Publications by authors named "Masato Saito"

98 Publications

Radiological changes in the formation of Brodie's abscess by sequential magnetic resonance imaging: a case report.

Radiol Case Rep 2021 Oct 6;16(10):2993-2997. Epub 2021 Aug 6.

Department of Orthopaedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan.

Brodie's abscess is a relatively rare subacute form of osteomyelitis. Early diagnosis is challenging because of its insidious onset and vague symptoms. Magnetic resonance imaging is helpful in the diagnosis of Brodie's abscess; however, to date, no study has described the imaging findings of this disease in the early stage. Here, we present the case of a 14 year-old boy with Brodie's abscess in the proximal tibia. The lesion initially presented as a bone marrow edema in the proximal metaphysis of the left tibia on MRI and was misinterpreted as a bone bruise. Further radiological examination was performed 1 month later; this revealed the formation of an abscess cavity, which suggested Brodie's abscess. The patient was referred to our hospital and underwent curettage and debridement, which led to the definitive diagnosis of Brodie's abscess on histopathological findings and bacterial culture. On careful retrospective evaluation, the initial radiological findings suggested a microabscess on the metaphyseal side of the growth plate and bone marrow edema spreading from the lesion to the epiphysis. These radiological changes could be reliable evidence proving that the metaphyseal side of the growth plate is the origin of Brodie's abscess. Moreover, bone marrow edema with suspected microabscess in the metaphysis of the long bones can be the initial stage of the formation of Brodie's abscess and should be carefully followed up.
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http://dx.doi.org/10.1016/j.radcr.2021.07.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8353412PMC
October 2021

A design and optimization of a high throughput valve based microfluidic device for single cell compartmentalization and analysis.

Sci Rep 2021 Jun 21;11(1):12995. Epub 2021 Jun 21.

Graduate School of Engineering, Osaka University, Suita, Osaka, 565-0871, Japan.

The need for high throughput single cell screening platforms has been increasing with advancements in genomics and proteomics to identify heterogeneity, unique cell subsets or super mutants from thousands of cells within a population. For real-time monitoring of enzyme kinetics and protein expression profiling, valve-based microfluidics or pneumatic valving that can compartmentalize single cells is advantageous by providing on-demand fluid exchange capability for several steps in assay protocol and on-chip culturing. However, this technique is throughput limited by the number of compartments in the array. Thus, one big challenge lies in increasing the number of microvalves to several thousand that can be actuated in the microfluidic device to confine enzymes and substrates in picoliter volumes. This work explores the design and optimizations done on a microfluidic platform to achieve high-throughput single cell compartmentalization as applied to single-cell enzymatic assay for protein expression quantification. Design modeling through COMSOL Multiphysics was utilized to determine the circular microvalve's optimized parameters, which can close thousands of microchambers in an array at lower sealing pressure. Multiphysical modeling results demonstrated the relationships of geometry, valve dimensions, and sealing pressure, which were applied in the fabrication of a microfluidic device comprising of up to 5000 hydrodynamic traps and corresponding microvalves. Comparing the effects of geometry, actuation media and fabrication technique, a sealing pressure as low as 0.04 MPa was achieved. Applying to single cell enzymatic assay, variations in granzyme B activity in Jurkat and human PBMC cells were observed. Improvement in the microfluidic chip's throughput is significant in single cell analysis applications, especially in drug discovery and treatment personalization.
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http://dx.doi.org/10.1038/s41598-021-92472-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8217553PMC
June 2021

Gene co-expression network analysis identifies BEH3 as a stabilizer of secondary vascular development in Arabidopsis.

Plant Cell 2021 Aug;33(8):2618-2636

Department of Biology, Graduate School of Science, Kobe University, 1-1 Rokkodai, Kobe 657-8501, Japan.

In plants, vascular stem cells located in the cambium continuously undergo self-renewal and differentiation during secondary growth. Recent advancements in cell sorting techniques have enabled access to the transcriptional regulatory framework of cambial cells. However, mechanisms underlying the robust control of vascular stem cells remain unclear. Here, we identified a new cambium-related regulatory module through co-expression network analysis using multiple transcriptome datasets obtained from an ectopic vascular cell transdifferentiation system using Arabidopsis cotyledons, Vascular cell Induction culture System Using Arabidopsis Leaves (VISUAL). The cambium gene list included a gene encoding the transcription factor BES1/BZR1 Homolog 3 (BEH3), whose homolog BES1 negatively affects vascular stem cell maintenance. Interestingly, null beh3 mutant alleles showed a large variation in their vascular size, indicating that BEH3 functions as a stabilizer of vascular stem cells. Genetic analysis revealed that BEH3 and BES1 perform opposite functions in the regulation of vascular stem cells and the differentiation of vascular cells in the context of the VISUAL system. At the biochemical level, BEH3 showed weak transcriptional repressor activity and functioned antagonistically to other BES/BZR members by competing for binding to the brassinosteroid response element. Furthermore, mathematical modeling suggested that the competitive relationship between BES/BZR homologs leads to the robust regulation of vascular stem cells.
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http://dx.doi.org/10.1093/plcell/koab151DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8408481PMC
August 2021

-Ammonium Ylide Mediators for Electrochemical C-H Oxidation.

J Am Chem Soc 2021 May 13;143(20):7859-7867. Epub 2021 May 13.

Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, United States.

The site-specific oxidation of strong C(sp)-H bonds is of uncontested utility in organic synthesis. From simplifying access to metabolites and late-stage diversification of lead compounds to truncating retrosynthetic plans, there is a growing need for new reagents and methods for achieving such a transformation in both academic and industrial circles. One main drawback of current chemical reagents is the lack of diversity with regard to structure and reactivity that prevents a combinatorial approach for rapid screening to be employed. In that regard, directed evolution still holds the greatest promise for achieving complex C-H oxidations in a variety of complex settings. Herein we present a rationally designed platform that provides a step toward this challenge using -ammonium ylides as electrochemically driven oxidants for site-specific, chemoselective C(sp)-H oxidation. By taking a first-principles approach guided by computation, these new mediators were identified and rapidly expanded into a library using ubiquitous building blocks and trivial synthesis techniques. The ylide-based approach to C-H oxidation exhibits tunable selectivity that is often exclusive to this class of oxidants and can be applied to real-world problems in the agricultural and pharmaceutical sectors.
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http://dx.doi.org/10.1021/jacs.1c03780DOI Listing
May 2021

Deskilled and Rapid Drug-Resistant Gene Detection by Centrifugal Force-Assisted Thermal Convection PCR Device.

Sensors (Basel) 2021 Feb 9;21(4). Epub 2021 Feb 9.

Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.

Here we report the improved Cyclo olefin polymer (COP) microfluidic chip and polymerase chain reaction (PCR) amplification system for point-of-care testing (POCT) in rapid detection of Carbapenem-resistant Enterobacteriaceae (CRE). The PCR solution and thermal cycling is controlled by the relative gravitational acceleration (7G) only and is expected to pose minimal problem in operation by non-expert users. Detection is based on identifying the presence of carbapenemase encoding gene through the corresponding fluorescence signal after amplification. For preliminary tests, the device has been demonstrated to detect from patients stool samples. From the prepared samples, 96.4 fg/µL was detected with good certainty within 15 min (~106 thermocycles,) which is significantly faster than the conventional culture plate method. Moreover, the device is expected to detect other target genes in parallel as determination of the presence of and from control samples has also been demonstrated. With the rising threat of drug-resistant bacteria in global healthcare, this technology can greatly aid the health sector by enabling the appropriate use of antibiotics, accelerating the treatment of carriers, and suppressing the spread.
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http://dx.doi.org/10.3390/s21041225DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7916093PMC
February 2021

Enhancement of Skin Permeation of a Hydrophilic Drug from Acryl-Based Pressure-Sensitive Adhesive Tape.

Pharm Res 2021 Feb 29;38(2):289-299. Epub 2021 Jan 29.

Graduate School of Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado, Saitama, 350-0295, Japan.

Purpose: Penetration enhancers are necessary to overcome a formidable barrier function of the stratum corneum in the development of topical formulations. Recently, non-lamella liquid crystal (NLLC)-forming lipids such as glycerol monooleate and phytantriol (PHY) are gaining increasing attention as a novel skin permeation enhancer. In the present study, fluorescein sodium (FL-Na) was used as a model hydrophilic drug, and acryl-base pressure-sensitive adhesive (PSA) tape containing NLLC forming lipids, mono-O-(5,9,13-trimethyl-4-tetradecenyl) glycerol ester (MGE) or PHY, was prepared to enhance drug permeation through the skin.

Methods: A PSA patch containing FL-Na was prepared by mixing FL-Na entrapped in NLLC and acrylic polymer. FL permeation through excised hairless rat skin, and also human skin, was investigated. Changes in lipid structure, folding/unfolding state of keratin in the stratum corneum, and penetration of MGE into the stratum corneum were investigated using confocal Raman microscopy.

Results: Enhanced FL permeation was observed by the application of a PSA patch containing MGE and PHY. Especially, dramatically enhancement effect was confirmed by 15% of MGE contained formulation. Penetration of MGE provided diminished orthorhombic crystal structure and a peak shift of the aliphatic CH vibration of keratin chains toward lower wavenumbers.

Conclusion: The present results suggested that the formulation development by adding MGE may be useful for improving the skin permeation of mal-permeable drugs such as hydrophilic drugs.
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http://dx.doi.org/10.1007/s11095-021-02996-zDOI Listing
February 2021

Temperature-dependent fasciation mutants provide a link between mitochondrial RNA processing and lateral root morphogenesis.

Elife 2021 Jan 14;10. Epub 2021 Jan 14.

Botanical Gardens, Graduate School of Science, The University of Tokyo, Tokyo, Japan.

Although mechanisms that activate organogenesis in plants are well established, much less is known about the subsequent fine-tuning of cell proliferation, which is crucial for creating properly structured and sized organs. Here we show, through analysis of temperature-dependent fasciation (TDF) mutants of Arabidopsis, (), , and (), that mitochondrial RNA processing is required for limiting cell division during early lateral root (LR) organogenesis. These mutants formed abnormally broadened (i.e. fasciated) LRs under high-temperature conditions due to extra cell division. All TDF proteins localized to mitochondria, where they were found to participate in RNA processing: RRD1 in mRNA deadenylation, and RRD2 and RID4 in mRNA editing. Further analysis suggested that LR fasciation in the TDF mutants is triggered by reactive oxygen species generation caused by defective mitochondrial respiration. Our findings provide novel clues for the physiological significance of mitochondrial activities in plant organogenesis.
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http://dx.doi.org/10.7554/eLife.61611DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7846275PMC
January 2021

Common iliac artery dissection as a complication of common iliac artery balloon occlusion for placenta percreta: A case report.

J Obstet Gynaecol Res 2021 Mar 14;47(3):1172-1177. Epub 2020 Dec 14.

Department of Obstetrics and Gynecology, Sapporo Medical University School of Medicine, Sapporo, Japan.

A 37-year-old pregnant woman who had undergone three previous cesarean sections was diagnosed as having placenta percreta. We decided to perform cesarean hysterectomy with bilateral common iliac artery balloon occlusion (CIABO). The duration of surgery was 2 h and 2 min and total estimated blood loss was 2600 mL. Surgery was completed without any surgical complications, but the pulse oximeter waveform of the left leg became undetectable during surgery. We immediately performed angiography after closure of laparotomy and found abnormal pooling of contrast media at the left common iliac artery in the region in which the balloon was positioned. We made a diagnosis of left common iliac artery dissection caused by CIABO. We performed emergent revascularization by intravascular stenting. We conclude that CIABO can cause common iliac artery dissection by mechanical stimulation of the inflated balloon. Careful intraoperative evaluation of limb ischemia and preparation of intravascular treatment is needed for a safe procedure.
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http://dx.doi.org/10.1111/jog.14601DOI Listing
March 2021

[Lactic Acidosis after Cardiac Surgery Treated by Continuous Renal Replacement Therapy in a Patient on Metformin;Report of a Case].

Kyobu Geka 2020 Dec;73(13):1121-1123

Department of Cardiovascular Surgery, Sendai City Medical Center, Sendai, Japan.

Lactic acidosis is a common condition observed in a patient after cardiac surgery. It is important to identify the pathogenesis of lactic acidosis since the delay of an appropriate treatment leads to high mortality. Metformin overdose has been known as a risk of lactic acidosis, and previous reports have demonstrated that continuous renal replacement therapy(CRRT) is effective. However, it has yet to be known if metformin-related lactic acidosis after cardiac surgery is treatable with CRRT. We, herein, report a case of 64-year-old diabetic male who had been on metformin treatment until 1 day before surgery. He presented lactic acidosis postoperatively and was successfully treated with CRRT. This case suggests that it is necessary to discontinue metformin no later than 2 days before surgery and that CRRT is of use for perioperative lactic acidosis in a patient on metformin.
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December 2020

study of monocytic THP-1 leukemia cell membrane elasticity with a single-cell microfluidic-assisted optical trapping system.

Biomed Opt Express 2020 Oct 1;11(10):6027-6037. Epub 2020 Oct 1.

OPTICS Research Unit, CENSER, De La Salle University (DLSU), Manila, Philippines.

We studied the elastic profile of monocytic THP-1 leukemia cells using a microfluidic-assisted optical trap. A 2-m fused silica bead was optically trapped to mechanically dent an immobilized single THP-1 monocyte sieved on a 15-µm microfluidic capture chamber. Cells treated with Zeocin and untreated cells underwent RT-qPCR analysis to determine cell apoptosis through gene expression in relation to each cell's deformation profile. Results showed that untreated cells with 43.05 ± 6.68 Pa are more elastic compared to the treated cells with 15.81 ± 2.94 Pa. THP-1 monocyte's elastic modulus is indicative of cell apoptosis shown by upregulated genes after Zeocin treatment. This study clearly showed that the developed technique can be used to distinguish between cells undergoing apoptosis and cells not undergoing apoptosis and which may apply to the study of other cells and other cell states as well.
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http://dx.doi.org/10.1364/BOE.402526DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7587289PMC
October 2020

Clinical Outcomes of Patients with Metastatic Solitary Fibrous Tumors: A Japanese Musculoskeletal Oncology Group (JMOG) Multiinstitutional Study.

Ann Surg Oncol 2021 Jul 4;28(7):3893-3901. Epub 2020 Nov 4.

Department of Orthopedic Surgery, National Hospital Organization Osaka National Hospital, Osaka, Japan.

Background: Although the unpredictable malignant behavior of solitary fibrous tumors (SFTs) has been recognized, the clinical features and prognosis of metastatic SFTs have not been well documented due to the extreme rarity of these cases. The aim of this study is to investigate the clinical features, prognostic factors, and optimal management of patients with metastatic SFTs.

Patients And Methods: Sixty patients with metastatic SFT were retrospectively reviewed. Univariate and multivariate analyses were performed to identify the factors associated with survival. Time to next treatment (TNT) was used to evaluate the effects of various chemotherapy regimens.

Results: A total of 34 male and 26 female patients (median age 55 years, range, 23-87 years) were included in the study. The median follow-up period after metastasis was 32 months (range 1-126 months). Tumor location and local recurrence were correlated with late metastasis. The 3- and 5-year overall survival rates were 72.7% and 49.2%, respectively. Primary tumor location, number of metastases, and metastasectomy were significantly associated with survival. Metastasectomy was the only significant variable on multivariate analysis. The TNT was significantly different among the various regimens.

Conclusions: Patients with metastatic SFTs had relatively longer survival periods compared with those with other metastatic soft-tissue sarcomas. Tumor location and number of metastases was associated with survival. Surgical resection of the metastatic lesions offers the best chance of survival, however further studies are warranted to define patients who would benefit from metastasectomy, and the most effective chemotherapeutic regimen for patients with metastatic SFTs remains unknown.
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http://dx.doi.org/10.1245/s10434-020-09306-8DOI Listing
July 2021

The future of microfluidics in immune checkpoint blockade.

Cancer Gene Ther 2021 Sep 27;28(9):895-910. Epub 2020 Oct 27.

Graduate School of Engineering, Osaka University, Suita, Osaka, 565-0871, Japan.

Recent advances in microfluidic techniques have enabled researchers to study sensitivities to immune checkpoint therapy, to determine patients' response to particular antibody treatment. Utilization of this technology is helpful in antibody discovery and in the design of personalized medicine. A variety of microfluidic approaches can provide several functions in processes such as immunologic, genomic, and/or transcriptomic analysis with the aim of improving the efficacy and coverage of immunotherapy, particularly immune checkpoint blockade (ICB). To achieve this requires researchers to overcome the challenges in the current state of the technology. This review looks into the advancements in microfluidic technologies applied to researches on immune checkpoint blockade treatment and its potential shift from proof-of-principle stage to clinical application.
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http://dx.doi.org/10.1038/s41417-020-00248-7DOI Listing
September 2021

Early recurrence of Rosai-Doefman disease after total removal resection: a case report.

Br J Neurosurg 2020 Sep 16:1-3. Epub 2020 Sep 16.

Neurosurgery, Asahikawa Medical University, Asahikawa, Japan.

Background: Rosai-Dorfman disease (RDD) involving the central nervous system (CNS) is rare and observed in 5% of all patients with extranodal RDD. According to a previous report, gross total resection is curative; however, we encountered a case of recurrence following gross total resection. We discuss our case and review previous reports on recurrent RDD.

Case Description: A 68-year-old woman came to the hospital complaining of left parietal mass. A tumor that had partially eroded the frontal bone was found. As the lesion was suspected to be malignant, we performed a total resection. Pathology results were indicative of an RDD. We did not prescribe adjuvant therapy because total resection was performed. However, after a year, abnormal accumulation in the left parietal bone was observed on FDG-PET. This was considered as recurrence, and re-excision was performed. Pathological assessments confirmed the recurrence of RDD.

Conclusions: Our case demonstrated the recurrence of RDD following total resection. Future reports should assess these peculiarities. This will facilitate discussions on the risk factors and the effectiveness of treatment methods.
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http://dx.doi.org/10.1080/02688697.2020.1817316DOI Listing
September 2020

Arteriovenous malformation in pancreas mimicking hypervascular tumor.

JGH Open 2020 Aug 18;4(4):773-774. Epub 2020 Apr 18.

Department of Gastroenterology and Hepatology Sapporo Medical University School of Medicine Sapporo Japan.

Arteriovenous malformation (AVM) is defined as a disease that causes blood flow abnormality due to anastomoses of the arteries and veins. AVM can occur in any gastrointestinal tract, but pancreatic AVM (P-AVM) is very rare. Previous reports demonstrated that contrast-enhanced CT (CECT) typically showed abnormal vascular network in pancreas. We present a 58-year old man with a history of acute pancreatitis. He was referred to our hospital for examination of pancreatic mass. CECT showed a round-shaped hypervascular lesion with a diameter of 8 mm in the head of the pancreas. Selective angiography showed vascular network and early visualization of superior mesenteric vein. We finally diagnosed this case as P-AVM. He underwent duodenum preserving pancreatic head resection. Histological findings confirmed the preoperative diagnosis of P-AVM.
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http://dx.doi.org/10.1002/jgh3.12343DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7411645PMC
August 2020

Towards On-site Determination of Secretory IgA in Artificial Saliva with Gold-Linked Electrochemical Immunoassay (GLEIA) Using Portable Potentiostat and Disposable Printed Electrode.

Appl Biochem Biotechnol 2021 May 13;193(5):1311-1320. Epub 2020 Jun 13.

AIST-Osaka University Advanced Photonics and Biosensing Open Innovation Laboratory, AIST, Suita, Japan.

Mental stress is closely connected with our physical and mental wellness. Therefore, stress measurement can contribute to assess our lifestyle and increase our quality of life. In this paper, we detect the secretory immunoglobulin A (sIgA), which is the candidate of salivary stress markers, with original electrochemical immunoassay: gold-linked electrochemical immunoassay (GLEIA). This biosensor is based on a sandwich-type immunosensor and adopts the electrochemical method to detect the reduction peak from Au nanoparticles linked to the secondary antibody. GLEIA is convenient and cost-effective that only requires a low sample volume (10 μL). In addition, the GLEIA show high sensitivity and selectivity. We obtained the linear response to relate the concentration of sIgA (10-300 ng/mL) in D-PBS buffer with the artificial saliva which includes salivary inorganic salt and typically glycoprotein (mucin). Furthermore, we obtained acceptable selectivity in the various solution with salivary proteins such as α-amylase, human serum albumin, immunoglobulin G (IgG), lysozyme, and mucin. In the future, we try to detect the sIgA in real saliva for on-site stress measurement using GLEIA and to integrate the various immunosensors for stress markers in saliva.
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http://dx.doi.org/10.1007/s12010-020-03332-8DOI Listing
May 2021

Robust CD8+ T-cell proliferation and diversification after mogamulizumab in patients with adult T-cell leukemia-lymphoma.

Blood Adv 2020 05;4(10):2180-2191

Department of Hematology and Oncology and.

Skin-related adverse events (AEs) occur frequently in adult T-cell leukemia-lymphoma (ATL) patients treated with mogamulizumab, a humanized anti-CCR4 monoclonal antibody. This study was undertaken to elucidate the mechanisms of mogamulizumab-induced skin-related AEs. We analyzed the T-cell receptor β chain repertoire in ATL patients' peripheral blood mononuclear cells (PBMCs) before and after mogamulizumab. Skin-related AEs were present in 16 patients and were absent in 8 patients. Additionally, we included 11 patients before and after chemotherapy without mogamulizumab. Immune-related gene expression in PBMCs before and after mogamulizumab was also assessed (n = 24). Mogamulizumab treatment resulted in CCR4+ T-cell depletion, and the consequent lymphopenia provoked homeostatic CD8+ T-cell proliferation, as evidenced by increased expressions of CD8B and CD8A, which were significantly greater in patients with skin-related AEs than in those without them. We hypothesize that proliferation is driven by the engagement of self-antigens, including skin-related antigens, in the face of regulatory T-cell depletion. Together with the observed activated antigen presentation function, this resulted in T-cell diversification that was significantly greater in patients with skin-related AEs than in those without. We found that the CD8+ T cells that proliferated and diversified after mogamulizumab treatment were almost entirely newly emerged clones. There was an inverse relationship between the degree of CCR4+ T-cell depletion and increased CD8+ T-cell proliferation and diversification. Thus, lymphocyte-depleting mogamulizumab treatment provokes homeostatic CD8+ T-cell proliferation predominantly of newly emerging clones, some of which could have important roles in the pathogenesis of mogamulizumab-induced skin-related AEs.
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http://dx.doi.org/10.1182/bloodadvances.2020001641DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7252545PMC
May 2020

Successful pregnancy and delivery achieved following intracytoplasmic sperm injection using teratozoospermic sperm exhibiting marked dysplasia of the fibrous sheath: a case report.

Transl Androl Urol 2020 Apr;9(2):800-806

Department of Reproductive Medicine, Kameda IVF Clinic Makuhari, Chiba, Japan.

A husband and his wife, both 34 years old, consulted our clinic because of primary infertility. Sperm analysis revealed that the sperm concentration, motility, and progressive motility were (42.8±22.8)×10/mL, 23.3%±12.2%, and 12.9%±6.1%, respectively. Based on Krugar strict morphology criteria, 100% of the sperm were teratozoospermic, with 7.9% DNA fragmentation index. Observation of the sperm under a transmission electron microscope revealed that most parts of the fibrous sheath (FS) surrounding the tails of the sperm were missing from midway through the principal piece to the end piece, although the sperm's heads, necks and midpieces were morphologically normal. To collect oocytes, the gonadotropin-releasing hormone antagonist protocol was carried out, and 7 oocytes were retrieved. Intracytoplasmic sperm injection (ICSI) was performed for all the teratozoospermic sperm. Of the 7 oocytes, 3 were fertilized, and one 8-cell embryo and 2 expanded blastocysts were vitrified. Although repeated transfers of expanded blastocysts resulted in no implantation, one 8-cell embryo transfer in a hormone replacement therapy cycle led to pregnancy. The pregnancy using an 8-cell vitrified embryo resulted in the delivery of a healthy female baby at 38 weeks of gestation. No congenital malformations were found until 28 days after birth. Our results demonstrated that healthy birth could be achieved following the transfer of an embryo derived from ICSI using teratozoospermic sperm exhibiting the dysplasia of the fibrous sheath (DFS). Furthermore, while the previous reports on DFS have not investigated male infertility, we evaluated sperms from various aspects such as Kruger sperm function test, chromatin dispersion test, electron microscopy findings, time-lapse images of the obtained embryos, and concluded that ICSI could be desirable as a treatment policy for DFS.
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http://dx.doi.org/10.21037/tau.2019.12.14DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7215008PMC
April 2020

VISUAL-CC system uncovers the role of GSK3 as an orchestrator of vascular cell type ratio in plants.

Commun Biol 2020 04 22;3(1):184. Epub 2020 Apr 22.

Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, 113-0033, Japan.

The phloem transports photosynthetic assimilates and signalling molecules. It mainly consists of sieve elements (SEs), which act as "highways" for transport, and companion cells (CCs), which serve as "gates" to load/unload cargos. Though SEs and CCs function together, it remains unknown what determines the ratio of SE/CC in the phloem. Here we develop a new culture system for CC differentiation in Arabidopsis named VISUAL-CC, which almost mimics the process of the SE-CC complex formation. Comparative expression analysis in VISUAL-CC reveals that SE and CC differentiation tends to show negative correlation, while total phloem differentiation is unchanged. This varying SE/CC ratio is largely dependent on GSK3 kinase activity. Indeed, gsk3 hextuple mutants possess many more SEs and fewer CCs, whereas gsk3 gain-of-function mutants partially increase the CC number. Taken together, GSK3 activity appears to function as a cell-fate switch in the phloem, thereby balancing the SE/CC ratio.
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http://dx.doi.org/10.1038/s42003-020-0907-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7176705PMC
April 2020

Mild and Chemoselective Thioacylation of Amines Enabled by the Nucleophilic Activation of Elemental Sulfur.

J Am Chem Soc 2020 05 27;142(18):8130-8135. Epub 2020 Apr 27.

Graduate School of Pharmaceutical Sciences, Kyoto University, Yoshida, Sakyo-ku, Kyoto 606-8501, Japan.

A mild and chemoselective method for the thioacylation of amines using α-keto acids and elemental sulfur has been developed. The key to the success of this transformation is the nucleophilic activation of elemental sulfur by thiols such as 1-dodecanethiol. A variety of functional groups, including unprotected hydroxyl, carboxyl, amide, sulfide, and tertiary amine moieties, are tolerated under the applied reaction conditions. To demonstrate the advantages of this method compared with conventional O-S exchange reactions using Lawesson's reagent or PS, thioamide moieties were introduced site-specifically into biologically active compounds.
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http://dx.doi.org/10.1021/jacs.0c03256DOI Listing
May 2020

Real-Time Monitoring and Detection of Single-Cell Level Cytokine Secretion Using LSPR Technology.

Micromachines (Basel) 2020 Jan 19;11(1). Epub 2020 Jan 19.

Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.

Cytokine secretion researches have been a main focus of studies among the scientists in the recent decades for its outstanding contribution to clinical diagnostics. Localized surface plasmon resonance (LSPR) technology is one of the conventional methods utilized to analyze these issues, as it could provide fast, label-free and real-time monitoring of biomolecule binding events. However, numerous LSPR-based biosensors in the past are usually utilized to monitor the average performance of cell groups rather than single cells. Meanwhile, the complicated sensor structures will lead to the fabrication and economic budget problems. Thus, in this paper, we report a simple synergistic integration of the cell trapping of microwell chip and gold-capped nanopillar-structured cyclo-olefin-polymer (COP) film for single cell level Interleukin 6 (IL-6) detection. Here, in-situ cytokine secreted from the trapped cell can be directly observed and analyzed through the peak red-shift in the transmittance spectrum. The fabricated device also shows the potential to conduct the real-time monitoring which would greatly help us identify the viability and biological variation of the tested single cell.
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http://dx.doi.org/10.3390/mi11010107DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7019717PMC
January 2020

Single Cell Analysis of Neutrophils NETs by Microscopic LSPR Imaging System.

Micromachines (Basel) 2019 Dec 31;11(1). Epub 2019 Dec 31.

Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan.

A simple microengraving cell monitoring method for neutrophil extracellular traps (NETs) released from single neutrophils has been realized using a polydimethylsiloxane (PDMS) microwell array (MWA) sheet on a plasmon chip platform. An imbalance between NETs formation and the succeeding degradation (NETosis) are considered associated with autoimmune disease and its pathogenesis. Thus, an alternative platform that can conduct monitoring of this activity on single cell level at minimum cost but with great sensitivity is greatly desired. The developed MWA plasmon chips allow single cell isolation of neutrophils from 150 µL suspension (6.0 × 10 cells/mL) with an efficiency of 36.3%; 105 microwells with single cell condition. To demonstrate the utility of the chip, trapped cells were incubated between 2 to 4 h after introducing with 100 nM phorbol 12-myristate 13-acetate (PMA) before measurement. Under observation using a hyperspectral imaging system that allows high-throughput screening, the neutrophils stimulated by PMA solution show a significant release of fibrils and NETs after 4 h, with observed maximum areas between 314-758 µm. An average absorption peak wavelength shows a redshift of Δλ = 1.5 nm as neutrophils release NETs.
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http://dx.doi.org/10.3390/mi11010052DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7019790PMC
December 2019

A Microfluidic Platform for Single Cell Fluorometric Granzyme B Profiling.

Theranostics 2020 1;10(1):123-132. Epub 2020 Jan 1.

Graduate School of Engineering, Osaka University, Suita, Osaka, 565-0871, JAPAN.

Granzyme B (GrB) is an essential cytotoxic effector in cancer immunotherapy as it can be a potential biomarker to predict the efficacy of immunotherapies including checkpoint inhibitors. Monitoring the Granzyme B activity in cells would help determine a patient's clinical response to treatment and lead to better treatment strategies by preventing administration of ineffective therapies and avoid adverse events resulting in a delay in subsequent treatment. : A microfluidic device with hydrodynamic traps and pneumatic valving system was fabricated using photo and soft lithography. Single cell Granzyme B (GrB) activity was detected and measured fluorometrically using a commercial assay kit with a peptide substrate containing GrB recognition sequence (Ac-IEPD-AFC) and AFC (7-Amino-4-trifluoromethylcoumarin) label. Fluorescence was observed and measured using a confocal microscope with CSU-W1 scanner unit and CCD camera as well as an inverted microscope with photodetector. Model cells (NK-92, GrB-transduced Jurkat, and THP1 cells) and human PBMCs from healthy donor and lung cancer patients including an anti-PD-1 antibody treated patient were profiled of its GrB activity as proof of concept. : GrB expression from the model cells was found to be markedly different. NK-92 cells were found to have higher GrB activity than the GrB-transduced Jurkat cells. THP-1 was found to have relatively no significant activity. A marked increase in GrB expression was also observed in anti-PD-1 treated lung cancer patient sample in comparison to PBMC from a healthy donor. TCR+ Ig-G4+ PBMC cells were found to have high activity which signifies a clear response to PD-1 blockade. : As proof of concept, we have shown the capability of a microfluidic platform to measure GrB production through a single cell enzymatic activity assay. Our platform might be a promising tool for evaluating the sensitivity of immunotherapies and identifying specific T cell subset responsible for the anti-tumor response.
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http://dx.doi.org/10.7150/thno.37728DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6929635PMC
April 2021

Utility of Centrifugation-Controlled Convective (C3) Flow for Rapid On-chip ELISA.

Sci Rep 2019 12 27;9(1):20150. Epub 2019 Dec 27.

Department of Applied Physics, Osaka University, 2-1 Yamadaoka, Suita, 565-0871, Osaka, Japan.

Miniaturizing the enzyme-linked immunosorbent assay (ELISA) protocols in microfluidics is sought after by researchers for a rapid, high throughput screening, on-site diagnosis, and ease in operation for detection and quantification of biomarkers. Herein, we report the use of the centrifugation-controlled convective (C3) flow as an alternative method in fluid flow control in a ring-structured channel for enhanced on-chip ELISA. A system that consists of a rotating heater stage and a microfluidic disk chip has been developed and demonstrated to detect IgA. The ring-structured channel was partially filled with microbeads (250 µm in diameter) carrying the capture antibodies and the analyte solution was driven by thermal convection flow (50 µL/min) to promote the reaction. The remaining part of the circular channel without microbeads served as the observation area to measure the absorbance value of the labeled protein. Currently, the system is capable of conducting four reactions in parallel and can be performed within 30 min at 300 G. A detection limit of 6.16 ng/mL using 24 µL of target sample (IgA) was observed. By simply changing the capture antibodies, the system is expected to be versatile for other immunoassays.
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http://dx.doi.org/10.1038/s41598-019-56772-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6934823PMC
December 2019

Effects of full-length human amelogenin on the differentiation of dental epithelial cells and osteoblastic cells.

Arch Oral Biol 2019 Nov 8;107:104479. Epub 2019 Jul 8.

Department of Pharmacology, School of Dentistry, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido 061-0293, Japan. Electronic address:

Background And Objective: Amelogenins are major components of extracellular matrix proteins in developing teeth, and regulate the growth of enamel crystals. They also function as signaling molecules in cell differentiation. This study aimed to determine the biological effects of amelogenins on the differentiation of HAT-7 dental epithelial cells and MC3T3-E1 pre-osteoblastic cells using full-length recombinant human amelogenin (rh-AMEL).

Design: rh-AMEL was expressed in a mammalian cell line (Expi293F™) and was purified by DDK agarose beads. Effects of rh-AMEL on differentiation were evaluated by Mineralization and Alkaline phosphatase (ALP) activity using Alizarin Red S staining and colorimetric substrate p-nitrophenol, respectively.

Results: Western blotting and silver staining confirmed the successful purification of rh-AMEL. Mineralization and ALP activity in HAT-7 cells were significantly higher after treatment with 4 μg/mL rh-AMEL, but not after treatment with Emdogain (EMD). In MC3T3-E1 cells, on the other hand, rh-AMEL showed biphasic effects on differentiation. Treatment with low concentrations of rh-AMEL (0.001-0.1 μg/mL) and EMD (0.01-1 μg/mL) increased mineralization and ALP activity in MC3T3-E1 cells, whereas treatment with high concentrations of rh-AMEL (4 μg/mL) and EMD (100 μg/mL) had the opposite effect.

Conclusion: High concentrations of rh-AMEL and EMD decreased the differentiation of MC3T3-E1 cells. By contrast, a high concentration of rh-AMEL, but not that of EMD, promoted the differentiation of HAT-7 cells. This study demonstrates that the effects of rh-AMEL on cell differentiation differ between HAT-7 and MC3T3-E1 cells, and suggests that different regions on AMEL may induce the differentiation of these cell types.
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http://dx.doi.org/10.1016/j.archoralbio.2019.07.004DOI Listing
November 2019

Divergent and Chemoselective Transformations of Thioamides with Designed Carbene Equivalents.

Chemistry 2019 Aug 10;25(44):10314-10318. Epub 2019 Jul 10.

Graduate School of Pharmaceutical Sciences, Kyoto University, 46-29 Shimoadachi-cho, Yoshida, Sakyo-ku, Kyoto, 606-8501, Japan.

The reactions of thioamides with ortho-nitro-substituted iodonium ylides proceeded under mild conditions to give enaminones or thiazoles, depending on the iodonium ylide used. This protocol allowed the use of protic solvents, including aqueous solutions, and therefore coupling reactions with complex molecules such as peptides or steroids were possible. A mild and efficient method for the synthesis of various iodonium ylides was established. DFT calculations suggested that the halogen bonding between a thioamide and iodonium ylide was important in this chemoselective coupling reaction. The potential use of enaminones conjugated with pharmaceuticals as prodrugs was also demonstrated.
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http://dx.doi.org/10.1002/chem.201902699DOI Listing
August 2019

What Can Cell Culture Systems Reveal About Sieve Element Differentiation?

Methods Mol Biol 2019 ;2014:459-466

Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan.

Tissue culture systems can be powerful tools for studying the process of cell differentiation in detail. Although a large number of cultures for xylem differentiation have been developed and utilized, there are only few reports on culture systems for ectopic phloem differentiation. Recently, a novel tissue culture system named Vascular cell Induction culture System Using Arabidopsis Leaves (VISUAL) was established, in which both xylem and phloem cells can be rapidly and efficiently induced in the model plant Arabidopsis thaliana. This chapter discusses the principle of VISUAL and how it can be used to investigate phloem differentiation, for example in combination with genetic experiments or transcriptome analysis. In addition, the protocol for establishing a phloem cell culture is provided.
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http://dx.doi.org/10.1007/978-1-4939-9562-2_36DOI Listing
March 2020

Mogamulizumab Treatment Elicits Autoantibodies Attacking the Skin in Patients with Adult T-Cell Leukemia-Lymphoma.

Clin Cancer Res 2019 07 24;25(14):4388-4399. Epub 2019 Apr 24.

Department of Hematology and Oncology, Nagoya City University Graduate School of Medical Sciences, Aichi, Japan.

Purpose: The anti-CCR4 mAb, mogamulizumab, offers therapeutic benefit to patients with adult T-cell leukemia-lymphoma (ATL), but skin-related adverse events (AE) such as erythema multiforme occur frequently. The purpose of this study was to determine the mechanisms by which mogamulizumab causes skin-related AEs in patients with ATL.

Experimental Design: We investigated whether autoantibodies were present in patients' sera using flow cytometry to determine binding to keratinocytes and melanocytes ( = 17), and immunofluorescence analysis of tissue sections. We analyzed the IgM heavy chain repertoire in peripheral blood mononuclear cells before and after mogamulizumab or other chemotherapy by next-generation sequencing (NGS; = 16).

Results: Autoantibodies recognizing human keratinocytes or melanocytes were found in the sera of 6 of 8 patients suffering from mogamulizumab-induced erythema multiforme. In one patient, complement-dependent cytotoxicity (CDC) mediated by autoantibodies against keratinocytes or melanocytes was proportionally related to the severity of the erythema multiforme. The presence of autoantibodies in the epidermis was confirmed in all biopsy specimens of mogamulizumab-induced erythema multiforme ( = 12). Furthermore, colocalization of autoantibodies and C1q, suggesting the activation of CDC, was observed in 67% (8/12). In contrast, no autoantibody or C1q was found in ATL tumor skin lesions ( = 13). Consistent with these findings, NGS demonstrated that IgM germline genes had newly emerged and expanded, resulting in IgM repertoire skewing at the time of erythema multiforme.

Conclusions: Mogamulizumab elicits autoantibodies playing an important role in skin-related AEs, possibly associated with regulatory T-cell depletion. This is the first report demonstrating the presence of skin-directed autoantibodies after mogamulizumab treatment.
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http://dx.doi.org/10.1158/1078-0432.CCR-18-2575DOI Listing
July 2019

Delayed retroperitoneal hemorrhage during veno-venous extracorporeal membrane oxygenation: a case report.

Acute Med Surg 2019 Apr 10;6(2):180-184. Epub 2019 Jan 10.

Department of Emergency Medicine Sapporo Medical University Sapporo Hokkaido Japan.

Case: There are several reports of retroperitoneal hemorrhage induced by percutaneous femoral cannulation for extracorporeal membrane oxygenation (ECMO). However, there are no reports of delayed retroperitoneal hemorrhage, which develops a few days after ECMO initiation and is unrelated to the ECMO cannulation. Herein, we report a rare case of delayed retroperitoneal hemorrhage during veno-venous extracorporeal membrane oxygenation (VV-ECMO).

Outcome: A 54-year-old man was referred to our hospital because of severe acute respiratory distress syndrome. We initiated VV-ECMO. The patient had severe delirium and was confined to a long-term supine position to maintain circuit safety. On day 13, computed tomography unexpectedly revealed a large retroperitoneal hemorrhage spreading around the psoas muscle.

Conclusion: Delayed retroperitoneal hemorrhage can develop during VV-ECMO management a few days after its initiation. Anticoagulant use and forceful muscular strain could be risk factors of delayed retroperitoneal hemorrhage.
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http://dx.doi.org/10.1002/ams2.385DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6442533PMC
April 2019

Treatment of a fourth ventricle arachnoid cyst via anterior hone of lateral ventricle using a flexible endoscope.

World Neurosurg 2019 Jan 21. Epub 2019 Jan 21.

Background: Arachnoid cysts are common anomalies in the intracranial region. However, an intraventricular arachnoid cyst is rare, and occurrence within the fourth ventricle is especially uncommon; only 16 cases have been described in the literature. Arachnoid cysts in the fourth ventricle may cause obstructive hydrocephalus or cerebellar ataxia or cranial nerve palsy. The treatment for a fourth ventricular arachnoid cyst is complete or partial resection via a midline suboccipital approach. Recently, endoscopic fenestration has become the procedure of choice in the treatment of arachnoid cysts in supratentorial locations, but as yet there has been no report of treating a fourth ventricle arachnoid cyst using a flexible endoscope.

Case Description: We present the case of a 43-year-old man, who suffered a recurrence of an arachnoid cyst in the fourth ventricle that had been partially excised 8 years previously using midline suboccipital craniectomy. Because of concerns of adhesions following the previous craniectomy, we decided to perform endoscopic treatment via the anterior horn of the lateral ventricle.

Conclusions: As per our knowledge, this is the first case reporting the treatment of arachnoid cyst of the fourth ventricle using a flexible endoscope via the anterior horn of the lateral ventricle. This method can be one of the treatments of the arachnoid cysts of the fourth ventricle.
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http://dx.doi.org/10.1016/j.wneu.2018.12.226DOI Listing
January 2019

Quantitative Blood Flow Assessment by Multiparameter Analysis of Indocyanine Green Video Angiography.

World Neurosurg 2018 Aug 3;116:e187-e193. Epub 2018 May 3.

Department of Neurosurgery, Asahikawa Medical University, Hokkaido, Japan. Electronic address:

Background: Measurements of quantitative blood flow are crucial during brain vascular surgery. Indocyanine green video angiography (ICG-VAG) is an accepted method of blood flow visualization; however, quantitative techniques have not yet been established. Thus, the aim of this study was to further develop ICG analysis for visualizing intraoperative flow changes.

Methods: We conducted basic experiments and clinical investigations to establish a relationship between ICG-VAG and measured blood flow. We evaluated several parameters and identified optimal indicators that precisely reflect blood (or fluid) flow. Both in vitro and in vivo studies were performed to calculate the interval between baseline and the intensity peak (Grad) and to measure actual flow rate.

Results: Grad and actual flow rate showed good exponential correlation, with R values of 0.90 in vitro and 0.82 in vivo. In a representative patient (case 3), we performed intraoperative flow analysis using FlowInsight, which identified a marked elevation in Grad on the brain surface. Because this observation is predictive of brain hyperperfusion, we used these data to carefully manage blood pressure postoperatively.

Conclusions: Grad is the optimum parameter for estimating flow conditions. Although ICG-VAG provides only visual profiles of blood circulation in the brain, this procedure has the potential to be widely used in clinical situations. ICG-based flow measurement can be used to identify normal and abnormal blood flow conditions, such as graft flow and vascular pathology. The novelty of this technique is that the fluorescence intensity of Grad enables surgeons to quantitatively measure real blood flow.
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http://dx.doi.org/10.1016/j.wneu.2018.04.148DOI Listing
August 2018
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