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High-throughput sperm screening using one-step RT-qPCR: Improvement and re-evaluation

ANALYTICAL BIOCHEMISTRY 2025年 698卷 115727页

作者： kubo, seiji Amai, Keito Nakano, Fumitaka Tanaka, Jin Niimi, Hideki Univ Toyama Fac Med Dept Clin Lab & Mol Pathol Acad Assembly 2630 Sugitani Toyama 9300194 Japan Ishikawa Prefectural Police Headquarters Forens Sci Lab 1-1 Kuratsuki Kanazawa 9208553 Japan

Sperm identification is crucial in sexual assault cases. While microscopic analysis is the gold standard for sperm detection, it is a laborious procedure even for trained personnel. Reverse transcription-quantitative real-time PCR (RT-qPCR) can enhance the screening by detecting sperm-specific mRNA markers, such as protamine 2 (PRM2). This study aimed to develop a one-step RT-qPCR assay targeting PRM2 mRNA. Our assay was capable of detecting as low as 0.01 mu L of semen with high specificity and demonstrated successful detection of PRM2 mRNA in simulated-case samples. Owing to the simple workflow involved, our assay requires <30 min for RNA extraction and <60 min for RT-qPCR. Our assay enables high-throughput sperm screening and offers a promising strategy for enhancing the workflow of sexual assault cases.

关键词： Semen Sperm PRM2 RT-qPCR

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Valgus positioning of the femoral component does not compromise short-term clinical outcomes in fixed-bearing unicompartmental knee arthroplasty

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EUROPEAN JOURNAL OF ORTHOPAEDIC SURGERY AND TRAUMATOLOGY 2025年第1期35卷 1-6页

作者： Anjiki, Kensuke Nakano, Naoki kubo, seiji Ishida, Kazunari Ikuta, Kemmei Tsubosaka, Masanori Kamenaga, Tomoyuki Kuroda, Yuichi Hayashi, Shinya Kuroda, Ryosuke Matsumoto, Tomoyuki Kobe Univ Grad Sch Med Dept Orthoped Surg Kobe Japan Anshin Hosp Dept Orthopaed Surg Kobe Japan Kubo Orthoped Clin Akashi Japan Kobe Kaisei Hosp Dept Orthopaed Surg Kobe Japan

Purpose: In unicompartmental knee arthroplasty (UKA), the tibial component tends to be placed in a mildly varus position because of the expected improvement in stress distribution and load transfer across the tibia. With the spacer block technique in UKA, if a tibial osteotomy is performed in the varus position, the femoral component is placed in the valgus position. Therefore, this study aimed to evaluate the impact of valgus positioning of the femoral component on the short-term postoperative clinical outcomes in fixed-bearing UKA using the spacer block technique. Methods: We analyzed 77 knees of 70 patients who underwent fixed-bearing UKA using the spacer block technique for medial compartment osteoarthritis and spontaneous osteonecrosis of the knee. Patients were categorized based on femoral component alignment: the neutral group (-3 degrees to + 3 degrees, mean 0.5 degrees +/- 1.7 degrees valgus) and the valgus group (> 3 degrees, mean 5.7 degrees +/- 1.7 degrees valgus). Postoperative outcomes, including the femorotibial angle (FTA), component sagittal alignment, knee range of motion, and 2011 Knee Society Score (KSS), were compared between the two groups. Results: No significant differences were found between the neutral and valgus groups regarding the postoperative FTA, component sagittal alignment, knee range of motion (neutral vs. valgus group;extension,degrees: - 1.9 +/- 2.9 vs. - 1.6 +/- 2.9;flexion,degrees: 133.0 +/- 6.9 vs. 131.3 +/- 7.9), or 2011 KSS (objective knee indicators: 95.0 +/- 8.9 vs. 96.8 +/- 3.5;functional activities: 78.9 +/- 15.3 vs. 78.2 +/- 12.9). Conclusion: These findings support the safety of valgus positioning in fixed-bearing UKA, offering flexibility in component placement without compromising short-term clinical outcomes.

关键词： Unicompartmental knee arthroplasty Valgus positioning of the femoral component Spacer block technique Clinical outcome

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Topology Optimization For The Duct Flow Problems In Laminar And Turbulent Flow Regimes

Topology Optimization For The Duct Flow Problems In Laminar ...

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作者： seiji kubo Kyoto University

学位级别：博士

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Rapid screening method for male DNA by using the loop-mediated isothermal amplification assay

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INTERNATIONAL JOURNAL OF LEGAL MEDICINE 2018年第4期132卷 975-981页

作者： Kitamura, Masashi kubo, seiji Tanaka, Jin Adachi, Tatsushi Ishikawa Prefectural Police HQ Forens Sci Lab 1-1 Kuratsuki Kanazawa Ishikawa 9208553 Japan Kanazawa Univ Grad Sch Med Sci Div Pharmaceut Sci Lab Mol Pharmacognosy Kakuma Machi Kanazawa Ishikawa 9201192 Japan Kaneka Corp 1-8 Miyamae ChoTakasago Cho Takasago Hyogo 6768688 Japan

Screening for male-derived biological material from collected samples plays an important role in criminal investigations, especially those involving sexual assaults. We have developed a loop-mediated isothermal amplification (LAMP) assay targeting multi-repeat sequences of the Y chromosome for detecting male DNA. Successful amplification occurred with 0.5 ng of male DNA under isothermal conditions of 61 to 67 A degrees C, but no amplification occurred with up to 10 ng of female DNA. Under the optimized conditions, the LAMP reaction initiated amplification within 10 min and amplified for 20 min. The LAMP reaction was sensitive at levels as low as 1-pg male DNA, and a quantitative LAMP assay could be developed because of the strong correlation between the reaction time and the amount of template DNA in the range of 10 pg to 10 ng. Furthermore, to apply the LAMP assay to on-site screening for male-derived samples, we evaluated a protocol using a simple DNA extraction method and a colorimetric intercalating dye that allows detection of the LAMP reaction by evaluating the change in color of the solution. Using this protocol, samples of male-derived blood and saliva stains were processed in approximately 30 min from DNA extraction to detection. Because our protocol does not require much hands-on time or special equipment, this LAMP assay promises to become a rapid and simple screening method for male-derived samples in forensic investigations.

关键词： Loop-mediated isothermal amplification Forensic science Sex identification On-site detection

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A topology optimization method for a coupled thermal-fluid problem using level set boundary expressions

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INTERNATIONAL JOURNAL OF HEAT AND MASS TRANSFER 2015年 81卷 878-888页

作者： Yaji, Kentaro Yamada, Takayuki kubo, seiji Izui, Kazuhiro Nishiwaki, Shinji Kyoto Univ Dept Mech Engn & Sci Nishikyo Ku Kyoto 6158540 Japan IHI Corp R&D Technol Ctr Numer Engn Dept Isogo Ku Yokohama Kanagawa 2358501 Japan

This paper presents a topology optimization method for a coupled thermal-fluid problem based on the two- and three-dimensional steady-state Navier-Stokes and energy equations. In this research, the optimization problem is formulated as a heat exchange maximization problem to obtain structures that function as high-performance cooling devices. Such devices, for example, liquid-cooled heat sinks, have recently attracted considerable attention as an engineering application for thermal cooling devices. The proposed optimization method employs level set boundary expressions and a Tikhonov-based regularization scheme enables qualitative control of the geometric complexity of the optimal configurations. Using the developed methodology, we provide two- and three-dimensional numerical examples that confirm the applicability, from an engineering standpoint, of the optimization method for the design of cooling devices. (C) 2014 Elsevier Ltd. All rights reserved.

关键词： Topology optimization Level set method Phase field method Thermal-fluid Heat sink

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A level set-based topology optimization method for optimal manifold designs with flow uniformity in plate-type microchannel reactors

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STRUCTURAL AND MULTIDISCIPLINARY OPTIMIZATION 2017年第4期55卷 1311-1327页

作者： kubo, seiji Yaji, Kentaro Yamada, Takayuki Izui, Kazuhiro Nishiwaki, Shinji IHI Corp Yokohama Kanagawa 2358501 Japan Kyoto Univ Dept Mech Engn & Sci Nishikyo Ku Kyoto 6158540 Japan

This paper proposes an optimal design method for plate-type microchannel reactor manifolds, using a level set-based topology optimization method that targets flow uniformity among the microchannels. For plate-type microchannel reactors, manifold designs must consider both the uniformity of flow among the microchannels and minimization of pressure drop in the device. To address these design requirements, the pressure drop in the device is defined as the objective functional to be minimized under a flow rate inequality constraint, defined as the deviation in flow rate among the microchannels during the optimization. In addition, we propose a comparably simple and stable augmented Lagrangian method using an exponential function, to determine the Lagrange multiplier that acts to satisfy the flow rate inequality constraint. To demonstrate the utility of our proposed method, two-dimensional Z-type and U-type manifold design problems are presented in the numerical examples.

关键词： Topology optimization Level set method Flow uniformity Flow rate inequality constraint Manifold shape

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Intra-operative joint gap kinematics in unicompartmental knee arthroplasty

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CLINICAL BIOMECHANICS 2013年第1期28卷 29-33页

作者： Matsumoto, Tomoyuki Muratsu, Hirotsugu kubo, seiji Kuroda, Ryosuke Kurosaka, Masahiro Kobe Univ Grad Sch Med Dept Orthopaed Surg Kobe Hyogo 6500017 Japan Steel Mem Hirohata Hosp Dept Orthopaed Surg Himeji Hyogo Japan

Background: The use of an offset type tensor for total knee arthroplasty that can be set with patellofemoral joint reduction and femoral component placement enables surgeons to assess soft tissues in the physiological postoperative knee condition, showing different kinematic pattern of soft tissues in varus osteoarthritic knees between cruciate-retaining and posterior-stabilized total knee arthroplasty. However, gap kinematics in unicompartmental knee arthroplasty is unclear. Methods: Using a newly developed tensor that is designed to assess soft tissue balance throughout the full range of motion with femoral component placement, we assessed the intra-operative joint gap measurements of unicompartmental knee arthroplasties performed at 0, 10, 30, 45, 60, 90, 120 and 135 degrees of flexion in 20 osteoarthritic patients. In addition, the kinematic pattern of unicompartmental knee arthroplasty was compared with those of cruciate-retaining and posterior-stabilized total knee arthroplasty that were calculated as medial compartment gap from the previous series of this study. Findings: While the joint gap measurements of unicompartmental knee arthroplasties increased from full extension to extension (10 degrees of flexion), these values remained constant throughout the full range of motion. Of note, the gap values of cruciate-retaining total knee arthroplasty were significantly smaller from midrange to deep flexion compared with posterior-stabilized total knee arthroplasty, and furthermore unicompartmental knee arthroplasty showed a significantly smaller gap from extension to midrange flexion compared with cruciate-retaining total knee arthroplasty. Interpretation: Accordingly, we conclude that the intra-operative joint gap kinematic pattern in unicompartmental knee arthroplasty differs from the pattern in total knee arthroplasty. (C) 2012 Elsevier Ltd. All rights reserved.

关键词： Unicompartmental knee arthroplasty Soft tissue balance Total knee arthroplasty Cruciate-retaining Posterior stabilized

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Improving the etching performance of high-aspect-ratio contacts by wafer temperature control Uniform temperature design and etching rate enhancement

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PRECISION ENGINEERING-JOURNAL OF THE INTERNATIONAL SOCIETIES FOR PRECISION ENGINEERING AND NANOTECHNOLOGY 2016年 44卷 87-92页

作者： Tandou, Takumi kubo, seiji Yokogawa, Ken'etsu Negishi, Nobuyuki Izawa, Masaru Hitachi High Technol Corp Elect Device Syst Business Grp 794 Higashitoyoi Kudamatsu City Yamaguchi 7440002 Japan Hitachi Reftechno Inc Refrigerat & Elect Syst Design Dept 709-2 Tomita Tochigi City Tochigi 3294404 Japan

A novel wafer temperature control system using direct expansion cycles is developed to improve etching performance. This system enables rapid temperature control of a wafer with low power consumption. In a previous report, we confirmed that the etching rate and mask selectivity of high-aspect-ratio contact etching could be increased by around 6% and 14%, respectively, by controlling the temperature of the wafer during the etching process. In this study, an advanced wafer temperature control system that realizes not only rapid response but also uniform wafer cooling is developed, and a new etching process that controls 02 gas flow rate as well as wafer temperature during etching is evaluated to decrease the etching rate depression of high-aspect-ratio contact etching. As a result, a rate of wafer temperature change of 1 degrees C/s and uniformity of +/- 0.7% with a coefficient of performance exceeding 3 is achieved over a wafer with a diameter of 300 mm during the etching process. Furthermore, etching rate depression in C4F6/Ar/O-2 plasma is decreased from 14.4% to 7.8% for a sample with a diameter of 100 nm and aspect ratio of 30. (C) 2015 Elsevier Inc. All rights reserved.

关键词： Plasma etching Direct expansion system Temperature control Etching rate Coefficient of performance

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AERODYNAMIC PERFORMANCE OF NOVEL LIGHTWEIGHT TURBINE BLADE

AERODYNAMIC PERFORMANCE OF NOVEL LIGHTWEIGHT TURBINE BLADE

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ASME Turbo Expo: Turbine Technical Conference and Exposition

作者： Okita, Yoji Nita, Kozo kubo, seiji IHI Corp Corp Res & Dev Yokohama Kanagawa Japan IHI Corp Aeroengine & Space Operat Tokyo Japan

ISBN: (纸本)9780791849705

The primary contribution of this research is to clarify the aerodynamic performance of a novel lightweight turbine blade with internal cooling passage and external film cooling, which is invented aiming at drastic weight reduction of a cooled blade. With a considerably thinner airfoil, a significant separation region is formed along the pressure side, and therefore aerodynamic performance with such a flow field should be investigated. First, the lightweight cooled airfoil is designed. In the design process, a conventional thick airfoil is first defined as a baseline. With the baseline airfoil, only the mid and rear part of pressure side profile is redesigned to thin the airfoil without any change in the suction side geometry. The airfoil geometry is optimized so as not to bring significant aerodynamic loss increase. In this numerical optimization, the airfoil shape is gradually changed and evaluated step by step. In every step, an adjoint variable method is used to seek better airfoil shape, and then the generated new shape is evaluated with full RANS calculation. This iteration is repeated until any further recognizable weight reduction cannot be obtained without sensitive pressure loss increase and/or the airfoil shape reaches some geometrical constraints. The resultant optimized airfoil is approximately 20% lighter than the baseline hollow airfoil without any noticeable change in aerodynamic loss in the numerical solution. Next, the optimized airfoil is tested in a high speed linear cascade rig to verify its aerodynamic performance. The baseline airfoil is also tested for comparison. The rig is composed of six airfoil passages. The compressed air is supplied to the cascade and discharges to the atmospheric exhaust chamber. The air is also heated up to about 540 K upstream of the cascade. The cascade exit Mach number at the design point is 1.25, while in the experiment other several off-design conditions are also tested to check if there is any Mach number sensiti

关键词： Airfoils

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Bayesian Uncertainty Reduction of Generalised k-ω Turbulence Model for Prediction of Film-Cooling Effectiveness

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INTERNATIONAL JOURNAL OF COMPUTATIONAL FLUID DYNAMICS 2022年第2期36卷 152-166页

作者： Misaka, Takashi Nakazawa, Takashi Obayashi, Shigeru kubo, seiji Asaumi, Norio Ideta, Takeomi Natl Inst Adv Ind Sci & Technol Koto Ku Tokyo Japan Osaka Univ Toyonaka Osaka Japan Tohoku Univ Sendai Miyagi Japan IHI Corp Koto Ku Tokyo Japan IHI Scube Co Ltd Koto Ku Tokyo Japan

In this study, Reynolds-averaged Navier-Stokes (RANS) simulations were carried out to predict the film cooling effectiveness of an inclined round jet in crossflow using turbulence model parameters optimised based on measurement data. The posterior distributions of the generalised k-omega (GEKO) turbulence model parameters were estimated using a computationally efficient surrogate model with the Markov chain Monte Carlo method, which provides a framework for probabilistic parameter estimation based on measurement data. The results show that using the maximum a posterior parameters for a blowing ratio of 0.5 gives better predictions than using the default parameters of the GEKO model. The estimated parameters were then applied to flows with a higher blowing ratio and different hole geometry to evaluate the generalisation performance. In both cases, the results were improved by properly predicting the spread of the cooling flow.

关键词： Gas turbine film cooling RANS turbulence model Bayesian calibration data assimilation

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