Self-channeling of a multi-Joule 10†µm picosecond pulse train through long distances in air.

In the long-wave infrared (LWIR) range, where, due to wavelength scaling, the critical power of Kerr self-focusing Pcr in air increases to 300-400 GW, we demonstrate that without external focusing a train of picosecond CO2 laser pulses can propagate in the form of a single several-centimeter diameter channel over hundreds of meters. The train of 10 µm pulses, for which the total energy ≥20 J is distributed over several near-terawatt picosecond pulses with a maximum power ≤2Pcr, is generated naturally during short pulse amplification in a CO2 laser. It is observed that the high-power 10 µm beam forms a large diameter "hot gas" channel in the ambient air with a ≥ 50 ms lifetime. Simulations of the experiment show that such filamentation-free self-channeling regime has low propagation losses and can deliver multi-Joule/TW-power LWIR pulses over km-scale distances.

Ultrasound frequency sonication facilitates high-throughput and uniform dissociation of cellular aggregates and tissues.

A sample preparation step involving dissociation of tissues into their component cells is often required to conduct analysis of nucleic acids and other constituents from tissue samples. Frequently, the extracellular matrix and cell-cell adhesions are disrupted via treatment with a chemical dissociating reagent or various mechanical forces. In this work, a new, high-throughput, multiplexed method of dissociating tissues and cellular aggregates into single cells using ultrasound frequency bath sonication is explored and characterized. Different operating parameters are evaluated, and a treatment protocol with potential for uniform, high-throughput tissue dissociation is compared to the existing best chemical and orbital plate shaking protocol. Metrics such as percent dissociation, cellular recovery, average aggregate size, proportion of various aggregate sizes, membrane circularity, and cellular viability are subsequently assessed and found to be favorable. In optimized conditions, 53 ±â€¯8% of 1 mm biopsy cores are dissociated within 30 min using sonication alone, surpassing leading high-throughput orbital plate shaking techniques five-fold. Chemical digestion is also 2 times more effective when complexed with sonication rather than orbital plate shaking. RNA content, quality, and expression are found to be superior to the standard protocol in terms of transcriptional preservation.

Using the Food and Drug Administration´s Sentinel System for surveillance of TB infection.

BACKGROUND: We examined patterns in care for individuals treated for latent TB infection (LTBI) in the US Food and Drug Administration´s Sentinel System.METHODS: Using administrative claims data, we identified patients who filled standard LTBI treatment prescriptions during 2008-2019. In these cohorts, we assessed LTBI testing, clinical management, and treatment duration.RESULTS: Among 113,338 patients who filled LTBI prescriptions, 80% (90,377) received isoniazid (INH) only, 19% (21,235) rifampin (RIF) only, and 2% (1,726) INH + rifapentine (RPT). By regimen, the proportion of patients with documented prior testing for TBI was 79%, 54%, and 91%, respectively. Median therapy duration was 84 days (IQR 35-84) for the 3-month once-weekly INH + RPT regimen, 60 days (IQR 30-100) for the 6- to 9-month INH regimen, and 30 days (IQR 2-60) for the 4-month RIF regimen.CONCLUSIONS: Among the cohorts, INH-only was the most commonly prescribed LTBI treatment. Most persons who filled a prescription for LTBI treatment did not have evidence of completing recommended treatment duration. These data further support preferential use of shorter-course regimens such as INH + RPT.

A Comparative Analysis of Deep Learning Models for Automated Cross-Preparation Diagnosis of Multi-Cell Liquid Pap Smear Images.

Routine Pap smears can facilitate early detection of cervical cancer and improve patient outcomes. The objective of this work is to develop an automated, clinically viable deep neural network for the multi-class Bethesda System diagnosis of multi-cell images in Liquid Pap smear samples. 8 deep learning models were trained on a publicly available multi-class SurePath preparation dataset. This included the 5 best-performing transfer learning models, an ensemble, a novel convolutional neural network (CNN), and a CNN + autoencoder (AE). Additionally, each model was tested on a novel ThinPrep Pap dataset to determine model generalizability across different liquid Pap preparation methods with and without Deep CORAL domain adaptation. All models achieved accuracies >90% when classifying SurePath images. The AE CNN model, 99.80% smaller than the average transfer model, maintained an accuracy of 96.54%. During consecutive training attempts, individual transfer models had high variability in performance, whereas the CNN, AE CNN, and ensemble did not. ThinPrep Pap classification accuracies were notably lower but increased with domain adaptation, with ResNet101 achieving the highest accuracy at 92.65%. This indicates a potential area for future improvement: development of a globally relevant model that can function across different slide preparation methods.

Electric-field facilitated rapid and efficient dissociation of tissues Into viable single cells.

Single-Cell Analysis is a growing field that endeavors to obtain genetic profiles of individual cells. Disruption of cell-cell junctions and digestion of extracellular matrix in tissues requires tissue-specific mechanical and chemical dissociation protocols. Here, a new approach for dissociating tissues into constituent cells is described. Placing a tissue biopsy core within a liquid-filled cavity and applying an electric field between two parallel plate electrodes facilitates rapid dissociation of complex tissues into single cells. Different solution compositions, electric field strengths, and oscillation frequencies are investigated experimentally and with COMSOL Multiphysics. The method is compared with standard chemical and mechanical approaches for tissue dissociation. Treatment of tissue samples at 100 V/cm 1 kHz facilitated dissociation of 95 ± 4% of biopsy tissue sections in as little as 5 min, threefold faster than conventional chemical-mechanical techniques. The approach affords good dissociation of tissues into single cells while preserving cell viability, morphology, and cell cycle progression, suggesting utility for sample preparation of tissue specimens for direct Single-Cell Analysis.

Caloric Restriction and Weight Loss Are Primary Factors in the Early Tissue-Specific Metabolic Changes After Bariatric Surgery.

OBJECTIVE: To evaluate changes in insulin sensitivity, hormone secretion, and hepatic steatosis immediately after caloric restriction, vertical sleeve gastrectomy (VSG), and Roux-en-Y gastric bypass (RYGB). RESEARCH DESIGN AND METHODS: Obese subjects were assessed for 1) insulin sensitivity with hyperinsulinemic-euglycemic clamp with glucose tracer infusion, 2) adipokine concentrations with serum and subcutaneous adipose interstitial fluid sampling, and 3) hepatic fat content with MRI before and 7-10 days after VSG, RYGB, or supervised caloric restriction. RESULTS: Each group exhibited an ∼5% total body weight loss, accompanied by similar improvements in hepatic glucose production and hepatic, skeletal muscle, and adipose tissue insulin sensitivity. Leptin concentrations in plasma and adipose interstitial fluid were equally decreased, and reductions in hepatic fat were similar. CONCLUSIONS: The improvements in insulin sensitivity and adipokine secretion observed early after bariatric surgery are replicated by equivalent caloric restriction and weight loss.

Bedside detection of intracranial midline shift using portable magnetic resonance imaging.

Neuroimaging is crucial for assessing mass effect in brain-injured patients. Transport to an imaging suite, however, is challenging for critically ill patients. We evaluated the use of a low magnetic field, portable MRI (pMRI) for assessing midline shift (MLS). In this observational study, 0.064 T pMRI exams were performed on stroke patients admitted to the neuroscience intensive care unit at Yale New Haven Hospital. Dichotomous (present or absent) and continuous MLS measurements were obtained on pMRI exams and locally available and accessible standard-of-care imaging exams (CT or MRI). We evaluated the agreement between pMRI and standard-of-care measurements. Additionally, we assessed the relationship between pMRI-based MLS and functional outcome (modified Rankin Scale). A total of 102 patients were included in the final study (48 ischemic stroke; 54 intracranial hemorrhage). There was significant concordance between pMRI and standard-of-care measurements (dichotomous, κ = 0.87; continuous, ICC = 0.94). Low-field pMRI identified MLS with a sensitivity of 0.93 and specificity of 0.96. Moreover, pMRI MLS assessments predicted poor clinical outcome at discharge (dichotomous: adjusted OR 7.98, 95% CI 2.07-40.04, p = 0.005; continuous: adjusted OR 1.59, 95% CI 1.11-2.49, p = 0.021). Low-field pMRI may serve as a valuable bedside tool for detecting mass effect.

Portable, bedside, low-field magnetic resonance imaging for evaluation of intracerebral hemorrhage.

Radiological examination of the brain is a critical determinant of stroke care pathways. Accessible neuroimaging is essential to detect the presence of intracerebral hemorrhage (ICH). Conventional magnetic resonance imaging (MRI) operates at high magnetic field strength (1.5-3 T), which requires an access-controlled environment, rendering MRI often inaccessible. We demonstrate the use of a low-field MRI (0.064 T) for ICH evaluation. Patients were imaged using conventional neuroimaging (non-contrast computerized tomography (CT) or 1.5/3 T MRI) and portable MRI (pMRI) at Yale New Haven Hospital from July 2018 to November 2020. Two board-certified neuroradiologists evaluated a total of 144 pMRI examinations (56 ICH, 48 acute ischemic stroke, 40 healthy controls) and one ICH imaging core lab researcher reviewed the cases of disagreement. Raters correctly detected ICH in 45 of 56 cases (80.4% sensitivity, 95%CI: [0.68-0.90]). Blood-negative cases were correctly identified in 85 of 88 cases (96.6% specificity, 95%CI: [0.90-0.99]). Manually segmented hematoma volumes and ABC/2 estimated volumes on pMRI correlate with conventional imaging volumes (ICC = 0.955, p = 1.69e-30 and ICC = 0.875, p = 1.66e-8, respectively). Hematoma volumes measured on pMRI correlate with NIH stroke scale (NIHSS) and clinical outcome (mRS) at discharge for manual and ABC/2 volumes. Low-field pMRI may be useful in bringing advanced MRI technology to resource-limited settings.

Optimization of a Clinically Relevant Chemical-Mechanical Tissue Dissociation Workflow for Single-Cell Analysis.

INTRODUCTION: While single-cell analysis technology has flourished, obtaining single cells from complex tissues continues to be a challenge. Current methods require multiple steps and several hours of processing. This study investigates chemical and mechanical methods for clinically relevant preparation of single-cell suspension from frozen biopsy cores of complex tissues. The developed protocol can be completed in 15 min. METHODS: Frozen bovine liver biopsy cores were normalized by weight, dimension, and calculated cellular composition. Various chemical reagents were tested for their capability to dissociate the tissue via confocal microscopy, hemocytometry and quantitative flow cytometry. Images were processed using ImageJ. Quantitative flow cytometry with gating analysis was also used for the analysis of dissociation. Physical modeling simulations were conducted in COMSOL Multiphysics. RESULTS: A rapid method for tissue dissociation was developed for single-cell analysis techniques. The results of this study show that a combination of 1% type-1 collagenase and pronase or hyaluronidase in 100 U/µL HBSS solution is the most effective at dissociating 2.5 mm thawed bovine liver biopsy cores in 15 min, with dissociation efficiency of 37-42% and viability >90% as verified using live MDA-MB-231 cancer cells. Cellular dissociation is significantly improved by adding a controlled mechanical force during the chemical process, to dissociate 93 ± 8% of the entire tissue into single cells. CONCLUSIONS: Understanding cellular dissociation in ex vivo tissues is essential to the development of clinically relevant dissociation workflows. Controlled mechanical force in combination with chemical treatment produces high quality tissue dissociation. This research is relevant to the understanding and assessment of tissue dissociation and the establishment of an automated preparatory workflow for single cell diagnostics. SUPPLEMENTARY INFORMATION: The online version of this article (10.1007/s12195-021-00667-y) contains supplementary material, which is available to authorized users.

Cold exposure induces dynamic, heterogeneous alterations in human brown adipose tissue lipid content.

Brown adipose tissue undergoes a dynamic, heterogeneous response to cold exposure that can include the simultaneous synthesis, uptake, and oxidation of fatty acids. The purpose of this work was to quantify these changes in brown adipose tissue lipid content (fat-signal fraction (FSF)) using fat-water magnetic resonance imaging during individualized cooling to 3 °C above a participant's shiver threshold. Eight healthy men completed familiarization, perception-based cooling, and MRI-cooling visits. FSF maps of the supraclavicular region were acquired in thermoneutrality and during cooling (59.5 ± 6.5 min). Brown adipose tissue regions of interest were defined, and voxels were grouped into FSF decades (0-10%, 10-20%…90-100%) according to their initial value. Brown adipose tissue contained a heterogeneous morphology of lipid content. Voxels with initial FSF values of 60-100% (P < 0.05) exhibited a significant decrease in FSF while a simultaneous increase in FSF occurred in voxels with initial FSF values of 0-30% (P < 0.05). These data suggest that in healthy young men, cold exposure elicits a dynamic and heterogeneous response in brown adipose tissue, with areas initially rich with lipid undergoing net lipid loss and areas of low initial lipid undergoing a net lipid accumulation.

MR fingerprinting with simultaneous T1, T2, and fat signal fraction estimation with integrated B0 correction reduces bias in water T1 and T2 estimates.

PURPOSE: MR fingerprinting (MRF) sequences permit efficient T1 and T2 estimation in cranial and extracranial regions, but these areas may include substantial fat signals that bias T1 and T2 estimates. MRI fat signal fraction estimation is also a topic of active research in itself, but may be complicated by B0 heterogeneity and blurring during spiral k-space acquisitions, which are commonly used for MRF. An MRF method is proposed that separates fat and water signals, estimates water T1 and T2, and accounts for B0 effects with spiral blurring correction, in a single sequence. THEORY AND METHODS: A k-space-based fat-water separation method is further extended to unbalanced steady-state free precession MRF with swept echo time. Repeated application of this k-space fat-water separation to demodulated forms of the measured data allows a B0 map and correction to be approximated. The method is compared with MRF without fat separation across a broad range of fat signal fractions (FSFs), water T1s and T2s, and under heterogeneous static fields in simulations, phantoms, and in vivo. RESULTS: The proposed method's FSF estimates had a concordance correlation coefficient of 0.990 with conventional measurements, and reduced biases in the T1 and T2 estimates due to fat signal relative to other MRF sequences by several hundred ms. The B0 correction improved the FSF, T1, and T2 estimation compared to those estimates without correction. CONCLUSION: The proposed method improves MRF water T1 and T2 estimation in the presence of fat and provides accurate FSF estimation with inline B0 correction.

Leptin Receptor Signaling in Sim1-Expressing Neurons Regulates Body Temperature and Adaptive Thermogenesis.

Leptin signals to regulate food intake and energy expenditure under conditions of normative energy homeostasis. The central expression and function of leptin receptor B (LepRb) have been extensively studied during the past two decades; however, the mechanisms by which LepRb signaling dysregulation contributes to the pathophysiology of obesity remains unclear. The paraventricular nucleus of the hypothalamus (PVN) plays a crucial role in regulating energy balance as well as the neuroendocrine axes. The role of LepRb expression in the PVN in regard to the regulation of physiological function of leptin has been controversial. The single-minded homolog 1 gene (Sim1) is densely expressed in the PVN and in parts of the amygdala, making Sim1-Cre mice a useful model for examining molecular mechanisms regulating PVN function. In this study, we characterized the physiological role of LepRb in Sim1-expressing neurons using LepRb-floxed × Sim1-Cre mice. Sim1-specific LepRb-deficient mice were surprisingly hypophagic on regular chow but gained more weight upon exposure to a high-fat diet than did their control littermates. We show that Sim1-specific deletion of a single LepRb gene copy caused decreased surface and core body temperatures as well as decreased energy expenditure in ambient room temperatures in both female and male mice. Furthermore, cold-induced adaptive (nonshivering) thermogenesis is disrupted in homozygous knockout mice. A defective thermoregulatory response was associated with defective cold-induced upregulation of uncoupling protein 1 in brown adipose tissue and reduced serum T4. Our study provides novel functional evidence supporting LepRb signaling in Sim1 neurons in the regulation of body weight, core body temperature, and cold-induced adaptive thermogenesis.

Trajectory optimized NUFFT: Faster non-Cartesian MRI reconstruction through prior knowledge and parallel architectures.

PURPOSE: The non-uniform fast Fourier transform (NUFFT) involves interpolation of non-uniformly sampled Fourier data onto a Cartesian grid, an interpolation that is slowed by complex, non-local data access patterns. A faster NUFFT would increase the clinical relevance of the plethora of advanced non-Cartesian acquisition methods. METHODS: Here we customize the NUFFT procedure for a radial trajectory and GPU architecture to eliminate the bottlenecks encountered when allowing for arbitrary trajectories and hardware. We call the result TRON, for TRajectory Optimized NUFFT. We benchmark the speed and accuracy TRON on a Shepp-Logan phantom and on whole-body continuous golden-angle radial MRI. RESULTS: TRON was 6-30× faster than the closest competitor, depending on test data set, and was the most accurate code tested. CONCLUSIONS: Specialization of the NUFFT algorithm for a particular trajectory yielded significant speed gains. TRON can be easily extended to other trajectories, such as spiral and PROPELLER. TRON can be downloaded at http://github.com/davidssmith/TRON.

Fat-Water Phantoms for Magnetic Resonance Imaging Validation: A Flexible and Scalable Protocol.

As new techniques are developed to image adipose tissue, methods to validate such protocols are becoming increasingly important. Phantoms, experimental replicas of a tissue or organ of interest, provide a low cost, flexible solution. However, without access to expensive and specialized equipment, constructing stable phantoms with high fat fractions (e.g., >50% fat fraction levels such as those seen in brown adipose tissue) can be difficult due to the hydrophobic nature of lipids. This work presents a detailed, low cost protocol for creating 5x 100 mL phantoms with fat fractions of 0%, 25%, 50%, 75%, and 100% using basic lab supplies (hotplate, beakers, etc.) and easily accessible components (distilled water, agar, water-soluble surfactant, sodium benzoate, gadolinium-diethylenetriaminepentacetate (DTPA) contrast agent, peanut oil, and oil-soluble surfactant). The protocol was designed to be flexible; it can be used to create phantoms with different fat fractions and a wide range of volumes. Phantoms created with this technique were evaluated in the feasibility study that compared the fat fraction values from fat-water magnetic resonance imaging to the target values in the constructed phantoms. This study yielded a concordance correlation coefficient of 0.998 (95% confidence interval: 0.972-1.00). In summary, these studies demonstrate the utility of fat phantoms for validating adipose tissue imaging techniques across a range of clinically relevant tissues and organs.

Phantom-based field maps for gradient nonlinearity correction in diffusion imaging.

Gradient coils in magnetic resonance imaging do not produce perfectly linear gradient fields. For diffusion imaging, the field nonlinearities cause the amplitude and direction of the applied diffusion gradients to vary over the field of view. This leads to site- and scan-specific systematic errors in estimated diffusion parameters such as diffusivity and anisotropy, reducing reliability especially in studies that take place over multiple sites. These errors can be substantially reduced if the actual scanner-specific gradient coil magnetic fields are known. The nonlinearity of the coil fields is measured by scanner manufacturers and used internally for geometric corrections, but obtaining and using the information for a specific scanner may be impractical for many sites that operate without special-purpose local engineering and research support. We have implemented an empirical field-mapping procedure using a large phantom combined with a solid harmonic approximation to the coil fields that is simple to perform and apply. Here we describe the accuracy and precision of the approach in reproducing manufacturer gold standard field maps and in reducing spatially varying errors in quantitative diffusion imaging for a specific scanner. Before correction, median B value error ranged from 33 - 41 relative to manufacturer specification at 100 mm from isocenter; correction reduced this to 0 - 4. On-axis spatial variation in the estimated mean diffusivity of an isotropic phantom was 2.2% - 4.1% within 60 mm of isocenter before correction, 0.5% - 1.6% after. Expected fractional anisotropy in the phantom was 0; highest estimated fractional anisotropy within 60 mm of isocenter was reduced from 0.024 to 0.012 in the phase encoding direction (48% reduction) and from 0.020 to 0.006 in the frequency encoding direction (72% reduction).

An Individualized, Perception-Based Protocol to Investigate Human Physiological Responses to Cooling.

Cold exposure, a known stimulant of the thermogenic effects of brown adipose tissue (BAT), is the most widely used method to study BAT physiology in adult humans. Recently, individualized cooling has been recommended to standardize the physiological cold stress applied across participants, but critical experimental details remain unclear. The purpose of this work was to develop a detailed methodology for an individualized, perception-based protocol to investigate human physiological responses to cooling. Participants were wrapped in two water-circulating blankets and fitted with skin temperature probes to estimate BAT activity and peripheral vasoconstriction. We created a thermoesthesia graphical user interface (tGUI) to continuously record the subject's perception of cooling and shivering status during the cooling protocol. The protocol began with a 15 min thermoneutral phase followed by a series of 10 min cooling phases and concluded when sustained shivering (>1 min duration) occurred. Researchers used perception of cooling feedback (tGUI ratings) to manually adjust and personalize the water temperature at each cooling phase. Blanket water temperatures were recorded continuously during the protocol. Twelve volunteers (ages: 26.2 ± 1.4 years; 25% female) completed a feasibility study to evaluate the proposed protocol. Water temperature, perception of cooling, and shivering varied considerably across participants in response to cooling. Mean clavicle skin temperature, a surrogate measure of BAT activity, decreased (-0.99°C, 95% CI: -1.7 to -0.25°C, P = 0.16) after the cooling protocol, but an increase in supraclavicular skin temperature was observed in 4 participants. A strong positive correlation was also found between thermoesthesia and peripheral vasoconstriction (ρ = 0.84, P < 0.001). The proposed individualized, perception-based protocol therefore has potential to investigate the physiological responses to cold stress applied across populations with varying age, sex, body composition, and cold sensitivity characteristics.

Long-wave infrared picosecond parametric amplifier based on Raman shifter technology.

A new method for a long-wave infrared (LWIR), picosecond difference frequency generation (DFG) source using one near-infrared laser and a Raman shifter is experimentally tested and characterized. The signal seed for DFG is a Stokes pulse generated via transient stimulated Raman scattering in a nonlinear medium with a Raman frequency in the 2-20 µm range. A study of the dynamics of the transient Raman regime in liquid C6D6 has shown that the efficiency of Stokes production can be increased and the central wavelength can be controlled by chirping the pump pulse in order to compensate for chirping caused by self-phase modulation. High energy, ≥3 µJ, picosecond pulses at 10.6 µm have been generated in a GaSe crystal pumped by 1 mJ pulses of 1060 nm light from a Nd:glass laser.

Untangling the R2* contrast in multiple sclerosis: A combined MRI-histology study at 7.0 Tesla.

T2*-weighted multi-echo gradient-echo magnetic resonance imaging and its reciprocal R2* are used in brain imaging due to their sensitivity to iron content. In patients with multiple sclerosis who display pathological alterations in iron and myelin contents, the use of R2* may offer a unique way to untangle mechanisms of disease. Coronal slices from 8 brains of deceased multiple sclerosis patients were imaged using a whole-body 7.0 Tesla MRI scanner. The scanning protocol included three-dimensional (3D) T2*-w multi-echo gradient-echo and 2D T2-w turbo spin echo (TSE) sequences. Histopathological analyses of myelin and iron content were done using Luxol fast blue and proteolipid myelin staining and 3,3'-diaminobenzidine tetrahydrochloride enhanced Turnbull blue staining. Quantification of R2*, myelin and iron intensity were obtained. Variations in R2* were found to be affected differently by myelin and iron content in different regions of multiple sclerosis brains. The data shall inform clinical investigators in addressing the role of T2*/R2* variations as a biomarker of tissue integrity in brains of MS patients, in vivo.

Ghost reduction in echo-planar imaging by joint reconstruction of images and line-to-line delays and phase errors.

PURPOSE: To correct line-to-line delays and phase errors in echo-planar imaging (EPI). THEORY AND METHODS: EPI-trajectory auto-corrected image reconstruction (EPI-TrACR) is an iterative maximum-likelihood technique that exploits data redundancy provided by multiple receive coils between nearby lines of k-space to determine and correct line-to-line trajectory delays and phase errors that cause ghosting artifacts. EPI-TrACR was efficiently implemented using a segmented FFT and was applied to in vivo brain data acquired at 7 T across acceleration (1×-4×) and multishot factors (1-4 shots), and in a time series. RESULTS: EPI-TrACR reduced ghosting across all acceleration factors and multishot factors, compared to conventional calibrated reconstructions and the PAGE method. It also achieved consistently lower ghosting in the time series. Averaged over all cases, EPI-TrACR reduced root-mean-square ghosted signal outside the brain by 27% compared to calibrated reconstruction, and by 40% compared to PAGE. CONCLUSION: EPI-TrACR automatically corrects line-to-line delays and phase errors in multishot, accelerated, and dynamic EPI. While the method benefits from additional calibration data for initialization, it was not a requirement for most reconstructions. Magn Reson Med 79:3114-3121, 2018. © 2017 International Society for Magnetic Resonance in Medicine.