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Taiki Miyazawa,1,2 Kiyotaka Nakagawa,1,2 Takahiro Harigae,2 Ryo Onuma,2 Fumiko Kimura,2 Tomoyuki Fujii,3 Teruo Miyazawa4,5 1Vascular Biology Laboratory, Jean Mayer USDA (United States Department of Agriculture)-Human Nutrition Research Center on Aging, Tufts University, Boston, MA, USA; 2Food and Biodynamic Chemistry Laboratory, 3Terahertz Optical & Food Engineering Laboratory, Graduate School of Agricultural Science, 4Food and Biotechnology Innovation Project, New Industry Creation Hatchery Center (NICHe), 5Food and Health Science Research Unit, Graduate School of Agricultural Science, Tohoku University, Sendai, Japan Purpose: Biodegradable nanoparticles (NPs) composed of poly(D, L-lactide-co-glycolide) (PLGA) have attracted considerable attention as delivery systems of drugs and antioxidative compounds, such as β-carotene (BC). Intravenous (IV) administration of BC-containing PLGA-NPs (BC-PLGA-NPs) coated with polysorbate 80 (PS80) has been shown to effectively deliver BC to the brain. However, the whole-body distribution profile of BC is still not clear. Therefore, we investigated the accumulation of BC in various organs, including the brain, following IV administration of PS80-coated BC-PLGA-NPs in rats.Methods: PS80-coated and uncoated BC-PLGA-NPs were prepared by solvent evaporation, and administered intravenously to Sprague Dawley rats at a BC dose of 8.5 mg/rat. Accumulation of BC in various organs (brain, heart, liver, lungs, and spleen) and blood plasma was evaluated by high performance liquid chromatography with ultraviolet (UV) detection, 1 hour after administration.Results: We prepared PS80-coated BC-PLGA-NPs with an entrapment efficiency of 14%, a particle size of 260 nm, and a zeta potential of -26 mV. Coating with PS80 was found to result in significant accumulation of BC in the lungs, rather than in the brain and other tissues. Further, plasma levels of BC in the PS80-coated BC-PLGA-NP group were much lower than those of the uncoated BC-PLGA-NP group.Conclusion: Following IV administration, PS80-coated BC-PLGA-NPs are quickly transferred from plasma circulation to the lungs, rather than the brain. Significant accumulation of BC in the lungs may be useful for health-related applications. Keywords: β-carotene, intravenous administration, nanoparticles, poly(D, L-lactide-co-glycolide) (PLGA), polysorbate 80 (PS80), tissue distribution

Inorganic polyphosphate (polyP) is a linear polymer of phosphate that plays various roles in cells, including in phosphate and metal homeostasis. Homologs of the vacuolar transporter chaperone 4 (VTC4), catalyzing polyP synthesis in many eukaryotes, are absent in red algae, which are among the earliest divergent plant lineages. We identified homologs of polyphosphate kinase 1 (PPK1), a conserved polyP synthase in bacteria, in 42 eukaryotic genomes, including 31 species detected in this study and 12 species of red algae. Phylogenetic analysis suggested that most eukaryotic PPK1 homologs originated from horizontal gene transfer from a prokaryote to a plant before the divergence of red algae and Viridiplantae. In red algae, the homologs were fused to a nucleoside diphosphate-linked moiety X (Nudix) hydrolase of the diphosphoinositol polyphosphate phosphohydrolase (DIPP) family. We characterized the fusion protein CmPPK1 in the unicellular red alga Cyanidioschyzon merolae, which has been used in studies on basic features of eukaryotes. In the knockout strain ∆CmPPK1, polyP was undetectable, suggesting a primary role for CmPPK1 in polyP synthesis. In addition, ∆CmPPK1 showed altered metal balance. Mutations in the catalytically important residues of the Nudix hydrolase domain (NHD) either increased or decreased polyP contents. Both high and low polyP NHD mutants were susceptible to phosphate deprivation, indicating that adequate NHD function is necessary for normal phosphate starvation responses. The results reveal the unique features of PPK1 in red algae and promote further investigation of polyP metabolism and functions in red algae and eukaryotic evolution., Competing Interests: Declarations. Competing interests: The authors declare no competing or financial interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

[Purpose] One-leg standing motion was measured in patients with Parkinson's disease (PD) to evaluate the association between freezing of gait (FOG) and anticipatory postural adjustments (APA). [Participants and Methods] The participants included10 healthy older individuals, seven patients with PD without FOG (PD-FOG group), and seven patients with PD and FOG (PD+FOG group). An accelerometer that was built into a smartphone was attached to the lower back of each participant, and acceleration in the mediolateral direction was measured during one-leg standing. For the mediolateral component of acceleration, the time to the maximum value in the stance direction (peak latency, PL) and the amount of displacement of the maximum value (peak magnitude, PM) were analyzed as APA features. A one-way analysis of variance (ANOVA) was used to compare PL and PM among the three groups. In addition, the APA ratio (APAr), which is the ratio of PL to PM, was compared between study cohorts. [Results] Delayed PL and decreased PM were observed in the PD+FOG group, compared with the older and PD-FOG groups. In addition, APAr levels were significantly lower in the PD+FOG group. [Conclusion] Patients with PD with FOG had greater APA impairment than those with PD without FOG., Competing Interests: None., (2024©by the Society of Physical Therapy Science. Published by IPEC Inc.)

Background: This study aimed to reveal differences in the effectiveness of rehabilitation in improving activities of daily living (ADL) in patients with acute stroke across age groups and propose age-appropriate rehabilitation strategies., Methods: This observational study analyzed nationwide administrative data of inpatients admitted to hospitals with acute stroke between April 1, 2018, and March 31, 2020. The data included the average length of daily rehabilitation sessions, weekly frequency of rehabilitation sessions, and initiation of rehabilitation within 3 days. The primary outcome was the improvement in the Barthel Index (BI) score from admission to discharge. We classified the patients based on age and analyzed improvements in ADL according to rehabilitation characteristics., Results: An increased daily rehabilitation dose was associated with improved ADL, except in patients aged <65 years (risk ratio [95% confidence interval] in the 65-74, 75-85, and ≥85 years age groups: 1.20 [1.14‒1.27], 1.21 [1.15‒1.27], and 1.43 [1.34‒1.53], respectively; all p<0.001 vs. <65 years: 1.05 [0.98‒1.12]; p=0.18). A rehabilitation frequency of seven sessions per week was associated with improved ADL in the 75-85 years and ≥85 years age groups (1.06 [1.02‒1.10] and 1.08 [1.03‒1.13], respectively; both p<0.001). The effects of initiating rehabilitation within 3 days on ADL post-admission did not differ across age groups., Conclusion: Increasing the daily dose of rehabilitation was significantly associated with improved ADL in all age groups while increasing the frequency of rehabilitation per week improved ADL in older and very old patients.

Purpose: Rehabilitation after hip fracture surgery is crucial for improving physical function. Additional rehabilitation over the weekend or after working hours is reportedly associated with improved physical function; however, this may not apply to an aging population, including patients aged > 90 years. This study aimed to investigate the association between additional weekend rehabilitation and functional outcomes in different age groups., Methods: This study analyzed a cohort of patients aged ≥ 60 years who had hip fractures and were operated on from 2010 to 2018. Data were extracted from a nationwide multicentre database. Functional outcomes at discharge were compared between patients who underwent rehabilitation on weekdays only and those who underwent rehabilitation on both weekdays and weekends. The patient groups were selected using propensity score matching analysis. Furthermore, a subgroup-analysis was conducted on patients in their 60 s, 70 s, 80 s, and 90 s., Results: A total of 390,713 patients underwent surgery during the study period. After matching, each group comprised 129,583 pairs of patients. Patients who underwent weekend rehabilitation exhibited improved physical function in transferring, walking, and stair climbing at discharge, as compared with patients who did not (odds ratio [95% confidence interval]: 1.17 [1.15-1.19], 1.17 [1.15-1.2], and 1.06 [1.03-1.08], respectively). In subgroup analysis, except for stair climbing, the positive association between weekend rehabilitation and patient function was observed across all age groups., Conclusion: Weekend rehabilitation was significantly associated with improved physical function. Given the limited healthcare resources, high-demand activities such as stair climbing may be reserved for younger age groups to optimise rehabilitation therapy., (© 2024. The Author(s), under exclusive licence to European Geriatric Medicine Society.)

Background: Previous studies have shown shorter duration of general anesthesia in smokers but it is unclear in regional anesthesia among smokers. We investigated the association between smoking status and the duration of regional anesthesia., Methods: A total of 77 patients with a mean age of 47.3 years who underwent lower extremity orthopaedic surgery under regional anesthesia between January 2021 and June 2022 were enrolled. Sixteen patients were smokers and 57 patients were non-smokers. Propensity score matching was performed to balance patient characteristics. Our primary outcome was the time to onset of motor or sensory blockade and the duration required for full recovery of motor or sensory function., Results: The time to sensory loss was 43.4 (SD 35.9) minutes in the smoking group and 39.6 (SD 31.7) minutes in the non-smoking group (p = 0.69), and the time to motor blockade was 37.0 (SD 28.4) minutes in the smoking group and 30.1 (SD 24.1) minutes in the non-smoking group (p = 0.35). The time for recovery of sensory function was 1146.7 (SD 197.8) minutes in the smoking group and 1024.6 (SD 177.9) minutes in the non-smoking group (p = 0.024). The time to recovery of motor function was 978.3 (SD 220.5) minutes in the smoking group and 1090.9 (SD 222.8) minutes in the non-smoking group (p = 0.08). The duration of sensory effect was significantly longer in the smoking group than in the non-smoking group., Conclusions: We found no significant association in the onset of regional anesthesia, but the duration of sensory blockade was significantly longer in the smoking group than in the non-smoking group. Hence, attention should be paid to the risks of the insensate limb in smokers due to prolonged sensory blockade as compared to non-smokers, rather than be concerned about delays in the onset of anesthesia., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2024 The Japanese Orthopaedic Association. Published by Elsevier B.V. All rights reserved.)

Objectives: Using smartphones, we aimed to clarify the characteristics of anticipatory postural adjustments (APA) in older adults and examine the relationship between cognitive and balance functions., Methods: The study participants were 10 young and 13 older adults. An accelerometer built into a smartphone was attached to the lower back (L5) of the participant, and acceleration in the mediolateral direction was measured using a one-leg stance (OLS). As APA features, we analyzed the time to the peak value in the stance direction (peak latency [PL]) and the amount of displacement to the peak value in the stance direction (peak magnitude [PM]). Additionally, the measured PL was divided by PM for each group to obtain the APA ratio (APAr). We investigated the relationship between the APAr and Mini-BESTest subitems., Results: Older adults showed delayed PL and decreased PM levels (p < 0.01). While in the Mini-BESTest sub-items, deductions were most common in the order of dual-task and single-leg standing, and most participants with low APAr scores were degraded in APA of sub-items. The correlation was observed between APAr and both TUG and dual-task cost (DTC) (r= -0.56, r= -0.67). According to the receiver operating characteristic curve, the APAr value was 1.71 in the older age group., Conclusions: Older adults showed delayed PL and decreased PM, and APAr was associated with cognitive and locomotor functions. By evaluating the APAr at the initiation of movement, it may be possible to distinguish the APA of the older adluts from the possible to the impossible of OLS movement., Competing Interests: Declaration of Competing Interest The 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 © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Objective: To investigate a smartphone-enabled quantitative evaluation of anticipatory postural adjustments (APA) during one-leg stance (OLS) movements among individuals with Parkinson's disease (PD)., Methods: This cross-sectional study included 10 young controls, 10 older individuals, and 13 individuals with PD. A smartphone and accelerometer were attached to the participants' lower back (L5), and the movements of the lower back toward the stance side during OLS were measured. For acceleration, the time to the peak value in the stance direction (peak latency [PL]) and the amount of displacement to the peak value in the stance direction (peak magnitude [PM]) were analyzed as APA characteristics. Additionally, the measured PL was divided by the PM for each group to obtain the APA ratio (APAr) as a new index., Results: Individuals with PD showed a delayed PL and decreased PM (vs. young controls: p = .002 for PL, p < .001 for PM) (vs. older individuals: p = .022 for PL, p = .001 for PM). The APAr clustered the young controls, older individuals, and individuals with PD. According to the receiver operating characteristic curve the APAr value was 0.95, and individuals in the PD group were identified (i.e. area under the curve: 0.98; sensitivity: 85.0%; specificity: 100%). Moreover the APAr was correlated with severity and balance ability in individuals with PD (p = .015 for NFOG-Q, p = .028 for UPDRS, p = .036 for TUG, p = .015 for Mini-BESTest, p = .018 for OLS time)., Conclusions: This smartphone-based evaluation using the APAr index was reflective of disease severity and decreased balance ability among individuals with PD. The facilitation of this measurement can help clinicians and physiotherapists quantitatively evaluate the APA of individuals with PD at laboratories and hospitals as well as in home environments.

Background: Periodic fever, aphthous stomatitis, pharyngitis, and cervical adenitis (PFAPA syndrome), and Kawasaki disease (KD) are both considered to be disorders of the innate immune system, and the potential role of inflammasome activation in the immunopathogenesis of both diseases has been previously described., Case Presentation: Herein, we report the clinical courses of three patients who presented a rare combination of PFAPA syndrome and KD. Two patients who presented KD later developed the PFAPA syndrome, of whom one developed recurrent KD 2 years after the initial diagnosis. The third patient developed KD one year after the onset of PFAPA syndrome. The presence of both of these conditions within individual patients, combined with the knowledge that inflammasome activation is involved in both PFAPA syndrome and KD, suggests a shared background of inflammatory dysregulation. To elucidate the mechanism underlying shared inflammatory dysregulation, we investigated the roles of Nod-like receptors (NLRs) and their downstream inflammasome-related genes. All the patients had a frameshift variant in CARD8 (CARD8-FS). A previous study demonstrated a higher frequency of CARD8-FS, whose product loses CARD8 activity and activates the NLRP3 inflammasome, in patients with the PFAPA syndrome. Additionally, the NLRP3 inflammasome is known to be activated in patients with KD. Together, these results suggest that the CARD8-FS variant may also be essential in KD pathogenesis. As such, we analyzed the CARD8 variants among patients with KD. However, we found no difference in the variant frequency between patients with KD and the general Japanese population., Conclusions: We report the clinical courses of three patients with a rare combination of PFAPA syndrome and KD. All the patients had the CARD8-FS variant. However, we could not find a difference in the variant frequency between patients with KD and the general Japanese population. As the frequency of KD is much higher than that of PFAPA among Japanese patients, and the cause of KD is multifactorial, it is possible that only a small portion of patients with KD harbor CARD8-FS as a causative gene., Competing Interests: The 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., (© 2024 Nakamura, Kikuchi, Sakai, Kamimura, Watanabe, Onuma, Takayama, Tamiya, Mashimo, Ebata, Hamada, Suenaga, Onouchi and Kumaki.)

While photoautotrophic organisms utilize inorganic nitrogen as the nitrogen source, heterotrophic organisms utilize organic nitrogen and thus do not generally have an inorganic nitrogen assimilation pathway. Here, we focused on the nitrogen metabolism of Rapaza viridis, a unicellular eukaryote exhibiting kleptoplasty. Although belonging to the lineage of essentially heterotrophic flagellates, R. viridis exploits the photosynthetic products of the kleptoplasts and was therefore suspected to potentially utilize inorganic nitrogen. From the transcriptome data of R. viridis, we identified gene RvNaRL, which had sequence similarity to genes encoding nitrate reductases in plants. Phylogenetic analysis revealed that RvNaRL was acquired by a horizontal gene transfer event. To verify the function of the protein product RvNaRL, we established RNAi-mediated knock-down and CRISPR-Cas9-mediated knock-out experiments for the first time in R. viridis and applied them to this gene. The RvNaRL knock-down and knock-out cells exhibited significant growth only when ammonium was supplied. However, in contrast to the wild-type cells, no substantial growth was observed when nitrate was supplied. Such arrested growth in the absence of ammonium was attributed to impaired amino acid synthesis due to the deficiency of nitrogen supply from the nitrate assimilation pathway; this in turn resulted in the accumulation of excess photosynthetic products in the form of cytosolic polysaccharide grains, as observed. These results indicate that RvNaRL is certainly involved in nitrate assimilation by R. viridis. Thus, we inferred that R. viridis achieved its advanced kleptoplasty for photoautotrophy, owing to the acquisition of nitrate assimilation via horizontal gene transfer., (© 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.)

Aims: The aim of this study was to investigate the association between additional rehabilitation at the weekend, and in-hospital mortality and complications in patients with hip fracture who underwent surgery., Methods: A retrospective cohort study was conducted in Japan using a nationwide multicentre database from April 2010 to March 2018, including 572,181 patients who had received hip fracture surgery. Propensity score matching was performed to compare patients who received additional weekend rehabilitation at the weekend in addition to rehabilitation on weekdays after the surgery (plus-weekends group), as well as those who did not receive additional rehabilitation at the weekend but did receive weekday rehabilitation (weekdays-only group). After the propensity score matching of 259,168 cases, in-hospital mortality as the primary outcome and systemic and surgical complications as the secondary outcomes were compared between the two groups., Results: The plus-weekends group was significantly associated with lower in-hospital mortality rates compared with the weekdays-only group (hazard ratio 0.86; 95% confidence interval 0.8 to 0.92; p < 0.001). Systemic complications such as acute coronary syndrome, heart failure, renal failure, and sepsis were significantly lower in the plus-weekends group, whereas urinary tract infection (UTI) and surgical complications such as surgical site infection and haematoma were significantly higher in the plus-weekends group., Conclusion: Additional weekend rehabilitation was significantly associated with lower in-hospital mortality, as well as acute coronary syndrome, heart failure, renal failure, and sepsis, but was also significantly associated with a higher risk of UTI and surgical complications. This result can facilitate the effective use of the limited rehabilitation resources at the weekend and improve the clinical awareness of specific complications. To establish more robust causal associations between additional rehabilitation over the weekend and clinical outcomes, further prospective studies or randomized controlled trials with larger sample sizes are warranted., Competing Interests: The authors declare no conflicts of interest. All authors have read and agreed to the published version of the manuscript., (© 2023 The British Editorial Society of Bone & Joint Surgery.)

[Purpose] This study aimed to evaluate the reliability and validity of measuring the range of motion of joints using a remote videoconferencing system (Zoom) and a smartphone application. [Participants and Methods] This study included 16 young and healthy adults. The participants were instructed to perform shoulder joint flexion exercises in a seated position, with automatic motions, and maintain that posture throughout the measurement. Two measurements were performed: 1) angle measurement using a three-dimensional (3D) motion analyzer, and 2) angle measurement using the videoconference software, Zoom, and a smartphone application. Intra- and inter-rater reliabilities were calculated using the intraclass correlation coefficients (ICC). The degree of agreement between the representative values of each measurer and the 3D motion analyzer was examined. [Results] ICC (1, 1) for intra-examiner reliability were 0.912 and 0.996. For the inter-rater reliability, the ICC (2, 1) was 0.945. The correlation coefficient between each examiner's value and the value of the 3D motion analyzer was 0.955 and 0.980, respectively. The Bland-Altman analysis results indicated no systematic error. [Conclusion] The method of remotely measuring joint range of motion using Zoom and a smartphone application demonstrated high reliability and validity., Competing Interests: None., (2023©by the Society of Physical Therapy Science. Published by IPEC Inc.)

[Purpose] This study aimed to investigate the reliability and validity of the quantitative evaluation of anticipatory postural adjustments using smartphones. [Participants and Methods] The study included 10 young control participants who underwent a one-legged stance with an accelerometer and a smartphone that were simultaneously attached to their lower back (L5). Acceleration was measured as the mediolateral component of the lumbar movement toward the stance side. The peak value of the time (peak latency) and the amount of displacement (peak magnitude) in the stance side direction of the lumbar acceleration were analyzed as anticipatory postural adjustment features. Intra-rater reliability was calculated for both accelerometer and smartphone measurements, while inter-rater reliability was calculated for smartphone measurements by two examiners. Validity was determined for both accelerometer and smartphone measurements. [Results] In this study, the intra-rater reliability of the peak latency and peak magnitude in accelerometer and smartphone measurements was confirmed, as was the inter-rater reliability in smartphone measurements. The intra-rater reliability was confirmed through re-testing, while the validity of the accelerometer and smartphone measurements was also confirmed. [Conclusion] The findings of this study suggest that the use of smartphones to measure anticipatory postural adjustments is highly reliable and valid, making it a useful clinical balance index. The method is simple and can be used for continuous patient monitoring., Competing Interests: None., (2023©by the Society of Physical Therapy Science. Published by IPEC Inc.)

Background: Pyriform sinus fistulas (PSFs) are rare congenital anomalies of the third or fourth brachial pouch. Dyspnea is reportedly secondary to compression by a neck mass. However, hoarseness, as the first symptom of PSF, has not yet been reported., Case Presentation: This report describes an 11-year-old girl presenting with hoarseness as the first symptom of PSF. Hoarseness occurred 2 days prior to admission. On admission, she had fever, hoarseness, and an elastic soft mass on her left anterior neck. Contrast-enhanced computed tomography of the cervical region demonstrated an abscess partially infiltrating the thyroid gland and an air pocket near the pyriform sinus. Pharyngoscopy revealed swelling of the left arytenoid region, with purulent retention. The left vocal cord was swollen but not paralyzed. Additionally, the laboratory data indicated thyrotoxicosis. Suspecting a PSF infection, parenteral treatment with cefotaxime and dexamethasone was initiated. On the following day, the hoarseness disappeared, and the fever resolved. Four weeks after onset, the thyroid hormone levels returned to the normal range, and a barium esophagogram revealed residual contrast in the left pyriform sinus, leading to a diagnosis of PSF., Conclusion: PSF presenting with hoarseness as the first symptom in patients should be considered., (© 2023. The Author(s).)

Background: Influenza C virus is a pathogen that causes acute respiratory illness in children. The clinical information about this virus is limited because of the small number of isolated viruses compared to influenza A or B viruses., Methods: A total of 60 influenza C viruses were isolated by clinical tests using cell culture methods conducted in one hospital and one clinic during the 15 years from 2006 to 2020. These 60 cases were retrospectively analyzed by comparing outpatients and inpatients. Moreover, isolated viruses were analyzed for genomic changes during the study period., Results: All were younger than 7 years, and 73% of inpatients (19 out of 26) were under 2 years of age. A significant difference was found in the frequency of pneumonia, accounting for 45% and 4% of inpatients and outpatients, respectively. Most of the viruses isolated from 2006 to 2012 belonged to the S/A sublineage of the C/Sao Paulo lineage, but three sublineage viruses, including the S/A sublineage with K190N mutation, S/V sublineage, and C/Kanagawa lineage, have cocirculated since 2014. Moreover, S/A sublineage viruses were undergoing reassortment since 2014, suggesting significant changes in the virus, both antigenically and genetically. Of the 10 strains from patients with pneumonia, 7 were in the S/A sublineage, which had circulated from 2006 to 2012., Conclusion: Infants under 2 years of age were more likely to be hospitalized with pneumonia. The genomic changes that occurred in 2014 were suggested to affect the ability of the virus to spread., Competing Interests: Declaration of Competing Interest The authors declare no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background and Objective: Accelerations, ground reaction force data, and other quantitative data obtained from kinematic analyses of gait initiation serve as measures of dyskinesia in patients with Parkinson's disease. The objective of this study was to determine whether a separate center of pressure information could reveal new characteristics of reduced anticipatory postural adjustments in patients with Parkinson's disease., Methods: Ten healthy elderly subjects and 10 patients with Parkinson's disease participated in this study. Motions at gait initiation in response to a light signal to begin the task were analyzed. Anticipatory postural adjustments were characterized using trunk acceleration data and center of pressure displacement data., Results: The separated center of pressure in patients with Parkinson's disease showed longer peak latency ( p < .01) and larger peak magnitude ( p < .01) in the stance leg than in the stepping leg. In patients with Parkinson's disease, the displacement peak latency of the stance leg center of pressure correlated negatively with the Timed Up and Go time ( r = -0.46, p < .05), while displacement peak latency of the stepping leg center of pressure correlated negatively with Unified Parkinson's Disease Rating Scale ( r =  -0.47, p < .05), and positively with Berg Balance Scale score ( r = 0.50, p < .05)., Conclusions: Patients with Parkinson's disease experience asymmetry between the stance leg and stepping leg at gait initiation. These findings may help clinicians understand the changes to motor function in patients with Parkinson's disease and suggest strategies for improved rehabilitation training.

Sexual reproduction is widespread in eukaryotes; however, only asexual reproduction has been observed in unicellular red algae, including Galdieria , which branched early in Archaeplastida. Galdieria possesses a small genome; it is polyextremophile, grows either photoautotrophically, mixotrophically, or heterotrophically, and is being developed as an industrial source of vitamins and pigments because of its high biomass productivity. Here, we show that Galdieria exhibits a sexual life cycle, alternating between cell-walled diploid and cell wall-less haploid, and that both phases can proliferate asexually. The haploid can move over surfaces and undergo self-diploidization or generate heterozygous diploids through mating. Further, we prepared the whole genome and a comparative transcriptome dataset between the diploid and haploid and developed genetic tools for the stable gene expression, gene disruption, and selectable marker recycling system using the cell wall-less haploid. The BELL/KNOX and MADS-box transcription factors, which function in haploid-to-diploid transition and development in plants, are specifically expressed in the haploid and diploid, respectively, and are involved in the haploid-to-diploid transition in Galdieria , providing information on the missing link of the sexual life cycle evolution in Archaeplastida. Four actin genes are differently involved in motility of the haploid and cytokinesis in the diploid, both of which are myosin independent and likely reflect ancestral roles of actin. We have also generated photosynthesis-deficient mutants, such as blue-colored cells, which were depleted in chlorophyll and carotenoids, for industrial pigment production. These features of Galdieria facilitate the understanding of the evolution of algae and plants and the industrial use of microalgae.

Abstract: Kiyono, R, Onuma, R, Yasaka, K, Sato, S, Yahata, K, and Nakamura, M. Effects of 5-week foam rolling intervention on range of motion and muscle stiffness. J Strength Cond Res 36(7): 1890-1895, 2022-In clinical and sports settings, foam rolling (FR) intervention is widely used to increase the range of motion (ROM). However, the chronic effects of FR on ROM and muscle stiffness are unclear. The aim of this study was to investigate the effects of 5-week FR intervention on dorsiflexion ROM (DFROM) and shear elastic modulus of the medial gastrocnemius (MG) muscle. Accordingly, 30 healthy young adults were enrolled and randomly assigned to either the FR or control group with the former receiving 90 seconds of FR thrice per week over 5 weeks. Thereafter, DFROM, passive torque at DFROM, and shear elastic modulus of the MG during passive ankle dorsiflexion were assessed using a dynamometer and ultrasonography before and after 5 weeks. Our results showed that the FR group exhibited a significant increase in DFROM (Δ = 11.7%, p < 0.05, effect size = 0.39), but no changes in passive torque at DFROM, and shear elastic modulus at 0° and 10° dorsiflexion. Moreover, significant correlations were observed between change in DFROM and change in passive torque at DFROM in the FR group (rs = 0.607, p = 0.016). These results suggested that FR was an effective method to increase DFROM given its ability to change passive torque at DFROM (stretch tolerance) without altering muscle stiffness., (Copyright © 2020 National Strength and Conditioning Association.)

Secondary loss of photosynthesis is observed across almost all plastid-bearing branches of the eukaryotic tree of life. However, genome-based insights into the transition from a phototroph into a secondary heterotroph have so far only been revealed for parasitic species. Free-living organisms can yield unique insights into the evolutionary consequence of the loss of photosynthesis, as the parasitic lifestyle requires specific adaptations to host environments. Here, we report on the diploid genome of the free-living diatom Nitzschia putrida (35 Mbp), a nonphotosynthetic osmotroph whose photosynthetic relatives contribute ca. 40% of net oceanic primary production. Comparative analyses with photosynthetic diatoms and heterotrophic algae with parasitic lifestyle revealed that a combination of gene loss, the accumulation of genes involved in organic carbon degradation, a unique secretome, and the rapid divergence of conserved gene families involved in cell wall and extracellular metabolism appear to have facilitated the lifestyle of a free-living secondary heterotroph.

Antagonist contract-relax stretching and contract-relax stretching is commonly used in sports practice and rehabilitation settings. To date, no study has compared these modalities regarding muscle stiffness and stretch tolerance. This study aimed to investigate the effects of contract-relax and antagonist contract-relax stretching on dorsiflexion range of motion (ROM), stretch tolerance, and shear elastic modulus. Forty healthy participants (24 men and 16 women) took part in the study. Participants were randomly assigned to perform either contract-relax stretching or antagonist contract-relax stretching for 2 min. Outcomes were assessed on ROM, stretch tolerance, and shear elastic modulus before and after stretching. The ROM and stretch tolerance significantly increased after both contract-relax stretching (+ 5.4 ± 5.8°, p < 0.05; + 3.5 ± 8.0 Nm, p < 0.05) and antagonist contract-relax stretching (+ 6.1 ± 4.9°, p < 0.05; + 4.2 ± 6.4 Nm, p < 0.05); however, no significant difference was found between the two groups. Alternatively, the shear elastic modulus significantly decreased after both contract-relax (-31.1 ± 22.6 kPa, p < 0.05) and antagonist contract-relax stretching (-11.1 ± 22.3 kPa, p < 0.05); however, contract-relax stretching (-41.9 ± 19.6%) was more effective than antagonist contract-relax stretching (-12.5 ± 61.6%). The results of this study suggest that contract-relax stretching instead of antagonist contract-relax stretching should be conducted to decrease muscle stiffness. However, either contract-relax or antagonist contract-relax stretching can increase ROM., Competing Interests: The 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 © 2022 Fukaya, Konrad, Sato, Kiyono, Yahata, Yasaka, Onuma, Yoshida and Nakamura.)

We performed micro-X-ray fluorescence imaging of frozen-hydrated sections of a root of Pteris vittata for the first time, to the best of our knowledge, to reveal the mechanism of arsenic (As) uptake. The As distribution was successfully visualized in cross sections of different parts of the root, which showed that (i) the major pathway of As uptake changes from symplastic to apoplastic transport in the direction of root growth, and (ii) As and K have different mobilities around the stele before xylem loading, despite their similar distributions outside the stele in the cross sections. These data can reasonably explain As reduction, axially observed around the root tip in the direction of root growth and radially observed in the endodermis in the cross sections, as a consequence of the incorporation of As into the cells or symplast of the root. In addition, previous observations of As species in the midrib can be reconciled by ascribing a reduction capacity to the root cells, which implies that As reduction mechanisms at the cellular level may be an important control on the peculiar root-to-shoot transport of As in P. vittata., (© The Author(s) 2021. Published by Oxford University Press.)

"Foam Rolling" has been used in sports settings to increase range of motion and decrease muscle stiffness without decreasing muscle strength and athletic performance. However, there has been no study investigating the acute and prolonged effect of different durations of foam rolling intervention on muscle stiffness, and the minimum foam rolling intervention duration required to decrease muscle stiffness is unclear. Therefore, the purpose of this study was to investigate the acute and prolonged effect of different durations of foam rolling intervention on ROM, muscle stiffness, and muscle strength. The 45 participants were randomly allocated to 1 of 3 groups (30 s × 1 times group vs 30 s × 3 times group vs 30 s× 10 times group). The outcome measures were dorsiflexion range of motion, shear elastic modulus of medial gastrocnemius, and muscle strength before, 2 min and 30 min after foam rolling intervention. There were no significant differences before and 2 min after foam rolling intervention in 30 s×1 time group, whereas dorsiflexion range of motion was increased in both 30 s×3 times group (p = 0.042, d = 0.26) and 30 s× 10 times group (p < 0.01, d = 0.33). However, the increase in dorsiflexion range of motion was returned to baseline value after 30 minutes in both 30 s × 3 times group and 30 s × 10 times group. In addition, there were no significant changes in shear elastic modulus and muscle strength in all groups. This study suggested that foam rolling for more than 90 s or more of foam rolling was effective in order to increase the range of motion immediately without changing muscle stiffness and muscle strength., (© Journal of Sports Science and Medicine.)

Previous studies have shown significant improvement in muscle soreness and muscle function loss after 300-s foam rolling intervention two days after intense exercise. However, this duration is assumed to be too long, so investigating the effect of short-term duration foam rolling intervention on an eccentrically-damaged muscle is needed. This study aimed to eccentrically induce muscle damage in the leg extensors, and to detect the acute effect of 90-s foam rolling on muscle soreness and muscle function of the quadriceps muscle. We enrolled 17 healthy and nonathlete male volunteers. They performed a bout of eccentric exercise of the knee extensors with the dominant leg and received 90-s foam rolling intervention of the quadriceps two days after the eccentric exercise. The dependent variables were measured before the eccentric exercise (baseline), and before (preintervention) and after foam rolling intervention (postintervention), two days after the eccentric exercise. The results show that the preintervention muscle soreness and muscle strength values were significantly increased, compared with the baseline values, whereas the postintervention values were significantly decreased, compared with the preintervention values. Furthermore, 90-s of foam rolling intervention could improve muscle soreness and muscle function loss.

This study investigated the effects of static stretching (SS) delivered with the same load but using two protocols - high-intensity and short-duration and low-intensity and long-duration - on range of motion (ROM) and muscle stiffness. A total of 18 healthy students participated in the study. They randomly performed high-intensity and short-duration (120% and 100 s) or low-intensity and long-duration (50% and 240 s) SS. Outcomes were assessed on ROM, passive torque at dorsiflexion ROM, and shear elastic modulus of the medial gastrocnemius before and after static stretching. The results showed that ROM increased significantly at post-stretching compared to that at pre-stretching in both high-intensity and short-duration [+6.1° ± 4.6° (Δ25.7 ± 19.9%)] and low-intensity and long-duration [+3.6° ± 2.3° (Δ16.0 ± 11.8%)]. Also, the ROM was significantly higher at post-stretching in high-intensity and short-duration conditions than that in low-intensity and long-duration. The passive torque at dorsiflexion ROM was significantly increased in both high-intensity and short-duration [+5.8 ± 12.8 Nm (Δ22.9 ± 40.5%)] and low-intensity and long-duration [+2.1 ± 3.4 Nm (Δ6.9 ± 10.8%)] conditions, but no significant differences were observed between both conditions. The shear elastic modulus was significantly decreased in both high-intensity and short-duration [-8.8 ± 6.1 kPa (Δ - 38.8 ± 14.5%)] and low-intensity and long-duration [-8.0 ± 12.8 kPa (Δ - 22.2 ± 33.8%)] conditions. Moreover, the relative change in shear elastic modulus in the high-intensity and short-duration SS was significantly greater than that in low-intensity and long-duration SS. Our results suggest that a higher intensity of the static stretching should be conducted to increase ROM and decrease muscle stiffness, even for a short time., (Copyright © 2020 Fukaya, Kiyono, Sato, Yahata, Yasaka, Onuma and Nakamura.)

Dinothrix paradoxa and Gymnodinium quadrilobatum are benthic dinoflagellates possessing diatom-derived tertiary plastids, so-called dinotoms. Due to the lack of available genetic information, their phylogenetic relationship remains unknown. In this study, sequencing of 18S ribosomal DNA (rDNA) and the rbc L gene from temporary cultures isolated from natural samples revealed that they are close relatives of another dinotom, Galeidinium rugatum . The morphologies of these three dinotoms differ significantly from each other; however, they share a distinctive life cycle, in which the non-motile cells without flagella are their dominant phase. Cell division occurs in this non-motile phase, while swimming cells only appear for several hours after being released from each daughter cell. Furthermore, we succeeded in isolating and establishing two novel dinotom strains, HG180 and HG204, which show a similar life cycle and are phylogenetically closely related to the aforementioned three species. The non-motile cells of strain HG180 are characterized by the possession of a hemispheroidal cell covered with numerous nodes, while those of the strain HG204 form aggregations consisting of spherical smooth-surface cells. Based on the similarity in life cycles and phylogenetic closeness, we conclude that all five species should belong to a single genus, Dinothrix , the oldest genus within this clade. We transferred Ga. rugatum and Gy. quadrilobatum to Dinothrix , and described strains HG180 and HG204 as Dinothrix phymatodea sp. nov. and Dinothrix pseudoparadoxa sp. nov., (Copyright © 2020 Yamada, Sakai, Onuma, Kroth and Horiguchi.)

Endosymbiosis of unicellular eukaryotic algae into previously nonphotosynthetic eukaryotes has established chloroplasts in several eukaryotic lineages. In addition, certain unicellular organisms in several different lineages ingest algae and utilize them as temporal chloroplasts (kleptoplasts) for weeks to months before digesting them. Among these organisms, the dinoflagellate Nusuttodinium aeruginosum ingests the cryptomonad Chroomonas sp. and enlarges the kleptoplast with the aid of the cryptomonad nucleus. To understand how the cryptomonad nucleus is remodeled in the dinoflagellate, here we examined changes in the transcriptome and ploidy of the ingested nucleus. We show that, after ingestion, genes involved in metabolism, translation, and DNA replication are upregulated while those involved in sensory systems and cell motility are downregulated. In the dinoflagellate cell, the cryptomonad nucleus undergoes polyploidization that correlates with an increase in the mRNA levels of upregulated genes. In addition, the ingested nucleus almost loses transcriptional responses to light. Because polyploidization and loss of transcriptional regulation are also known to have occurred during the establishment of endosymbiotic organelles, these changes are probably a common trend in endosymbiotic evolution. Furthermore, we show that the kleptoplast and dinoflagellate are more susceptible to high light than the free-living cryptomonad but that the ingested nucleus reduces this damage.

Metabolism, cell cycle stages, and related transcriptomes in eukaryotic algae change with the diel cycle of light availability. In the unicellular red alga Cyanidioschyzon merolae , the S and M phases occur at night. To examine how diel transcriptomic changes in metabolic pathways are related to the cell cycle and to identify all genes for which mRNA levels change depending on the cell cycle, we examined diel transcriptomic changes in C. merolae In addition, we compared transcriptomic changes between the wild type and transgenic lines, in which the cell cycle was uncoupled from the diel cycle by the depletion of either cyclin-dependent kinase A or retinoblastoma-related protein. Of 4,775 nucleus-encoded genes, the mRNA levels of 1,979 genes exhibited diel transcriptomic changes in the wild type. Of these, the periodic expression patterns of 454 genes were abolished in the transgenic lines, suggesting that the expression of these genes is dependent on cell cycle progression. The periodic expression patterns of most metabolic genes, except those involved in starch degradation and de novo deoxyribonucleotide triphosphate synthesis, were not affected in the transgenic lines, indicating that the cell cycle and transcriptomic changes in most metabolic pathways are independent of the diel cycle. Approximately 40% of the cell-cycle-dependent genes were of unknown function, and approximately 19% of these genes of unknown function are shared with the green alga Chlamydomonas reinhardtii The data set presented in this study will facilitate further studies on the cell cycle and its relationship with metabolism in eukaryotic algae., (© 2020 American Society of Plant Biologists. All Rights Reserved.)

Replication of the circular bacterial chromosome is initiated at a unique origin ( oriC ) in a DnaA-dependent manner in which replication proceeds bidirectionally from oriC to ter . The nucleotide compositions of most bacteria differ between the leading and lagging DNA strands. Thus, the chromosomal DNA sequence typically exhibits an asymmetric GC skew profile. Further, free-living bacteria without genomes encoding dnaA were unknown. Thus, a DnaA- oriC -dependent replication initiation mechanism may be essential for most bacteria. However, most cyanobacterial genomes exhibit irregular GC skew profiles. We previously found that the Synechococcus elongatus chromosome, which exhibits a regular GC skew profile, is replicated in a DnaA- oriC -dependent manner, whereas chromosomes of Synechocystis sp. PCC 6803 and Nostoc sp. PCC 7120, which exhibit an irregular GC skew profile, are replicated from multiple origins in a DnaA-independent manner. Here we investigate the variation in the mechanisms of cyanobacterial chromosome replication. We found that the genomes of certain free-living species do not encode dnaA and such species, including Cyanobacterium aponinum PCC 10605 and Geminocystis sp. NIES-3708, replicate their chromosomes from multiple origins. Synechococcus sp. PCC 7002, which is phylogenetically closely related to dnaA -lacking free-living species as well as to dnaA -encoding but DnaA- oriC -independent Synechocystis sp. PCC 6803, possesses dnaA . In Synechococcus sp. PCC 7002, dnaA was not essential and its chromosomes were replicated from a unique origin in a DnaA- oriC independent manner. Our results also suggest that loss of DnaA- oriC -dependency independently occurred multiple times during cyanobacterial evolution and raises a possibility that the loss of dnaA or loss of DnaA- oriC dependency correlated with an increase in ploidy level., (Copyright © 2020 Ohbayashi, Hirooka, Onuma, Kanesaki, Hirose, Kobayashi, Fujiwara, Furusawa and Miyagishima.)

The order Trypanosomatida has been well studied due to its pathogenicity and the unique biology of the mitochondrion. In Trypanosoma brucei , four DNA polymerases, namely PolIA, PolIB, PolIC, and PolID, related to bacterial DNA polymerase I (PolI), were shown to be localized in mitochondria experimentally. These mitochondrion-localized DNA polymerases are phylogenetically distinct from other family A DNA polymerases, such as bacterial PolI, DNA polymerase gamma (Polγ) in human and yeasts, "plant and protist organellar DNA polymerase (POP)" in diverse eukaryotes. However, the diversity of mitochondrion-localized DNA polymerases in Euglenozoa other than Trypanosomatida is poorly understood. In this study, we discovered putative mitochondrion-localized DNA polymerases in broad members of three major classes of Euglenozoa-Kinetoplastea, Diplonemea, and Euglenida-to explore the origin and evolution of trypanosomatid PolIA-D. We unveiled distinct inventories of mitochondrion-localized DNA polymerases in the three classes: (1) PolIA is ubiquitous across the three euglenozoan classes, (2) PolIB, C, and D are restricted in kinetoplastids, (3) new types of mitochondrion-localized DNA polymerases were identified in a prokinetoplastid and diplonemids, and (4) evolutionarily distinct types of POP were found in euglenids. We finally propose scenarios to explain the inventories of mitochondrion-localized DNA polymerases in Kinetoplastea, Diplonemea, and Euglenida.

Skin lesions, cataracts, and congenital anomalies have been frequently associated with inherited deficiencies in enzymes that synthesize cholesterol. Lanosterol synthase (LSS) converts (S)-2,3-epoxysqualene to lanosterol in the cholesterol biosynthesis pathway. Biallelic mutations in LSS have been reported in families with congenital cataracts and, very recently, have been reported in cases of hypotrichosis. However, it remains to be clarified whether these phenotypes are caused by LSS enzymatic deficiencies in each tissue, and disruption of LSS enzymatic activity in vivo has not yet been validated. We identified two patients with novel biallelic LSS mutations who exhibited congenital hypotrichosis and midline anomalies but did not have cataracts. We showed that the blockade of the LSS enzyme reaction occurred in the patients by measuring the (S)-2,3-epoxysqualene/lanosterol ratio in the forehead sebum, which would be a good biomarker for the diagnosis of LSS deficiency. Epidermis-specific Lss knockout mice showed neonatal lethality due to dehydration, indicating that LSS could be involved in skin barrier integrity. Tamoxifen-induced knockout of Lss in the epidermis caused hypotrichosis in adult mice. Lens-specific Lss knockout mice had cataracts. These results confirmed that LSS deficiency causes hypotrichosis and cataracts due to loss-of-function mutations in LSS in each tissue. These mouse models will lead to the elucidation of the pathophysiological mechanisms associated with disrupted LSS and to the development of therapeutic treatments for LSS deficiency., Competing Interests: The authors have declared that no competing interests exist.

Feeding on unicellular photosynthetic organisms by unicellular eukaryotes is the base of the aquatic food chain and evolutionarily led to the establishment of photosynthetic endosymbionts/organelles. Photosynthesis generates reactive oxygen species and damages cells; thus, photosynthetic organisms possess several mechanisms to cope with the stress. Here, we demonstrate that photosynthetic prey also exposes unicellular amoebozoan and excavates predators to photosynthetic oxidative stress. Upon illumination, there is a commonality in transcriptomic changes among evolutionarily distant organisms feeding on photosynthetic prey. One of the genes commonly upregulated is a horizontally transferred homolog of algal and plant genes for chlorophyll degradation/detoxification. In addition, the predators reduce their phagocytic uptake while accelerating digestion of photosynthetic prey upon illumination, reducing the number of photosynthetic cells inside the predator cells, as this also occurs in facultative endosymbiotic associations upon certain stresses. Thus, some mechanisms in predators observed here probably have been necessary for evolution of endosymbiotic associations.

A monophyletic group of dinoflagellates, called 'dinotoms', are known to possess evolutionarily intermediate plastids derived from diatoms. The diatoms maintain their nuclei, mitochondria, and the endoplasmic reticulum in addition with their plastids, while it has been observed that the host dinoflagellates retain the diatoms permanently by controlling diatom karyokinesis. Previously, we showed that dinotoms have repeatedly replaced their diatoms. Here, we show the process of replacements is at two different evolutionary stages in two closely related dinotoms, Durinskia capensis and D. kwazulunatalensis. We clarify that D. capensis is a kleptoplastic protist keeping its diatoms temporarily, only for two months. On the other hand, D. kwazulunatalensis is able to keep several diatoms permanently and exhibits unique dynamics to maintain the diatom nuclei: the nuclei change their morphologies into a complex string-shape alongside the plastids during interphase and these string-shaped nuclei then condense into multiple round nuclei when the host divides. These dynamics have been observed in other dinotoms that possess permanent diatoms, while they have never been observed in any other eukaryotes. We suggest that the establishment of this unique mechanism might be a critical step for dinotoms to be able to convert kleptoplastids into permanent plastids.

Background: Kawasaki disease (KD) is an acute, systemic vasculitis syndrome that occurs in children. The clinical symptoms and epidemiologic features of KD strongly suggest that KD is triggered by unidentified infectious agents in genetically predisposed patients. In addition, a number of studies have described the role of B cells in the development of KD. To obtain a mechanistic insight into the humoral immune response of B-lineage cells in KD patients, we examined peripheral blood antibody secreting cells (ASCs) and inhibitory immunoreceptors, immunoglobulin-like transcript (ILT)/leukocyte immunoglobulin-like receptor (LILR), on each B cell subpopulation., Methods: Eighteen Japanese KD patients and thirteen healthy control subjects were recruited for this study. Their peripheral blood mononuclear cells were examined by flow cytometry for the number of CD19 B cells, the size of each B cell subset and the expression of the inhibitory isoforms of ILT/LILR on the B cell subset., Results: The frequency of CD19CD27 ASCs was significantly increased in the acute phase of KD and reduced after high-dose intravenous immunoglobulin (IVIG) treatment. Interestingly, while ILT2/LILRB1 expression was ubiquitously observed on every B cell/ASCs subset and the level was not significantly different after IVIG, ILT3/LILRB4 (B4) was uniquely expressed on only ASCs, and its expression was significantly decreased after IVIG., Conclusions: In the acute phase of KD, the frequency of ASCs is high with augmented B4 expression, whereas it is lower with decreased B4 expression after IVIG. Further studies of B4 expression on ASCs in autoimmune and infectious diseases will be needed to confirm the significance of our findings.

Hemophagoytic lymphohistiocytosis (HLH) is a rare life-threatening disorder caused by overactivation of the immune system, associated with infections, autoimmune disorders, and malignancies. The pathological hallmark of HLH is phagocytosis of blood cells and platelets by activated macrophages and histiocytes. In this report, we describe the onset of HLH in three children, aged 2, 5 and 7 years old, during the treatment of acute focal bacterial nephritis (AFBN) with an antibiotic, piperacillin-tazobactam (PIPC-TAZ). AFBN is acute localized bacterial infection of the kidney without abscess formation. PIPC-TAZ was chosen for the treatment of AFBN, because it not only has indications for complicated urinary tract infections, but also covers most of the causative bacteria of urinary tract infections, including β-lactamase-producing Escherichia coli. The clinical courses of the three patients were similar, and they were treated with PIPC-TAZ and amikacin (AMK) for AFBN. Fever went down 2 to 5 days later, and AMK was discontinued by day 6. However, fever recurred on 13 to 15 days after introduction of PIPC-TAZ therapy, even though all of the patients had no signs of recurrence of AFBN. The clinical features and laboratory tests of two patients fulfilled the criteria of HLH, whereas the other patient had initiated therapy before fulfilling the criteria. Cessation of PIPC-TAZ combined with corticosteroid therapy improved clinical symptoms. HLH of our patients was probably induced by PIPC-TAZ, as judged by the timing of the onset of HLH and the positivity of the drug-lymphocyte stimulation test. In conclusion, prolonged antibiotic therapy with PIPC-TAZ could be a cause of HLH.

Background: In the autumn of 2015, we experienced a surge in the number of pediatric cases of wheeze in our hospital, which was suspected to be caused by enterovirus (EV)-D68 transmission in the community. Thus, we implemented an ad hoc retrospective surveillance for EV-D68., Methods: Patients <15 years of age with acute respiratory infection were eligible for inclusion in this study. All enrolled patients underwent virus detection test. Additionally, neutralization tests (NTs) were performed using the stored serum samples of the enrolled patients to compare the antigenicity of the virus isolated in this study with that isolated in 2010 and evaluate the anti-EV-D68 antibody prevalence., Results: Respiratory syncytial virus (RSV) was the most commonly detected virus (35%), followed by EV-D68 (19%) and non-EV-D68 enteroviruses/human rhinoviruses (14%). Patients with EV-D68 infection had higher median age than those with RSV infection (P < 0.05). Moreover, patients with EV-D68 infection showed a higher expiratory wheeze prevalence than those with non-EV-D68 enterovirus/rhinovirus and RSV infections. The antigenicity of the isolate from the current study was similar to the virus that circulated in 2010. At the early study phase, children in our community did not have high NT titers, but the median log NT titer increased from 1.5 to 5 over time (P < 0.05)., Conclusion: This study showed the concurrent circulation of EV-D68 with non-EV-D68 enteroviruses/rhinoviruses and RSV in infants and children in our community and captured the early stage of EV-D68 transmission.

We aimed to monitor changes in the levels of amino acid neurotransmitters or neuromodulators simultaneously at the synaptic clefts of experimental animals. We developed a method for the simultaneous determination of the levels of amino acids, such as D-Ser, Gly, and L-Glu, which were involved in neurotransmission via the N-methyl-D-aspartate (NMDA) receptor, and other protein-constituted amino acids in a rat brain microdialysis (MD) sample. We used a liquid chromatography-mass spectrometry (LC-MS)/MS device equipped with a triazole-bonded column. The determination was achieved without using stable isotope-labeled compounds. We instead used suitable amino acid analogues as internal standards (ISs). We examined various analyte-IS combinations to improve reproducibility. We found a positive correlation (r = 0.720, **p < 0.0001) between relative standard deviation (%) of the area ratio and the analyte-IS retention time differences. Using the proposed method, we were able to accurately analyze trace amounts of amino acids in MD samples using ISs that were structurally similar to the analytes. Furthermore, we observed that the peripheral administration of S-methyl-L-cysteine, which was an inhibitor of the amino acid transporter Asc-1, caused some amino acid level changes in the rat brain. The proposed LC-MS/MS method can be applied in vivo to study the effects of novel therapeutic agents with monitoring the levels of amino acid neuromodulators, such as Glu, Gly, GABA, and D-Ser, in the brain. Graphical abstract LC-MS/MS analysis of amino acid enantiomers in microdialysis samples from rat striatum using triazole-bonded stationary phase.

Lutein, a type of xanthophyll, possesses antioxidative properties that contribute to the prevention of various diseases. Preliminary screening has shown that Japanese mugwort (Artemisia princeps Pamp.) contains high amounts of lutein. In this study, we evaluated the lutein concentration in a processed mugwort product (mugwort paste). By using high-performance liquid chromatography coupled with visible light detection or mass spectrometry, the lutein concentration in mugwort paste was determined as 38 mg/100 g dry weight, which indicates that mugwort is a potentially valuable natural food source of lutein. We also investigated the effects of the manufacturing process and found that the lutein content was significantly increased by the boiling and dehydrating processes during the production of mugwort paste. Mugwort paste that is rich in lutein may therefore serve as an effective nutraceutical.

Some microalgae are adapted to extremely acidic environments in which toxic metals are present at high levels. However, little is known about how acidophilic algae evolved from their respective neutrophilic ancestors by adapting to particular acidic environments. To gain insights into this issue, we determined the draft genome sequence of the acidophilic green alga Chlamydomonas eustigma and performed comparative genome and transcriptome analyses between C eustigma and its neutrophilic relative Chlamydomonas reinhardtii The results revealed the following features in C eustigma that probably contributed to the adaptation to an acidic environment. Genes encoding heat-shock proteins and plasma membrane H + -ATPase are highly expressed in C eustigma This species has also lost fermentation pathways that acidify the cytosol and has acquired an energy shuttle and buffering system and arsenic detoxification genes through horizontal gene transfer. Moreover, the arsenic detoxification genes have been multiplied in the genome. These features have also been found in other acidophilic green and red algae, suggesting the existence of common mechanisms in the adaptation to acidic environments., Competing Interests: The authors declare no conflict of interest.

The chloroplasts of cryptophytes arose through a secondary endosymbiotic event in which a red algal endosymbiont was integrated into a previously nonphotosynthetic eukaryote. The cryptophytes retain a remnant of the endosymbiont nucleus (nucleomorph) that is replicated once in the cell cycle along with the chloroplast. To understand how the chloroplast, nucleomorph and host cell divide in a coordinated manner, we examined the expression of genes/proteins that are related to nucleomorph replication and chloroplast division as well as the timing of nuclear and nucleomorph DNA synthesis in the cryptophyte Guillardia theta. Nucleus-encoded nucleomorph HISTONE H2A mRNA specifically accumulated during the nuclear S phase. In contrast, nucleomorph-encoded genes/proteins that are related to nucleomorph replication and chloroplast division (FtsZ) are constantly expressed throughout the cell cycle. The results of this study and previous studies on chlorarachniophytes suggest that there was a common evolutionary pattern in which an endosymbiont lost its replication cycle-dependent transcription while cell-cycle-dependent transcriptional regulation of host nuclear genes came to restrict the timing of nucleomorph replication and chloroplast division.

We report an infant case of rotavirus myositis, a rare complication of rotavirus infection. Complement levels of the patient were normal when serum creatine kinase (CK) level was at its peak and then decreased when the CK level became normalized. In a previous case report of rotavirus myositis, transient decrease of serum albumin, immunoglobulin, and complement levels was reported. The authors speculated that intravascular complement activation was caused by rotavirus and resulted in the pathogenesis of myositis, although complement levels at onset were not measured by the authors. In this report, however, we demonstrate that the complement activation of our patient is a result of, rather than the cause of, skeletal muscle damage.

The unarmoured freshwater dinoflagellate Nusuttodinium (= Gymnodinium) aeruginosum retains a cryptomonad-derived kleptochloroplast and nucleus, the former of which fills the bulk of its cell volume. The paucity of studies following morphological changes to the kleptochloroplast with time make it unclear how the kleptochloroplast enlarges and why the cell ultimately loses the cryptomonad nucleus. We observed, both at the light and electron microscope level, morphological changes to the kleptochloroplast incurred by the enlargement process under culture conditions. The distribution of the cryptomonad nucleus after host cell division was also investigated. The volume of the kleptochloroplast increased more than 20-fold, within 120h of ingestion of the cryptomonad. Host cell division was not preceded by cryptomonad karyokinesis so that only one of the daughter cells inherited a cryptomonad nucleus. The fate of all daughter cells originating from a single cell through five generations was closely monitored, and this observation revealed that the cell that inherited the cryptomonad nucleus consistently possessed the largest kleptochloroplast for that generation. Therefore, this study suggests that some important cryptomonad nucleus division mechanism is lost during ingestion process, and that the cryptomonad nucleus carries important information for the enlargement of the kleptochloroplast., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

In vivo incorporation and reduction abilities of 4-carboxy-2,2,6,6-tetramethylpiperidine-1-oxyl (4-carboxy-TEMPO) (1), 3-carboxy-2,2,5,5-tetramethylpyrroline-1-oxyl (3-carboxy-dehydro-PROXYL, 3-carboxy-DPRO) (2), 4-hydroxy-TEMPO and 3-hydroxymethyl-DPRO O-β-D-glucosides (3 and 5), and newly designed forms of 6-O-(TEMPO-4-carbonyl and DPRO-3-carbonyl)-D-glucose (4 and 6) were evaluated using white radish sprouts. For each of these compounds, electron spin resonance (ESR) spectrometry was used to measure two effects: the rate of in vitro reduction via the addition of ascorbic acid; and, the rate of successful incorporation into radish sprouts for a reduction to the corresponding hydroxyl amine. DPRO-radicals 2, 5, and 6 were detected significantly more than TEMPO-radicals 1, 3, and 4 in vitro and in vivo for both experiments. Four glucose-linked nitroxide radicals were reduced faster than the glucose-non-linked ones in the in vitro experiment, but were nonetheless detected more each time in radish sprouts due to the absorbability. Glucose ester-linked radicals 4 and 6 were detected more than glycosides 3 and 5, which suggests that glucose ester-linked DPRO-radical 6 is the best for use as a spin-label probe that a plant will incorporate., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Braarudosphaera bigelowii (Prymnesiophyceae) is a coastal coccolithophore with a long fossil record, extending back to the late Cretaceous (ca. 100 Ma). A recent study revealed close phylogenetic relationships between B. bigelowii, Chrysochromulina parkeae (Prymnesiophyceae), and a prymnesiophyte that forms a symbiotic association with the nitrogen-fixing cyanobacterium UCYN-A. In order to further examine these relationships, we conducted transmission electron microscopic and molecular phylogenetic studies of B. bigelowii. TEM studies showed that, in addition to organelles, such as the nucleus, chloroplasts and mitochondria, B. bigelowii contains one or two spheroid bodies with internal lamellae. In the 18S rDNA tree of the Prymnesiophyceae, C. parkeae fell within the B. bigelowii clade, and was close to B. bigelowii Genotype III (99.89% similarity). Plastid 16S rDNA sequences obtained from B. bigelowii were close to the unidentified sequences from the oligotrophic SE Pacific Ocean (e.g. HM133411) (99.86% similarity). Bacterial16S rDNA sequences obtained from B. bigelowii were identical to the UCYN-A sequence AY621693 from Arabian Sea, and fell in the UCYN-A clade. From these results, we suggest that; 1) C. parkeae is the alternate life cycle stage of B. bigelowii sensu stricto or that of a sibling species of B. bigelowii, and 2) the spheroid body of B. bigelowii originated from endosymbiosis of the nitrogen-fixing cyanobacterium UCYN-A.

The unarmoured marine dinoflagellate Amphidinium poecilochroum and the unarmoured freshwater dinoflagellate Gymnodinium aeruginosum both belonging to the same clade, are known to possess cryptomonad-derived kleptochloroplasts. Previous studies revealed that G. aeruginosum can synchronise the division of the chloroplast with its own cell division while no simultaneous division takes place in A. poecilochroum, which is interpreted to mean that state of kleptochloroplastidy in G. aeruginosum is closer to that of the initial acquisition of the 'true chloroplast' within the lineage. Although the general ultrastructure of these two species has been reported, the changes in the kleptochloroplast with time have never been followed. We observed morphological changes in kleptochloroplasts of A. poecilochroum and G. aeruginosum following the ingestion of cryptomonad cells, using light and transmission electron microscopes. In A. poecilochroum, the cryptomonad ejectosomes, mitochondria and cytoplasm were all actively transferred into digestive vacuoles within 1h of ingestion. The chloroplasts were deformed and the cryptomonad nucleus was digested after 3h. By contrast, in G. aeruginosum, the cryptomonad cytoplasm and nucleus were retained for 24h following ingestion, and the chloroplast was substantially enlarged. These differences imply that the retention of the cryptomonad nucleus is important for the maintenance of the chloroplast., (Copyright © 2013 Elsevier GmbH. All rights reserved.)