Terminus-Selective Covalent Immobilization of Heparin on a Thermoresponsive Surface Using Click Chemistry for Efficient Binding of Basic Fibroblast Growth Factor.

Cell therapy using endothelial cells (ECs) has great potential for the treatment of congenital disorders, such as hemophilia A. Cell sheet technology utilizing a thermoresponsive culture dish is a promising approach to efficiently transplant donor cells. In this study, a new method to prepare terminus-selective heparin-immobilized thermoresponsive culture surfaces is developed to facilitate the preparation of EC sheets. Alkynes are introduced to the reducing terminus of heparin via reductive amination. Cu-catalyzed azide-alkyne cycloaddition (CuAAC) facilitates efficient immobilization of the terminus of heparin on a thermoresponsive surface, resulting in a higher amount of immobilized heparin while preserving its function. Heparin-immobilized thermoresponsive surfaces prepared using CuAAC exhibit good adhesion to human endothelial colony-forming cells (ECFCs). In addition, upon further binding to basic fibroblast growth factor (bFGF) on heparin-immobilized surfaces, increased proliferation of ECFCs on the surface is observed. The confluent ECFC monolayer cultured on bFGF-bound heparin-immobilized thermoresponsive surfaces exhibits relatively high fibronectin accumulation and cell number and detaches at 22 °C while maintaining the sheet-like structure. Because heparin has an affinity for several types of bioactive molecules, the proposed method can be applied to facilitate efficient cultures and sheet formations of various cell types.

Regulation of von Willebrand factor by ADAMTS13 ameliorates lipopolysaccharide-induced lung injury in mice.

The relationship between von Willebrand factor (VWF) and inflammation has attracted considerable attention in recent years. VWF, which is stored in the Weibel-Palade bodies (WPBs) of endothelial cells (ECs), is released from WPBs in response to inflammatory stimuli and is thought to contribute to inflammation by promoting leukocyte extravasation. In this study, lung injury model mice were produced by intratracheal injection with lipopolysaccharides. The severity of lung inflammation was evaluated in mice with different genotypes (wild-type, Vwf-/-, Adamts13-/-) and mice treated with drugs that inhibit VWF function. Lung inflammation was significantly ameliorated in Vwf-/- mice compared with wild-type mice. Furthermore, inflammation was significantly suppressed in wild-type mice treated with anti-VWF A1 antibody or recombinant human ADAMTS13 compared with the untreated control group. The underlying mechanism appears to be an increased VWF/ADAMTS13 ratio at the site of inflammation and the interaction between blood cell components, such as leukocytes and platelets, and the VWF A1 domain, which promotes leukocyte infiltration into the lung. This study suggested that ADAMTS13 protein and other VWF-targeting agents may be a novel therapeutic option for treatment of pulmonary inflammatory diseases.

Generation of functional liver sinusoidal endothelial-like cells from human bone marrow-derived mesenchymal stem cells.

Introduction: Liver sinusoidal endothelial cells (LSECs) are specialized vascular endothelial cells that play an important role in the maintenance of biological homeostasis. However, the lack of versatile human LSECs has hindered research on LSECs and development of medical technologies for liver diseases including hemophilia A. In this study, we developed a technique to induce LSEC differentiation from human bone marrow-derived mesenchymal stem cells (BM-MSCs). Methods: To induce LSECs from human BM-MSCs, cytokines and chemical compounds associated with signaling implicated in LSEC differentiation and liver development were screened. Then LSEC-related genes and proteins expression in the differentiated cells were analyzed by qPCR and flow cytometry analysis, respectively. LSEC-related functions of the differentiated cells were also examined. Results: We found that the gene expression of LSEC markers, such as LYVE1, was considerably increased by culturing human BM-MSCs with bone morphogenetic protein 4, fibroblast growth factor 8b, transforming growth factor-ß signal inhibitor, and cyclic AMP. Furthermore, the differentiated cells expressed LSEC marker proteins and clearly demonstrated LSEC-specific functions, such as the uptake of hyaluronic acid. Conclusions: Our result indicate that the functional LSEC-like cells were successfully generated from human BM-MSCs using our established protocol.

The use of infrared thermography for non-invasive detection of bleeding and musculoskeletal abnormalities in patients with hemophilia: an observational study.

BACKGROUND: In patients with hemophilia (PwH), bleeding often occurs in joints and muscles, and early detection of hemorrhage is important to prevent the onset and progression of mobility impairment. Complex-Image analysis such as ultrasonography, computed tomography, and magnetic resonance imaging are used to detect bleeding. On the other hand, no simple and rapid method to detect the active bleeding has been reported. Local inflammatory responses occur when blood leaks from damaged vessels, and the temperature at the site of active bleeding could be expected to increase in these circumstances, leading to an increase in surrounding skin temperature. Therefore, the purpose of this study was to investigate whether the measurement of skin temperature using infrared thermography (IRT) can be used as a diagnostic aid to detect active bleeding. METHODS: Fifteen PwH (from 6 to 82 years old) complaining of discomfort such as pain were examined. Thermal images were obtained simultaneously at the affected sides and comparable unaffected sides. The average skin temperature of the affected side and of the unaffected side were measured. The temperature differences were calculated by subtracting the average skin temperature at the unaffected side from the affected side. RESULTS: In eleven cases with active bleeding, the skin temperature at the affected side was more than 0.3 °C higher (0.3 °C to 1.4 °C) compared to the unaffected side. In two cases without active bleeding, there were no significant differences in skin temperature between the affected and unaffected sides. In two cases with previous rib or thumb bone fracture, the skin temperature at the affected side was 0.3 °C or 0.4 °C lower than that of the unaffected side, respectively. In two cases with active bleeding in which longitudinal evaluation was conducted, the difference in skin temperature decreased after hemostatic treatment. CONCLUSION: The analysis of skin temperature deference using IRT was a useful supportive tool to readily assess musculoskeletal abnormalities and bleeding in PwH as well as to determine the success of the hemostatic treatment.

The structure of the superior and inferior parietal lobes predicts inter-individual suitability for virtual reality.

The global virtual reality (VR) market is significantly expanding and being challenged with an increased demand owing to COVID-19. Unfortunately, VR is not useful for everyone due to large interindividual variability existing in VR suitability. To understand the neurobiological basis of this variability, we obtained neural structural and functional data from the participants using 3T magnetic resonance imaging. The participants completed one of two tasks (sports training or cognitive task) using VR, which differed in the time scale (months/minutes) and domain (motor learning/attention task). Behavioral results showed that some participants improved their motor skills in the real world after 1-month training in the virtual space or obtained high scores in the 3D attention task (high suitability for VR), whereas others did not (low suitability for VR). Brain structure analysis revealed that the structural properties of the superior and inferior parietal lobes contain information that can predict an individual's suitability for VR.

Importance of Maternal Persistence in Young Children's Persistence.

Persistence of a distant goal is an important personality trait that determines academic and social success. Recent studies have shown that individual differences in persistence involve both genetic and environmental factors; however, these studies have not examined the role of maternal factors on a young children's persistence. The present study examined whether mothers' persistence is associated with persistence in children aged 3-6 years. In addition, the associations between mothers' persistence/parenting style and children's self-control/social development (prosocial behaviors and difficulties) were examined. Our results showed that maternal persistence is essential for the child's persistence. Children's self-control and social development were also associated with the mothers' persistence and parenting style. Our findings suggest that a young child's persistence may develop under the influence of a familiar adult (i.e., mother) and characterizes their social development, highlighting the importance of persistence in parenting.

Plastic frontal pole cortex structure related to individual persistence for goal achievement.

Persistent goal-directed behaviours result in achievements in many fields. However, the underlying neural mechanisms of persistence and the methods that enhance the neuroplasticity underlying persistence, remain unclear. We here demonstrate that the structural properties of the frontal pole cortex (FPC) before tasks contain information that can classify Achievers and Non-achievers (goal-directed persistence) participating in three tasks that differ in time scale (hours to months) and task domains (cognitive, language, and motor learning). We also found that most Achievers exhibit experience-dependent neuroplastic changes in the FPC after completing language and motor learning tasks. Moreover, we confirmed that a coaching strategy that used subgoals modified goal-directed persistence and increased the likelihood of becoming an Achiever. Notably, we discovered that neuroplastic changes in the FPC were facilitated by the subgoal strategy, suggesting that goal-striving, using effective coaching, optimizes the FPC for goal persistence.

Neuroanatomical Basis of Individuality in Muscle Tuning Function: Neural Correlates of Muscle Tuning.

In a conventional view of motor control, the human brain might employ an optimization principle that leads a stereotypical motor behavior which we observe as an averaged behavioral data over subjects. In this scenario, the inter-individual motor variability is considered as an observation noise. Here, we challenged this view. We considered a motor control task where the human participants manipulated arm force by coordinating shoulder and elbow torques and investigated the muscle-tuning function that represents how the brain distributed the ideal joint torques to multiple muscles. In the experimental data, we observed large inter-individual variability in the profile of a muscle-tuning function. This contradicts with a well-established optimization theory that is based on minimization of muscle energy consumption and minimization of motor variability. We then hypothesized the inter-subject differences in the structure of the motor cortical areas might be the source of the across-subjects variability of the motor behavior. This was supported by a voxel-based morphometry analysis of magnetic resonance imaging; The inter-individual variability of the muscle tuning profile was correlated with that of the gray matter volume in the premotor cortex which is ipsilateral to the used arm (i.e., right hemisphere for the right arm). This study suggests that motor individuality may originate from inter-individual variation in the cortical structure.

Reduced γ-aminobutyric acid in the superior temporal gyrus is associated with absolute pitch.

Absolute pitch (AP) is the ability to label the pitch of tones without any reference tones. Previous studies have shown that the anatomical and functional basis of AP exists in the superior temporal gyrus (STG), although the associated neurotransmitters remain unknown. We used proton magnetic resonance spectroscopy to quantify the concentration of γ-aminobutyric acid (GABA), an inhibitory neurotransmitter, in the bilateral STG of adult AP possessor and non-AP possessors. We found a significant negative correlation between AP scores and GABA concentration in the left STG. Furthermore, mean GABA concentration was significantly reduced in the AP possessors group compared to the non-AP possessors group. Our results suggest that less inhibition in the bilateral STG may enable AP possessors to associate categorical labels with tones.

Adrenergic Receptors in Individual Ventricular Myocytes: The Beta-1 and Alpha-1B Are in All Cells, the Alpha-1A Is in a Subpopulation, and the Beta-2 and Beta-3 Are Mostly Absent.

RATIONALE: It is unknown whether every ventricular myocyte expresses all 5 of the cardiac adrenergic receptors (ARs), ß1, ß2, ß3, α1A, and α1B. The ß1 and ß2 are thought to be the dominant myocyte ARs. OBJECTIVE: Quantify the 5 cardiac ARs in individual ventricular myocytes. METHODS AND RESULTS: We studied ventricular myocytes from wild-type mice, mice with α1A and α1B knockin reporters, and ß1 and ß2 knockout mice. Using individual isolated cells, we measured knockin reporters, mRNAs, signaling (phosphorylation of extracellular signal-regulated kinase and phospholamban), and contraction. We found that the ß1 and α1B were present in all myocytes. The α1A was present in 60%, with high levels in 20%. The ß2 and ß3 were detected in only ≈5% of myocytes, mostly in different cells. In intact heart, 30% of total ß-ARs were ß2 and 20% were ß3, both mainly in nonmyocytes. CONCLUSION: The dominant ventricular myocyte ARs present in all cells are the ß1 and α1B. The ß2 and ß3 are mostly absent in myocytes but are abundant in nonmyocytes. The α1A is in just over half of cells, but only 20% have high levels. Four distinct myocyte AR phenotypes are defined: 30% of cells with ß1 and α1B only; 60% that also have the α1A; and 5% each that also have the ß2 or ß3. The results raise cautions in experimental design, such as receptor overexpression in myocytes that do not express the AR normally. The data suggest new paradigms in cardiac adrenergic signaling mechanisms.

Sense of Accomplishment Is Modulated by a Proper Level of Instruction and Represented in the Brain Reward System.

Problem-solving can be facilitated with instructions or hints, which provide information about given problems. The proper amount of instruction that should be provided for learners is controversial. Research shows that tasks with intermediate difficulty induce the largest sense of accomplishment (SA), leading to an intrinsic motivation for learning. To investigate the effect of instructions, we prepared three instruction levels (No hint, Indirect hint, and Direct hint) for the same insight-problem types. We hypothesized that indirect instructions impose intermediate difficulty for each individual, thereby inducing the greatest SA per person. Based on previous neuroimaging studies that showed involvement of the bilateral caudate in learning and motivation, we expected SA to be processed in this reward system. We recruited twenty-one participants, and investigated neural activations during problem solving by functional magnetic resonance imaging (fMRI). We confirmed that the Indirect hint, which imposed intermediate difficulty, induced the largest SA among the three instruction types. Using fMRI, we showed that activations in the bilateral caudate and anterior cingulate cortex (ACC) were significantly modulated by SA. In the bilateral caudate, the indirect hint induced the largest activation, while the ACC seemed to reflect the difference between correct and incorrect trials. Importantly, such activation pattern was independent of notations (number or letter). Our results indicate that SA is represented in the reward system, and that the Indirect instruction effectively induces such sensation.

Different clinicopathological features between patients who developed early and late recurrence following surgery for renal cell carcinoma.

BACKGROUND: To evaluate the clinicopathological features and identify predictive factors in patients with early and late recurrence following initial surgery for localized renal cell carcinoma. METHODS: From April 1988 to January 2013, 486 patients without metastases at the initial diagnosis underwent either radical nephrectomy or partial nephrectomy and were followed up thereafter. Patients were divided into 3 groups; no recurrence, early recurrence (recurrence within 5 years), and late recurrence (recurrence after 5 years). Cancer-specific survival after recurrence was analyzed by using the Kaplan-Meier method. Multivariate logistic regression analysis was applied to define clinical and pathological factors correlated to early and late recurrence following surgery. RESULTS: Seventy-seven (15.8 %) and 18 (3.7 %) patients developed early and late recurrence, respectively. In multivariate logistic regression analysis, positive symptoms at diagnosis, ≥pT2, positive lymphovascular invasion, and grade 3 were independent predictive factors for early recurrence. Age at surgery and ≥pT2 were significantly correlated to late recurrence. The 5-year cancer-specific survival rate after recurrence was 72.4 and 52.9 % in the late and the early recurrence groups, respectively (P = 0.044). CONCLUSIONS: The risk factors for clinical recurrence differed according to the time that had elapsed between initial surgery and the first metastasis in patients with localized renal cell carcinoma. Our study showed age at initial surgery and the pT stage were independent predictive factors for late recurrence. Further investigation of a larger number of patients is required to predict which patients may develop recurrence in the future and to choose appropriate treatment.

Functions of the cortico-basal ganglia circuits for spoken language may extend beyond emotional-affective modulation in adults.

We support Ackermann et al.'s proposal that the cortico-basal ganglia circuits may play essential roles in the evolution of spoken language. Here we discuss further evidence indicating that the cortico-basal ganglia circuits may contribute to various aspects of spoken language including planning, learning, and controlling of speech in adulthood.

Dynamic neural network reorganization associated with second language vocabulary acquisition: a multimodal imaging study.

It remains unsettled whether human language relies exclusively on innately privileged brain structure in the left hemisphere or is more flexibly shaped through experiences, which induce neuroplastic changes in potentially relevant neural circuits. Here we show that learning of second language (L2) vocabulary and its cessation can induce bidirectional changes in the mirror-reverse of the traditional language areas. A cross-sectional study identified that gray matter volume in the inferior frontal gyrus pars opercularis (IFGop) and connectivity of the IFGop with the caudate nucleus and the superior temporal gyrus/supramarginal (STG/SMG), predominantly in the right hemisphere, were positively correlated with L2 vocabulary competence. We then implemented a cohort study involving 16 weeks of L2 training in university students. Brain structure before training did not predict the later gain in L2 ability. However, training intervention did increase IFGop volume and reorganization of white matter including the IFGop-caudate and IFGop-STG/SMG pathways in the right hemisphere. These "positive" plastic changes were correlated with the gain in L2 ability in the trained group but were not observed in the control group. We propose that the right hemispheric network can be reorganized into language-related areas through use-dependent plasticity in young adults, reflecting a repertoire of flexible reorganization of the neural substrates responding to linguistic experiences.

[Efficacy and safety of maintenance intravesical instillation therapy with bacillus Calmette-Guerin and epirubicin for non-muscle invasive bladder cancer].

The objectives of this study were to evaluate the efficacy and toxicity of maintenance intravesical instillation therapy with bacillus Calmette-Guerin (BCG) and epirubicin for non-muscle invasive bladder cancer. From April 1999 to March 2010, 27 eligible patients were enrolled in this study. After receiving one cycle of epirubicin (100 mg/100 ml) by intravesical instillation, all patients received 6 weekly alternate intravesical instillation of BCG (80 mg/50 ml) and epirubicin (50 mg/50 ml), followed by 10 monthly instillations. Among the 27 patients, 19 were men and 8 were women, with a median age of 62.4 years (range, 37-78 years). Tumor pathologic stage was pTa in 25 patients, pT1 in 2 and there were no concomitant carcinoma in situ cases. Median follow-up was 37.1 months (range, 11-82 months). The 3- year recurrence-free and progression-free survival rates were 75.3% and 96.1%, respectively. Furthermore, a high completion rate of 81.5% was achieved in this study. Adverse events of grade 3 or higher occurred in 3 patients (11.1%), 1 patient had anaphylaxis. There were no treatment-related deaths. Maintenance intravesical instillation therapy with BCG and epirubicin is a favorable therapeutic option for non-muscle invasive bladder cancer. Given the safety and benefit profile found in this study, appropriate patient selection is warranted in the future.

PET molecular imaging to investigate higher brain dysfunction in patients with neurotrauma.

INTRODUCTION: Many neurotrauma patients suffer from higher brain dysfunction even when focal brain damage is not detected with MRI. We performed functional imaging with positron emission tomography (PET) to clarify the relationship between the functional deficit and symptoms of such patients. METHODS: Patients who complain of higher brain dysfunction without apparent morphological cortical damage were recruited. Thirteen patients underwent PET study to image glucose metabolism by (18)F-FDG, and central benzodiazepine receptor (cBZD-R) by (11)C-flumazenil, together with measurement of cognition. RESULTS: Diffuse axonal injury (DAI) patients have a significant decrease in glucose metabolism and cBZD-R distribution in the cingulated cortex than normal controls. Score of cognition test was variable among patients. The degree of decreased glucose metabolism and cBZD-R in the dominant hemisphere corresponded well to the severity of cognitive disturbance. Patients with a milder type of diffuse brain injury (i.e., cerebral concussion) also showed abnormal glucose metabolism and cBZD-R distribution when they suffered from cognitive deficit. CONCLUSION: PET molecular imaging was useful for depicting the cortical dysfunction of neurotrauma patients even when morphological change was not apparent. This method may be promising in clarifying the pathophysiology of higher brain dysfunction of patients with neurotrauma, but without morphological abnormality.

Association between lower urinary tract symptoms and sexual dysfunction assessed using the core lower urinary tract symptom score and International Index of Erectile Function-5 questionnaires.

OBJECTIVE: The aim of this study is to assess the association between lower urinary tract symptoms (LUTS) and erectile dysfunction (ED) using the Core Lower Urinary Tract Symptom Score (CLSS) and the International Index of Erectile Dysfunction (IIEF)-5 questionnaires. METHODS: A total of 220 consecutive treatment-naive men completed the International Prostate Symptom Score (IPSS), CLSS, and IIEF-5 questionnaires. The clinical diagnoses were benign prostate hyperplasia (BPH, n=58), prostatic cancer (n=51), prostatitis (n=28), overactive bladder wet (OAB wet, n=16), other diagnoses (n=39), and controls (n=28). Simple statistics and predictability of low IIEF-5 score were examined. RESULTS: The total IIEF-5 score significantly correlated with both the IPSS and CLSS questionnaires (p=0.0001). Among IPSS and CLSS symptoms, factors other than daytime frequency and incomplete emptying showed significant correlation with the total IIEF-5 score. A multivariate regression model indicated nocturia and urethral pain as independent factors for low IIEF-5 scores. CONCLUSION: Urethral pain was identified as an independent factor for erectile dysfunction. The extraction of pain symptoms using CLSS questionnaire may be useful to evaluate LUTS in men with pelvic health problems.

Renoprotective effect of erythropoietin in ischemia/reperfusion injury: possible roles of the Akt/endothelial nitric oxide synthase-dependent pathway.

OBJECTIVES: It has been reported that erythropoietin protects the kidneys from ischemia/reperfusion injury. In the present study, we examined the role of Akt and endothelial nitric oxide synthase in the protective effect of erythropoietin on ischemia/reperfusion injury of the kidney. METHODS: Erythropoietin was injected in the peritoneal space of ICR mice after ischemia/reperfusion injury and its effect was assessed by measuring blood urea nitrogen and creatinine, and by histological analysis. Phosphorylation of Akt and endothelial nitric oxide synthase was examined by western blot analysis. Endothelial nitric oxide synthase gene null mice were also used to examine the role of endothelial nitric oxide synthase in the renoprotective effect of erythropoietin. RESULTS: Erythropoietin administration significantly inhibited the increase in blood urea nitrogen and creatinine after ischemia/reperfusion injury compared with control mice. Accordingly, erythropoietin administration significantly ameliorated the histological damages, including apoptotic cell death. Erythropoietin significantly stimulated phosphorylation of Akt and endothelial nitric oxide synthase in the kidneys. When endothelial nitric oxide synthase gene null mice were subjected to ischemia/reperfusion injury, erythropoietin did not significantly suppress the increase in blood urea nitrogen or creatinine. CONCLUSIONS: Erythropoietin seems to activate the Akt/endothelial nitric oxide synthase-dependent pathway in the kidneys. This pathway might be implicated in the renoprotective effect of erythropoietin in the ischemia/reperfusion injury model.

Development of nephrotic syndrome after administration of sorafenib in a case of metastatic renal cell carcinoma.

Nephrotic syndrome, after administration of tyrosine kinase inhibitors, is uncommon and not well known. A 62-year-old male, who had experienced a left nephrectomy due to a traffic accident 38 years ago, underwent a partial nephrectomy for right renal cell carcinoma (RCC). Histologically, the tumor was a clear cell RCC. Two years later abdominal CT revealed para-aortic lymph node metastasis. During these two years, serum creatinine had increased from 2.0 mg/dL to 2.9 mg/dL along with the appearance of proteinuria. After only a week of sorafenib, 400 mg/day, fever developed and sorafenib was stopped. Although normotensive, his serum creatinine increased to 3.83 mg/dL and serum albumin decreased from 1.8 g/L to 1.0 g/L. Proteinuria also worsened to 27.5 g/day. He became edematous, and ascites and cardiac effusions also appeared. He was diagnosed with nephrotic syndrome. A retrospective review of the histology of the partial nephrectomy revealed no change in the glomeruli.

Core lower urinary tract symptom score (CLSS) for the assessment of female lower urinary tract symptoms: a comparative study.

OBJECTIVE: We have recently developed the core lower urinary tract symptom score (CLSS) questionnaire to readily address 10 important lower urinary tract symptoms (LUTS). The aim of the present study was to evaluate the performance of the CLSS in women compared with the International Prostate Symptom Score (IPSS) and Overactive Bladder Symptom Score (OABSS). METHODS: Three hundred and eighteen treatment-naïve consecutive female patients, including 48 controls, completed the three questionnaires. Quality of life (QOL) was determined as per the IPSS QOL Index. The clinical diagnoses were overactive bladder (n = 69), mixed incontinence (n = 42), stress incontinence (n = 17), pelvic organ prolapse (n = 56), interstitial cystitis (n = 31), bacterial cystitis (n = 16), underactive bladder (n = 16), and "other" (n = 23). Simple statistics and the relationship between symptom scores and poor QOL (QOL Index ≥ 4) were examined. RESULTS: All symptom scores were significantly increased in symptomatic women. The CLSS described the symptom profiles of patients with distinct conditions. The scores of corresponding symptoms on the three questionnaires were significantly correlated (r = 0.51-0.85; all P < 0.0001). Multivariate logistic regression modeling proved five CLSS symptoms (daytime frequency, nocturia, urgency incontinence, straining, and urethral pain) as independent predictors of poor QOL, with hazard ratios ranging from 2.0 to 4.2. The IPSS included only two (urgency and straining) significant symptoms. CONCLUSIONS: The IPSS alone does not fully evaluate female LUTS, with a possible negative impact on QOL. Using the CLSS questionnaire would enable a simple and comprehensive assessment of female LUTS.