High levels of triglyceride-rich lipoproteins (TGRLs) in blood are linked to development atherosclerosis, yet the mechanisms by which these particles initiate inflammation endothelium unknown. TGRL isolated from human plasma during postprandial state was examined for its capacity bind cultured aortic endothelial cells (HAECs) and alter acute inflammatory response tumor necrosis factor-alpha. HAECs were repetitively incubated with dietary freshly 2 hours per day 1 3 days mimic lipidemia....

We describe the development, validation, and application of a novel PDMS-based microfluidic device for imaging leukocyte interaction with biological substrate at defined shear force employing parallel plate geometry that optimizes experimental throughput while decreasing reagent consumption. The is vacuum bonded above standard 6-well tissue culture accommodates monolayer endothelial cells, thereby providing channel to directly observe kinetics adhesion under flow. Computational fluid...

We present an innovative centrifugal microfluidic immunoassay platform (SpinDx) to address the urgent biodefense and public health need for ultrasensitive point-of-care/incident detection of botulinum toxin. The simple, sample-to-answer approach is based on binding toxins antibody-laden capture particles followed by sedimentation through a density-media in disk quantification laser-induced fluorescence. A blind, head-to-head comparison study SpinDx versus gold-standard mouse bioassay...

Intracellular calcium flux is an early step in the signaling cascade that bridges ligation of selectin and chemokine receptors to activation adhesive motile functions during recruitment on inflamed endothelium. Calcium was imaged real time provided a means correlating events neutrophils rolling E-selectin stimulated by microfluidic chamber. Integrin dependent neutrophil arrest triggered tethering IL-8 seconds before rapid rise intracellular calcium, which followed onset pseudopod formation....

ABSTRACT Objective : Atherosclerosis is a focal disease that develops at sites of low and oscillatory shear stress in arteries. This study aimed to understand how endothelial cells sense gradient fluid transduce signals regulate membrane expression cell adhesion molecules monocyte recruitment. Methods Human aortic were stimulated with TNF‐α simultaneously exposed linear increased from 0 16 dyne/cm 2 . Cell molecule activation NFκ B quantified by immunofluorescence microscopy resolution the...

BACKGROUND Centrifugal “lab on a disk” microfluidics is promising avenue for developing portable, low-cost, automated immunoassays. However, the necessity of incorporating multiple wash steps results in complicated designs that increase time and sample/reagent volumes needed to run assays raises probability errors. We present proof principle disk-based microfluidic immunoassay technique processes blood samples without conventional steps. METHODS Microfluidic disks were fabricated from layers...

Abstract E-selectin is expressed by the vascular endothelium and binds flowing neutrophils in blood to facilitate their recruitment into underlying tissue at sites of inflammation. L-selectin on engaged undergoes rapid clustering then coalescence trailing edge polarizing cells. These processes are believed increase valency capacity signal CD18 integrin activity. Neutrophils, upon exiting microvasculature, down-regulate surface through ectodomain shedding a disintegrin metalloprotease 17...

Abstract Background The proliferation of decentralized sample collection models improves patient access and experience with laboratory medicine. However, many analytes have limited stability in liquid whole blood, serum, or plasma such that prompt transport under controlled temperature protection is crucial to analytical accuracy. Here we validate the Labcorp TrueTherm™ reusable shipper intended thermally protect small volume specimens. Methods Various TrueTherm were configured using...

Abstract Leukocyte capture on inflamed endothelium is facilitated by a shift in LFA-1 from low to high affinity that supports binding ICAM-1. bonds help anchor polymorphonuclear leukocytes (PMN) shear flow, and their redistribution the leading edge guides pseudopod formation, migration, extravasation. These events can be disrupted at plasma membrane stabilizing low- or intermediate-affinity state with allosteric small molecules. We hypothesized minimum dimeric bond formation between...

We present a novel "Lab-on-a-Disk" platform and demonstrate its capability for rapid sensitive measurement of vascular endothelial growth factor (VEGF) intended patients suffering from diabetic retinopathy (DR) age-related macular degeneration (AMD). This approach combines sedimentation principles applied to microspheres under centrifugal force with signal amplification using an enzyme fluorogenic substrate readout. The simple single channel per assay separates, washes concentrates...

P-Selectin glycoprotein ligand-1 (PSGL-1) is a mucin-like expressed on the surface of leukocytes that serves as major ligand for selectin family adhesion molecules and functions in leukocyte tethering rolling activated endothelium platelets. Previous studies have implicated highly conserved cytoplasmic domain PSGL-1 regulating outside-in signaling integrin activation. However, physically functionally interact with this are not completely defined. Using yeast two-hybrid screen bait, novel...

L-selectin (CD62L) amplifies neutrophil capture within the microvasculature at sites of inflammation. Activation by G protein-coupled stimuli or through ligation promotes clustering and serves to increase its adhesiveness, signaling, colocalization with beta(2)-integrins. Currently, little is known about molecular process regulating lateral mobility L-selectin. On stimulation, a progressive change takes place in organization plasma membrane, resulting membrane domains that are...

Semen quality assessment in population-based epidemiologic studies presents logistical and financial challenges due to reliance on centralised laboratory semen analysis. The Trak Male Fertility Testing System is an FDA-cleared validated at-home test for sperm concentration volume, with a research use only motility test. Here we evaluate the System's overall utility among men participating Pregnancy Study Online (PRESTO), web-based study of North American couples planning pregnancy.

Anaplasma phagocytophilum is an obligate intracellular bacterium that has evolved mechanisms to hijack polymorphonuclear neutrophil (PMN) receptors and signaling pathways bind, infect, multiply within the host cell. E-selectin upregulated during inflammation a requisite endothelial receptor supports PMN capture, rolling, activation of integrin-mediated arrest. Ligands expressed by mediate binding endothelium via include sialyl Lewis x (sLe )-expressing ligands such as P-selectin glycoprotein...

The increased demand for decentralized blood sample collection presents numerous operational challenges diagnostics providers. Sample degradation including hemolysis due to time, temperature, and handling between laboratory analysis leads limited test menus unreliable results. Here we introduce the lightweight, portable Labcorp TrueSpin™ rapid point-of-care separation using commercially available microvolume tubes. TrueSpin is a class I FDA-registered device designed untrained users....

Abstract Background While solutions enabling at-home self-collection of capillary blood samples are maturing, many analytes require separation and controlled shipping temperatures for stable analysis in centralized laboratories. Here we present results a pilot lay user study evaluating usability specimen quality collection with centrifugation refrigerated serum specimens. Methods N=45 consented subjects were mailed kits including supplies using Tasso+™ device, Labcorp TrueTherm™ reusable...

Abstract Background In the United States, an estimated 13% of women between ages 15 and 49 years, experience impaired reproductive potential (fecundity). Assessment fecundity typically requires in-person physician visit venous blood collection to evaluate hormone levels identify health conditions. Exploring less invasive, at-home sampling options could offer couples insights into their personal abilities, providing peace mind aiding in family planning. Methods Capillary blood, obtained using...