Investigation of Neuromodulatory Effect of Anodal Cerebellar Transcranial Direct Current Stimulation on the Primary Motor Cortex Using Functional Near-Infrared Spectroscopy.

Cerebellar brain inhibition (CBI), a neural connection between the cerebellum and primary motor cortex (M1), has been researched as a target pathway for neuromodulation to improve clinical outcomes in various neurological diseases. However, conflicting results of anodal cerebellar transcranial direct current stimulation (acb-tDCS) on M1 excitability indicate that additional investigation is required to examine its precise effect. This study aimed to gather evidence of the neuromodulatory effect of acb-tDCS on the M1 using functional near-infrared spectroscopy (fNIRS). Sixteen healthy participants were included in this cross-over study. Participants received real and sham acb-tDCS randomly, with a minimum 1-week washout period between them. The anode and cathode were placed on the right cerebellum and the right buccinator muscle, respectively. Stimulation lasted 20 min at an intensity of 2 mA, and fNIRS data were recorded for 42 min (including a 4-min baseline before stimulation and an 18-min post-stimulation duration) using eight channels attached bilaterally on the M1. acb-tDCS induced a significant decrease in oxyhemoglobin (HbO) concentration (inhibitory effect) in the left (contralateral) M1, whereas it induced a significant increase in HbO concentration (excitatory effect) in the right (ipsilateral) M1 compared to sham tDCS during (p < 0.05) and after stimulation (p < 0.01) in a group level analysis. At the individual level, variations in response to acb-tDCS were observed. Our findings demonstrate the neuromodulatory effects of acb-tDCS on the bilateral M1 in terms of neuronal hemodynamics.

Calcium Induces the Cleavage of NopA and Regulates the Expression of Nodulation Genes and Secretion of T3SS Effectors in Sinorhizobium fredii NGR234.

The type III secretion system (T3SS) is a key factor for the symbiosis between rhizobia and legumes. In this study, we investigated the effect of calcium on the expression and secretion of T3SS effectors (T3Es) in Sinorhizobium fredii NGR234, a broad host range rhizobial strain. We performed RNA-Seq analysis of NGR234 grown in the presence of apigenin, calcium, and apigenin plus calcium and compared it with NGR234 grown in the absence of calcium and apigenin. Calcium treatment resulted in a differential expression of 65 genes, most of which are involved in the transport or metabolism of amino acids and carbohydrates. Calcium had a pronounced effect on the transcription of a gene (NGR_b22780) that encodes a putative transmembrane protein, exhibiting a 17-fold change when compared to NGR234 cells grown in the absence of calcium. Calcium upregulated the expression of several sugar transporters, permeases, aminotransferases, and oxidoreductases. Interestingly, calcium downregulated the expression of nodABC, genes that are required for the synthesis of nod factors. A gene encoding a putative outer membrane protein (OmpW) implicated in antibiotic resistance and membrane integrity was also repressed by calcium. We also observed that calcium reduced the production of nodulation outer proteins (T3Es), especially NopA, the main subunit of the T3SS pilus. Additionally, calcium mediated the cleavage of NopA into two smaller isoforms, which might affect the secretion of other T3Es and the symbiotic establishment. Our findings suggest that calcium regulates the T3SS at a post-transcriptional level and provides new insights into the role of calcium in rhizobia-legume interactions.

Bowman-Birk Inhibitor Mutants of Soybean Generated by CRISPR-Cas9 Reveal Drastic Reductions in Trypsin and Chymotrypsin Inhibitor Activities.

Despite the high quality of soybean protein, raw soybeans and soybean meal cannot be directly included in animal feed mixtures due to the presence of Kunitz (KTi) and Bowman-Birk protease inhibitors (BBis), which reduces animal productivity. Heat treatment can substantially inactivate trypsin and chymotrypsin inhibitors (BBis), but such treatment is energy-intensive, adds expense, and negatively impacts the quality of seed proteins. As an alternative approach, we have employed CRISPR/Cas9 gene editing to create mutations in BBi genes to drastically lower the protease inhibitor content in soybean seed. Agrobacterium-mediated transformation was used to generate several stable transgenic soybean events. These independent CRISPR/Cas9 events were examined in comparison to wild-type plants using Sanger sequencing, proteomic analysis, trypsin/chymotrypsin inhibitor activity assays, and qRT-PCR. Collectively, our results demonstrate the creation of an allelic series of loss-of-function mutations affecting the major BBi gene in soybean. Mutations in two of the highly expressed seed-specific BBi genes lead to substantial reductions in both trypsin and chymotrypsin inhibitor activities.

AXL+SIGLEC6+ dendritic cells in cerebrospinal fluid and brain tissues of patients with autoimmune inflammatory demyelinating disease of CNS.

Inflammatory demyelinating disease of the CNS (IDD) is a heterogeneous group of autoimmune diseases, and multiple sclerosis is the most common type. Dendritic cells (DCs), major antigen-presenting cells, have been proposed to play a central role in the pathogenesis of IDD. The AXL+SIGLEC6+ DC (ASDC) has been only recently identified in humans and has a high capability of T cell activation. Nevertheless, its contribution to CNS autoimmunity remains still obscure. Here, we aimed to identify the ASDC in diverse sample types from IDD patients and experimental autoimmune encephalomyelitis (EAE). A detailed analysis of DC subpopulations using single-cell transcriptomics for the paired cerebrospinal fluid (CSF) and blood samples of IDD patients (total n = 9) revealed that three subtypes of DCs (ASDCs, ACY3+ DCs, and LAMP3+ DCs) were overrepresented in CSF compared with their paired blood. Among these DCs, ASDCs were also more abundant in CSF of IDD patients than in controls, manifesting poly-adhesional and stimulatory characteristics. In the brain biopsied tissues of IDD patients, obtained at the acute attack of disease, ASDC were also frequently found in close contact with T cells. Lastly, the frequency of ASDC was found to be temporally more abundant in acute attack of disease both in CSF samples of IDD patients and in tissues of EAE, an animal model for CNS autoimmunity. Our analysis suggests that the ASDC might be involved in the pathogenesis of CNS autoimmunity.

Comprehensiveness of cardiac rehabilitation program in Korea: a nation-wide survey result.

BACKGROUND: Cardiac rehabilitation (CR) is an essential component in secondary prevention of cardiovascular diseases. Current guidelines recommend that the program should be comprehensive including multidisciplinary behavioral intervention, not only exercise training. While the utilization of CR is gradually increasing, the comprehensiveness of the program has not been systemically evaluated in Korea. METHODS: During the year 2020, nation-wide survey was done to evaluate the current status of CR in Korea. Survey was done by web-based structured questionnaire. Survey was requested to 164 hospitals performing percutaneous coronary intervention. RESULTS: Among 164 hospitals, 47 (28.7%) hospitals had CR programs. In hospitals with CR, multidisciplinary intervention other than exercise-based program was provided only partially: nutritional counseling (63%), vocational counseling for return to work (39%), stress management (31%), psychological evaluation (18%). Personnel for CR was commonly not dedicated to the program or even absent: (percentage of dedicated, concurrent with other work, absent) physical therapist (59, 41, 0%), nurse (31, 69, 0%), dietician (6, 65, 29%), clinical psychologist (0, 37, 63%). CONCLUSION: Comprehensiveness of CR in Korea is suboptimal and human resource for it is poorly disposed. More awareness of current status by both clinicians and health policy makers is needed and insurance reimbursement for educational program should be improved.

Incidence of dysphagia requiring medical attention in various types of cancers: A nationwide population-based cohort study.

PURPOSE: Most previous reports on dysphagia in cancer have focused on specific cancer types, particularly head and neck cancer. Therefore, we aimed to investigate the incidence of dysphagia in patients with various cancers using a nationwide database in South Korea. METHODS: This was a retrospective cohort study using the National Health Insurance Service database. Claim codes were used for the selection criteria and operational definitions. The total population data from 2010 to 2015 were extracted. The crude incidence of dysphagia was calculated per 1,000 person-years. The multivariate adjusted Cox proportional hazards regression was used to determine the effects of different cancers on the incidence of dysphagia. RESULTS: People with cancer had a lower income and suffered from a higher risk of comorbidities compared to people without cancer. The risk of dysphagia increased in all types of cancers, particularly in the oral cavity and pharynx (hazard ratio [HR]: 20.65, 95% confidence interval [CI]: 17.73-24.06), esophagus (HR: 18.25, 95% CI: 15.66-21.26), larynx (HR: 12.87, 95% CI: 10.33-16.02), and central nervous system (HR: 12.42, 95% CI: 10.33-14.94). CONCLUSIONS: The risk of dysphagia was significantly higher in the cancer group than in the non-cancer group. As the survival of cancer patients is improving with the development of new treatments, more attention should be paid to dysphagia in the management of cancer. Prompt and appropriate multidisciplinary interventions for dysphagia are necessary to improve the recovery and quality of life in cancer patients.

Dysphagia Requiring Medical Attention in Parkinson's Disease: A Korean Population-Based Study.

BACKGROUND: Patients with Parkinson's disease (PD) experience both motor and non-motor symptoms, including dysphagia. Although PD is closely associated with dysphagia, the prevalence or risk of dysphagia in PD is unclear, especially in Asian countries. METHODS: The prevalence of PD and dysphagia with PD in the general population was analyzed using the Korean National Health Insurance Service (NHIS) database. The prevalence per 100,000 persons of PD and dysphagia with PD from 2006 to 2015 was analyzed in the general population aged ≥ 40 years. Patients newly diagnosed with PD between 2010 and 2015 were compared with those without PD. RESULTS: The prevalence of PD and dysphagia in patients with PD increased continuously during the study period and was highest in the ninth decade of life. The percentage of patients with dysphagia in patients with PD increased with age. Patients with PD showed an adjusted hazard ratio of 3.132 (2.955-3.320) for dysphagia compared to those without PD. CONCLUSION: This nationwide study showed increasing trends in the prevalence of PD and dysphagia among patients with PD in Korea between 2006 and 2015. The risk of dysphagia was three times higher in patients with PD than that in those without PD, highlighting the importance of providing particular attention.

The Berlin Bimanual Test for Tetraplegia (BeBiTT): development, psychometric properties, and sensitivity to change in assistive hand exoskeleton application.

BACKGROUND: Assistive hand exoskeletons are promising tools to restore hand function after cervical spinal cord injury (SCI) but assessing their specific impact on bimanual hand and arm function is limited due to lack of reliable and valid clinical tests. Here, we introduce the Berlin Bimanual Test for Tetraplegia (BeBiTT) and demonstrate its psychometric properties and sensitivity to assistive hand exoskeleton-related improvements in bimanual task performance. METHODS: Fourteen study participants with subacute cervical SCI performed the BeBiTT unassisted (baseline). Thereafter, participants repeated the BeBiTT while wearing a brain/neural hand exoskeleton (B/NHE) (intervention). Online control of the B/NHE was established via a hybrid sensorimotor rhythm-based brain-computer interface (BCI) translating electroencephalographic (EEG) and electrooculographic (EOG) signals into open/close commands. For reliability assessment, BeBiTT scores were obtained by four independent observers. Besides internal consistency analysis, construct validity was assessed by correlating baseline BeBiTT scores with the Spinal Cord Independence Measure III (SCIM III) and Quadriplegia Index of Function (QIF). Sensitivity to differences in bimanual task performance was assessed with a bootstrapped paired t-test. RESULTS: The BeBiTT showed excellent interrater reliability (intraclass correlation coefficients > 0.9) and internal consistency (α = 0.91). Validity of the BeBiTT was evidenced by strong correlations between BeBiTT scores and SCIM III as well as QIF. Wearing a B/NHE (intervention) improved the BeBiTT score significantly (p < 0.05) with high effect size (d = 1.063), documenting high sensitivity to intervention-related differences in bimanual task performance. CONCLUSION: The BeBiTT is a reliable and valid test for evaluating bimanual task performance in persons with tetraplegia, suitable to assess the impact of assistive hand exoskeletons on bimanual function.

Editor's Choice - Effect of Physical Activity and Tobacco Use on Mortality and Morbidity in Patients with Peripheral Arterial Disease After Revascularisation: A Korean Nationwide Population Based Cohort Study.

OBJECTIVE: To investigate the effects of physical activity (PA) and tobacco use on adverse clinical outcomes after revascularisation for peripheral arterial disease (PAD) in the Western Pacific region, where PAD cases and tobacco use are among the highest in the world. METHODS: This was a retrospective cohort study using the Korean National Health Insurance Service (NHIS) database and included patients who had received revascularisation for PAD between 2010 and 2015. They were categorised as active or inactive based on the number of days per week they engaged in PA and as current or non-tobacco users (self report). The primary outcome was all cause mortality. Secondary outcomes included major adverse outcome (a composite of all cause mortality, myocardial infarction, and stroke) and major adverse limb event (MALE, a composite of amputation and recurrent revascularisation). RESULTS: The relatively healthy cohort comprised 8 324 patients (mean age 64.7 years; 76.9% male) following revascularisation for PAD. Among them, 32.7% were inactive and 26.4% were tobacco users. Active patients had better outcomes than inactive patients (all cause mortality adjusted hazard ratio [adjHR] 0.766; 95% CI 0.685 - 0.855, major adverse outcome adjHR 0.795; 95% CI 0.719 - 0.878, MALE adjHR 0.858; 95% CI 0.773 - 0.953). Tobacco users had poorer outcomes than non-users (all cause mortality adjHR 1.279; 95% CI 1.124 - 1.456, major adverse outcome adjHR 1.263; 95% CI 1.124 - 1.418, MALE adjHR 1.291; 95% CI 1.143 - 1.458). CONCLUSION: Even after receiving revascularisation for PAD, a sizable proportion of patients were physically inactive and used tobacco, leading to adverse clinical outcomes such as death, cardiovascular morbidity, and amputation in Korea. These modifiable risk factors should be addressed systematically, and a comprehensive approach including supervised exercise programmes and tobacco cessation is needed in patients with PAD.

EDCs trigger immune-neurotransmitter related gene expression, and cause histological damage in sensitive mud crab Macrophthalmus japonicus gills and hepatopancreas.

Endocrine-disrupting chemicals (EDCs), distributed at various concentrations in freshwater and marine ecosystems, affect the survival, reproduction, and behavior of wide ranges organisms. Most toxicology studies on EDCs have focused on the endocrine system of invertebrates, and research on invertebrate neurotransmitters is limited. In the present study, we investigated the expression of Macrophthalmus japonicus genes encoding γ-aminobutyric acid transporter subtype 2 (GAT-2) and glutamine synthetase (GS), which play important roles as neurotransmitters at synapses. We observed differences in the mRNA expression levels of GAT-2 and GS as well as histological changes in various tissues after exposure to bisphenol-A (BPA) and di-(2-ethylhexyl) phthalate (DEHP). The amino acid sequences of M. japonicus GAT-2 and GS formed separate branches in crustaceans, fish, insects, and mammals. M. japonicus GAT-2 and GS expression levels were highest in the gills, hepatopancreas, and stomach, and showed different between DEHP or BPA treatments. In particular, hepatopancreas GS expression on Day 1, the first step in the presynaptic process, was upregulated after BPA and DEHP exposure, while GAT-2, sequential step in the presynaptic process, was significantly elevated only in DEHP. After BPA treatments, gill GS expression was increased at all concentrations, whereas GAT-2 expression was overall down regulations. In contrast, in DEHP treatment groups hepatopancreatic GS and GAT-2 expression at Day 1 was only significantly higher and all groups including gill GS and GAT-2 expression were downregulation. Histological changes in the gills and hepatopancreas were observed in a concentration-dependent manner. Accordingly, BPA and DEHP exposure in crabs could be stimulate neurotransmitter gene expression and alter the morphological structure of gill and hepatopancreas.

A survey of the perceptions of barriers to and facilitators of cardiac rehabilitation in healthcare providers and policy stakeholders.

BACKGROUND: Cardiac rehabilitation (CR) is a prognostic management strategy to help patients with CVD achieve a good quality of life and lower the rates of recurrence, readmission, and premature death from disease. Globally, cardiac rehabilitation is poorly established in hospitals and communities. Hence, this study aimed to investigate the discrepancies in the perceptions of the need for CR programs and relevant health policies between directors of hospitals and health policy personnel in South Korea to shed light on the status and to establish practically superior and effective strategies to promote CR in South Korea. METHODS: We sent a questionnaire to 592 public health policy managers and directors of selected hospitals, 132 of whom returned a completed questionnaire (response rate: 22.3%). The participants were categorized into five types of organizations depending on their practice of PCI (Percutaneous Coronary Intervention), establishment of cardiac rehabilitation, director of hospital, and government's policy makers. Differences in the opinions between directors of hospitals that perform/do not perform PCI, directors of hospitals with/without cardiac rehabilitation, and between hospital directors and health policy makers were analyzed. RESULTS: Responses about targeting diseases for cardiac rehabilitation, patients' roles in cardiac rehabilitation, hospitals' roles in cardiac rehabilitation, and governmental health policies' roles in cardiac rehabilitation were more positive among hospitals that perform PCI than those that do not. Responses to questions about the effectiveness of cardiac rehabilitation and hospitals' roles in cardiac rehabilitation tended to be more positive in hospitals with cardiac rehabilitation than in those without. Hospital directors responded more positively to questions about targeting diseases for cardiac rehabilitation and governmental health policies' roles in cardiac rehabilitation than policy makers, and both hospitals and public organizations provided negative responses to the question about patients' roles in cardiac rehabilitation. Responses to questions about targeting diseases for cardiac rehabilitation, patients' roles in cardiac rehabilitation, and governmental health policies' roles in cardiac rehabilitation were more positive in hospitals that perform PCI than those that do not and public organizations. CONCLUSIONS: Hospitals must ensure timely referral, provide education, and promote the need for cardiac rehabilitation. In addition, governmental socioeconomic support is needed in a varity of aspects.

Evaluation of Current Resources Available for Community-Based Cardiac Rehabilitation in Korea: A Nationwide Survey Study.

BACKGROUND: In Korea, the actual distribution of cardiac rehabilitation (CR) to the clinical field is insufficient due to the many barriers for cardiovascular patients to participate in CR. Community-based CR is a useful alternative to overcome these obstacles. Through a nationwide survey, we investigated the possibility of regional medical and public health management institutes which can be in charge of community-based CR in Korea. METHODS: The questionnaires on recognition of CR and current available resources in health-related institutions were developed with reference to the CR evaluation tools of York University and the International Council of Cardiovascular Prevention and Rehabilitation. The questionnaires were sent to regional public and private medical institutions and public health management institutions. RESULTS: In total, 2,267 questionnaires were sent to 1,186 institutions. There were 241 and 242 responses from 173 and 179 regional private and public medical institutions, respectively. And a total of 244 responses were gathered from 180 public health management institutions. Although many institutions were equipped with the necessary facilities for exercise training, there were few patient-monitoring systems during exercise. Most institutions were aware of the need for CR, but were burdened with the cost of establishing personnel and facilities to operate CR. CONCLUSION: Most regional medical, and public health management institutions in Korea are unprepared for the implementation of community-based CR programs. To encourage the utilization of such, there should be efforts to establish a national consensus.

Comparison of Factors Associated With Direct Versus Transferred-in Admission to Government-Designated Regional Centers Between Acute Ischemic Stroke and Myocardial Infarction in Korea.

BACKGROUND: There has been no comparison of the determinants of admission route between acute ischemic stroke (AIS) and acute myocardial infarction (AMI). We examined whether factors associated with direct versus transferred-in admission to regional cardiocerebrovascular centers (RCVCs) differed between AIS and AMI. METHODS: Using a nationwide RCVC registry, we identified consecutive patients presenting with AMI and AIS between July 2016 and December 2018. We explored factors associated with direct admission to RCVCs in patients with AIS and AMI and examined whether those associations differed between AIS and AMI, including interaction terms between each factor and disease type in multivariable models. To explore the influence of emergency medical service (EMS) paramedics on hospital selection, stratified analyses according to use of EMS were also performed. RESULTS: Among the 17,897 and 8,927 AIS and AMI patients, 66.6% and 48.2% were directly admitted to RCVCs, respectively. Multivariable analysis showed that previous coronary heart disease, prehospital awareness, higher education level, and EMS use increased the odds of direct admission to RCVCs, but the odds ratio (OR) was different between AIS and AMI (for the first 3 factors, AMI > AIS; for EMS use, AMI < AIS). EMS use was the single most important factor for both AIS and AMI (OR, 4.72 vs. 3.90). Hypertension and hyperlipidemia increased, while living alone decreased the odds of direct admission only in AMI; additionally, age (65-74 years), previous stroke, and presentation during non-working hours increased the odds only in AIS. EMS use weakened the associations between direct admission and most factors in both AIS and AMI. CONCLUSIONS: Various patient factors were differentially associated with direct admission to RCVCs between AIS and AMI. Public education for symptom awareness and use of EMS is essential in optimizing the transportation and hospitalization of patients with AMI and AIS.

Interactions of gene expression, alternative splicing, and DNA methylation in determining nodule identity.

Soybean nodulation is a highly controlled process that involves complex gene regulation at both transcriptional and post-transcriptional levels. In the present study, we profiled gene expression changes, alternative splicing events, and DNA methylation patterns during nodule formation, development, and senescence. The transcriptome data uncovered key transcription patterns of nodule development that included 9669 core genes and 7302 stage-specific genes. Alternative splicing analysis uncovered a total of 2323 genes that undergo alternative splicing events in at least one nodule developmental stage, with activation of exon skipping and repression of intron retention being the most common splicing events in nodules compared to roots. Approximately 40% of the differentially spliced genes were also differentially expressed at the same nodule developmental stage, implying a substantial association between gene expression and alternative splicing. Genome-wide-DNA methylation analysis revealed dynamic changes in nodule methylomes that were specific to each nodule stage, occurred in a sequence-specific manner, and impacted the expression of 1864 genes. An attractive hypothesis raised by our data is that increased DNA methylation may contribute to the efficiency of alternative splicing. Together, our results provide intriguing insights into the associations between gene expression, alternative splicing, and DNA methylation that may shape transcriptome complexity and proteome specificity in developing soybean nodules.

Improved cortical activity and reduced gait asymmetry during poststroke self-paced walking rehabilitation.

BACKGROUND: For patients with gait impairment due to neurological disorders, body weight-supported treadmill training (BWSTT) has been widely used for gait rehabilitation. On a conventional (passive) treadmill that runs at a constant speed, however, the level of patient engagement and cortical activity decreased compared with gait training on the ground. To increase the level of cognitive engagement and brain activity during gait rehabilitation, a self-paced (active) treadmill is introduced to allow patients to actively control walking speed, as with overground walking. METHODS: To validate the effects of self-paced treadmill walking on cortical activities, this paper presents a clinical test with stroke survivors. We hypothesized that cortical activities on the affected side of the brain would also increase during active walking because patients have to match the target walking speed with the affected lower limbs. Thus, asymmetric gait patterns such as limping or hobbling might also decrease during active walking. RESULTS: Although the clinical test was conducted in a short period, the patients showed higher cognitive engagement, improved brain activities assessed by electroencephalography (EEG), and decreased gait asymmetry with the self-paced treadmill. As expected, increases in the spectral power of the low γ and ß bands in the prefrontal cortex (PFC), premotor cortex (PMC), and supramarginal gyrus (SG) were found, which are possibly related to processing sensory data and planning voluntary movements. In addition, these changes in cortical activities were also found with the affected lower limbs during the swing phase. Since our treadmill controller tracked the swing speed of the leg to control walking speed, such results imply that subjects made substantial effort to control their affected legs in the swing phase to match the target walking speed. CONCLUSIONS: The patients also showed reduced gait asymmetry patterns. Based on the results, the self-paced gait training system has the potential to train the symmetric gait and to promote the related cortical activities after stroke. Trial registration Not applicable.

An artificial neural-network approach to identify motor hotspot for upper-limb based on electroencephalography: a proof-of-concept study.

BACKGROUND: To apply transcranial electrical stimulation (tES) to the motor cortex, motor hotspots are generally identified using motor evoked potentials by transcranial magnetic stimulation (TMS). The objective of this study is to validate the feasibility of a novel electroencephalography (EEG)-based motor-hotspot-identification approach using a machine learning technique as a potential alternative to TMS. METHODS: EEG data were measured using 63 channels from thirty subjects as they performed a simple finger tapping task. Power spectral densities of the EEG data were extracted from six frequency bands (delta, theta, alpha, beta, gamma, and full) and were independently used to train and test an artificial neural network for motor hotspot identification. The 3D coordinate information of individual motor hotspots identified by TMS were quantitatively compared with those estimated by our EEG-based motor-hotspot-identification approach to assess its feasibility. RESULTS: The minimum mean error distance between the motor hotspot locations identified by TMS and our proposed motor-hotspot-identification approach was 0.22 ± 0.03 cm, demonstrating the proof-of-concept of our proposed EEG-based approach. A mean error distance of 1.32 ± 0.15 cm was measured when using only nine channels attached to the middle of the motor cortex, showing the possibility of practically using the proposed motor-hotspot-identification approach based on a relatively small number of EEG channels. CONCLUSION: We demonstrated the feasibility of our novel EEG-based motor-hotspot-identification method. It is expected that our approach can be used as an alternative to TMS for motor hotspot identification. In particular, its usability would significantly increase when using a recently developed portable tES device integrated with an EEG device.

Prevalence of Worsening Problems Using Post-Stroke Checklist and Associations with Quality of Life in Patients with Stroke.

BACKGROUND AND AIM: A post-stroke checklist was developed to improve the standard of long-term post-stroke care and health-related quality of life. We aimed to investigate the prevalence of worsening problems using a post-stroke checklist at 3, 6, and 12 months post-stroke and their associations with health-related quality of life in patients with stroke. METHODS: In stroke patients admitted between June 2014 and December 2015, post-stroke checklist and EuroQol-5D three level were assessed at post-stroke 3 (n=181), 6 (n=175), and 12 months (n=89). The prevalence of worsening problems and its association with EQ-5D index at post-stroke 3 and 6 months were analyzed. RESULTS: The most frequently and continuously identified worsening problems were mood disturbances (reported by 8.8%, 16.0% and 13.5% of patients at 3, 6, and 12 months post-stroke, respectively). Worsening mobility was significantly associated with worse EQ-5D index at post-stroke 3 months (ß= -0.583; 95% confidence interval [CI], -1.045 to -0.120; P=0.014). The worsening of mobility and communication was significantly associated with worse EQ-5D index at post-stroke 6 months (mobility: ß= -0.170; 95% confidence interval [CI], -0.305 to -0.034; P=0.014, communication: ß=-0.164; 95% confidence interval [CI], -0.309 to -0.020; P=0.026). CONCLUSIONS: A post-stroke checklist may be useful for detection of various subjective worsening problems during serial clinical follow-up after stroke. Appropriate rehabilitation and management strategy to solve the identified problems could improve the quality of life in stroke survivors.

Effect of Heat Stress on Seed Protein Composition and Ultrastructure of Protein Storage Vacuoles in the Cotyledonary Parenchyma Cells of Soybean Genotypes That Are Either Tolerant or Sensitive to Elevated Temperatures.

High growth temperatures negatively affect soybean (Glycine max (L.) Merr) yields and seed quality. Soybean plants, heat stressed during seed development, produce seed that exhibit wrinkling, discoloration, poor seed germination, and have an increased potential for incidence of pathogen infection and an overall decrease in economic value. Soybean breeders have identified a heat stress tolerant exotic landrace genotype, which has been used in traditional hybridization to generate experimental genotypes, with improved seed yield and heat tolerance. Here, we have investigated the seed protein composition and ultrastructure of cotyledonary parenchyma cells of soybean genotypes that are either susceptible or tolerant to high growth temperatures. Biochemical analyses of seed proteins isolated from heat-tolerant and heat-sensitive genotypes produced under 28/22 °C (control), 36/24 °C (moderate), and 42/26 °C (extreme) day/night temperatures revealed that the accumulation in soybean seeds of lipoxygenase, the ß-subunit of ß-conglycinin, sucrose binding protein and Bowman-Birk protease inhibitor were negatively impacted by extreme heat stress in both genotypes, but these effects were less pronounced in the heat-tolerant genotype. Western blot analysis showed elevated accumulation of heat shock proteins (HSP70 and HSP17.6) in both lines in response to elevated temperatures during seed fill. Transmission electron microscopy showed that heat stress caused dramatic structural changes in the storage parenchyma cells. Extreme heat stress disrupted the structure and the membrane integrity of protein storage vacuoles, organelles that accumulate seed storage proteins. The detachment of the plasma membrane from the cell wall (plasmolysis) was commonly observed in the cells of the sensitive line. In contrast, these structural changes were less pronounced in the tolerant genotype, even under extreme heat stress, cells, for the most part, retained their structural integrity. The results of our study demonstrate the contrasting effects of heat stress on the seed protein composition and ultrastructural alterations that contribute to the tolerant genotype's ability to tolerate high temperatures during seed development.

Endocrine-disrupting chemicals impair the innate immune prophenoloxidase system in the intertidal mud crab, Macrophthalmus japonicus.

Endocrine-disrupting chemicals (EDCs), xenobiotics that interfere with endogenous hormone function, have been studied for their impacts in aquatic environments. However, there is limited information about the potentially hazardous impact of bisphenol A (BPA) and di-(2-ethylhexyl) phthalate (DEHP) on the marine environment. The aim of this study was to investigate the effects of BPA and DEHP on the immune response of the intertidal mud crab, Macrophthalmus japonicus. In order to examine immunological responses involving the prophenoloxidase (proPO) system, mRNA transcript and activity levels of six immune-related genes, including lipopolysaccharide and ß-1,3-glucan-binding protein (LGBP), proPO, phenoloxidase (PO), peroxinectin (PE), serine protease inhibitor (Serpin), and trypsin (Tryp), were assessed in M. japonicus hepatopancreas and gills exposed to BPA or DEHP. Expression of immune genes generally decreased in M. japonicus hepatopancreas and gills exposed to all concentrations of BPA by days 4 and 7. However, at day 1, expression of Serpin and Tryp genes was significantly increased in M. japonicus hepatopancreas and gills exposed to BPA. For DEHP exposure, all genes, with the exception of Serpin, were significantly downregulated in M. japonicus gills. In the hepatopancreas, gene expression of PO, proPO, and LGBP increased at day 1, and then decreased by day 7, while mRNA expression of Serpin and Tryp exhibited up-regulation over all exposure periods. In addition, PE gene expression was upregulated in hepatopancreas at day 7 in a dose-dependent manner. Taken together, these results indicated that the crab immune responses were perturbed by exposure to BPA, and, in particular, DEHP.

Endoscopic closure of iatrogenic colon perforation using dual-channel endoscope with an endoloop and clips: methods and feasibility data (with videos).

BACKGROUND: Colon perforation is the most serious complication associated with colonoscopic procedures. We performed a novel purse-string suture technique to close the iatrogenic colonic perforation using dual-channel endoscope with an endoloop and clips. METHODS: Iatrogenic colon perforations developed during diagnostic colonoscopy referred to a tertiary hospital over 10 years were considered for this endoscopic closure. An endoloop was inserted through the left channel of the endoscope and placed around the defect. The first clip was placed at the proximal site of the defect through the other channel of the endoscope, and the endoloop was anchored on the mucosa around the defect. Then, subsequent clips were placed next to previous clips and the endoloop was fixed. After the defect was encircled by the endoloop and clips, the rim of the opening was approximated by fastening the endoloop with a purse-string technique. RESULTS: A total of 8 patients were admitted to our hospital because of iatrogenic colon perforations during diagnostic colonoscopy. Of these, 2 underwent laparoscopic surgery and 6 underwent endoscopic closure by this novel purse-string suture technique. The estimated diameters of the perforations were 20 mm. All cases were successfully treated in the endoscopy unit without sedation or general anesthesia, and recovered without any complication or subsequent operation. Abdominal pain had nearly resolved within 3 days after the procedure in all patients, and only mild peritonitis was observed. CONCLUSIONS: Iatrogenic colon perforation can be treated with a purse-string suture technique using dual-channel endoscope with an endoloop and clips. This technique can be useful for relatively large colon perforations associated with diagnostic colonoscopy.