Publications by authors named "Masahiko Kobayashi"

139 Publications

Arthroscopic evaluation after osteochondral autogenous transfer with osteotomy of medial malleolus for osteochondral lesion of the talar dome.

Foot Ankle Surg 2021 Feb 4. Epub 2021 Feb 4.

Department of Orthopedic Surgery, Kyoto Katsura Hospital, Japan.

Background: The purpose of this study was to investigate the second-look arthroscopic evaluation after osteochondral autogenous transfer (OAT) for osteochondral lesion of the talar dome (OLT) with the criteria of the International Cartilage Repair Society (ICRS).

Methods: Ten patients (twelve ankles) with OLT underwent OAT with osteotomy of the medial malleolus. Clinical outcomes were evaluated using the American Orthopedic Foot & Ankle Society (AOFAS) ankle-hindfoot scale. The condition of the transplanted cartilage was evaluated at the time of second-look arthroscopy using the ICRS Cartilage Repair Assessment.

Results: The AOFAS ankle-hindfoot scale was significantly improved from 65.1 ± 1.9 points before surgery to 98.1 ± 2.8 points at the time of second-look arthroscopy (p < 0.01). The ICRS Cartilage Repair Assessment was 11.4 points on average (9-12 points).

Conclusions: The OAT for OLT is considered to be a useful treatment even if invasion by medial malleolus osteotomy is added.

Level Of Evidence: Level IV, Case series.
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http://dx.doi.org/10.1016/j.fas.2021.01.005DOI Listing
February 2021

Essential role of autophagy in protecting neonatal haematopoietic stem cells from oxidative stress in a p62-independent manner.

Sci Rep 2021 Jan 18;11(1):1666. Epub 2021 Jan 18.

Division of Molecular Genetics, Cancer and Stem Cell Research Program, Cancer Research Institute, Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, 920-1192, Japan.

Autophagy is a cellular degradation system contributing to homeostasis of tissue stem cells including haematopoietic stem cells (HSCs). It plays pleiotropic roles in HSC characteristics throughout life, but its stage-specific roles in HSC self-renewal are unclear. To investigate the effects of Atg5 deletion on stage-specific HSC functions, we compared the repopulating capacity of HSCs in Atg5;Vavi-cre mice from postnatal day (P) 0-7 weeks of age. Interestingly, Atg5 deficiency led to no remarkable abnormality in the HSC self-renewal capacity at P0, but significant defects at P7, followed by severe defects. Induction of Atg5 deletion at P5 by tamoxifen administration to Atg5;Rosa26-Cre-ER mice resulted in normal haematopoiesis, including the HSC population, until around 1 year, suggesting that Atg5 in the early neonatal period was critical for haematopoiesis in adults. Mitochondrial oxidative stress was increased by Atg5 loss in neonatal HSC/progenitor cells. Although p62 had accumulated in immature bone marrow cells of Atg5;Vavi-cre mice, p62 deletion did not restore defective HSC functions, indicating that Atg5-dependent haematopoietic regulation in the developmental period was independent of p62. This study proposes a critical role of autophagy in HSC protection against harsh environments in the early neonatal stage, which is essential for healthy long-term haematopoiesis.
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http://dx.doi.org/10.1038/s41598-021-81076-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7814027PMC
January 2021

Long-Term Survivorship and Clinical Outcomes of Osteochondral Autologous Transplantation for Steroid-Induced Osteonecrosis of the Knee.

Cartilage 2020 Sep 10:1947603520954489. Epub 2020 Sep 10.

Department of Orthopaedic Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan.

Objective: Treatments for steroid-induced osteonecrosis of the knee remains challenging, and there has not been sufficient evidence to support joint preservation surgery. This study evaluated long-term outcomes of osteochondral autologous transplantation (OAT) for steroid-induced osteonecrosis of the knee.

Design: This retrospective case series included patients who underwent OAT for steroid-induced osteonecrosis of the knee from 1998 to 2008. The survivorship and need for secondary surgery were evaluated, and the clinical outcome was evaluated with the International Knee Documentation Committee (IKDC) subjective score. Preoperative and final Kellgren-Lawrence (KL) grade of the femorotibial and patellofemoral joints were individually evaluated.

Results: Fourteen knees of 10 patients whose mean age was 32.5 (95%CI 26.4-38.6) years were included and followed for 14.0 (12.4-15.7) years. The mean lesion size of 6.9 (5.3-8.5) cm was repaired using 4 median (minimum 2, maximum 5) osteochondral plugs. No revision surgeries were performed for transplanted osteochondral plugs. The IKDC subjective score improved from 32.9 (24.5-41.3) to 74.2 (61.9-88.5) ( < 0.001). Knee flexion was improved at the final follow-up, and Seiza sitting was finally possible in 9 knees in 7 patients. Although the osteoarthritic change did not progress in femorotibial joint, patellofemoral joint showed early osteoarthritic changes at the final follow-up (mean KL grade: 0.8 [0.5-1.1]).

Conclusions: Prosthetic joint replacement was successfully avoided for at least the first decade by OAT in young patients with steroid-induced osteonecrosis of the knee. The progression of KL grade of the patellofemoral joint is of concern.
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http://dx.doi.org/10.1177/1947603520954489DOI Listing
September 2020

Etiologies of pericarditis in hospital and forensic autopsies.

Cardiovasc Pathol 2020 Nov - Dec;49:107262. Epub 2020 Jul 10.

Department of Pathology, John A. Burns School of Medicine, University of Hawaii, 651 Ilalo Street, Honolulu, HI 96813, USA; The Queen's Medical Center, 1301 Punchbowl Street, Honolulu, HI 96813, USA.

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http://dx.doi.org/10.1016/j.carpath.2020.107262DOI Listing
November 2020

Does TMJ Function and Imaging Tools Help Differentiate Between Condylar Resorption and Mandibular Hypoplasia?

J Oral Maxillofac Surg 2020 Aug 11;78(8):1397-1402. Epub 2020 Apr 11.

Professor and Chair, Division of Oral and Maxillofacial Surgery, Department of Diagnostic and Therapeutic Sciences, Meikai University School of Dentistry, Saitama, Japan.

Purpose: Differentiating between bilateral condylar resorption (CR) and mandibular hypoplasia (MH) can be challenging owing to the difficulty in chronological observation and establishing standardized measurements. The purpose of the present study was to assess whether temporomandibular joint (TMJ) function can distinguish between CR and MH and clarify the essential diagnostic imaging tools for CR.

Materials And Methods: We performed a cross-sectional study of patients with mandibular retrognathia. The primary predictor variables were a clinical dysfunction score for the TMJ, mandibular plane angle (MPA), SNA angle, SNB angle, and cortical erosion score in the condylar heads. The demographic variables were age, anterior disc displacement, and previous orthodontic treatment. The anatomic variable was the condylar height (CH). The primary outcome variable was the disease status (CR or MH). The patients were divided into the CR group and MH group. The patients with CR were selected on the basis of a CH value of less than 22 mm. TMJ function was assessed using the Helkimo clinical dysfunction index. The CH on panoramic radiographs was measured using the Kjellberg method. The MPA, SNA angle, and SNB angle were analyzed using cephalometric analysis. Cortical erosion in the condylar head was assessed using computed tomography and magnetic resonance imaging.

Results: A total of 23 female participants were enrolled in the present study. The average clinical dysfunction score for the TMJ was 4.4 in the CR group and 0.4 in the MH group (P < .05). The average MPA was 41.2° in the CR group and 35.5° in the MH group (P < .05).

Conclusions: The present investigation has shown that assessing TMJ function and analyzing MPA using a cephalometric radiograph can differentiate CR from MH.
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http://dx.doi.org/10.1016/j.joms.2020.04.002DOI Listing
August 2020

Identification of 2-Fluoropalmitic Acid as a Potential Therapeutic Agent Against Glioblastoma.

Curr Pharm Des 2020 ;26(36):4675-4684

Department of Neurosurgery, Graduate School of Medical Science, Kanazawa University, Kanazawa, Japan.

Background: Glioblastomas (GBMs) are aggressive malignant brain tumors. Although chemotherapy with temozolomide (TMZ) can extend patient survival, most patients eventually demonstrate resistance. Therefore, novel therapeutic agents that overcome TMZ chemoresistance are required to improve patient outcomes.

Purpose: Drug screening is an efficient method to find new therapeutic agents from existing drugs. In this study, we explored a novel anti-glioma agent by drug screening and analyzed its function with respect to GBM treatment for future clinical applications.

Methods: Drug libraries containing 1,301 diverse chemical compounds were screened against two glioma stem cell (GSC) lines for drug candidate selection. The effect of selected agents on GSCs and glioma was estimated through viability, proliferation, sphere formation, and invasion assays. Combination therapy was performed to assess its ability to enhance TMZ cytotoxicity against GBM. To clarify the mechanism of action, we performed methylation-specific polymerase chain reaction, gelatin zymography, and western blot analysis.

Results: The acyl-CoA synthetase inhibitor 2-fluoropalmitic acid (2-FPA) was selected as a candidate anti-glioma agent. 2-FPA suppressed the viability and stem-like phenotype of GSCs. It also inhibited proliferation and invasion of glioma cell lines. Combination therapy of 2-FPA with TMZ synergistically enhanced the efficacy of TMZ. 2-FPA suppressed the expression of phosphor-ERK, CD133, and SOX-2; reduced MMP-2 activity; and increased methylation of the MGMT promoter.

Conclusion: 2-FPA was identified as a potential therapeutic agent against GBM. To extend these findings, physiological studies are required to examine the efficacy of 2-FPA against GBM in vivo.
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http://dx.doi.org/10.2174/1381612826666200429092742DOI Listing
March 2021

Identification of GSK3β inhibitor kenpaullone as a temozolomide enhancer against glioblastoma.

Sci Rep 2019 07 11;9(1):10049. Epub 2019 Jul 11.

Department of Neurosurgery, Faculty of Medicine, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kanazawa, Japan.

Cancer stem cells are associated with chemoresistance and rapid recurrence of malignant tumors, including glioblastoma (GBM). Although temozolomide (TMZ) is the most effective drug treatment for GBM, GBM cells acquire resistance and become refractory to TMZ during treatment. Therefore, glioma stem cell (GSC)-targeted therapy and TMZ-enhancing therapy may be effective approaches to improve GBM prognosis. Many drugs that suppress the signaling pathways that maintain GSC or enhance the effects of TMZ have been reported. However, there are no established therapies beyond TMZ treatment currently in use. In this study, we screened drug libraries composed of 1,301 existing drugs using cell viability assays to evaluate effects on GSCs, which led to selection of kenpaullone, a kinase inhibitor, as a TMZ enhancer targeting GSCs. Kenpaullone efficiently suppressed activity of glycogen synthase kinase (GSK) 3β. Combination therapy with kenpaullone and TMZ suppressed stem cell phenotype and viability of both GSCs and glioma cell lines. Combination therapy in mouse models significantly prolonged survival time compared with TMZ monotherapy. Taken together, kenpaullone is a promising drug for treatment of GBM by targeting GSCs and overcoming chemoresistance to TMZ.
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http://dx.doi.org/10.1038/s41598-019-46454-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6624278PMC
July 2019

Adhesion and Proliferation of Osteoblastic Cells on Hydroxyapatite-dispersed Ti-based Composite Plate.

In Vivo 2019 Jul-Aug;33(4):1067-1079

Division of First Oral and Maxillofacial Surgery, Meikai University School of Dentistry, Sakado, Japan.

Background/aim: Biocompatibility of a novel and more stable hydroxyapatite (HA)-dispersed titanium (Ti)-based composite was investigated, using the mouse osteoblast precursor cell line MC3T3-E1.

Materials And Methods: Surface of powders and plates was observed by scanning electron microscopy. Distribution of calcium and phosphorus on the surface of the composite was evaluated by an electron beam microanalyzer. Crystal structure was analyzed by X-ray diffractometer. Cell viability was determined by WST-1 assay.

Results: HA was stable against the compressive force, shearing stress and sintering heat at 800°C, but it slightly decomposed at 1100°C. With the increase of HA in the composites, the adhesion/proliferation of MC3T3-E1 cells was reduced. The growth inhibition by HA does not seem to be due to materials released from the plate, but rather to the contact to the surface of the plate. Sintering of the HA plate at 1100°C increased the number of attached viable cells. On the other hand, culturing on the synthesized calcium phosphate (apatite containing carbonic acid) increased the number of attached cells to a greater extent.

Conclusion: HA inhibits the growth of osteoblastic cells, but sintering at 1100°C changes the surface properties of the composite so as to stimulate cell growth.
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http://dx.doi.org/10.21873/invivo.11575DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6689340PMC
December 2019

[Survey of the Implementation Status of Personal Counseling among Cancer Patients in Red Cross Hospitals].

Gan To Kagaku Ryoho 2019 May;46(5):907-911

Dept. of Pharmacy, Matsuyama Red Cross Hospital.

Red Cross Pharmaceutical Association conducted a questionnaire-based survey to evaluate the status of implementation of pharmaceutical intervention as well as personal counseling for outpatients with cancer undergoing chemotherapy. Based on the survey results from 93 hospitals across the country, it was found that pharmacists performed an intervention on outpatients receiving chemotherapy in 68 hospitals(73.1%)and conducted personal counseling for outpatients with cancer in 48 hospitals(51.6%). Out of the 68 hospitals, 20 did not conduct personal counseling for outpatients with cancer. This was attributable to the fact that 14 hospitals did not have a qualified pharmacist, 3 did not have sufficient manpower, and 3 did not have the required system. The results of a logistic regression analysis showed that the number of pharmacists significantly affected implementation of pharmaceutical intervention as well as personal counseling for outpatients with cancer undergoing chemotherapy(p=0.042, p=0.023, respectively). The pharmacists can receive a fee for medical services only after conducting personal counseling for outpatients with cancer undergoing chemotherapy. However, in hospitals with a small number of pharmacists, they could not claim their fees owing to lack of manpower. This survey found that lack of manpower is currently the most important issue.
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May 2019

Clinical and Radiographic Outcomes After Arthroscopic Lamina-Specific Double-Row Repair of Large Delaminated Rotator Cuff Tears in Active Patients.

Orthop J Sports Med 2019 Apr 22;7(4):2325967119838249. Epub 2019 Apr 22.

Department of Orthopaedic Surgery, Kyoto Shimogamo Hospital, Kyoto, Japan.

Background: The presence of delamination and a larger rotator cuff tear (RCT) size have been associated with poorer outcomes in rotator cuff repair. Therefore, we developed a new surgical procedure, arthroscopic lamina-specific double-row fixation (ALSDR), for the repair of large delaminated RCTs.

Purpose: To investigate the clinical outcomes, magnetic resonance imaging findings, and satisfaction with several variables after ALSDR for large delaminated RCTs.

Study Design: Case series; Level of evidence, 4.

Methods: A total of 30 active patients (mean age, 59.1 years) undergoing ALSDR were assessed by a numeric rating scale (NRS; 0-10) for pain, surgery, work, and exercise as well as American Shoulder and Elbow Surgeons (ASES), Constant, and Simple Shoulder Test (SST) scores at a mean of 65.9 months postoperatively. Rotator cuff integrity was determined by magnetic resonance imaging. The Spearman correlation coefficient (ρ) was used to determine the correlation between clinical and NRS scores.

Results: Five patients (16.7%) had a retear. Each of the postoperative functional and NRS scores except the NRS work score was significantly better in the healed shoulders than in the shoulders with a retear ( < .001). The NRS pain score showed a significant negative correlation with ASES, Constant, and SST scores (ρ = -0.775, -0.668, and -0.742, respectively; < .001 for all). The NRS surgery score had a positive correlation with Constant and SST scores (ρ = 0.393 [ = .032] and ρ = 0.456 [ = .011], respectively). The NRS work score had a positive correlation with ASES, Constant, and SST scores (ρ = 0.382 [ = .037], ρ = 0.386 [ = .035], and ρ = 0.414 [ = .023], respectively). The NRS exercise score had a positive correlation with ASES, Constant, and SST scores (ρ = 0.567 [ = .001], ρ = 0.511 [ = .004], and ρ = 0.639 [ < .001], respectively).

Conclusion: Our results showed that there was a significant correlation between clinical and NRS scores. The results indicate that ALSDR can provide a high degree of functionality and can be a useful alternative treatment for active patients with large delaminated RCTs.
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http://dx.doi.org/10.1177/2325967119838249DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6477774PMC
April 2019

Radial incision and cutting for releasing severe stricture enables successful delivery of an over-the-scope clip.

Endoscopy 2019 07 12;51(7):E179-E180. Epub 2019 Apr 12.

Department of Gastroenterology and Neurology, Faculty of Medicine, Kagawa University, Kagawa, Japan.

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http://dx.doi.org/10.1055/a-0871-1749DOI Listing
July 2019

Bone healing potential of fascia lata autografts to the humeral head footprint in rotator cuff reconstruction based on magnetic resonance imaging and histologic evaluations.

J Shoulder Elbow Surg 2019 Jul 1;28(7):1363-1370. Epub 2019 Mar 1.

Department of Orthopaedic Surgery, Kyoto Shimogamo Hospital, Kyoto, Japan.

Background: The purpose of the study was to evaluate the bone healing potential of fascia lata autograft (FLA) by magnetic resonance imaging (MRI) and histologic analysis.

Methods: The study included 69 patients assessed by MRI after an FLA patch procedure. Three of the 69 patients underwent a revision procedure after the primary FLA procedure; 1 underwent a second-look arthroscopy and 2 underwent reverse shoulder arthroplasties (RSAs). In the 2 RSA patients, we histologically evaluated greater tuberosities with the repaired graft. Moreover, as a control, we harvested the greater tuberosity with the cuff tendon at the time of RSA for failed open reduction-internal fixation of 4-part proximal humeral fracture. Based on MRI, retear cases were divided into type 1 (the graft did not remain on the greater tuberosity) and type 2 (the graft remained on the greater tuberosity). Histologic sections were evaluated to examine fascia-bone or rotator cuff-bone interfaces.

Results: There were 35 intact repairs: 7 type 1 and 27 type 2 shoulders (type 1 vs. type 2, P < .001). Second-look arthroscopic findings confirmed that the graft was securely attached to the greater tuberosity. Histologic analysis of greater tuberosities in RSA patients showed solid continuity of the graft to the bone, with cells with nuclei in the collagen matrix oriented in parallel. The FLA to bone junction consisted of the FLA, fibrocartilage, and bone, which is similar to the normal cuff tendon to bone junction.

Conclusions: These results indicate that a fresh cellular FLA has good to excellent bone healing potential.
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http://dx.doi.org/10.1016/j.jse.2018.11.067DOI Listing
July 2019

Hyperemia Analysis Software for Assessment of Conjunctival Hyperemia Severity.

Curr Eye Res 2019 04 12;44(4):376-380. Epub 2018 Dec 12.

b Department of Ophthalmology , Kochi Medical School , Nankoku-city , Kochi , Japan.

Purpose: We developed a hyperemia analysis software, which can quantitatively assess the degree of conjunctival hyperemia, and evaluated the reproducibility and reliability of its percent coverage. In addition, we compared the clinical grading and the percent coverage to examine the applicability of the software analysis.

Methods: We took images of the temporal conjunctiva with slit lamp microscopes. We used our hyperemia analysis software to detect blood vessels in the region of interest using its image processing capabilities and obtained a ratio of the area occupied by blood vessels to the rest of the area. In addition, we used the clinical evaluation criteria of the Japanese guidelines for allergic conjunctival diseases 2017 to clinically grade the hyperemia in each eye. Nine evaluators examined the images and graded the severity of hyperemia into four stages. We looked for a statistical correlation between the results of the hyperemia analysis software and the clinical grading.

Results: The percent coverage of the blood vessels in the region of interest calculated by the hyperemia analysis software correlated with the arithmetic average of our clinical grading (r = 0.953; 95% CI, 0.8470340-0.9862136).

Conclusion: The percent coverage from our hyperemia analysis software reflects the clinical grading score, suggesting that our software can be used to obtain a detailed analysis of conjunctival hyperemia.
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http://dx.doi.org/10.1080/02713683.2018.1554153DOI Listing
April 2019

Anatomic Coracoclavicular Ligament Reconstruction for the Treatment of Acute Acromioclavicular Joint Dislocation: Minimum 10-Year Follow-up.

JB JS Open Access 2017 Sep 10;2(3):e0007. Epub 2017 Aug 10.

Kyoto Shimogamo Hospital, Kyoto, Japan.

Background: The long-term clinical and radiographic outcomes following coracoclavicular (CC) ligament reconstruction for the operative treatment of acute acromioclavicular (AC) joint dislocation remain uncertain. The purpose of the present study was to determine the long-term clinical and radiographic outcomes of CC ligament reconstruction and to identify risk factors for unfavorable outcomes.

Methods: We reviewed 20 cases of AC joint dislocation in 19 patients (18 male and 1 female; mean age, 32.3 years) that were treated with single-bundle reconstruction. The mean duration of follow-up was 12.7 years. We measured the CC vertical distance (CCD) on the anteroposterior view and compared the affected and unaffected sides (CCD ratio). We divided the patients into those with a CCD ratio of <25% (Group 1) and those with a CCD ratio of ≥25% (Group 2). We radiographically investigated the clavicular tunnel anteroposterior (CTAP) angle, clavicular tunnel ratio, and coracoid tunnel orientation on the basis of the entry and exit points at the base of the coracoid. For the coracoid tunnel orientation, we compared center-center orientation and noncenter-center orientation.

Results: Group 1 comprised 17 cases (85%), and Group 2 comprised 3 cases (15%). At the time of the latest follow-up, Group 1 had a significantly higher mean Constant score than Group 2 (98.2 compared with 90.7; p = 0.038). Of the 3 radiographic parameters, only the CTAP angle was significantly different between the 2 groups (p < 0.0001). Two (67%) of the 3 cases in Group 2 were associated with posterior AC joint displacement.

Conclusions: CC ligament reconstruction for the treatment of acute AC joint dislocation resulted in successful long-term clinical and radiographic outcomes. It is important to decrease the CTAP angle and to ensure proper anatomic placement of the clavicular and coracoid tunnels at the time of surgery.

Level Of Evidence: Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.
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http://dx.doi.org/10.2106/JBJS.OA.16.00007DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6133097PMC
September 2017

Vocal practice regulates singing activity-dependent genes underlying age-independent vocal learning in songbirds.

PLoS Biol 2018 09 12;16(9):e2006537. Epub 2018 Sep 12.

Graduate School of Life Science, Hokkaido University, Sapporo, Hokkaido, Japan.

The development of highly complex vocal skill, like human language and bird songs, is underlain by learning. Vocal learning, even when occurring in adulthood, is thought to largely depend on a sensitive/critical period during postnatal development, and learned vocal patterns emerge gradually as the long-term consequence of vocal practice during this critical period. In this scenario, it is presumed that the effect of vocal practice is thus mainly limited by the intrinsic timing of age-dependent maturation factors that close the critical period and reduce neural plasticity. However, an alternative, as-yet untested hypothesis is that vocal practice itself, independently of age, regulates vocal learning plasticity. Here, we explicitly discriminate between the influences of age and vocal practice using a songbird model system. We prevented zebra finches from singing during the critical period of sensorimotor learning by reversible postural manipulation. This enabled to us to separate lifelong vocal experience from the effects of age. The singing-prevented birds produced juvenile-like immature song and retained sufficient ability to acquire a tutored song even at adulthood when allowed to sing freely. Genome-wide gene expression network analysis revealed that this adult vocal plasticity was accompanied by an intense induction of singing activity-dependent genes, similar to that observed in juvenile birds, rather than of age-dependent genes. The transcriptional changes of activity-dependent genes occurred in the vocal motor robust nucleus of the arcopallium (RA) projection neurons that play a critical role in the production of song phonology. These gene expression changes were accompanied by neuroanatomical changes: dendritic spine pruning in RA projection neurons. These results show that self-motivated practice itself changes the expression dynamics of activity-dependent genes associated with vocal learning plasticity and that this process is not tightly linked to age-dependent maturational factors.
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http://dx.doi.org/10.1371/journal.pbio.2006537DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6152990PMC
September 2018

Confocal Raman microscopy reveals changes in chemical composition of wood surfaces exposed to artificial weathering.

J Photochem Photobiol B 2018 Oct 16;187:136-140. Epub 2018 Aug 16.

College of Bioresource Sciences, Nihon University, Kameino 1866, Fujisawa, Kanagawa 252-0880, Japan.

Weathering leads to rapid depolymerization and modification of wood chemical components. The present study aims to assess the structural and distributional changes in the main wood polymers, such as lignin and polysaccharides, located in the surface layers during weathering exposure. A confocal Raman microscopic technique, which is useful for evaluating the structure of molecules with a high spatial resolution, was utilized to examine the effects of weathering on the chemical composition of wood surfaces at the cellular level. Raman spectra showed that lignin degradation during weathering proceeds with the formation of o- and p-quinones, carbonyl groups, and certain types of CC double bonds such as stilbene derivatives. Comparing the weathering conditions between light only and light plus water exposure, it was found that weathering in the presence of water significantly enhances the degradation of lignin. The Raman images exhibited that the degree of lignin degradation by light only exposure is according to the depth. However, in the case of light plus water exposure, lignin degradation in the outermost cell walls proceeded from both the exposed surface side and the lumen side of the cell walls. This study is expected to potentially promote development of more effective and efficient methods to protect wood surfaces against weathering.
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http://dx.doi.org/10.1016/j.jphotobiol.2018.08.016DOI Listing
October 2018

Composite neuroendocrine carcinoma and squamous cell carcinoma with regional lymph node metastasis: a case report.

J Med Case Rep 2018 Aug 24;12(1):227. Epub 2018 Aug 24.

Department of Gastroenterology and Neurology, Faculty of Medicine, Kagawa University, Miki, Kagawa, Japan.

Background: Neuroendocrine cell carcinoma is a rare variant of esophageal carcinoma. The characteristic clinical features and diagnosis of superficial neuroendocrine cell carcinoma remain to be established. We report a rare case of superficial coexistence of neuroendocrine cell carcinoma with squamous cell carcinoma treated by endoscopic submucosal dissection, and regional lymph node metastasis was detected after additional surgical treatment.

Case Presentation: A 77-year-old Japanese man with esophageal squamous cell carcinoma received endoscopic submucosal dissection in en-bloc resection. Histopathological findings showed that lymphovascular invasion by the neuroendocrine cell carcinoma component occurred in the deep part of the muscularis mucosa. Regional lymph node metastasis was identified after additional surgical treatment. After surgical treatment, our patient received chemotherapy consisting of etoposide and carboplatin for 3 months. He is alive and shows no sign of disease recurrence 12 months after surgery.

Conclusions: This case report highlights the fact that even if neuroendocrine cell carcinoma is small and limited to superficial, the tumor has the potential for metastasis if lymphovascular invasion by the neuroendocrine cell carcinoma component occurs. In addition, this case indicates the necessity of close follow-up of small neuroendocrine cell carcinoma after treatment.
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http://dx.doi.org/10.1186/s13256-018-1775-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6108124PMC
August 2018

Autophagy inhibition synergizes with calcium mobilization to achieve efficient therapy of malignant gliomas.

Cancer Sci 2018 Aug 26;109(8):2497-2508. Epub 2018 Jul 26.

Cancer and Stem Cell Research Program, Division of Molecular Genetics, Cancer Research Institute, Kanazawa University, Kanazawa, Ishikawa, Japan.

Autophagy plays a critical role in tumorigenesis, but how autophagy contributes to cancer cells' responses to chemotherapeutics remains controversial. To investigate the roles of autophagy in malignant gliomas, we used CRISPR/CAS9 to knock out the ATG5 gene, which is essential for autophagosome formation, in tumor cells derived from patients with glioblastoma. While ATG5 disruption inhibited autophagy, it did not change the phenotypes of glioma cells and did not alter their sensitivity to temozolomide, an agent used for glioblastoma patient therapy. Screening of an anticancer drug library identified compounds that showed greater efficacy to ATG5-knockout glioma cells compared to control. While several selected compounds, including nigericin and salinomycin, remarkably induced autophagy, potent autophagy inducers by mTOR inhibition did not exhibit the ATG5-dependent cytoprotective effects. Nigericin in combination with ATG5 deficiency synergistically suppressed spheroid formation by glioma cells in a manner mitigated by Ca chelation or CaMKK inhibition, indicating that, in combination with autophagy inhibition, calcium-mobilizing compounds contribute to efficient anticancer therapeutics. ATG5-knockout cells treated with nigericin showed increased mitochondria-derived reactive oxygen species and apoptosis compared to controls, indicating that autophagy protects glioma cells from mitochondrial reactive oxygen species-mediated damage. Finally, using a patient-derived xenograft model, we demonstrated that chloroquine, a pharmacological autophagy inhibitor, dramatically enhanced the efficacy of compounds selected in this study. Our findings propose a novel therapeutic strategy in which calcium-mobilizing compounds are combined with autophagy inhibitors to treat patients with glioblastoma.
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http://dx.doi.org/10.1111/cas.13695DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6113445PMC
August 2018

JLP-JNK signaling protects cancer cells from reactive oxygen species-induced cell death.

Biochem Biophys Res Commun 2018 06;501(3):724-730

Division of Molecular Cell Signaling, Cancer Research Institute, Kanazawa University, Kanazawa, Japan. Electronic address:

Oxidative stress, which can be caused by an overproduction of reactive oxygen species (ROS), often leads to cell death. In recent years, c-Jun NH-terminal kinase (JNK)-associated leucine zipper protein (JLP, also known as SPAG9 or JIP4), a scaffold protein for JNK mitogen-activated protein kinase (MAPK) signaling pathways, was found to serve as a novel biomarker for cancer. However, although JNK MAPK pathways are reported to be activated in response to various stimuli, including oxidative stress, whether JLP is involved in ROS signaling remains unknown. In this study, we examined the role of JLP in hydrogen peroxide (HO)-induced cancer cell death, and found that JLP knockdown (KD) cells exhibit a substantially enhanced cell death response, along with increased intracellular ROS levels. This is the first demonstration of a protective role for JLP in response to cell-death stimulation. We also found that the HO-induced JNK activation was attenuated in JLP KD cancer cells. The decreases in cell viability and JNK activation in the JLP KD cells were almost completely reversed by expressing wild-type JLP, but not a mutant JLP lacking the JNK-binding domain. These data collectively suggest that the JLP-JNK signaling pathway counteracts ROS-induced cancer cell death.
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http://dx.doi.org/10.1016/j.bbrc.2018.05.055DOI Listing
June 2018

Spred1 Safeguards Hematopoietic Homeostasis against Diet-Induced Systemic Stress.

Cell Stem Cell 2018 05 26;22(5):713-725.e8. Epub 2018 Apr 26.

Division of Molecular Genetics, Cancer and Stem Cell Research Program, Cancer Research Institute, Kanazawa University, Kanazawa, Ishikawa 920-1192, Japan; WPI Nano Life Science Institute (WPI-Nano LSI), Kanazawa University, Kanazawa, Ishikawa 920-1192, Japan. Electronic address:

Stem cell self-renewal is critical for tissue homeostasis, and its dysregulation can lead to organ failure or tumorigenesis. While obesity can induce varied abnormalities in bone marrow components, it is unclear how diet might affect hematopoietic stem cell (HSC) self-renewal. Here, we show that Spred1, a negative regulator of RAS-MAPK signaling, safeguards HSC homeostasis in animals fed a high-fat diet (HFD). Under steady-state conditions, Spred1 negatively regulates HSC self-renewal and fitness, in part through Rho kinase activity. Spred1 deficiency mitigates HSC failure induced by infection mimetics and prolongs HSC lifespan, but it does not initiate leukemogenesis due to compensatory upregulation of Spred2. In contrast, HFD induces ERK hyperactivation and aberrant self-renewal in Spred1-deficient HSCs, resulting in functional HSC failure, severe anemia, and myeloproliferative neoplasm-like disease. HFD-induced hematopoietic abnormalities are mediated partly through alterations to the gut microbiota. Together, these findings reveal that diet-induced stress disrupts fine-tuning of Spred1-mediated signals to govern HSC homeostasis.
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http://dx.doi.org/10.1016/j.stem.2018.04.002DOI Listing
May 2018

Identification of antipsychotic drug fluspirilene as a potential anti-glioma stem cell drug.

Oncotarget 2017 Dec 4;8(67):111728-111741. Epub 2017 Dec 4.

Department of Neurosurgery, Graduate School of Medical Science, Kanazawa University, Kanazawa, Japan.

Glioma stem cell (GSC)-targeted therapy is expected to be one of the most innovative approaches to treat patients with glioblastoma (GBM). A number of the drugs that restrain the signaling pathway essential for GSC maintenance have been under clinical trials. Here, we identified fluspirilene, a traditional antipsychotic drug, as a GSC-targeting agent, selected from thousands of existing drugs, and investigated its therapeutic effects against GBM with the purpose of drug repositioning. To develop novel therapeutics targeting GSCs, we initially screened drug libraries for small-molecule compounds showing a greater efficacy, compared to that of controls, in inhibiting the proliferation and survival of different GSC lines using cell proliferation assay. Drugs already reported to show therapeutic effects against GBM or those under clinical trials were excluded from subsequent screening. Finally, we found three drugs showing remarkable antiproliferative effects on GSCs at low concentrations and investigated their therapeutic effects on GSCs, glioma cell lines, and in a GBM mouse model. Of the three compounds, fluspirilene demonstrated a significant inhibitory effect on the proliferation and invasion of glioma cells as well as in the model mice treated with the drug. These effects were associated with the inactivation of the signal transducer and activator of transcription 3 (STAT3). Redeveloping of fluspirilene is a promising approach for the treatment of GBM.
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http://dx.doi.org/10.18632/oncotarget.22904DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5762355PMC
December 2017

Clinical Severity Classification using Automated Conjunctival Hyperemia Analysis Software in Patients with Superior Limbic Keratoconjunctivitis.

Curr Eye Res 2018 06 16;43(6):679-682. Epub 2018 Jan 16.

a Division of Ophthalmology, Department of Visual Sciences , Nihon University School of Medicine , Tokyo , Japan.

Purpose: Digitization of clinical observation is necessary for assessing the severity of superior limbic keratoconjunctivitis (SLK). This study aimed to use a novel quantitative marker to examine hyperemia in patients with SLK.

Materials And Methods: We included six eyes of six patients with both dry eye disease and SLK (SLK group) and eight eyes of eight patients with Sjögren syndrome (SS group). We simultaneously obtained the objective finding scores by using slit-lamp examination and calculated the superior hyperemia index (SHI) with an automated conjunctival hyperemia analysis software by using photographs of the anterior segment. Three objective finding scores, including papillary formation of the superior palpebral conjunctiva, superior limbal hyperemia and swelling, and superior corneal epitheliopathy, were determined. The SHI was calculated as the superior/temporal ratio of bulbar conjunctival hyperemia by using the software. Fisher's exact test was used to compare a high SHI (≥1.07) ratio between the SLK and SS groups. P-Values < 0.05 were considered statistically significant.

Results: The SHI (mean ± standard deviation) in the SLK and SS groups was 1.19 ± 0.50 and 0.69 ± 0.24, respectively. The number of patients with a high SHI (≥1.07) was significantly higher in the SLK group than in the SS group (p < 0.05). The sensitivity and specificity of the SHI in the differential diagnosis between SS and SLK were 66.7% and 87.5%, respectively. An analysis of the association between the objective finding scores and SHI showed that the SHI had a tendency to indicate the severity of superior limbal hyperemia and swelling score in the SLK group.

Conclusion: The SHI calculated using the automated conjunctival hyperemia analysis software could successfully quantify superior bulbar conjunctival hyperemia and may be a useful tool for the differential diagnosis between SS and SLK and for the quantitative follow-up of patients with SLK.
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http://dx.doi.org/10.1080/02713683.2017.1419573DOI Listing
June 2018

Distinct roles of Rheb and Raptor in activating mTOR complex 1 for the self-renewal of hematopoietic stem cells.

Biochem Biophys Res Commun 2018 01 22;495(1):1129-1135. Epub 2017 Nov 22.

Division of Molecular Genetics, Cancer and Stem Cell Research Program, Cancer Research Institute, Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa 920-1192, Japan; WPI Nano Life Science Institute (WPI-Nano LSI), Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa 920-1192, Japan. Electronic address:

The mammalian target of rapamycin (mTOR) complex 1 (mTORC1) senses a cell's energy status and environmental levels of nutrients and growth factors. In response, mTORC1 mediates signaling that controls protein translation and cellular metabolism. Although mTORC1 plays a critical role in hematopoiesis, it remains unclear which upstream stimuli regulate mTORC1 activity in the context of hematopoietic stem cells (HSC) maintenance in vivo. In this study, we investigated the function of Rheb, a critical regulator of mTORC1 activity controlled by the PI3K-AKT-TSC axis, both in HSC maintenance in mice at steady-state and in HSC-derived hematopoiesis post-transplantation. In contrast to the severe hematopoietic dysfunction caused by Raptor deletion, which completely inactivates mTORC1, Rheb deficiency in adult mice did not show remarkable hematopoietic failure. Lack of Rheb caused abnormalities in myeloid cells but did not have impact on hematopoietic regeneration in mice subjected to injury by irradiation. As previously reported, Rheb deficiency resulted in defective HSC-derived hematopoiesis post-transplantation. However, while Raptor is essential for HSC competitiveness in vivo, Rheb is dispensable for HSC maintenance under physiological conditions, indicating that the PI3K-AKT-TSC pathway does not contribute to mTORC1 activity for sustaining HSC self-renewal activity at steady-state. Thus, the various regulatory elements that impinge upstream of mTORC1 activation pathways are differentially required for HSC homeostasis in vivo.
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http://dx.doi.org/10.1016/j.bbrc.2017.11.140DOI Listing
January 2018

Medial rather than lateral knee instability correlates with inferior patient satisfaction and knee function after total knee arthroplasty.

Knee 2017 Dec 29;24(6):1478-1484. Epub 2017 Sep 29.

Department of Orthopaedic Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan.

Background: It is commonly thought that balanced medial and lateral tibiofemoral joint gaps are essential, but the effect of joint laxity on clinical outcome after total knee arthroplasty (TKA) is unclear. It was hypothesised that medial joint laxity correlates with inferior patient satisfaction and knee function, although lateral joint laxity is allowed to a certain degree in TKA.

Methods: This study included 50 knees that underwent primary TKA. Knee laxity was measured with postoperative stress radiographs in flexion and extension, and patient satisfaction and knee function were evaluated by the 2011 Knee Society Knee Scoring System.

Results: In a comparison of medially tight and medially loose knees in flexion, the scores for satisfaction, symptoms, standard activity, and advanced activity were significantly better in medially tight than in medially loose knees (satisfaction: 29.8, 22.2; symptoms: 20.3, 15.9; standard activities: 24.2, 19.1; and advanced activities: 15.3, 8.7, in the tight and loose knees, respectively). Neither lateral joint laxity during knee flexion nor medial joint laxity during knee extension was associated with a poor postoperative clinical outcome, whereas lateral joint laxity and the standard activity score in extension had a moderate positive correlation.

Conclusions: Knees with medial joint laxity during flexion resulted in an inferior postoperative outcome, and lateral joint laxity did not influence patient satisfaction or function. Care should be taken to maintain medial joint stability during the TKA procedure.
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http://dx.doi.org/10.1016/j.knee.2017.09.004DOI Listing
December 2017

Functional dissection of hematopoietic stem cell populations with a stemness-monitoring system based on NS-GFP transgene expression.

Sci Rep 2017 09 12;7(1):11442. Epub 2017 Sep 12.

Division of Molecular Genetics, Cancer Research Institute, Kanazawa University, Kanazawa, Japan.

Hematopoietic stem cells (HSCs) in a steady state can be efficiently purified by selecting for a combination of several cell surface markers; however, such markers do not consistently reflect HSC activity. In this study, we successfully enriched HSCs with a unique stemness-monitoring system using a transgenic mouse in which green florescence protein (GFP) is driven by the promoter/enhancer region of the nucleostemin (NS) gene. We found that the phenotypically defined long-term (LT)-HSC population exhibited the highest level of NS-GFP intensity, whereas NS-GFP intensity was strongly downregulated during differentiation in vitro and in vivo. Within the LT-HSC population, NS-GFP cells exhibited significantly higher repopulating capacity than NS-GFP cells. Gene expression analysis revealed that nine genes, including Vwf and Cdkn1c (p57), are highly expressed in NS-GFP cells and may represent a signature of HSCs, i.e., a stemness signature. When LT-HSCs suffered from remarkable stress, such as transplantation or irradiation, NS-GFP intensity was downregulated. Finally, we found that high levels of NS-GFP identified HSC-like cells even among CD34 cells, which have been considered progenitor cells without long-term reconstitution ability. Thus, high NS-GFP expression represents stem cell characteristics in hematopoietic cells, making this system useful for identifying previously uncharacterized HSCs.
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http://dx.doi.org/10.1038/s41598-017-11909-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5596002PMC
September 2017

Pathohistological investigation of osteochondral tissue obtained during total knee arthroplasty after osteochondral autologous transfer: a case report.

BMC Res Notes 2017 Jun 6;10(1):194. Epub 2017 Jun 6.

Department of Orthopedic Surgery, Knee/Shoulder Surgery & Sports Medicine, Kyoto Shimogamo Hospital, 17 Higashimorigamae-cho, Shimogamo, Sakyo-ku, Kyoto, 606-0866, Japan.

Background: Osteochondral autologous transfer is one of the repair techniques for cartilage defects of knee with promising knee function recovery. There are no reports including histopathological images concerning human osteochondral tissue after osteochondral autologous transfer. This is the first report to present pathohistological findings of transplanted plugs and host tissues extracted from the human body 3 years after osteochondral autologous transfer. This study aimed to explore the cause factor of chronic pain using histological techniques.

Case Presentation: A 67-year-old Japanese man presented with adjusted total knee arthroplasty 3 years after osteochondral autologous transfer. Although in pain, arthroscopic assessment was not severe. The specimens which was gained during total knee arthroplasty were investigated in gross and microscopically using immunohistochemical staining technic. Histological examination revealed that the gap between grafted plugs and host osteochondral tissues was filled with fibrous tissue that stained positive for type I collagen. A degenerative change and some neovascularity were observed in the regenerated tissue and host trabecular bone. Furthermore, cysts and bone marrow edema were observed.

Conclusion: Our data suggests that the host osteochondral morbidity around grafted plugs might be related to chronical pain and revision surgery.
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http://dx.doi.org/10.1186/s13104-017-2513-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5461697PMC
June 2017

Biparietal Thinning: Accidental Death by a Fall from Standing Height.

J Forensic Sci 2017 Sep 7;62(5):1406-1409. Epub 2017 Feb 7.

John A. Burns School of Medicine, University of Hawaii, 651 Ilano St., Honolulu, HI, 96813.

Biparietal thinning resulting in bilateral and symmetrical resorption and loss of thickness of the parietal bones is an uncommon to rare condition in the anthropological and clinical literature. This enigmatic condition of unknown etiology was first reported in the 18th century and has been variously described as a nonmetric trait, anatomical variant, anomaly, and pathology. Biparietal thinning presents grossly and radiographically as oval-shaped depressions in 0.25-0.8% and 0.4-0.5% of individuals and with a higher frequency in females over the age of 60 years. A review of the literature revealed only one example of cranial trauma associated with biparietal thinning and none of fatal trauma associated with this condition. This case reports a rare example of fatal trauma in an elderly man that resulted from a backward fall from a standing height and highlights the increased risk of craniocerebral trauma in individuals with this condition.
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http://dx.doi.org/10.1111/1556-4029.13425DOI Listing
September 2017

An anatomical investigation of clock face landmarks around the glenoid for shoulder arthroscopy orientation.

J Orthop Sci 2016 Nov 31;21(6):727-731. Epub 2016 Aug 31.

Department of Orthopaedic Surgery, Graduate School of Medicine, Kyoto University, 54, Kawahara-cho, Shogoin, Sakyo-ku, Kyoto City, Kyoto Pref., 606-8507, Japan.

Background: For shoulder arthroscopy, few anatomical landmarks are available and inexperienced surgeons tend to be adrift due to the limited visual field of the scope. The purpose of this study was to demonstrate the useful landmarks around the glenoid for accurate orientation, and also the safe distance to avoid suprascapular nerve injury during surgical procedures around the glenoid.

Methods: In 15 human solution-fixed cadavers, a cross-section of the shoulder joint on the labrum surface was created. The positions of the principal anatomical structures surrounding the glenoid were marked on the labrum and measured using our clock face indication system. In 9 shoulders the distances from the labral surface to the spinoglenoid notch were recorded. As an indicator of the scapula size, the distances between the superior and inferior angles of the scapula were also measured.

Results: The average landmark positions in the right shoulder were as follows: center of the attachment of the long tendon 11:59, anterior edge of the supraspinatus 11:59, posterior edge of the base of the coracoid process 12:13, superior edge of the subscapularis 1:03, anterior edge of the base of the coracoid process 1:25, inferior edge of the subscapularis 5:27, inferior edge of the teres minor 6:21, border of the infraspinatus and teres minor 7:43, center of the scapula spine 10:06, border of the supra and infraspinatus 10:27. The average distance from the labral surface to the spinoglenoid notch was 23.17 mm, and that from the superior to inferior angle was 144.93 mm. The Pearson correlation coefficient for these distances was 0.007.

Conclusions: The locations of anatomical landmarks surrounding the glenoid were reliably demonstrated using our clock face indication system. The expected distance from the labral surface to the suprascapular nerve was approximately 23 mm, irrespective of the size of the scapula.
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http://dx.doi.org/10.1016/j.jos.2016.06.017DOI Listing
November 2016

Development of a Fiber-Optic Optical Coherence Tomography Probe for Intraocular Use.

Invest Ophthalmol Vis Sci 2016 07;57(9):OCT568-74

Department of Electrical Engineering and Computer Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi, Japan.

Purpose: To evaluate the performance of a newly developed 23-G optical coherence tomography (OCT) probe in animal and human eyes.

Methods: The probe is a side-imaging OCT device with a scanning beam set 43° to the optical axis and a working distance of 1.5 to 2.0 mm. The performance of the OCT probe was tested during vitrectomy in porcine cadaver eyes and rabbit eyes in situ. Optical coherence tomography images of a normal retina, retinal break, optic disc, pars plicata of the ciliary body, and intraoperative surgical manipulations were recorded. The probe was also tested in a pilot study of clinical cases; intraoperative real-time OCT imaging was performed in three patients, including a 56-year-old woman with an epiretinal membrane.

Results: The OCT probe was able to delineate intraocular tissues, including the posterior retina, and even the most peripheral pars plicata in animal eyes. The OCT probe also successfully delineated intraoperative surgical maneuvers such as membrane peeling and the minute structures of the vortex veins, ora serrata, and vitreous incarceration in the scleral incision from the trocar with sufficient resolution in the patients. There were no complications resulting from its use.

Conclusions: The ability of this new 23-G OCT probe to obtain images of intraoperative manipulations from the most peripheral tissues in animal and patient eyes suggests that it could enable surgeons to make better decisions during the course of intraocular surgery.
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http://dx.doi.org/10.1167/iovs.15-18853DOI Listing
July 2016