The distance of insertion points from wound margins in interrupted and vertical mattress sutures influences the tensile load capacity: An in vitro experimental study.

AIM: To determine the tensile load capacity (TLC) and the tearing characteristics for interrupted and vertical mattress sutures with different insertion points from the wound margin, and the effect of the bite size when using vertical mattress sutures. MATERIALS AND METHODS: A total of 120 gingiva and lining mucosa samples obtained from pig jaws were divided into groups according to the suturing technique (interrupted and vertical mattress sutures), distance of the insertion points from the wound margin (margin, 1, 3, and 5 mm) and bite size (1, 3, and 5 mm). The TLC of the suture and the tearing characteristics were evaluated using a tensile tester device. RESULTS: The TLC was significantly higher for vertical mattress sutures than for interrupted sutures regardless of the distance of the insertion points from the wound margin (intergroup p < .001). This distance significantly influenced the TLC for vertical mattress sutures (p < .05) but not for interrupted sutures (p > .05). Testing the tearing characteristics revealed that no tissue tearing occurred in groups when the insertion points were more than 3 mm from the wound margin. CONCLUSION: The TLC is higher for vertical mattress sutures than for interrupted sutures, and it increases when the insertion points are farther from the wound margin.

Optimization of a flow cytometry test for routine monitoring of B cell maturation antigen targeted CAR in peripheral blood.

Chimeric antigen receptor (CAR) modified T cell therapies targeting BCMA have displayed impressive activity in the treatment of multiple myeloma. There are currently two FDA licensed products, ciltacabtagene autoleucel and idecabtagene vicleucel, for treating relapsed and refractory disease. Although correlative analyses performed by product manufacturers have been reported in clinical trials, there are limited options for reliable BCMA CAR T detection assays for physicians and researchers looking to explore it as a biomarker for clinical outcome. Given the known association of CAR T cell expansion kinetics with toxicity and response, being able to quantify BCMA CAR T cells routinely and accurately in the blood of patients can serve as a valuable asset. Here, we optimized an accurate and sensitive flow cytometry test using a PE-conjugated soluble BCMA protein, with a lower limit of quantitation of 0.19% of CD3+ T cells, suitable for use as a routine assay for monitoring the frequency of BCMA CAR T cells in the blood of patients receiving either ciltacabtagene autoleucel or idecabtagene vicleucel.

Transcriptome Profiling of a Soybean Mutant with Salt Tolerance Induced by Gamma-ray Irradiation.

Salt stress is a significant abiotic stress that reduces crop yield and quality globally. In this study, we utilized RNA sequencing (RNA-Seq) to identify differentially expressed genes (DEGs) in response to salt stress induced by gamma-ray irradiation in a salt-tolerant soybean mutant. The total RNA library samples were obtained from the salt-sensitive soybean cultivar Kwangan and the salt-tolerant mutant KA-1285. Samples were taken at three time points (0, 24, and 72 h) from two tissues (leaves and roots) under 200 mM NaCl. A total of 967,719,358 clean reads were generated using the Illumina NovaSeq 6000 platform, and 94.48% of these reads were mapped to 56,044 gene models of the soybean reference genome (Glycine_max_Wm82.a2.v1). The DEGs with expression values were compared at each time point within each tissue between the two soybeans. As a result, 296 DEGs were identified in the leaves, while 170 DEGs were identified in the roots. In the case of the leaves, eight DEGs were related to the phenylpropanoid biosynthesis pathway; however, in the roots, Glyma.03G171700 within GmSalt3, a major QTL associated with salt tolerance in soybean plants, was differentially expressed. Overall, these differences may explain the mechanisms through which mutants exhibit enhanced tolerance to salt stress, and they may provide a basic understanding of salt tolerance in soybean plants.

Anti-TACI single and dual-targeting CAR T cells overcome BCMA antigen loss in multiple myeloma.

Chimeric Antigen Receptor (CAR) T cells directed to B cell maturation antigen (BCMA) mediate profound responses in patients with multiple myeloma, but most patients do not achieve long-term complete remissions. In addition, recent evidence suggests that high-affinity binding to BCMA can result in on-target, off-tumor activity in the basal ganglia and can lead to fatal Parkinsonian-like disease. Here we develop CAR T cells against multiple myeloma using a binder to targeting transmembrane activator and CAML interactor (TACI) in mono and dual-specific formats with anti-BCMA. These CARs have robust, antigen-specific activity in vitro and in vivo. We also show that TACI RNA expression is limited in the basal ganglia, which may circumvent some of the toxicities recently reported with BCMA CARs. Thus, single-targeting TACI CARs may have a safer toxicity profile, whereas dual-specific BCMA-TACI CAR T cells have potential to avoid the antigen escape that can occur with single-antigen targeting.

Immunogenetic Metabolomics Reveals Key Enzymes That Modulate CAR T-cell Metabolism and Function.

Immune evasion is a critical step of cancer progression that remains a major obstacle for current T cell-based immunotherapies. Hence, we investigated whether it is possible to genetically reprogram T cells to exploit a common tumor-intrinsic evasion mechanism whereby cancer cells suppress T-cell function by generating a metabolically unfavorable tumor microenvironment (TME). In an in silico screen, we identified ADA and PDK1 as metabolic regulators. We then showed that overexpression (OE) of these genes enhanced the cytolysis of CD19-specific chimeric antigen receptor (CAR) T cells against cognate leukemia cells, and conversely, ADA or PDK1 deficiency dampened this effect. ADA-OE in CAR T cells improved cancer cytolysis under high concentrations of adenosine, the ADA substrate, and an immunosuppressive metabolite in the TME. High-throughput transcriptomics and metabolomics analysis of these CAR T cells revealed alterations of global gene expression and metabolic signatures in both ADA- and PDK1-engineered CAR T cells. Functional and immunologic analyses demonstrated that ADA-OE increased proliferation and decreased exhaustion in CD19-specific and HER2-specific CAR T cells. ADA-OE improved tumor infiltration and clearance by HER2-specific CAR T cells in an in vivo colorectal cancer model. Collectively, these data unveil systematic knowledge of metabolic reprogramming directly in CAR T cells and reveal potential targets for improving CAR T-cell therapy.

Genetic Clarification of Auricularia heimuer Strains Bred and Cultivated in Korea Using the ITS and IGS1 rDNA Region Sequences.

Auricularia is one of the broadly cultivated edible mushrooms in Korea. Most of the Korean Auricularia strains used for cultivation and breeding are known as A. auricula-judae. Recently, this species has been reported to belong to a species complex. Therefore, this study was carried out to genetically clarify the bred and cultivated Korean A. auricula-judae strains. The internal transcribed spacer (ITS) and IGS1 rDNA region sequences were determined from 10 A. auricula-judae strains by PCR and sequencing. Variation in the nucleotide sequence and sequence length of the two rDNA regions were found among the seven A. auricula-judae strains. A maximum-likelihood (ML) phylogenetic tree based on the ITS sequences clearly placed all the 10 Korean A. auricula-judae strains in the A. heimuer clade of the A. auricula-judae complex. A. heimuer is diverged from A. auricula-judae. An ML phylogenetic tree based on the IGS1 sequences revealed the close relationship between Korean A. heimuer strains to Chinese A. heimuer strains. But each strain could be distinguishable by the IGS1 sequence. Furthermore, progeny strains in the seven Korean strains could be differentiated from their parental strains by the IGS1 sequence based phylogenetic tree. Our results are expected to be used to complement the distinction of domestic Auricularia cultivars.

Immunogenetic metabolomics revealed key enzymes that modulate CAR-T metabolism and function.

Immune evasion is a critical step of cancer progression that remains a major obstacle for current T cell-based immunotherapies. Hence, we seek to genetically reprogram T cells to exploit a common tumor-intrinsic evasion mechanism, whereby cancer cells suppress T cell function by generating a metabolically unfavorable tumor microenvironment (TME). Specifically, we use an in silico screen to identify ADA and PDK1 as metabolic regulators, in which gene overexpression (OE) enhances the cytolysis of CD19-specific CD8 CAR-T cells against cognate leukemia cells, and conversely, ADA or PDK1 deficiency dampens such effect. ADA -OE in CAR-T cells improves cancer cytolysis under high concentrations of adenosine, the ADA substrate and an immunosuppressive metabolite in the TME. High-throughput transcriptomics and metabolomics in these CAR-Ts reveal alterations of global gene expression and metabolic signatures in both ADA- and PDK1- engineered CAR-T cells. Functional and immunological analyses demonstrate that ADA -OE increases proliferation and decreases exhaustion in α-CD19 and α-HER2 CAR-T cells. ADA-OE improves tumor infiltration and clearance by α-HER2 CAR-T cells in an in vivo colorectal cancer model. Collectively, these data unveil systematic knowledge of metabolic reprogramming directly in CAR-T cells, and reveal potential targets for improving CAR-T based cell therapy. Synopsis: The authors identify the adenosine deaminase gene (ADA) as a regulatory gene that reprograms T cell metabolism. ADA-overexpression (OE) in α-CD19 and α-HER2 CAR-T cells increases proliferation, cytotoxicity, memory, and decreases exhaustion, and ADA-OE α-HER2 CAR-T cells have enhanced clearance of HT29 human colorectal cancer tumors in vivo .

Alveolar ridge regeneration in two-wall-damaged extraction sockets of an in vivo experimental model.

AIM: To determine the healing outcome following grafting with deproteinized porcine bone mineral (DPBM) with or without collagen membrane coverage in two-wall (both buccal and lingual)-damaged extraction sockets. MATERIALS AND METHODS: Distal roots of three mandibular premolars in six beagle dogs were extracted, and the whole buccal and lingual bony walls were surgically removed. Three treatment protocols were then applied according to the following group allocation: no graft (None), grafting DPBM (BG), and grafting DPBM with coverage by a collagen membrane (BG + M). Two observational periods (2 and 8 weeks) were used with the split-mouth design, and quantitative and qualitative analyses were performed by microcomputed tomography and histology. RESULTS: The dimensions of the alveolar ridge at both grafted sites (BG and BG + M) remained similar to those of the pristine ridge in the histologic and radiographic analyses, whereas the ungrafted sites (None) collapsed both vertically and horizontally. Both grafting protocols produced substantial bony regeneration, but the addition of a covering membrane enhanced the proportion of mineralized tissue within the augmented area, and the BG + M group also showed a significantly larger area of regenerated ridge than the None group (p < .05). CONCLUSIONS: Bone grafting with collagen membrane can maintain the alveolar ridge dimensions with substantial bone regeneration in a two-wall-damaged extraction socket.

Distance of insertion points in a mattress suture from the wound margin for ideal primary closure in alveolar mucosa: an in vitro experimental study.

PURPOSE: This study was conducted to determine how the distance of the near insertion points in a vertical mattress suture from the wound margin influences the pattern of primary closure in an in vitro experimental model. METHODS: Pairs of 180 porcine gingival and alveolar mucosa samples were harvested from 90 pig jaws and fixed to a specially designed model. A vertical mattress suture was performed with the near insertion point at 3 different distances from the wound margin (1-, 3-, and 5-mm) on both the gingival and mucosal samples (6 groups; n=30 for each group). The margin discrepancy and the presence of epithelium between the wound margins were measured on histologic slides. RESULTS: The margin discrepancy decreased significantly as the near insertion point became closer to the wound margin both in mucosal tissue (0.241±0.169 mm, 0.945±0.497 mm, and 1.306±0.773 mm for the 1-, 3-, and 5-mm groups, respectively) and in gingival tissue (0.373±0.304 mm, 0.698±0.431 mm, and 0.713±0.691 mm, respectively). The frequency of complications of wound margin adaptation reduced as the distance of the near insertion point from the wound margin decreased both in the mucosal and gingival tissues. CONCLUSIONS: Placing the near insertion point close to the wound margin enhances the precision of wound margin approximation/adaptation using a vertical mattress suture.

Glymphatic Dysfunction in Patients With End-Stage Renal Disease.

BACKGROUND: We aimed to compare glymphatic dysfunction between patients with end-stage renal disease (ESRD) and healthy controls and analyze the correlation between the glymphatic function and clinical characteristics using the diffusion tensor image analysis along with the perivascular space (DTI-ALPS) index. METHODS: We prospectively enrolled neurologically asymptomatic 49 patients with ESRD undergoing dialysis and 38 healthy controls. Diffusion tensor image was conducted using the same 3T scanner, and the DTI-ALPS index was calculated. We compared the DTI-ALPS index between the patients with ESRD and healthy controls. In addition, we conducted a correlation analysis between the clinical characteristics and DTI-ALPS index in patients with ESRD. RESULTS: There were significant differences in the DTI-ALPS index between patients with ESRD and healthy controls. The DTI-ALPS index in patients with ESRD was lower than that in healthy controls (1.460 vs. 1.632, p = 0.003). In addition, there was a significant positive correlation between the DTI-ALPS index and serum parathyroid hormone levels (r = 0.357, p = 0.011). CONCLUSION: We demonstrated glymphatic dysfunction in patients with ESRD, as revealed by the DTI-ALPS index. This study also reveals the feasibility of the DTI-ALPS method to determine glymphatic function in patients with ESRD, which could be used in future research studies.

Feasibility Study on a Segmented Ferrofluid Flow Linear Generator for Increasing the Time-Varying Magnetic Flux.

Nanoparticles and nanofluids have been implemented in energy harvesting devices, and energy harvesting based on magnetic nanofluid flow was recently achieved by using a layer-built magnet and micro-bubble injection to induce a voltage on the order of 10-1 mV. However, this is not yet suitable for some commercial purpose. In order to further increase the amount of electric voltage and current from this energy harvesting the air bubbles must be segmented in the base fluid, and the magnetic flux of the segmented flow should be materially altered over time. The focus of this research is on the development of a segmented ferrofluid flow linear generator that would scavenge electrical power from waste heat. Experiments were conducted to obtain the induced voltage, which was generated by moving a ferrofluid-filled capsule inside a multi-turn coil. Computations were then performed to explain the fundamental physical basis of the motion of the segmented flow of the ferrofluids and the air-layers.

Numerical Simulation on the Induced Voltage Across the Coil Terminal by the Segmented Flow of Ferrofluid and Air-Layer.

Nanoparticles and nanofluids have been implemented in energy harvesting devices, and energy harvesting based on magnetic nanofluid flow was recently achieved by using a layer-built magnet and microbubble injection to induce a voltage on the order of 10-1 mV. However, this is not yet suitable for some commercial purpose. The air bubbles must be segmented in the base fluid, and the magnetic flux of the ferrofluids should change over time to increase the amount of electric voltage and current from energy harvesting. In this study, we proposed a novel technique to achieve segmented flow of the ferrofluids and the air layers. This segmented ferrofluid flow linear generator can increase the magnitude of the induced voltage from the energy harvesting system. In our experiments, a ferrofluid-filled capsule produced time-dependent changes in the magnetic flux through a multi-turn coil, and the induced voltage was generated on the order of about 101 mV at a low frequency of 2 Hz. A finite element analysis was used to describe the time-dependent change of the magnetic flux through the coil according to the motion of the segmented flow of the ferrofluid and the air-layer, and the induced voltage was generated to the order of 102 mV at a high frequency of 12.5 Hz.

Interaction Model Between an Electrical Arc and a Material for Arc Discharge Synthesis of Metal Nanoparticles.

Metal nanoparticles are used in applications ranging from bio-diagnostics to catalysis due to the expectation to improve attributes or the performance of specific products or processes. The electric arc can be used to produce metal nanoparticles by evaporating the anode and forming the anode vapor. In order to synthesize the nanoparticles of the desired properties, the influence of various input parameters on the growth kinetics has to be fully understood. In this study, we presented two and three dimensional results of numerical simulation of the transferred electric arc taking into account the interaction model between an electric arc and two electrodes. It was found that the predicted temperature of the arc column with two electrodes was in good agreement with the measured data, and the main advantage of this model over our previous one was to predict the temperature distribution of the arc column with two electrodes by two- and three-dimensional computations.

Numerical Study on Alternating Current Breakdown Mechanism Between Sphere-Sphere Electrodes in Transformer Oil-Based Magnetic Nanofluids.

A numerical simulation was developed for magnetic nanoparticles in a liquid dielectric to investigate the AC breakdown voltage of the magnetic nanofluids according to the volume concentration of the magnetic nanoparticles. In prior research, we found that the dielectric breakdown voltage of the transformer oil-based magnetic nanofluids was positively or negatively affected according to the amount of magnetic nanoparticles under a testing condition of dielectric fluids, and the trajectory of the magnetic nanoparticles in a fabricated chip was visualized to verify the related phenomena via measurements and computations. In this study, a numerical simulation of magnetic nanoparticles in an insulating fluid was developed to model particle tracing for AC breakdown mechanisms happened to a sphere-sphere electrode configuration and to propose a possible mechanism regarding the change in the breakdown strength due to the behavior of the magnetic nanoparticles with different applied voltages.

Atomic-layer-deposition-assisted ZnO nanoparticles for oxide charge-trap memory thin-film transistors.

ZnO nanoparticles (NPs) with monolayer structures were prepared by atomic layer deposition (ALD) to use for a charge-trap layer (CTL) for nonvolatile memory thin-film transistors (MTFTs). The optimum ALD temperature of the NP formation was demonstrated to be 160 °C. The size and areal density of the ZnO NPs was estimated to be approximately 33 nm and 4.8 × 109 cm-2, respectively, when the number of ALD cycles was controlled to be 20. The fabricated MTFTs using a ZnO-NP CTL exhibited typical memory window properties, which are generated by charge-trap/de-trap processes, in their transfer characteristics and the width of the memory window (MW) increased from 0.6 to 18.0 V when the number of ALD cycles increased from 5 to 30. The program characteristics of the MTFT were markedly enhanced by the post-annealing process performed at 180 °C in an oxygen ambient due to the improvements in the interface and bulk qualities of the ZnO NPs. The program/erase (P/E) speed was estimated to be 10 ms at P/E voltages of -14 and 17 V. The memory margin showed no degradation with the lapse in retention time for 2 × 104 s and after the repetitive P/E operations of 7 × 103 cycles.

Genotoxicity testing of Persicariae Rhizoma (Persicaria tinctoria H. Gross) aqueous extracts.

Persicariae Rhizoma (PR) has been used as an anti-inflammatory and detoxification agent in Korea, and contains the biologically active dyes purple indirubin and blue indigo. Despite synthetic indigo showing genotoxic potential, thorough studies have not been carried out on the genotoxicity of PR. The potential genotoxicity of an aqueous extract of PR containing indigo (0.043%) and indirubin (0.009%) was evaluated using a standard battery of tests for safety assessment. The PR extract did not induce any genotoxic effects under the conditions of this study. The results of a reverse mutation assay in four Salmonella typhimurium strains and one Escherichia coli strain indicated that PR extract did not increase the frequency of revertant colonies in any strain, regardless of whether S9 mix was present or not. The PR extract also did not increase chromosomal aberrations in the presence or absence of S9 mix. Although slight signs of diarrhea were restrictedly detected in the mice treated with 2,000 mg/kg PR extract, no noteworthy changes in the frequency of micronucleated polychromatic erythrocytes were observed at doses ≤2,000 mg/kg in a bone marrow micronucleus test. These results indicate the potential safety of the PR extract, particularly if it is consumed in small amounts compared with the quantities used in the genotoxicity tests.

Comparison of serum biomarkers between patients with asthma and with chronic obstructive pulmonary disease.

OBJECTIVE: Asthma and chronic obstructive pulmonary disease (COPD) have distinct pathophysiological mechanisms but sometimes share similar clinical manifestations. Distinguishing between these diseases is important. This study compared the profiles of serum biomarkers between patients with asthma and those with COPD. METHODS: Serum levels of the chitinase like protein YKL-40, periostin, interleukin (IL)-18, and chemokine (C--C motif) ligand 18 (CCL18) were measured in asthma patients (n = 20), COPD patients (n = 16), and normal controls (n = 20). RESULTS: Serum levels of YKL-40 were higher in COPD patients [median (range), 55 (17-565) versus 208 (74-922) ng/mL, p < 0.0001], but no differences were observed between asthma and COPD patients after adjusting for age and forced expiratory volume in 1 s (FEV1). No differences in serum levels of periostin, IL-18, or CCL18 were observed between the patient groups. Total IgE and airway hypersensitivity were negatively correlated (r = -0.485, p = 0.007). CCL18 levels were related to patients' age in asthmatic patients (r = -0.562, p = 0.010). Serum levels of CCL18 and IL-18 were positively correlated in patients with COPD (r = 0.696, p = 0.003). CONCLUSIONS: No differences in the serum profiles of periostin, IL-18, or CCL18 were observed between patients with asthma and those with COPD. Serum levels of YKL-40 were not different between asthma and COPD patients after adjusting for age and FEV1. There were negative correlation between CCL18 and age in patients with asthma and positive correlation between IL-18 and CCL18 in patients with COPD.

Temperature Prediction in a Free-Burning Arc and Electrodes for Nanostructured Materials and Systems.

Temperature in a free-burning arc used for synthesis of nanoparticles and nanostructured materials is generally around 20,000 K just below the cathode, falling to about 15,000 K just above the anode, and decreasing rapidly in the radial direction. Therefore, the electrode erosion is indispensable for these atmospheric plasma systems, as well as for switching devices, due to the high heat flux transferred from high temperature arcs to electrodes, but experimental and theoretical works have not identified the characteristic phenomena because of the complex physical processes. To the previous study, we have focused on the arc self-induced fluid flow in a free-burning arc using the computational fluid dynamics (CFD) technique. At this time, our investigation is concerned with the whole region of free-burning high-intensity arcs including the tungsten cathode, the arc plasma and the anode using a unified numerical model for applying synthesis of nanoparticles and nanostructured materials practically.