Elevation of astrocyte-derived extracellular vesicles over the first month post-stroke in humans.

We sought to identify alterations in the quantity of plasma brain-derived extracellular vesicles (EV) over the first month post-stroke to shed light on related injury and repair mechanisms. We assessed plasma levels of presumed neuron-derived EVs (NDEs), astrocyte-derived EVs (ADEs), and oligodendrocyte-derived EVs (ODEs) in 58 patients 5, 15, and 30 days post-ischemic stroke and 46 controls matched for cardiovascular risk factors using sandwich immunoassays. Subsets of brain-derived EVs were identified by co-expression of the general EV marker CD9 and markers for neurons (L1CAM, CD171), astrocytes (EAAT1), and oligodendrocytes (MOG) respectively.

Centering Equity and Developing the Maternal Health Workforce: Building the National Maternal Health Learning and Innovation Center.

Purpose: The purpose of this article is to describe the development of the Maternal Health Learning and Innovation Center (MHLIC), a national initiative designed to enhance workforce capacity of maternal health professionals in the United States.

Description: The mission of the MHLIC is to foster collaboration and learning among diverse stakeholders to accelerate evidence-informed approaches advancing equitable maternal health outcomes through engagement, innovation, and policy. Working to center equity in all efforts, the MHLIC builds workforce capacity through partnership, training, technical assistance, coaching, facilitation of peer learning, and a national resource repository.

Evaluation and Management of Dysphagia in Amyotrophic Lateral Sclerosis: A Survey of Speech-Language Pathologists' Clinical Practice.

Objectives: The aim of this study was to determine the evaluation and management of dysphagia in amyotrophic lateral sclerosis (ALS) patients by speech-language pathologists (SLPs).

Methods: A 15-question web-based survey sent to SLPs in general clinical practice.

Results: Forty-nine SLPs responded.

Use cases, best practice and reporting standards for metabolomics in regulatory toxicology.

Metabolomics is a widely used technology in academic research, yet its application to regulatory science has been limited. The most commonly cited barrier to its translation is lack of performance and reporting standards. The MEtabolomics standaRds Initiative in Toxicology (MERIT) project brings together international experts from multiple sectors to address this need.

Use of non-invasive ventilation to facilitate extubation in a patient with amyotrophic lateral sclerosis with hypercapnic respiratory failure.

We present the case of a 44-year-old man with amyotrophic lateral sclerosis (ALS) intubated for hypercapnic respiratory failure and aspiration pneumonia. The patient was successfully extubated, transitioned to non-invasive ventilation and lived at home comfortably for 17 months, with good functional status for the first year. This case highlights the potential of prolonged survival post extubation in patients with advanced ALS and respiratory failure.

Vitronectin-Based, Biomimetic Encapsulating Hydrogel Scaffolds Support Adipogenesis of Adipose Stem Cells.

Soft tissue defects are relatively common, yet currently used reconstructive treatments have varying success rates, and serious potential complications such as unpredictable volume loss and reabsorption. Human adipose-derived stem cells (ASCs), isolated from liposuction aspirate have great potential for use in soft tissue regeneration, especially when combined with a supportive scaffold. To design scaffolds that promote differentiation of these cells down an adipogenic lineage, we characterized changes in the surrounding extracellular environment during adipogenic differentiation.

Histological characterization of human breast implant capsules.

Background: This study investigated the relationships between histomorphological aspects of breast capsules, including capsule thickness, collagen fiber alignment, the presence of α-smooth muscle actin (α-SMA)-positive myofibroblasts, and clinical observations of capsular contracture.

Methods: Breast capsule samples were collected at the time of implant removal in patients undergoing breast implant replacement or revision surgery. Capsular contracture was scored preoperatively using the Baker scale.

Skin extracellular matrix stimulation following injection of a hyaluronic acid-based dermal filler in a rat model.

Background: Hyaluronic acid-based dermal fillers have gained rapid acceptance for treating facial wrinkles and deep tissue folds. Although their space-filling properties are well understood, this study evaluates the cellular and molecular changes in skin, as a secondary effect, following injection of a commercially available, 24-mg/ml, cross-linked hyaluronic acid-based filler (HYC-24L+) in a rodent model.

Methods: Sprague-Dawley rats, aged 2 to 4 months, were injected intradermally with 20 μl of HYC-24L+ using a linear threading technique and followed to 12 weeks after injection.

Specific active immunotherapy of cancer: potential and perspectives.

Extensive research over the past two decades in tumor immunology has shown that immune reactivity to tumor antigens can restrict tumor growth and/or metastasis, especially when tumor burden is low. These observations in experimental models have been translated into clinical studies involving both active and passive forms of immunotherapies. While immune responses to specific tumor antigens can be detected in patients with various types of cancers, responses to any single antigen seldom correlate directly with a clinical response to tumors; however, some clinical regressions of solid tumors have been reported with certain types of cancer vaccines.

Targeting of humanized antibody D93 to sites of angiogenesis and tumor growth by binding to multiple epitopes on denatured collagens.

A humanized, affinity-matured IgG1 antibody, called D93, and its parental murine IgM HUI77 have been shown to specifically bind denatured collagens and thereby inhibit angiogenesis and tumor growth in various animal models. In this study, we have identified epitopes for both HUI77 and D93 on human collagen type IV. Several tryptic D93-binding peptides were identified by Western blot analysis and protein sequencing.

A novel source of viable peripheral blood mononuclear cells from leukoreduction system chambers.

Background: Buffy coats are becoming less available as a source of research-grade peripheral blood mononuclear cells (PBMNCs). Therefore, alternative sources of these cells were investigated.

Study Design And Methods: PBMNCs isolated from the cells retained in leukoreduction system chambers (LRSCs) and those eluted from white blood cell filters were compared.

Novel anti-denatured collagen humanized antibody D93 inhibits angiogenesis and tumor growth: An extracellular matrix-based therapeutic approach.

Matrix metalloproteases (MMPs) secreted by both tumor and endothelial cells proteolytically degrade collagen during tumor growth and neo-vascularization. This exposes cryptic binding sites on collagen with functional relevance for angiogenesis. In this report, we characterized a novel humanized monoclonal IgG1 antibody, D93.

Synergy among phytochemicals within crucifers: does it translate into chemoprotection?

The association between cruciferous vegetables and cancer prevention has been linked to glucosinolate derivatives. These phytochemicals enhance endogenous detoxification, leading to inactivation of potential carcinogens before initiation occurs. Two derivatives, indole-3-carbinol (I3C) and 1-cyano-2-hydroxy-3-butene (crambene) were shown in rats to induce a synergistic enhancement of detoxification enzyme activity.

Isoform-specific interactions of human apolipoprotein E to an intermediate conformation of human Alzheimer amyloid-beta peptide.

Brain plaque deposits of amyloid-beta peptide (Abeta) is a pathological hallmark of Alzheimer's disease (AD) and apolipoprotein E (apoE) is thought to be involved in its deposition. One hypothesis for the role of apoE in the pathogenesis of AD is that apoE may be involved in deposition or clearance of Abeta by direct protein-to-protein interaction. Lipidated apoE4 bound preferentially to an intermediate aggregated form of Abeta and formed two- to three-fold more binding complexes than isoforms apoE2 or apoE3.

Characterization of polyvalent allogeneic vaccines.

Allogeneic polyvalent vaccines have significant therapeutic and manufacturing advantages including: (i) the presence of multiple tumour-associated antigens; (ii) the potential benefit of viable but non-replicating cells which can provide persistent antigen presentation to the patient, and (iii) the ability to be consistently manufactured in large lots that can be used to treat multiple patients and be fully tested before release. Canvaxin is an example of a multi-cell allogeneic, polyvalent active immunotherapy and has been extensively tested in phase I and II clinical trials. Results of these clinical studies show a statistically significant increase in median and five-year survival of stage III and stage IV surgically resected patients with melanoma as compared to matched historical controls.

A slowly formed transient conformer of Abeta(1-40) is toxic to inward channels of dissociated hippocampal and cortical neurons of rats.

The mechanism by which amyloid peptide (Abeta(1-40)) produces effects on neurotransmission is currently unresolved. In initial experiments, using the patch-clamp technique, we found that 11.5 microM of preaggregated Abeta(1-40) altered the hippocampal neuron resting membrane potential and inhibited action potential firing.

Spontaneous aggregation and cytotoxicity of the beta-amyloid Abeta1-40: a kinetic model.

The time dependency of the spontaneous aggregation of the fibrillogenic beta-amyloid peptide, Abeta1-40, was measured by turbidity, circular dichroism, HPLC, and fluorescence polarization. The results by all methods were comparable and they were most consistent with a kinetic model where the peptide first slowly forms an activated monomeric derivative (AM), which is the only species able to initiate, by tetramerization, the formation of linear aggregates. The anti-Abeta antibody 6E10, raised against residues 1-17, at concentrations of 200-300 nM delayed significantly the aggregation of 50 microM amyloid peptide.

Physical methods to determine the binding mode of putative ligands for hepatitis C virus NS3 helicase.

Several small molecules identified by high-throughput screening (HTS) were evaluated for their ability to bind to a nonstructural protein 3 (NS3) helicase from hepatitis C virus (HCV). Equilibrium dissociation constants (K(d)'s) of the compounds for this helicase were determined using several techniques including an assay measuring the kinetics of isothermal enzyme denaturation at several concentrations of the test molecule. Effects of two nonhydrolyzable ATP analogs on helicase denaturation were measured as controls using the isothermal denaturation (ITD) assay.

The essential role of a free sulfhydryl group in blocking the cholesteryl site of cholesteryl ester transfer protein (CETP).

Cholesteryl ester transfer protein (CETP) has at least one unpaired sulfhydryl residue, which we have shown previously to be in or near the active site region. We investigated the location of this unpaired cysteine residue(s) of CETP using chemical modification with fluorescent sulfhydryl-specific reagents, limited proteolysis, and amino acid/sequence analysis. The kinetics of labeling CETP by either 2-(4'-maleimidylanilino)-naphthalene-6-sulfonic acid (MIANS) or acrylodan were followed by observing the increase in fluorescence of the bound probes.

Determination of ligand-MurB interactions by isothermal denaturation: application as a secondary assay to complement high throughput screening.

We used a temperature-jump isothermal denaturation procedure with various methods of detection to evaluate the quality of putative inhibitors of MurB discovered by high-throughput screening. Three optical methods of detection-ultraviolet hyperchromicity of absorbance, fluorescence of bound dyes, and circular dichroism-as well as differential scanning calorimetry were used to dissect the effects of two chemical compounds and a natural substrate on the enzyme. The kinetics of the denaturation process and binding of the compounds detected by quenching of flavin fluorescence were used to quantitate the dose dependencies of the ligand effects.

A competitive fluorescence assay to measure the reactivity of compounds.

A sensitive competitive method was developed for assessing the reactivity of compounds toward glutathione and toward thiols in general. The method employs the reaction of the fluorogenic reagent fluorescein-5-maleimide (FM) with glutathione (GSH) to generate a large increase in fluorescence emission. When the reaction is measured in the presence of a compound that competes with FM toward GSH, the rate constant for fluorescent product formation increases while the total amount of product formed at the end of the reaction decreases.

Use of anti-CD3 x anti-HER2/neu bispecific antibody for redirecting cytotoxicity of activated T cells toward HER2/neu+ tumors.

Relapse after adjuvant chemotherapy or high-dose chemotherapy with stem cell transplant for high-risk breast cancer remains high and new strategies that provide additional antitumor effects are needed. This report describes methods to generate highly effective HER2/neu-specific cytotoxic T cells by arming activated T cells with anti-CD3 x anti-HER2/neu bispecific antibody (BsAb). OKT3 and 9184 (anti-HER2) monoclonal antibodies (mAb) were conjugated and used to arm T cells that were subsequently tested in binding, cytotoxicity, and cytokine secretion assays.

The ligand affinity of proteins measured by isothermal denaturation kinetics.

An isothermal denaturation kinetic method was developed for identifying potential ligands of proteins and measuring their affinity. The method is suitable for finding ligands specific toward proteins of unknown function and for large-scale drug screening. It consists of analyzing the kinetics of isothermal denaturation of the protein-with and without the presence of potential specific ligands-as measured by long-wavelength fluorescent dyes whose quantum yield increases when bound to hydrophobic regions exposed upon unfolding of the proteins.

Clinical impact of ex vivo differentiated myeloid precursors after high-dose chemotherapy and peripheral blood progenitor cell rescue.

The infusion of ex vivo differentiated myeloid precursors may be able to shorten the period of obligatory neutropenia after high-dose chemotherapy and peripheral blood progenitor cell rescue by providing cells capable of differentiating to mature neutrophils within days of infusion. To test this hypothesis, 21 female patients with metastatic breast cancer underwent progenitor cell mobilization with cyclophosphamide, etoposide and G-CSF. CD34+ cells from one to two leukapheresis products were isolated and placed in suspension culture with a serum-free growth medium supplemented with PIXY321.