Cellular and Biophysical Applications of Genetic Code Expansion.

Despite their diverse functions, proteins are inherently constructed from a limited set of building blocks. These compositional constraints pose significant challenges to protein research and its practical applications. Strategically manipulating the cellular protein synthesis system to incorporate novel building blocks has emerged as a critical approach for overcoming these constraints in protein research and application. In the past two decades, the field of genetic code expansion (GCE) has achieved significant advancements, enabling the integration of numerous novel functionalities into proteins across a variety of organisms. This technological evolution has paved the way for the extensive application of genetic code expansion across multiple domains, including protein imaging, the introduction of probes for protein research, analysis of protein-protein interactions, spatiotemporal control of protein function, exploration of proteome changes induced by external stimuli, and the synthesis of proteins endowed with novel functions. In this comprehensive Review, we aim to provide an overview of cellular and biophysical applications that have employed GCE technology over the past two decades.

In Vivo Human Lacrimal Gland Imaging Using an Ultrasound Biomicroscopy.

Purpose: In the present study, we introduce human lacrimal gland imaging using an ultrasound biomicroscopy (UBM) with a soft cover and show their findings. Methods: The representative UBM findings of palpebral lobes in seven subjects (4 with non-Sjögren dry eye syndrome, 1 with Sjögren syndrome, and 2 healthy subjects) were described in this study. To prolapse the palpebral lobe, the examiner pulled the temporal part of the upper eyelid in the superotemporal direction and directed the subject to look in the inferonasal direction. We scanned the palpebral lobes longitudinally and transversely using UBM. We used an Aviso UBM (Quantel Medical, Clermont-Ferrand, France) with a 50 MHz linear probe and ClearScan. Results: In UBM of two healthy subjects, the echogenicity of the lacrimal gland was lower than that of the sclera and homogeneous. But, the parenchyma of a patient with Sjögren dry eye syndrome was quite inhomogeneous compared to the healthy subjects. In two patients with dry eye syndrome, we were able to observe some lobules in the parenchyma. We could find excretory ducts running parallel at the surface of the longitudinal section in some subjects. In the longitudinal UBM scan of a subject, we observed a tubular structure at a depth of 1500 µm that was considered a blood vessel. It ran from the superonasal to the inferotemporal direction. In a subject, we observed a large cyst beneath the conjunctiva. Conclusions: Lacrimal gland imaging using UBM has both advantages of OCT and sonography, and could be useful for evaluating dry eye syndrome.

Comparative analysis of image quality and interchangeability between standard and deep learning-reconstructed T2-weighted spine MRI.

RATIONALE AND OBJECTIVES: MRI reconstruction of undersampled data using a deep learning (DL) network has been recently performed as part of accelerated imaging. Herein, we compared DL-reconstructed T2-weighted image (T2-WI) to conventional T2-WI regarding image quality and degenerative lesion detection. MATERIALS AND METHODS: Sixty-two patients underwent C-spine (n = 27) or L-spine (n = 35) MRIs, including conventional and DL-reconstructed T2-WI. Image quality was assessed with non-uniformity measurement and 4-scale grading of structural visibility. Three readers (R1, R2, R3) independently assessed the presence and types of degenerative lesions. Student t-test was used to compare non-uniformity measurements. Interprotocol and interobserver agreement of structural visibility was analyzed with Wilcoxon signed-rank test and weighted-κ values, respectively. The diagnostic equivalence of degenerative lesion detection between two protocols was assessed with interchangeability test. RESULTS: The acquisition time of DL-reconstructed images was reduced to about 21-58% compared to conventional images. Non-uniformity measurement was insignificantly different between the two images (p-value = 0.17). All readers rated DL-reconstructed images as showing the same or superior structural visibility compared to conventional images. Significantly improved visibility was observed at disk margin of C-spine (R1, p < 0.001; R2, p = 0.04) and dorsal root ganglia (R1, p = 0.03; R3, p = 0.02) and facet joint (R1, p = 0.04; R2, p < 0.001; R3, p = 0.03) of L-spine. Interobserver agreements of image quality were variable in each structure. Clinical interchangeability between two protocols for degenerative lesion detection was verified showing <5% in the upper bounds of 95% confidence intervals of agreement rate differences. CONCLUSIONS: DL-reconstructed T2-WI demonstrates comparable image quality and diagnostic performance with conventional T2-WI in spine imaging, with reduced acquisition time.

New Korean Guidelines for the Diagnosis and Management of Dry Eye Disease.

New Korean guidelines for the diagnosis and management of dry eye disease were developed based on literature reviews by the Korean Dry Eye Guideline Establishment Committee, a previous dry eye guideline by Korean Corneal Disease Study Group, a survey of Korean Dry Eye Society (KDES) members, and KDES consensus meetings. The new definition of dry eye was also proposed by KDES regular members. The new definition by the regular members of the KDES is as follows: "Dry eye is a disease of the ocular surface characterized by tear film abnormalities and ocular symptoms." The combination of ocular symptoms and an unstable tear film (tear breakup time <7 seconds) was considered as essential components for the diagnosis of dry eye. Schirmer test and ocular surface staining were considered adjunctive diagnostic criteria. The treatment guidelines consisted of a simplified stepwise approach according to aqueous deficiency dominant, evaporation dominant, and altered tear distribution subtypes. New Korean guidelines can be used as a simple, valid, and accessible tool for the diagnosis and management of dry eye disease in clinical practice.

Engineering of cell-surface receptors for analysis of receptor internalization and detection of receptor-specific glycosylation.

The epidermal growth factor receptor (EGFR) is a cell-surface glycoprotein that is involved mainly in cell proliferation. Overexpression of this receptor is intimately related to the development of a broad spectrum of tumors. In addition, glycans linked to the EGFR are known to affect its EGF-induced activation. Because of the pathophysiological significance of the EGFR, we prepared a fluorescently labeled EGFR (EGFR128-AZDye 488) on the cell surface by employing the genetic code expansion technique and bioorthogonal chemistry. EGFR128-AZDye 488 was initially utilized to investigate time-dependent endocytosis of the EGFR in live cells. The results showed that an EGFR inhibitor and antibody suppress endocytosis of the EGFR promoted by the EGF, and that lectins recognizing glycans of the EGFR do not enhance EGFR internalization into cells. Observations made in studies of the effects of appended glycans on the entry of the EGFR into cells indicate that a de-sialylated or de-fucosylated EGFR is internalized into cells more efficiently than a wild-type EGFR. Furthermore, by using the FRET-based imaging method of cells which contain an EGFR linked to AZDye 488 (a FRET donor) and cellular glycans labeled with rhodamine (a FRET acceptor), sialic acid residues attached to the EGFR were specifically detected on the live cell surface. Taken together, the results suggest that a fluorescently labeled EGFR will be a valuable tool in studies aimed at gaining an understanding of cellular functions of the EGFR.

Affinity-Directed Site-Specific Protein Labeling and Its Application to Antibody-Drug Conjugates.

Chemically modified proteins have diverse applications; however, conventional chemo-selective methods often yield heterogeneously labeled products. To address this limitation, site-specific protein labeling holds significant potential, driving extensive research in this area. Nevertheless, site-specific modification of native proteins remains challenging owing to the complexity of their functional groups. Therefore, a method for site-selective labeling of intact proteins is aimed to design. In this study, a novel approach to traceless affinity-directed intact protein labeling is established, which leverages small binding proteins and genetic code expansion technology. By applying this method, a site-specific antibody labeling with a drug, which leads to the production of highly effective antibody-drug conjugates specifically targeting breast cancer cell lines is achieved. This approach enables traceless conjugation of intact target proteins, which is a critical advantage in pharmaceutical applications. Furthermore, small helical binding proteins can be easily engineered for various target proteins, thereby expanding their potential applications in diverse fields. This innovative approach represents a significant advancement in site-specific modification of native proteins, including antibodies. It also bears immense potential for facilitating the development of therapeutic agents for various diseases.

Deep learning-based k-space-to-image reconstruction and super resolution for diffusion-weighted imaging in whole-spine MRI.

PURPOSE: To assess the feasibility of deep learning (DL)-based k-space-to-image reconstruction and super resolution for whole-spine diffusion-weighted imaging (DWI). METHOD: This retrospective study included 97 consecutive patients with hematologic and/or oncologic diseases who underwent DL-processed whole-spine MRI from July 2022 to March 2023. For each patient, conventional (CONV) axial single-shot echo-planar DWI (b = 50, 800 s/mm2) was performed, followed by DL reconstruction and super resolution processing. The presence of malignant lesions and qualitative (overall image quality and diagnostic confidence) and quantitative (nonuniformity [NU], lesion contrast, signal-to-noise ratio [SNR], contrast-to-noise ratio [CNR], and ADC values) parameters were assessed for DL and CONV DWI. RESULTS: Ultimately, 67 patients (mean age, 63.0 years; 35 females) were analyzed. The proportions of vertebrae with malignant lesions for both protocols were not significantly different (P: [0.55-0.99]). The overall image quality and diagnostic confidence scores were higher for DL DWI (all P ≤ 0.002) than CONV DWI. The NU, lesion contrast, SNR, and CNR of each vertebral segment (P ≤ 0.04) but not the NU of the sacral segment (P = 0.51) showed significant differences between protocols. For DL DWI, the NU was lower, and lesion contrast, SNR, and CNR were higher than those of CONV DWI (median values of all segments; 19.8 vs. 22.2, 5.4 vs. 4.3, 7.3 vs. 5.5, and 0.8 vs. 0.7). Mean ADC values of the lesions did not significantly differ between the protocols (P: [0.16-0.89]). CONCLUSIONS: DL reconstruction can improve the image quality of whole-spine diffusion imaging.

Accelerating acquisition of readout-segmented echo planar imaging (rs-EPI) with a simultaneous multislice (SMS) technique for diffusion-weighted (DW) breast MRI: Evaluation of image quality factors.

PURPOSE: This study aims to compare the image quality, apparent diffusion coefficient (ADC), signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR) values, and scan time between readout-segmented echo planar imaging (rs-EPI) and simultaneous multislice (SMS) rs-EPI sequences. METHODS: A total of 80 consecutive women who underwent breast diffusion-weighted imaging (DWI) were included, and two rs-EPI DWI sequences with and without SMS were acquired and compared. Qualitative analysis involved three radiologists independently scoring image quality and radiologist preference. For quantitative comparison, the radiologists independently measured the ADC values in patients, while SNR, CNR, and ADC values were measured on a phantom. RESULTS: The acquisition time was 5:47 min for rs-EPI and 3:20 min for SMS rs-EPI. In terms of image quality, scores were similar between rs-EPI and SMS rs-EPI sequences in the pooled data set, with the exception of skin-line distinction (p = 0.001) and background noise (p < 0.001). All radiologists considered SMS rs-EPI as equal or superior to rs-EPI in more than 70 % of cases. SMS rs-EPI demonstrated significantly higher ADC values than rs-EPI by all radiologists (p ≤ 0.002). For the phantom measurement, ADC (SMS: 1.26 ± 0.68 and RS: 1.26 ± 0.68, p = 0.198), SNR (SMS: 540.6 ± 342.1 and RS: 558.8 ± 523.2, p = 0.927), and CNR (SMS: 235.5 ± 38.9 and RS: 252.8 ± 108.0, p = 0.784) values did not significantly differ between the two sequences. CONCLUSION: SMS rs-EPI exhibited comparable image quality and similar ADC, SNR, and CNR values to rs-EPI while reducing the scan time by 42%.

A closer look into the cellular and molecular biology of myoepithelial cells across various exocrine glands.

Myoepithelial cells (MECs) are a unique subset of epithelial cells that possess several smooth muscle cell characteristics, such as a high number of actin-myosin filaments and the ability to contract. These cells are primarily located around the secretory cells of exocrine glands, including the salivary, mammary, lacrimal, and sweat glands. Their primary functions involve the construction of the basement membrane and help with secretion of gland products through contraction. So far, no comparative analysis of MECs in different exocrine glands had ever evaluated their differences. In this review, we took advantage of the various publicly available scRNAseq data from mouse exocrine glands to identify their shared and unique characteristics. The aim of this review is to compare the role of MECs in maintaining healthy glandular function, their involvement in disease states, and their regenerative capacity, with a particular emphasis on the latest research findings in these areas.

Distortion correction using topup algorithm by single k-space (TASK) for echo planar imaging.

Distortion of echo planar imaging (EPI) can be corrected using B0 field maps, which can be estimated with the topup algorithm that requires two EPI images with opposite distortions. In this study, we propose a new algorithm, termed topup algorithm by single K-space (TASK), to generate two input images from a single k-space for the topup algorithm to correct EPI distortions. The centric EPI contains the opposite phase-encoding polarities in one k-space, which can be divided into two halves with opposite distortions. Therefore, two inputs could be extracted by dividing the k-space into halves and processing them using the proposed procedure including an iterative procedure of automatic brain masking and uniformity correction. The efficiency of TASK was evaluated using 3D EPI. Quantitative evaluations showed that TASK corrected EPI distortion at a similar level to the traditional methods. The estimated field maps from the conventional topup and TASK showed a high correlation ([Formula: see text]). An ablation study showed the validity of every suggested step. Furthermore, it was confirmed that TASK was effective for distortion correction of two-shot centric EPI as well, demonstrating its wider applicability. In conclusion, TASK can correct EPI distortions by its own single k-space information with no additional scan.

Local Myeloid-Derived Suppressor Cells Impair Progression of Experimental Autoimmune Uveitis by Alleviating Oxidative Stress and Inflammation.

Purpose: To evaluate the immune regulatory effect of human cord blood myeloid-derived suppressor cells (MDSCs) in experimental autoimmune uveitis (EAU) models. Methods: MDSCs (1 × 106) or PBS were injected into established C57BL/6 EAU mice via the subconjunctival route on days 0 and 7. The severity of intraocular inflammation was evaluated for up to 3 weeks. Tissue injury and inflammation were analyzed using immunolabelled staining, real-time PCR, and ELISA. In addition, immune cells in draining lymph nodes (LNs) were quantified using flow cytometry. Results: After 21 days, the clinical scores and histopathological grades of EAU were lower in the MDSCs group compared with the PBS group. Local administration of MDSCs suppressed the oxidative stress and the expression of TNF-α and IL-1ß in the retinal tissues. In addition, it inhibited the activation of pathogenic T helper 1 (Th1) and Th17 cells in draining LNs. MDSCs increased the frequency of CD25+ Foxp3+ regulatory T cells and the mRNA expression of IL-10, as an immune modulator. Conclusions: MDSCs suppressed inflammation and oxidative stress in the retina and inhibited pathogenic T cells in the LNs in EAU. Therefore, ocular administration of MDSCs has therapeutic potential for uveitis.

The First Transcriptomic Atlas of the Adult Lacrimal Gland Reveals Epithelial Complexity and Identifies Novel Progenitor Cells in Mice.

The lacrimal gland (LG) secretes aqueous tears. Previous studies have provided insights into the cell lineage relationships during tissue morphogenesis. However, little is known about the cell types composing the adult LG and their progenitors. Using scRNAseq, we established the first comprehensive cell atlas of the adult mouse LG to investigate the cell hierarchy, its secretory repertoire, and the sex differences. Our analysis uncovered the complexity of the stromal landscape. Epithelium subclustering revealed myoepithelial cells, acinar subsets, and two novel acinar subpopulations: Tfrchi and Car6hi cells. The ductal compartment contained Wfdc2+ multilayered ducts and an Ltf+ cluster formed by luminal and intercalated duct cells. Kit+ progenitors were identified as: Krt14+ basal ductal cells, Aldh1a1+ cells of Ltf+ ducts, and Sox10+ cells of the Car6hi acinar and Ltf+ epithelial clusters. Lineage tracing experiments revealed that the Sox10+ adult populations contribute to the myoepithelial, acinar, and ductal lineages. Using scRNAseq data, we found that the postnatally developing LG epithelium harbored key features of putative adult progenitors. Finally, we showed that acinar cells produce most of the sex-biased lipocalins and secretoglobins detected in mouse tears. Our study provides a wealth of new data on LG maintenance and identifies the cellular origin of sex-biased tear components.

The Degree of Hydronephrosis as an Indicator of the Necessity for Ureteric Dilatation during Ureteroscopic Lithotripsy.

During rigid ureteroscopic lithotripsy, it is often encountered that the ureter is difficult to access. Attempts to advance the ureteroscope make the surgery more difficult. This study evaluated the preoperative predictive factors associated with difficult ureteral access (difficult ureter (DU)) during URS and assessed if clinical outcomes differed according to the degree of DU. This study identified 217 patients who underwent rigid ureteroscopic (URS) lithotripsy for the management of ureter stones between June 2017 and July 2021 in a tertiary hospital in Korea. In this group, preoperative factors were identified using univariate and multiple logistic regression analyses that could predict the degree of DU. Additionally, we also evaluated differences in treatment outcomes depending on the degree of DU. In 50 URS cases (22.0%), ureteral access using a ureteroscope was difficult. In the univariate and multivariate analyses, the degree of hydronephrosis was associated with the degree of DU. Treatment outcomes, extended operation times, low stone-free rate, postoperative pain, and secondary treatment were also significantly associated with the degree of DU. Clinicians can counsel patients with a lesser degree of hydronephrosis and approach their management accordingly.

Single-cell transcriptomics suggest distinct upstream drivers of IL-17A/F in hidradenitis versus psoriasis.

BACKGROUND: On the basis of the mounting evidence that type 17 T (T17) cells and increased IL-17 play a key role in driving hidradenitis suppurativa (HS) lesion development, biologic agents used previously in psoriasis that block signaling of IL-17A and/or IL-17F isoforms have been repurposed to treat HS. OBJECTIVE: Our research aimed to characterize the transcriptome of HS T17 cells compared to the transcriptome of psoriasis T17 cells, along with their ligand-receptor interactions with neighborhood immune cell subsets. METHODS: Single-cell data of 12,300 cutaneous immune cells from 8 deroofing surgical HS skin samples including dermal tunnels were compared to single-cell data of psoriasis skin (19,525 cells from 11 samples) and control skin (11,920 cells from 10 samples). All single-cell data were generated by the same protocol. RESULTS: HS T17 cells expressed lower levels of IL23R and higher levels of IL1R1 and IL17F compared to psoriasis T17 cells (P < .05). HS Treg cells expressed higher levels of IL1R1 and IL17F compared to psoriasis Treg cells (P < .05). Semimature dendritic cells were the major immune cell subsets expressing IL1B in HS, and IL-1ß ligand-receptor interactions between semimature dendritic cells and T17 cells were increased in HS compared to psoriasis (P < .05). HS dermal tunnel keratinocytes expressed inflammatory cytokines (IL17C, IL1A, IL1B, and IL6) that differed from the HS epidermis keratinocytes (IL36G) (P < .05). IL6, which synergizes with IL1B to maintain cytokine expression in T17 cells, was mainly expressed by fibroblasts in HS, which also expressed IL11+ inflammatory fibroblast genes (IL11, IL24, IL6, and POSTN) involved in the paracrine IL-1/IL-6 loop. CONCLUSION: The IL-1ß-T17 cell cytokine axis is likely a dominant pathway in HS with HS T17 cells activated by IL-1ß signaling, unlike psoriasis T17 cells, which are activated by IL-23 signaling.

Synthesis and biological evaluation of atropisomeric tetrahydroisoquinolines overcoming docetaxel resistance in triple-negative human breast cancer cells.

Herein, atropisomeric 8-aryltetrahydroisoquinolines have been synthesized and biologically evaluated. Based on our structure-activity relationship study, a highly bioactive racemic compound has been produced, and it exhibited high antiproliferative activities against various cancer cell lines, including docetaxel-resistant breast cancer cell lines. Each enantiomer can be synthesized in an enantioselective manner by employing the chiral phosphoric acid-catalyzed atroposelective Pictet-Spengler cyclization. An axially (R)-configured enantiomer showed a higher biological activity compared with the axially (S)-configured enantiomer. Further biological studies suggested that the (R)-enantiomer overcomes docetaxel resistance via the downregulation of signal transducer and activator of transcription 3 activation and consequently induces cellular apoptosis in docetaxel-resistant triple-negative breast cancer cell lines.

Deep Learning-Accelerated Liver Diffusion-Weighted Imaging: Intraindividual Comparison and Additional Phantom Study of Free-Breathing and Respiratory-Triggering Acquisitions.

OBJECTIVES: Deep learning-reconstructed diffusion-weighted imaging (DL-DWI) is an emerging promising time-efficient method for liver evaluation, but analyses regarding different motion compensation strategies are lacking. This study evaluated the qualitative and quantitative features, sensitivity for focal lesion detection, and scan time of free-breathing (FB) DL-DWI and respiratory-triggered (RT) DL-DWI compared with RT conventional DWI (C-DWI) in the liver and a phantom. MATERIALS AND METHODS: Eighty-six patients indicated for liver MRI underwent RT C-DWI, FB DL-DWI, and RT DL-DWI with matching imaging parameters other than the parallel imaging factor and number of averages. Two abdominal radiologists independently assessed qualitative features (structural sharpness, image noise, artifacts, and overall image quality) using a 5-point scale. The signal-to-noise ratio (SNR) along with the apparent diffusion coefficient (ADC) value and its standard deviation (SD) were measured in the liver parenchyma and a dedicated diffusion phantom. For focal lesions, per-lesion sensitivity, conspicuity score, SNR, and ADC value were evaluated. Wilcoxon signed rank test and repeated-measures analysis of variance with post hoc test revealed the difference in DWI sequences. RESULTS: Compared with RT C-DWI, the scan times for FB DL-DWI and RT DL-DWI were reduced by 61.5% and 23.9%, respectively, with statistically significant differences between all 3 pairs (all P 's < 0.001). Respiratory-triggered DL-DWI showed a significantly sharper liver margin, less image noise, and more minor cardiac motion artifact compared with RT C-DWI (all P 's < 0.001), whereas FB DL-DWI showed more blurred liver margins and poorer intrahepatic vessels demarcation than RT C-DWI. Both FB- and RT DL-DWI showed significantly higher SNRs than RT C-DWI in all liver segments (all P 's < 0.001). There was no significant difference in overall ADC values across DWI sequences in the patient or phantom, with the highest value recorded in the left liver dome by RT C-DWI. The overall SD was significantly lower with FB DL-DWI and RT DL-DWI than RT C-DWI (all P 's ≤ 0.003). Respiratory-triggered DL-DWI showed a similar per-lesion sensitivity (0.96; 95% confidence interval, 0.90-0.99) and conspicuity score to those of RT C-DWI and significantly higher SNR and contrast-to-noise ratio values ( P ≤ 0.006). The per-lesion sensitivity of FB DL-DWI (0.91; 95% confidence interval, 0.85-0.95) was significantly lower than that of RT C-DWI ( P = 0.001), with a significantly lower conspicuity score. CONCLUSIONS: Compared with RT C-DWI, RT DL-DWI demonstrated superior SNR, comparable sensitivity for focal hepatic lesions, and reduced acquisition time, making it a suitable alternative to RT C-DWI. Despite FB DL-DWI's weakness in motion-related challenges, further refinement could potentiate FB DL-DWI in the context of abbreviated screening protocols, where time efficiency is a high priority.

Combining Transurethral Resection of Fibrous Tissue and Temporary Urethral Stent Insertion Is an Optimal Strategy for Minimally Invasive Treatment of Recurrent and Long Urethral Strictures.

This study investigated the optimal strategy for the treatment of chronic recurrent urethral strictures longer than 3 cm, using a temporary urethral stent. Between September 2011 and June 2021, 36 patients with chronic bulbomembranous urethral strictures underwent temporary urethral stent placement. Retrievable self-expandable polymer-coated bulbar urethral stents (BUSs) were placed in 21 patients (group A), and thermo-expandable nickel-titanium alloy urethral stents were placed in 15 patients (group M). Each group was subdivided into those with and without transurethral resection (TUR) of fibrotic scar tissue. The urethral patency rates at 1 year after stent removal were compared between the groups. The patients in group A showed a higher urethral patency maintenance rate at 1 year after stent removal than those in group M (81.0% vs. 40.0%, log rank test p = 0.012). Analysis of subgroups in which TUR was performed due to severe fibrotic scar, showed that the patients in group A showed a significantly higher patency rate than patients in group M (90.9% vs. 44.4%, log rank test p = 0.028). In the treatment of chronic urethral strictures with a long fibrotic scar, temporary BUS combined with TUR of fibrotic tissue seems to be the optimal minimally invasive treatment strategy.

An agonistic anti-Tie2 antibody suppresses the normal-to-tumor vascular transition in the glioblastoma invasion zone.

Tumor progression is intimately associated with the vasculature, as tumor proliferation induces angiogenesis and tumor cells metastasize to distant organs via blood vessels. However, whether tumor invasion is associated with blood vessels remains unknown. As glioblastoma (GBM) is featured by aggressive invasion and vascular abnormalities, we characterized the onset of vascular remodeling in the diffuse tumor infiltrating zone by establishing new spontaneous GBM models with robust invasion capacity. Normal brain vessels underwent a gradual transition to severely impaired tumor vessels at the GBM periphery over several days. Increasing vasodilation from the tumor periphery to the tumor core was also found in human GBM. The levels of vascular endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR2) showed a spatial correlation with the extent of vascular abnormalities spanning the tumor-invading zone. Blockade of VEGFR2 suppressed vascular remodeling at the tumor periphery, confirming the role of VEGF-VEGFR2 signaling in the invasion-associated vascular transition. As angiopoietin-2 (ANGPT2) was expressed in only a portion of the central tumor vessels, we developed a ligand-independent tunica interna endothelial cell kinase 2 (Tie2)-activating antibody that can result in Tie2 phosphorylation in vivo. This agonistic anti-Tie2 antibody effectively normalized the vasculature in both the tumor periphery and tumor center, similar to the effects of VEGFR2 blockade. Mechanistically, this antibody-based Tie2 activation induced VE-PTP-mediated VEGFR2 dephosphorylation in vivo. Thus, our study reveals that the normal-to-tumor vascular transition is spatiotemporally associated with GBM invasion and may be controlled by Tie2 activation via a novel mechanism of action.

Spatially and velocity-selective magnetization preparation for noncontrast-enhanced peripheral MR angiography.

The purpose of the current study was to develop spatially and velocity-selective (SVS) magnetization preparation pulses for noncontrast-enhanced peripheral MR angiography (MRA) to provide comparisons with velocity-selective (VS) MRA with comparison to velocity-selective (VS). VS preparation pulses were designed by concatenating multiple excitation steps, each of which was a combination of a hard RF pulse, VS unipolar gradient pulses, and refocusing RF pulses. SVS preparation pulses were designed by replacing the hard RF pulse with a sinc-shaped RF pulse combined with a symmetric tripolar gradient pulse (which does not perturb the velocity encoding by the VS unipolar gradient pulses). Numerical simulations were performed to verify the intended hybrid excitation selectivity of SVS pulses taking account of tissue relaxation, magnetic field errors, and eddy currents. In vivo experiments were performed in healthy subjects to verify the hybrid excitation selectivity, as well as to demonstrate the visualization of the entire peripheral arteries using six-station protocols. As demonstrated by numerical simulations, SVS preparation yielded a notch-shaped longitudinal magnetization (Mz )-velocity response within the spatial stopband (the same as VS preparation) and preserved the Mz of spins outside the stopband, regardless of its velocity. We confirmed these observations also through in vivo tests with good agreement in normalized arterial and muscle signal intensities. In six-station peripheral MRA experiments, the proposed SVS-MRA yielded significantly higher arterial signal-to-noise ratio (SNR) (51.6 ± 14.3 vs. 38.9 ± 10.9; p < 0.001) and contrast-to-noise ratio (CNR) (41.2 ± 13.0 vs. 31.3 ± 10.5; p < 0.001) compared with VS-MRA. The proposed SVS-MRA improves arterial SNR and CNR compared with VS-MRA by mitigating undesired presaturation of arterial blood upstream the imaging field of view.

Clinical Outcomes of Endoscope-Assisted Pulmonary Endarterectomy for Chronic Thromboembolic Pulmonary Hypertension.

PURPOSE: Pulmonary thromboembolism is a potentially life-threatening condition in patients with heart disease; however, limited studies discussing long-term outcomes exist. This study aimed to investigate the long-term outcomes of pulmonary endarterectomy (PEA) in patients with chronic thromboembolic pulmonary hypertension (CTEPH), focusing on the improvement of functional class and right ventricular (RV) pressure. MATERIALS AND METHODS: Clinical data of patients with CTEPH were obtained from Yonsei Hospital between May 2012 and December 2021, and reviewed retrospectively. Twenty-six patients underwent endoscope-guided PEA during the study period, and the mean follow-up duration was 24.8±23.4 months. RESULTS: After PEA, most patients (88.5%) were weaned from inotropes without extracorporeal membrane oxygenation support during the first few days. Two patients (7.6%) had cerebrovascular accidents without neurological deficits. On echocardiography, the RV systolic pressure and tricuspid regurgitation grades significantly improved (p<0.001). Furthermore, the mean left ventricle end-diastolic diameter was significant increased (p=0.003), and the left ventricular end-systolic diameter increased (p<0.001). The median intensive care unit stay was 3.0±9.4 days, and median hospital stay 16.0±26.5 days. The 5-year survival rate was 95.5%, and the 5-year freedom rate of cardiac death was 100%. There was a marked improvement in New York Heart Association (NYHA) status (p<0.001). Cox regression suggested that the main pulmonary artery (MPA) involvement is a significant predictor of non-improvement in functional class post-PEA. CONCLUSION: Mortality rates are low and patients experience a marked improvement in NYHA class and health status after PEA. Moreover, MPA involvement may affect functional outcomes.