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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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Open-heart surgery in an infant with heterozygous factor VII deficiency

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INTERACTIVE CARDIOVASCULAR AND THORACIC SURGERY 2010年第6期10卷 1037-1038页

作者： Matsuhisa, Hironori Yoshimura, Naoki niimi, hideki Ichida, Fukiko Toyama Univ Dept Surg 1 Toyama 9300194 Japan Toyama Univ Dept Lab Med Toyama 9300194 Japan Toyama Univ Dept Pediat Toyama 9300194 Japan

A four-month-old male with Taussig-Bing anomaly and multiple ventricular septal defects underwent an open-heart palliative procedure. He suffered from massive postoperative gastrointestinal bleeding. Heterozygous Arg402Stop-related factor VII deficiency was detected by genomic examinations. When he was 14 months old, a subsequent open-heart surgery with replacement therapy of recombinant factor VIIa was performed without any bleeding or thromboembolic complications. Although heterozygous factor VII deficiency is generally recognized as clinically asymptomatic, this latent bleeding disorder can appear perioperatively or postoperatively in patients who undergo cardiopulmonary bypass procedures. Consequently, the prophylactic replacement therapy with recombinant factor VII is recommended during cardiac operations. (C) 2010 Published by European Association for Cardio-Thoracic Surgery. All rights reserved.

关键词： Pediatric Congenital heart disease Genes/polymorphisms Perioperative care

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Purulent lymphadenitis due to Halomonas hamiltonii: a case report

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INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES 2022年 125卷 145-148页

作者： Uejima, Yoji Oh-Ishi, Tsutomu Kitajima, Isao niimi, hideki Saitama Childrens Med Ctr Div Infect Dis & Immunol Saitama Japan Univ Toyama Grad Sch Med & Pharmaceut Sci Dept Clin Lab & Mol Pathol Toyama Japan Saitama Childrens Med Ctr Div Infect Dis & Immunol 1-2 ShintoshinChuou ku Saitama 3308777 Japan

Halomonas hamiltonii is a gram-negative rod bacterium isolated from highly saline environments. H. hamiltonii has rarely been reported as a human pathogen. Herein, we present the first case report of a purulent lymphadenitis caused by H. hamiltonii worldwide. The patient was a previously healthy girl aged 1 year who was referred to our hospital for left axillary lymphadenitis. Although oral amoxicillin was administered, lymphadenitis did not improve, and an abscess developed. After incision and drainage, the abscess was reduced. No recurrence of lymphadenitis was observed. The pus culture was negative. However, the 16S ribosomal DNA was amplified by the melting temperature mapping method. The amplified 16S ribosomal DNA sequence revealed 99.7% identity of H. hamiltonii . To the best of our knowledge, this is the first case of H. hamiltonii infection in a lymph node. This pathogen should be considered when diagnosing purulent lymphadenitis in healthy patients with lymphadenopathy of unknown origin. & COPY;2022 The Author(s). Published by Elsevier Ltd on behalf of International Society for Infectious Diseases. This is an open access article under the CC BY-NC-ND license ( http://***/licenses/by-nc-nd/4.0/ )

关键词： Lymphadenitis Halomonas Lymphadenopathy Case report

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Rapid detection of neonatal sepsis using the melting temperature mapping method

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PEDIATRICS AND NEONATOLOGY 2023年第5期64卷 617-618页

作者： Tamura, Kentaro niimi, hideki Nagaoka, Mitsuhide Yoshida, Taketoshi Toyama Univ Hosp Maternal & Perinatal Ctr Div Neonatol Toyama Japan Toyama Univ Hosp Clin Lab Ctr Toyama Japan Toyama Univ Hosp Maternal & Perinatal Ctr Div Neonatol 2630 Sugitani Toyama 9300194 Japan

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Time-dependent changes in cell population data obtained using Sysmex XN-series hematology analyzer in bacterial infections

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JOURNAL OF INFECTION AND CHEMOTHERAPY 2024年第10期30卷 983-988页

作者： Kaneda, Makito Nagaoka, Ro Yoshida, Rinako Iwasaki, Yosuke niimi, hideki Yamamoto, Yoshihiro Toyama Univ Hosp Dept Clin Infect Dis 2630 Sugitani Toyama 9300194 Japan Toyama Univ Hosp Dept Clin Lab & Mol Pathol Toyama Japan Sysmex Corp Sci Affairs Med & Sci Affairs Kobe Hyogo Japan

Background: Time-dependent changes in cell populations during acute bacterial infections remain unclear. We assessed time-dependent changes in fluorescent light intensity of the neutrophil area (NE-SFL) and fluorescent light distribution width index of the neutrophil area (NE-WY) and their association with sepsis and bacteremia. Methods: Patients with acute bacterial infections were enrolled in this prospective, observational cohort study. Blood samples were collected from all patients at the onset of bacterial infections (day 0) and on days 1 and 3. Microbiological evaluation included the examination of blood bacterial load using PCR. Cell population data were assessed using an automated hematology analyzer (Sysmex series XN-2000). Results: Forty-three participants with acute bacterial infections were enrolled in the study. Twenty-five participants developed definite sepsis. All the participants improved after the onset of infection. NE-WY levels showed significant time-dependent changes in participants with sepsis, peaking on day 0 and significantly decreasing until day 3, whereas these changes were not statistically significant for NE-SFL. A significant correlation with the Sequential Organ Failure Assessment score was observed with NE-WY and NE-SFL in the entire cohort on days 0 and 1. However, only NE-WY showed a significant correlation with blood bacterial load on days 0 and 1. Conclusion: This study demonstrated that NE-WY elevation in sepsis peaked earlier than NE-SFL, which may partly reflect the early bacterial invasion into circulation. These findings advocate caution in interpreting cell population data values as sepsis biomarkers and propose the potential of NE-WY as a therapeutic indicator.

关键词： XN-Series Bacteremia Cell population data Melting temperature mapping method Sepsis

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Gas6, a new regulator of chondrogenic differentiation from mesenchymal cells

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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 2007年第4期357卷 997-1003页

作者： Motomura, Hiraku niimi, hideki Sugimori, Kazuhito Ohtsuka, Toshihisa Kimura, Tomoatsu Kitajima, Isao Toyama Univ Fac Med Grad Sch Med & Pharmaceut Sci Dept Clin Lab & Mol Pathol Toyama 9300194 Japan Toyama Univ 21st Century COE Program Toyama 9300194 Japan Toyama Univ Fac Med Dept Orthopaed Surg Toyama 9300194 Japan

The mesenchymal cell line C3H10T1/2 can be preferentially induced toward chondrogenesis by culturing as a micromass in the presence of bone morphogenetic protein 2. To screen new regulator genes for chondrogenic differentiation, we performed differential display polymerase chain reaction and identified growth arrest-specific 6 (Gas6) as a gene that was clearly downregulated by this induction of chondrogenic differentiation. Blockage of Gas6 mRNA expression by siRNA remarkably enhanced the chondrogenic differentiation, while stimulation with recombinant Gas6 inhibited the mRNA expressions of type 11 collagen (Col2a1) and aggrecan. Gas6 signaling activated the phosphorylation of ERK1/2, SAPK/JNK, and Akt, but not p38 MAPK. These results suggest that Gas6 negatively regulates chondrogenic differentiation, at least through the MAPK pathway. (c) 2007 Elsevier Inc. All rights reserved.

关键词： Gas6 chondrogenesis differentiation C3H10T1/2 cell MAPK

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Melting temperature mapping method using imperfect-match linear long probes

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SCIENTIFIC REPORTS 2024年第1期14卷 1-11页

作者： Ootsuki, Shinya niimi, hideki Ueno, Tomohiro Mori, Masashi Tabata, Homare Minami, Hiroshi Kitajima, Isao Toyama Univ Hosp Cell Therapy Ctr Clin Lab & Transfus Med 2630 Sugitani Toyama 9300194 Japan Osaka Univ Hosp Lab Clin Invest Suita 5650871 Japan Ishikawa Prefectural Univ Res Inst Bioresources & Biotechnol Nonoichi Ishikawa 9218836 Japan Hokkaido Mitsui Chem Inc Life Sci Ctr Kushiro Hokkaido 0730138 Japan Univ Toyama Toyama 9308555 Japan

Identifying pathogenic microorganisms as early as possible is critical for selecting the appropriate antimicrobial therapy in infected patients. We previously reported the development of the Tm mapping method for identifying a broad range of pathogenic bacteria within 3 h of blood collection. However, the Tm mapping identification requires an analytical instrument with a tube-to-tube variation of no more than 0.1 degrees C, so we can only use a few instruments that have such high thermal accuracy. To address the problem, we developed the improved Tm mapping method using imperfect-match linear long quenching probes (IMLL Q-probes). Using IMLL Q-probes, almost all commercially available analytical instruments can be used for the Tm mapping method. Some bacterial species cannot be narrowed down to one species, but they can at least be narrowed down to the genus level. The Tm mapping method using IMLL Q-probes is useful for deciding on antimicrobial therapy in infected patients.

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Refractory tenosynovitis caused by Mycobacterium virginiense required repeated debridement: A case report and literature review

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Diagnostic Microbiology and Infectious Disease 2025年第1期113卷 116874-116874页

作者： Sekiguchi, Takafumi Kawasuji, Hitoshi Zukawa, Mineyuki Sugie, Nana Sugie, Kazushige Morita, Mika Nagaoka, Kentaro Morinaga, Yoshitomo niimi, hideki Yamamoto, Yoshihiro Department of Laboratory Medicine Toyama University Hospital 2630 Sugitani Toyama 930-0194 Japan Department of Clinical Infectious Diseases Toyama University Graduate School of Medicine and Pharmaceutical Sciences University of Toyama 2630 Sugitani Toyama 930-0194 Japan Department of Orthopaedic Surgery Faculty of Medicine University of Toyama 2630 Sugitani Toyama 930-0194 Japan Department of Microbiology Toyama University Graduate School of Medicine and Pharmaceutical Sciences University of Toyama 2630 Sugitani Toyama 930-0194 Japan

Mycobacterium virginiense is a slow-growing mycobacterium first described in 2016 as a novel species within the Mycobacterium terrae complex. To date, 8 cases of M. virginiense tenosynovitis or osteomyelitis have been reported, though none have been relapsed cases. A previously healthy 70-year-old Japanese man presented to a local orthopedic clinic with pain, swelling, and stiffness in his right middle finger. Initially diagnosed with trigger finger, he received multiple steroid injections. He was ultimately diagnosed with M. virginiense tenosynovitis. Despite synovectomy and antimicrobial therapy with ethambutol, rifampicin, and clarithromycin, a relapse occurred, necessitating a second tenosynovectomy, which led to remission. Antimicrobial therapy continued for approximately 20 months without recurrence. This case illustrates that M. virginiense can cause refractory tenosynovitis even in immunocompetent hosts. Delayed diagnosis and repeated local steroid injections may have contributed to disease progression and treatment failure in M. virginiense tenosynovitis. © 2025 The Author(s)

关键词： Hand Mycobacterium virginiense Non-tuberculous mycobacteria Tenosynovitis

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Disseminated nocardiosis complicated by multiple abscesses of the brain and lower limbs diagnosed by the melting temperature mapping method: A case report

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JOURNAL OF ORTHOPAEDIC SCIENCE 2023年第6期28卷 1497-1500页

作者： Ikeda, Shinsuke Uchiyama, Katsufumi Moriya, Mitsutoshi Sakurai, Keizo Nihonyanagi, Shin niimi, hideki Takaso, Masashi Kitasato Univ Dept Orthopaed Surg Sch Med 1-15-1 KitasatoMinami Ku Sagamihara Kanagawa 2520374 Japan Kurokouchi Hosp 17-36 YutakachoMinami Ku Sagamihara Kanagawa 2520305 Japan Kitasato Univ Hosp Dept Clin Lab 1-15-1 KitasatoMinami Ku Sagamihara Kanagawa 2520375 Japan Kitasato Univ Hosp Dept Infect Control & Prevent 1-15-1 KitasatoMinami Ku Sagamihara Kanagawa 2520375 Japan Univ Toyama Grad Sch Med & Pharmaceut Sci Res Dept Clin Lab & Mol Pathol 2630 Sugitani Toyama 9300194 Japan

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A 2x2 paralleling GaN half-bridge power module for high-density 500kHz 3.3kW CLLC converter 38

A 2x2 paralleling GaN half-bridge power module for high-dens...

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IEEE Applied Power Electronics Conference and Exposition (APEC)

作者： Otsu, Satoshi Ujita, Shinji Tanaka, Kenichiro Maede, Masato niimi, hideki Ochi, Shozo Panason Ind Co Ltd Engn Div Moriguchi Osaka 5708501 Japan Panason Ind Co Ltd Mfg Innovat Ctr Kadoma Osaka 5718506 Japan

ISBN: (纸本)9781665475396

A 2x2 paralleling GaN half-bridge power module is developed, where four GaN dies are encapsulated with minimizing parasitic inductances. It is suitable for high-power, highly efficient and high-density power converters under high frequency operation. It is confirmed that the module board has small parasitic components by electromagnetic field simulation. In this paper, we demonstrate a 3.3 kW CLLC converter which is operated at the switching frequency of around 500 kHz employing the developed 2x2 paralleling GaN half-bridge power module. The power density and the peak efficiency of the CLLC board are 8.8 W/ cm(3) and 97.1 % respectively, which correspond to 25 % and 0.3 % improvement compared with the case in which four discrete GaN power transistors are employed, respectively. The obtained improvements are attributable to the smaller parasitic inductances of the developed GaN module.

关键词： GaN Module DC-DC Converter LLC CLLC Half-bridge Power Density High Frequency High Power Parasitic inductance PCB Transformer Zero-voltage switching Zero-current switching Soft-switching

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