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Galactose-deficient (agalactosyl) IgG is significantly increased in the serum of patients with rheumatoid arthritis, and autoantibodies against it are used in clinical tests. Subsequent studies also show increased agalactosyl IgG in many chronic inflammatory diseases. In this study, we generated antibody 42B1 recognizing agalactosyl IgG and developed a new method to evaluate chronic inflammatory diseases with it. Using an ELISA with antibody 42B1, we measured serum levels of agalactosyl IgG in 32 patients with inflammatory bowel disease (IBD), 60 patients with chronic liver disease, 60 patients with chronic pancreatitis, and 32 subjects undergoing health checkups who did not have IBD. Serum agalactosyl IgG levels were increased in all patients with chronic inflammations and partially correlated with clinical parameters. Among the subjects undergoing health checkups, some subjects showed a 15 % elevation of serum agalactosyl IgG levels, suggesting possible latent chronic inflammation. Future studies will examine the 42B1 antibody ELISA in various autoimmune diseases. Altogether, the 42B1 antibody for determination of serum agalactosyl IgG levels is a novel diagnostic tool for chronic inflammation., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Masafumi Shiida and Tatsuya Ono are employees of Minaris Medical Co., Ltd., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Cross-modal interactions between sensory modalities may be necessary for recognition of chewing food by the invisible oral cavity to avoid damaging the tongue and/or oral mucosa. The present study used functional near-infrared spectroscopy to investigate whether the food properties hardness and size influence activities in the posterior parietal cortex and visual cortex during chewing performance in healthy individuals. It was found that an increase in food hardness enhanced both posterior parietal cortex and visual cortex activities, while an increase in food size enhanced activities in the same regions. These findings suggested a quantitative relationship of oral food properties with activities in the brain regions responsible for object recognition and visuospatial processing. It is thus concluded that heightened neural activities in the posterior parietal cortex and visual area reflect an increased demand for cross-modal representation of food properties related to chewing., Competing Interests: Declaration of competing interest Takeshi Uchida and Ikuo Kantake are employed by Dental Support Co., Ltd. Neither has any potential conflicts of interest to declare with respect to the present research, authorship, and/or publication of this article. The remaining authors declare that the research was conducted in the absence of any personal commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2025. Published by Elsevier Inc.)

The poor prognosis of pancreatic cancer is often attributed to difficulties of early detection due to a lack of appropriate risk factors. Previously, we demonstrated the presence of Enterococcus faecalis (E. faecalis) in pancreatic juice and tissues obtained from patients with cancers of the duodeno-pancreato-biliary region, suggesting the possible involvement of this bacterial species in chronic and malignant pancreatic diseases. However, it remains unclear if and how E. faecalis can infect pancreatic ductal cells. In this study, we used immortalized normal human pancreatic ductal epithelial cells (iPDECs) and pancreatic ductal cancer cell lines to demonstrate that E. faecalis adheres to and invades pancreatic ductal lineage epithelial cells. Inhibitors of micropinocytosis or clathrin- or caveolae-mediated endocytosis suppressed iPDEC invasion by E. faecalis. Mechanistically, bacterial expression of enterococcal fibronectin-binding protein A (EfbA) was correlated with adhesive potential of E. faecalis strains. Knockout of fibronectin 1, a binding partner of EfbA, in iPDECs resulted in suppressed E. faecalis adhesion and invasion, suggesting the importance of the EfbA-fibronectin axis in infection of pancreatic ductal epithelial lineage cells. Overall, these results suggest that E. faecalis can colonize pancreatic tissue by infecting iPDECs, at least in part, via the expression of the cell adhesion factor EfbA., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Even though the oral cavity is not visible, food chewing can be performed without damaging the tongue, oral mucosa, or other intraoral parts, with cross-modal perception of chewing possibly critical for appropriate recognition of its performance. This study was conducted to clarify the relationship of chewing food cross-modal perception with cortex activities based on examinations of the posterior parietal cortex (PPC) and visual cortex during chewing in comparison with sham chewing without food, imaginary chewing, and rest using functional near-infrared spectroscopy. Additionally, the effects of a deafferent tongue dorsum on PPC/visual cortex activities during chewing performance were examined. The results showed that chewing food increased activity in the PPC/visual cortex as compared with imaginary chewing, sham chewing without food, and rest. Nevertheless, those activities were not significantly different during imaginary chewing or sham chewing without food as compared with rest. Moreover, subjects with a deafferent tongue dorsum showed reduced PPC/visual cortex activities during chewing food performance. These findings suggest that chewing of food involves cross-modal recognition, while an oral somatosensory deficit may modulate such cross-modal activities., Competing Interests: The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Funders provided support in the form of salaries for authors, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2024 Ishii et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Chronic inflammation of the pancreas is considered to be one of the causes of pancreatic cancer. However, the diagnosis of chronic pancreatitis (CP) is very difficult in the pancreas, where biopsies are difficult to perform. The prevalence of CP is estimated to be many times more common than in patients with actual symptomatic CP. In recent years, abnormal cleavage of certain proteins has attracted attention as a biomarker for CP other than pancreatic enzymes. Connective tissue growth factor (CTGF) is one of the growth factors involved in tissue repair and other processes and is increased by stimulation of transforming growth factor-β, suggesting a relationship of CTGF with fibrosis. In this study, we measured the total length of CTGF in blood and N -terminal fragment CTGF in 48 cases of chronic pancreatitis, 64 cases of pancreatic cancer and 45 healthy volunteers (HV). Interestingly, we found that blood N -terminal fragment CTGF level was significantly increased in CP and pancreatic cancer patients. Multiple logistic regression analysis showed serum levels of N -terminal fragment CTGF, CRP and amylase were significant and independent variables for the differential diagnosis of CP from HV. Receiver operating characteristic analysis showed that area under the curve (AUC) value of serum N -terminal fragment CTGF level was 0.933, which can differentiate between CP and HV. Several factors would be involved in the increase in serum N -terminal fragment CTGF level. In conclusion, serum N -terminal fragment CTGF level is a promising new biomarker for CP., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this article., (© 2024 The Authors.)

Aims: We aimed to investigate the function of an unidentified gene annotated as a PIG-L domain deacetylase (cspld) in Chitiniphilus shinanonensis SAY3. cspld was identified using transposon mutagenesis, followed by negatively selecting a mutant incapable of growing on chitin, a polysaccharide consisting of N-acetyl-d-glucosamine (GlcNAc). We focused on the physiological role of CsPLD protein in chitin utilization., Methods and Results: Recombinant CsPLD expressed in Escherichia coli exhibited GlcNAc-6-phosphate deacetylase (GPD) activity, which is involved in the metabolism of amino sugars. However, SAY3 possesses two genes (csnagA1 and csnagA2) in its genome that code for proteins whose primary sequences are homologous to those of typical GPDs. Recombinant CsNagA1 and CsNagA2 also exhibited GPD activity with 23 and 1.6% of catalytic efficiency (kcat/Km), respectively, compared to CsPLD. The gene-disrupted mutant, Δcspld was unable to grow on chitin or GlcNAc, whereas the three mutants, ΔcsnagA1, ΔcsnagA2, and ΔcsnagA1ΔcsnagA2 grew similarly to SAY3. The determination of GPD activity in the crude extracts of each mutant revealed that CsPLD is a major enzyme that accounts for almost all cellular activities., Conclusions: Deacetylation of GlcNAc-6P catalyzed by CsPLD (but not by typical GPDs) is essential for the assimilation of chitin and its constituent monosaccharide, GlcNAc, as a carbon and energy source in C. shinanonensis., (© The Author(s) 2024. Published by Oxford University Press on behalf of Applied Microbiology International.)

Aim: This study aimed to examine foreign-educated male nurses' motivation to emigrate for work, as well as explore the contextual factors that shape their career trajectories., Background: To that end, we interviewed Chinese male nurses working in Japan. No previous study has examined why male nurses seek employment abroad or the problems they face., Methods: This study employed a qualitative and inductive research design. Sixteen Chinese male nurses working in Eastern and Western Japan were recruited using convenience and snowball sampling methods and were interviewed via video calls. Co-researchers of both genders from different occupational and cultural backgrounds coded the findings and identified common themes in participants' responses. We analysed our findings vis-à-vis various theoretical perspectives and developed an explanatory model., Results: Participants' experience as nurses in Japan ranged from six months to eight years. The findings confirmed that factors identified in earlier reports influenced nurses' motivation and work. Two new factors emerged: influence by others and social reasons . Moreover, two factors that affected male nurses' motivation to work abroad, namely, external factors and the influence of others, were identified. Social reasons were identified as a pull factor., Conclusion: Our study provides valuable insights into recruiting and retaining foreign male nurses in developed countries facing issues such as ageing populations and nursing shortages., Implications for Nursing Policy: This study has important implications for nursing management. Our findings highlight the importance of orienting foreign-educated nurses regarding the local work culture to increase the recruitment and retention of foreign talent. Moreover, enhanced salary packages and benefits to improve motivation can lead to improved job performance, which can positively impact patient and safety outcomes., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors. Published by Elsevier Ltd.)

One of the chitinases (ChiG) derived from the chitinolytic bacterium Chitiniphilus shinanonensis SAY3T exhibited chitobiase activity cleaving dimers of N-acetyl-D-glucosamine (GlcNAc) into monomers, which is not detected in typical endo-type chitinases. Analysis of the reaction products for GlcNAc hexamers revealed that all the five internal glycosidic bonds were cleaved at the initial stage. The overall reaction catalyzed by chitobiases toward GlcNAc dimers was similar to that catalyzed by N-acetyl-D-glucosaminidases (NAGs). SAY3 possesses two NAGs (ChiI and ChiT) that are thought to be important in chitin catabolism. Unexpectedly, a triple gene-disrupted mutant (ΔchiIΔchiTΔchiG) was still able to grow on synthetic medium containing GlcNAc dimers or powdered chitin, similar to the wild-type SAY3, although it exhibited only 3% of total cellular NAG activity compared to the wild-type. This indicates the presence of unidentified enzyme(s) capable of supporting normal bacterial growth on the chitin medium by NAG activity compensation., (© The Author(s) 2023. Published by Oxford University Press on behalf of Japan Society for Bioscience, Biotechnology, and Agrochemistry.)

The chitinolytic bacterium, Chitiniphilus shinanonensis SAY3 T was examined to characterize its chitin-degrading enzymes in view of its potential to convert biomass chitin into useful saccharides. A survey of the whole-genome sequence revealed 49 putative genes encoding polypeptides that are thought to be related to chitin degradation. Based on an analysis of the relative quantity of each transcript and an assay for chitin-degrading activity of recombinant proteins, a chitin degradation system driven by 19 chitinolytic enzymes was proposed. These include sixteen endo-type chitinases, two N-acetylglucosaminidases, and one lipopolysaccharide monooxygenase that catalyzes the oxidative cleavage of glycosidic bonds. Among the 16 chitinases, ChiL was characterized by its remarkable transglycosylation activity. Of the two N-acetylglucosaminidases (ChiI and ChiT), ChiI was the major enzyme, corresponding to > 98% of the total cellular activity. Surprisingly, a chiI-disrupted mutant was still able to grow on medium with powdered chitin or GlcNAc dimer. However, its growth rate was slightly lower compared to that of the wild-type SAY3. This multi-enzyme machinery composed of various types of chitinolytic enzymes may support SAY3 to efficiently utilize native chitin as a carbon and energy source and may play a role in developing an enzymatic process to decompose and utilize abundant chitin at the industrial scale., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: With the deterioration of cognitive functions, the capability to obtain information with speed, one of the essential elements needed to perform safe driving, may be impacted. We aimed to compare the legibility of analogue and digital speedometers for senior drivers with cognitive impairment, and examined the demographic, cognitive, and driving-related variables that predict their speedometer-reading performance., Methods: A total of 50 senior drivers with cognitive impairment were investigated and asked to complete an office-based speedometer-reading test using an iPad. Two general types of speedometers (analogue and digital) were tested in this study., Results: The age of the participants ranged from 61 to 92 years (mean (SD), 79.10 (6.973)), and 29 were male. The mean (SD) score of the Mini-Mental State Examination was 22.48 (6.089). The median (Q L , Q U ) scores of the analogue and digital speedometer-reading tests were 4 (4, 5.25) and 6 (6), respectively. Based on the result of the Wilcoxon signed-rank test, the score of the digital speedometer-reading test was significantly higher than that of the analogue one (Z = 4.399, P < 0.001). The results of multiple linear regression analyses show that the scores of the Mini-Mental State Examination (β = 0.358, P = 0.025), and the trail-making test-A (β = -0.443, P = 0.006) predicted the digital speedometer-reading performance, and they together explain 54.7% of the total variance., Conclusions: A digital speedometer was found to be easier for absolute value reading for senior drivers with cognitive impairment, compared to an analogue speedometer. Senior drivers with subjective cognitive decline may also have impairments in obtaining the speed information through an analogue speedometer. General cognitive function and attention may influence the speed-reading performance on the digital speedometer., (© 2022 Japanese Psychogeriatric Society.)

Background and Aim: Bacterial infection is involved in the progression of many gastrointestinal diseases, including those of pancreas; however, how and which bacteria colonize in pancreatic juice and tissue have yet to be elucidated. Recently, we reported that Enterococcus faecalis exists in the pancreatic juice and tissues of patients with chronic pancreatic disease. Here, we investigated the survival of E. faecalis in duodenal juice with different pH conditions., Methods: Pancreatic juice samples from 62 patients with cancers of the duodeno-pancreato-biliary region were evaluated for the presence of E. faecalis . 16S ribosomal RNA polymerase chain reaction and 16S-based metagenome analyses were performed to determine the bacterial composition. The survival of E. faecalis in various pancreatic juice conditions was evaluated., Results: Of 62 samples, 27% (17/62) were positive for Enterococcus spp., among which 71% (12/17) contained E. faecalis . Enterococcus spp. showed the highest fitness for survival in alkaline pancreatic juice among various bacterial species. The microbiome of pancreatic juice from patients with pancreatic and bile duct cancer showed diversity, but Enterococcus spp. were enriched among duodenal tumors and intraductal papillary mucinous neoplasms., Conclusions: Alkalinity is one of the important factors for the selective survival of E. faecalis among microbiota. E. faecalis can colonize the pancreatic duct when the pancreatic juice condition is altered., (© 2022 The Authors. JGH Open published by Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.)

Background: Time disorientation is one of the main symptoms observed in patients with dementia; however, their clock-reading ability has not been fully reported., Objective: This study aimed to investigate the clock-reading ability of both digital and analog clocks in patients with dementia. We newly devised the clock-reading test (CRT) and the number-reading test (NRT) to assess cognitive factors that may affect clock-reading ability. Furthermore, the discriminating power of the CRT was calculated., Methods: 104 participants were categorized based on their Mini-Mental State Examination (MMSE) scores as follows: subjective cognitive decline ∼ mild cognitive impairment (SCD∼MCI, N = 43), early Alzheimer's disease (AD) (N = 26), and middle-to-late AD (N = 35). Their cognitive abilities were evaluated using the clock-drawing test (CDT), CRT, and NRT., Results: Cognitive decline leads to impairment of clock-reading ability which is more pronounced in the analog clocks than digital ones. This deficit in clock-reading is attributed to a loss of semantic memory regarding clocks at all stages. Additionally, visuospatial dysfunction and reduced ability of number recognition may lead to deficit in clock-reading in the advanced stage of AD. The discriminating power of the CRT (analog) (AUC = 0.853) was high enough to detect cognitive decline., Conclusion: Digital clocks are more readable by patients with dementia. Since reading clocks is closely associated with daily life, the CRT has proved to be a useful tool. A decline of analog clock-reading may be an early detector for the onset of dementia in elderly patients.

Background: The differences in the brain activities of the insular and the visual association cortices have been reported between oral and manual stereognosis. However, these results were not conclusive because of the inherent differences in the task performance-related motor sequence conditions. We hypothesized that the involvement of the prefrontal cortex may be different between finger and oral shape discrimination. This study was conducted to clarify temporal changes in prefrontal activities occurring in the processes of oral and finger tactual shape discrimination using prefrontal functional near-infrared spectroscopy (fNIRS)., Methods: Six healthy right-handed males [aged 30.8 ± 8.2 years (mean ± SD)] were enrolled. Measurements of prefrontal activities were performed using a 22-channel fNIRS device (ETG-100, Hitachi Medical Co., Chiba, Japan) during experimental blocks that included resting state (REST), nonsense shape discrimination (SHAM), and shape discrimination (SHAPE)., Results: No significant difference was presented with regard to the number of correct answers during trials between oral and finger SHAPE discrimination. Additionally, a statistical difference for the prefrontal fNIRS activity between oral and finger shape discrimination was noted in CH 1. Finger SHAPE, as compared with SHAM, presented a temporally shifting onset and burst in the prefrontal activities from the frontopolar area (FPA) to the orbitofrontal cortex (OFC). In contrast, oral SHAPE as compared with SHAM was shown to be temporally overlapped in the onset and burst of the prefrontal activities in the dorsolateral prefrontal cortex (DLPFC)/FPA/OFC., Conclusion: The prefrontal activities temporally shifting from the FPA to the OFC during SHAPE as compared with SHAM may suggest the segregated serial prefrontal processing from the manipulation of a target image to the decision making during the process of finger shape discrimination. In contrast, the temporally overlapped prefrontal activities of the DLPFC/FPA/OFC in the oral SHAPE block may suggest the parallel procession of the repetitive involvement of generation, manipulation, and decision making in order to form a reliable representation of target objects., Competing Interests: TU and IK are employed by Dental Support Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Narita, Kamiya, Iwaki, Ishii, Endo, Shimosaka, Uchida, Kantake and Shibutani.)

Background: Prolongation of bolus forming complicates ingestion, in particular in older adults., Objectives: The purpose of this study is to examine which oral functions are associated with prolongation of the oral phase of forming a bolus until swallowing in older adults., Design: Cross-sectional study., Setting: three nursing homes in Kitakyushu, Japan from August 2017 to October 2018., Participants: 39 adults >60-years., Measurements: Number of functional teeth, chewing ability, swallowing ability, tongue and cheek pressure, saliva flow rate, oral diadochokinesis, global cognitive function, and body mass index, were examined. Time of oral phase until the first swallowing of solid food was measured as the outcome of the study using video, and audio recording of the swallowing sound by a throat microphone, with the cutoff point designated at 30 s. Based on the oral phase, participants were divided in two groups: normal and prolonged., Results: The 39 enrolled participants had a median age of 87 years, 17.3% were men, and 48.7% had prolonged oral phase. In the prolonged group, the swallowing ability, saliva flow rate, tongue and cheek pressure, and oral diadochokinesis were significantly lower than in the normal group. Binomial logistic regression analysis revealed that oral phase prolongation was associated with oral diadochokinesis (odds ratio 0.81, 95% confidence interval 0.67-0.98) after adjusting for potential covariates., Conclusion: Oral diadochokinesis deterioration is significantly associated with oral phase prolongation for initial swallowing of solid food in older adults., Competing Interests: The authors declare no conflict of interest., (© 2020 - JARLIFE.)

Wasabi (Eutrema japonicum) is a perennial plant native to Japan that is used as a spice because it contains isothiocyanates. It also contains an isosaponarin, 4'-O-glucosyl-6-C-glucosyl apigenin, in its leaves, which has received increasing attention in recent years for its bioactivity, such as its promotion of type-I collagen production. However, its biosynthetic enzymes have not been clarified. In this study, we partially purified a C-glucosyltransferase (CGT) involved in isosaponarin biosynthesis from wasabi leaves and identified the gene coding for it (WjGT1). The encoded protein was similar to UGT84 enzymes and was named UGT84A57. The recombinant enzyme of WjGT1 expressed in Escherichia coli showed C-glucosylation activity toward the 6-position of flavones such as apigenin and luteolin. The enzyme also showed significant activity toward flavonols, but trace or no activity toward flavone 4'-O-glucosides, suggesting that isosaponarin biosynthesis in wasabi plants would proceed by 6-C-glucosylation of apigenin, followed by its 4'-O-glucosylation. Interestingly, the enzyme showed no activity against sinapic acid or p-coumaric acid, which are usually the main substrates of UGT84 enzymes. The accumulation of WjGT1 transcripts was observed mainly in the leaves and flowers of wasabi, in which C-glucosylflavones were accumulated. Molecular phylogenetic analysis suggested that WjGT1 acquired C-glycosylation activity independently from other reported CGTs after the differentiation of the family Brassicaceae., (� The Author(s) 2019. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Objectives: The aims were to use different techniques to assess the degree of sensory changes and magnitude of perceived size changes in the facial region induced by nerve blocks of two different trigeminal nerve branches in healthy participants., Materials and Methods: This placebo-controlled study included 30 healthy volunteers. The participants underwent quantitative and qualitative sensory testing (QST and QualST) thrice: before, 10 min, and 2 h after mental (n = 15) and infraorbital (n = 15) nerve blocks and during control (saline) sessions. Perceived numbness, temperature changes, and perceptual distortion were also measured at all time points during the nerve block and control sessions. Differences in outcome parameters between the sessions and time points were analyzed using analyses of variance and McNemar's tests., Results: There was a significant degree of sensory loss to most QST and all QualST parameters, with significant numbness and increased perceived size at the injection site 10 min and 2 h after the nerve blocks compared with saline (P < 0.030) and the baseline (P < 0.042). Two hours after the nerve blocks, the sensitivity was significantly closer to baseline than after 10 min to most of the QST parameters (P < 0.011)., Conclusions: QST and QualST revealed that the nerve blocks in the orofacial region resulted in complete or partial blockade of afferent nerve fibers mediating thermal and mechanical function for more than 2 h with significant numbness and perceptual distortion., Clinical Relevance: Both QST and QualST can provide information on the degree of blockade of afferent nerve fibers after nerve blocks in the orofacial region.

Rutin, a 3-rutinosyl quercetin, is a representative flavonoid distributed in many plant species, and is highlighted for its therapeutic potential. In this study, we purified uridine diphosphate-rhamnose: quercetin 3-O-glucoside 6″-O-rhamnosyltransferase and isolated the corresponding cDNA (FeF3G6″RhaT) from seedlings of common buckwheat (Fagopyrum esculentum). The recombinant FeF3G6″RhaT enzyme expressed in Escherichia coli exhibited 6″-O-rhamnosylation activity against flavonol 3-O-glucoside and flavonol 3-O-galactoside as substrates, but showed only faint activity against flavonoid 7-O-glucosides. Tobacco cells expressing FeF3G6″RhaT converted the administered quercetin into rutin, suggesting that FeF3G6″RhaT can function as a rhamnosyltransferase in planta. Quantitative PCR analysis on several organs of common buckwheat revealed that accumulation of FeF3G6″RhaT began during the early developmental stages of rutin-accumulating organs, such as flowers, leaves, and cotyledons. These results suggest that FeF3G6″RhaT is involved in rutin biosynthesis in common buckwheat.

Objective: Aspiration of subglottic secretion is a widely used intervention to prevent ventilator-associated pneumonia (VAP). This study aimed to compare the efficacy of continuous and intermittent subglottic secretion drainage (SSD) in preventing VAP., Methods: A single-center randomized controlled trial was conducted on adult postoperative patients who were expected to undergo mechanical ventilation for more than 48 hours. Primary outcome measure was incidence of VAP and secondary outcome measures were length of mechanical ventilation and intensive-care unit (ICU) stay., Results: Fifty-nine patients received continuous SSD, while 60 patients received intermittent SSD. Of these 119 patients, 88 (74%) were excluded and 15 and 16 patients were allocated to receive continuous and intermittent SSD, respectively. VAP was detected in 4 (26.7%) and 7 (43.8%) patients in the continuous and intermittent groups, respectively, (p=0.320). The length of mechanical ventilation was significantly shorter (p=0.034) in the continuous group (99.5±47.1 h) than in the intermittent group (159.9±94.5 h). The length of ICU stay was also shorter (p=0.0097) in the continuous group (6.3±2.1 days) than the intermittent group (9.8±4.8 days)., Conclusions: Although continuous SSD did not reduce the incidence of VAP, it reduced the length of mechanical ventilation and ICU stay when compared to intermittent SSD., Competing Interests: CONFLICTS OF INTEREST We have nothing to declare for this study.

Citrus plants accumulate many kinds of flavonoids, including di-C-glucosyl flavonoids, which have attracted considerable attention due to their health benefits. However, the biosynthesis of di-C-glucosyl flavonoids has not been elucidated at the molecular level. Here, we identified the C-glycosyltransferases (CGTs) FcCGT (UGT708G1) and CuCGT (UGT708G2) as the primary enzymes involved in the biosynthesis of di-C-glucosyl flavonoids in the citrus plants kumquat (Fortunella crassifolia) and satsuma mandarin (Citrus unshiu), respectively. The amino acid sequences of these CGTs were 98% identical, indicating that CGT genes are highly conserved in the citrus family. The recombinant enzymes FcCGT and CuCGT utilized 2-hydroxyflavanones, dihydrochalcone, and their mono-C-glucosides as sugar acceptors and produced corresponding di-C-glucosides. The K m and k cat values of FcCGT toward phloretin were <0.5 μm and 12.0 sec -1 , and those toward nothofagin (3'-C-glucosylphloretin) were 14.4 μm and 5.3 sec -1 , respectively; these values are comparable with those of other glycosyltransferases reported to date. Transcripts of both CGT genes were found to concentrate in various plant organs, and particularly in leaves. Our results suggest that di-C-glucosyl flavonoid biosynthesis proceeds via a single enzyme using either 2-hydroxyflavanones or phloretin as a substrate in citrus plants. In addition, Escherichia coli cells expressing CGT genes were found to be capable of producing di-C-glucosyl flavonoids, which is promising for commercial production of these valuable compounds., (© 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.)

Currently, the immediate extubation in the operating room is necessary for the patients undergoing off-pump coronary artery bypass (OPCAB). To achieve UFTA, anesthesiologists should control general anesthesia. We report the case series of UFTA for the patients undergone OPCAB at our hospital. Forty patients receiving OPCAB between April 2012 and April 2014 were retrospectively analyzed. We divided the patients into those extubated in the operating room (Extubation group) and those not (Intubation group). The extubation criteria included adequate revascularization, adequate hemostasis, ratio between Pa(O2) and FI(O2) more than 250, body temperature above 36 degrees C, stable hemodynamics without noradrenaline, without IABP, and normal postoperative chest X-ray. Twenty-three patients (70% of the scheduled and 20% of the emergency patients) were included in our criteria and could be extubated in the operating room. One patient was re-intubated due to asthma attack following extubation in the operating room. The sucess rate of immediate extubation was 95.7% by our criteria. There were no significant differences in age, sex, BMI, duration of anesthesia, and amount of hemorrhage between the 2 groups. We believe that UFTA for OPCAB patients may be possible under carefull decision by extubation criteria.

Osteoclasts are bone-specific multinucleated cells generated by the differentiation of monocyte/macrophage lineage precursors. Regulation of osteoclast differentiation is considered an effective therapeutic approach to the treatment of bone-lytic diseases. Periodontitis is an inflammatory disease characterized by extensive bone resorption. In this study, we investigated the effects of sodium fluoride (NaF) on osteoclastogenesis induced by Porphyromonas gingivalis, an important colonizer of the oral cavity that has been implicated in periodontitis. NaF strongly inhibited the P. gingivalis-induced alveolar bone loss. That effect was accompanied by decreased levels of cathepsin K, interleukin (IL)-1β, matrix metalloproteinase 9 (MMP9), and tartrate-resistant acid phosphatase, which were up-regulated during P. gingivalis-induced osteoclastogenesis. Consistent with the in vivo anti-osteoclastogenic effect, NaF inhibited osteoclast formation caused by the differentiation factor RANKL (receptor activator of nuclear factor κB ligand) and macrophage colony-stimulating factor (M-CSF). The RANKL-stimulated induction of the transcription factor nuclear factor of activated T cells (NFAT) c1 was also abrogated by NaF. Taken together, our data demonstrate that NaF inhibits RANKL-induced osteoclastogenesis by reducing the induction of NFATc1, ultimately leading to the suppressed expression of cathepsin K and MMP9. The in vivo effect of NaF on the inhibition of P. gingivalis-induced osteoclastogenesis strengthens the potential usefulness of NaF for treating periodontal diseases.

Chitin, a linear polysaccharide consisting of β-1,4-linked N-acetyl-D-glucosamine (GlcNAc), is widely used because of its biochemical properties. GlcNAc oligomers prepared from chitin have useful biological activities, such as immunostimulation and the induction of plant defence responses. Microbial chitinolytic enzymes have been investigated extensively for their potential use in the eco-friendly enzymatic production of GlcNAc and its oligomers. Chitiniphilus shinanonensis SAY3(T) is a recently found bacterium with a strong chitinolytic activity. The chitinolytic enzymes from this strain are potentially useful for the efficient production of GlcNAc and its oligomers from chitin. ChiL from C. shinanonensis is an endo-type chitinase belonging to the family 18 glycoside hydrolases (GH18). To understand the enzymatic reaction mechanism of ChiL and utilize it for further enzyme engineering, the catalytic domain (41-406) of ChiL, the construct for which was carefully designed, was expressed, purified and crystallized by the vapour-diffusion method. The crystal belonged to the orthorhombic space group P212121, with unit-cell parameters a = 69.19, b = 81.55, c = 130.01 Å, and diffracted to 1.25 Å resolution. The Matthews coefficient (VM = 2.2 Å(3) Da(-1)) suggested the presence of two monomers per asymmetric unit with a solvent content of 45%.

The aim of this study was to evaluate the relationship between pain intensities and psychosocial characteristics in middle-aged and older patients with temporomandibular disorders (TMDs) and burning mouth syndrome (BMS). Subjects were selected according to the Research Diagnostic Criteria for TMD (n = 705) and International Association for the Study of Pain criteria for BMS (n = 175). Patients were then divided into two age groups: 45-64 years (middle-aged, Group A) and 65-84 years (older, Group B). Pain intensity and depression and somatization scores were evaluated in both groups. In BMS patients, present and worst pain intensities were significantly higher in Group B than in Group A {4.6 [95% confidence interval (CI) = 4.0-5.2] vs. 3.5 [95% CI = 3.1-3.9] and 5.9 [95% CI = 5.2-6.4] vs. 5.0 [95% CI = 4.5-5.6], respectively; P < 0.05}, with no difference observed in TMD patients. The depression and somatization scores were significantly higher in Group A than in Group B among BMS patients [0.57 (95% CI = 0.45-0.69) vs. 0.46 (95% CI = 0.34-0.59) and 0.537 (95% CI = 0.45-0.63) vs. 0.45 (95% CI = 0.34-0.55); P < 0.05], with no difference observed in TMD patients. The results of the present study indicate that pain intensities and psychosocial characteristics in BMS appear to differ between middle-aged and older patients.

C-Glycosides are characterized by their C-C bonds in which the anomeric carbon of the sugar moieties is directly bound to the carbon atom of aglycon. C-Glycosides are remarkably stable, as their C-C bonds are resistant to glycosidase or acid hydrolysis. A variety of plant species are known to accumulate C-glycosylflavonoids; however, the genes encoding for enzymes that catalyze C-glycosylation of flavonoids have been identified only from Oryza sativa (rice) and Zea mays (maize), and have not been identified from dicot plants. In this study, we identified the C-glucosyltransferase gene from the dicot plant Fagopyrum esculentum M. (buckwheat). We purified two isozymes from buckwheat seedlings that catalyze C-glucosylation of 2-hydroxyflavanones, which are expressed specifically in the cotyledon during seed germination. Following purification we isolated the cDNA corresponding to each isozyme [FeCGTa (UGT708C1) and FeCGTb (UGT708C2)]. When expressed in Escherichia coli, both proteins demonstrated C-glucosylation activity towards 2-hydroxyflavanones, dihydrochalcone, trihydroxyacetophenones and other related compounds with chemical structures similar to 2',4',6'-trihydroxyacetophenone. Molecular phylogenetic analysis of plant glycosyltransferases shows that flavonoid C-glycosyltransferases form a different clade with other functionally analyzed plant glycosyltransferases., (© 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.)

We investigated the relationship between pain intensity and psychosocial characteristics in patients with temporomandibular disorder (TMD). Participants with painful TMD, according to the Research Diagnostic Criteria for TMD (RDC/TMD), were recruited from our clinic and classified into six age groups: 15 to 24, 25 to 34, 35 to 44, 45 to 54, 55 to 64, and 65 to 85 years (Groups A through F, respectively). Self-reported present pain intensity and worst pain intensity during the past 6 months were ascertained using a numeric rating scale (0 to 10). Depression and somatization scores were evaluated using the RDC/TMD axis II questionnaire. Among women, worst pain intensity was significantly lower in Groups E and F than in Groups B and C (P < 0.05). In Groups A, C, and D, depression scores were significantly higher in women than in men (P < 0.05). Among women, depression score was significantly lower in Group F than in Groups A through C (P < 0.05). In Groups A through D, somatization scores were significantly higher for women than for men (P < 0.05). Depression score and TMD symptom severity appear to decrease with age in women.

A full-length cDNA coding for a putative adenosine deaminase (Fv-ada) was isolated from the basidiomycete Flammulina velutipes. Fv-ada encodes a polypeptide consisting of 537 amino acid residues, which has a consensus sequence conserved among adenosine deaminase-related growth factors (ADGF) found in several metazoa, including chordates and insects. Fv-ada transcript was detected at all stages of growth in dikaryotic F. velutipes cells, with a peak at the primordial stage. Heterologous expression of Fv-ada in the yeast Pichia pastoris produced recombinant Fv-ADA that catalyzed the conversion of adenosine to inosine. Dikaryotic mycelia from F. velutipes were transformed with the binary plasmid pFungiway-Fv-ada, which was designed to suppress the expression of Fv-ada through RNA interference. The growth rates of the resulting transformants were retarded in response to the degree of suppression, indicating that Fv-ada plays an important role in the mycelial growth of F. velutipes. These results suggested that ADGF could function as growth factors in fungi, as is seen in other eukaryotes., (Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Background: The inhibition of transforming growth factor β (TGF-β)-induced myofibroblast differentiation is a key objective for the treatment of hypertrophic scarring. We previously reported that knockdown of the electron transfer flavoprotein β subunit (ETFB) reduced mechanoregulated cell number in fibroblast-populated collagen gel cultures [1]., Objective: To characterize the effects of ETFB knockdown, we investigated gel contraction, TGF-β-induced collagen, α-SMA mRNA expression and stress fiber formation., Methods: Fibroblasts were transfected with negative control or ETFB-specific siRNAs and embedded in collagen gels in an attached or detached condition. The gel contraction assay was performed in three different concentrations of collagen (0.5, 1.0 or 1.5mg/mL) and was analyzed by measuring the changes in the gel area throughout the culture period. The attached collagen gel culture was performed in the presence of rTGF-β and the mRNA levels of α-SMA and COL1A1 were measured by qRT-PCR. The effect of ETFB knockdown on proliferation and stress fiber organization in monolayer cultures was investigated by conducting AlamarBlue assays and phalloidin staining., Results: The transfection of ETFB siRNA did not alter gel contraction compared to the negative control in all collagen concentrations. When the cells were treated with TGF-β under mechanical stress conditions, ETFB knockdown attenuated α-SMA mRNA expression to a level comparable to that observed in the absence of TGF-β. However, no inhibitory effect on COL1A1 mRNA levels was observed. The AlamarBlue assay indicated that the knockdown had no effect on the proliferation of cells cultured on plastic. Phalloidin staining of a monolayer culture showed that ETFB knockdown weakened the stress fiber organization induced by rTGF-β., Conclusion: ETFB knockdown can affect TGF-β-induced tissue remodeling and/or fibrotic processes in vitro., (Copyright © 2012 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.)

Chitiniphilus shinanonensis type strain SAY3(T) is a strongly chitinolytic bacterium, originally isolated from the moat water in Ueda, Japan. To elucidate the chitinolytic activity of this strain, 15 genes (chiA-chiO) coding for putative chitin-degrading enzymes were isolated from a genomic library. Sequence analysis revealed the genes comprised 12 family 18 chitinases, a family 19 chitinase, a family 20 β-N-acetylglucosaminidase, and a polypeptide with a chitin-binding domain but devoid of a catalytic domain. Two operons were detected among the sequences: chiCDEFG and chiLM. The gene coding for the polypeptide (chiN) showed sequence similarity to family 19 chitinases and was successfully expressed in Escherichia coli. ChiN demonstrated a multi-domain structure, composed of the N-terminal, two chitin-binding domains connected by a Pro- and Thr-rich linker, and a family 19 catalytic domain located at the C-terminus. The recombinant protein rChiN catalyzed an endo-type cleavage of N-acetyl-d-glucosamine oligomers, and also degraded insoluble chitin and soluble chitosan (degree of deacetylation of 80%). rChiN exhibited an inhibitory effect on hyphal growth of the fungus Trichoderma reesei. The chitin-binding domains of ChiN likely play an important role in the degradation of insoluble chitin, and are responsible for a growth inhibitory effect on fungi., (Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

This study compared pain intensity and psychosocial characteristics between patients with burning mouth syndrome (BMS) and those with trigeminal neuralgia (TN). Data from 282 patients with BMS and 83 patients with TN were analyzed. Patients reported duration of illness: duration ≤ 6 months was defined as acute illness and > 6 months as chronic illness. Present pain intensity and worst pain intensity during the past 6 months were reported using a 0-10 numeric rating scale (NRS). In addition, depression and somatization scores were evaluated on questionnaires. Patients with chronic BMS reported significantly higher pain intensity and had worse psychosocial characteristics than did those with acute BMS. Pain intensity was higher in TN patients than in BMS patients, although neither pain intensity nor psychosocial characteristics significantly differed between patients with acute and chronic illness. Logistic regression analysis of BMS and TN patients revealed that the odds ratio for worst pain was significantly lower for BMS patients than for TN patients and that the odds ratio for somatization score was 3.8 times higher in BMS patients. These findings suggest that BMS patients may require pain control targeting the central nervous system or psychosocial characteristics.

Chitiniphilus shinanonensis strain SAY3(T) is a chitinolytic bacterium isolated from moat water of Ueda Castle in Nagano Prefecture, Japan. Fifteen genes encoding putative chitinolytic enzymes (chiA-chiO) have been isolated from this bacterium. Five of these constitute a single operon (chiCDEFG). The open reading frames of chiC, chiD, chiE, and chiG show sequence similarity to family 18 chitinases, while chiF encodes a polypeptide with two chitin-binding domains but no catalytic domain. Each of the five genes was successfully expressed in Escherichia coli, and the resulting recombinant proteins were characterized. Four of the recombinant proteins (ChiC, ChiD, ChiE, and ChiG) exhibited endo-type chitinase activity toward chitinous substrates, while ChiF showed no chitinolytic activity. In contrast to most endo-type chitinases, which mainly produce a dimer of N-acetyl-D-glucosamine (GlcNAc) as final product, ChiG completely split the GlcNAc dimer into GlcNAc monomers, indicating that it is a novel chitinase.

A gene coding for a major extracellular chitosanase was isolated from Aspergillus oryzae IAM2660. It had a multi-domain structure composed of a signal peptide, a catalytic domain, Thr- and Pro-rich linkers, and repeated peptides (the R3 domain) from the N-terminus. The R3 domain bound to insoluble powder chitosan, but it did not promote the hydrolysis rate of the chitosanase to any extent.

Background: Fibroblast activation is strongly influenced by mechanical environment in the wound-healing process, especially in fibrosis. Mechanically stressed three-dimensional collagen embedded culture is a useful model representing fibroblasts in morphological as well as biochemical situations encountered during fibrosis., Objective: To find key proteins involved in reducing the number of fibroblasts during mechanical stress, we performed two-dimensional gel electrophoresis (2DE)-based differential display and siRNA-based functional screening with collagen gel culture focusing on the differences between attached and detached culture environments., Methods: Membrane extracts of fibroblasts from 1 day of attached or detached cultures were subjected to 2DE. We compared protein expression levels and identified the attached-culture-dominant proteins by MALDI-TOF-MS. Next, fibroblasts were transfected with siRNA and embedded in collagen gel. Cell number was counted after 3 days in culture., Results: Eight attached culture dominant proteins were identified with MALDI-TOF-MS. Transfection of siRNA against these proteins demonstrated that electron transfer flavoprotein β subunit (ETFB)-specific siRNA reduced the cell number in the attached culture without a decrease in the detached culture., Conclusion: ETFB participates in the mechanoregulation of fibroblast cell number in collagen gel culture., (Copyright © 2011 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.)

In general anesthesia for a patient with dystrophia myotonica (DM), respiratory depression and muscle weakness by opioid, as well as prolongation of the effect of muscle relaxant are seen postoperatively. Therefore it is desirable to choose agents with short duration of action and to dose these medicines to the minimum. We report a case of a 45-year-old woman with DM who underwent laparotomy for uterine cancer under general anesthesia combined with epidural anesthesia. Epidural catheter was placed from T 11-12, and anesthesia was inducted with propofol and remifentanil (RF). We administered rocuronium bromide (RB) 5 mg while watching TOF ratio with a muscle relaxation monitor (TOF-Watch). T1 became 0 after giving a dose of 10 mg, and intubation was performed. We maintained anesthesia by propofol and RF combined with epidural anesthesia. TOF ratio was restored to around 80% 90 minutes after RB administration, but we did not give supplemental doses because the operation went well smoothly. Recovery was smooth and fast. The respiratory depression and the muscle spasm were not noticed. RB and RE both with short duration of action, are useful in anesthesia management in DM cases.

The temporal changes of a bacterial community in soil with chitin or chitosan added were analyzed by PCR-denaturing gradient gel electrophoresis (DGGE) targeting the 16S rRNA gene using total DNAs prepared from the community. Band patterns of PCR-DGGE confirmed that 31 species become predominant after the addition of chitin or chitosan. The determination of the nucleotide sequences of the bands of the 31 species indicated that 20 species belonged to the division Proteobacteria, and that the genus Cellvibrio was apparently predominant among them (7/20). The 16S rRNA sequences of the 16 deduced species (16/31) showed less than 98% similarities to those of previously identified bacteria, indicating that the species were derived from unidentified bacteria. The total community DNAs extracted from bacterial cells adsorbed on the surface of flakes of chitin and chitosan placed in a river, a moat, or soil were subjected to PCR-DGGE to examine the extent of diversity of chitinolytic bacteria among different environments. The predominant species significantly differed between the chitin and chitosan placed in the river and moat, but not so much between those placed in the soil. The large difference between the diversities of the three bacterial communities indicated that a wide variety of bacteria including unidentified ones are involved in the degradation of chitin and chitosan in the above-mentioned natural environments., ((c) 2009 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Agrobacterium tumefaciens was used to transform the vegetative dikaryotic mycelium of Flammulina velutipes using a hygromycin B resistance gene as selectable marker. The gene coding for urogen III methyltransferase (cob) was introduced into F. velutipes dikaryotic cells. The resulting transformant cells generated a bright red fluorescence, indicating that cob is promising as a reporter gene in F. velutipes.

A stable bacterial community expressing strong chitinolytic activity was constructed by mixing and cultivating chitinolytic bacteria collected from different natural sources. The DNA fragment pattern, after PCR-denaturing gradient gel electrophoresis (DGGE) targeting 16S rRNA genes using total DNA prepared from whole cells, indicated that the community was composed of four dominant bacterial species. All four species were isolated on agar medium, and one strain, SAY3, was deduced to be a novel species belonging to a new genus based on the 16S rRNA nucleotide sequence. The other strains showed high similarity in their 16S rRNA sequences to those of previously identified bacteria (Acinetobacter and Microbacterium). Strain SAY3 degraded and utilized larger particles of chitin faster than the community, indicating that it plays an important role in the chitin degradation conferred by the community.

The filamentous fungus Coleophoma empetri F-11899 produces an echinocandin-like compound FR901379, the original source for micafungin which is prescribed to treat deep-seated mycoses. Despite its industrial importance, no genetic information on C. empetri F-11899 is currently available. To characterize FR901379 biosynthetic genes by insertional mutagenesis and to improve the compound production genetically, Agrobacterium tumefaciens-mediated transformation (ATMT) was attempted to make genetic manipulation possible in this strain. The optimum conditions for ATMT of C. empetri were determined for the cell density of bacteria, time period of co-cultivation and types of filters in co-cultivation. Using the established ATMT method, the hygromycin B resistant gene was successfully transferred into the genome of C. empetri F-11899 and stably maintained even after a serial passage. Some of these results will be applicable for ATMT of various filamentous fungi.