Intact Mass Proteomics Using a Proteoform Atlas.

Top-down proteomics, the characterization of intact proteoforms by tandem mass spectrometry, is the principal method for proteoform characterization in complex samples. Top-down proteomics relies on precursor isolation and subsequent gas-phase fragmentation to make proteoform identifications. While this strategy can produce highly detailed molecular information, the reliance on time-intensive tandem MS limits the speed with which proteoforms can be identified.

Identification of Host Proteins Involved in Hepatitis B Virus Genome Packaging.

A critical part of the hepatitis B virus (HBV) life cycle is the packaging of the pregenomic RNA (pgRNA) into nucleocapsids. While this process is known to involve several viral elements, much less is known about the identities and roles of host proteins in this process. To better understand the role of host proteins, we isolated pgRNA and characterized its protein interactome in cells expressing either packaging-competent or packaging-incompetent HBV genomes.

Defining Distinct RNA-Protein Interactomes of SARS-CoV-2 Genomic and Subgenomic RNAs.

Host RNA binding proteins recognize viral RNA and play key roles in virus replication and antiviral mechanisms. SARS-CoV-2 generates a series of tiered subgenomic RNAs (sgRNAs), each encoding distinct viral protein(s) that regulate different aspects of viral replication. Here, for the first time, we demonstrate the successful isolation of SARS-CoV-2 genomic RNA and three distinct sgRNAs (N, S, and ORF8) from a single population of infected cells and characterize their protein interactomes.

Defining distinct RNA-protein interactomes of SARS-CoV-2 genomic and subgenomic RNAs.

Host RNA binding proteins recognize viral RNA and play key roles in virus replication and antiviral defense mechanisms. SARS-CoV-2 generates a series of tiered subgenomic RNAs (sgRNAs), each encoding distinct viral protein(s) that regulate different aspects of viral replication. Here, for the first time, we demonstrate the successful isolation of SARS-CoV-2 genomic RNA and three distinct sgRNAs (N, S, and ORF8) from a single population of infected cells and characterize their protein interactomes.

Elucidating the RNA-Protein Interactomes of Target RNAs in Tissue.

RNA-protein interactions are key to many aspects of cellular homeostasis and their identification is important to understanding cellular function. Multiple strategies have been developed for the RNA-centric characterization of RNA-protein complexes. However, these studies have all been done in immortalized cell lines that do not capture the complexity of heterogeneous tissue samples.

Evaluation of plasma tau and neurofilament light chain biomarkers in a 12-year clinical cohort of human prion diseases.

Prion diseases are fatal neurodegenerative conditions with highly accurate CSF and imaging diagnostic tests, but major unmet needs for blood biomarkers. Using ultrasensitive immuno-assays, we measured tau and neurofilament light chain (NfL) protein concentrations in 709 plasma samples taken from 377 individuals with prion disease during a 12 year prospective clinical study, alongside healthy and neurological control groups. This provides an unprecedented opportunity to evaluate their potential as biomarkers.

Organic dyes in illuminated manuscripts: a unique cultural and historic record.

In this study, we successfully addressed the challenges posed by the identification of dyes in medieval illuminations. Brazilwood pigment lakes and orcein purple colours were unequivocally identified in illuminated manuscripts dated by art historians to be from the thirteenth to the fifteenth centuries and in the Fernão Vaz Dourado Atlas (sixteenth century). All three works were on a parchment support.

J. M. Despréaux' lichens from the Canary Islands and West Africa: an account of a 19th century collection found in an English archive.

This is an historical and descriptive account of 28 herbarium specimens, 27 lichens and an alga, found in the archives of Charles Chalcraft, a descendant of the Bedford family, who were dye manufacturers in Leeds, England, in the 19th century. The lichens comprise 13 different morphotypes collected in the Canary Islands and West Africa by the French botanist J. M.

Topical negative pressure wound therapy: a review of its role and guidelines for its use in the management of acute wounds.

Over the past two decades, topical negative pressure (TNP) wound therapy has gained wide acceptance as a genuine strategy in the treatment algorithm for a wide variety of acute and chronic wounds. Although extensive experimental and clinical evidence exists to support its use and despite the recent emergence of randomised control trials, its role and indications have yet to be fully determined. This article provides a qualitative overview of the published literature appertaining to the use of TNP therapy in the management of acute wounds by an international panel of experts using standard methods of appraisal.

Negative pressure wound dressing of the radial forearm donor site.

Donor site complications of the radial forearm are a significant cause of post-operative morbidity. 15 patients had radial forearm free tissue donor sites treated with split skin grafts and a negative pressure dressing. All grafts showed 100% take at 5 days.

Clinical experience with the negative pressure wound dressing.

Between 1997 and 1999, more than 300 patients have been treated using negative pressure wound dressings. The technique has been used successfully to prepare various acute, chronic or infected wounds to accept a skin graft or flap, and to promote graft take at difficult donor sites. The advantages include rapid healing by secondary intention, reduced time to skin grafting, an increase in the rate of graft take and a reduction in donor site complications.

Primary sensory neuron survival following targeted administration of nerve growth factor to an injured nerve.

Nerve injuries induce neurochemical changes within primary sensory neurons, including expression of neuropeptides, and a loss of a substantial proportion of the neurons may possibly be caused by a lack of neurotrophic support. In the present study the role of nerve growth factor (NGF) in preventing these changes was investigated in monkeys by giving NGF peripherally through a fibronectin (Fn) conduit. A sensory nerve (superficial radial) was transected and a gap of 5 mm was bridged with either autologous sural nerve graft (SNG), Fn, or Fn impregnated with NGF (Fn-NGF).

Nerve growth factor enhances nerve regeneration through fibronectin grafts.

Soluble fibronectin and nerve growth factor (NGF) promote axonal regeneration when placed in silicone tubes. We investigated the ability of orientated fibronectin mats to bind and release bioactive NGF and the possibility of augmenting axonal regeneration following axotomy by using fibronectin conduits impregnated with NGF. The release of NGF was quantified using a fluorometric ELISA and bioactivity confirmed with a neuronal culture bioassay.

Targeted delivery of nerve growth factor via fibronectin conduits assists nerve regeneration in control and diabetic rats.

This study examined the influence of fibronectin conduits joining two halves of a sectioned sciatic nerve, with and without preimpregnation with nerve growth factor, on regeneration in rats with streptozotocin-induced diabetes mellitus. Regeneration, measured morphometrically in fibres containing immunoreactivity to calcitonin gene-related peptide (CGRP) and GAP-43, was significantly less (P < 0.0001 for all comparisons) in diabetic rats with fibronectin mat grafts without nerve growth factor compared to similarly treated controls.

Different muscle graft denaturing methods and their use for nerve repair.

In an attempt to enhance recovery through denatured muscle grafts we have used different methods to denature muscle, either using dry heat in a microwave oven or by warming in sterile distilled water. Axonal regeneration was studied at intervals from day 5 to day 60 after insertion of the muscle grafts and compared to results found in frozen-thawed muscle grafts and in autologous nerve grafts used as controls. Axonal regeneration, Schwann cell behaviour and the degree of inflammation were quantified using immunohistochemical techniques and computerised image analysis.

Orientated mats of fibronectin as a conduit material for use in peripheral nerve repair.

This study introduces a new nerve conduit material consisting of orientated strands of the cell adhesive fibronectin. Axonal regeneration, Schwann cell behaviour and the degree of inflammation were quantified using immunohistochemical techniques and computerized image analysis. The results when fibronectin was used to bridge a 1 cm defect in rat sciatic nerve were compared to those with autologous nerve grafts and freeze-thawed muscle grafts used as controls.

Increased axonal regeneration over long nerve gaps using autologous nerve-muscle sandwich grafts.

Poor recovery is seen after repair of long defects in peripheral nerves when denatured muscle grafts or regeneration chambers providing physical support alone are used. The presence of Schwann cells and neurotrophic factors is required for axons to migrate significant distances. In this study we have used immunohistochemical techniques and axon counts to quantify the regeneration seen when 5 cm defects in rabbit sciatic nerve were repaired with a composite graft consisting of 2-3 mm lengths of fresh autologous nerve sandwiched between 1 cm frozen-thawed muscle grafts.

Neck swellings which mimic branchial cysts in HIV-positive patients.

A retrospective review of over 700 HIV-positive patients referred to the ENT department at St Mary's Hospital was undertaken. This revealed nine patients whose initial presentation was due to an upper neck swelling. The clinical and cytological results are discussed as well as consideration of the similarity of these lesions to branchial cysts, and the implication of the involvement of lymphoid tissue in the aetiology of branchial cysts.