An IgM Cleaving Enzyme for Clearance of Anti-Pig Xenoreactive Antibodies in a Nonhuman Primate Model.

Background: The removal of preformed antibodies with cleaving enzyme like IdeS (Imlifidase) has demonstrated therapeutic potential in organ transplantation for sensitized recipients. However, preformed xenoreactive antibodies (XAbs) against porcine glycans are predominantly IgM and considered detrimental in pig-to-human xenotransplantation.

Methods: Recombinant IceM, an endopeptidase cleaving IgM, was generated in Escherichia coli.

Reduced time spent with patients and decreased satisfaction in work during COVID-19 pandemic.

Background: The COVID-19 pandemic disrupted the healthcare system, affecting physician wellbeing. The consequences of reduced time spent with patients at bedside during the pandemic has not been investigated. The objectives of this study include assessing time spent with patients, physician wellbeing and patient satisfaction before and during the pandemic.

Engineered IgM and IgG cleaving enzymes for mitigating antibody neutralization and complement activation in AAV gene transfer.

Systemic dosing of adeno-associated viral (AAV) vectors poses potential risk of adverse side effects including complement activation triggered by anti-capsid immunity. Due to the multifactorial nature of toxicities observed in this setting, a wide spectrum of immune modulatory regimens are being investigated in the clinic. Here, we discover an IgM cleaving enzyme (IceM) that degrades human IgM, a key trigger in the anti-AAV immune cascade.

Capsid-mediated control of adeno-associated viral transcription determines host range.

Adeno-associated virus (AAV) is a member of the genus Dependoparvovirus, which infects a wide range of vertebrate species. Here, we observe that, unlike most primate AAV isolates, avian AAV is transcriptionally silenced in human cells. By swapping the VP1 N terminus from primate AAVs (e.

Interplay between Furin and Sialoglycans in Modulating Adeno-Associated Viral Cell Entry.

Adeno-associated viruses (AAVs) are small, helper-dependent, single-stranded DNA viruses that exploit a broad spectrum of host factors for cell entry. During the course of infection, several AAV serotypes have been shown to transit through the -Golgi network within the host cell. In the current study, we investigated whether the Golgi-localized, calcium-dependent protease furin influences AAV transduction.

An organ-on-chip model of pulmonary arterial hypertension identifies a BMPR2-SOX17-prostacyclin signalling axis.

Pulmonary arterial hypertension (PAH) is an unmet clinical need. The lack of models of human disease is a key obstacle to drug development. We present a biomimetic model of pulmonary arterial endothelial-smooth muscle cell interactions in PAH, combining natural and induced bone morphogenetic protein receptor 2 (BMPR2) dysfunction with hypoxia to induce smooth muscle activation and proliferation, which is responsive to drug treatment.

(Phenyl-iodos-yl)benzene tosyl-ate dihydrate.

The structure of the title salt (systematic name: oxodiphenyl-λ-iodanylium 4-methyl-benzene-sulfonate dihydrate), CHIO·CHOS·2HO, at 150 K, has monoclinic (2/) symmetry. The mol-ecular structure features an angular (phenyl-iodos-yl)benzene cation, the geometry of which was hitherto undescribed in the literature: in the cation, both I-C bonds are approximately normal to the I=O bond, forming a C-I-C angle of 95.36 (4)°.

Cross-species evolution of a highly potent AAV variant for therapeutic gene transfer and genome editing.

Recombinant adeno-associated viral (AAV) vectors are a promising gene delivery platform, but ongoing clinical trials continue to highlight a relatively narrow therapeutic window. Effective clinical translation is confounded, at least in part, by differences in AAV biology across animal species. Here, we tackle this challenge by sequentially evolving AAV capsid libraries in mice, pigs and macaques.

Gigaxonin glycosylation regulates intermediate filament turnover and may impact giant axonal neuropathy etiology or treatment.

Gigaxonin (also known as KLHL16) is an E3 ligase adaptor protein that promotes the ubiquitination and degradation of intermediate filament (IF) proteins. Mutations in human gigaxonin cause the fatal neurodegenerative disease giant axonal neuropathy (GAN), in which IF proteins accumulate and aggregate in axons throughout the nervous system, impairing neuronal function and viability. Despite this pathophysiological significance, the upstream regulation and downstream effects of normal and aberrant gigaxonin function remain incompletely understood.

A Novel Glycoproteomics Workflow Reveals Dynamic O-GlcNAcylation of COPγ1 as a Candidate Regulator of Protein Trafficking.

O-linked β-N-acetylglucosamine (O-GlcNAc) is an abundant and essential intracellular form of protein glycosylation in animals and plants. In humans, dysregulation of O-GlcNAcylation occurs in a wide range of diseases, including cancer, diabetes, and neurodegeneration. Since its discovery more than 30 years ago, great strides have been made in understanding central aspects of O-GlcNAc signaling, including identifying thousands of its substrates and characterizing the enzymes that govern it.

Structural basis of O-GlcNAc recognition by mammalian 14-3-3 proteins.

O-GlcNAc is an intracellular posttranslational modification that governs myriad cell biological processes and is dysregulated in human diseases. Despite this broad pathophysiological significance, the biochemical effects of most O-GlcNAcylation events remain uncharacterized. One prevalent hypothesis is that O-GlcNAc moieties may be recognized by "reader" proteins to effect downstream signaling.

Site-specific glycosylation regulates the form and function of the intermediate filament cytoskeleton.

Intermediate filaments (IF) are a major component of the metazoan cytoskeleton and are essential for normal cell morphology, motility, and signal transduction. Dysregulation of IFs causes a wide range of human diseases, including skin disorders, cardiomyopathies, lipodystrophy, and neuropathy. Despite this pathophysiological significance, how cells regulate IF structure, dynamics, and function remains poorly understood.

Dynamic Glycosylation Governs the Vertebrate COPII Protein Trafficking Pathway.

The COPII coat complex, which mediates secretory cargo trafficking from the endoplasmic reticulum, is a key control point for subcellular protein targeting. Because misdirected proteins cannot function, protein sorting by COPII is critical for establishing and maintaining normal cell and tissue homeostasis. Indeed, mutations in COPII genes cause a range of human pathologies, including cranio-lenticulo-sutural dysplasia (CLSD), which is characterized by collagen trafficking defects, craniofacial abnormalities, and skeletal dysmorphology.

Esterified Trehalose Analogues Protect Mammalian Cells from Heat Shock.

Trehalose is a disaccharide produced by many organisms to better enable them to survive environmental stresses, including heat, cold, desiccation, and reactive oxygen species. Mammalian cells do not naturally biosynthesize trehalose; however, when introduced into mammalian cells, trehalose provides protection from damage associated with freezing and drying. One of the major difficulties in using trehalose as a cellular protectant for mammalian cells is the delivery of this disaccharide into the intracellular environment; mammalian cell membranes are impermeable to the hydrophilic sugar trehalose.

Glycosylation of KEAP1 links nutrient sensing to redox stress signaling.

O-GlcNAcylation is an essential, nutrient-sensitive post-translational modification, but its biochemical and phenotypic effects remain incompletely understood. To address this question, we investigated the global transcriptional response to perturbations in O-GlcNAcylation. Unexpectedly, many transcriptional effects of O-GlcNAc transferase (OGT) inhibition were due to the activation of NRF2, the master regulator of redox stress tolerance.

Redox-triggered mixing and demixing of surfactants within assemblies formed in solution and at surfaces.

We report experiments that test the hypothesis that redox-triggered changes in the architectures of surfactants permit control of mixing of surfactants within assemblies. Specifically, we describe surface tension, light scattering, atomic force microscopy, and quartz crystal microbalance measurements that characterize the redox-dependent behaviors of cationic surfactants with a ferrocene group located either at the surfactant terminus (11-ferrocenylundecyl-trimethylammonium bromide; FTMA) or head (N,N-dimethylferrocenylmethyldecylammonium bromide; DMFA). In bulk solution, we find that reduced and oxidized FTMA do not mix within micellar assemblies but that reduced and oxidized DMFA do form mixed micelles.

HPV-associated oropharyngeal squamous cell carcinoma.

Human papillomavirus (HPV) can infect the tonsillar tissues of the oropharynx and is associated with oropharyngeal squamous cell carcinoma. This article provides an overview to guide primary care providers in screening patients for oropharyngeal cancer and making appropriate referrals. The article also reviews available HPV vaccines and immunization adherence rates.

A six-gene phylogeny provides new insights into choanoflagellate evolution.

Recent studies have shown that molecular phylogenies of the choanoflagellates (Class Choanoflagellatea) are in disagreement with their traditional taxonomy, based on morphology, and that Choanoflagellatea requires considerable taxonomic revision. Furthermore, phylogenies suggest that the morphological and ecological evolution of the group is more complex than has previously been recognized. Here we address the taxonomy of the major choanoflagellate order Craspedida, by erecting four new genera.

Influence of self-assembling redox mediators on charge transfer at hydrophobic electrodes.

We report an investigation of the influence of reversible self-assembly of amphiphilic redox-mediators on interfacial charge transfer at chemically functionalized electrodes. Specifically, we employed (11-ferrocenylundecyl)-trimethylammonium bromide (FTMA) as a model self-assembling redox mediator and alkanethiol-modified gold films as hydrophobic electrodes. By performing cyclic voltammetry (CV, 10 mV/s) in aqueous solutions containing FTMA above its critical micellar concentration (CMC), we measured anodic (Ia) and cathodic (Ic) peak current densities of 18 ± 3 and 1.

A chemical glycoproteomics platform reveals O-GlcNAcylation of mitochondrial voltage-dependent anion channel 2.

Protein modification by O-linked β-N-acetylglucosamine (O-GlcNAc) is a critical cell signaling modality, but identifying signal-specific O-GlcNAcylation events remains a significant experimental challenge. Here, we describe a method for visualizing and analyzing organelle- and stimulus-specific O-GlcNAcylated proteins and use it to identify the mitochondrial voltage-dependent anion channel 2 (VDAC2) as an O-GlcNAc substrate. VDAC2(-/-) cells resist the mitochondrial dysfunction and apoptosis caused by global O-GlcNAc perturbation, demonstrating a functional connection between O-GlcNAc signaling and mitochondrial physiology through VDAC2.

Predicting the emergence of the codling moth, Cydia pomonella (Lepidoptera: Tortricidae), on a degree-day scale in North America.

Background: Codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), is a major pest of apple, pear and walnut production in North America. Management programs are based on preventing larval entry into the fruit or nut and are typically timed by heat-driven models that are synchronized to field populations by first capture of overwintering moths in pheromone traps.

Toxicity of crude oil and pyrene to the embryos of beach spawning capelin (Mallotus villosus).

Due to a northward shift in oil and gas activities, there is an increasing need to understand the potential anthropogenic impacts of oil-related compounds on sub-Arctic and Arctic organisms, particularly those in coastal habitats. Capelin (Mallotus villosus), a key fish species in the Barents Sea ecosystem, undertakes aggregated spawning at both intertidal and subtidal coastal localities in northern Norway. To investigate the sensitivity of capelin embryos to oil compounds, newly fertilized capelin eggs were collected from a spawning beach and exposed until hatch (32 days) to either the water soluble fraction of crude oil or the single PAH compound, pyrene.

Effects of tether length on the behavior of amphiphilic bent-core molecules at water surfaces.

Alignment layers for bulk liquid crystalline phases can be created with monolayers formed by Langmuir-Schaefer techniques. Monolayer stability is a function of the propensity of the component molecule to effectively pack at a water interface; this propensity is enhanced when the molecule has an appropriate balance of hydrophilicity and hydrophobicity and the desired liquid crystalline order, as well as other structural factors. Our experiments show that molecules based on a bent-core with one hydrophilic and one hydrophobic end can form stable monolayers that act as effective alignment layers.

Determination of cell elasticity through hybrid ray optics and continuum mechanics modeling of cell deformation in the optical stretcher.

The optical stretcher is a dual-beam trap capable of stretching individual cells. Previous studies have used either ray- or wave-optical models to compute the optical pressure on the surface of a spherical cell. We have extended the ray-optics model to account for focusing by the spherical interface and the effects of multiple internal reflections.