NKG2D (Natural Killer Group 2, Member D) ligand expression and ameloblastoma recurrence: a retrospective immunohistological pilot study.

BACKGROUND/PURPOSE: This retrospective immunohistological pilot study aimed to investigate the influence of natural killer group 2, member D (NKG2D) ligand expression on ameloblastoma recurrence after surgical resection. It also aimed to elucidate additional clinical factors that could serve as predictors of ameloblastoma recurrence. MATERIALS AND METHODS: This study included 96 patients who were histologically diagnosed with ameloblastoma after surgical resection. The expression of NKG2D ligands, including UL16-binding proteins (ULBPs) 1-3 and major histocompatibility complex class I chain-related molecule (MIC) A/B, was evaluated in formalin-fixed paraffin-embedded tumor tissues via immunohistochemistry assays. Furthermore, the patients' electronic medical records were reviewed. Multivariate Cox regression analysis was conducted, and data were expressed as adjusted hazard ratios [HRs] with 95% confidence intervals [95% CIs]. RESULTS: Multivariate analysis revealed that recurrent tumors (ref.: primary; adjusted HR [95% CI]: 2.780 [1.136, 6.803], p = 0.025) and positive MICA/B expression (ref.: negative; adjusted HR [95% CI]: 0.223 [0.050, 0.989], p = 0.048) independently affected recurrence-free survival in ameloblastoma. CONCLUSION: This study identified recurrent cases and loss of MICA/B expression as independent predictors of early ameloblastoma recurrence following surgical resection. The findings suggest that decreased MICA/B expression might undermine NKG2D-mediated tumor immunosurveillance, thereby influencing early recurrence.

Unraveling the Intrinsic Thermal Behavior of Freestanding Ionomer Films Depending on Thickness from Commercial Membrane to Nanofilm.

Proton exchange membrane fuel cells (PEMFCs) for automotive applications are required to achieve mechanical reliability at various temperatures ranging from subfreezing to 80 °C. The thermal behavior of the electrode should be considered at the initial design stage to design a robust automotive fuel cell electrode. Recently, a behavior different from that of the bulk state has been reported for ionomers with a few nanometers of thickness. Therefore, the intrinsic thermal behavior of ionomer films with thicknesses from micrometers to nanometers is quantitatively investigated in this study. By introducing the fabrication of a pseudo-freestanding Nafion thin film and in-plane thermal strain measurement method on the water surface, the thermal expansion of the freestanding Nafion thin film was successfully measured with minimizing substrate constraints. Thermal strain measurement and X-ray scattering studies revealed that the weakening of intermolecular interaction within the hydrophobic and hydrophilic domains in the Nafion thin film caused thermal expansion, and well-structured hydrophobic domains could suppress thermal expansion. The thermal expansion behavior with different heat treatments provides evidence of the thin-film-to-bulk transition of the fully hydrated Nafion film. Intrinsic thermal behavior without substrate interactions can facilitate an understanding of the thermal behavior of electrodes and provide insight into designing a robust PEMFC in temperature-varying environments.

Distribution and elimination kinetics of midazolam and metabolites after post-resuscitation care: a prospective observational study.

Administration of sedatives for post-resuscitation care can complicate the determination of the optimal timing to avoid inappropriate, pessimistic prognostications. This prospective study aimed to investigate the distribution and elimination kinetics of midazolam (MDZ) and its metabolites, and their association with awakening time. The concentrations of MDZ and its seven metabolites were measured immediately and at 4, 8, 12, and 24 h after the discontinuation of MDZ infusion, using liquid chromatography-tandem mass spectrometry. The area under the time-plasma concentration curve from 0 to 24 h after MDZ discontinuation (AUClast) was calculated based on the trapezoidal rule. Of the 15 enrolled patients, seven awakened after the discontinuation of MDZ infusion. MDZ and three of its metabolites were major compounds and their elimination kinetics followed a first-order elimination profile. In the multivariable analysis, only MDZ was associated with awakening time (AUClast: R2 = 0.59, p = 0.03; AUCinf: R2 = 0.96, p < 0.001). Specifically, a 0.001% increase in MDZ AUC was associated with a 1% increase in awakening time. In the individual regression analysis between MDZ concentration and awakening time, the mean MDZ concentration at awakening time was 16.8 ng/mL. The AUC of MDZ is the only significant factor associated with the awakening time.

Impaired Capillary Recruitment Capacity in Obesity: A Subgroup Analysis of Prospective Observational Study on Anesthesia Effects.

BACKGROUND This subgroup analysis of prospective observational research, involving 71 participants, compared the effects of anesthesia on microvascular reactivity in obese vs lean individuals using near-infrared spectroscopy and vascular occlusion tests. The correlation between the body mass index (BMI) and microvascular reactivity under general anesthesia was also investigated. MATERIAL AND METHODS This study enrolled adult patients classified as American Society of Anesthesiologists physical status I or II, undergoing elective surgery under general anesthesia. The microcirculatory variables measured before (Tpre) and 30 min following the induction of anesthesia (Tpost) were as follows: baseline tissue oxygen saturation (StO2), occlusion slope (∇occl), and recovery slope (∇recov). The patients were grouped according to their BMI (lean [BMI <25 kg/m²] vs obese [BMI ≥25 kg/m²]). Data are presented as medians and interquartile ranges. RESULTS There were 43 patients in the lean group and 28 in the obese group. At Tpre, baseline StO2, ∇occl, and ∇recov were not different between the 2 groups (P=0.860, 0.659, and 0.518, respectively). At Tpost, the baseline StO2 and ∇occl were not different between the 2 groups (P=0.343 and 0.791); however, the ∇recov was lower in the obese group than in the lean group (3.245 [2.737, 3.977] vs 4.131 [3.491, 4.843], P=0.003). At Tpost, BMI showed a moderate correlation with ∇recov (correlation coefficient: -0.319, P=0.007). CONCLUSIONS In obese patients, capillary recruitment capacity during general anesthesia is compromised compared to lean patients.

Protective Effects of Lanostane Triterpenoids from Chaga Mushroom in Human Keratinocytes, HaCaT Cells, against Inflammatory and Oxidative Stresses.

Inotodiol, a lanostane-type triterpenoid, and many phytochemicals from Chaga mushrooms have been investigated for various allergic diseases. However, the anti-aging and anti-inflammatory activities of inotodiol under different types of oxidative stress and the impact of inotodiol on collagen and hyaluronan synthesis have not been sufficiently studied. Lanostane triterpenoids-rich concentrate, which contained 10% inotodiol as major (inotodiol concentrate), was prepared from Chaga and compared with pure inotodiol in terms of anti-inflammatory activities on a human keratinocyte cell line, HaCaT cells, under various stimulations such as stimulation with ultraviolet (UV) B or tumor necrosis factor (TNF)-α. In stimulation with TNF-α, interleukin (IL)-1ß, IL-6, and IL-8 genes were significantly repressed by 0.44~4.0 µg/mL of pure inotodiol. UVB irradiation induced the overexpression of pro-inflammatory cytokines, but those genes were significantly suppressed by pure inotodiol or inotodiol concentrate. Moreover, pure inotodiol/inotodiol concentrate could also modulate the synthesis of collagen and hyaluronic acid by controlling COL1A2 and HAS2/3 expression, which implies a crucial role for pure inotodiol/inotodiol concentrate in the prevention of skin aging. These results illuminate the anti-inflammatory and anti-aging effects of pure inotodiol/inotodiol concentrate, and it is highly conceivable that pure inotodiol and inotodiol concentrate could be promising natural bioactive substances to be incorporated in therapeutic and beautifying applications.

Isolation, Physicochemical Characterization, and Biological Properties of Inotodiol, the Potent Pharmaceutical Oxysterol from Chaga Mushroom.

Inotodiol, an oxysterol found only in Chaga mushroom, has received attention from the pharmaceutical industry due to its strong antioxidant and anti-allergic activities. However, the production of inotodiol is still challenging, and its fundamental properties have yet to be investigated. This study aims to develop an efficient method to produce high-purity inotodiol from Chaga mushroom. Then, pure inotodiol was used to assess its physicochemical properties and biological activities. By optimizing the solvent used for extraction and purification, a new method to produce inotodiol was developed with high purity (>97%) and purification yield (33.6%). Inotodiol exhibited a melting point (192.06 °C) much higher than lanosterol and cholesterol. However, the solubility of inotodiol in organic solvents was notably lower than those of the other two sterols. The difference in the hydroxyl group at C-22 of inotodiol has shown the distinctive physicochemical properties of inotodiol compared with cholesterol and lanosterol. Based on those findings, a nonionic surfactant-based delivery system for inotodiol was developed to improve its bioavailability. The inotodiol microemulsion prepared with 1-2% Tween-80 exhibited homogenous droplets with an acceptable diameter (354 to 217 nm) and encapsulation efficiency (85.6-86.9%). The pharmacokinetic analysis of inotodiol microemulsion in oral administration of 4.5 mg/kg exhibited AUC0-24h = 341.81 (ng·h/mL), and Cmax = 88.05 (ng/mL). Notably, when the dose increased from 4.5 to 8.0 mg/kg, the bioavailability of inotodiol decreased from 41.32% to 33.28%. In a mouse model of sepsis, the serum level of interleukin-6 significantly decreased, and the rectal temperature of mice was recovered in the inotodiol emulsion group, indicating that inotodiol microemulsion is an effective oral delivery method. These results could provide valuable information for applying inotodiol in functional food, cosmetic, and pharmaceutical industries.

Dynamic stability of cholesterol and desmosterol in human milk from four Asian countries.

Cholesterol is one of the functional nutrients in human milk, which is indispensable for infant growth. In this study, the concentration of cholesterol and desmosterol in human milk from four Asian countries (n = 578), including Korea, China, Vietnam, and Pakistan, were investigated. The average cholesterol concentrations of Korea and China were similar ranging between 90.2-91.6 mg/L, but those from Vietnam and Pakistan were higher at 113.8 and 175.7 mg/L, respectively. The relative standard deviations were 31-36%, except for Pakistan (51%), showing a broad distribution of 48 to 612 mg/L. Desmosterol concentrations were similar, ranging between 11.2 and 12.8 mg/L except for Pakistan, which was lower than other countries at 9.4 mg/L. In addition, the cholesterol and desmosterol concentrations during the lactation periods were not significantly different in all four Asian countries. Mothers' BMI did not significantly impact the cholesterol and desmosterol concentration in maternal milk within the same country. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01141-9.

The analysis of 2'-fucosyllactose concentration in Korean maternal milk using LC-MS/MS.

Despite health benefits reported recently, 2'-fucosyllactose (2'-FL) concentration in maternal milk was not conclusively reported because it varies between countries and mothers. Particularly, its distribution among Korean mothers was not obtained from a reliable sample group yet. Thus, a dynamic range for 2'-FL concentration in Korean mothers' milk was investigated from 102 samples. A quantitative method using multiple reaction monitoring (MRM) by triple-quadrupole-mass spectrometry has been evaluated by a standard procedure of method validation. The 2'-FL concentration was in the range of 0.4 to 2.6 g/L overall. While the samples from secretor mothers (n = 80) contained 1.0 to 2.8 g/L of 2'-FL, the maternal milk from non-secretor mothers (n = 22) had 0.01 to 0.06 g/L of 2'-FL only. In addition to the genetic variation of mothers, the lactation period impacted the 2'-FL concentration. The average 2'-FL concentration of the late-stage group (> 60 days) was 78% of that obtained from the first month of postpartum mothers. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01154-4.

Evaluation of Toxicity and Efficacy of Inotodiol as an Anti-Inflammatory Agent Using Animal Model.

Chaga mushroom (Inonotus obliquus) comprises polyphenolic compounds, triterpenoids, polysaccharides, and sterols. Among the triterpenoid components, inotodiol has been broadly examined because of its various biological activities. The purpose of this study is to examine inotodiol from a safety point of view and to present the potential possibilities of inotodiol for medical usage. From chaga mushroom extract, crude inotodiol (INO20) and pure inotodiol (INO95) were produced. Mice were treated with either INO20 or INO95 once daily using oral administration for repeated dose toxicity evaluation. Serum biochemistry parameters were analyzed, and the level of pro-inflammatory cytokines in the serum was quantified. In parallel, the effect of inotodiol on food allergic symptoms was investigated. Repeated administration of inotodiol did not show any mortality or abnormalities in organs. In food allergy studies, the symptoms of diarrhea were ameliorated by administration with INO95 and INO20. Furthermore, the level of MCPT-1 decreased by treatment with inotodiol. In this study, we demonstrated for the first time that inotodiol does not cause any detrimental effect by showing anti-allergic activities in vivo by inhibiting mast cell function. Our data highlight the potential to use inotodiol as an immune modulator for diseases related to inflammation.

Activation of galactose utilization by the addition of glucose for the fermentation of agar hydrolysate using Lactobacillus brevis ATCC 14869.

OBJECTIVE: To investigate the application of carbon catabolite repression (CCR) relaxed Lactobacillus brevis ATCC 14869 in the utilization of agar hydrolysate to produce bioethanol and lactic acid through fermentation. RESULTS: As a single carbon source, galactose was not metabolized by L. brevis. However, L. brevis consumed galactose simultaneous to glucose and ceased cell growth after depletion of glucose. For complete use of galactose from agar hydrolysis, glucose need to be periodically replenished into the growth medium. Overall, L. brevis successfully used agar hydrolysate and produced 17.2 g/L of ethanol and 31.9 g/L of lactic acid. The maximum specific cell growth rate on galactose and glucose mixture was the same with the glucose-only medium at 0.12 h-1. The molar product yields from glucose for lactic acid and ethanol were 1.02 and 0.95 respectively, equal to values obtained from the simultaneous utilization of glucose and galactose. CONCLUSION: In contribution to the ongoing efforts to utilize marine biomass, the relaxed CCR in Lactobacillus brevis ATCC 14869 was herein exploited to produce bioethanol and lactic acid from red seaweed hydrolysates.

Isomer-Specific Monitoring of Sialylated N-Glycans Reveals Association of α2,3-Linked Sialic Acid Epitope With Behcet's Disease.

Behcet's disease (BD) is an immune disease characterized by chronic and relapsing systemic vasculitis of unknown etiology, which can lead to blindness and even death. Despite continuous efforts to discover biomarkers for accurate and rapid diagnosis and optimal treatment of BD, there is still no signature marker with high sensitivity and high specificity. As the link between glycosylation and the immune system has been revealed, research on the immunological function of glycans is being actively conducted. In particular, sialic acids at the terminus of glycoconjugates are directly implicated in immune responses, cell-cell/pathogen interactions, and tumor progression. Therefore, changes in sialic acid epitope in the human body are spotlighted as a new indicator to monitor the onset and progression of immune diseases. Here, we performed global profiling of N-glycan compositions derived from the sera of 47 healthy donors and 47 BD patients using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) to preferentially determine BD target glycans. Then, three sialylated biantennary N-glycans were further subjected to the separation of linkage isomers and quantification using porous graphitized carbon-liquid chromatography (PGC-LC)/multiple reaction monitoring (MRM)-MS. We were able to successfully identify 11 isomers with sialic acid epitopes from the three glycan compositions consisting of Hex5HexNAc4NeuAc1, Hex5HexNAc4Fuc1NeuAc1, and Hex5HexNAc4NeuAc2. Among them, three isomers almost completely distinguished BD from control with high sensitivity and specificity with an area under the curve (AUC) of 0.945, suggesting the potential as novel BD biomarkers. In particular, it was confirmed that α2,3-sialic acid at the terminus of biantennary N-glycan was the epitope associated with BD. In this study, we present a novel approach to elucidating the association between BD and glycosylation by tracing isomeric structures containing sialic acid epitopes. Isomer-specific glycan profiling is suitable for analysis of large clinical cohorts and may facilitate the introduction of diagnostic assays for other immune diseases.

Variation of Triterpenic Acids in 12 Wild Syzygium formosum and Anti-Inflammation Activity on Human Keratinocyte HaCaT.

Syzygium formosum (Wall.) Masam leaf is known as a Vietnamese traditional herbal medicine used to treat atopic dermatitis and stomach ulcers. Recently, its potent anti-allergic effects were reported with possible active compounds analysis. Here, we collected S. formosum leaves from 12 wild trees and compared compositions of triterpenic acids (TA) with Centella asiatica. Anti-inflammatory activities of S. formosum leaf extract (SFLE) was compared with C. asiatica extract (CAE) using human keratinocyte, HaCaT. In this study, up to seven TAs were identified in SFLE, while only madecassic and asiatic acids were detected in the CAE. Total TA content varied among SFLE, but asiatic, corosolic, and betulinic acids were the major components. Surprisingly, wild tree sample 12 (S12) contained total TA of 27.2 mg/g dry-leaves that was 5-fold greater than that in the C. asiatica sample, and S4 had the highest content of asiatic acid (12.6 mg/g dry-leaves) that accounted for 50% of the total TA. S4 and S12 showed more than 3-fold higher anti-oxidative power than the CAE. In the UVB irradiation model, S4 and S12 (5 µg/mL) strongly repressed mRNA levels of pro-inflammatory cytokines (IL-1ß, IL-6, and IL-8) and COX-2, while the CAE at the same condition showed moderate or weak repression. The difference in anti-inflammation effects between the SFLE and the CAE was also confirmed by protein quantifications. Taken together, SFLE has great potentials as a new cosmeceutical ingredient with a higher amount of skin-active phytochemicals.

Detection of Aberrant Glycosylation of Serum Haptoglobin for Gastric Cancer Diagnosis Using a Middle-Up-Down Glycoproteome Platform.

Gastric cancer is a frequently occurring cancer and is the leading cause of cancer-related deaths. Recent studies have shown that aberrant glycosylation of serum haptoglobin is closely related to gastric cancer and has enormous potential for use in diagnosis. However, there is no platform with high reliability and high reproducibility to comprehensively analyze haptoglobin glycosylation covering microheterogeneity to macroheterogeneity for clinical applications. In this study, we developed a middle-up-down glycoproteome platform for fast and accurate monitoring of haptoglobin glycosylation. This platform utilizes an online purification of LC for sample desalting, and an in silico haptoglobin glycopeptide library constructed by combining peptides and N-glycans to readily identify glycopeptides. In addition, site-specific glycosylation with glycan heterogeneity can be obtained through only a single MS analysis. Haptoglobin glycosylation in clinical samples consisting of healthy controls (n = 47) and gastric cancer patients (n = 43) was extensively investigated using three groups of tryptic glycopeptides: GP1 (including Asn184), GP2 (including Asn207 and Asn211), and GP3 (including Asn241). A total of 23 individual glycopeptides were determined as potential biomarkers (p < 0.00001). In addition, to improve diagnostic efficacy, we derived representative group biomarkers with high AUC values (0.929 to 0.977) through logistic regression analysis for each GP group. It has been found that glycosylation of haptoglobin is highly associated with gastric cancer, especially the glycosite Asn241. Our assay not only allows to quickly and easily obtain information on glycosylation heterogeneity of a target glycoprotein but also makes it an efficient tool for biomarker discovery and clinical diagnosis.

In-depth characterization of non-human sialic acid (Neu5Gc) in human serum using label-free ZIC-HILIC/MRM-MS.

Sialic acid Neu5Gc, a non-human glycan, is recognized as a new harmful substance that can cause vascular disease and cancer. Humans are unable to synthesize Neu5Gc due to a genetic defect that converts Neu5Ac to Neu5Gc, but Neu5Gc is often observed in human biological samples. Therefore, the demand for accurately measuring the amount of Neu5Gc present in human blood or tissues is rapidly increasing, but there is still no method to reliably quantify trace amounts of a non-human sugar. In particular, selective isolation and detection of Neu5Gc from human serum is analytically challenging due to the presence of excess sialic acid Neu5Ac, which has physicochemical properties very similar to Neu5Gc. Herein, we developed the label-free approach based on ZIC-HILIC/MRM-MS that can enrich sialic acids released from human serum and simultaneously monitor Neu5Ac and Neu5Gc. The combination of complete separation of Neu5Gc from abundant Neu5Ac by hydrophilic and electrostatic interactions with selective monitoring of structure-specific cross-ring cleavage ions generated by negative CID-MS/MS was remarkably effective for quantification of Neu5Ac and Neu5Gc at the femtomole level. Indeed, we were able to successfully determine the absolute quantitation of Neu5Gc from 30 healthy donors in the range of 3.336 ± 1.252 pg/µL (mean ± SD), 10,000 times lower than Neu5Ac. In particular, analysis of sialic acids in protein-free serum revealed that both Neu5Ac and Neu5G are mostly bound to proteins and/or lipids, but not in free form. In addition, the correlation between expression level of Neu5Gc and biological factors such as BMI, age, and sex was investigated. This method can be widely used in studies requiring sialic acid-related measurements such as disease diagnosis or prediction of immunogenicity in biopharmaceuticals as it is both fast and highly sensitive.

A novel maltoheptaose-based sugar ester having excellent emulsifying properties and optimization of its lipase-catalyzed synthesis.

A novel maltoheptaose-palmitate ester (G7-PA) was synthesized and investigated for emulsion properties. First of all, the optimal conditions for lipase-catalyzed G7-PA synthesis, which were 0.2 of the G7/PA molar ratio, 33.5 U of immobilized CALB per 1 g of PA in 10% DMSO, were determined by response surface methodology. G7-PA was compared with the commercial sucrose-PA (S-PA) in terms of emulsion-forming ability and stability at extreme conditions. At the 0.1% surfactant concentration, G7-PA emulsion exhibited a droplet distribution similar to the 0.2% surfactant condition, while S-PA emulsion was quickly destabilized. G7-PA showed better emulsifying properties than the S-PA at the acidic condition (pH 3). Flocculation and phase separation was observed at the S-PA emulsion, but the G7-PA emulsion was stable for 7-day. In thermostability tests, G7-PA and S-PA both were stable up to the boiling temperature. Conclusively, G7-PA exhibits excellent properties as a biosurfactant in O/W emulsion compared with S-PA.

Comprehensive analysis of fatty acids in human milk of four Asian countries.

Human milk lipids provide not only energy but also indispensable bioactive components such as essential fatty acids. To establish the recommended daily intake value and guidelines for infant formula, a reference library of fatty acid composition has been generated from 4 Asian countries (South Korea, China, Vietnam, and Pakistan). Regardless of country, palmitic acid (C16:0), linoleic acid (C18:1), and linolenic acid (C18:2) were the 3 most abundant fatty acids in human milk and account for more than 75% of total fatty acids (total FA). However, there were several considerable differences between fatty acids, particularly n-3 and n-6 (omega-3 and omega-6) groups. Chinese mothers' milk had a high concentration of linoleic acid at 24.38 ± 10.02% of total FA, which may be due to maternal diet. Among the 4 countries, Pakistani mothers' milk contained a high amount of saturated fatty acid (56.83 ± 5.96% of total FA), and consequently, polyunsaturated fatty acids, including n-3 and n-6, were significantly lower than in other countries. It is noteworthy that docosahexaenoic acid (DHA) in Pakistani mothers' milk was 44.8 ± 33.3 mg/L, which is only 25 to 30% of the levels in the other 3 countries, suggesting the need for DHA supplementation for infants in Pakistan. Moreover, the ratio of n-6 to n-3 was also remarkably high in Pakistani mothers' milk (15.21 ± 4.96), being 1.4- to 1.7-fold higher than in other countries. The average DHA:ARA ratio in Asian human milk was 1.01 ± 0.79. Korean mothers' milk showed a high DHA:ARA ratio, with a value of 1.30 ± 0.98, but Pakistani mothers' milk had a significantly lower value (0.42 ± 0.12). The fatty acid compositions and anthropometric data of mother (body mass index, age) did not show any correlation. The obtained data might provide information about human milk compositions in the Asian region that could benefit from setting up recommended nutrient intake and infant formula for Asian babies.

Comparison of Catalyzing Properties of Bacterial 4-α-Glucanotransferases Focusing on Their Cyclizing Activity.

A newly cloned 4-α-glucanotransferase (αGT) from Deinococcus geothermalis and two typical bacterial αGTs from Thermus scotoductus and Escherichia coli (MalQ) were investigated. Among 4 types of catalysis, the cyclization activity of αGTs that produces cycloamylose (CA), a valuable carbohydrate making inclusion complexes, was intensively studied. The new αGT, DgαGT, showed close protein sequence to the αGT from T. scotoductus (TsαGT). MalQ was clearly separated from the other two αGTs in the phylogenetic and the conserved regions analyses. The reaction velocities of disproportionation, cyclization, coupling, and hydrolysis of three αGTs were determined. Intriguingly, MalQ exhibited more than 100-fold lower cyclization activity than the others. To lesser extent, the disproportionation activity of MalQ was relatively low. DgαGT and TsαGT showed similar kinetics results, but TsαGT had nearly 10-fold lower hydrolysis activity than DgαGT. Due to the very low cyclizing activity of MalQ, DgαGT and TsαGT were selected for further analyses. When amylose was treated with DgαGT or TsαGT, CA with a broad DP range was generated immediately. The DP distribution of CA had a bimodal shape (DP 7 and 27 as peaks) for the both enzymes, but larger DPs of CA quickly decreased in the DgαGT. Cyclomaltopentaose, a rare cyclic sugar, was produced at early reaction stage and accumulated as the reactions went on in the both enzymes, but the increase was more profound in the TsαGT. Taken together, we clearly demonstrated the catalytic differences between αGT groups from thermophilic and pathogenic bacteria that and showed that αGTs play different roles depending on their lifestyle.

Spatial and temporal diversity of glycome expression in mammalian brain.

Mammalian brain glycome remains a relatively poorly understood area compared to other large-scale "omics" studies, such as genomics and transcriptomics due to the inherent complexity and heterogeneity of glycan structure and properties. Here, we first performed spatial and temporal analysis of glycome expression patterns in the mammalian brain using a cutting-edge experimental tool based on liquid chromatography-mass spectrometry, with the ultimate aim to yield valuable implications on molecular events regarding brain functions and development. We observed an apparent diversity in the glycome expression patterns, which is spatially well-preserved among nine different brain regions in mouse. Next, we explored whether the glycome expression pattern changes temporally during postnatal brain development by examining the prefrontal cortex (PFC) at different time point across six postnatal stages in mouse. We found that glycan expression profiles were dynamically regulated during postnatal developments. A similar result was obtained in PFC samples from humans ranging in age from 39 d to 49 y. Novel glycans unique to the brain were also identified. Interestingly, changes primarily attributed to sialylated and fucosylated glycans were extensively observed during PFC development. Finally, based on the vast heterogeneity of glycans, we constructed a core glyco-synthesis map to delineate the glycosylation pathway responsible for the glycan diversity during the PFC development. Our findings reveal high levels of diversity in a glycosylation program underlying brain region specificity and age dependency, and may lead to new studies exploring the role of glycans in spatiotemporally diverse brain functions.

Discovery of N-glycan Biomarkers for the Canine Osteoarthritis.

Protein glycosylation is a post-translational modification that impacts on protein activity, stability, and interactions. It was sensitively altered by the cellular state and, therefore, is now used for a diagnostic or prognostic indicator of various human diseases such as cancer. To evaluate the clinical feasibility in the veterinary area, the N-glycan biomarkers were discovered from canine serum for the diagnosis of osteoarthritis (OA), which is one of the most common diseases of dogs. N-glycome was obtained from 20 µL of canine serum by the enzymatic cleavage followed by the purification and enrichment using solid-phase extraction. Independent compositions of 163 and 463 N-glycans were found from healthy control (n = 41) and osteoarthritis patients (n = 92), respectively. Initially, 31 of the potential biomarkers were screened by the p-values below 1.0 × 10-10 from ANOVA. Then, the area under the curve (AUC) and the intensity ratio between OA patient and healthy control (P/C ratio) were calculated. Considering the diagnostic efficacy, the AUC bigger than 0.9 and the P/C ratio larger than 3.0 were used to discover 16 N-glycans as diagnostic biomarkers. Particularly, five of the diagnostic biomarkers were AUC above 0.99 and three of N-glycans had AUC 1.0. The results suggest a clear possibility for N-glycan biomarkers to be used as a clinical tool in the veterinary medical area enabling to provide objective and non-invasive diagnostic information.

Development of Real-Time Hand Gesture Recognition for Tabletop Holographic Display Interaction Using Azure Kinect.

The use of human gesturing to interact with devices such as computers or smartphones has presented several problems. This form of interaction relies on gesture interaction technology such as Leap Motion from Leap Motion, Inc, which enables humans to use hand gestures to interact with a computer. The technology has excellent hand detection performance, and even allows simple games to be played using gestures. Another example is the contactless use of a smartphone to take a photograph by simply folding and opening the palm. Research on interaction with other devices via hand gestures is in progress. Similarly, studies on the creation of a hologram display from objects that actually exist are also underway. We propose a hand gesture recognition system that can control the Tabletop holographic display based on an actual object. The depth image obtained using the latest Time-of-Flight based depth camera Azure Kinect is used to obtain information about the hand and hand joints by using the deep-learning model CrossInfoNet. Using this information, we developed a real time system that defines and recognizes gestures indicating left, right, up, and down basic rotation, and zoom in, zoom out, and continuous rotation to the left and right.