Initial Experience of Using First-Person Wearable Video Recording Technology During Central Venous Catheter Placement in the Cardiac Operating Room.

OBJECTIVE: The aim of this study was to use wearable video-recording technology to measure precisely the timing of discrete events during perioperative central venous catheter (CVC) placements. DESIGN: A single-center, observational, exploratory study on the use of wearable video-recording technology during intraoperative CVC placement. SETTING: The study was conducted at a University Hospital. PARTICIPANTS: Clinical anesthesia residents, cardiothoracic anesthesia fellows, and attending anesthesiologists participated in this study. INTERVENTIONS: Participants were asked to use eye-tracking glasses prior to the placement of a CVC in the cardiac operating rooms. No other instruction was given to the participants. MEASUREMENTS AND MAIN RESULTS: The authors measured the total time to complete the CVC placement, phase-specific time, and specific times of interest. They compared these times across 3 training levels and tested differences with analysis of variance. The authors' findings indicated significant differences in total CVC placement time when the procedure included a pulmonary artery catheter insertion (1,170 ± 364, 923 ± 272, and 596 ± 226 seconds; F2,63 = 12.71, p < 0.0001). Additionally, they found differences in interval times and times of interest. The authors observed a reduction of variability with increasing experience during the CVC placement phase. CONCLUSIONS: In this observational study, the study authors describe their experience using first-person wearable video-recording technology to precisely measure the timing of discrete events during CVC placement by anesthesia residents and anesthesiologists. Future work will leverage the eye-tracking capabilities of the existing hardware to identify areas of inefficiency to develop actionable targets for interventions that could improve trainee performance and patient safety.

Mechanistic study of copper oxide, zinc oxide, cadmium oxide, and silver nanoparticles-mediated toxicity on the probiotic Lactobacillus reuteri.

The use of metal/metal oxide nanoparticles (NPs) in consumer products has increased dramatically. Accordingly, human exposure to these NPs has increased. Lactobacillus reuteri, a member of the beneficial gut microbiota, is essential for human health. In the present study, the toxic effect of three metal oxides (CuO, ZnO, and CdO) and one metal (Ag) NPs on L. reuteri were investigated in vitro. L. reuteri was susceptible to all the prepared NPs in a dose-dependent manner, visualized as an increase in the zones of inhibition and a significant reduction in the maximum specific growth rates (µmax). The minimal inhibitory concentrations were 5.8, 26, 560, and 560 µg/mL for CdO-, Ag-, ZnO-, and CuO-NPs, respectively, and the respective minimal bactericidal concentrations were 60, 70, 1500, and 1500 µg/mL. Electron microscopic examinations revealed the adsorption of the prepared NPs on L. reuteri cell surface, causing cell wall disruption and morphological changes. These changes were accompanied by significant leakage of cellular protein content by 214%, 191%, 112%, and 101% versus the untreated control when L. reuteri was treated with CdO-, Ag-, CuO-, and ZnO-NPs, respectively. NPs also induced oxidative damage, where the malondialdehyde level was significantly increased, and glutathione content was significantly decreased. Quantifying the DNA damage using comet assay showed that CuONPs had the maximum DNA tail length (8.2 px vs. 2.1 px for the control). While CdONPs showed the maximum percentage of DNA in tail (15.5% vs. 3.1%). This study provides a mechanistic evaluation of the NPs-mediated toxicity to a beneficial microorganism.

Characterizing chromatin packing scaling in whole nuclei using interferometric microscopy.

Chromatin is the macromolecular assembly containing the cell's genetic information, and its architectural conformation facilitates accessibility to activation sites and thus gene expression. We have developed an analytical framework to quantify chromatin structure with spectral microscopy. Chromatin structure can be described as a mass fractal, with packing scaling D up to specific genomic length scales. Considering various system geometries, we established a model to measure D with the interferometric technique partial wave spectroscopy (PWS) and validated the analysis using finite difference time domain to simulate the PWS system. Calculations of D were consistent with ground truth electron microscopy measurements, enabling a high-throughput, label-free approach to quantifying chromatin structure in the nanometer length scale regime.

In vivo broadband visible light optical coherence tomography probe enables inverse spectroscopic analysis.

We report the design and characterization of a 6 mm outer diameter pull-back circumferential scanning visible optical coherence tomography probe. The probe's large visible bandwidth (500-695 nm) allowed for inverse spectroscopic analysis and an axial resolution of ∼1.1 µm in tissue. We verify spectral imaging capabilities by measuring microsphere backscattering spectra and demonstrate in vivo spatial nanoscale characterization of tissue.

Origins of subdiffractional contrast in optical coherence tomography.

We demonstrate that OCT images quantify subdiffractional tissue structure. Optical coherence tomography (OCT) measures stratified tissue morphology with spatial resolution limited by the temporal coherence length. Spectroscopic OCT processing, on the other hand, has enabled nanoscale sensitive analysis, presenting an unexplored question: how does subdiffractional information get folded into the OCT image and how does one best analyze to allow for unambiguous quantification of ultrastructure? We first develop an FDTD simulation to model spectral domain OCT with nanometer resolution. Using this, we validate an analytical relationship between the sample statistics through the power spectral density (PSD) of refractive index fluctuations and three measurable quantities (image mean, image variance, and spectral slope), and have found that each probes different aspects of the PSD (amplitude, integral and slope, respectively). Finally, we found that only the spectral slope, quantifying mass scaling, is monotonic with the sample autocorrelation shape.

Nanoscale chromatin imaging and analysis platform bridges 4D chromatin organization with molecular function.

Extending across multiple length scales, dynamic chromatin structure is linked to transcription through the regulation of genome organization. However, no individual technique can fully elucidate this structure and its relation to molecular function at all length and time scales at both a single-cell level and a population level. Here, we present a multitechnique nanoscale chromatin imaging and analysis (nano-ChIA) platform that consolidates electron tomography of the primary chromatin fiber, optical super-resolution imaging of transcription processes, and label-free nano-sensing of chromatin packing and its dynamics in live cells. Using nano-ChIA, we observed that chromatin is localized into spatially separable packing domains, with an average diameter of around 200 nanometers, sub-megabase genomic size, and an internal fractal structure. The chromatin packing behavior of these domains exhibits a complex bidirectional relationship with active gene transcription. Furthermore, we found that properties of PDs are correlated among progenitor and progeny cells across cell division.

Light-driven formation and rupture of droplet bilayers.

We demonstrate the optical manipulation of nanoliter aqueous droplets containing surfactant or lipid molecules and immersed in an organic liquid using near-infrared light. The resulting emulsion droplets are manipulated using both the thermocapillary effect and convective fluid motion. Droplet-pair interactions induced in the emulsion upon optical initiation and control provide direct observations of the coalescence steps in intricate detail. Droplet-droplet adhesion (bilayer formation) is observed under several conditions. Selective bilayer rupture is also realized using the same infrared laser. The technique provides a novel approach to studying thin film drainage and interface stability in emulsion dynamics. The formation of stable lipid bilayers at the adhesion interface between interacting water droplets can provide an optical platform on which to build droplet-based lipid bilayer assays. The technique also has relevance to understanding and improving microfluidics applications by devising Petri dish-based droplet assays requiring no substrate fabrication.

Spectral contrast optical coherence tomography angiography enables single-scan vessel imaging.

Optical coherence tomography angiography relies on motion for contrast and requires at least two data acquisitions per pointwise scanning location. We present a method termed spectral contrast optical coherence tomography angiography using visible light that relies on the spectral signatures of blood for angiography from a single scan using endogenous contrast. We demonstrate the molecular sensitivity of this method, which enables lymphatic vessel, blood, and tissue discrimination.

Multimodal interference-based imaging of nanoscale structure and macromolecular motion uncovers UV induced cellular paroxysm.

Understanding the relationship between intracellular motion and macromolecular structure remains a challenge in biology. Macromolecular structures are assembled from numerous molecules, some of which cannot be labeled. Most techniques to study motion require potentially cytotoxic dyes or transfection, which can alter cellular behavior and are susceptible to photobleaching. Here we present a multimodal label-free imaging platform for measuring intracellular structure and macromolecular dynamics in living cells with a sensitivity to macromolecular structure as small as 20 nm and millisecond temporal resolution. We develop and validate a theory for temporal measurements of light interference. In vitro, we study how higher-order chromatin structure and dynamics change during cell differentiation and ultraviolet (UV) light irradiation. Finally, we discover cellular paroxysms, a near-instantaneous burst of macromolecular motion that occurs during UV induced cell death. With nanoscale sensitive, millisecond resolved capabilities, this platform could address critical questions about macromolecular behavior in live cells.

Single capillary oximetry and tissue ultrastructural sensing by dual-band dual-scan inverse spectroscopic optical coherence tomography.

Measuring capillary oxygenation and the surrounding ultrastructure can allow one to monitor a microvascular niche and better understand crucial biological mechanisms. However, capillary oximetry and pericapillary ultrastructure are challenging to measure in vivo. Here we demonstrate a novel optical imaging system, dual-band dual-scan inverse spectroscopic optical coherence tomography (D2-ISOCT), that, for the first time, can simultaneously obtain the following metrics in vivo using endogenous contrast: (1) capillary-level oxygen saturation and arteriolar-level blood flow rates, oxygen delivery rates, and oxygen metabolic rates; (2) spatial characteristics of tissue structures at length scales down to 30 nm; and (3) morphological images up to 2 mm in depth. To illustrate the capabilities of D2-ISOCT, we monitored alterations to capillaries and the surrounding pericapillary tissue (tissue between the capillaries) in the healing response of a mouse ear wound model. The obtained microvascular and ultrastructural metrics corroborated well with each other, showing the promise of D2-ISOCT for becoming a powerful new non-invasive imaging tool.

The effect of glenohumeral plane of elevation on supraspinatus subacromial proximity.

Shoulder pain is a common clinical problem affecting most individuals in their lifetime. Despite the high prevalence of rotator cuff pathology in these individuals, the pathogenesis of rotator cuff disease remains unclear. Position and motion related mechanisms of rotator cuff disease are often proposed, but poorly understood. The purpose of this study was to determine the impact of systematically altering glenohumeral plane on subacromial proximities across arm elevation as measures of tendon compression risk. Three-dimensional models of the humerus, scapula, coracoacromial ligament, and supraspinatus were reconstructed from MRIs in 20 subjects. Glenohumeral elevation was imposed on the humeral and supraspinatus tendon models for three glenohumeral planes, which were chosen to represent flexion, scapular plane abduction, and abduction based on average values from a previous study of asymptomatic individuals. Subacromial proximity was quantified as the minimum distance between the supraspinatus tendon and coracoacromial arch (acromion and coracoacromial ligament), the surface area of the supraspinatus tendon within 2 mm proximity to the coracoacromial arch, and the volume of intersection between the supraspinatus tendon and coracoacromial arch. The lowest modeled subacromial supraspinatus compression measures occurred during flexion at lower angles of elevation. This finding was consistent across all three measures of subacromial proximity. Knowledge of this range of reduced risk may be useful to inform future studies related to patient education and ergonomic design to prevent the development of shoulder pain and dysfunction.

Automated Cell Selection Using Support Vector Machine for Application to Spectral Nanocytology.

Partial wave spectroscopy (PWS) enables quantification of the statistical properties of cell structures at the nanoscale, which has been used to identify patients harboring premalignant tumors by interrogating easily accessible sites distant from location of the lesion. Due to its high sensitivity, cells that are well preserved need to be selected from the smear images for further analysis. To date, such cell selection has been done manually. This is time-consuming, is labor-intensive, is vulnerable to bias, and has considerable inter- and intraoperator variability. In this study, we developed a classification scheme to identify and remove the corrupted cells or debris that are of no diagnostic value from raw smear images. The slide of smear sample is digitized by acquiring and stitching low-magnification transmission. Objects are then extracted from these images through segmentation algorithms. A training-set is created by manually classifying objects as suitable or unsuitable. A feature-set is created by quantifying a large number of features for each object. The training-set and feature-set are used to train a selection algorithm using Support Vector Machine (SVM) classifiers. We show that the selection algorithm achieves an error rate of 93% with a sensitivity of 95%.

Decorin core protein (decoron) shape complements collagen fibril surface structure and mediates its binding.

Decorin is the archetypal small leucine rich repeat proteoglycan of the vertebrate extracellular matrix (ECM). With its glycosaminoglycuronan chain, it is responsible for stabilizing inter-fibrillar organization. Type I collagen is the predominant member of the fibrillar collagen family, fulfilling both organizational and structural roles in animal ECMs. In this study, interactions between decoron (the decorin core protein) and binding sites in the d and e(1) bands of the type I collagen fibril were investigated through molecular modeling of their respective X-ray diffraction structures. Previously, it was proposed that a model-based, highly curved concave decoron interacts with a single collagen molecule, which would form extensive van der Waals contacts and give rise to strong non-specific binding. However, the large well-ordered aggregate that is the collagen fibril places significant restraints on modes of ligand binding and necessitates multi-collagen molecular contacts. We present here a relatively high-resolution model of the decoron-fibril collagen complex. We find that the respective crystal structures complement each other well, although it is the monomeric form of decoron that shows the most appropriate shape complementarity with the fibril surface and favorable calculated energies of interaction. One molecule of decoron interacts with four to six collagen molecules, and the binding specificity relies on a large number of hydrogen bonds and electrostatic interactions, primarily with the collagen motifs KXGDRGE and AKGDRGE (d and e(1) bands). This work helps us to understand collagen-decorin interactions and the molecular architecture of the fibrillar ECM in health and disease.