Peptide programming of supramolecular vinylidene fluoride ferroelectric phases.

Ferroelectric structures have spontaneous macroscopic polarization that can be inverted using external electric fields and have potential applications including information storage, energy transduction, ultralow-power nanoelectronics1,2 and biomedical devices3. These functions would benefit from nanoscale control of ferroelectric structure, the ability to switch polarization with lower applied fields (low coercive field) and biocompatibility. Soft ferroelectrics based on poly(vinylidene fluoride) (PVDF)4-6 have a thermodynamically unstable ferroelectric phase in the homopolymer, complex semi-crystalline structures, and high coercive fields. Here we report on ferroelectric materials formed by water-soluble molecules containing only six VDF repeating units covalently conjugated to a tetrapeptide, with the propensity to assemble into the ß-sheet structures that are ubiquitous in proteins. This led to the discovery of ribbon-shaped ferroelectric supramolecular assemblies that are thermodynamically stable with their long axes parallel to both the preferred hydrogen-bonding direction of ß-sheets and the bistable polar axes of VDF hexamers. Relative to a commonly used ferroelectric copolymer, the biomolecular assemblies exhibit a coercive field that is two orders of magnitude lower, as the result of supramolecular dynamics, and a similar level of remnant polarization, despite having a peptide content of 49 wt%. Furthermore, the Curie temperature of the assemblies is about 40 °C higher than that of a copolymer containing a similar amount of VDF. This supramolecular system was created using a biologically inspired strategy that is attractive in terms of sustainability and that could lead to new functions for soft ferroelectrics.

Variation in season length and development time is sufficient to drive the emergence and coexistence of social and solitary behavioural strategies.

Season length and its associated variables can influence the expression of social behaviours, including the occurrence of eusociality in insects. Eusociality can vary widely across environmental gradients, both within and between different species. Numerous theoretical models have been developed to examine the life history traits that underlie the emergence and maintenance of eusociality, yet the impact of seasonality on this process is largely uncharacterized. Here, we present a theoretical model that incorporates season length and offspring development time into a single, individual-focused model to examine how these factors can shape the costs and benefits of social living. We find that longer season lengths and faster brood development times are sufficient to favour the emergence and maintenance of a social strategy, while shorter seasons favour a solitary one. We also identify a range of season lengths where social and solitary strategies can coexist. Moreover, our theoretical predictions are well matched to the natural history and behaviour of two flexibly eusocial bee species, suggesting that our model can make realistic predictions about the evolution of different social strategies. Broadly, this work reveals the crucial role that environmental conditions can have in shaping social behaviour and its evolution and it underscores the need for further models that explicitly incorporate such variation to study the evolutionary trajectories of eusociality.

Integer Factorization with Stochastic Spiking in Percolating Networks of Nanoparticles.

As growth in global demand for computing power continues to outpace ongoing improvements in transistor-based hardware, novel computing solutions are required. One promising approach employs stochastic nanoscale devices to accelerate probabilistic computing algorithms. Percolating Networks of Nanoparticles (PNNs) exhibit stochastic spiking, which is of particular interest as it meets criteria for criticality which is associated with a range of computational advantages. Here, we show several ways in which spiking PNNs can be used as the core stochastic components of coupled networks that allow successful factorization of integers up to 945. We demonstrate asynchronous operation and show that a single device is sufficient to solve all factorization tasks and to generate multiple solutions simultaneously.

A cautious approach to subsidies for environmental sustainability.

Transformational change is possible, but design and implementation must seek to avoid lock-in.

Multiomic single-cell sequencing defines tissue-specific responses in Stevens-Johnson syndrome and toxic epidermal necrolysis.

Stevens-Johnson syndrome and toxic epidermal necrolysis (SJS/TEN) is a rare but life-threatening cutaneous drug reaction mediated by human leukocyte antigen (HLA) class I-restricted CD8+ T cells. For unbiased assessment of cellular immunopathogenesis, here we perform single-cell (sc) transcriptome, surface proteome, and T cell receptor (TCR) sequencing on unaffected skin, affected skin, and blister fluid from 15 SJS/TEN patients. From 109,888 cells, we identify 15 scRNA-defined subsets. Keratinocytes express markers indicating HLA class I-restricted antigen presentation and appear to trigger the proliferation of and killing by cytotoxic CD8+ tissue-resident T cells that express granulysin, granzyme B, perforin, LAG3, CD27, and LINC01871, and signal through the PKM, MIF, TGFß, and JAK-STAT pathways. In affected tissue, cytotoxic CD8+ T cells express private expanded and unexpanded TCRαß that are absent or unexpanded in unaffected skin, and mixed populations of macrophages and fibroblasts express pro-inflammatory markers or those favoring repair. This data identifies putative cytotoxic TCRs and therapeutic targets.

Utility of combining frailty and comorbid disease indices in predicting outcomes following craniotomy for adult primary brain tumors: A mixed-effects model analysis using the nationwide readmissions database.

OBJECTIVE: The escalating healthcare expenditures in the United States, particularly in neurosurgery, necessitate effective tools for predicting patient outcomes and optimizing resource allocation. This study explores the utility of combining frailty and comorbidity indices, specifically the Johns Hopkins Adjusted Clinical Groups (JHACG) frailty index and the Elixhauser Comorbidity Index (ECI), in predicting hospital length of stay (LOS), non-routine discharge, and one-year readmission in patients undergoing craniotomy for benign and malignant primary brain tumors. METHODS: Leveraging the Nationwide Readmissions Database (NRD) for 2016-2019, we analyzed data from 645 patients with benign and 30,991 with malignant tumors. Frailty, ECI, and frailty + ECI were assessed as predictors using generalized linear mixed-effects models. Receiver operating characteristic (ROC) curves evaluated predictive performance. RESULTS: Patients in the benign tumor cohort had a mean LOS of 8.1 ± 15.1 days, and frailty + ECI outperformed frailty alone in predicting non-routine discharge (AUC 0.829 vs. 0.820, p = 0.035). The malignant tumor cohort patients had a mean LOS of 7.9 ± 9.1 days. In this cohort, frailty + ECI (AUC 0.821) outperformed both frailty (AUC 0.744, p < 0.0001) and ECI alone (AUC 0.809, p < 0.0001) in predicting hospital LOS. Frailty + ECI (AUC 0.831) also proved superior to frailty (AUC 0.809, p < 0.0001) and ECI alone (AUC 0.827, p < 0.0001) in predicting non-routine discharge location for patients with malignant tumors. All indices performed comparably to one another as a predictor of readmission in both cohorts. CONCLUSION: This study highlights the synergistic predictive capacity of frailty + ECI, especially in malignant tumor cases, and further suggests that comorbid diseases may greatly influence perioperative outcomes more than frailty. Enhanced risk assessment could aid clinical decision-making, patient counseling, and resource allocation, ultimately optimizing patient outcomes.

A rapid non-invasive wearable device for assessing cardiac troponin.

BACKGROUND: Chest pain, a common emergency department 35 (ED) presentation, requires rapid evaluation. Optical technology-based non-invasive wearable devices (Infrasensor, RCE, Carlsbad, CA) rapidly and transcutaneously assesses cardiac Troponin I (cTnI). OBJECTIVES: To perform a pilot study describing the performance of the Infrasensor in cTnI defined cohorts. METHODS: This was a 10-hospital prospective observational study in healthy US subjects with a normal cTnI, and in patients with an elevated local cTnI. Healthy subjects were without disease, defined by a negative questionnaire and bloodwork, had a 3-minute Infrasensor measurement and blood samples for high-sensitivity cardiac troponin I (hs-cTnI), n-terminal pro-B-type natriuretic peptide (NTproBNP), creatinine, and glycosylated hemoglobin (HbA1c). Elevated cTnI's patients had the same Infrasensor and blood sample measurements. Using a cross validation technique, a cTnI based binary classification model that did, and did not, include age was trained with 80%, and validated on 20% (n=168; elevated hs-cTnI equally distributed across 5 folds) of the overall cohort. RESULTS: Of 840 patients, 727 (87.5%) were non-elevated cTnI controls and the remainder, n=113, had elevated cTnI. Median (25th, 75th percentiles) age was 61 (52, 71) and 48 (32, 57) years for the elevated and healthy control cohorts, respectively. Overall, 50.5% were female, with 29.2% and 52.7% in the elevated and non-elevated troponin cohorts respectively. Overall, the sensitivity, specificity, negative and positive predictive values of the Infrasensor for identifying an elevated cTnI were 0.9, 0.7, 0.98 and 0.48 respectively, with a C-statistic of 0.90 (0.89-0.99). CONCLUSIONS: The Infrasensor identifies elevated cTnI within 3 minutes of application.

Long-term disability trajectories in multiple sclerosis: a group-based trajectory analysis of the AusLong cohort.

BACKGROUND: Previous natural history studies highlighted a consistent heterogeneity of disability trajectories among individuals with primary or secondary progressive multiple sclerosis (MS). However, evidence on disability progression in relapsing onset MS is scarce.The aim of this study was to investigate heterogeneity in disability accumulation over 10 years following a first clinical diagnosis of central nervous system demyelination (FCD) and identify genetic, demographic, environmental and clinical factors associated with these trajectories. METHODS: We used group-based trajectory models to measure heterogeneity in disability trajectories based on the Expanded Disability Status Scale (EDSS) in a prospectively assessed cohort of 263 participants. To capture sustained neurological impairments and avoid issues related to significant changes in EDSS associated with relapse, we did not consider EDSS points recorded within 3 months of a relapse. RESULTS: We identified three distinct and clinically meaningful disability trajectories: No/minimal, moderate and severe. Those in the no/minimal disability trajectory showed no appreciable progression of disability (median EDSS∼1 at 10-year review) while those in the moderate and severe disability trajectories experienced disability worsening (median time to reach EDSS 4 was 9 and 7 years, respectively). Compared with the no/minimal disability trajectory, those with older age, a higher number of relapses within the first 5 years post-FCD, and a higher number of comorbidities at baseline were more likely to be in the worse disability trajectory. Surprisingly, baseline MRI and anatomical site of initial symptoms did not influence long-term outcomes. CONCLUSIONS: Those at higher risk of faster MS disability progression can be identified based on their early clinical characteristics with potential therapeutic implications for early intervention and treatment escalation.

Evaluation of early treatment with intravenous idursulfase and intrathecal idursulfase-IT on cognitive function in siblings with neuronopathic mucopolysaccharidosis II.

Mucopolysaccharidosis II (MPS II; Hunter syndrome; OMIM 309900) is a rare, X-linked, heterogeneous lysosomal storage disease. Approximately two-thirds of patients develop cognitive impairment, which is difficult to assess in clinical trials, partly owing to the variable nature of cognitive impairment. Analyzing data from siblings can help to minimize this heterogeneity. We report analyses of cognitive function from siblings with MPS II enrolled in clinical trials: a natural history study (NCT01822184), a randomized, open-label, phase 2/3 study of intravenous (IV) idursulfase with or without intrathecal idursulfase (idursulfase-IT; NCT02055118), and its extension (NCT2412787). Cognitive function was assessed using Differential Abilities Scales, Second Edition General Conceptual Ability (DAS-II GCA) scores; Bayley Scales of Infant and Toddler Development, Third Edition; and Vineland Adaptive Behavior Scales, Second Edition Adaptive Behavior Composite (VABS-II ABC). Seven sets of siblings (six pairs and one set of three) were included. All patients received IV idursulfase and 10 received subsequent idursulfase-IT. Younger siblings initiated IV idursulfase at an earlier age than their older sibling(s) in six of the sets; the younger sibling started treatment before 1 year of age in three sets. Monthly idursulfase-IT was generally associated with a stabilization of cognitive function: DAS-II GCA and VABS-II ABC scores were higher at age-matched assessments in the majority of those who either received idursulfase-IT earlier than their sibling or who received idursulfase-IT versus no idursulfase-IT. These data suggest that early initiation of intrathecal enzyme replacement therapy may stabilize or slow cognitive decline in some patients with neuronopathic MPS II.

Biallelic PTPMT1 variants disrupt cardiolipin metabolism and lead to a neurodevelopmental syndrome.

Primary mitochondrial diseases (PMDs) are among the most common inherited neurological disorders. They are caused by pathogenic variants in mitochondrial or nuclear DNA that disrupt mitochondrial structure and/or function, leading to impaired oxidative phosphorylation (OXPHOS). One emerging subcategory of PMDs involves defective phospholipid (PL) metabolism. Cardiolipin (CL), the signature PL of mitochondria, resides primarily in the inner mitochondrial membrane, where it is biosynthesised and remodelled via multiple enzymes and is fundamental to several aspects of mitochondrial biology. Genes that contribute to CL biosynthesis have recently been linked with PMD. However, the pathophysiological mechanisms that underpin human CL-related PMDs are not fully characterised. Here, we report six individuals, from three independent families, harbouring biallelic variants in PTPMT1, a mitochondrial tyrosine phosphatase required for de novo CL biosynthesis. All patients presented with a complex, neonatal/infantile onset neurological and neurodevelopmental syndrome comprising developmental delay, microcephaly, facial dysmorphism, epilepsy, spasticity, cerebellar ataxia and nystagmus, sensorineural hearing loss, optic atrophy, and bulbar dysfunction. Brain MRI revealed a variable combination of corpus callosum thinning, cerebellar atrophy, and white matter changes. Using patient-derived fibroblasts and skeletal muscle tissue, combined with cellular rescue experiments, we characterise the molecular defects associated with mutant PTPMT1 and confirm the downstream pathogenic effects that loss of PTPMT1 has on mitochondrial structure and function. To further characterise the functional role of PTPMT1 in CL homeostasis, we established a zebrafish ptpmt1 knockout model associated with abnormalities in body size, developmental alterations, decreased total CL levels, and OXPHOS deficiency. Together, these data indicate that loss of PTPMT1 function is associated with a new autosomal recessive PMD caused by impaired CL metabolism, highlight the contribution of aberrant CL metabolism towards human disease, and emphasise the importance of normal CL homeostasis during neurodevelopment.

Catalytic Heaters at Oil and Gas Sites May be a Significant yet Overlooked Seasonal Source of Methane Emissions.

Successful reduction of oil and gas sector methane emissions to meet near-zero intensity targets requires the identification and mitigation of all possible sources. One potentially important source is catalytic heaters, which have largely escaped attention in regulatory and mitigation efforts despite being ubiquitous at upstream production sites in cold climate regions. This study reports direct in situ measurements of the exhaust streams of 38 natural gas-fired catalytic heaters at upstream production sites in British Columbia, Canada. All heaters in the sample showed consistently poor methane conversion with mean destruction efficiencies of 61 ± 5% while releasing 235 [+31/-28] g of methane per cubic meter of fuel. Although individual units are generally small methane sources (mean of 0.28 ± 0.04 kg/h), their prevalence means they could represent 6% of the total provincial upstream methane inventory and as an aggregate methane source could be 5× more significant than abandoned wells. Notably, these heaters are seasonal sources whose emissions would be missed in measurement campaigns occurring solely in summer months. However, additional measurements from a small number of heat medium burners demonstrate that, where feasible, methane emissions can be reduced by approximately 425× by replacing catalytic heaters with centralized heat systems.

Guiding Principles for Department of Defense Global Health Engagements.

Within the past decade, the U.S. DoD has increased spending on global health engagements (GHEs). Not only do these increased efforts strengthen national security but they also fortify the United States' diplomatic relations with foreign governments through medical stability operations and health diplomacy. Maximizing health care-related programs will bolster a partner government's ability to maintain secure borders and prevent conflict while depriving extremists of safe havens. Looking beyond the short-term benefits, the long-term effects of GHEs are not limited to a nation's borders; aid permeates into a society providing social and economic opportunities otherwise unobtainable to the community and effectively winning the hearts and minds of the population. Although the DoD does list policies and procedures in which to conduct GHEs, the proposed set of principles can further prevent conflicts, deliver better foreign humanitarian assistance, and develop a better health care partnership centered around the host nation.

Advance directives in the intensive care unit: An eight-year vanguard cohort study.

PURPOSE: To investigate the frequency, content, and clinical translation of advance directives in intensive care units (ICUs). MATERIAL AND METHODS: Retrospective cohort study in a Swiss tertiary ICU, including patients with advance directives treated in ICUs ≥48 h. The primary endpoint was the violation of directives. Key secondary endpoints were the directives' prevalence and their translation into clinical practice. RESULTS: Of 5'851 patients treated ≥48 h in ICUs, 2.7 % had documented directives. Despite 92 % using templates, subjective or contradictory wording was found in 19 % and 12 %. Nine percent of directives were violated. Patients with directive violations had worse in-hospital outcomes (p = 0.012). At admission, 64 % of patients experiencing violations could not communicate, and directives were missing/unrecognized in 30 %. Mostly, directives were not followed regarding life-prolonging measures (6 %), ICU admission (5 %), and mechanical ventilation (3 %). Kaplan Meier statistics revealed a lower survival rate with directives recognized at admission (p = 0.04) and when treatment was withheld (p < 0.001). CONCLUSIONS: Advance directives are available in a minority of ICU patients and often contain subjective/contradictory wording. Physicians respected directives in 90 % of patients, with treatment adapted following their wishes. However, violation of directives may have serious consequences with unfavorable in-hospital outcomes and decreased long-term survival with treatment adaption following directives.

Improving on polygenic scores across complex traits using select and shrink with summary statistics (S4) and LDpred2.

BACKGROUND: As precision medicine advances, polygenic scores (PGS) have become increasingly important for clinical risk assessment. Many methods have been developed to create polygenic models with increased accuracy for risk prediction. Our select and shrink with summary statistics (S4) PGS method has previously been shown to accurately predict the polygenic risk of epithelial ovarian cancer. Here, we applied S4 PGS to 12 phenotypes for UK Biobank participants, and compared it with the LDpred2 and a combined S4 + LDpred2 method. RESULTS: The S4 + LDpred2 method provided overall improved PGS accuracy across a variety of phenotypes for UK Biobank participants. Additionally, the S4 + LDpred2 method had the best estimated PGS accuracy in Finnish and Japanese populations. We also addressed the challenge of limited genotype level data by developing the PGS models using only GWAS summary statistics. CONCLUSIONS: Taken together, the S4 + LDpred2 method represents an improvement in overall PGS accuracy across multiple phenotypes and populations.

Advancing age grading techniques for Glossina morsitans morsitans, vectors of African trypanosomiasis, through mid-infrared spectroscopy and machine learning.

Tsetse are the insects responsible for transmitting African trypanosomes, which cause sleeping sickness in humans and animal trypanosomiasis in wildlife and livestock. Knowing the age of these flies is important when assessing the effectiveness of vector control programs and modelling disease risk. Current methods to assess fly age are, however, labour-intensive, slow, and often inaccurate as skilled personnel are in short supply. Mid-infrared spectroscopy (MIRS), a fast and cost-effective tool to accurately estimate several biological traits of insects, offers a promising alternative. This is achieved by characterising the biochemical composition of the insect cuticle using infrared light coupled with machine-learning (ML) algorithms to estimate the traits of interest. We tested the performance of MIRS in estimating tsetse sex and age for the first-time using spectra obtained from their cuticle. We used 541 insectary-reared Glossina m. morsitans of two different age groups for males (5 and 7 weeks) and three age groups for females (3 days, 5 weeks, and 7 weeks). Spectra were collected from the head, thorax, and abdomen of each sample. ML models differentiated between male and female flies with a 96% accuracy and predicted the age group with 94% and 87% accuracy for males and females, respectively. The key infrared regions important for discriminating sex and age classification were characteristic of lipid and protein content. Our results support the use of MIRS as a rapid and accurate way to identify tsetse sex and age with minimal pre-processing. Further validation using wild-caught tsetse could pave the way for this technique to be implemented as a routine surveillance tool in vector control programmes.

Sex-specific high-sensitivity troponin T cut-points have similar safety but lower efficacy than overall cut-points in a multisite U.S. cohort.

BACKGROUND: Data comparing the performance of sex-specific to overall (non-sex-specific) high-sensitivity cardiac troponin (hs-cTn) cut-points for diagnosing acute coronary syndrome (ACS) are limited. This study aims to compare the safety and efficacy of sex-specific versus overall 99th percentile high-sensitivity cardiac troponin T (hs-cTnT) cut-points. METHODS: We conducted a secondary analysis of the STOP-CP cohort, which prospectively enrolled emergency department patients ≥ 21 years old with symptoms suggestive of ACS without ST-elevation on initial electrocardiogram across eight U.S. sites (January 25, 2017-September 6, 2018). Participants with both 0- and 1-h hs-cTnT measures less than or equal to the 99th percentile (sex-specific 22 ng/L for males, 14 ng/L for females; overall 19 ng/L) were classified into the rule-out group. The safety outcome was adjudicated cardiac death or myocardial infarction (MI) at 30 days. Efficacy was defined as the proportion classified to the rule-out group. McNemar's test and a generalized score statistic were used to compare rule-out and 30-day cardiac death or MI rates between strategies. Net reclassification improvement (NRI) index was used to further compare performance. RESULTS: This analysis included 1430 patients, of whom 45.8% (655/1430) were female; the mean ± SD age was 57.6 ± 12.8 years. At 30 days, cardiac death or MI occurred in 12.8% (183/1430). The rule-out rate was lower using sex-specific versus overall cut-points (70.6% [1010/1430] vs. 72.5% [1037/1430]; p = 0.003). Among rule-out patients, the 30-day cardiac death or MI rates were similar for sex-specific (2.4% [24/1010]) vs. overall (2.3% [24/1037]) strategies (p = 0.79). Among patients with cardiac death or MI, sex-specific versus overall cut-points correctly reclassified three females and incorrectly reclassified three males. The sex-specific strategy resulted in a net of 27 patients being incorrectly reclassified into the rule-in group. This led to an NRI of -2.2% (95% CI -5.1% to 0.8%). CONCLUSIONS: Sex-specific hs-cTnT cut-points resulted in fewer patients being ruled out without an improvement in safety compared to the overall cut-point strategy.

Delineation of the impact on temporal behaviors of off-axis photoemission in an ultrafast electron microscope.

Efforts to push the spatiotemporal imaging-resolution limits of femtosecond laser-driven ultrafast electron microscopes (UEMs) to the combined angstrom-fs range will benefit from stable sources capable of generating high bunch charges. Recent demonstrations of unconventional off-axis photoemitting geometries are promising, but connections to the observed onset of structural dynamics are yet to be established. Here we use the in-situ photoexcitation of coherent phonons to quantify the relative time-of-flight (r-TOF) of photoelectron packets generated from the Ni Wehnelt aperture and from a Ta cathode set-back from the aperture plane. We further support the UEM experiments with particle-tracing simulations of the precise electron-gun architecture and photoemitting geometries. In this way, we measure discernible shifts in electron-packet TOF of tens of picoseconds for the two photoemitting surfaces. These shifts arise from the impact that the Wehnelt-aperture off-axis orientation has on the electron-momentum distribution, which modifies both the collection efficiency and the temporal-packet distribution relative to on-axis emission. Future needs are identified; we expect this and other developments in UEM electron-gun configuration to expand the range of material phenomena that can be directly imaged on scales commensurate with fundamental structural dynamics.

Cytokine and chemokine receptor profiles in adipose tissue vasculature unravel endothelial cell responses in HIV.

Chronic systemic inflammation significantly increases myocardial infarction risk in people living with HIV (PLWH). Endothelial cell dysfunction disrupts vascular homeostasis regulation, increasing the risk of vasoconstriction, inflammation, and thrombosis, contributing to cardiovascular disease. We aimed to characterize endothelial cell (EC) chemokines, cytokine, and chemokine receptors of PLWH, hypothesizing that in our cohort, glucose intolerance contributes to their differential expression implicated in endothelial dysfunction. Using single-cell transcriptomic analysis, we phenotyped chemokine and cytokine receptor expression on arterial ECs, capillary ECs, venous ECs, and vascular smooth muscle cells (VSMCs) in subcutaneous adipose tissue of 59 PLWH with and without glucose intolerance. Our results show that arterial and capillary ECs express significantly higher interferon and tumor necrosis factor (TNF) receptors than venous ECs and VSMCs. Venous ECs exhibited more interleukin (IL)1R1 and ACKR1 receptors, and VSMCs showed significant IL6R expression than arterial and capillary ECs. When stratified by group, arterial ECs from PLWH with glucose intolerance expressed significantly higher IL1R1, IL6R, CXCL12, CCL14, and ICAM2 transcripts than arterial ECs from PLWH without diabetes. Of the different vascular cell types studied, arterial ECs as a proportion of all ECs in adipose tissue were positively correlated with plasma fasting blood glucose. In contrast, venous ECs and VSMCs were positively correlated with plasma IL6. To directly assess the effect of plasma from PLWH on endothelial function, we cultured human arterial ECs (HAECs) in plasma-conditioned media from PLWH and performed bulk RNA sequencing. Plasma from PLWH stimulated ECs with the upregulation of genes that enrich for the oxidative phosphorylation and the TNF-α via NFK-ß pathways. In conclusion, ECs in PLWH show heterogeneous cytokine and chemokine receptor expression, and arterial ECs were the most influenced by glucose intolerance. Further research must explicate cytokine and chemokine roles in EC dysfunction and identify biomarkers for disease progression and therapeutic response.

Repurposing an Endogenous CRISPR-Cas System to Generate and Study Subtle Mutations in Bacteriophages.

While bacteriophage applications benefit from effective phage engineering, selecting the desired genotype after subtle modifications remains challenging. Here, we describe a two-phase endogenous CRISPR-Cas-based phage engineering approach that enables selection of small defined edits in Pectobacterium carotovorum phage ZF40. We designed plasmids containing sequences homologous to ZF40 and a mini-CRISPR array. The plasmids allowed genome editing through homologous recombination and counter-selection against non-recombinant phage genomes using an endogenous type I-E CRISPR-Cas system. With this technique, we first deleted target genes and subsequently restored loci with modifications. This two-phase approach circumvented major challenges in subtle phage modifications, including inadequate sequence distinction for CRISPR-Cas counter-selection and the requirement of a protospacer-adjacent motif, limiting sequences that can be modified. Distinct 20-bp barcodes were incorporated through engineering as differential target sites for programmed CRISPR-Cas activity, which allowed quantification of phage variants in mixed populations. This method aids studies and applications that require mixtures of similar phages.

Streamlining LC-MS Characterization of Pharmaceutical Polymers by Fourier-Transform-Based Deconvolution and Macromolecular Mass Defect Analysis.

Polymer conjugation has risen in importance over the past three decades as a means of increasing the in vivo half-life of biotherapeutics, with benefits including better stability, greater drug efficacy, and lower toxicity. However, the intrinsic variability of polymer synthesis results in products with broad distributions in chain length and branching structure, complicating quality control for successful functionalization and downstream conjugation. Frequently, a combination of several analytical techniques is required for comprehensive characterization. While liquid chromatography-mass spectrometry (LC-MS) is a powerful platform that can provide detailed molecular features of polymers, the mass spectra are inherently challenging to interpret due to high mass polydispersity and overlapping charge distributions. Here, by leveraging Fourier transform-based deconvolution and macromolecular mass defect analysis, we demonstrate a new way to streamline pharmaceutical polymer analysis, shedding light on polymer size, composition, branching, and end-group functionalization with the capability for reaction monitoring.