Understanding, assessing and treating immune, endothelial and haemostasis dysfunctions in bacterial sepsis.

The interplay between the immune system, coagulation, and endothelium is critical in regulating the host response to infection. However, in sepsis and other critical illnesses, a dysregulated immune response can lead to excessive alterations in these mechanisms, resulting in coagulopathy, endothelial dysfunction, and multi-organ dysfunction. This review aims to provide a comprehensive analysis of the pathophysiological mechanisms that govern the complex interplay between immune dysfunction, endothelial dysfunction, and coagulation in sepsis. It emphasises clinical significance, evaluation methods, and potential therapeutic interventions. Understanding these mechanisms is essential for developing effective treatments that can modulate the immune response, mitigate thrombosis, restore endothelial function, and ultimately improve patient survival.

Does sleep quality differ across political parties? Results from a survey of Arizona adults.

OBJECTIVES: In this study, we explore the relationship between political party affiliation and sleep quality since the COVID-19 pandemic. METHODS: We analyze online survey data collected for a sample of adult residents of Arizona in February and March 2023 (N = 922). We fit ordered-logistic regression models to examine how party affiliation and changes to one's personal life due to the COVID-19 pandemic are associated with the self-reported frequency of sleep difficulty. RESULTS: Compared to Republicans, Democrats and Independents report significantly worse sleep quality, net of the influence of sociodemographic controls. Additionally, having experienced major changes to one's personal life due to the COVID-19 pandemic is significantly associated with more frequent trouble sleeping for Democrats and Independents, but not for Republicans. CONCLUSIONS: We document a partisan divide in sleeping patterns among adults in a swing state and highlight an underappreciated factor contributing to sleep health amidst heightened political polarization.

High-throughput gene expression analysis with TempO-LINC sensitively resolves complex brain, lung and kidney heterogeneity at single-cell resolution.

We report the development and performance of a novel genomics platform, TempO-LINC, for conducting high-throughput transcriptomic analysis on single cells and nuclei. TempO-LINC works by adding cell-identifying molecular barcodes onto highly selective and high-sensitivity gene expression probes within fixed cells, without having to first generate cDNA. Using an instrument-free combinatorial-indexing approach, all probes within the same fixed cell receive an identical barcode, enabling the reconstruction of single-cell gene expression profiles across as few as several hundred cells and up to 100,000+ cells per run. The TempO-LINC approach is easily scalable based on the number of barcodes and rounds of barcoding performed; however, for the experiments reported in this study, the assay utilized over 5.3 million unique barcodes. TempO-LINC has a robust protocol for fixing and banking cells and displays high-sensitivity gene detection from multiple diverse sample types. We show that TempO-LINC has an observed multiplet rate of less than 1.1% and a cell capture rate of ~50%. Although the assay can accurately profile the whole transcriptome (19,683 human or 21,400 mouse genes), it can be targeted to measure only actionable/informative genes and molecular pathways of interest - thereby reducing sequencing requirements. In this study, we applied TempO-LINC to profile the transcriptomes of 89,722 cells across multiple sample types, including nuclei from mouse lung, kidney and brain tissues. The data demonstrated the ability to identify and annotate at least 50 unique cell populations and positively correlate expression of cell type-specific molecular markers within them. TempO-LINC is a robust new single-cell technology that is ideal for large-scale applications/studies across thousands of samples with high data quality.

Epitaxy of GaSe Coupled to Graphene: From In Situ Band Engineering to Photon Sensing.

2D semiconductors can drive advances in quantum science and technologies. However, they should be free of any contamination; also, the crystallographic ordering and coupling of adjacent layers and their electronic properties should be well-controlled, tunable, and scalable. Here, these challenges are addressed by a new approach, which combines molecular beam epitaxy and in situ band engineering in ultra-high vacuum of semiconducting gallium selenide (GaSe) on graphene. In situ studies by electron diffraction, scanning probe microscopy, and angle-resolved photoelectron spectroscopy reveal that atomically-thin layers of GaSe align in the layer plane with the underlying lattice of graphene. The GaSe/graphene heterostructure, referred to as 2semgraphene, features a centrosymmetric (group symmetry D3d) polymorph of GaSe, a charge dipole at the GaSe/graphene interface, and a band structure tunable by the layer thickness. The newly-developed, scalable 2semgraphene is used in optical sensors that exploit the photoactive GaSe layer and the built-in potential at its interface with the graphene channel. This proof of concept has the potential for further advances and device architectures that exploit 2semgraphene as a functional building block.

Estimation of population age structure, daily survival rates, and potential to support dengue virus transmission for Florida Keys Aedes aegypti via transcriptional profiling.

Aedes aegypti is an important vector of dengue virus and other arboviruses that affect human health. After being ingested in an infectious bloodmeal, but before being transmitted from mosquito to human, dengue virus must disseminate from the vector midgut into the hemocoel and then the salivary glands. This process, the extrinsic incubation period, typically takes 6-14 days. Since older mosquitoes are responsible for transmission, understanding the age structure of vector populations is important. Transcriptional profiling can facilitate predictions of the age structures of mosquito populations, critical for estimating their potential for pathogen transmission. In this study, we utilized a two-gene transcript model to assess the age structure and daily survival rates of three populations (Key West, Marathon, and Key Largo) of Ae. aegypti from the Florida Keys, United States, where repeated outbreaks of autochthonous dengue transmission have recently occurred. We found that Key Largo had the youngest Ae. aegypti population with the lowest daily survival rate, while Key West had the oldest population and highest survival rate. Across sites, 22.67% of Ae. aegypti females were likely old enough to transmit dengue virus (at least 15 days post emergence). Computed estimates of the daily survival rate (0.8364 using loglinear and 0.8660 using non-linear regression), indicate that dengue vectors in the region experienced relatively low daily mortality. Collectively, our data suggest that Ae. aegypti populations across the Florida Keys harbor large numbers of older individuals, which likely contributes to the high risk of dengue transmission in the area.

Discovery of Potent STT3A/B Inhibitors and Assessment of Their Multipathogen Antiviral Potential and Safety.

In the aftermath of the COVID-19 pandemic, opportunities to modulate biological pathways common to the lifecycles of viruses need to be carefully considered. N-linked glycosylation in humans is mediated exclusively by the oligosaccharyltransferase complex and is frequently hijacked by viruses to facilitate infection. As such, STT3A/B, the catalytic domain of the OST complex, became an intriguing drug target with broad-spectrum antiviral potential. However, due to the critical role N-linked glycosylation plays in a number of fundamental human processes, the toxicological ramifications of STT3A/B inhibition required attention commensurate to that given to antiviral efficacy. Herein, we describe how known STT3A/B inhibitor NGI-1 inspired the discovery of superior tool compounds which were evaluated in in vitro efficacy and translational safety (e.g., CNS, cardiovascular, liver) studies. The described learnings will appeal to those interested in the therapeutic utility of modulating N-linked glycosylation as well as the broader scientific community.

Viability of shear wave elastography to predict mechanical/ultimate failure in the anterolateral and medial collateral ligaments of the knee.

The purpose of this study was (1) to determine the utility of shear wave elastography as a predictor for the mechanical failure of superficial knee ligaments and (2) to determine the viability of shear wave elastography to assess injury risk potential. Our hypothesis was that shear wave elastography measurements of the anterolateral ligament and medial collateral ligament would directly correlate with the material properties and the mechanical failure of the ligament, serving as a prognostic measurement for injury risk. 8 cadaveric specimens were acquired, and tissue stiffness for the anterolateral ligament and medial collateral ligament were evaluated with shear wave elastography. The anterolateral ligament and medial collateral ligament were dissected and isolated for unilateral mechanical failure testing. Ultimate failure testing was performed at 100 % strain per second after 50 cycles of 3 % strain viscoelastic conditioning. Each specimen was assessed for load, displacement, and surface strain throughout failure testing. Rate of force, rate of strain development, and Young's modulus were calculated from these variables. Shear wave elastography stiffness for the anterolateral ligament correlated with mean longitudinal anterolateral ligament strain at failure (R2 = 0.853; P<0.05). Medial collateral ligament shear wave elastography calculated modulus was significantly greater than the anterolateral ligament shear wave elastography calculated modulus. Shear wave elastography currently offers limited reliability in the prediction of mechanical performance of superficial knee ligaments. The utility of shear wave elastography assessment for injury risk potential remains undetermined.

Towards a Yersinia pestis lipid A recreated in an Escherichia coli scaffold genome.

Synthetic biology and genome engineering capabilities have facilitated the utilization of bacteria for a myriad of applications, ranging from medical treatments to biomanufacturing of complex molecules. The bacterial outer membrane, specifically the lipopolysaccharide (LPS), plays an integral role in the physiology, pathogenesis, and serves as a main target of existing detection assays for Gram-negative bacteria. Here we use CRISPR/Cas9 recombineering to insert Yersinia pestis lipid A biosynthesis genes into the genome of an Escherichia coli strain expressing the lipid IVa subunit. We successfully inserted three genes: kdsD, lpxM, and lpxP into the E. coli genome and demonstrated their expression via reverse transcription PCR (RT-PCR). Despite observing expression of these genes, analytical characterization of the engineered strain's lipid A structure via MALDI-TOF mass spectrometry indicated that the Y. pestis lipid A was not recapitulated in the E. coli background. As synthetic biology and genome engineering technologies advance, novel applications and utilities for the detection and treatments of dangerous pathogens like Yersinia pestis will continue to be developed.

Labels Are For Soup Cans: How Self-Labeling as "Addicted" to Pornography Is Associated with Negative Outcomes.

Pornography viewers often report being addicted to pornography even if their behavior does not indicate such addiction. In this study with 1099 participants (52% male), we explored how the specific belief in being addicted to pornography could predict both individual and relational outcomes beyond actual pornography use and reported compulsive pornography use (i.e., considering one's use to be out of control). Using a structural equation model, our results showed that higher agreement with the pornography addiction label, without accounting for compulsivity, was associated with higher depression, suicide ideation, communication discomfort about pornography, and higher odds of having a relationship end solely because of pornography. After accounting for compulsivity, higher agreement with the pornography addiction label was only associated with higher communication discomfort about pornography and higher odds of having a relationship end solely because of pornography. This study highlights that identifying as addicted to pornography may include a stigma that is particularly detrimental to relationship outcomes.

CPT2 Deficiency Modeled in Zebrafish: Abnormal Neural Development, Electrical Activity, Behavior, and Schizophrenia-Related Gene Expression.

Carnitine palmitoyltransferase 2 (CPT2) is an inner mitochondrial membrane protein of the carnitine shuttle and is involved in the beta-oxidation of long chain fatty acids. Beta-oxidation provides an alternative pathway of energy production during early development and starvation. CPT2 deficiency is a genetic disorder that we recently showed can be associated with schizophrenia. We hypothesize that CPT2 deficiency during early brain development causes transcriptional, structural, and functional abnormalities that may contribute to a CNS environment that is susceptible to the emergence of schizophrenia. To investigate the effect of CPT2 deficiency on early vertebrate development and brain function, CPT2 was knocked down in a zebrafish model system. CPT2 knockdown resulted in abnormal lipid utilization and deposition, reduction in body size, and abnormal brain development. Axonal projections, neurotransmitter synthesis, electrical hyperactivity, and swimming behavior were disrupted in CPT2 knockdown zebrafish. RT-qPCR analyses showed significant increases in the expression of schizophrenia-associated genes in CPT2 knockdown compared to control zebrafish. Taken together, these data demonstrate that zebrafish are a useful model for studying the importance of beta-oxidation for early vertebrate development and brain function. This study also presents novel findings linking CPT2 deficiency to the regulation of schizophrenia and neurodegenerative disease-associated genes.

Hidden hearing loss in a Charcot-Marie-Tooth type 1A mouse model.

Hidden hearing loss (HHL), a recently described auditory neuropathy characterized by normal audiometric thresholds but reduced sound-evoked cochlear compound action potentials, has been proposed to contribute to hearing difficulty in noisy environments in people with normal hearing thresholds, a widespread complaint. While most studies on HHL pathogenesis have focused on inner hair cell (IHC) synaptopathy, we recently showed that transient auditory nerve (AN) demyelination also causes HHL in mice. To test the impact of myelinopathy on hearing in a clinically relevant model, we studied a mouse model of Charcot-Marie-Tooth type 1A (CMT1A), the most prevalent hereditary peripheral neuropathy in humans. CMT1A mice exhibit the functional hallmarks of HHL together with disorganization of AN heminodes near the IHCs with minor loss of AN fibers. These results support the hypothesis that mild disruptions of AN myelination can cause HHL, and that heminodal defects contribute to the alterations in the sound-evoked cochlear compound action potentials seen in this mouse model. Also, these findings suggest that patients with CMT1A or other mild peripheral neuropathies are likely to suffer from HHL. Furthermore, these results suggest that studies of hearing in CMT1A patients might help develop robust clinical tests for HHL, which are currently lacking.

Thin Ga2O3 Layers by Thermal Oxidation of van der Waals GaSe Nanostructures for Ultraviolet Photon Sensing.

Two-dimensional semiconductors (2DSEM) based on van der Waals crystals offer important avenues for nanotechnologies beyond the constraints of Moore's law and traditional semiconductors, such as silicon (Si). However, their application necessitates precise engineering of material properties and scalable manufacturing processes. The ability to oxidize Si to form silicon dioxide (SiO2) was crucial for the adoption of Si in modern technologies. Here, we report on the thermal oxidation of the 2DSEM gallium selenide (GaSe). The nanometer-thick layers are grown by molecular beam epitaxy on transparent sapphire (Al2O3) and feature a centro-symmetric polymorph of GaSe. Thermal annealing of the layers in an oxygen-rich environment promotes the chemical transformation and full conversion of GaSe into a thin layer of crystalline Ga2O3, paralleled by the formation of coherent Ga2O3/Al2O3 interfaces. Versatile functionalities are demonstrated in photon sensors based on GaSe and Ga2O3, ranging from electrical insulation to unfiltered deep ultraviolet optoelectronics, unlocking the technological potential of GaSe nanostructures and their amorphous and crystalline oxides.

Extent of Radiolytic Damage from Liquid Cell TEM Experiments on Metal-Organic Frameworks via Post-Mortem 4D-STEM.

We report a systematic analysis of electron beam damage of the zeolitic imidazolate framework (ZIF-8) during liquid cell transmission electron microscopy (LCTEM). Our analysis reveals ZIF-8 morphology is strongly affected by solvent used (water vs dimethylformamide), electron flux applied, and imaging mode (i.e., TEM vs STEM), while ZIF-8 crystallinity is primarily affected by accumulated electron fluence. Our observations indicate that the stability of ZIF-8 morphology is higher in dimethylformamide (DMF) than in water. However, in situ electron diffraction indicates that ZIF-8 nanocrystals lose crystallinity at critical fluence of ∼80 e-Å-2 independent of the presence of solvent. Furthermore, 4D-STEM analysis as a post-mortem method reveals the extent of electron beam damage beyond the imaging area and indicates that radiolytic reactions are more pronounced in TEM mode than in STEM mode. These results illustrate the significance of radiolysis occurring while imaging ZIF-8 and present a workflow for assessing damage in LCTEM experiments.

Conformational modulation and polymerization-induced folding of proteomimetic peptide brush polymers.

Peptide-brush polymers generated by graft-through living polymerization of peptide-modified monomers exhibit high proteolytic stability, therapeutic efficacy, and potential as functional tandem repeat protein mimetics. Prior work has focused on polymers generated from structurally disordered peptides that lack defined conformations. To obtain insight into how the structure of these polymers is influenced by the folding of their peptide sidechains, a set of polymers with varying degrees of polymerization was prepared from peptide monomers that adopt α-helical secondary structure for comparison to those having random coil structures. Circular dichroism and nuclear magnetic resonance spectroscopy confirm the maintenance of the secondary structure of the constituent peptide when polymerized. Small-angle X-ray scattering (SAXS) studies reveal the solution-phase conformation of PLPs in different solvent environments. In particular, X-ray scattering shows that modulation of solvent hydrophobicity, as well as hydrogen bonding patterns of the peptide sidechain, plays an important role in the degree of globularity and conformation of the overall polymer, with polymers of helical peptide brushes showing less spherical compaction in conditions where greater helicity is observed. These structural insights into peptide brush folding and polymer conformation inform the design of these proteomimetic materials with promise for controlling and predicting their artificial fold and morphology.

Associating transcription factors to single-cell trajectories with DREAMIT.

Inferring gene regulatory networks from single-cell RNA-sequencing trajectories has been an active area of research yet methods are still needed to identify regulators governing cell transitions. We developed DREAMIT (Dynamic Regulation of Expression Across Modules in Inferred Trajectories) to annotate transcription-factor activity along single-cell trajectory branches, using ensembles of relations to target genes. Using a benchmark representing several different tissues, as well as external validation with ATAC-Seq and Perturb-Seq data on hematopoietic cells, the method was found to have higher tissue-specific sensitivity and specificity over competing approaches.

Will there be water? Climate change, housing needs, and future water demand in California.

Climate change in California is expected to alter future water availability, impacting water supplies needed to support future housing growth and agriculture demand. In groundwater-dependent regions like California's Central Coast, new land-use related water demand and decreasing recharge is already stressing depleted groundwater basins. We developed a spatially explicit state-and-transition simulation model that integrates climate, land-use change, water demand, and groundwater gain-loss to examine the impact of future climate and land use change on groundwater balance and water demand in five counties along the Central Coast from 2010 to 2060. The model incorporated downscaled groundwater recharge projections based on a Warm/Wet and a Hot/Dry climate future from a spatially explicit hydrological process-based model. Two urbanization projections from a parcel-based, regional urban growth model representing 1) recent historical and 2) state-mandated housing growth projections were used as alternative spatial targets for future urban growth. Agricultural projections were based on recent historical trends from remote sensing data. Annual projected changes in groundwater balance were calculated as the difference between land-use related water demand, based on historical estimates, and climate-driven recharge plus agriculture return flows. Results indicate that future changes in climate-driven groundwater recharge, coupled with cumulative increases in agricultural water demand, result in overall declines in future groundwater balance, with a Hot/Dry future resulting in cumulative groundwater decline in all but Santa Cruz County. Cumulative declines by 2060 are especially prominent in San Luis Obispo (-2.9 to -5.1 Bm3) and Monterey counties (-6.5 to -8.7 Bm3), despite limited changes in agricultural water demand over the model period. These two counties show declining groundwater reserves in a Warm/Wet future as well, while San Benito and Santa Barbara County barely reach equilibrium. These results suggest future groundwater supplies may not be able to keep pace with regional demand and declining climate-driven recharge, resulting in a potential reduction in water security in the region. However, our county-scale projections showed new housing and associated water demand does not conflict with California's groundwater sustainability goals. Rather, future climate coupled with increasing agricultural groundwater demand may reduce water security in some counties, potentially limiting available groundwater supplies for new housing.

Molecular markers of proliferation, DNA repair, and immune infiltration defines high-risk subset of resectable retroperitoneal sarcomas.

INTRODUCTION: For retroperitoneal sarcomas (RPS), aggressive surgical resection offers the only chance for a cure; however, 5-year survival remains below 65%. Therefore, there is a critical need to identify drivers of poor clinical outcomes. MATERIALS AND METHODS: To identify biomarkers of tumors likely to recur following curative intent resection, we performed genomic and transcriptomic sequencing for 47 and 34 patients, respectively, with non-metastatic RPS at a single, high-volume sarcoma center. RESULTS: At the DNA level, alterations in TERT were associated with poor disease-free survival (DFS) and overall survival (OS). Increased RNA expression of gene sets related to growth signaling and DNA repair were associated with poor DFS and OS. Infiltration of CD8+ T-Cells and activated dendritic cells were associated with poor DFS and OS. CONCLUSION: These findings may help to better identify and treat non-metastatic, high-risk RPS.

The role of heterochronic gene expression and regulatory architecture in early developmental divergence.

New developmental programs can evolve through adaptive changes to gene expression. The annelid Streblospio benedicti has a developmental dimorphism, which provides a unique intraspecific framework for understanding the earliest genetic changes that take place during developmental divergence. Using comparative RNAseq through ontogeny, we find that only a small proportion of genes are differentially expressed at any time, despite major differences in larval development and life history. These genes shift expression profiles across morphs by either turning off any expression in one morph or changing the timing or amount of gene expression. We directly connect the contributions of these mechanisms to differences in developmental processes. We examine F1 offspring - using reciprocal crosses - to determine maternal mRNA inheritance and the regulatory architecture of gene expression. These results highlight the importance of both novel gene expression and heterochronic shifts in developmental evolution, as well as the trans-acting regulatory factors in initiating divergence.

Septic shock in the immunocompromised cancer patient: a narrative review.

Immunosuppressed patients, particularly those with cancer, represent a momentous and increasing portion of the population, especially as cancer incidence rises with population growth and aging. These patients are at a heightened risk of developing severe infections, including sepsis and septic shock, due to multiple immunologic defects such as neutropenia, lymphopenia, and T and B-cell impairment. The diverse and complex nature of these immunologic profiles, compounded by the concomitant use of immunosuppressive therapies (e.g., corticosteroids, cytotoxic drugs, and immunotherapy), superimposed by the breakage of natural protective barriers (e.g., mucosal damage, chronic indwelling catheters, and alterations of anatomical structures), increases the risk of various infections. These and other conditions that mimic sepsis pose substantial diagnostic and therapeutic challenges. Factors that elevate the risk of progression to septic shock in these patients include advanced age, pre-existing comorbidities, frailty, type of cancer, the severity of immunosuppression, hypoalbuminemia, hypophosphatemia, Gram-negative bacteremia, and type and timing of responses to initial treatment. The management of vulnerable cancer patients with sepsis or septic shock varies due to biased clinical practices that may result in delayed access to intensive care and worse outcomes. While septic shock is typically associated with poor outcomes in patients with malignancies, survival has significantly improved over time. Therefore, understanding and addressing the unique needs of cancer patients through a new paradigm, which includes the integration of innovative technologies into our healthcare system (e.g., wireless technologies, medical informatics, precision medicine), targeted management strategies, and robust clinical practices, including early identification and diagnosis, coupled with prompt admission to high-level care facilities that promote a multidisciplinary approach, is crucial for improving their prognosis and overall survival rates.