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Abstract Background Malignant gliomas exhibit rapid tumor progression and resistance to treatment, leading to high lethality. One of the causes is the reduced progression of apoptosis in glioma cells. Layilin is a type 1 transmembrane protein with a C-type lectin motif in its extracellular domain. We previously reported that layilin is mainly localized to mitochondria or their close proximity and that layilin is essential for maintaining of the fragmented type of mitochondria. This study investigates the effects of layilin on mitochondria-mediated apoptosis, focusing on B cell lymphoma (BCL)-2 family proteins in a glioma cell line of A172 cells. Results We compared the levels of pro-apoptotic BCL-2 family proteins of BAD, BAK, BAX, and BIM and anti-apoptotic BCL-2 family proteins of BCL-2 and BCL-XL between layilin- knockdown (KD) cells and control cells using western blot. The protein levels of BAD were significantly smaller in layilin-KD cells than in control cells, while those of BCL-2 were significantly larger. We then compared the mitochondrial membrane potential (ΔΨm) under p-trifluoromethoxyphenyl hydrazone (FCCP)-treated conditions using MT-1 staining. In layilin-KD cells, ΔΨm was significantly larger and FCCP-induced ΔΨm reduction was significantly lower than in control cells. Furthermore, we examined the levels of cell membrane-bound Annexin V and DNA-bound propidium idodide (PI) in layilin-KD cells with/without staurosporine (STS) treatment. Layilin-KD significantly decreased levels of cell membrane-bound Annexin V with/without STS treatment. On the other hand, PI levels were not changed by layilin-KD. We also investigated the amounts of the active caspase (CASP)-3, CASP-6, CASP-7, and poly (ADP-ribose) polymerase-1 (PARP1, cleaved form), as well as DNA fragmentation in layilin-KD cells under apoptotic conditions induced by STS, using western blot and the DNA ladder method, respectively. Under STS-treated conditions, the amounts of active CASP-3, CASP-7, and poly (ADP-ribose) PARP1 were significantly smaller in layilin-KD cells than in control cells. Accordingly, DNA fragmentation was significantly suppressed in layilin-KD cells compared to control cells under STS-treated conditions. Conclusion This study demonstrates that layilin contributes to ΔΨm reduction to promote apoptosis by up-regulating BAD and down-regulating BCL-2 in glioma cells. Our data elucidates a new function of layilin: regulation of mitochondria-mediated apoptosis.

Abstract Background Although quality of life (QOL) is an outcome of postoperative cardiac rehabilitation (CR), its course and related factors from postoperative hospitalization to the post-discharge period have not been adequately investigated. Additionally, the EuroQol-5Dimension-5Level (EQ-5D-5L) index score has not been characterized over the same period. We aimed to characterize QOL changes assessed by the EQ-5D-5L, over the period from hospitalization to 1 year post-discharge, in patients post-cardiac and thoracic aortic surgery, and investigate the factors associated with these temporal changes. Methodology This prospective, single-center study included 117 patients who underwent open cardiovascular surgery (median age, 72 years; men, 69%). Patients were assessed for QOL status when transferred to the general ward; at discharge; and at 6 and 12 months after discharge, using the EQ-5D-5L index score and a generalized linear mixed model with random intercepts. Patients were classified into two groups based on score changes post-discharge. Logistic regression analysis evaluated factors associated with QOL decrease post-discharge. Results The EQ-5D-5L index score significantly increased over time, except between 6 and 12 months post-discharge; “Common activities” was the most common dimension showing score improvement. In 25 patients (21%), the EQ-5D-5L index scores were lower after discharge compared to their scores at discharge. In the logistic regression analysis, Barthel Index pre-admission, preoperative hemoglobin level, and Mini-Mental State Examination-Japanese scores pre-discharge were significantly associated with QOL decline after adjusting for the European System for Cardiac Operative Risk Evaluation II score. Conclusions Most patients post-cardiac or thoracic aortic surgery experienced improved QOL from postoperative hospital stay to 1 year post-discharge. However, in patients with pre-operative basic activities of daily living, hemoglobin and post-operative cognitive decline may require ongoing comprehensive CR because of reduced QOL. Given the potential selection bias introduced by the relatively small sample size in this study, future research involving larger populations is necessary.

Abstract Background An arteriovenous fistula (AVF) is the most common type of vascular access for hemodialysis. As it causes volume overload and sometimes increases pulmonary artery pressure, it is unsuitable for some patients. Herein, we describe a patient with acute kidney disease who required maintenance hemodialysis with vascular access other than an AVF owing to post-lung transplant pulmonary hypertension. Case presentation A 50-year-old man with interstitial pneumonia underwent living-donor lobar lung transplantation at our hospital. Weaning from venoarterial extracorporeal membrane oxygenation was achieved; however, the patient required mechanical ventilation owing to pulmonary hypertension. He developed acute kidney disease and required maintenance hemodialysis with sustainable vascular access. Although echocardiography showed a normal ejection fraction, we expected volume overload after arteriovenous access construction to worsen his pulmonary hypertension because of his inadequate pulmonary vascular bed. Therefore, a tunneled central vein catheter was implanted into the right femoral vein as a bridge, and superficialization of the right brachial artery was performed for long-term vascular access. Conclusions As this patient had sustained post-transplant pulmonary hypertension and small grafts, we avoided creating arteriovenous access because of concern over the aggravation of pulmonary hypertension. Evaluation of right heart function and pulmonary hypertension is important before arteriovenous access construction.

Macrophages contribute to post-transplant lung rejection. Disulfiram (DSF), an anti-alcoholic drug, has an anti-inflammatory effect and regulates macrophage chemotactic activity. Here, we investigated DSF efficacy in suppressing acute rejection post-lung transplantation. Male Lewis rats (280–300 g) received orthotopic left lung transplants from Fisher 344 rats (minor histocompatibility antigen-mismatched transplantation). DSF (0.75 mg/h) monotherapy or co-solvent only (50% hydroxypropyl-β-cyclodextrin) as control was subcutaneously administered for 7 days (n = 10/group). No post-transplant immunosuppressant was administered. Grades of acute rejection, infiltration of immune cells positive for CD68, CD3, or CD79a, and gene expression of monocyte chemoattractant protein and pro-inflammatory cytokines in the grafts were assessed 7 days post-transplantation. The DSF-treated group had significantly milder lymphocytic bronchiolitis than the control group. The infiltration levels of CD68+ or CD3+ cells to the peribronchial area were significantly lower in the DSF than in the control groups. The normalized expression of chemokine ligand 2 and interleukin-6 mRNA in allografts was lower in the DSF than in the control groups. Validation assay revealed interleukin-6 expression to be significantly lower in the DSF than in the control groups. DSF can alleviate acute rejection post-lung transplantation by reducing macrophage accumulation around peripheral bronchi and suppressing pro-inflammatory cytokine expression.

Abstract Truncation of the C‐terminal tail of the β2‐AR, transfection of βARKct or over‐expression of a kinase‐dead GRK mutant reduces isoprenaline‐stimulated glucose uptake, indicating that GRK is important for this response. We explored whether phosphorylation of the β2‐AR by GRK2 has a role in glucose uptake or if this response is related to the role of GRK2 as a scaffolding protein. CHO‐GLUT4myc cells expressing wild‐type and mutant β2‐ARs were generated and receptor affinity for [3H]‐CGP12177A and density of binding sites determined together with the affinity of isoprenaline and BRL37344. Following receptor activation by β2‐AR agonists, cAMP accumulation, GLUT4 translocation, [3H]‐2‐deoxyglucose uptake, and β2‐AR internalization were measured. Bioluminescence resonance energy transfer was used to investigate interactions between β2‐AR and β‐arrestin2 or between β2‐AR and GRK2. Glucose uptake after siRNA knockdown or GRK inhibitors was measured in response to β2‐AR agonists. BRL37344 was a poor partial agonist for cAMP generation but displayed similar potency and efficacy to isoprenaline for glucose uptake and GLUT4 translocation. These responses to β2‐AR agonists occurred in CHO‐GLUT4myc cells expressing β2‐ARs lacking GRK or GRK/PKA phosphorylation sites as well as in cells expressing the wild‐type β2‐AR. However, β2‐ARs lacking phosphorylation sites failed to recruit β‐arrestin2 and did not internalize. GRK2 knock‐down or GRK2 inhibitors decreased isoprenaline‐stimulated glucose uptake in rat L6 skeletal muscle cells. Thus, GRK phosphorylation of the β2‐AR is not associated with isoprenaline‐ or BRL37344‐stimulated glucose uptake. However, GRKs acting as scaffold proteins are important for glucose uptake as GRK2 knock‐down or GRK2 inhibition reduces isoprenaline‐stimulated glucose uptake.

Living-donor lobar lung transplantation (LDLLT) is a well-established surgical procedure with favorable outcomes; however, the frequency of readmission and costs in LDLLT are poorly understood. Here, we aimed to compare health care costs and readmissions at 90 days and 1 year after the index discharge in LDLLT and cadaveric lung transplantation (CLT) and evaluate the reasons for readmission and in-hospital mortality. In this retrospective cohort study, we used the Diagnosis Procedure Combination database, a nationwide inpatient database in Japan to obtain initial lung transplantation data for all patients from July 2010 to March 2020. Multivariable Poisson or multiple regression analyses after multiple imputation was performed to compare the cumulative number of readmissions and costs between patients receiving LDLLT and CLT. Among 514 recipients, 115 (22%) underwent LDLLT and 399 (78%) received CLT. Overall, in-hospital mortality after transplantation was 4.5%. The LDLLT group showed a significantly lower crude readmission rate (90 days, 22% vs 37%, p = 0.004; 1 year, 48% vs 62%, p = 0.031) and median readmission cost (90 days, United States dollar (USD) 0 vs 0, p = 0.003; 1 year, USD 1178 vs 4714, p = 0.005) than the CLT group. Multivariable regression analyses showed that the LDLLT group had a lower risk of readmission (incidence rate ratio, 0.59; 95% confidence interval, 0.38-0.92; p = 0.020) and lower costs at 90 days (difference, USD −11,629; 95% confidence interval, −5682 to −17,462; p

Abstract Lung transplantation (LT) is the only option for patients with pulmonary arterial hypertension (PAH) refractory to maximal medical therapy. However, some patients referred for LT could survive without LT, and its determinants remain unclear. This study aimed to elucidate prognostic factors of severe PAH at the referral time. We retrospectively analyzed 34 patients referred for LT evaluation. The primary outcome was a composite of death or LT. Over a median follow-up period of 2.56 years, eight patients received LT and eight died. Compared with LT-free survival group, pulmonary arterial systolic pressure (PASP) was higher (p = 0.042), and the ratio of tricuspid annular plane systolic excursion (TAPSE) to PASP (TAPSE/PASP) was lower (p = 0.01) in LT or death group. In receiver operating characteristic analysis, the area under the curve was 0.759 (95% confidence interval 0.589–0.929) for TAPSE/PASP to predict primary outcome, and the optimal cut-off value was 0.30 mm/mmHg (sensitivity 0.875 and specificity 0.667). In a multivariate analysis, TAPSE/PASP was independently associated with death or LT. Kaplan–Meier analysis showed a better LT-free survival in patients with TAPSE/PASP ≧0.30 mm/mmHg than in those with

Monitoring of artificial reefs (ARs) has been conducted through such methods as visual censuses, surveys using fishing gear, and echo sounder. These methods have disadvantages: visual census is not possible at ARs in deeper waters, fishing gear surveys are invasive to fish individuals, and echo sounders have difficulty in species identification. A new AR monitoring method is required to compensate for these disadvantages. While eDNA has become a valid monitoring tool for marine biodiversities, it is influenced by degradation and transport of the molecules that affect information about the spatio-temporal distribution of fish. An understanding of the relationship between current fields and eDNA distribution, particularly in open waters, is critical when using eDNA as an index for fish aggregation at ARs. We investigated the relationship between eDNA distribution and current fields around an AR for four dominant species (Engraulis japonicus, Parapristipoma trilineatum, Scomber spp and Trachurus japonicus) in Tateyama Bay, Japan. The highest density of fish schools is formed directly above or at the upstream side of ARs. If we assume that the center of eDNA originates at these locations at an AR and eDNA is simply transported by currents, a higher density of eDNA would distribute downstream from the AR. However, our results indicate that eDNA distribution is in accord with actual fish distribution, namely eDNA densities are more abundant in the upstream side of ARs. We thus consider that eDNA distribution is more influenced by actual distribution patterns than by the transport processes.

Abstract Background Thrombotic microangiopathy is a syndrome characterized by microangiopathic hemolytic anemia and platelet aggregation, which is caused by endothelial injury, microcirculation thrombosis, and fibrin deposition. Transplant-associated thrombotic microangiopathy rarely occurs after lung transplantation and the onset is generally later than that after bone marrow or other solid organ transplantation. The treatment is to stop administration of the causal agent, which is often a calcineurin inhibitor, such as tacrolimus and cyclosporine. We herein report the case of a patient with early post-transplant thrombotic microangiopathy after lung transplantation treated by introducing basiliximab and temporarily stopping any calcineurin inhibitors until resuming treatment with an alternative calcineurin inhibitor. Case presentation A 58-year-old Asian woman underwent bilateral lung transplantation for hypersensitivity pneumonitis caused by an avian antigen, or bird fancier’s lung disease. Postoperatively, she was started on triple immunosuppressive therapy, which included tacrolimus, mycophenolate mofetil, and steroids. On postoperative day 6, she developed thrombocytopenia followed by fever, hemolytic anemia, renal dysfunction, and purpura on her limbs and abdomen. She was diagnosed with transplant-associated thrombotic microangiopathy, and tacrolimus was thought to be the causal agent. We stopped tacrolimus and administered basiliximab. Then, she developed oliguria and needed continuous hemodiafiltration. On postoperative day 14, the platelet count recovered and she was switched from basiliximab to cyclosporine. Using this protocol, worsening thrombotic microangiopathy and acute rejection were avoided. Conclusions We report the case of a patient with early post-transplant thrombotic microangiopathy after lung transplantation that was treated with basiliximab. Switching from calcineurin inhibitors using basiliximab may be an option for treating thrombotic microangiopathy without increasing the risk of acute rejection.

Abstract Background Thymic epithelial tumors (TETs) exhibit irregular shapes reflective of the heterogeneity in tumor growth and invasive properties. We aimed to identify the prognostic value of the pathological tumor long‐to‐short axis (L/S) ratio in TETs. Methods A retrospective study was performed on patients with TETs who underwent extended thymectomy between January 1999 and December 2019 in our institute. Patients were divided into two groups according to the threshold of the L/S ratio. Overall survival (OS) and progression‐free survival (PFS) were evaluated by Kaplan‐Meier analysis. The independent prognostic factors of TETs were identified by multivariate analysis. The performance of prediction models for the above survival outcomes with and without the L/S ratio was evaluated using an integrated time‐dependent area under the curve (iAUC). Results Eligible patients were divided into two groups based on higher (n = 42) and lower (n = 94) L/S ratios according to a threshold value of 1.39. A significant difference was found between the two groups only in disease progression (p = 0.001). Poorer survival outcomes were found from Kaplan‐Meier curves in the higher L/S ratio group (p 1.39 is associated with worse survival outcomes.

Abstract Rapid identification of lung-cancer micro-lesions is becoming increasingly important to improve the outcome of surgery by accurately defining the tumor/normal tissue margins and detecting tiny tumors, especially for patients with low lung function and early-stage cancer. The purpose of this study is to select and validate the best red fluorescent probe for rapid diagnosis of lung cancer by screening a library of 400 red fluorescent probes based on 2-methyl silicon rhodamine (2MeSiR) as the fluorescent scaffold, as well as to identify the target enzymes that activate the selected probe, and to confirm their expression in cancer cells. The selected probe, glutamine-alanine-2-methyl silicon rhodamine (QA-2MeSiR), showed 96.3% sensitivity and 85.2% specificity for visualization of lung cancer in surgically resected specimens within 10 min. In order to further reduce the background fluorescence while retaining the same side-chain structure, we modified QA-2MeSiR to obtain glutamine-alanine-2-methoxy silicon rhodamine (QA-2OMeSiR). This probe rapidly visualized even borderline lesions. Dipeptidyl peptidase 4 and puromycin-sensitive aminopeptidase were identified as enzymes mediating the cleavage and consequent fluorescence activation of QA-2OMeSiR, and it was confirmed that both enzymes are expressed in lung cancer. QA-2OMeSiR is a promising candidate for clinical application.

Abstract Major psychiatric disorders such as attention-deficit/hyperactivity disorder and schizophrenia are often accompanied by elevated impulsivity. However, anti-impulsive drug treatments are still limited. To explore a novel molecular target, we examined the role of dopamine D5 receptors in impulse control using mice that completely lack D5 receptors (D5KO mice). We also measured spontaneous activity and learning/memory ability because these deficits could confound the assessment of impulsivity. We found small but significant effects of D5 receptor knockout on home cage activity only at specific times of the day. In addition, an analysis using the q-learning model revealed that D5KO mice displayed lower behavioral adjustment after impulsive actions. However, our results also showed that baseline impulsive actions and the effects of an anti-impulsive drug in D5KO mice were comparable to those in wild-type littermates. Moreover, unlike previous studies that used other D5 receptor-deficient mouse lines, we did not observe reductions in locomotor activity, working memory deficits, or severe learning deficits in our line of D5KO mice. These findings demonstrate that D5 receptors are dispensable for impulse control. Our results also indicate that time series analysis and detailed analysis of the learning process are necessary to clarify the behavioral functions of D5 receptors.

Abstract Previous findings have proposed that drugs targeting 5-HT2C receptors could be promising candidates in the treatment of trauma- and stress-related disorders. However, the reduction of conditioned freezing observed in 5-HT2C receptor knock-out (KO) mice in previous studies could alternatively be accounted for by increased locomotor activity. To neutralize the confound of individual differences in locomotor activity, we measured a ratio of fear responses during versus before the presentation of a conditioned stimulus previously paired with a footshock (as a fear measure) by utilizing a conditioned licking suppression paradigm. We first confirmed that 5-HT2C receptor gene KO attenuated fear responses to distinct types of single conditioned stimuli (context or tone) independently of locomotor activity. We then assessed the effects of 5-HT2C receptor gene KO on compound fear responses by examining mice that were jointly conditioned to a context and a tone and later re-exposed separately to each. We found that separate re-exposure to individual components of a complex fear memory (i.e., context and tone) failed to elicit contextual fear extinction in both 5-HT2C receptor gene KO and wild-type mice, and also abolished differences between genotypes in tone-cued fear extinction. This study delineates a previously overlooked role of 5-HT2C receptors in conditioned fear responses, and invites caution in the future assessment of molecular targets and candidate therapies for the treatment of PTSD.

A 16-year-old girl with a genetic diagnosis of cystic fibrosis was referred to us for consideration of lung transplantation. She had been hospitalized repeatedly for pneumonia and pneumothoraxes and her respiratory function had worsened progressively. Although she also had liver cirrhosis, she was considered a candidate for lung transplantation because her liver disease was compensated and only slowly progressive. After bilateral lung transplantation from a brain-dead donor, she developed ascites that was well controlled with diuretics. Otherwise, her post-operative course was uneventful and she was transferred to another hospital for rehabilitation 39 days after lung transplantation.

A 45-year-old woman with recurrent dyspnea for 40 years was previously diagnosed with bronchial asthma and spasmodic dysphonia. On admission, the patient was diagnosed with expiratory central airway collapse (ECAC) due to expiratory dynamic airway collapse based on radiographic examination, chest computed tomography, and bronchoscopy. After continuous positive airway pressure and temporal airway stenting, surgical tracheobronchoplasty and tracheal membranous portion reinforcement using polypropylene mesh successfully relieved the respiratory symptoms. In patients with airway obstructive disease refractory to conventional therapies, ECAC should be considered.

Introduction: Lung cancer presenting as ground-glass nodules (GGNs) with little consolidation is often diagnosed as noninvasive carcinoma. Increasing evidence suggests that sublobar lung resection, such as wedge resection, provides satisfactory long-term outcomes equivalent to those of lobectomy. Spread through alveolar spaces (STAS) is a reported risk factor for recurrence, especially when sublobar lung resection is applied. We herein report a rare case of a pure GGN that was histologically confirmed to be adenocarcinoma in situ (AIS) and associated STAS after limited resection. Case presentation: A 67-year-old woman presented with an 8-mm pure GGN in S2 of the right upper lobe and a part-solid GGN in S8 of the right lower lobe. The clinical stages of these GGNs were TisN0M0 stage 0 and T1aN0M0 stage IA1, respectively. The patient underwent video-assisted thoracoscopic segmentectomies of S2 and S8. Histopathological examination of the resected specimens revealed minimally invasive adenocarcinoma in S8 and AIS in S2. Stromal invasion was not observed in the tumor located in S2, but a tumor cell cluster was found in the air space located away from the AIS lesion. Thus, the lesion was positive for STAS. Conclusion: We have reported a rare case of a pure GGN that was histologically STAS-positive, otherwise diagnosed as AIS after limited resection.

Abstract Since the early 1970s, many artificial reefs (ARs) have been deployed in Japanese coastal waters to create fisheries grounds. Recently, researchers began to use environmental DNA (eDNA) methods for biodiversity monitoring of aquatic species. A metabarcoding approach using internal standard DNAs [i.e., quantitative MiSeq sequencing (qMiSeq)] makes it possible to monitor eDNA concentrations of multiple species simultaneously. This method can improve the efficiency of monitoring AR effects on fishes. Our study investigated distributions of marine fishes at ARs and surrounding stations in the open oceanographic environment of Tateyama Bay, central Japan, using qMiSeq and echo sounder survey. Using the qMiSeq with 12S primers, we found higher quantities of fish eDNAs at the ARs than at surrounding stations and different fish species compositions between them. Comparisons with echo sounder survey also showed positive correlations between fish eDNA concentration and echo intensity, which indicated a highly localized signal of eDNA at each sampling station. These results suggest that qMiSeq is a promising technique to complement conventional methods to monitor distributions of multiple fish species.

Abstract Background Although the number of patients who undergo extracorporeal membrane oxygenation (ECMO) as a bridge to lung transplantation is increasing worldwide, there are few reports on lung transplantation after long-term ECMO (more than 1 month). We report a rare case of successful bilateral lung transplantation in a Japanese patient after 5 months of veno-venous (VV)-ECMO support. Case presentation A 27-year-old man who underwent bone marrow transplantation (BMTx) with fully matched human leukocyte antigen typing was diagnosed with bronchiolitis obliterans caused by chronic graft-versus-host disease 3 years after the BMTx. One year later, his respiratory condition had exacerbated, with carbon dioxide retention that required conventional mechanical ventilation. He was then deemed a suitable candidate for lung transplantation by a multidisciplinary transplantation selection committee. While waiting for donor lungs, his hypercapnia and acidosis became barely manageable under care with mechanical ventilation and ultimately he was switched to VV-ECMO. He remained on VV-ECMO for the next 5 months, during which period the circuit was switched nine times. In addition, sophisticated intensive care was required to manage multiple episodes of sepsis and coagulopathy. A suitable donor was identified 5 months later, and bilateral lung transplantation was initiated with continuous VV-ECMO. The procedure itself was extremely challenging owing to severe adhesions resulting from previous thoracotomy, inflammation, infection, and intrapulmonary hemorrhage. The operative time for the transplantation was about 19 h. Currently, at 2 years 8 months postoperatively, the patient is alive and well. Conclusion Transplant surgery in this patient was extremely challenging because of the presence of severe pleural adhesions and stiff native lungs secondary to hemorrhagic complications due to the prolonged ECMO support. Surgeons must recognize that lung transplantation after long-term ECMO bridging can be technically more complicated and challenging than shorter-term ECMO.

Abstract Background A bronchial fistula is a relatively rare and potentially fatal complication after lung transplantation. Thoracic surgeons and pulmonologists often face challenges when selecting treatment options. We herein report an exceptional case of intrabronchial migration of a nonabsorbable hemostatic agent, which had been placed around the pulmonary artery at the time of lung transplantation, through a bronchial fistula. Case presentation A 61-year-old man developed respiratory distress 1 year after left single-lung transplantation for idiopathic interstitial pneumonia. Bronchoscopic examination revealed an apparent foreign body protruding from the mediastinum into the distal site of the bronchial anastomosis. The foreign body was easily removed bronchoscopically and appeared to be a hemostatic agent that had been placed during the previous lung transplantation. The patient developed a similar clinical episode and finally developed hemoptysis. Computed tomography revealed a foreign body located between the bronchus and pulmonary artery, partially protruding into the bronchial lumen. Given the possibility of a bronchopulmonary arterial fistula, surgical treatment was performed. The foreign body was located between the bronchus and left pulmonary artery and was easily removed. Multiple bronchial fistulas were found, and all were closed with direct sutures. Bypass grafting of the left pulmonary artery was then performed, initially with a homograft but eventually with an extended polytetrafluoroethylene graft. The patient was finally discharged 5 months after the surgery. Conclusion We experienced an extremely rare case of intrabronchial migration of hemostatic agents used during the previous lung transplantation through a bronchial fistula, which were successfully managed by direct bronchial closure and bypass grafting of the left pulmonary artery.

Abstract Background Bronchial necrosis is a rare but fatal complication after radiation therapy. Because of the anatomical complexity and rarity of this condition, determining the most appropriate management for individual patients is extremely challenging. Lung autotransplantation is a surgical technique that has been applied to hilar neoplastic lesions to preserve pulmonary function and avoid pneumonectomy. We herein report a case of bronchial necrosis secondary to radiotherapy that was treated with lung autotransplantation. Case presentation A 46-year-old man developed broad necrosis and infection of the right bronchus secondary to previous stereotactic body-radiation therapy. This treatment was supplied close to a right hilar metastatic pulmonary tumor derived from a mediastinal malignant germ cell tumor that had been surgically resected with the left phrenic nerve. The bronchial necrosis accompanied by infection with Aspergillus fumigatus was progressive despite antibiotics and repetitive bronchoscopic debridement. Because of the patient’s critical condition and limited pulmonary function, right lung autotransplantation with preservation of the right basal segment was selected. An omental flap was placed around the bronchial anastomosis to prevent later complications. The postoperative course involved multiple complications including contralateral pneumonia and delayed wound healing at the bronchial anastomosis with resultant stenosis, the latter of which was overcome by placement of a silicone stent. The patient was discharged 5 months postoperatively. Three months after discharge, however, the patient developed hemoptysis and died of bronchopulmonary arterial fistula formation. Conclusions We experienced an extremely challenging case of bronchial necrosis secondary to radiotherapy. The condition was managed with lung autotransplantation and omental wrapping; however, the treatment success was temporary and the patient eventually died of bronchopulmonary arterial fistula formation. This technique seems to be a feasible option for locally advanced refractory bronchial necrosis, although later complications can still be fatal.

Mirror therapy (MT) helps stroke survivors recover motor function. Previous studies have reported that an individual’s motor imagery ability is related to the areas of brain activity during motor imagery and the effectiveness of motor imagery training. However, the relationship between MT and motor imagery ability and between corticospinal tract excitability during mirror gazing, an important component of MT, and motor imagery ability is unclear. This study determined whether the motor-evoked potential (MEP) amplitude while gazing at the mirror relates to participants’ motor imagery abilities. Twenty-four healthy right-handed adults (seven males) were recruited. Transcranial magnetic stimulation was performed while gazing at the mirror, and MEP of the first dorsal interosseous muscle of the right hand were measured. Motor imagery ability was measured using the Kinesthetic and Visual Imagery Questionnaire (KVIQ), which assesses the vividness of motor imagery ability. Additionally, a mental chronometry (MC) task was used to assess time aspects. The results showed a significant moderate correlation between changes in MEP amplitude values while gazing at the mirror, as compared with resting conditions, and assessment scores of KVIQ. This study shows that corticospinal excitability because of mirror gazing may be related to the vividness of motor imagery ability.

BackgroundThe number of mobile health (mHealth) apps continues to rise each year. Widespread use of the Mobile App Rating Scale (MARS) has allowed objective and multidimensional evaluation of the quality of these apps. However, no Japanese version of MARS has been made available to date. ObjectiveThe purposes of this study were (1) to develop a Japanese version of MARS and (2) to assess the translated version’s reliability and validity in evaluating mHealth apps. MethodsTo develop the Japanese version of MARS, cross-cultural adaptation was used using a universalist approach. A total of 50 mental health apps were evaluated by 2 independent raters. Internal consistency and interrater reliability were then calculated. Convergent and divergent validity were assessed using multitrait scaling analysis and concurrent validity. ResultsAfter cross-cultural adaptation, all 23 items from the original MARS were included in the Japanese version. Following translation, back-translation, and review by the author of the original MARS, a Japanese version of MARS was finalized. Internal consistency was acceptable by all subscales of objective and subjective quality (Cronbach α=.78-.89). Interrater reliability was deemed acceptable, with the intraclass correlation coefficient (ICC) ranging from 0.61 to 0.79 for all subscales, except for “functionality,” which had an ICC of 0.40. Convergent/divergent validity and concurrent validity were also considered acceptable. The rate of missing responses was high in several items in the “information” subscale. ConclusionsA Japanese version of MARS was developed and shown to be reliable and valid to a degree that was comparable to the original MARS. This Japanese version of MARS can be used as a standard to evaluate the quality and credibility of mHealth apps.

Abstract Background Systemic scleroderma (SSc) involves multiple organs including the skin, the lung, the kidney, and the esophagus. Nowadays, patient life prognosis has substantially improved due to more appropriate management of lung complications, including lung transplantation. However, the extension of their survival may increase SSc patients with chronic kidney diseases and requiring renal replacement therapy (RRT). Case presentation A 51-year-old female with SSc who underwent unilateral deceased-donor lung transplantation was referred because of progressive renal dysfunction. Despite no episodes of scleroderma renal crisis, her renal function gradually deteriorated for 2 years with her serum creatinine level increasing from 0.5 mg/dL at transplantation to 4.3 mg/dL. Although we reinforced antihypertensive treatment and reduced calcineurin inhibitor dose, she thereafter developed symptomatic uremia. Due to impaired manual dexterity with contracture of the interphalangeal joints, no caregivers at home, and kidney transplantation donor unavailability, maintenance hemodialysis was chosen as RRT modality. Further, due to the narrowing of superficial vessels in the sclerotic forearm skin and post-transplant immunocompromised status, the native left brachiocephalic arteriovenous fistula was created. Post-operative course was uneventful while any sign of cutaneous infection and pulmonary hypertension was closely monitored. Our literature review also indicates several difficulties with initiating and maintaining RRT in patients with SSc although case reports of kidney failure after lung transplanation in SSc were not accumulated. Conclusions With respect to initiating RRT for post-lung transplant patients with SSc, the clinical course of our case exemplifies recent complex trends of renal management. The optimal modality with secured initiation of RRT should be carefully determined based on the severity and risk for the cardiopulmonary, peripheral vascular, cutaneous, and systemic or local infectious complications.

Abstract Background Idiopathic pleuroparenchymal fibroelastosis (IPPFE) is a rare lung disease that manifests as parenchymal fibrosis of the upper lung lobe and pleura. There have been no reports of IPPFE complicating pregnancy. Here, we report a case of IPPFE that deteriorated rapidly during pregnancy. Case presentation A 29-year-old woman presented with dyspnea and dry cough at 19 weeks of gestation. IPPFE with acute exacerbation was suspected on chest computed tomography (CT). Despite steroid treatment, her condition progressed. A cesarean section was performed at 28 weeks of gestation. On postoperative day 26, she underwent living-donor lung transplantation. She was discharged a year after transplantation. Conclusion Our experience suggested that when pregnancy is complicated by PPFE, the disease may deteriorate rapidly. In this case, even though IPPFE with acute exacerbation was diagnosed during pregnancy, live birth was achieved, and the mother survived after lung transplantation. Lung transplantation should be considered in these patients because, once advanced, pulmonary lesions may be irreversible.

The ATP1A2 coding α2 subunit of Na,K‐ATPase, which is predominantly located in astrocytes, is a causative gene of familial hemiplegic migraine type 2 (FHM2). FHM2 model mice (Atp1a2tmCKwk/+) are susceptible to cortical spreading depression (CSD), which is profoundly related to migraine aura and headache. However, astrocytic properties during CSD have not been examined in FHM2 model mice. Using Atp1a2tmCKwk/+ crossed with transgenic mice expressing G‐CaMP7 in cortical neurons and astrocytes (Atp1a2+/−), we analyzed the changes in Ca2+ concentrations during CSD. The propagation speed of Ca2+ waves and the percentages of astrocytes with elevated Ca2+ concentrations in Atp1a2+/− were higher than those in wild‐type mice. Increased percentages of astrocytes with elevated Ca2+ concentrations in Atp1a2+/− may contribute to FHM2 pathophysiology.

BackgroundEarly rehabilitation allows patients to better perform the activities of daily living after hospital discharge. A specialized physical therapist has been assigned as part of the early rehabilitation, but the effectiveness of the program remains unclear. We investigated how early rehabilitation provided by a specialized physical therapist affects ADL in patients with sepsis.MethodsThis was a retrospective cohort study. This study's subjects were sepsis patients who entered the advanced emergency critical care center of Shinshu University Hospital between April 2014 and March 2020. Electronic medical records were reviewed to obtain information on demographic characteristics, severity score, primary source of infection, therapeutic medication, the number of days after hospital admittance until rehabilitation begins, length of hospital stay, discharge to home, and an assessment of daily living activities for each patient. The patients were divided into two groups based on whether they were treated before or after a specialized physical therapist had been hired by the advanced emergency critical care center.ResultsAssigning a physical therapist to a patient significantly shortened the number of days until rehabilitation began. In a multivariable model, the strongest predictors of return to independent living after hospital discharge were (1) assigning a specialized physical therapist (odds ratio = 2.40; 95% confidence interval = 1.09-5.79; P = 0.050) and (2) the number of days until rehabilitation started (odds ratio = 0.24; 95% confidence interval = 0.08-0.76; P = 0.014).ConclusionsAssigning a specialized physical therapist to sepsis patients at an advanced emergency critical care center significantly shortened the number of days until a patient can begin rehabilitation after hospital admittance and improved activities of daily living after hospital discharge.Trial registrationTrial registration [University Hospital Medical Information Network Clinical Trials Registry, number UMIN000040570 (2020/5/28).].

Abstract Background Bovine leukemia virus (BLV), which is closely related to human T-cell leukemia virus, is the etiological agent of enzootic bovine leukosis, a disease characterized by a highly prolonged course involving persistent lymphocytosis and B-cell lymphoma. The bovine major histocompatibility complex class II region plays a key role in the subclinical progression of BLV infection. In this study, we aimed to evaluate the roles of CD4+ T-cell epitopes in disease progression in cattle. Methods We examined five Japanese Black cattle, including three disease-susceptible animals, one disease-resistant animal, and one normal animal, classified according to genotyping of bovine leukocyte antigen (BoLA)-DRB3 and BoLA-DQA1 alleles using polymerase chain reaction sequence-based typing methods. All cattle were inoculated with BLV-infected blood collected from BLV experimentally infected cattle and then subjected to CD4+ T-cell epitope mapping by cell proliferation assays. Results Five Japanese Black cattle were successfully infected with BLV, and CD4+ T-cell epitope mapping was then conducted. Disease-resistant and normal cattle showed low and moderate proviral loads and harbored six or five types of CD4+ T-cell epitopes, respectively. In contrast, the one of three disease-susceptible cattle with the highest proviral load did not harbor CD4+ T-cell epitopes, and two of three other cattle with high proviral loads each had only one epitope. Thus, the CD4+ T-cell epitope repertoire was less frequent in disease-susceptible cattle than in other cattle. Conclusion Although only a few cattle were included in this study, our results showed that CD4+ T-cell epitopes may be associated with BoLA-DRB3-DQA1 haplotypes, which conferred differential susceptibilities to BLV proviral loads. These CD4+ T-cell epitopes could be useful for the design of anti-BLV vaccines targeting disease-susceptible Japanese Black cattle. Further studies of CD4+ T-cell epitopes in other breeds and using larger numbers of cattle with differential susceptibilities are required to confirm these findings.

Peripheral sensory nerve electrical stimulation (PES) excites the primary motor cortex and is expected to improve motor dysfunction post-stroke. However, previous studies have reported a variety of stimulus frequencies and stimulus duration settings, and the effects of these different combinations on primary motor cortex excitability are not clear. We aimed to clarify the effects of different combinations of stimulus frequency and stimulus duration of PES on the excitation of primary motor cortex. Twenty-one healthy individuals (aged > 18 years, right-handed, and without a history of neurological or orthopedic disorders) were included. Each participant experienced three different stimulation frequencies (1, 10 and 50 Hz) and durations (20, 40 and 60 min). Motor-evoked potentials (MEPs) were recorded pre- and post-PES. The outcome measure was the change in primary motor cortex excitability using the MEP ratio. We used a D-optimal design of experiments and response surface analysis to define the optimal combination within nine different settings inducing more satisfying responses. The combination of stimulation frequency and stimulation time that maximized the desirability value was 10 Hz and 40 min, respectively. The results of this study may provide fundamental data for more minimally invasive and effective implementation of PES in patients with stroke.

Background The current staging systems do not consider the tumor location of thymomas, and its clinical relevance is poorly understood. This study aimed to evaluate the impact of tumor location on the clinicopathological features and prognosis of thymomas. Methods We performed a retrospective review of patients at our institution who underwent an extended thymectomy for a thymoma from 1976 to 2015. The tumor location was classified as either the superior or inferior mediastinum based on the maximum tumor diameter. The clinicopathological characteristics of the thymoma were also evaluated. Kaplan‐Meier estimates and Cox proportional hazards models were used to analyze the survival outcomes and risk factors for recurrence. Results A total of 194 patients with thymoma were eligible for this study. Compared with the inferior mediastinum group (n = 167), the superior mediastinum group (n = 27) had a higher frequency of myasthenia gravis (MG), advanced Masaoka‐Koga staging, disease progression and recurrence (P

The brain stores memories through a set of neurons known as engram cells. Here, the authors show that engram cells in the mouse hippocampus are organized into sub-ensembles representing distinct pieces of information, which are then orchestrated to constitute an entire memory.

Summary: Background/Objective: Virtual assisted lung mapping (VAL-MAP) is a bronchoscopic lung marking technique developed to assist in navigational lung resection. It can be used for nodule localization and segmental identification. This article presents our initial experience of thoracoscopic pulmonary segmentectomy using combined VAL-MAP and computed tomography (CT)-guided localization. Material and methods: Markings with India Ink were made bronchoscopically, before surgery, using a virtual bronchoscopy system (LungPoint® Planner) without fluoroscopy guidance. Post VAL-MAP CT scans localized the actual markings. All data on patients, markings, and outcomes were retrospectively collected, and the contribution of VAL-MAP to the operation was graded by the surgeon. Results: From March 2017 to September 2017, 24 consecutive patients received the VAL-MAP marking procedure before thoracoscopic segmentectomy. Nineteen patients also received pre-operative CT-guided percutaneous localization after VAL-MAP; fifteen patients received CT-guided localization with dye (patent blue V) and microcoil, and four patients received with dye only. Of the 101 marking attempts made in all the patients, 71 (70.3%) were identified as contributing to the surgery. No clinically evident complications were associated with the procedure. A total of 24 segmentectomies were thoracoscopically conducted for 18 cases of lung cancer and six cases of benign diseases. Conclusion: The combination of VAL-MAP and CT-guided percutaneous localization contribute to precise thoracoscopic pulmonary segmentectomy. Keywords: VAL-MAP, CT-Guided localization, Segmentectomy, Lung cancer

A disintegrin and metalloprotease 17 (ADAM17) catalyzes the cleavage and release of the ectodomains of its substrates at the cell surface in a process termed ectodomain shedding. However, not all ADAM17 substrates have been identified. Here, we used cell surface protein-specific labeling and proteomic approaches to detect and identify ADAM17 substrates. HeLa cell surface proteins were labeled with a fluorescent dye and cultured with or without TAPI-2, an ADAM17 inhibitor. Labeled proteins released into the culture medium were detected by 2-dimensional gel electrophoresis (2DE). Protein spots showing decreased intensity in response to TAPI-2 were selected as substrates of ADAM17 or their binding proteins, and identified by mass spectrometry. ADAM17 knockdown was preformed to examine the behavior of identified proteins. Of 347 proteins detected by 2DE, 49 showed lower intensity in TAPI-2 (+) than in TAPI-2 (-) samples (p < 0.05), and were considered as candidate substrates of ADAM17. Mass spectrometric analysis of 14 protein spots showing >50% decreased intensity identified clusterin as a novel ADAM17 substrate, in addition to known substrates such as desmoglein-2. Western blot analysis showed that ADAM17 knockdown decreased the levels of clusterin fragments cleaved and released from the cell surface. The results identified clusterin as a novel ADAM17 substrate. The method used to identify clusterin could be used to identify the substrates of other sheddases involved in ectodomain shedding.

Background: Antagonistic tasks are cognitive-motor task trainings. Intervention programs involving antagonistic exercise tasks are being employed to help prevent falls and reduce the need for nursing care in older populations. Meanwhile, the effects of such tasks on blood flow in the brain remain obscure. This study aimed to clarify the effects of antagonistic tasks on prefrontal cortical cerebral hemodynamics.Materials and Methods: We assessed 13 healthy adults (two men, 11 women; mean age, 21.4 ± 1.0 years). Participants imitated each of the antagonistic tasks presented on a PC monitor placed at a 120-mm viewing distance. All participants performed six tasks, consisting of upper-limb tasks (non-antagonism, simple antagonism, and complex antagonism) and upper- and lower-limb tasks (tasks combining lower-limb opening and closing movements with each upper-limb task). We used near-infrared spectroscopy (NIRS) to measure cerebral blood flow dynamics, with oxygenated hemoglobin (Oxy-Hb) concentration changes as the main outcome. A 10-channel probe was placed on the participants’ forehead, focusing on the prefrontal cortex. We first obtained a baseline NIRS measurement for 10 s; the participants then imitated the task presented on the PC monitor for 90 s. We measured the number of errors and the subjective difficulty of each task.Results: The increase in prefrontal cortex Oxy-Hb concentration was significantly higher in the complex antagonist conditions than in the non-antagonistic and simple antagonistic conditions. There were no significant prefrontal cortex Oxy-Hb differences between the upper limb and upper- and lower-limb conditions (increasing number of motor limbs).Conclusions: The study findings support that an increase in finger-shaped complexity has a greater effect on cerebral blood flow dynamics in the prefrontal cortex than does an increase in the number of motor limbs involved in the task.

Background Not all non-small cell lung cancer (NSCLC) patients possess drug-targetable driver mutations, and response rates to immune checkpoint blockade therapies also remain unsatisfactory. Therefore, more effective treatments are still needed. Here, we report the results of a phase 2 clinical trial of adoptive cell therapy using zoledronate-expanded autologous Vγ9Vδ2 T-cells for treatment-refractory NSCLC.Methods NSCLC patients who had undergone at least two regimens of standard chemotherapy for unresectable disease or had had at least one treatment including chemotherapy or radiation for recurrent disease after surgery were enrolled in this open-label, single-arm, multicenter, phase 2 study. After preliminary testing of Vγ9Vδ2 T-cell proliferation, autologous peripheral blood mononuclear cells were cultured with zoledronate and IL-2 to expand the Vγ9Vδ2 T-cells. Cultured cells (>1×109) were intravenously administered every 2 weeks for six injections. The primary endpoint of this study was progression-free survival (PFS), and secondary endpoints included overall survival (OS), best objective response rate (ORR), disease control rate (DCR), safety and immunomonitoring. Clinical efficacy was defined as median PFS significantly >4 months.Results Twenty-five patients (20 adenocarcinoma, 4 squamous cell carcinoma and 1 large cell carcinoma) were enrolled. Autologous Vγ9Vδ2 T-cell therapy was administered to all 25 patients, of which 16 completed the foreseen course of 6 injections of cultured cells. Median PFS was 95.0 days (95% CI 73.0 to 132.0 days); median OS was 418.0 days (179.0–479.0 days), and best overall responses were 1 partial response, 16 stable disease (SD) and 8 progressive disease. ORR and DCR were 4.0% (0.1%–20.4%) and 68.0% (46.5%–85.1%), respectively. Severe adverse events developed in nine patients, mostly associated with disease progression. In one patient, pneumonitis and inflammatory responses resulted from Vγ9Vδ2 T-cell infusions, together with the disappearance of a massive tumor.Conclusions Although autologous Vγ9Vδ2 T-cell therapy was well tolerated and may have an acceptable DCR, this trial did not meet its primary efficacy endpoint.Trial registration number UMIN000006128

The insular cortex (IC) participates in diverse complex brain functions, including social function, yet their cellular bases remain to be fully understood. Using microendoscopic calcium imaging of the agranular insular cortex (AI) in mice interacting with freely moving and restrained social targets, we identified 2 subsets of AI neurons-a larger fraction of "Social-ON" cells and a smaller fraction of "Social-OFF" cells-that change their activity in opposite directions during social exploration. Social-ON cells included those that represented social investigation independent of location and consisted of multiple subsets, each of which was preferentially active during exploration under a particular behavioral state or with a particular target of physical contact. These results uncover a previously unknown function of AI neurons that may act to monitor the ongoing status of social exploration while an animal interacts with unfamiliar conspecifics.

Abstract Background Thymic large cell neuroendocrine carcinoma (LCNEC) is extremely rare. The detailed clinical features of thymic LNCECs remain unknown. Case presentation A 90-year-old man with a history of diabetes mellitus, chronic renal failure, and an abdominal aortic aneurysm underwent computed tomography for follow-up, which showed an anterior mediastinal tumor, measuring 31 mm × 28 mm in diameter. Magnetic resonance imaging showed an iso-intensity mass on T1-weighted images and high intensity on T2-weighted images. 18F-Fluorodeoxyglucose-positron emission tomography showed marked uptake in the mass, which was diagnosed as invasive thymoma or thymic carcinoma. Video-assisted thoracic surgery through the left thoracic cavity was converted to median sternotomy due to severe adhesions between the left lung and the chest wall. Partial thymectomy and combined partial resection of left upper lobectomy and the first and the second costal cartilages were performed. The pathologic diagnosis was thymic LCNEC, Masaoka stage III. The patient developed pleural dissemination and left lung metastases in 5 months and died 12 months after surgery. Conclusions Thymic LCNEC has high malignant potential. More cases need to be studied.

Abstract Background To understand the roles of serum exosomes in rheumatoid arthritis (RA), we comprehensively investigated the protein profiles of serum exosomes in patients with RA. Methods Exosomes were isolated from serum samples obtained from 33 patients (12 with active RA [aRA], 11 with inactive RA [iRA], 10 with osteoarthritis [OA]) and 10 healthy donors (HLs). Proteins extracted from the exosomes were separated by two-dimensional differential gel electrophoresis (2D-DIGE) and identified by mass spectrometry. Results In total, 204 protein spots were detected by 2D-DIGE. In the aRA, iRA, and OA groups, 24, 5, and 7 spots showed approximately ≥ ±1.3-fold intensity differences compared with the HL group, respectively. We were able to identify proteins in six protein spots. Among them, the protein spot identified as Toll-like receptor 3 (TLR3) showed approximately 6-fold higher intensity in the aRA group than in the other groups. Conclusions Patients with active RA possessed considerably different protein profiles of serum exosomes from patients with iRA, patients with OA, and healthy donors. The unique protein profile of serum exosomes, such as the possession of abundant TLR3 fragments, may reflect the pathophysiology of active RA.

Abstract Background Adenocarcinoma with lepidic growth pattern presents as a ground glass nodule (GGN) on high resolution computed tomography (CT), whereas peripheral pulmonary squamous cell carcinoma (SCC) usually presents as a solid nodule. We herein report a rare case of pulmonary SCC extending along the alveolar lumen representing as a GGN on a CT scan in a patient with pneumoconiosis. Case presentation A 77-year-old man with pneumoconiosis was found to have a gradually enlarging GGN in the right lower lobe of the lung on CT. An adenocarcinoma of the lung was suspected. The GGN was successfully resected by thoracoscopic segmentectomy. Pathological examination of the resected specimen was pathologically diagnosed as a stage IA SCC extending along the alveolar lumen. The patient had no evidence of recurrence 19 months after surgery. Conclusions SCC should be included in the differential diagnosis of peripherally located GGNs, especially in patients at high risk of SCC of the lung such as those with pneumoconiosis.

Abstract Background Virtual-assisted lung mapping (VAL-MAP) has been used not only in wedge resection but also in segmentectomy for hardly palpable lung nodules. We herein report a case of bilateral segmentectomy using VAL-MAP with chronological change of pulmonary function test results. Case presentation A 50-year-old female was found to have a colorectal cancer with pulmonary nodules in both sides of the lungs considered as synchronous lung metastases. After sigmoidectomy for primary cancer and chemotherapy, treatments for small nodules in both sides of the lungs were planned. Most nodules were small and supposed to be impalpable. We performed thoracoscopic segmentectomy of right S8 with the aid of VAL-MAP and, after 2 months, combined subsegmentectomy of left S8a and 9a and wide wedge resection of left S8b with the aid of VAL-MAP. All nodules suspected of lung metastases were successfully resected with adequate margins, and the decrease in pulmonary function was minimal compared with predicted postoperative forced vital capacity (FVC) and forced expiratory volume (FEV) 1.0 calculated by the numbers of subsegments. Conclusions Bilateral segmentectomies of small impalpable metastatic tumors were performed successfully with the aid of VAL-MAP.

Abstract Recently, sarcopenia has attracted attention as therapeutic target because it constitutes a risk factor for metabolic and cardiovascular diseases. We focused 5-aminolevulinic acid (ALA) which act as electron carriers in the mitochondrial electron transport system. The mice that received ALA for 8 weeks gained muscle strength and endurance, and exhibited increased muscle mass and mitochondrial amount. Administration of ALA to sarcopenia mice aged 100 weeks and chronic kidney disease (CKD) model mice also increased muscle mass and improved physical performance. Metabolome analysis revealed increased branched-chain amino acids (BCAAs) levels in the skeletal muscle of ALA-treated mice. Quantitative PCR analysis revealed decreased expression levels in branched-chain amino acid transaminases (BCATs) that degrade BCAAs and other muscle-degrading factors, and increased levels of mitochondria-activating factors. We also studied in cultured myocytes and obtained compatible results. ALA-treated mice tended to increase body weight, but reduced blood glucose level. These suggested that ALA treatment not only activated muscle mitochondria but also enhanced muscle mass through an increase in BCAAs contents, as to improve muscle strength, endurance and glucose tolerance in mice. In these ways, muscle mitochondrial activation with ALA is suggested to be useful for the treatment of sarcopenia and glucose intolerance.

Abstract Background Virtual-assisted lung mapping is a novel bronchoscopic lung marking technique that uses virtual images to perform multiple concurrent dye marking of barely palpable pulmonary tumors. Subsequent chest computed tomography is required to confirm the locations marked. We here report a patient in whom computed tomography after virtual-assisted lung mapping unexpectedly revealed additional tiny pulmonary nodules. Case presentation A 64-year-old woman with a history of renal cell carcinoma presented with two pulmonary nodules suspicious of metastases from renal cell carcinoma. Because we anticipated that the nodules would be difficult to palpate intraoperatively, we performed virtual-assisted lung mapping prior to attempting to resect them. Computed tomography after mapping unexpectedly detected two additional nodules. Although the existing markings did not relate to the newly found nodules, we used imaginary auxiliary lines and anatomical landmarks to extend the lung map to incorporate the unexpected nodules. The additional nodules were successfully resected by thoracoscopic wedge resection. Pathologic examination identified all nodules as metastases from renal cell carcinoma, and the surgical margins were negative. Conclusions Imaginary auxiliary lines and anatomical landmarks extended the existing lung map of virtual-assisted lung mapping, enabling resection of unexpected pulmonary nodules found in post-mapping computed tomography images.

Abstract Background Building surgical strategies for complex lung anomalies such as congenital pulmonary venolobar syndrome is difficult because of their rarity and variance. Using three-dimensional computed tomography (3D–CT), we can determine strategies safely. We describe a 27-year-old man with multifocal pulmonary malformations who underwent video-assisted thoracoscopic surgery (VATS) using 3D–CT. Case presentation A 27-year-old man presented with hemoptysis associated with complex pulmonary malformations, including a triple-arched vein connecting the superior and inferior pulmonary veins with partial drainage into the inferior vena cava, a systemic and numerous arterial supply to the right lower lobe from the abdominal aorta, abnormal lobulation, and a tracheal bronchus in the right lung. Preoperative simulation using 3D–CT helped to determine the optimal surgical strategy, allowing for successful and safe completion of semi-emergent middle and lower bilobectomy via minimally invasive VATS. Conclusions Preoperative simulation using 3D–CT helped to determine the optimal surgical strategy, allowing for successful and safe completion of surgery even in a complex case.

In vivo optical imaging is a powerful tool for revealing brain structure and function at both the circuit and cellular levels. Here, we provide a systematic review of findings obtained from in vivo imaging studies of mouse models of neurodevelopmental disorders, including the monogenic disorders fragile X syndrome, Rett syndrome, and Angelman syndrome, which are caused by genetic abnormalities of FMR1, MECP2, and UBE3A, as well as disorders caused by copy number variations (15q11-13 duplication and 22q11.2 deletion) and BTBR mice as an inbred strain model of autism spectrum disorder (ASD). Most studies visualize the structural and functional responsiveness of cerebral cortical neurons to sensory stimuli and the developmental and experience-dependent changes in these responses as a model of brain functions affected by these disorders. The optical imaging techniques include two-photon microscopy of fluorescently labeled dendritic spines or neurons loaded with fluorescent calcium indicators and macroscopic imaging of cortical activity using calcium indicators, voltage-sensitive dyes or intrinsic optical signals. Studies have revealed alterations in the density, stability, and turnover of dendritic spines, aberrant cortical sensory responses, impaired inhibitory function, and concomitant failure of circuit maturation as common causes for neurological deficits. Mechanistic hypotheses derived from in vivo imaging also provide new directions for therapeutic interventions. For instance, it was recently demonstrated that early postnatal administration of a selective serotonin reuptake inhibitor (SSRI) restores impaired cortical inhibitory function and ameliorates the aberrant social behaviors in a mouse model of ASD. We discuss the potential use of SSRIs for treating ASDs in light of these findings.

The recruitment of brite (or beige) adipocytes has been advocated as a means to combat obesity, due to their ability to phenotypically resemble brown adipocytes (BA). Lineage studies indicate that brite adipocytes are formed by differentiation of precursor cells or by direct conversion of existing white adipocytes, depending on the adipose depot examined. We have systematically compared the gene expression profile and a functional output (oxygen consumption) in mouse adipocytes cultured from two contrasting depots, namely interscapular brown adipose tissue, and inguinal white adipose tissue (iWAT), following treatment with a known browning agent, the peroxisome proliferator-activated receptor (PPARγ) activator rosiglitazone. Prototypical BA readily express uncoupling protein (UCP)1, and upstream regulators including the β3-adrenoceptor and transcription factors involved in energy homeostasis. Adipocytes from inguinal WAT display maximal UCP1 expression and mitochondrial uncoupling only when treated with a combination of the PPARγ activator rosiglitazone and a β3-adrenoceptor agonist. In conclusion, brite adipocytes are fully activated only when a browning agent (rosiglitazone) and a thermogenic agent (β3-adrenoceptor agonist) are added in combination. The presence of rosiglitazone throughout the 7-day culture period partially masks the effects of β3-adrenoceptor signaling in inguinal white adipocyte cultures, whereas including rosiglitazone only for the first 3 days promotes robust β3-adrenoceptor expression and provides an improved window for detection of β3-adrenoceptor responses.

Cell-substrate adhesions are a mechanosensitive protein complex that regulates various cellular functions. Molecular mechanisms underlying the physical force-dependent regulation remain elusive partly because endogenous forces are distributed in a spatially heterogeneous manner within cells, thus complicating the interpretation on the effect of forces. Here we use a micropatterning technique to focus spatially distributed intracellular contractile forces onto a particular subcellular area, with which mechanical and pharmacological effects are separately analyzed. Single human osteosarcoma U2OS cells were plated within square micropatterns, and phosphorylation of an adhesion-associated adaptor protein paxillin was analyzed. Paxillin, visualized with immunostaining, was highly accumulated in the proximity of the corners of the square micropatterns where cellular forces are concentrated, but the huge paxillin-labeled adhesions were less phosphorylated compared to those present elsewhere as a small patch. Pharmacological inhibition of the endogenous forces resulted in disassembly of the huge dephoshorylated paxillin clusters; in contrast, the small, highly phosphorylated paxillin patches persisted. Similar negative regulation of paxillin phosphorylation is also induced upon loading of exogenous forces. Unexpectedly, on the other hand, immunoblot of cell lysates showed a tendency of a reduction in paxillin phosphorylation upon the same pharmacological inhibition of the endogenous forces. Thus, the response of paxillin phosphorylation to mechanical forces was the opposite between the immunostaining and immunoblot data; i.e., the phosphorylation is reduced and enhanced at subcellular level and whole-cell level, respectively, in response to loading of mechanical forces. To reconcile the contradictory results, we submit a simple model that is consistent with not only the present but also previous reports on the regulation of paxillin. The model implies that similar opposite response can generally emerge if the protein activation is negatively regulated at a local place while the activation trigger alters the assembly of the protein to the local place.

Fluid shear stress (SS) is well known to cause morphological changes in vascular endothelial cells (ECs) accompanied by alteration in actin cytoskeletal structure and distribution of focal adhesions. Recent studies have shown that spatial SS gradient also has effects on EC morphology, but the detailed mechanisms of EC responses to SSG remain unclear. In the present study, we sought morphological responses of ECs under SS and uniform SSG condition using a newly developed flow chamber. Confluent ECs were exposed to SS with SSG for 24 hours. Focal adhesions of the EC under SS without SSG were localized in the cell periphery. In contrast, focal adhesions were expressed not only in the periphery but also in interior portion of cells after exposure to SS with SSG. Unlike ECs exposure to SS developed thick actin filaments aligned to the direction of flow no development of thick actin filaments but thin and short filaments were observed in ECs after 24-hour exposure to SS with SSG. Since the distribution of focal adhesion is of critical importance for development of actin filaments and cell morphological changes, these results suggest that SSG suppresses redistribution of focal adhesions, resulting in the inhibition of EC morphological changes and development of thick actin filaments in response to flow.

Actin stress fibers (SFs) play a key role in regulation of cell adhesion, but the biochemical and biophysical properties intrinsic to SFs remain unclear. Here we extracted SFs from rat embryonic smooth muscle cells by deroofing, and evaluated the effects of varying ionic strength and temperature on their intactness. Wash buffers with ionic strength ranging from 90 to 490 mM were prepared, and the extracted SFs were incubated in a buffer with a particular ionic strength for 10 min or 24 h. Light and electron microscopy revealed that the extracted SFs comprised tightly packed straight bundles at low ionic strengths that became looser and exhibited a ragged pattern at high ionic strengths. The expression of α-actinin associated with the extracted SFs decreased with the increase in ionic strength. Unexpectedly, non-muscle myosin II and smooth muscle myosin in the extracted SFs displayed comparable expression levels over the different ionic strengths. ATP-induced contractility was better preserved at low ionic strengths, including the physiological ionic strength of 170 mM. The rate of ATP-induced enzymatic activity increased with increase in temperature, but the difference was not statistically significant. These results demonstrate that low ionic strength produces extracted SFs that are more intact with regard to structure and function.

Cellular traction forces were measured by using a microfabricated substrate, particularly exploring how cytoskeletal structures such as actin filaments and microtubules contribute to traction forces. Smooth muscle cells isolated from bovine aortas were cultured and transfected with fluorescence proteins to visualize cell microstructures and then plated on a micropatterned elastomer substrate with arrays of micropillars. Cell spreading on the substrates produced deflection of micropillars which was used for estimation of cellular traction forces, and was closely associated with organization of stress fibers of actin filaments. Traction forces varied considerably among cells, showing the order of several 10 nN. After disruption of microtubules with nocodazole, traction forces significantly increased and there was no detectable change in formation of stress fibers. To inhibit the ROCK pathway, a signaling pathway of myosin light chain phosphorylation, possibly being induced by disruption of microtubules, significantly depressed the increase in traction forces after the disruption of microtubules. This result indicates that microtubules disassembly may regulate the actomyosin-based contractile system mainly through the ROCK pathway. The present study suggests that formation of stress fibers are mainly involved in cellular traction forces and a contribution of microtubules should include not only a force balance but also rather a modulator of the actomyosin contractile system in actin stress fibers.

Mechanical properties of human aortic aneurysm tissues were measured with a biaxial tensile tester. Fifteen-mm-square specimens were obtained from thoracic aortic aneurysms of various origins and from undilated aortas adjacent to the aneurysms during aneurysmectomy, and were stored frozen until the measurement. Each specimen was stretched biaxially in physiological saline at room temperature at the rate of ∼0.2 mm/sec. Although the ordered displacement was set equal for both directions, real strain applied to the specimens was not equibiaxial. The stress-strain curves under equibiaxial stretch were obtained by fitting measured curves with a strain energy function considering material anisotropy. Effects of freezing and ambient temperature on the mechanical properties were evaluated with porcine thoracic aortas. The mechanical properties of the frozen-stored specimens at 23°C were almost similar to those of the fresh specimens at 37 °C. Elastic modulus at zero load averaged for both directions Hmi = (Hxi+Hyi)/2 was higher (P < 0.01) in the aneurysm tissues (1450 ± 250 kPa, mean ± SEM, n = 26) than in the undilated tissues (650 ± 140 kPa, n = 10). Anisotropy index K = |Hxi-Hyi|/Hmi was not significantly different between the aneurysm (20 ± 3%) and the undilated tissues (12 ± 3%) for all specimens. For the specimens whose elastic modulus Hmi was smaller than 1 MPa, however, the index K was significantly higher (P < 0.05) in the aneurysm specimens (23.1 ± 5.3%, n = 14) than the undilated tissues (9.5 ± 2.5%, n = 8). These results indicate aneurysm tissues are not only stiffer but also more anisotropic than the nonaneurysmal tissues.

Control of the gliding directions of kinesin-driven microtubules (MTs) in vitro has good feasibility for the development of nano-scale transport systems. A requirement for the control of transporters in these systems includes detecting the positions of gliding MTs; however, no studies have reported on the monitoring of the positions of gliding MTs. Here, we suggest an algorithm to detect tip coordinates of gliding MTs by binarization, skeletonization, and filtration of fluorescent images of MTs. The algorithm was first applied to artificially drawn segments with given lengths (10-80 pixels), widths (1-10 pixels), and curvature radii (20-120 pixels) to verify the effect of the sizes of MTs on accuracy of tip coordinates extracted by the algorithm, and error was estimated by referring to the true coordinates. The estimated errors were as small as 2 pixels in the width and were not affected by the length and the curvature radius, indicating that our algorithm is useful to extract the tips of MTs. The algorithm was subsequently applied to images of gliding MTs. Since distances from the trajectories of the MTs to the centers of gravity of the MTs (3.7 ± 2.1 pixels) were significantly larger than those to the tips (1.9 ± 0.5 pixels), the use of the tips as representative points of gliding MTs was verified. A detection method using tips of MTs, as suggested in this study, may be a useful technique for monitoring each MT in nanoscale transport systems.