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Abstract Background Malignant gliomas exhibit rapid tumor progression and resistance to treatment, leading to high lethality. One of the causes is the reduced progression of apoptosis in glioma cells. Layilin is a type 1 transmembrane protein with a C-type lectin motif in its extracellular domain. We previously reported that layilin is mainly localized to mitochondria or their close proximity and that layilin is essential for maintaining of the fragmented type of mitochondria. This study investigates the effects of layilin on mitochondria-mediated apoptosis, focusing on B cell lymphoma (BCL)-2 family proteins in a glioma cell line of A172 cells. Results We compared the levels of pro-apoptotic BCL-2 family proteins of BAD, BAK, BAX, and BIM and anti-apoptotic BCL-2 family proteins of BCL-2 and BCL-XL between layilin- knockdown (KD) cells and control cells using western blot. The protein levels of BAD were significantly smaller in layilin-KD cells than in control cells, while those of BCL-2 were significantly larger. We then compared the mitochondrial membrane potential (ΔΨm) under p-trifluoromethoxyphenyl hydrazone (FCCP)-treated conditions using MT-1 staining. In layilin-KD cells, ΔΨm was significantly larger and FCCP-induced ΔΨm reduction was significantly lower than in control cells. Furthermore, we examined the levels of cell membrane-bound Annexin V and DNA-bound propidium idodide (PI) in layilin-KD cells with/without staurosporine (STS) treatment. Layilin-KD significantly decreased levels of cell membrane-bound Annexin V with/without STS treatment. On the other hand, PI levels were not changed by layilin-KD. We also investigated the amounts of the active caspase (CASP)-3, CASP-6, CASP-7, and poly (ADP-ribose) polymerase-1 (PARP1, cleaved form), as well as DNA fragmentation in layilin-KD cells under apoptotic conditions induced by STS, using western blot and the DNA ladder method, respectively. Under STS-treated conditions, the amounts of active CASP-3, CASP-7, and poly (ADP-ribose) PARP1 were significantly smaller in layilin-KD cells than in control cells. Accordingly, DNA fragmentation was significantly suppressed in layilin-KD cells compared to control cells under STS-treated conditions. Conclusion This study demonstrates that layilin contributes to ΔΨm reduction to promote apoptosis by up-regulating BAD and down-regulating BCL-2 in glioma cells. Our data elucidates a new function of layilin: regulation of mitochondria-mediated apoptosis.

Introduction: Finegoldia magna is a member of the Gram-positive anaerobic cocci group and constitutes the flora of the skin and other parts of the body. It sometimes colonizes diabetic foot and rarely infects skin or soft tissue of non-immunocompromised patients. Case Presentation: Here, we report the case of a severe subcutaneous abscess on the back caused by F. magna involving an immunocompromised patient with poorly controlled diabetes. A 48-year-old woman with diabetes mellitus and anemia associated with uterine fibroids was referred to us with a 1-month history of a skin manifestation on her back, with a body temperature of 35.9°C and blood pressure of 115/73 mm Hg. The manifestation involved a subcutaneous mass of 36 × 45 cm with a foul odor, partly covered with necrotic tissue, which had the appearance of a tortoiseshell-like pattern. Blood examination revealed C-reactive protein of 21.4 mg/dL and hemoglobin A1c of 9.1%. Contrast-enhanced computed tomography showed a subcutaneous abscess with internal emphysema. Emergency debridement was performed, resulting in drainage of foul-smelling gray-green pus. F. magna was detected in the pus and skin tissue. Conclusion: Skin and soft tissue infectious disease caused by F. magna is extremely rare, but the disease tends to become severe once developing in an immunocompromised patient, such as a patient with poorly controlled diabetes. Therefore, physicians should consider F. magna as a causative agent when poorly controlled diabetic patients suffer from severe infectious cutaneous manifestations.

Abstract Plasma β-amyloid (Aβ) assays are a promising tool for Alzheimer’s disease diagnosis in clinical practice. To obtain reliable results, establishing an appropriate sample-handling procedure for each analytical platform is warranted. This study proposes an appropriate sample-handling procedure using HISCL analyzer by elucidating the individual/combined effects of pre-analytical parameters on plasma Aβ42/Aβ40 levels. We investigated the effects of various pre-analytical parameters, including storage times for whole blood, plasma, and freezing conditions, on plasma Aβ42/Aβ40 levels, and confirmed if these values met the acceptable criteria. Plasma Aβ42/Aβ40 levels were acceptable in all conditions. We determined our protocol by confirming that plasma Aβ42/Aβ40 levels remained acceptable when combining pre-analytical parameters. We established an appropriate sample-handling protocol that ensures reliable measurement of plasma Aβ42/Aβ40 levels using HISCL analyzer. We believe the Aβ assay, with our protocol, shows promise for aiding AD diagnosis in clinical settings.

ABSTRACT Purpose: Amid rising health awareness, natural products which has milder effects than medical drugs are becoming popular. However, only few systems can quantitatively assess their impact on living organisms. Therefore, we developed a deep-learning system to automate the counting of cells in a gerbil model, aiming to assess a natural product’s effectiveness against ischemia. Methods: The image acquired from paraffin blocks containing gerbil brains was analyzed by a deep-learning model (fine-tuned Detectron2). Results: The counting system achieved a 79%-positive predictive value and 85%-sensitivity when visual judgment by an expert was used as ground truth. Conclusions: Our system evaluated hydrogen water’s potential against ischemia and found it potentially useful, which is consistent with expert assessment. Due to natural product’s milder effects, large data sets are needed for evaluation, making manual measurement labor-intensive. Hence, our system offers a promising new approach for evaluating natural products.

IntroductionThe clinical relevance of soluble forms of programmed cell death-1 (sPD-1) and programmed cell death-ligand 1 (sPD-L1) remains unclear. We here investigated the relation between the efficacy of PD-1 blockade and pretreatment plasma levels of sPD-1 and sPD-L1 across a broad range of cancer types.MethodsWe retrospectively analyzed clinical data from 171 patients with advanced solid tumors who received nivolumab or pembrolizumab monotherapy regardless of treatment line. The concentrations of sPD-1 and sPD-L1 were measured with a fully automated immunoassay (HISCL system).ResultsThe study subjects comprised patients with head and neck cancer (n = 50), urothelial cancer (n = 42), renal cell cancer (n = 37), gastric cancer (n = 20), esophageal cancer (n = 10), malignant pleural mesothelioma (n = 6), or microsatellite instability-high tumors (n = 6). High or low levels of sPD-1 or sPD-L1 were not significantly associated with progression-free survival (PFS) or overall survival (OS) for PD-1 blockade in the entire study population. Comparison of treatment outcomes according to combinations of high or low sPD-1 and sPD-L1 levels, however, revealed that patients with low sPD-1 and high sPD-L1 concentrations had a significantly poorer PFS (HR of 1.79 [95% CI, 1.13–2.83], p = 0.01) and a tendency toward poorer OS (HR of 1.70 [95% CI, 0.99–2.91], p = 0.05) compared with all other patients.ConclusionOur findings suggest that the combination of low sPD-1 and high sPD-L1 levels is a potential negative biomarker for PD-1 blockade therapy.

Introduction: The comprehensive complication index (CCI), which weights all postoperative complications according to severity and integrates them into a single formula, has been reported as a new evaluation system. We aimed to compare the CCI with the Clavien‒Dindo Classification (CDC) to patients with ulcerative colitis (UC). Methods: Patients who underwent initial surgery for UC from April 2012 to March 2020 were included. The patients were classified into a length of stay (LOS) >30 days group or an LOS ≤30 days group. We performed a multivariate analysis of risk factors for LOS >30 days in the model with the factors identified in the univariate analysis plus the CCI (the CCI model) and plus CDC (the CDC model). An ROC curve was used to test the difference in the area under the curve (AUC) between the CCI model and the CDC model. Results: The median LOS was 21 days (IQR: 16-29 days), and the rate of LOS >30 days was 119/588 (20.2%). In the CCI model, age at the time of surgery (OR=1.24, 95% CI 1.07-1.45, p=0.01), ASA score ≥3 (OR=1.94, 95% CI 1.00-3.76, p=0.04), and CCI (OR=1.07, 95% CI 1.05-1.09; p30 days. The AUC value of the CCI model (0.86) was significantly better in relation to LOS > 30 days than that of the CDC model (0.82) (p =0.02). Conclusion: The CCI was a better measure of LOS than was the CDC and was found to be a useful indicator in UC.

Abstract Background Clinicians, researchers, and patients alike would greatly benefit from more accessible and inexpensive biomarkers for neural β-amyloid (Aβ). We aimed to assess the performance of fully automated plasma Aβ immunoassays, which correlate significantly with immunoprecipitation mass spectrometry assays, in predicting brain Aβ status as determined by visual read assessment of amyloid positron emission tomography (PET). Methods The plasma Aβ42/Aβ40 ratio was measured using a fully automated immunoassay platform (HISCL series) in two clinical studies (discovery and validation studies). The discovery and validation sample sets were retrospectively and randomly selected from participants with early Alzheimer’s disease (AD) identified during screening for the elenbecestat Phase 3 program. Results We included 197 participants in the discovery study (mean [SD] age 71.1 [8.5] years; 112 females) and 200 in the validation study (age 70.8 [7.9] years; 99 females). The plasma Aβ42/Aβ40 ratio predicted amyloid PET visual read status with areas under the receiver operating characteristic curves of 0.941 (95% confidence interval [CI] 0.910–0.973) and 0.868 (95% CI 0.816–0.920) in the discovery and validation studies, respectively. In the discovery study, a cutoff value of 0.102 was determined based on maximizing the Youden Index, and the sensitivity and specificity were calculated to be 96.0% (95% CI 90.1–98.9%) and 83.5% (95% CI 74.6–90.3%), respectively. Using the same cutoff value, the sensitivity and specificity in the validation study were calculated to be 88.0% (95% CI 80.0–93.6%) and 72.0% (95% CI 62.1–80.5%), respectively. Conclusions The plasma Aβ42/Aβ40 ratio measured using the HISCL series achieved high accuracy in predicting amyloid PET status. Since our blood-based immunoassay system is less invasive and more accessible than amyloid PET and cerebrospinal fluid testing, it may contribute to the diagnosis of AD in routine clinical practice.

Abstract The streptavidin–biotin system is known to have a very high affinity and specificity and is widely used in biochemical immunoassays and diagnostics. However, this method is affected by endogenous D-biotin in serum sample measurements (biotin interference). While several efforts using alternative high-affinity binding systems (e.g., genetically modified streptavidin and biotin derivatives) have been attempted, these efforts have all led to reduction in affinity. To solve this interference issue, the enantiomer of streptavidin was synthesized, which enabled specific binding to L-biotin. We successfully obtained a functional streptavidin molecule by peptide synthesis using D-amino acids and an in vitro folding technique. Several characterizations, including size exclusion chromatography (SEC), circular dichroism spectra (CD), and heat denaturation experiments collectively confirmed the higher-order enantiomer of natural streptavidin had been formed with comparable stability to the natural protein. L-biotin specific binding of this novel molecule enabled us to avoid biotin interference in affinity measurements using the Biacore system and enzyme-linked immunosorbent assay (ELISA). We propose the enantiomer of streptavidin as a potential candidate to replace the natural streptavidin–biotin system, even for in vivo use.

High temperatures lead to DNA double-strand break accumulation and asynapsis that trigger the meiotic checkpoint and apoptosis of spermatocytes in ex vivo cultures, providing a rationale for the heat sensitivity of sperm production in mammals.

Abstract The naticid snail Laguncula pulchella is an invasive species that preys on clams in tidal flats and has serious impacts on clam fisheries in Japan. Laguncula pulchella burrow in sand, but often crawl on sediment surfaces during low tide. We investigated seasonal changes in the abundance and sex ratio of crawling L. pulchella during the daytime at Matsukawaura Lagoon, Japan, from March to October from 2015 to 2019. The density of crawling individuals peaked in July. The sex ratio of crawling individuals varied with months and years but was significantly biased towards males during the main copulation period (July–August); males accounted for 77–98% of the mature crawling individuals (≥ 25 mm shell height). The somatic condition of mature males declined from June to August, whereas that of females was constant during this period. These results indicate that mature males actively come to the sand surface during low tide to search for females for copulation from July to August. Fishermen make efforts to remove crawling individuals in summer, but the male-biased sex ratio must also be considered for effective population control of this species.

Abstract While mRNA vaccines against SARS-CoV-2 are exceedingly effective in preventing symptomatic infection, their immune response features remain to be clarified. In the present prospective study, 225 healthy individuals in Japan, who received two BNT162b2 doses, were enrolled. Correlates of BNT162b2-elicited SARS-CoV-2-neutralizing activity (50% neutralization titer: NT50; assessed using infectious virions) with various determinants were examined and the potency of sera against variants of concerns was determined. Significant rise in NT50s was seen in sera on day 28 post-1st dose. A moderate inverse correlation was seen between NT50s and ages, but no correlation seen between NT50s and adverse effects. NT50s and SARS-CoV-2-S1-binding-IgG levels on day 28 post-1st dose and pain scores following the 2nd dose were greater in women than in men. The average half-life of NT50s was ~ 68 days, and 23.6% (49 out of 208 individuals) failed to show detectable neutralizing activity on day 150. While sera from elite-responders (NT50s > 1,500: the top 4% among the participants) potently to moderately blocked all variants of concerns examined, some sera with low NT50s failed to block the B.1.351-beta strain. Since BNT162b2-elicited immunity against SARS-CoV-2 is short, an additional vaccine or other protective measures are needed.

Although nuclear power plant accidents have raised concerns about the genetic consequences of radiation exposure, no studies have successfully compared the DNA of wild animals before and after the accident in the same area around a nuclear power plant. Here, we investigated 12 microsatellites of Japanese wild boar in the Fukushima prefecture before and after the Fukushima Daiichi nuclear accident. A population structure analysis of the microsatellites revealed that wild boar of the 4 prefectures of North Kanto were divided into 5 populations before the nuclear accident. Yet, wild boar sampled after the nuclear accident did not belong to any of these 5 populations. The reason for this result was related to significant changes in the composition of microsatellites after the nuclear accident; on average, 90% of post-accident microsatellite alleles were newly emergent in Fukushima prefecture. In samples obtained before the nuclear accident, each microsatellite had either even or odd-length alleles. Yet, in samples obtained after the nuclear accident, most microsatellites had alleles of both even and odd lengths. Moreover, we examined haplotypes and their frequencies in the mitochondrial DNA control region, and found no significant differences before and after the nuclear accident in both Fukushima and Tochigi prefectures, and no haplotypes were distributed only in distant areas suggesting an influx of wild boar after the nuclear accident. This research suggested that the change in microsatellites was a result of microsatellite instability rather than a result of the migration of wild boar from other areas. This research provides important insights into the effects of a nuclear accident on mammals.

Homing peptides to the spinal cord were identified and isolated using phage display technology. In vivo biopanning was performed by intravenous systemic injection of a phage library to screen specific peptides targeting the spinal cord of mice. Analyses of the sequences of targeted phages yielded two candidate peptides targeting the spinal cord: SP1 (C-LHQSPHI-C) and SP2 (C-PTNNPRS-C). These peptides were synthesized and intravenously injected into mice to evaluate their tissue specificity and potential as gene delivery carriers. The complexes between SP1 or SP2 peptides and the plasmid vector expressing the reporter gene could induce gene transduction in the spinal cord through systemic injection without gene expression in the brain, liver, and kidney. In addition, intravenous injection of the complex between SP1 and the vectors induced interleukin-4 expression in the spinal cord, resulting in effective suppression of lipopolysaccharide-induced hyperalgesia. Therefore, intravenously administered spinal cord homing peptides complexed with a plasmid vector provided tissue-specific treatment featuring gene delivery to the CNS through systemic circulation. This novel method of gene delivery is feasible and has great potential for clinical application. Keywords: homing peptides, gene delivery, spinal cord, systemic circulation, targeting

A disintegrin and metalloprotease 17 (ADAM17) catalyzes the cleavage and release of the ectodomains of its substrates at the cell surface in a process termed ectodomain shedding. However, not all ADAM17 substrates have been identified. Here, we used cell surface protein-specific labeling and proteomic approaches to detect and identify ADAM17 substrates. HeLa cell surface proteins were labeled with a fluorescent dye and cultured with or without TAPI-2, an ADAM17 inhibitor. Labeled proteins released into the culture medium were detected by 2-dimensional gel electrophoresis (2DE). Protein spots showing decreased intensity in response to TAPI-2 were selected as substrates of ADAM17 or their binding proteins, and identified by mass spectrometry. ADAM17 knockdown was preformed to examine the behavior of identified proteins. Of 347 proteins detected by 2DE, 49 showed lower intensity in TAPI-2 (+) than in TAPI-2 (-) samples (p < 0.05), and were considered as candidate substrates of ADAM17. Mass spectrometric analysis of 14 protein spots showing >50% decreased intensity identified clusterin as a novel ADAM17 substrate, in addition to known substrates such as desmoglein-2. Western blot analysis showed that ADAM17 knockdown decreased the levels of clusterin fragments cleaved and released from the cell surface. The results identified clusterin as a novel ADAM17 substrate. The method used to identify clusterin could be used to identify the substrates of other sheddases involved in ectodomain shedding.

The efficiency of in vitro platelet production is considerably low compared with physiological activity due to the lack of pivotal factors that are essential in vivo. We developed an ex vivo platelet production system, introducing human megakaryocytes into an isolated porcine thighbone and culturing in closed circuit. The efficiency of the ex vivo platelet production system was compared to those in vivo and in vitro. CD61+ platelet-like cells were counted by immunostaining and flow cytometry. Results showed that 4.41 ± 0.27 × 103 CD61+ platelet-like cells were produced by 1 × 103 megakaryocytes in the ex vivo system, while 3.80 ± 0.87 × 103 and 0.12 ± 0.02 × 103 were produced in the in vivo and in vitro systems, respectively. Notably, ex vivo and in vitro production systems generated cells that responded well to thrombin stimulation and expressed functional molecules, such as CD62P. Overall, our ex vivo production system was comparable to in vivo production system and produced platelet-like cells that were functionally superior to those produced in vitro. In future, the present ex vivo production system implementing xenogeneic bone marrow would offer a promising alternative for industrial-scale production of platelet-like cells.

Abstract Background To understand the roles of serum exosomes in rheumatoid arthritis (RA), we comprehensively investigated the protein profiles of serum exosomes in patients with RA. Methods Exosomes were isolated from serum samples obtained from 33 patients (12 with active RA [aRA], 11 with inactive RA [iRA], 10 with osteoarthritis [OA]) and 10 healthy donors (HLs). Proteins extracted from the exosomes were separated by two-dimensional differential gel electrophoresis (2D-DIGE) and identified by mass spectrometry. Results In total, 204 protein spots were detected by 2D-DIGE. In the aRA, iRA, and OA groups, 24, 5, and 7 spots showed approximately ≥ ±1.3-fold intensity differences compared with the HL group, respectively. We were able to identify proteins in six protein spots. Among them, the protein spot identified as Toll-like receptor 3 (TLR3) showed approximately 6-fold higher intensity in the aRA group than in the other groups. Conclusions Patients with active RA possessed considerably different protein profiles of serum exosomes from patients with iRA, patients with OA, and healthy donors. The unique protein profile of serum exosomes, such as the possession of abundant TLR3 fragments, may reflect the pathophysiology of active RA.

Astrocyte- and microglia-targeting peptides were identified and isolated using phage display technology. A series of procedures, including three cycles of both in vivo and in vitro biopanning, was performed separately in astrocytes and in M1 or M2 microglia, yielding 50–58 phage plaques in each cell type. Analyses of the sequences of this collection identified one candidate homing peptide targeting astrocytes (AS1[C-LNSSQPS-C]) and two candidate homing peptides targeting microglia (MG1[C-HHSSSAR-C] and MG2[C-NTGSPYE-C]). To determine peptide specificity for the target cell in vitro, each peptide was synthesized and introduced into the primary cultures of astrocytes or microglia. Those peptides could bind to the target cells and be selectively taken up by the corresponding cell, namely, astrocytes, M1 microglia, or M2 microglia. To confirm cell-specific gene delivery to M1 microglia, the complexes between peptide MG1 and siRNA-interferon regulatory factor 5 were prepared and intrathecally injected into a mouse model of neuropathic pain. The complexes successfully suppressed hyperalgesia with high efficiency in this neuropathic pain model. Here, we describe a novel gene therapy for the treatment neuropathic pain, which has a high potential to be of clinical relevance. This strategy will ensure the targeted delivery of therapeutic genes while minimizing side effects to non-target tissues or cells.

BackgroundContinuous glucose monitoring (CGM) is reported to be a useful technique, but difficult or inconvenient for some patients and institutions. We are developing a glucose area under the curve (AUC) monitoring system without blood sampling using a minimally invasive interstitial fluid extraction technology (MIET). Here we evaluated the accuracy of interstitial fluid glucose (IG) AUC measured by MIET in patients with diabetes for an extended time interval and the potency of detecting hyperglycemia using CGM data as a reference.MethodsThirty-eight inpatients with diabetes undergoing CGM were enrolled. MIET comprised a pretreatment step using a plastic microneedle array and glucose accumulation step with a hydrogel patch, which was placed on two sites from 9:00 AM to 5:00 PM or from 10:00 PM to 6:00 AM. IG AUC was calculated by accumulated glucose extracted by hydrogel patches using sodium ion as standard. ResultsA significant correlation was observed between the predicted AUC by MIET and CGM in daytime (r=0.76) and nighttime (r=0.82). The optimal cutoff for the IG AUC value of MIET to predict hyperglycemia over 200 mg/dL measured by CGM for 8 hours was 1,067.3 mg·hr/dL with 88.2% sensitivity and 81.5% specificity.ConclusionWe showed that 8-hour IG AUC levels using MIET were valuable in estimating the blood glucose AUC without blood sampling. The results also supported the concept of using this technique for evaluating glucose excursion and for screening hyperglycemia during 8 hours in patients with diabetes at any time of day.

Background/Aims: Recent genome-wide analyses have provided strong evidence concerning adverse events caused by thiopurine drugs such as azathioprine (AZA) and 6-mercaptopurine. The strong associations identified between NUDT15 p.Arg139Cys and thiopurine-induced leukopenia and severe hair loss have been studied and confirmed over the last 2 years. However, other coding variants, including NUDT15 p.Val18_Val19insGlyVal, NUDT15 p.Val18Ile, and FTO p.Ala134Thr, and a noncoding variation in RUNX1 (rs2834826) remain to be examined in detail in this respect. Therefore, we investigated the correlation between these adverse events and the 5 recently identified variants mentioned above among Japanese patients with inflammatory bowel diseases (IBD).Methods: One hundred sixty thiopurine-treated patients with IBD were enrolled. Genotyping was performed using TaqMan SNP Genotyping Assays or Sanger sequencing.Results: None of the 5 variants were associated with gastrointestinal intolerance to AZA. However, NUDT15 p.Arg139Cys was significantly associated with the interval between initiation and discontinuation of AZA among patients with gastrointestinal intolerance. This variant was strongly associated with early (

BackgroundManagement of postprandial hyperglycemia is a key aspect in diabetes treatment. We developed a novel system to measure glucose area under the curve (AUC) using minimally invasive interstitial fluid extraction technology (MIET) for simple monitoring of postprandial glucose excursions. In this study, we evaluated the relationship between our system and continuous glucose monitoring (CGM) by comparing glucose AUC obtained using MIET with that obtained using CGM for a long duration.MethodsTwenty diabetic inpatients wearing a CGM system were enrolled. For MIET measurement, a plastic microneedle array was applied to the skin as pretreatment, and hydrogels were placed on the pretreated area to collect interstitial fluid. Hydrogels were replaced every 2 or 4 hours and AUC was predicted on the basis of glucose and sodium ion levels.ResultsAUC predicted by MIET correlated well with that measured by CGM (r=0.93). Good performances of both consecutive 2- and 4-hour measurements were observed (measurement error: 11.7%±10.2% for 2 hours and 11.1%±7.9% for 4 hours), indicating the possibility of repetitive measurements up to 8 hours. The influence of neither glucose fluctuation nor average glucose level over the measurement accuracy was observed through 8 hours.ConclusionOur system showed good relationship with AUC values from CGM up to 8 hours, indicating that single pretreatment can cover a large portion of glucose excursion in a day. These results indicated possibility of our system to contribute to convenient monitoring of glucose excursions for a long duration.

We developed a minimally invasive glucose monitoring system that uses a microneedle to permeate the skin surface and a small hydrogel to accumulate interstitial fluid glucose. The measurement of glucose and sodium ion levels in the hydrogel is required for estimating glucose levels in blood; therefore, we developed a small, enzyme-fixed glucose sensor with a high-selectivity, all-solid-state, sodium ion-selective electrode (ISE) integrated into its design. The glucose sensor immobilized glucose oxidase showed a good correlation between the glucose levels in the hydrogels and the reference glucose levels (r > 0.99), and exhibited a good precision (coefficient of variation = 2.9%, 0.6 mg/dL). In the design of the sodium ISEs, we used the insertion material Na0.33MnO2 as the inner contact layer and DD16C5 exhibiting high Na+/K+ selectivity as the ionophore. The developed sodium ISE exhibited high selectivity (\( \log \,k^{pot}_{Na,K} = -2.8\)) and good potential stability. The sodium ISE could measure 0.4 mM (10−3.4 M) sodium ion levels in the hydrogels containing 268 mM (10−0.57 M) KCl. The small integrated sensor (ϕ < 10 mm) detected glucose and sodium ions in hydrogels simultaneously within 1 min, and it exhibited sufficient performance for use as a minimally invasive glucose monitoring system.

In this study, we investigated the geographical distribution of haplotypes of Cardiocondyla kagutsuchi Terayama in Japan using COI/II mitochondrial DNA. We also examined their genealogy with C. kagutsuchi in other areas and their close relative species. Four haplotypes were found. While two of them were found in a limited area (Ishigaki and Okinawa Islands) separately, the others were distributed widely across Honsyu, Shikoku, and Kyusyu areas in Japan. The newly invaded area by C. kagutsuchi in Japan was Shizuoka prefecture. Their haplotype of Shizuoka were the same as the two haplotypes of the Honsyu, Shikoku, and Kyusyu areas. The haplotype network showed that the two haplotypes were distant from each other. The distance between them was 33, even though the two haplotypes are distributed in the same area. From the phylogenetic tree that we constructed, we found that C. strigifrons was in the same clade as C. kagutsuchi.

In recent years, attention has been increasingly devoted to the development of rescue robots that can protect humans from the inherent risks of rescue work. Particularly, anticipated is the development of a robot that can move deeply through small spaces. We have devoted our attention to peristalsis, the movement mechanism used by earthworms. A reinforcement learning technique used for the derivation of the robot movement pattern, Q-learning, was used to develop a three-segmented peristaltic crawling robot with a motor drive. Characteristically, peristalsis can provide movement capability if at least three segments work, even if a segmented part does not function. Therefore, we had intended to derive the movement pattern of many-segmented peristaltic crawling robots using Q-learning. However, because of the necessary increase in calculations, in the case of many segments, Q-learning cannot be used because of insufficient memory. Therefore, we devoted our attention to a learning method called Actor–Critic, which can be implemented with low memory. Because Actor-Critic methods are TD methods that have a separate memory structure to explicitly represent the policy independent of the value function. Using it, we examined the movement patterns of six-segmented peristaltic crawling robots.

Sialic acid-binding lectin from Rana catesbeiana eggs (cSBL) is a multifunctional protein that has lectin and ribonuclease activity. In this study, the anti-tumor activities of cSBL were assessed using a panel of breast cancer cell lines. cSBL suppressed the cell growth of all cancer cell lines tested here at a concentration that is less toxic, or not toxic at all, to normal cells. The growth suppressive effect was attributed to the cancer-selective induction of apoptosis. We assessed the expressions of several key molecules associated with the breast cancer phenotype after cSBL treatment by western blotting. cSBL decreased the expression level of estrogen receptor (ER) α, while it increased the phosphorylation level of p38 mitogen-activated protein kinase (MAPK). cSBL also suppressed the expression of the progesterone receptor (PgR) and human epidermal growth factor receptor type 2 (HER2). Furthermore, it was revealed that cSBL decreases the expression of the epidermal growth factor receptor (EGFR/HER1) in triple-negative breast cancer cells. These results indicate that cSBL induces apoptosis with decreasing ErbB family proteins and may have great potential for breast cancer chemotherapy, particularly in triple-negative phenotype cells.

Passive dynamic locomotion occurs through the interaction of an object’s dynamics and the ground, using no actuators, sensors, or controls. Passive dynamics is important for exploring the principles of human and animal locomotion. An important characteristic is that there might be a stable limit cycle for passive dynamic locomotion. Expression of stable limit cycles in passive bipedal walking, bipedal running, and quadrupedal running has been confirmed. However, studies conducted to date have characterized passive monopedal hopping as unstable. We investigated the dynamic stability of the passive monopedal hopping without torso pitch. Numerical simulation revealed that the passive monopedal hopping examined in this study has a stable limit cycle. Moreover, both theoretical analysis and simulation results suggest that asymptotically stable fixed points of the 1-PDH model are related closely to the leg spring damping ratio and the hopping period. Results show that the 1-PDH model has high stability with respect to steps. Additionally, findings indicate that changes in leg spring stiffness cause changes in forward velocity during stable hopping with no feedback control.

In this paper, we propose a double air chambers artificial muscle to eliminate the hysteresis in the extension and contraction movement of pneumatic artificial muscles. In this paper, the basic structure of the double air chambers artificial muscle is a rubberless artificial muscle with a particularly large hysteresis loop. The double air chambers artificial muscle aims to eliminate hysteresis by directly pressurizing the inside and outside of the air chamber and actively deforming the air chamber. The hysteresis is reduced by the pressure outside of the air chamber (external pressure). Since the appropriate external pressure varies depending on the contraction force and amount of contraction, we proposed a method to regulate the appropriate external pressure by feedback control. The experimental results show that hysteresis was eliminated in the static characteristics. It was also found that the output gain decreased, and the phase lag increased as the target frequency increased. The output gain did not change with increasing load. The phase lag tended to improve with the PID controller compared to the PI controller. These results suggest that the combination of double air chambers artificial muscle and external pressure-regulated feedback control can achieve non-hysteresis position control, and it is useful as an actuator in mechatronic systems.

Ants are known to show diverse mating behaviors across different species, but detailed reports on such behaviors are limited to only 0.7% of ant species. Therefore, we report on the mating behavior of the alate females and males in the small carpenter ant, Camponotus yamaokai. Its process was classified into two phases: mating attempt, and copulation. Out of 53 pairs we observed, 23 pairs engaged in a total of 36 mating attempts. The mating attempt durations in cases where copulation failed were significantly longer than those leading to copulation. Among these attempts, copulation was observed 10 times in 9 pairs, including multiple mating in one pair. The copulation lasted for 24.0 ± 5.1 seconds (mean ± SD). At the end of copulation, males pulled out their genitalia quickly to avoid potential attacks from the females. This behavior might be a strategy of males to minimize the risk of mortality during a single mating and support them to engage in multiple mating.

Aims: This study aimed to clarify the clinical characteristics of Pneumocystis jirovecii pneumonia (PJP) infection in patients with ulcerative colitis (UC) and to identify risk factors for PJP using a retrospective case-control study. Methods: Of 4,525 patients with UC treated between 2007 and 2019, we identified those who satisfied the criteria for PJP. The Lichtiger clinical activity index (LCI) was compared between the initiation of immunosuppressive drug treatment and the onset of PJP. A retrospective case-control study was conducted using a PJP group and a non-PJP group. Results: Nine patients experienced PJP, of whom two died. Since October 2014, there were no cases of PJP among UC patients aged =50 years who were prescribed three or more immunosuppressive agents given prophylactic sulfamethoxazole-trimethoprim (TPM-SMX). The median LCI (range) was 13 (8-17) at the initiation of treatment versus 2 (1-8) at PJP onset (p = 0.016). The median time to PJP onset was 83 days after treatment initiation. In the PJP group the median age was significantly greater (p = 0.022), three immunosuppressants were used significantly more frequently (p = 0.004), and the lymphocyte counts during treatment were significantly lower (p < 0.01) than in the non-PJP group. The cut-off lymphocyte count that distinguished PJP patients from non-PJP patients was 570/µL according to a receiver-operating curve analysis. Conclusions: Prophylactic administration of TPM-SMX prevented further cases of PJP. The onset of PJP occurred at the same time as the symptoms of UC were stabilizing and the immunosuppressive drugs were being reduced. Greater age, lower lymphocyte count, and treatment with three immunosuppressive drugs were risk factors for PJP. [ABSTRACT FROM AUTHOR]

Understanding concentration distributions, release sites, and release dynamics of volatile organic compounds (VOCs) from the human is expected to lead to methods for noninvasive disease screening and assessment of metabolisms. In this study, we developed a visualization system (sniff-cam) that enabled one to identify a spatiotemporal change of gaseous acetaldehyde (AcH) in real-time. AcH sniff-cam was composed of a camera, a UV-LED array sheet, and an alcohol dehydrogenase (ADH)-immobilized mesh. A reverse reaction of ADH was employed for detection of gaseous AcH where a relationship between fluorescence intensity from nicotinamide adenine dinucleotide and the concentration of AcH was inversely proportional; thus, the concentration distribution of AcH was measured by detecting the fluorescence decrease. Moreover, the image differentiation method that calculated a fluorescence change rate was employed to visualize a real-time change in the concentration distribution of AcH. The dynamic range of the sniff-cam was 0.1-10 ppm which encompassed breath AcH concentrations after drinking. Finally, the sniff-cam achieved the visualization of the concentration distribution of AcH in breath and skin gas. A clear difference of breath AcH concentration was observed between aldehyde dehydrogenase type 2 active and inactive subjects, which was attributed to metabolic capacities of AcH. AcH in skin gas showed a similar time course of AcH concentration to the breath and a variety ofrelease concentration distribution. Using different NADH-dependent dehydrogenases in the sniff-cam could lead to a versatile method for noninvasive disease screening by acquiring spatiotemporal information on various VOCs in breath or skin gas. [ABSTRACT FROM AUTHOR]

Various volatile organic compounds can be found in human transpiration, breath and body odor. In this paper, a novel two-dimensional fluorometric imaging system, known as a "sniffer-cam" for ethanol vapor released from human breath and palm skin was constructed and validated. This imaging system measures ethanol vapor concentrations as intensities of fluorescence through an enzymatic reaction induced by alcohol dehydrogenase (ADH). The imaging system consisted of multiple ultraviolet light emitting diode (UV-LED) excitation sheet, an ADH enzyme immobilized mesh substrate and a high-sensitive CCD camera. This imaging system uses ADH for recognition of ethanol vapor. It measures ethanol vapor by measuring fluorescence of nicotinamide adenine dinucleotide (NADH), which is produced by an enzymatic reaction on the mesh. This NADH fluorometric imaging system achieved the two-dimensional real-time imaging of ethanol vapor distribution (0.5-200 ppm). The system showed rapid and accurate responses and a visible measurement, which could lead to an analysis of metabolism function at real time in the near future. [ABSTRACT FROM AUTHOR]

This paper describes a device to support a patient's motion of their upper limbs. The device has three functions in a single system: power assistance, rehabilitation, and meal support. The former two functions have been described in a previous study. In this paper, designs and evaluations of the meal supporting function considering maintaining residual muscular strength are presented. The mechanical design is based on human body statistical data. The controller consists of an electromyogram signal-based interface used for maintaining muscular strength. We evaluate the effectiveness of the meal supporting mode through some experimentation.

Shortage of labor and increased work of young people are causing problems in terms of care and welfare of a growing proportion of elderly people. This is a looming social problem because people of advanced ages are increasing. Necessary in the fields of care and welfare, pneumatic artificial muscles in actuators of robots are being examined. Pneumatic artificial muscles have a high output per unit of weight, and they are soft, similarly to human muscles. However, in previous research of robots using pneumatic artificial muscles, rigid sensors were often installed at joints and other locations due to the robots’ structures. Therefore, we developed a smart actuator that integrates a bending sensor that functions as a human muscle spindle; it can be externally attached to the pneumatic artificial muscle. This paper reports a smart artificial muscle actuator that can sense contraction, which can be applied to developed self-monitoring and robot posture control.

The genus Vigna includes legume crops such as cowpea, mungbean and azuki bean, as well as >100 wild species. A number of the wild species are highly tolerant to severe environmental conditions including high-salinity, acid or alkaline soil; drought; flooding; and pests and diseases. These features of the genus Vigna make it a good target for investigation of genetic diversity in adaptation to stressful environments; however, a lack of genomic information has hindered such research in this genus. Here, we present a genome database of the genus Vigna, Vigna Genome Server ('VigGS', http://viggs.dna.affrc.go.jp), based on the recently sequenced azuki bean genome, which incorporates annotated exon-intron structures, along with evidence for transcripts and proteins, visualized in GBrowse. VigGS also facilitates user construction of multiple alignments between azuki bean genes and those of six related dicot species. In addition, the database displays sequence polymorphisms between azuki bean and its wild relatives and enables users to design primer sequences targeting any variant site. VigGS offers a simple keyword search in addition to sequence similarity searches using BLAST and BLAT. To incorporate up to date genomic information, VigGS automatically receives newly deposited mRNA sequences of pre-set species from the public database once a week. Users can refer to not only gene structures mapped on the azuki bean genome on GBrowse but also relevant literature of the genes. VigGS will contribute to genomic research into plant biotic and abiotic stresses and to the future development of new stress-tolerant crops. [ABSTRACT FROM AUTHOR]

Thermal transport across organic-inorganic interfaces is fundamental to understanding heat transfer in polymer-based composites, microelectronics, and energy conversion systems. We used the Langmuir-Blodgett (LB) technique to deposit nanometer-thick films of poly(vinyl acetate) (PVAc) on silicon and gold substrates in two distinct states: Liquid condensed (Lc) and Liquid expanded (Le). We used frequency domain thermoreflectance to measure the thermal conductivity of the PVAc film and its thermal interfaceconductance to the substrate. We found that PVAc films prepared through the LB process have a higher thermal conductivity when compared to bulk. We measured the thermal interfaceconductance between PVAc and gold to be approximately 90 MW/m² K for both the Le and Lc states, and the thermal interfaceconductance between PVAc and silicon to be approximately 70 MW/m² K for both the Le and Lc states. [ABSTRACT FROM AUTHOR]

We developed a minimally invasive glucose monitoring system that uses a microneedle to permeate the skin surface and a small hydrogel to accumulate interstitial fluid glucose. The measurement of glucose and sodium ion levels in the hydrogel is required for estimating glucose levels in blood; therefore, we developed a small, enzyme-fixed glucose sensor with a high-selectivity, all-solid-state, sodium ion-selective electrode (ISE) integrated into its design. The glucose sensor immobilized glucose oxidase showed a good correlation between the glucose levels in the hydrogels and the reference glucose levels (r > 0.99), and exhibited a good precision (coefficient of variation = 2.9%, 0.6 mg/dL). In the design of the sodium ISEs, we used the insertion material Na0.33MnO2 as the inner contact layer and DD16C5 exhibiting high Na+ /K+ selectivity as the ionophore. The developed sodium ISE exhibited high selectivity (logkpot NNa,K = -2.8) and good potential stability. The sodium ISE could measure 0.4 mM (10-3.4 M) sodium ion levels in the hydrogels containing 268 mM (10 -0.57(M) KCl. The small integrated sensor (Φ < 10 mm) detected glucose and sodium ions in hydrogels simultaneously within 1 min, and it exhibited sufficient performance for use as a minimally invasive glucose monitoring system. [ABSTRACT FROM AUTHOR]

Objective: To find a blood biomarker and disease-related peptides in Alzheimer’s disease (AD), we comprehensively detected serum peptides. Methods: Ion intensity of serum peptides from 62 AD patients and 82 control subjects was measured by mass spectrometry. Results: A total of 157 peptides were detected from 30 AD patients and 30 healthy control (HC) subjects. Sixty out of the 157 peptide profiles discriminated between the AD and HC groups. Sixteen out of the 60 peptides were identified, 10 out of which were fragments of a fibrinogen α chain (FIBA). Among the 10 peptides, four and six peptides were derived from fibrinopeptide A (FPA, A α1--16) and the C-terminal region of the αC-domain (αCDC, A α557-610), respectively. The profile of 10 FIBA-derived peptides combined with age discriminated between the two groups with an area under the receiver operating characteristic curve (AUROC) of 0.940. Validation of this model using a testing set of 32 AD patients and 19 HC subjects showed an AUROC of 0.717, sensitivity of 65.6%, and specificity of 73.7% by a cutoff value of 0.56420. Another value, 0.04029, showed sensitivity of 96.9%, suggesting that subjects with values less than 0.04029 rarely possess AD. FPA and αCDC showed increased ion intensity in the AD group compared with the HC group (p<0.05). Conclusions: The profile of 10 FIBA-derived peptides combined with age would be a candidate biomarker for AD, which facilitates screening of the disease. The significant release of FPA and αCDC may be involved in the aberrant coagulation that leads to vascular damage in AD. [ABSTRACT FROM AUTHOR]

Background: Protein-protein interaction (PPI) plays a core role in cellular functions. Massively parallel supercomputing systems have been actively developed over the past few years, which enable large-scale biological problems to be solved, such as PPI network prediction based on tertiary structures. Results: We have developed a high throughput and ultra-fast PPI prediction system based on rigid docking, "MEGADOCK", by employing a hybrid parallelization (MPI/OpenMP) technique assuming usages on massively parallel supercomputing systems. MEGADOCK displays significantly faster processing speed in the rigid-body docking process that leads to full utilization of protein tertiary structural data for large-scale and network-level problems in systems biology. Moreover, the system was scalable as shown by measurements carried out on two supercomputing environments. We then conducted prediction of biological PPI networks using the post-docking analysis. Conclusions: We present a new protein-protein docking engine aimed at exhaustive docking of mega-order numbers of protein pairs. The system was shown to be scalable by running on thousands of nodes. The software package is available at: http://www.bi.cs.titech.ac.jp/megadock/k/. [ABSTRACT FROM AUTHOR]

Continuous hydrothermal synthesis of Fe2O3nanoparticles was carried out at 673 K and 30 MPa. Two types of mixers, a conventional T-type micromixer and a central collision-type micromixer (CCM), were used for the synthesis. CCM was newly fabricated on the basis of the concept for preventing heterogeneous nucleation induced on the inner wall of the mixer. Residence time and Fe(NO3)3molality of the starting solution varied from 0.1 to 1.0 s and 0.05 to 0.50 mol/kg, respectively. Effects of the mixer structure, the residence time, and the Fe(NO3)3molality on crystal structure, Fe3犉묢, average particle size, and size distribution were discussed. CCM effectively worked for producing smaller Fe2O3nanoparticles with narrow and monomodal distribution. [ABSTRACT FROM AUTHOR]

Ferrite nanoparticles from Fe(NO3)3and Me(NO3)2(Me Ni, Cu, Zn) aqueous solutions could be synthesized continuously with a flow reactor at 673 K, 30 MPa, and 4 s residence time. The particles were characterized by TEM, XRD, and ICP to obtain sizes, crystal structures, lattice parameters, and Me/Fe molar ratios. Solid-solution nanoparticles of MeFe2O4and -Fe2O3with a cubic spinel structure and an average particle size under 10 nm were obtained. Conversion of Fe3was more than 0.97 at the given residence time, and conversions of Me2increased from 0 to 0.62 with increasing residence time. The particle size increased with increasing residence time, and no significant difference in divalent cations was observed. The Me/Fe molar ratio in the obtained solid-solution nanoparticles of MeFe2O4and -Fe2O3increased with increasing residence time and KOH molality. The Me/Fe molar ratio also increased in the order Zn < Cu < Ni at a given residence time, and this trend could be explained on the basis of ZnO, CuO, and NiO solubilities. The increase of the size and the Me/Fe molar ratio in the obtained particles with increasing residence time shows that, in the formation mechanism, primary solid-solution particles with low Me/Fe molar ratios probably nucleated and, after surface dissolution of the particles, dissolved Fe3recrystallized on the surface with incorporation of Me2. [ABSTRACT FROM AUTHOR]

Abstract  We herein report a case of gastric small cell carcinoma (GSCC), which was successfully treated by surgery and postoperative chemotherapy consisting of cisplatin (CDDP) and the fluoropyrimidine S-1. The patient was a 63-year-old man in whom a gastric tumor had been endoscopically detected. The type 1 tumor was located in the gastric body. An abdominal computed tomogram showed many metastasized nodes around the stomach. A total gastrectomy with regional node dissection was performed. The removed tumor was histologically and histochemically diagnosed to be a GSCC with node metastases. Furthermore, washing peritoneal cytology histologically revealed the presence of carcinoma cells. Metastasis was histologically observed in 17 of 24 dissected nodes. After surgery, CDDP was intravenously administered and S-1 was orally administered for 1 year. Consequently, the patient is now well without any recurrence 45 months after surgery. We reviewed 52 Japanese patients with GSCC/endocrine cell carcinoma (EC) reported between 2001 and 2005 with reference to chemotherapy. Chemotherapy using S-1 was performed for 11 of the 52 patients. Four of the 11 patients, including the present case, who were treated with S-1 survived for over 2 years after surgery, although the GSCC/EC of the four patients were staged as III or IV. Therefore, chemotherapy consisting of CDDP and S-1 may provide a survival benefit for patients with GSCC/EC. [ABSTRACT FROM AUTHOR]

This article reviews the preparation and characterization of polycrystalline CdTe and CdZnTe films to be used in large-area, high-sensitivity X-ray panel detectors intended for medical diagnostics. The films, deposited by closed-spaced sublimation, are expected to exhibit excellent efficiency at low X-ray doses because of their high sensitivity. The detectors constructed using these films incorporated a novel hybrid technique, in which zinc-doped CdTe was pre-deposited onto a ceramic substrate and then connected to a TFT circuit substrate. The sensor substrate material was specially selected to avoid both incident X-ray attenuation in the substrate and micro-cracks in the film. Zinc doping, which was used to grow the CdTe film, also served to form the heterojunction diode structure that suppressed leakage current. Moreover, the quality of a polycrystalline CdZnTe film deposited on a 9"×9" substrate was characterized, revealing its applicability to large-area X-ray detectors. Further investigation and improvements are in progress. [ABSTRACT FROM AUTHOR]

Malignant mesothelioma is an aggressive cancer that results from exposure to asbestos. The therapeutic options for this type of cancer are limited; therefore, the development of novel therapeutic agents is urgently required. Sialic acid-binding lectin isolated from Rana catesbeiana oocytes (cSBL) is a novel therapeutic candidate for cancer, which exhibits antitumor activity mediated through RNA degradation. In the present study, we evaluated the effect of cSBL in vitro and in vivo. Xenograft-competent H2452 and MSTO human mesothelioma cell lines were treated with cSBL, and the pathway by which cSBL induces apoptosis was analyzed. In vivo studies were performed using nude mice inoculated with one of the two cell lines, and the effects of cSBL and pemetrexed were monitored simultaneously. Furthermore, the pharmacological interactions between the three agents (pemetrexed, cisplatin and cSBL) were statistically assessed. It was demonstrated that cSBL treatments caused morphological and biochemical apoptotic changes in both cell lines. Caspase cascade analysis revealed that an intrinsic pathway mediated cSBL-induced apoptosis. The administration of cSBL significantly inhibited tumor growth in two xenograft models, without any adverse effects. Furthermore, the combination index and dose reduction index values indicated that the cSBL + pemetrexed combination showed the highest synergism, and thus potential for reducing dosage of each drug, compared with the other combinations, including the existing pemetrexed + cisplatin regimen. cSBL exerted prominent antitumor effects on malignant mesothelioma cells in vitro and in vivo, and showed favorable effects when combined with pemetrexed. These results suggest that cSBL has potential as a novel drug for the treatment of malignant mesothelioma.

Ophraella communa (Coleoptera: Chrysomelidae) is an oligophagous herbivorous beetle that feeds on Ambrosia artemisiifolia. It is native to North America, but was accidentally introduced into Japan in 1995 and Europe in 2013. We analyzed partial DNA sequences of the mitochondrial cytochrome oxidase II gene for O. communa collected from 29 locations in the United States, Japan and Italy. Overall, the results of our analyses indicate that the introduced Japanese populations have lower genetic variation than the native populations. The sequences for the Italian specimens did not share haplotypes with Japanese specimens. These results indicate that the introduced Japanese populations originated from a single introduction, and that the Italian and Japanese populations have different origins.

l-Menthyl α-D-glucopyranosyl-(1→4)- α-D-glucopyranoside (l-α-MenG2), an α-maltoside of l-menthol, was synthesized through a three-step enzymatic reaction. We found that l-α-MenG2 possesses the properties of a low-molecular-weight gelator. Aqueous solutions containing l-α-MenG2 at concentrations above 30 g L-1 show a thermally reversible solgel transition. The solgel transition temperature of the aqueous l-α-MenG2 solution increases with l-α-MenG2 concentration: 12 °C at 30 g L-1 and 24 °C at 250 g L-1. d-Menthyl and d-isomenthyl α-maltosides were also synthesized enzymatically, but their aqueous solutions showed no solgel transition. [ABSTRACT FROM AUTHOR]