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Background: Corneal dystrophies (CDs) significantly affect quality of life. However, their progression and characteristics remain unclear. This study aimed to report a case of a unilateral variant of lattice corneal dystrophy (LCD) with c.1501C>A (p.P501T) and c.1733T>C (p.L578P) variants in the transforming growth factor-beta-induced (TGFBI) gene., Case Presentation: A 39-year-old Japanese woman presented with ocular pain and decreased visual acuity in the left eye. A slit-lamp examination of her left cornea revealed recurrent corneal erosion complicated by contact lens-associated infectious keratitis, fine lattice lines, and central corneal haze in the anterior stroma, with no opacities in the right cornea. In vivo confocal microscopic examination of the right eye showed highly reflective branching filaments in the corneal stroma, whereas the left cornea was unremarkable. Based on these clinical findings, we diagnosed the patient with unilateral LCD. The molecular genetic analysis revealed the TGFBI: a c.1501C>A (p.P501T) variant in exon 11 and the c.1733T>C (p.L578P) variant in exon 13., Conclusion: A 39-year-old female patient with LCD with c.1501C>A (p.P501T) and c.1733T>C (p.L578P) TGFBI variants exhibited unilateral corneal findings, including recurrent corneal erosion, fine lattice lines, and central corneal haze in the anterior stroma. The study's findings could benefit CD treatment.

Heterochromatin is a nuclear area that contains highly condensed and transcriptionally inactive chromatin. Alterations in the organization of heterochromatin are correlated with changes in gene expression and genome stability, which affect various aspects of plant life. Thus, studies of the molecular mechanisms that regulate heterochromatin organization are important for understanding the regulation of plant physiology. Microscopically, heterochromatin can be characterized as chromocenters that are intensely stained with DNA-binding fluorescent dyes. Arabidopsis thaliana exhibits distinctive chromocenters in interphase nuclei, and genetic studies combined with cytological analyses have identified a number of factors that are involved in heterochromatin assembly and organization. In this review, I will summarize the factors involved in the regulation of heterochromatin organization in plants., (© 2024. The Author(s) under exclusive licence to The Botanical Society of Japan.)

Objectives: To develop the Parenting Behavior Checklist to Promote Preschoolers' sleep (PCPP), quantify sleep-promoting parenting behaviors for children, and examine the scale's reliability and validity., Methods: The PCPP was developed based on the recommendations of the ABCs of SLEEPING for children's sleep, which is strongly supported by research evidence. Its validity and reliability were evaluated using data from 140 participants. Structural validity was estimated using exploratory factor analysis (EFA) and confirmatory factor analysis (CFA), and internal consistency was evaluated by Cronbach's α. Hypothesis testing was evaluated by analyzing the correlations between each factor of the Japanese Sleep Questionnaire for Preschoolers (JSQ-P) and the PCPP., Results: Regarding structural validity, EFA was conducted because CFA showed a poor model fit. The PCPP comprised one factor and six items. The JSQ-P subfactors of insomnia or circadian rhythm sleep-wake disorders, undesirable morning symptoms and behaviors, and insufficient sleep were moderately negatively correlated with the PCPP; the subfactor of undesirable daytime behaviors related to sleep problems was weakly negatively correlated with the PCPP. Thus, the sleep-promoting parenting behaviors listed in the PCPP were associated with better sleep in children., Conclusions: The PCPP showed sufficient reliability and validity. Future studies should use the scale to examine more effective interventions regarding sleep-promoting parental behaviors for children.

Although oxytocin may provide a novel therapeutics for the core features of autism spectrum disorder (ASD), previous results regarding the efficacy of repeated or higher dose oxytocin are controversial, and the underlying mechanisms remain unclear. The current study is aimed to clarify whether repeated oxytocin alter plasma cytokine levels in relation to clinical changes of autism social core feature. Here we analyzed cytokine concentrations using comprehensive proteomics of plasmas of 207 adult males with high-functioning ASD collected from two independent multi-center large-scale randomized controlled trials (RCTs): Testing effects of 4-week intranasal administrations of TTA-121 (A novel oxytocin spray with enhanced bioavailability: 3U, 6U, 10U, or 20U/day) and placebo in the crossover discovery RCT; 48U/day Syntocinon or placebo in the parallel-group verification RCT. Among the successfully quantified 17 cytokines, 4 weeks TTA-121 6U (the peak dose for clinical effects) significantly elevated IL-7 (9.74, 95 % confidence interval [CI] 3.59 to 15.90, False discovery rate corrected P (P FDR ) < 0.001), IL-9 (56.64, 20.46 to 92.82, P FDR  < 0.001) and MIP-1b (18.27, 4.96 to 31.57, P FDR  < 0.001) compared with placebo. Inverted U-shape dose-response relationships peaking at TTA-121 6U were consistently observed for all these cytokines (IL-7: P < 0.001; IL-9: P < 0.001; MIP-1b: P = 0.002). Increased IL-7 and IL-9 in participants with ASD after 4 weeks TTA-121 6U administration compared with placebo was verified in the confirmatory analyses in the dataset before crossover (P FDR  < 0.001). Furthermore, the changes in all these cytokines during 4 weeks of TTA-121 10U administration revealed associations with changes in reciprocity score, the original primary outcome, observed during the same period (IL-7: Coefficient = -0.05, -0.10 to 0.003, P = 0.067; IL-9: -0.01, -0.02 to -0.003, P = 0.005; MIP-1b: -0.02, -0.04 to -0.007, P = 0.005). These findings provide the first evidence for a role of interaction between oxytocin and neuroinflammation in the change of ASD core social features, and support the potential role of this interaction as a novel therapeutic seed. Trial registration: UMIN000015264, NCT03466671/UMIN000031412., Competing Interests: Declaration of Competing Interest 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., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Kidney transplantation is a common yet highly demanding medical procedure worldwide, enhancing the quality of life for patients with chronic kidney disease. Despite its prevalence, the procedure faces a shortage of available organs, partly due to contamination by microorganisms, leading to significant organ disposal. This study proposes utilizing photonic techniques associated with organ support machines to prevent patient contamination during kidney transplantation. We implemented a decontamination system using ultraviolet-C (UV-C) irradiation on the preservation solution circulating through pigs' kidneys between harvest and implant. UV-C irradiation, alone or combined with ultrasound (US) and Ps80 detergent during ex-vivo swine organ perfusion in a Lifeport® Kidney Transporter machine, aimed to reduce microbiological load in both fluid and organ. Results show rapid fluid decontamination compared to microorganism release from the organ, with notable retention. By including Ps80 detergent at 0.5% during UV-C irradiation 3 log 10 (CFU mL -1 ) of Staphylococcus aureus bacteria previously retained in the organ were successfully removed, indicating the technique's feasibility and effectiveness., (© 2024. The Author(s).)

Background: Given the global shortage of child psychiatrists and barriers to specialized care, remote assessment is a promising alternative for diagnosing and managing attention-deficit/hyperactivity disorder (ADHD). However, only a few studies have validated the accuracy and acceptability of these remote methods., Objective: This study aimed to test the agreement between remote and face-to-face assessments., Methods: Patients aged between 6 and 17 years with confirmed Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition diagnoses of ADHD or autism spectrum disorder (ASD) were recruited from multiple institutions. In a randomized order, participants underwent 2 evaluations, face-to-face and remotely, with distinct evaluators administering the ADHD Rating Scale-IV (ADHD-RS-IV). Intraclass correlation coefficient (ICC) was used to assess the reliability of face-to-face and remote assessments., Results: The participants included 74 Japanese children aged between 6 and 16 years who were primarily diagnosed with ADHD (43/74, 58%) or ASD (31/74, 42%). A total of 22 (30%) children were diagnosed with both conditions. The ADHD-RS-IV ICCs between face-to-face and remote assessments showed "substantial" agreement in the total ADHD-RS-IV score (ICC=0.769, 95% CI 0.654-0.849; P<.001) according to the Landis and Koch criteria. The ICC in patients with ADHD showed "almost perfect" agreement (ICC=0.816, 95% CI 0.683-0.897; P<.001), whereas in patients with ASD, it showed "substantial" agreement (ICC=0.674, 95% CI 0.420-0.831; P<.001), indicating the high reliability of both methods across both conditions., Conclusions: Our study validated the feasibility and reliability of remote ADHD testing, which has potential benefits such as reduced hospital visits and time-saving effects. Our results highlight the potential of telemedicine in resource-limited areas, clinical trials, and treatment evaluations, necessitating further studies to explore its broader application., Trial Registration: UMIN Clinical Trials Registry UMIN000039860; http://tinyurl.com/yp34x6kh., (©Shunya Kurokawa, Kensuke Nomura, Nana Hosogane, Takashi Nagasawa, Yuko Kawade, Yu Matsumoto, Shuichi Morinaga, Yuriko Kaise, Ayana Higuchi, Akiko Goto, Naoko Inada, Masaki Kodaira, Taishiro Kishimoto. Originally published in the Journal of Medical Internet Research (https://www.jmir.org), 19.02.2024.)

ACTIN DEPOLYMERIZING FACTOR (ADF) is a conserved protein that regulates the organization and dynamics of actin microfilaments. Eleven ADFs in the Arabidopsis thaliana genome are grouped into four subclasses, and subclass I ADFs, ADF1-4, are all expressed throughout the plant. Previously, we showed that subclass I ADFs function in the regulation of the response against powdery mildew fungus as well as in the regulation of cell size and endoreplication. Here, we report a new role of subclass I ADFs in the regulation of nuclear organization and gene expression. Through microscopic observation of epidermal cells in mature leaves, we found that the size of chromocenters in both adf4 and transgenic lines where expression of subclass I ADFs is downregulated (ADF1-4Ri) was reduced compared with that of wild-type Col-0. Arabidopsis thaliana possesses eight ACTIN (ACT) genes, among which ACT2, -7 and -8 are expressed in vegetative organs. The chromocenter size in act7, but not in the act2/8 double mutant, was enlarged compared with that in Col-0. Microarray analysis revealed that 1,818 genes were differentially expressed in adf4 and ADF1-4Ri. In particular, expression of 22 nucleotide-binding leucine-rich repeat genes, which are involved in effector-triggered plant immunity, was reduced in adf4 and ADF1-4Ri. qRT-PCR confirmed the altered expressions shown with microarray analysis. Overall, these results suggest that ADF regulates various aspects of plant physiology through its role in regulation of nuclear organization and gene expression. The mechanism how ADF and ACT regulate nuclear organization and gene expression is discussed., (© The Author(s) 2023. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Carbon dots (CDs) correspond to carbon-based materials (CBM) with sizes usually below 10 nm. These nanomaterials exhibit attractive properties such us low toxicity, good stability, and high conductivity, which have promoted their thorough study over the past two decades. The current review describes four types of CDs: carbon quantum dots (CQDs), graphene quantum dots (GQDs), carbon nanodots (CNDs), and carbonized polymers dots (CPDs), together with the state of the art of the main routes for their preparation, either by "top-down" or "bottom-up" approaches. Moreover, among the various usages of CDs within biomedicine, we have focused on their application as a novel class of broad-spectrum antibacterial agents, concretely, owing their photoactivation capability that triggers an enhanced antibacterial property. Our work presents the recent advances in this field addressing CDs, their composites and hybrids, applied as photosensitizers (PS), and photothermal agents (PA) within antibacterial strategies such as photodynamic therapy (PDT), photothermal therapy (PTT), and synchronic PDT/PTT. Furthermore, we discuss the prospects for the possible future development of large-scale preparation of CDs, and the potential for these nanomaterials to be employed in applications to combat other pathogens harmful to human health. This article is categorized under: Therapeutic Approaches and Drug Discovery > Nanomedicine for Infectious Disease., (© 2023 Wiley Periodicals LLC.)

Background: A microfluidic real-time polymerase chain reaction (PCR) system can rapidly detect the viral DNA in specimens. Detection of herpes simplex virus (HSV) and varicella-zoster virus (VZV) DNA in tears is a useful diagnostic tool for herpes simplex virus keratitis (HSK) and herpes zoster ophthalmicus (HZO)., Methods: In total, 20 patients were included in this cross-sectional study. Among them, 8 patients with infectious epithelial HSK and 12 patients with HZO were included in HSK and HZO groups, respectively. In addition, 8 patients with non-herpetic keratitis and 4 healthy individuals without keratitis were included in the control group. Numbers of HSV and VZV DNA copies in tears of all patients and individuals were evaluated using a microfluidic real-time PCR system. Regarding HSV/VZV DNA test, tear specimens were collected by filter paper method using Schirmer's test paper, and subsequently, DNA was extracted from the filter paper using an automated nucleic acid extractor. Afterward, quantitative PCR was performed using a microfluidic real-time PCR system., Results: From tear collection to real-time PCR result determination, the HSV/VZV DNA test took approximately 40 min. In the HSK group, the sensitivity and specificity of the HSV DNA tests were 100% each. The median value (range) of number of HSV DNA copies for affected eyes was 3.4 × 10 5 copies/μL (under a lower detection limit of 7.6). In the HZO group, the sensitivity and specificity of the VZV DNA tests were 100% each. The median value (range) of number of VZV DNA copies for affected eyes was 5.3 × 10 5 copies/μL (under a lower detection limit of 5.6 × 10 -2 )., Conclusion: In conclusion, quantitative PCR for HSV and VZV DNA in tears using a microfluidic real-time PCR system is useful for diagnosing and monitoring HSK and HZO., (© 2023. The Author(s).)

All aspects of plant physiology are influenced by temperature. Changes in environmental temperature alter the temperatures of plant tissues and cells, which then affect various cellular activities, such as gene expression, protein stability and enzyme activities. In turn, changes in cellular activities, which are associated with either exothermic or endothermic reactions, can change the local temperature in cells and tissues. In the past 10 years, a number of fluorescent probes that detect temperature and enable intracellular temperature imaging have been reported. Intracellular temperature imaging has revealed that there is a temperature difference >1°C inside cells and that the treatment of cells with mitochondrial uncoupler or ionomycin can cause more than a 1°C intracellular temperature increase in mammalian cultured cells. Thermogenesis mechanisms in brown adipocytes have been revealed with the aid of intracellular temperature imaging. While there have been no reports on plant intracellular temperature imaging thus far, intracellular temperature imaging is expected to provide a new way to analyze the mechanisms underlying the various activities of plant cells. In this review, I will first summarize the recent progress in the development of fluorescent thermometers and their biological applications. I will then discuss the selection of fluorescent thermometers and experimental setup for the adaptation of intracellular temperature imaging to plant cells. Finally, possible applications of intracellular temperature imaging to investigate plant cell functions will be discussed., (© The Author(s) 2022. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Brown adipose tissue specializes in expending energy through non-shivering thermogenesis, and many studies have associated its activity with protection and treatment of obesity and metabolic diseases. To reveal the mechanisms involved in heat production, primary cultured brown adipose cells (BACs) have been used because of their ease of genetic engineering and similarity to living tissue. However, thermogenic activity has often been evaluated as an indirect method, such as the measurement of oxygen consumption. Recently, fluorescent nanothermometers for the direct measurement of intracellular temperature have been developed and applied to elucidate the mechanisms of heat production in BACs. In this chapter, we introduce a protocol that uses a cationic fluorescent polymeric thermometer to directly measure the temperature within primary cultured BACs. We anticipate that this protocol will be beneficial in elucidating the mechanism of thermogenesis in BACs., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Dupilumab-induced ocular surface disease (DIOSD) has been reported in patients with atopic dermatitis treated with dupilumab, and has been recognized as an adverse event of dupilumab. Our objective was to describe two cases of DIOSD with alterations in eotaxin-2 and interleukin (IL)-8 messenger ribonucleic acid (mRNA) expression on the ocular surface., Case Presentation: In the ocular surface test, specimens were collected from the patient's ocular surface, and eotaxin-2 and IL-8 mRNA levels in the specimens were measured using real-time polymerase chain reaction. The clinical score of ocular surface findings was quantified using a 5-5-5 exacerbation grading scale for allergic conjunctivitis. The first case was of a 27-year-old man who developed DIOSD 3 months after starting treatment with dupilumab injection for atopic dermatitis. After 5 weeks of topical instillation of tacrolimus ophthalmic suspension, the clinical score of ocular surface findings improved and IL-8 and eotaxin-2 mRNA expression levels gradually decreased. The second patient was a 55-year-old man who developed DIOSD 11 weeks after the start of treatment with dupilumab injection for atopic dermatitis. Four weeks after starting ophthalmological treatment with tacrolimus ophthalmic suspension, his clinical scores on ocular surface findings improved and IL-8 mRNA expression levels decreased. The ocular surface test in this case revealed increased expression levels of IL-8 mRNA on the ocular surface at the onset of DIOSD, which decreased with the improvement of objective findings., Conclusions: DIOSD, which has been successfully treated with tacrolimus ophthalmic suspension, may involve IL-8-related inflammation in addition to type 2 inflammation., (© 2022. The Author(s).)

Purpose: We investigated ocular surface microbiota dysbiosis in patients with refractory allergic conjunctival diseases (ACDs; stratified into mild and severe groups) treated with topical tacrolimus., Methods: Patients (n = 21) with refractory ACDs (including vernal and atopic keratoconjunctivitis) actively treated with topical tacrolimus and 6 healthy controls were evaluated. Based on clinical scores and expression of specific cytokines on the ocular surface, patients with ACDs were divided into mild and severe groups using cluster analysis. The microbial composition of tear specimens collected from patients with mild and severe ACD and control subjects using the Schirmer test paper was determined through next-generation 16S rRNA sequencing analysis., Results: Compared with healthy controls, patients with ACDs exhibited significantly decreased ocular surface microbiota α-diversity. Ocular surface microbiota mainly comprised members of the phyla Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria in all groups. The relative abundance of ocular surface microbiota in patients with ACDs was increased for phylum Firmicutes and decreased for phylum Proteobacteria (compared with control subjects). The genera Blautia (vs. mild ACD group) and Morganella (vs. control group) exhibited significantly increased abundance only in the severe ACD group., Conclusions: The ocular surface microbiota in patients with severe ACD exhibited decreased diversity and exacerbation of dysbiosis compared with that in patients with mild ACD and control subjects. Patients with mild refractory ACD also exhibited decreased diversity of these microbiota. These alterations in microbiota indicated a change in the ocular surface of patients with refractory ACD (be it because of disease pathogenesis or topical immunomodulatory treatment)., Competing Interests: J. Shoji received honoraria from Santen Pharmaceutical Co., Ltd. and Senju Pharmaceutical Co., Ltd. G. Harata and F. He are employees of Takanashi Milk Products Co., Ltd. The remaining authors have no conflicts of interest to disclose., (Copyright © 2021 The Author(s). Published by Wolters Kluwer Health, Inc.)

Actinic keratosis (AK) is one of the most frequent pathologies in clinical dermatology. Approximately 10% of the prevalence of AK occurs at 30 years of age and rises to 80% at 70 years of life. This study conducted a split-face analysis based on Photodynamic Therapy (MALA-PDT) and 5-fluorouracil (5-FU) treatments in patients with AK. Clinical and histopathological studies evaluated each treatment's efficiency and side effects. Six patients were recruited between 50 and 70 years of age for this. MALA-PDT was applied on the right side of the face. A biodistribution time of 2 h, a dose of light of 36 J/cm 2 (20 min irradiation), a spot for photoirradiation with a total area of 100 cm 2 , and a wavelength of 635 nm were used. (LINCE equipment MMOPTICS-Brazil). By fluorescence diagnosis, clinical and subclinical AK lesions were evidenced in the area called the "cancerization field." Two cycles of MALA-PDT were performed on each patient, and 5-FU was applied twice daily for 15 days. A 2-month follow-up post-MALA-PDT and post-5-FU was carried out. The treatment based on 5-Flu and MALA-PDT clinically shows the same response, but MALA-PDT is less aggressive than 5-FU. The treatment of AK based on MALA-PDT is a practical, inexpensive, and easy-to-use technique., (Copyright © 2022. Published by Elsevier B.V.)

Although intranasal oxytocin is expected to be a novel therapy for the core symptoms of autism spectrum disorder, which has currently no approved medication, the efficacy of repeated administrations was inconsistent, suggesting that the optimal dose for a single administration of oxytocin is not optimal for repeated administration. The current double-blind, placebo-controlled, multicentre, crossover trial (ClinicalTrials.gov Identifier: NCT03466671) was aimed to test the effect of TTA-121, a new formulation of intranasal oxytocin spray with an enhanced bioavailability (3.6 times higher than Syntocinon® spray, as assessed by area under the concentration-time curve in rabbit brains), which enabled us to test a wide range of multiple doses, on autism spectrum disorder core symptoms and to determine the dose-response relationship. Four-week administrations of TTA-121, at low dose once per day (3 U/day), low dose twice per day (6 U/day), high dose once per day (10 U/day), or high dose twice per day (20 U/day), and 4-week placebo were administered in a crossover manner. The primary outcome was the mean difference in the reciprocity score (range: 0-14, higher values represent worse outcomes) on the Autism Diagnostic Observation Schedule between the baseline and end point of each administration period. This trial with two administration periods and eight groups was conducted at seven university hospitals in Japan, enrolling adult males with high-functioning autism spectrum disorder. Enrolment began from June 2018 and ended December 2019. Follow-up ended March 2020. Of 109 males with high-functioning autism spectrum disorder who were randomized, 103 completed the trial. The smallest P-value, judged as the dose-response relationship, was the contrast with the peak at TTA-121 6 U/day, with inverted U-shape for both the full analysis set (P = 0.182) and per protocol set (P = 0.073). The Autism Diagnostic Observation Schedule reciprocity score, the primary outcome, was reduced in the TTA-121 6 U/day administration period compared with the placebo (full analysis set: P = 0.118, mean difference = -0.5; 95% CI: -1.1 to 0.1; per protocol set: P = 0.012, mean difference = -0.8; 95% CI: -1.3 to -0.2). The per protocol set was the analysis target population, consisting of all full analysis set participants except those who deviated from the protocol. Most dropouts from the full analysis set to the per protocol set occurred because of poor adherence to the test drug (9 of 12 in the first period and 8 of 15 in the second period). None of the secondary clinical and behavioural outcomes were significantly improved with the TTA-121 compared with the placebo in the full analysis set. A novel intranasal spray of oxytocin with enhanced bioavailability enabled us to test a wide range of multiple doses, revealing an inverted U-shape dose-response curve, with the peak at a dose that was lower than expected from previous studies. The efficacy of TTA-121 shown in the current exploratory study should be verified in a future large-scale, parallel-group trial., (© The Author(s) (2022). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: Photodynamic therapy (PDT) activates a photosensitizer by visible light to generate cytotoxic oxygen species that lead to cell death. With proper illumination, PDT is often used in applications on superficial and sub-surface lesions. Sporotrichosis infection occurs by Sporothrix fungi which causes a skin wound, worsened by Candida albicans infections. This study investigated the photosensitizing efficiency of the Ru(phen) 2 (pPDIp)(PF 6 ) 2 complex, RupPDIp, against S. brasiliensis and C. albicans., Material and Methods: RupPDIp efficiency against these fungi was tested using 450 nm (blue light and 36 J/cm 2 ) and 525 nm (green light, 25.2 J/cm 2 ) at 0.05-20 μM concentrations. To ensure PDT effectiveness, control groups were tested in the absence and in the presence of RupPDIp under light irradiation and in the dark., Results: RupPDIp eliminated both fungi at ≤5.0 μM. Green light showed the best results, eliminating S. brasiliensis and C. albicans colonies at RupPDIp 0.5 μM and 0.05 μM, respectively., Conclusion: RupPDIp is a promising photosensitizer in aPDT, eliminating 10 6  CFU/mL of both fungi at 450 nm and 525 nm, with lower light doses and concentrations when treated with the green light compared to the blue light., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: To date, the usefulness of parathyroid hormone [PTH (1-34)] in distraction osteogenesis has been reported in several studies. We aimed to determine the optimal timing of PTH (1-34) administration in a rabbit distraction osteogenesis model., Methods: The lower hind leg of a Japanese white rabbit was externally fixed, and tibial osteotomy was performed. One week after the osteotomy, bone lengthening was carried out at 0.375 mm/12 h for 2 weeks. After 5 weeks, the lower leg bone was collected. Bone mineral density (BMD), peripheral quantitative computed tomography (pQCT), micro-computed tomography (micro-CT), and mechanical tests were performed on the distracted callus. The rabbits were divided into three groups according to the timing of PTH (1-34) administration: 4 weeks during the distraction and consolidation phases (group D + C), 2 weeks of the distraction phase (group D), and the first 2 weeks of the consolidation phase (group C). A control group (group N) was administered saline for 4 weeks during the distraction and consolidation phases. Furthermore, to obtain histological findings, lower leg bones were collected from each rabbit at 2, 3, and 4 weeks after osteotomy, and tissue sections of the distracted callus were examined histologically., Results: The BMD was highest in group C and was significantly higher than group D. In pQCT, the total cross-sectional area was significantly higher in groups D + C, D, and C than group N, and the cortical bone area was highest in group C and was significantly higher than group D. In micro-CT, group C had the highest bone mass and number of trabeculae. Regarding the mechanical test, group C had the highest callus failure strength, and this value was significantly higher compared to group N. There was no significant difference between groups D and N. The histological findings revealed that the distracted callus mainly consisted of endochondral ossification in the distraction phase. In the consolidation phase, the chondrocytes were almost absent, and intramembranous ossification was the main type of ossification., Conclusion: We found that the optimal timing of PTH (1-34) administration is during the consolidation phase, which is mainly characterized by intramembranous ossification., (© 2022. The Author(s).)

Human Papillomavirus (HPV) infection is highly prevalent worldwide, and one of its consequences is the external genital wart, or Condyloma Acuminata (CA). The present study used Photodynamic Therapy (PDT) to treat CA lesions. PDT treated 23 patients with a clinical diagnosis of multifocal and unifocal CA. Patients were divided into Group 1 (G1, Patients without pathologies associated with immunodeficiency) and Group 2 (G2, patients with pathologies associated with immunodeficiency). In the G1 group (19 patients), PDT resulted in a Complete Response in 68.4% (average 5 PDT cycles), Partial Response in 26.3% (average 10 PDT cycles), and No Response in 5.3% (average 6 PDT cycles). In the G2 group (4 patients), 100% of subjects showed a partial response (8 PDT cycles). These patients in the G2 and with partial response had associated pathologies, such as renal failure, breast cancer, and HIV. There was a slight decrease in lesions (20-40%) post-treatment in these cases. Four months after treatment, no new lesions or recurrence were observed in the entire area treated with PDT using low doses of PDT. Eighty-six percent of the patients tolerated the treatments well. We conclude that PDT is a promising and safe treatment for CA lesions compared to traditional treatments., (Copyright © 2022. Published by Elsevier B.V.)

Objective: Time-processing disorders in adults is a priority area for intervention. Time management program, which has been demonstrated to be effective in children with ADHD, has not been examined in adults. We anticipate the need for the development of specialized programs for adults. This is because it has been reported that time processing disorders have different patterns in childhood and adulthood. This study aimed to evaluate the therapeutic effect of a gCBT program focusing on time management for adults with ADHD., Method: Adults with ADHD were randomly assigned to gCBT ( n  = 24) or a treatment as usual group ( n  = 24). Outcome measures were masked clinically rated, self-reported, and family-reported ADHD symptoms., Results: The gCBT group significantly reduced ADHD symptoms on all measures., Conclusion: Interventions focused on time management have been shown to be effective not only in children with ADHD but also in adult patients.

Purpose: To evaluate the clinical improvement and safety of prolonged treatment of vernal (VKC) and atopic keratoconjunctivitis (AKC) using topical tacrolimus., Methods: We included 36 eyes of 36 patients who had VKC and AKC and were treated with topical tacrolimus ophthalmic suspension (0.1%) for 24 months. The demographic data of the enrolled patients were collected from their medical files. Clinical scores, remission rates, number of relapses, concomitant use of steroids, and refractory indices were assessed. Clinical outcomes were determined using papillae-limbus-cornea (PLC) scores and 5-5-5 exacerbation grading scale scores. Clinical characteristics associated with the need for concomitant steroid eye drops administration were determined using logistic regression analysis. All patients were classified into 3 subgroups using cluster analysis., Results: PLC scores recorded in the sixth month were significantly improved compared with those recorded at baseline. PLC scores recorded in the 18th, 21st, and 24th months were significantly improved compared with those recorded in the sixth month. The remission rates increased diachronically and significantly, reaching 92% in the 24th month. Logistic regression analysis showed that, for every 10-year increase in patient age, the risk for requiring concomitant administration of steroid eye drops was reduced by half (odds ratio, 0.53; 95% confidence interval, 0.29-0.96). Using cluster analysis, the patients were divided into 3 clusters: adolescent type, pediatric type, and adult type., Conclusions: Two years of treatment with topical tacrolimus ophthalmic suspension is an effective method for inducing and maintaining the stable stages of VKC and AKC., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2021 The Author(s). Published by Wolters Kluwer Health, Inc.)

Azide-oligo(ethylene glycol)-terminated self-assembled monolayers (N 3 -OEG-SAMs) are promising interfacial structures for surface functionalization. Its many potential applications include chemical/bio-sensing and construction of surface models owing to its cross-coupling activity that originates from the azide group and oligo(ethylene glycol) (OEG) units for non-specific adsorption resistance. However, there are only a few studies and limited information, particularly on the molecular-scale structures and local cross-coupling activities of N 3 -OEG-SAMs, which are vital to understanding its surface properties and interfacial molecular design. In this study, molecular-scale surface structures and cross-coupling activity of azide-hexa(ethylene glycol)-terminated SAMs (N 3 -EG 6 -SAMs) were investigated using frequency modulation atomic force microscopy (FM-AFM) in liquid. The N 3 -EG 6 -SAMs were prepared on Au(111) substrates through the self-assembly of 11-azido-hexa(ethylene glycol)-undecane-1-thiol (N 3 -EG 6 -C 11 -HS) molecules obtained from a liquid phase. Subnanometer-resolution surface structures were visualized in an aqueous solution using a laboratory-built FM-AFM instrument. The results show a well-ordered molecular arrangement in the N 3 -EG 6 -SAM and its clean surfaces originating from the adsorption resistance property of the terminal EG 6 units. Surface functionalization by the cross-coupling reaction of copper(I)-catalyzed azide-alkyne cycloaddition was observed, indicating a structural change in the form of fluctuating structures and island-shaped structures depending on the concentration of the alkyne molecules. The FM-AFM imaging enabled to provide information on the relationship between the surface structures and cross-coupling activity. These findings provide molecular-scale information on the functionalization of the N 3 -EG 6 -SAMs, which is helpful for the interfacial molecular design based on alkanethiol SAMs in many applications.

Endoreplication is a type of cell cycle where genome replication occurs without mitosis. An increase of ploidy level by endoreplication is often associated with cell enlargement and an enhanced plant growth. Here we report Arabidopsis thaliana subclass I ACTIN DEPOLYMERIZING FACTORs (ADFs) and vegetative ACTIN2/8 as novel regulators of endoreplication. A. thaliana has 11 ADF members that are divided into 4 subclasses. Subclass I consists of four members, ADF1, -2, -3, and -4, all of which constitutively express in various tissues. We found that both adf4 knockout mutant and transgenic plants in which expressions of all of four subclass I ADFs are suppressed (ADF1-4Ri) showed an increased leaf area of mature first leaves, which was associated with a significant increase of epidermal pavement cell area. Ploidy analysis revealed that the ploidy level was significantly increased in mature leaves of ADF1-4Ri. The increased ploidy was also observed in roots of adf4 and ADF1-4Ri, as well as in dark-grown hypocotyls of adf4. Furthermore, double mutants of vegetative ACT2 and ACT8 (act2/8) exhibited an increase of leaf area and ploidy level in mature leaves. Therefore, actin-relating pathway could regulate endoreplication. The possible mechanisms that actin and ADFs regulate endoreplication are discussed., (© 2021. The Botanical Society of Japan.)

A simple methylenedioxy dibromoindole alkaloid, amakusamine ( 1 ), was isolated from a marine sponge of the genus Psammocinia , and its structure was determined from spectroscopic data, time-dependent density-functional theory calculations, and synthesis. Compound 1 inhibited the receptor activator of nuclear factor-κB ligand (RANKL)-induced formation of multinuclear osteoclasts with an IC 50 value of 10.5 μM in RAW264 cells. The structure - activity relationship of 1 was also investigated with synthetic derivatives.

Background and Objectives: This work evaluated antimicrobial photodynamic therapy (PDT), sonodynamic therapy (SDT), and the association of both therapies (sonophotodynamic therapy [SPDT]), mediated by curcumin (Cur) against Staphylococcus aureus biofilm. Next, additional strategies for these treatments were assessed., Materials and Methods: S. aureus biofilms received PDT, SDT, and SPDT, mediated by Cur (80 µM), LED light (450 nm), and 1 MHz ultrasound. The same treatments were also performed adding a strategy: Cur with sodium dodecyl sulfate (SDS), Cur with potassium iodide (KI) or a pre-treatment with ultrasound. Cell viability was determined and biofilm architecture was evaluated under confocal microscopy., Results: SPDT was more effective to inactivate the bacteria than PDT and SDT. SDS achieved the greatest viability reductions, followed by KI and ultrasound pre-treatment. Confocal images revealed biofilm disruption and a reduced number of cells in all treatments. However, SPDT exhibited a pronounced effect and it was greater using SDS., Conclusion: SPDT was more effective and additional strategies potentiated its effectiveness. Lasers Surg. Med. © 2021 Wiley Periodicals LLC., (© 2021 Wiley Periodicals LLC.)

Fifteen new isopimarane-type diterpenes, taichunins E-S ( 1 - 15 ), and a new 20- nor -isopimarane, taichunin T ( 16 ), together with four known compounds were isolated from Aspergillus taichungensis (IBT 19404). The structures of these new compounds were determined by NMR and mass spectroscopy, and their absolute configurations were analyzed by NOESY and TDDFT calculations of ECD spectra. Taichunins G, K, and N ( 3 , 7 , and 10 ) completely inhibited the receptor activator of nuclear factor-κB ligand (RANKL)-induced formation of multinuclear osteoclasts in RAW264 cells at 5 μM, with 3 showing 92% inhibition at a concentration of 0.2 μM.

Purpose: To investigate whether crude house-dust-mite antigen exacerbates eosinophilic inflammation in the conjunctival tissues of an atopic keratoconjunctivitis mouse model in a dose-dependent manner., Materials and Methods: An atopic keratoconjunctivitis mouse model was established by percutaneous sensitization and crude house-dust-mite antigen application in NC/Nga mice. To assess the dose-dependent response, conjunctival specimens from groups that were administered high- (High-HDM) or low-dose house-dust-mite antigen (Low-HDM) following percutaneous sensitization and the control without house-dust-mite antigen administration (control group) were evaluated. Histological examination and immunofluorescence staining were performed to determine eosinophil density and the number of IL-13-positive cells. Polymerase chain reaction array was used to obtain adaptive and innate immunity-related factor profile, and quantitative polymerase chain reaction was used to determine Il13, Il17a, Ccl11, and Ccl24 expression. Atopic keratoconjunctivitis model mice injected with anti-IL-1α antibody (IL-1α group) or vehicle (vehicle group) to the upper and lower eyelids before atopic keratoconjunctivitis development were evaluated., Results: Eosinophil density in the conjunctiva increased with house-dust-mite antigen application in a dose-dependent manner. CD4, CXCL10, CCR6, C3, and IL-13 mRNA levels increased more than 5-fold in the conjunctiva of the High-HDM group animals compared to those in control animals. mRNA expression of Il13 and Ccl11 in the conjunctiva of the High-HDM group animals significantly increased compared with that in the Low-HDM and control group animals. Conversely, the eosinophil density and Il13 mRNA expression significantly decreased in the IL-1α group compared with those in the vehicle group., Conclusions: The house-dust-mite antigen increased eosinophilic infiltration and Il13 mRNA expression in the conjunctiva of an atopic keratoconjunctivitis mouse model in a dose-dependent manner. These inflammatory alterations were partially alleviated by eyelid injection of anti-IL-1α antibody. These findings indicate that IL-1α-induced IL-13 production constitutes a major exacerbating factor for house-dust-mite antigen-induced atopic keratoconjunctivitis.

Background: The Behavior Problems Inventory-Short Form (BPI-S), which assesses behavior problems in individuals with intellectual disabilities (ID), is a shorter version of the BPI-01. This study investigated the reliability and validity of the BPI-S Japanese version (BPI-S-J) for adolescents/adults with ID and behavior problems., Methods: The test-retest reliability included participants with ID and behavioral problems who were enrolled in welfare services. For test-retest reliability, 42 caregivers independently responded to the BPI-S-J every two weeks. Inter-rater reliability was independently assessed using the BPI-S-J by two caregivers who were familiar with the 42 participants. The participants of the validity assessment were 227 students from special needs schools or patients with ID admitted to medical institutions. The total frequency total score was compared based on the degree of ID. To examine the criterion-related validity, we analyzed the total frequency score, the total score of the Criteria for Determining Severe Problem Behavior (CDSPB) and the total score of the Aberrant Behavior Checklist-Japanese version (ABC-J)., Results: The BPI-S-J of test-retest reliability was satisfactory (intra-class correlation; ICC) = 0.954), and the total score significantly (ICC = 0.721) represented good inter-rater reliability. For the validity, the BPI-S-J score of participants who had severe and profound ID was significantly higher than those who had mild and moderate ID. Significant correlations were observed between the BPI-S-J score and CDSPB score (r = 0.499), and the ABC-J score (r = 0.699), indicating adequate criterion-related validity., Conclusion: This study showed the utility of the BPI-S-J to assess behavior problems in the Japanese ID population., (Copyright © 2021 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.)

Purpose: To evaluate the presence of ocular surface mucin in patients with atopic and vernal keratoconjunctivitis (AKC/VKC), we investigated the mRNA expression levels of SAM-pointed domain-containing ETS-like factor ( SPDEF ) and mucin-related genes on the ocular surface., Methods: Nineteen patients with AKC or VKC were divided into two groups based on the severity of the disease as determined by their clinical scores for AKC/VKC: the stable group and the active group. Impression cytology was performed in all patients using filter paper, and the expression levels of SPDEF , MUC1 , MUC4 , MUC5AC , MUC16 , and eotaxin-2 mRNA were determined by real-time reverse-transcription polymerase chain reaction., Results: The results showed that the expression levels of SPDEF and MUC5AC mRNA in the active group were significantly decreased compared with those in the stable group. Furthermore, clinical scores were significantly negatively correlated with the expression levels of SPDEF mRNA and significantly positively correlated with the expression levels of eotaxin-2, which is a biomarker for eosinophilic inflammation on the ocular surface. Cluster analysis classified the patients with AKC/VKC into three clusters, and the stable group was divided into two clusters according to the condition of ocular surface mucin., Conclusions: Ocular surface mucin in patients with AKC/VKC is altered in accordance with the clinical severity of the disease., Competing Interests: J. S. has previously received honoraria from Santen Pharmaceutical Co., Ltd., Senju Pharmaceutical Co., Ltd., and Alcon Pharmaceuticals, outside the submitted work. S. Y., N. I., Y. T., A. T., and M. H. declare that they have no conflicts of interest., (Copyright © 2021 Mariko Horinaka et al.)

Glycosylinositolphosphorylceramides (GIPCs) are the predominant lipid in the outer leaflet of the plasma membrane. Characterized GIPC glycosylation mutants have severe or lethal plant phenotypes. However, the function of the glycosylation is unclear. Previously, we characterized Arabidopsis thaliana GONST1 and showed that it was a nucleotide sugar transporter which provides GDP-mannose for GIPC glycosylation. gonst1 has a severe growth phenotype, as well as a constitutive defense response. Here, we characterize a mutant in GONST1's closest homolog, GONST2. The gonst2- 1 allele has a minor change to GIPC headgroup glycosylation. Like other reported GIPC glycosylation mutants, gonst1-1gonst2-1 has reduced cellulose, a cell wall polymer that is synthesized at the plasma membrane. The gonst2-1 allele has increased resistance to a biotrophic pathogen Golovinomyces orontii but not the necrotrophic pathogen Botrytis cinerea . Expression of GONST2 under the GONST1 promoter can rescue the gonst1 phenotype, indicating that GONST2 has a similar function to GONST1 in providing GDP-D-Man for GIPC mannosylation., Competing Interests: No conflicts of interest to declare., (© 2021 The Authors. Plant Direct published by American Society of Plant Biologists, Society for Experimental Biology and John Wiley & Sons Ltd.)

Objective: Brown adipose tissue (BAT) is specialized in thermogenesis. The conversion of energy into heat in brown adipocytes proceeds via stimulation of β-adrenergic receptor (βAR)-dependent signaling and activation of mitochondrial uncoupling protein 1 (UCP1). We have previously demonstrated a functional role for pannexin-1 (Panx1) channels in white adipose tissue; however, it is not known whether Panx1 channels play a role in the regulation of brown adipocyte function. Here, we tested the hypothesis that Panx1 channels are involved in brown adipocyte activation and thermogenesis., Methods: In an immortalized brown pre-adipocytes cell line, Panx1 currents were measured using patch-clamp electrophysiology. Flow cytometry was used for assessment of dye uptake and luminescence assays for adenosine triphosphate (ATP) release, and cellular temperature measurement was performed using a ratiometric fluorescence thermometer. We used RNA interference and expression plasmids to manipulate expression of wild-type and mutant Panx1. We used previously described adipocyte-specific Panx1 knockout mice (Panx1 Adip-/- ) and generated brown adipocyte-specific Panx1 knockout mice (Panx1 BAT-/- ) to study pharmacological or cold-induced thermogenesis. Glucose uptake into brown adipose tissue was quantified by positron emission tomography (PET) analysis of 18 F-fluorodeoxyglucose ( 18 F-FDG) content. BAT temperature was measured using an implantable telemetric temperature probe., Results: In brown adipocytes, Panx1 channel activity was induced either by apoptosis-dependent caspase activation or by β3AR stimulation via a novel mechanism that involves Gβγ subunit binding to Panx1. Inactivation of Panx1 channels in cultured brown adipocytes resulted in inhibition of β3AR-induced lipolysis, UCP-1 expression, and cellular thermogenesis. In mice, adiponectin-Cre-dependent genetic deletion of Panx1 in all adipose tissue depots resulted in defective β3AR agonist- or cold-induced thermogenesis in BAT and suppressed beigeing of white adipose tissue. UCP1-Cre-dependent Panx1 deletion specifically in brown adipocytes reduced the capacity for adaptive thermogenesis without affecting beigeing of white adipose tissue and aggravated diet-induced obesity and insulin resistance., Conclusions: These data demonstrate that Gβγ-dependent Panx1 channel activation is involved in β3AR-induced thermogenic regulation in brown adipocytes. Identification of Panx1 channels in BAT as novel thermo-regulatory elements downstream of β3AR activation may have therapeutic implications., (Copyright © 2020 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Ocular surface mucins are thought to play vital roles in maintaining the homeostasis of the pre-ocular surface tear film. We performed ocular surface tests with impression cytology to assess the expression levels of mucin-related genes on the ocular surface in healthy eyes. In addition, we investigated alterations in mucin-related gene expression secondary to treatment with rebamipide ophthalmic suspension in patients with Sjögren's syndrome-associated dry eyes (SS-DE). Thirty-three healthy individuals (control group) and 13 patients from our hospital with SS-DE were enrolled. Impression cytology was performed using Schirmer's test paper for RNA sampling. The mRNA levels of SAM-pointed domain-containing ETS-like factor (SPDEF), mucin 5AC (MUC5AC), and mucin 16 (MUC16) were determined using a real-time reverse transcription-polymerase chain reaction. The ocular surface test was performed once for the control group, and at baseline as well as 2, 4, 8, and 12 weeks after treatment in the Sjögren's syndrome-associated dry eyes group. mRNA levels of SPDEF, MUC5AC, and MUC16 were not significantly different between the control and SS-DE groups before rebamipide ophthalmic suspension treatment. SPDEF mRNA levels in control subjects were significantly correlated with levels of MUC5AC. Among SS-DE patients, SPDEF mRNA levels were significantly increased at 2, 4, and 8 weeks after treatment compared with baseline levels. MUC16 mRNA levels were significantly decreased from baseline levels at 4 and 8 weeks post-treatment. Ocular surface test using impression cytology is a clinically useful tool for assessing mucous conditions on the ocular surface and can be used to determine the effects of instillation treatment with eye drops that affect mucin production at the ocular surface., Competing Interests: Financial support for this research was received from the Otsuka Pharmaceutical Co., Ltd., Japan. I have read the journal's policy and the authors of this manuscript have the following competing interests: J.S. received personal fees from Santen Pharmaceutical Co. Ltd., Senju Pharmaceutical Co. Ltd., Alcon Pharmaeuticals, outside the submitted work. Y.S., N.I., and A.T. declare that they have no conflict of interest. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Background: One aim of an autism spectrum disorder (ASD) diagnosis is to obtain special support for the disorder, though this does not guarantee practical support. We developed a psychoeducational program using cognitive-behavioral therapy (CBT) and Aware and Care for my Autistic Traits (ACAT) for Japanese adolescents with high-functioning ASD and their parents., Methods: This multisite study is a randomized controlled trial. In total, 24 participants will be assigned to the ACAT group and 24 to the treatment-as-usual (TAU) group. The ACAT group will receive a weekly 100-min session for 6 weeks, regular medical care, and one follow-up session. In this ongoing clinical trial, we will compare the scores of the measures recorded in the pre- and post-intervention stages between the ACAT and TAU groups. A total of 41 patients out of a target of 48 have participated in the trial to date. The primary outcome measure is the Autism Knowledge Questionnaire. Secondary outcome measures include Barriers to Access to Care Evaluation 3rd Edition, the Strengths and Difficulties Questionnaire, the Vineland Adaptive Behavior Scales second edition, the Parenting Resilience Elements Questionnaire, the General Health Questionnaire 12, and the Depression Self-Rating Scale for Children assessments, as well as an electroencephalographic recording., Discussion: It is expected that participants in the ACAT group will significantly increase their self-understanding and awareness of ASD symptoms compared to those in the TAU group. Additionally, the ACAT group is expected to exhibit improved social adaptation and mental health if children and parents are able to better understand the ASD characteristics through sessions. This intervention will contribute to the establishment of an effective evidence-based treatment strategy for adolescents with ASD., Trial Registration: UMIN Register 000029851 . Registered on January 06, 2018.

Purpose: To report an atypical presentation of herpes simplex virus (HSV) keratitis followed up using expression levels of HSV DNA in tears., Methods: A 22-year-old Japanese woman with hyperemia and foreign body sensation in her left eye was diagnosed with atypical dendritic keratitis. A slit-lamp examination at presentation indicated the presence of a rush of dendritic lesions with a sparse branching pattern and poor development of terminal bulbs; follicular conjunctivitis was also observed. Positivity for house-dust-mite- and cedar pollen-specific IgE antibodies in her serum indicated atopic diathesis. The HSV DNA levels in her tears were measured by a real-time polymerase chain reaction., Results: At the initial visit, the HSV DNA levels in tears were 6.4 × 10 copies/sample in the right eye and 1.6 × 10 copies/sample in the left eye. The keratitis improved after treatment with topical acyclovir ointment, 5 times a day for 7 days, and systemic valacyclovir 1000 mg/d for 5 days. Multiple punctate subepithelial opacities developed in her left eye on day 7, with undetectable HSV DNA in tears, bilaterally., Conclusions: We have successfully monitored the HSV DNA levels in tears using quantitative real-time polymerase chain reaction in HSV keratitis where the corneal findings progressed from atypical dendritic keratitis to multiple punctate corneal subepithelial opacities during the treatment period.

Chromosome instability (CIN), the hallmarks of cancer, reflects ongoing chromosomal changes caused by chromosome segregation errors and results in whole chromosomal or segmental aneuploidy. In multiple myeloma (MM), CIN contributes to the acquisition of tumor heterogeneity, and thereby, to disease progression, drug resistance, and eventual treatment failure; however, the underlying mechanism of CIN in MM remains unclear. Faithful chromosomal segregation is tightly regulated by a series of mitotic checkpoint proteins, such as budding uninhibited by benzimidazoles 1 (BUB1). In this study, we found that BUB1 was overexpressed in patient-derived myeloma cells, and BUB1 expression was significantly higher in patients in an advanced stage compared to those in an early stage. This suggested the involvement of aberrant BUB1 overexpression in disease progression. In human myeloma-derived cell lines (HMCLs), BUB1 knockdown reduced the frequency of chromosome segregation errors in mitotic cells. In line with this, partial knockdown of BUB1 showed reduced variations in chromosome number compared to parent cells in HMCLs. Finally, BUB1 overexpression was found to promote the clonogenic potency of HMCLs. Collectively, these results suggested that enhanced BUB1 expression caused an increase in mitotic segregation errors and the resultant emergence of subclones with altered chromosome numbers and, thus, was involved in CIN in MM.

Photonic crystals are some of the more spectacular realizations that periodic arrays can change the behavior of electromagnetic waves. In nature, so-called structural colors appear in insects and even plants. Some species create beautiful color patterns as part of biological behavior such as reproduction or defense mechanisms as a form of biomimetics. The interaction between light and matter occurs at the surface, producing diffraction, interference and reflectance, and light transmission is possible under suitable conditions. In particular, there are two Colombian butterflies, Morpho cypris and Greta oto, that exhibit iridescence phenomena on their wings, and in this work, we relate these phenomena to the photonic effect. The experimental and theoretical approaches of the optical response visible region were studied to understand the underlying mechanism behind the light-matter interaction on the wings of these Colombian butterflies. Our results can guide the design of novel devices that use iridescence as angular filters or even for cosmetic purposes.

Background: The dispersal of airborne norovirus (NoV) particles from the floor after contamination with faeces or vomit is a challenge for infection control, as this pathogen is infectious at low doses. Therefore, it is imperative to establish a safe protocol for floor decontamination., Aim: To assess the presence of residual NoV-GII particles on floors and airborne particles following various floor decontamination procedures., Methods: Two types of floor (vinyl and granite) were contaminated intentionally with 10% human faeces, positive for NoV-GII. Two decontamination protocols were implemented: cleaning followed by disinfection using 1% sodium hypochlorite, and cleaning followed by disinfection using a manual ultraviolet C (UV-C) light device. Swab samples were taken from the floors, and air samples were obtained using an air sampler. The TaqMan method for real-time reverse transcription-quantitative polymerase chain reaction was employed for analysis., Findings: The disinfection protocol using 1% sodium hypochlorite after cleaning proved to be more effective than cleaning followed by UV-C light exposure (P<0.001). Viral particles were detected in 27 of 36 air samples after cleaning, with no significant difference between the two floor types. On average, 617 genome copies/sample were identified in air samples after cleaning, but the number decreased gradually after disinfection., Conclusion: NoV-GII can be aerosolized during floor cleaning, and its particles may be inhaled and then swallowed or can settle on surfaces. Therefore, residual viral particles on floors must be fully eliminated. Cleaning followed by 10 min of 1% sodium hypochlorite disinfection proved to be the superior decontamination protocol., (Copyright © 2019 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

Decreases in photosynthetic rate, stomatal conductance (gs), and mesophyll conductance (gm) are often observed under elevated CO2 conditions. However, which anatomical and/or physiological factors contribute to the decrease in gm is not fully understood. Arabidopsis thaliana wild-type and carbon-metabolism mutants (gwd1, pgm1, and cfbp1) with different accumulation patterns of non-structural carbohydrates were grown at ambient (400 ppm) and elevated (800 ppm) CO2. Anatomical and physiological traits of leaves were measured to investigate factors causing the changes in gm and in the mesophyll resistance (expressed as the reciprocal of mesophyll conductance per unit chloroplast surface area facing to intercellular space, Sc/gm). When grown at elevated CO2, all the lines showed increases in cell wall mass, cell wall thickness, and starch content, but not in leaf thickness. gm measured at 800 ppm CO2 was significantly lower than at 400 ppm CO2 in all the lines. Changes in Sc/gm were associated with thicker cell walls rather than with excess starch content. The results indicate that the changes in gm and Sc/gm that occur in response to elevated CO2 are independent of non-structural carbohydrates, and the cell wall represents a greater limitation factor for gm than starch., (© The Author(s) 2019. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

Cationic nanogels of N -isopropylacrylamide (NIPAM), including NIPAM-based cationic fluorescent nanogel thermometers, were synthesized with a cationic radical initiator previously developed in our laboratory. These cationic nanogels were characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), zeta potential measurements and fluorescence spectroscopy, as summarized in the temperature-dependent fluorescence response based on the structural change in polyNIPAM units in aqueous solutions. Cellular experiments using HeLa (human epithelial carcinoma) cells demonstrated that NIPAM-based cationic fluorescent nanogel thermometers can spontaneously enter the cells under mild conditions (at 25 °C for 20 min) and can show significant fluorescence enhancement without cytotoxicity with increasing culture medium temperature. The combination of the ability to enter cells and non-cytotoxicity is the most important advantage of cationic fluorescent nanogel thermometers compared with other types of fluorescent polymeric thermometers, i.e., anionic nanogel thermometers and cationic/anionic linear polymeric thermometers.

Pathogenic fungi from the genus Colletotrichum form invasive hyphae; the hyphae are surrounded by an extra-invasive hyphal membrane (EIHM), which is continuous with the plant plasma membrane. Although the EIHM plays a crucial role as the interface between plant and fungal cells, its precise function during Colletotrichum infection remains elusive. Here, we show that enrichment of phosphoinositides (PIs) has a crucial role in Colletotrichum infection. We observed the localization of PIs in Arabidopsis thaliana cells infected by A. thaliana-adapted Colletotrichum higginsianum (Ch), and found that phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] was extremely enriched in the EIHM during Ch infection. We also found that phosphatidylinositol 4-phosphate-5 kinase (PIP5K), which catalyzes production of PI(4,5)P2, also accumulated at the EIHM. The overexpression of PIP5K3 in A. thaliana increased hyphal invasion by Ch. An exocytic factor, EXO84b, was targeted to the EIHM during Ch infection, although endocytic factors such as CLATHRIN LIGHT CHAIN 2 and FLOTILLIN 1 did not. Intriguingly, the interfacial membranes between A. thaliana and powdery mildew- or downy mildew-causing pathogens did not accumulate PI(4,5)P2. These results suggest that Ch could modify the PI(4,5)P2 levels in the EIHM to increase the exocytic membrane/protein supply of the EIHM for successful infection. Our results also suggest that PI(4,5)P2 biosynthesis is a promising target for improved defense against Colletotrichum infection., (� 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.)

Purpose: We validated the use of chemokine messenger RNA (mRNA) expression analysis for the assessment of ocular surface allergic inflammation in chronic allergic conjunctival diseases (ACDs) with proliferative lesions, including giant papillae and gelatinous infiltration of the limbus., Methods: This prospective sectional study included 19 patients with chronic ACDs and 10 healthy volunteers as controls. Patients with chronic ACDs were divided into 2 subgroups according to the severity of the clinical score: active stage ACD subgroup (n = 9) and stable stage ACD subgroup (n = 10). Impression cytology using a filter paper for each upper tarsal conjunctiva of the patients with chronic ACDs and control subjects was performed, and the expression levels of IL1A, CXCL8, IL16, and CCL24 mRNAs encoding interleukin (IL)-1α, CXCL8/IL-8, IL-16, and CCL24/eotaxin-2, respectively, were determined by quantitative real-time polymerase chain reaction using impression cytology specimens., Results: CCL24 and IL16 mRNA levels in the active ACD subgroup were significantly higher than those in the control group (P = 0.003 and 0.004, respectively). IL1A and CXCL8 expression levels in the active ACD subgroup were significantly higher than those in the stable ACD (P = 0.008 and 0.029, respectively) and control (P = 0.008 and 0.014, respectively) subgroups. Furthermore, significant correlations were detected between IL16 and CCL24 mRNA levels (r = 0.76, P = 0.0001) and between IL1A and CXCL8 (r = 0.67, P = 0.0004)., Conclusions: At least 2 kinds of inflammatory reactions, IL-1α- and CXCL8-associated inflammation and CCL24- and IL-16-associated inflammation, may be involved in the exacerbation of chronic ACDs.

This study is a retrospectively recruited case series. We report three infants with acute conjunctivitis induced by β-lactamase-positive, ampicillin/clavulanic acid-resistant strains of Haemophilus influenzae (BLPACR). Patients with BLPACR-positive cultures were recruited from among 5,107 patients with inflammatory diseases of the ocular surface who underwent examinations, including bacterial culturing of conjunctival sac or corneal scrapings, between 2000 and 2015. Three BLPACR-positive patients were recruited, including a 10-month-old boy, a 4-month-old girl, and a 7-month-old girl. All three demonstrated BLPACR conjunctivitis. The clinical findings in these patients included fever, mucopurulent discharge, lid swelling, and conjunctival hyperemia. Samples of conjunctival swabs were obtained from all three infants, and BLPACR was isolated from all these conjunctival swabs. Antimicrobial susceptibility testing showed sensitivity to levofloxacin and resistance to ampicillin, cefaclor, and clarithromycin. We conclude that in infantile BLPACR conjunctivitis, simultaneous investigation for the determination of causative organism and antibiotic susceptibility testing are crucial aspects of the medical treatment.

Temperature is one of the most important of the physiological parameters that determine the biological status of living organisms. However, intracellular temperature was not imaged at the single-cell level until recently because of the lack of a molecular thermometer that can be applied to living cells. We have recently developed a method for imaging intracellular temperature using a cationic linear fluorescent polymeric thermometer (FPT) and fluorescence lifetime imaging microscopy (FLIM). The cationic linear FPT exhibits cell permeability in various mammalian cell lines and yeast cells, entering live cells within 10 min of incubation. Intracellular thermometry using the cationic linear FPT and FLIM can be used to image temperature with high temperature resolution (0.3-1.29 °C within a temperature range of 25-35 °C). The diffuse intracellular localization of the cationic linear FPT allows a high spatial resolution (i.e., the light microscope's diffraction limit, 200 nm), enabling the detection of temperature distributions at the subcellular level. This protocol, including the construction of a calibration curve and intracellular temperature imaging, requires ~14 h. Experience in handling cultured mammalian cells and use of a confocal laser-scanning microscope (CLSM) is required.

Core-shell nanoparticles operating by infrared-to-visible energy upconversion (UCNPs) have been proposed as theranostic carriers for photosensitizers to increase deep-tissue penetration of photodynamic therapy against tumors and bacterial infections. Herein we present a series of core-shell mesoporous silica-coated NaYF 4 :Yb:Er UCNPs (mSiO 2 @UCNP) with different surface functionalizations to enhance bacterial targeting and loaded with the hydrophobic photosensitizer SiPc (silicon 2,9,16,23-tetra- tert -butyl-29 H ,31 H -phthalocyanine dihydroxide) to boost the bactericidal effect against Gram-positive and Gram-negative bacteria upon near-infrared irradiation. Förster resonance energy transfer (FRET) from the UCNP core to loaded SiPc was facilitated, while its efficiency depended on UCNP shell functionalization, which influences the SiPc penetration depth into the mesoporous silica, constituting a convenient tool to modify FRET intensity. Functionalized UCNPs displayed dark toxicity toward Gram-negative E. coli of up to 5 orders of magnitude, while Gram-positive S. aureus viability was not decreased in the dark, offering practical means for discriminating between the two bacterial strains. Directly exciting SiPc on the UNCP led to complete eradication of E. coli and a drastic decrease of colony-forming units of S. aureus of up to 7 orders of magnitude. With this study, we demonstrate strategies to potentiate antimicrobial photodynamic therapy on nanoparticular structures that can lead to next-generation photosensitizing systems based on UCNPs to help encounter and eradicate resistant bacteria, as well as for theranostics and future in vivo applications.

The primary objective of this study was to describe parameters related to the leaf biochemical assimilation capacity of photosynthesis, such as the maximum rates of carboxylation (V cmax ) and electron transport (J max ), as a function of the leaf nitrogen content throughout a canopy of Siebold's beech and Japanese oak grown under elevated ozone (O 3 ) conditions during a growing season. To this end, we investigated the relationship between photosynthetic traits and leaf nitrogen content in various canopy positions of two tree species under free-air O 3 exposure (60 nmol mol -1 , during daylight hours) in June, August, and October 2012. We observed O 3 -induced reduction in V cmax and J max without reduction of leaf nitrogen content in both tree species. In Siebold's beech, V cmax and J max in leaves with higher N area were largely decreased by O 3 from August, while little effect of O 3 was observed in leaves with lower N area . On the other hand, there was no difference in the extent of O 3 -induced reduction in V cmax and J max across the range of N area in leaves of Japanese oak. Reduction of leaf nitrogen content under elevated O 3 conditions was observed only in Siebold's beech in October. These results indicated that the decrease in the efficiency of photosynthetic nitrogen use is in an earlier step in O 3 -induced decline of photosynthesis in Siebold's beech and Japanese oak. Based on these results, we emphasize the importance of integration of O 3 effects into the conventional estimation of V cmax and J max from leaf nitrogen content for evaluating canopy photosynthesis under current and future elevated O 3 conditions., (Copyright © 2018. Published by Elsevier B.V.)

Background: Small GTPases act as molecular switches that regulate various plant responses such as disease resistance, pollen tube growth, root hair development, cell wall patterning and hormone responses. Thus, to monitor their activation status within plant cells is believed to be the key step in understanding their roles., Results: We have established a plant version of a Förster resonance energy transfer (FRET) probe called Ras and interacting protein chimeric unit (Raichu) that can successfully monitor activation of the rice small GTPase OsRac1 during various defence responses in cells. Here, we describe the protocol for visualizing spatiotemporal activity of plant Rac/ROP GTPase in living plant cells, transfection of rice protoplasts with Raichu - OsRac1 and acquisition of FRET images., Conclusions: Our protocol should be adaptable for monitoring activation for other plant small GTPases and protein-protein interactions for other FRET sensors in various plant cells.

Purpose: Digitization of clinical observation is necessary for assessing the severity of superior limbic keratoconjunctivitis (SLK). This study aimed to use a novel quantitative marker to examine hyperemia in patients with SLK., Materials and Methods: We included six eyes of six patients with both dry eye disease and SLK (SLK group) and eight eyes of eight patients with Sjögren syndrome (SS group). We simultaneously obtained the objective finding scores by using slit-lamp examination and calculated the superior hyperemia index (SHI) with an automated conjunctival hyperemia analysis software by using photographs of the anterior segment. Three objective finding scores, including papillary formation of the superior palpebral conjunctiva, superior limbal hyperemia and swelling, and superior corneal epitheliopathy, were determined. The SHI was calculated as the superior/temporal ratio of bulbar conjunctival hyperemia by using the software. Fisher's exact test was used to compare a high SHI (≥1.07) ratio between the SLK and SS groups. P-Values < 0.05 were considered statistically significant., Results: The SHI (mean ± standard deviation) in the SLK and SS groups was 1.19 ± 0.50 and 0.69 ± 0.24, respectively. The number of patients with a high SHI (≥1.07) was significantly higher in the SLK group than in the SS group (p < 0.05). The sensitivity and specificity of the SHI in the differential diagnosis between SS and SLK were 66.7% and 87.5%, respectively. An analysis of the association between the objective finding scores and SHI showed that the SHI had a tendency to indicate the severity of superior limbal hyperemia and swelling score in the SLK group., Conclusion: The SHI calculated using the automated conjunctival hyperemia analysis software could successfully quantify superior bulbar conjunctival hyperemia and may be a useful tool for the differential diagnosis between SS and SLK and for the quantitative follow-up of patients with SLK.

A cationic fluorescent nanogel thermometer based on thermo-responsive N-isopropylacrylamide and environment-sensitive benzothiadiazole was developed with a new azo compound bearing imidazolium rings as the first cationic radical initiator. This cationic fluorescent nanogel thermometer showed an excellent ability to enter live mammalian cells in a short incubation period (10 min), a high sensitivity to temperature variations in live cells (temperature resolution of 0.02-0.84 °C in the range 20-40 °C), and remarkable non-cytotoxicity, which permitted ordinary cell proliferation and even differentiation of primary cultured cells., (© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)