CBF and hippocampal CBF were assessed using two pCASL protocols, our modified single-PLD protocol (pCASL-MOD) to accommodate the needs for CVR testing and the multi-PLD Human Connectome Project (HCP) Lifespan protocol to serve as the reference control (pCASL-HCP).

Effectiveness of mental health literacy intervention and contact-based education on knowledge, attitude, and stigma towards mental health among secondary school students in Chennai, South India: A cluster randomized controlled trial.

Background: Global trends indicate a rise in mental health disorders among youth, prompting need for effective interventions. Mental health literacy interventions have demonstrated benefits such as improved knowledge, treatment understanding, help-seeking behaviors, and stigma reduction. This study aimed to assess the impact of a comprehensive mental health literacy program on knowledge, attitudes, and stigma related to mental health among secondary school students in Chennai.

Autism-associated CHD8 controls reactive gliosis and neuroinflammation via remodeling chromatin in astrocytes.

Reactive changes of glial cells during neuroinflammation impact brain disorders and disease progression. Elucidating the mechanisms that control reactive gliosis may help us to understand brain pathophysiology and improve outcomes. Here, we report that adult ablation of autism spectrum disorder (ASD)-associated CHD8 in astrocytes attenuates reactive gliosis via remodeling chromatin accessibility, changing gene expression.

Low copy CRISPR-Cas13d mitigates collateral RNA cleavage.

While CRISPR-Cas13 systems excel in accurately targeting RNA, the potential for collateral RNA degradation poses a concern for therapeutic applications and limits broader adoption for transcriptome perturbations. We evaluate the extent to which collateral RNA cleavage occurs when Cas13d is delivered via plasmid transfection or lentiviral transduction and find that collateral activity only occurs with high levels of Cas13d expression. Using transcriptome-scale and combinatorial CRISPR pooled screens in cell lines with low-copy Cas13d, we find high on-target knockdown, without extensive collateral activity regardless of the expression level of the target gene.

Subtrochanteric femur nonunion - Chasing the elusive an analysis of two techniques to achieve union: Nail-plate fixation and plate-structural fibula graft fixation.

Introduction: The subtrochanteric region is known for its unique biomechanical properties that contribute to challenges in fracture reduction. To ensure optimal fracture healing, achieving robust mechanical stability is essential. There are very few studies in the literature describing the treatment of subtrochanteric fracture nonunion.

Non-linear transcriptional responses to gradual modulation of transcription factor dosage.

Genomic loci associated with common traits and diseases are typically non-coding and likely impact gene expression, sometimes coinciding with rare loss-of-function variants in the target gene. However, our understanding of how gradual changes in gene dosage affect molecular, cellular, and organismal traits is currently limited. To address this gap, we induced gradual changes in gene expression of four genes using CRISPR activation and inactivation.

Synergistic interactions of assorted ameliorating agents to enhance the potential of heavy metal phytoremediation.

Pollution by toxic heavy metals creates a significant impact on the biotic community of the ecosystem. Nowadays, a solution to this problem is an eco-friendly approach like phytoremediation, in which plants are used to ameliorate heavy metals. In addition, various amendments are used to enhance the potential of heavy metal phytoremediation.

Structural, surface morphological and dielectric studies of guanidinium salt incorporated poly (ethylene oxide)/poly (vinyl pyrrolidone) solid polymer electrolytes.

The research article investigates the effect of incorporating the guanidinium carbonate (GuC) salt into the poly vinylpyrrolidone (PVP) and polyethylene oxide (PEO) polymer matrix. Various weight percentages of GuC enriched PVP/PEO solid polymer electrolytes (SPEs) have been prepared by the simplest solution casting process. XRD analysis revealed that the incorporation of the GuC salt led to changes in the crystalline structure of the PVP/PEO.

Essential transcription factors for induced neuron differentiation.

Neurogenins are proneural transcription factors required to specify neuronal identity. Their overexpression in human pluripotent stem cells rapidly produces cortical-like neurons with spiking activity and, because of this, they have been widely adopted for human neuron disease models. However, we do not fully understand the key downstream regulatory effectors responsible for driving neural differentiation.

Next-generation forward genetic screens: uniting high-throughput perturbations with single-cell analysis.

Programmable genome-engineering technologies, such as CRISPR (clustered regularly interspaced short palindromic repeats) nucleases and massively parallel CRISPR screens that capitalize on this programmability, have transformed biomedical science. These screens connect genes and noncoding genome elements to disease-relevant phenotypes, but until recently have been limited to individual phenotypes such as growth or fluorescent reporters of gene expression. By pairing massively parallel screens with high-dimensional profiling of single-cell types/states, we can now measure how individual genetic perturbations or combinations of perturbations impact the cellular transcriptome, proteome, and epigenome.

Non-osseous intradural tuberculoma of the thoracic spine with compressive myelopathy.

Key Clinical Message: An uncommon form of CNS tuberculosis called non-osseous IDEM tuberculoma frequently results from paradoxical drug interactions. It should be considered one of the differentials when patients receiving ATT experience acute neurological impairment.

Abstract: Tuberculoma affecting the spinal cord is a rare condition in modern times.

LKB1 suppresses growth and promotes the internalization of EGFR through the PIKFYVE lipid kinase.

The tumor suppressor LKB1 is a serine/threonine protein kinase that is frequently mutated in human lung adenocarcinoma (LUAD). LKB1 regulates a complex signaling network that is known to control cell polarity and metabolism; however, the pathways that mediate the tumor suppressive activity of LKB1 are incompletely defined. To identify mechanisms of LKB1- mediated growth suppression we developed a spheroid-based cell culture assay to study LKB1- dependent growth.

Endogenous renal adiponectin drives gluconeogenesis through enhancing pyruvate and fatty acid utilization.

Adiponectin is a secretory protein, primarily produced in adipocytes. However, low but detectable expression of adiponectin can be observed in cell types beyond adipocytes, particularly in kidney tubular cells, but its local renal role is unknown. We assessed the impact of renal adiponectin by utilizing male inducible kidney tubular cell-specific adiponectin overexpression or knockout mice.

Heterozygous deletion of the autism-associated gene CHD8 impairs synaptic function through widespread changes in gene expression and chromatin compaction.

Whole-exome sequencing of autism spectrum disorder (ASD) probands and unaffected family members has identified many genes harboring de novo variants suspected to play a causal role in the disorder. Of these, chromodomain helicase DNA-binding protein 8 (CHD8) is the most recurrently mutated. Despite the prevalence of CHD8 mutations, we have little insight into how CHD8 loss affects genome organization or the functional consequences of these molecular alterations in neurons.

Evaluation of pharyngeal airway volume three-dimensionally in various sagittal skeletal patterns - Systematic review.

To investigate the pharyngeal airway volume in different anteroposterior skeletal malocclusions. This study was prepared according to the Cochrane criteria for creating a systematic review and meta-analysis and confirms the preferred reporting items for systematic reviews and meta-analyses (PRISMA) statement. This search was conducted to answer the patient/population, intervention, comparison and outcomes (PICO) question: To evaluate (outcome) and compare (comparison) whether there is a difference in airway volume in patients (participants) with different skeletal malocclusions diagnosed using 3D data (intervention).

Cas13d-mediated isoform-specific RNA knockdown with a unified computational and experimental toolbox.

Alternative splicing is an essential mechanism for diversifying proteins, in which mature RNA isoforms produce proteins with potentially distinct functions. Two major challenges in characterizing the cellular function of isoforms are the lack of experimental methods to specifically and efficiently modulate isoform expression and computational tools for complex experimental design. To address these gaps, we developed and methodically tested a strategy which pairs the RNA-targeting CRISPR/Cas13d system with guide RNAs that span exon-exon junctions in the mature RNA.

Rapid and Specific Detection of B. melitensis Targeting BMEI1661 Gene Using Loop-mediated Isothermal Amplification (LAMP) Combined With Lateral Flow immunoassay (LFIA).

B. melitensis is the most pathogenic zoonotic species of Brucella transmitted to animals through fetal secretions, placenta, and vaginal discharges of infected animals and humans by ingesting unpasteurized milk, dairy products, and raw meat. Early detection of B.