Inhibiting Neutrophil Extracellular Traps Protects against Ultraviolet B-Induced Skin Damage: Effects of Hochu-ekki-to and DNase I.

UV-B radiation induces sunburn, and neutrophils are pivotal in this inflammation. In this study, we examined the potential involvement of neutrophil extracellular traps (NETs) in ultraviolet B (UVB)-induced skin inflammation, correlating the skin inflammation-mitigating effects of Hochu-ekki-to on UV-B irradiation and NETs. To elucidate NET distribution in the dorsal skin, male ICR mice, exposed to UVB irradiation, were immunohistologically analyzed to detect citrullinated histone H3 (citH3) and peptidylarginine deiminase 4 (PAD4). Reactive oxygen species (ROS) production in the bloodstream was analyzed. To establish the involvement of NET-released DNA in this inflammatory response, mice were UV-B irradiated following the intraperitoneal administration of DNase I. In vitro experiments were performed to scrutinize the impact of Hochu-ekki-to on A23187-induced NETs in neutrophil-like HL-60 cells. UV-B irradiation induced dorsal skin inflammation, coinciding with a significant increase in citH3 and PAD4 expression. Administration of DNase I attenuated UV-B-induced skin inflammation, whereas Hochu-ekki-to administration considerably suppressed the inflammation, correlating with diminished levels of citH3 and PAD4 in the dorsal skin. UV-B irradiation conspicuously augmented ROS and hydrogen peroxide (H2O2) production in the blood. Hochu-ekki-to significantly inhibited ROS and H2O2 generation. In vitro experiments demonstrated that Hochu-ekki-to notably inhibited A23187-induced NETs in differentiated neutrophil-like cells. Hence, NETs have been implicated in UV-B-induced skin inflammation, and their inhibition reduces cutaneous inflammation. Additionally, Hochu-ekki-to mitigated skin inflammation by impeding neutrophil infiltration and NETs in the dorsal skin of mice.

Fully automated coronary artery calcium quantification on electrocardiogram-gated non-contrast cardiac computed tomography using deep-learning with novel Heart-labelling method.

Aims: To develop an artificial intelligence (AI)-model which enables fully automated accurate quantification of coronary artery calcium (CAC), using deep learning (DL) on electrocardiogram (ECG)-gated non-contrast cardiac computed tomography (gated CCT) images. Methods and results: Retrospectively, 560 gated CCT images (including 60 synthetic images) performed at our institution were used to train AI-model, which can automatically divide heart region into five areas belonging to left main (LM), left anterior descending (LAD), circumflex (LCX), right coronary artery (RCA), and another. Total and vessel-specific CAC score (CACS) in each scan were manually evaluated. AI-model was trained with novel Heart-labelling method via DL according to the manual-derived results. Then, another 409 gated CCT images obtained in our institution were used for model validation. The performance of present AI-model was tested using another external cohort of 400 gated CCT images of Stanford Center for Artificial Intelligence of Medical Imaging by comparing with the ground truth. The overall accuracy of the AI-model for total CACS classification was excellent with Cohen's kappa of k = 0.89 and 0.95 (validation and test, respectively), which surpasses previous research of k = 0.89. Bland-Altman analysis showed little difference in individual total and vessel-specific CACS between AI-derived CACS and ground truth in test cohort (mean difference [95% confidence interval] were 1.5 [-42.6, 45.6], -1.5 [-100.5, 97.5], 6.6 [-60.2, 73.5], 0.96 [-59.2, 61.1], and 7.6 [-134.1, 149.2] for LM, LAD, LCX, RCA, and total CACS, respectively). Conclusion: Present Heart-labelling method provides a further improvement in fully automated, total, and vessel-specific CAC quantification on gated CCT.

3D-OSEM versus FORE + OSEM: Optimal Reconstruction Algorithm for FDG PET with a Short Acquisition Time.

Objective In this study, we investigated the optimal reconstruction algorithm in fluorodeoxyglucose (FDG) positron emission tomography (PET) with a short acquisition time. Materials and Methods In the phantom study, six spheres filled with FDG solution (sphere size: 6.23-37 mm; radioactivity ratio of spheres to background = 8:1) and placed in a National Electrical Manufacturers Association phantom were evaluated. Image acquisition time was 15 to 180 seconds, and the obtained image data were reconstructed using each of the Fourier rebinning (FORE) + ordered subsets expectation-maximization (OSEM) and 3D-OSEM algorithms. In the clinical study, mid-abdominal images of 19 patients were evaluated using regions of interest placed on areas of low, intermediate, and high radioactivity. All obtained images were investigated visually, and quantitatively using maximum standardized uptake value (SUV) and coefficient of variation (CV). Results In the phantom study, FORE + OSEM images with a short acquisition time had large CVs (poor image quality) but comparatively constant maximum SUVs. 3D-OSEM images showed comparatively constant CVs (good image quality) but significantly low maximum SUVs. The results of visual evaluation were well correlated with those of quantitative evaluation. Small spheres were obscured on 3D-OSEM images with short acquisition time, but image quality was not greatly deteriorated. The clinical and phantom studies yielded similar results. Conclusion FDG PET images with a short acquisition time reconstructed by FORE + OSEM showed poorer image quality than by 3D-OSEM. However, images obtained with a short acquisition time and reconstructed with FORE + OSEM showed clearer FDG uptake and more useful than 3D-OSEM in the light of the detection of lesions.

Neutrophil extracellular trap induction through peptidylarginine deiminase 4 activity is involved in 2,4,6-trinitrobenzenesulfonic acid-induced colitis.

Neutrophil extracellular traps (NETs) are induced in the innate immune response against infectious agents and are also implicated in the pathogenesis of various cancers and autoimmune diseases. Peptidylarginine deiminase 4 (PAD4), an enzyme that converts arginine to citrulline, is also involved in NET formation. In this study, we investigated the pathogenic effect of PAD4 on NETs in inflammatory bowel disease using a trinitrobenzene sulfonic acid (TNBS)-induced murine colitis model. PAD4-deficient (PAD4KO) mice were generated by CRISPR-Cas9-mediated genomic editing. NETs were triggered in peritoneal neutrophils obtained from wild-type mice by A23187 (a calcium ionophore), but these responses were completely abolished in the PAD4KO mice. Experimental colitis was induced in wild-type and PAD4KO mice via an intrarectal injection of TNBS. TNBS injection resulted in body weight loss, extensive colonic erosion, and ulceration in wildtype mice. However, these responses were significantly attenuated following the administration of Cl-amidine (an inhibitor of pan-PADs) and DNase I (an inhibitor of NET formation), in combination with PAD4KO in mice. TNBS-induced increases in myeloperoxidase activity, inflammatory cytokine expression, and NET formation in the colon were significantly reduced following the administration of Cl-amidine, DNase I injection, and PAD4KO. These findings suggest that NET formation contributes to the pathogenesis of TNBS-induced colitis via PAD4. Thus, PAD4 is a promising target for the treatment of inflammatory bowel disease.

Anodic Dehydrogenative Aromatization of Tetrahydrocarbazoles Leading to Carbazoles.

Anodic oxidation-promoted aromatization of 1,2,3,4-tetrahydrocarbazoles was achieved. Nitrogen-protected tetrahydrocarbazoles could be converted to the corresponding carbazoles with the use of bromide as a mediator. LiBr, an inexpensive bromide source, allowed for efficient transformation in the presence of AcOH.

Electrochemical Synthesis of Sultone Derivatives via Dehydrogenative C-O Bond Formation.

Electrochemical dehydrogenative C-O bond formation for the synthesis of sultones was achieved. In the presence of K2CO3 and H2O, constant current electrolysis of [1,1'-biphenyl]-2-sulfonyl chloride afforded an aryl-fused sultone quantitatively. Under the optimized conditions, a variety of sultone derivatives were obtained. Control experiments suggest that the electrochemical oxidation of the sulfonates generated in situ would afford sulfo radical intermediates.

Electrochemical Synthesis of Dibenzothiophenes via Intramolecular C-S Cyclization with a Halogen Mediator.

Electrochemical synthesis of dibenzothiophene derivatives was achieved. Several bis(biaryl) disulfides are efficiently converted to dibenzothiophenes by electrochemical oxidation. The use of Bu4NBr as a halogen mediator was essential, and wide varieties of dibenzothiophene derivatives were obtained in good yields.

Electrochemical hydrogenation of enones using a proton-exchange membrane reactor: selectivity and utility.

Electrochemical hydrogenation of enones using a proton-exchange membrane reactor is described. The reduction of enones proceeded smoothly under mild conditions to afford ketones or alcohols. The reaction occurred chemoselectively with the use of different cathode catalysts (Pd/C or Ir/C).

Emergence of Cooperative Impression With Self-Estimation, Thinking Time, and Concordance of Risk Sensitivity in Playing Hanabi.

The authors evaluate the extent to which a user's impression of an AI agent can be improved by giving the agent the ability of self-estimation, thinking time, and coordination of risk tendency. The authors modified the algorithm of an AI agent in the cooperative game Hanabi to have all of these traits, and investigated the change in the user's impression by playing with the user. The authors used a self-estimation task to evaluate the effect that the ability to read the intention of a user had on an impression. The authors also show thinking time of an agent influences impression for an agent. The authors also investigated the relationship between the concordance of the risk-taking tendencies of players and agents, the player's impression of agents, and the game experience. The results of the self-estimation task experiment showed that the more accurate the estimation of the agent's self, the more likely it is that the partner will perceive humanity, affinity, intelligence, and communication skills in the agent. The authors also found that an agent that changes the length of thinking time according to the priority of action gives the impression that it is smarter than an agent with a normal thinking time when the player notices the difference in thinking time or an agent that randomly changes the thinking time. The result of the experiment regarding concordance of the risk-taking tendency shows that influence player's impression toward agents. These results suggest that game agent designers can improve the player's disposition toward an agent and the game experience by adjusting the agent's self-estimation level, thinking time, and risk-taking tendency according to the player's personality and inner state during the game.

Application of an Electrochemical Microflow Reactor for Cyanosilylation: Machine Learning-Assisted Exploration of Suitable Reaction Conditions for Semi-Large-Scale Synthesis.

Cyanosilylation of carbonyl compounds provides protected cyanohydrins, which can be converted into many kinds of compounds such as amino alcohols, amides, esters, and carboxylic acids. In particular, the use of trimethylsilyl cyanide as the sole carbon source can avoid the need for more toxic inorganic cyanides. In this paper, we describe an electrochemically initiated cyanosilylation of carbonyl compounds and its application to a microflow reactor. Furthermore, to identify suitable reaction conditions, which reflect considerations beyond simply a high yield, we demonstrate machine learning-assisted optimization. Machine learning can be used to adjust the current and flow rate at the same time and identify the conditions needed to achieve the best productivity.

Synthesis of Morphinans through Anodic Aryl-Aryl Coupling.

The morphinans are an important class of structurally fascinating and physiologically important natural products as exemplified by the famous opium alkaloids of the morphine family. Although this class of secondary metabolites from the juice of the opium poppy capsule was already used for medicinal purposes thousands of years ago, chemical modifications are still being applied to the core structure today in order to achieve the most specific effect on the various receptor subtypes possible with the fewest possible side effects. The unusual architecture of the morphinan core has also proven to be a highly challenging target for total synthesis. This review highlights electrosynthetic approaches towards natural and semisynthetic morphinan alkaloids. The historical progress in applying anodic aryl-aryl couplings to the construction of the morphinan framework is described in chronological order while particular benefits and challenges concerning the electrochemical transformations are grouped together, including the influence of substitution patterns, protecting groups, and reaction conditions.

Cu-Catalyzed Dehydrogenative C-O Cyclization for the Synthesis of Furan-Fused Thienoacenes.

The first Cu-catalyzed dehydrogenative C-O cyclization for the synthesis of furan-fused thienoacenes is described. A variety of heteroacenes including a thieno[3,2-b]furan or a thieno[2,3-b]furan skeleton were synthesized by intramolecular C-H/O-H coupling. The use of a mixed solvent of N-methyl-2-pyrrolidone, ethylene glycol monomethyl ether, and toluene was essential for suppressing side reactions and efficiently promoting the reaction. Double C-O cyclization was also conducted to afford highly π-expanded furan-fused thienoacenes.

Electrosynthesis of Phosphacycles via Dehydrogenative C-P Bond Formation Using DABCO as a Mediator.

The first electrochemical synthesis of diarylphosphole oxides (DPOs) was achieved under mild conditions. The practical protocol employs commercially available and inexpensive DABCO as a hydrogen atom transfer (HAT) mediator, leading to various DPOs in moderate to good yields. This procedure can also be applied to the synthesis of six-membered phosphacycles, such as phenophosphazine derivatives. Mechanistic studies suggested that the reaction proceeds via an electro-generated phosphinyl radical.

High-Dose Vitamin C Administration Inhibits the Invasion and Proliferation of Melanoma Cells in Mice Ovary.

Several studies have been conducted to explore the anticancer effects of vitamin C (VC). However, the effect of high-dose VC administration on melanoma is still unknown. Therefore, in this study, we investigated the effects of high-dose VC (4 g/kg) on the invasion and proliferation of melanoma cells in various organs of mice. B16 melanoma cells (1 × 106 cells/100 µL) were intravenously injected into the tails of female mice, and VC solution (4 g/kg) was orally administered once a day for 14 d. On the 15th day, samples from the liver, lungs, jejunum, and ovaries were collected and analyzed for invasion and proliferation of melanoma cells. Oral VC administration decreased the number of dihydroxyphenylalanine (DOPA)-positive cells and gp100-positive melanoma cells in the ovaries and suppressed the invasion and proliferation of melanoma. Compared to melanoma-administered mice, macrophage inflammatory protein-2 levels and number of neutrophils were increased in the VC + melanoma-administered mice. Furthermore, the concentrations of VC, iron, and hydrogen peroxide, and the number of terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling (TUNEL)-positive cells were significantly increased in the ovaries of VC + melanoma-administered mice compared to those of melanoma-administered mice. These results suggest that VC can reduce the invasion and proliferation of melanoma cells in the ovaries, and neutrophils in the ovaries play an important role in achieving this melanoma-suppressive effect.

Evaluation of Dose Distribution and Normal Tissue Complication Probability of a Combined Dose of Cone-Beam Computed Tomography Imaging with Treatment in Prostate Intensity-Modulated Radiation Therapy.

PURPOSE: The purpose of this study is to evaluate the effects of cone-beam computed tomography (CBCT) on dose distribution and normal tissue complication probability (NTCP) by constructing a comprehensive dose evaluation system for prostate intensity-modulated radiation therapy (IMRT). METHODS: A system that could combine CBCT and treatment doses with MATLAB was constructed. Twenty patients treated with prostate IMRT were studied. A mean dose of 78 Gy was prescribed to the prostate region, excluding the rectal volume from the target volume, with margins of 4 mm to the dorsal side of the prostate and 7 mm to the entire circumference. CBCT and treatment doses were combined, and the dose distribution and the NTCP of the rectum and bladder were evaluated. RESULTS: The radiation dose delivered to 2% and 98% of the target volume increased by 0.90 and 0.74 Gy on average, respectively, in the half-fan mode and on average 0.76 and 0.72 Gy, respectively, in the full-fan mode. The homogeneity index remained constant. The percent volume of the rectum and bladder irradiated at each dose increased slightly, with a maximum increase of <1%. The rectal NTCP increased by approximately 0.07% from 0.46% to 0.53% with the addition of a CBCT dose, while the maximum NTCP in the bladder was approximately 0.02%. CONCLUSIONS: This study demonstrated a method to evaluate a combined dose of CBCT and a treatment dose using the constructed system. The combined dose distribution revealed increases of <1% volume in the rectal and bladder doses and approximately 0.07% in the rectal NTCP.

DNA demethylation increases NETosis.

Neutrophil extracellular traps (NETs) occur during the development of autoimmune diseases, cancer and diabetes. A novel form of cell death that is induced by NETs is called NETosis. Although these diseases are known to have an epigenetic component, epigenetic regulation of NETosis has not previously been explored. In the present study, we investigated the effects of epigenetic change, especially DNA demethylation, on NETosis in neutrophil-like cells differentiated from HL-60 cells, which were incubated for 72 h in the presence of 1.25% DMSO. DMSO-differentiated neutrophil-like cells tended to have increased methylation of genomic DNA. NETosis in the neutrophil-like cells was induced by the treatment with A23187, calcium ionophore, and increased by the addition of the DNMT inhibitor 5-azacytidine (Aza) during differentiation. Interestingly, Aza-stimulated neutrophil-like cell induced NETosis without treatment with A23187. Although reactive oxygen species (ROS), especially superoxide and hypochlorous acid, are important in NETosis induction, treatment with Aza decreased production of ROS, while mitochondria ROS scavenger tended to decrease Aza-induced NETosis. Moreover, the genomic DNA in Aza-stimulated neutrophil-like cell was demethylated, and the expression of peptidylarginine deiminase4 (PAD4) and citrullinated histone H3 (R2+R8+R17) was increased, but myeloperoxidase expression was unaffected. Additionally, PAD4 inhibition tended to decrease Aza-induced NETosis. The DNA demethylation induced by the DNMT inhibitor in neutrophil-like cells enhanced spontaneous NETosis through increasing PAD4 expression and histone citrullination. This study establishes a relationship between NETosis and epigenetics for the first time, and indicates that various diseases implicated to have an epigenetic component might be exacerbated by excessive NETosis also under epigenetic control.

Tranexamic Acid Protects Ovary and Testis Functions and Ameliorates Osteoporosis in Mice.

INTRODUCTION: In a rapidly aging society, the number of people suffering from osteoporosis keeps increasing. However, effective prevention strategies for osteoporosis are not yet currently available. OBJECTIVE: In this study, we examined the ameliorative effects of tranexamic acid on osteoporosis in 24-month-old mice. METHODS: During the study period, mice were orally administered tranexamic acid 3 times per week. RESULTS: Bone mineral density, which is a parameter of osteoporosis, was improved following tranexamic acid administration. In addition, female mice evidenced a stronger phenotypic improvement than male mice. In female mice treated with tranexamic acid, ovary abnormalities were reduced. Furthermore, the levels of transforming growth factor-ß, hyaluronic acid, CD44, reactive oxygen species, and apoptosis, as well as the number of infiltrated neutrophils and macrophages in the ovary were lower than those in the control or solvent-administered mice. In addition, 17ß-estradiol levels in blood increased when compared with the control or solvent-treated mice. In addition, administration of tranexamic acid to 24-month-old male mice decreased the level of apoptosis in the testis. However, the levels of 17ß-estradiol and testosterone in blood increased compared with the control or solvent-administered mice. CONCLUSIONS: The use of tranexamic acid had an ameliorative effect on osteoporosis, possibly by protecting ovaries and testes.

Formation of neutrophil extracellular traps in mitochondrial DNA-deficient cells.

Neutrophil extracellular trap (NET) formation plays an important role in inflammatory diseases. Although it is known that NET formation occurs via NADPH oxidase (NOX)-dependent and NOX-independent pathways, the detailed mechanism remains unknown. Therefore, in this study, we aimed to elucidate the role of mitochondria in NOX-dependent and NOX-independent NET formation. We generated mitochondrial DNA-deficient cells (ρ0 cells) by treating HL-60 cells with dideoxycytidine and differentiated them to neutrophil-like cells. These neutrophil-like ρ0 cells showed markedly reduced NOX-independent NET formation but not NOX-dependent NET formation. However, NET-associated intracellular histone citrullination was not inhibited in ρ0 cells. Furthermore, cells membrane disruption in NOX-dependent NET formation occurred in a Myeloperoxidase (MPO) and mixed lineage kinase domain like pseudokinase (MLKL)-dependent manner; however, cell membrane disruption in NOX-independent NET formation partially occurred in an MLKL-dependent manner. These results highlight the importance of mitochondria in NOX-independent NET formation.