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Development and evaluation of a SYBR Green-based RT-qPCR assay with a specific primer set for tomato seed testing against tomato brown rugose fruit virus

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JOURNAL OF GENERAL PLANT PATHOLOGY 2025年第3期91卷 160-170页

作者： ota, emi Shinosaka, Hibiki Ishibashi, Kazuhiro Takeyama, Sawana Matsuyama, Momoko Tomitaka, Yasuhiro Matsushita, Yosuke Osaki, Kohei Kubota, Kenji NARO Inst Plant Protect Div Core Technol Pest Control Res 2-1-18 Kannondai Tsukuba Ibaraki 3058666 Japan NARO Ctr Seeds & Seedlings Dept DUS Test & Seed Inspect 2-2 Fujimoto Tsukuba Ibaraki 3050852 Japan NARO Inst Agrobiol Sci Div Plant Mol Regulat Res 2-1-2 Kannondai Tsukuba Ibaraki 3058602 Japan NARO Inst Plant Protect Div Crop Pest Control Res 2-1-18 Kannondai Tsukuba Ibaraki 3058666 Japan

Since its first occurrence in Israel and Jordan in 2014 and 2015, the tomato brown rugose fruit virus (ToBRFV) has become one of the most concerning pathogens affecting tomatoes and other crops worldwide. Its rapid spread is believed to result from the international trade of contaminated seeds and its seed transmissibility, underscoring the critical importance of seed health testing for ToBRFV to prevent further dissemination of the virus. To this end, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) protocols employing TaqMan probe chemistry have been widely adopted. However, the development of RT-qPCR protocols for ToBRFV seed testing using SYBR Green chemistry remains limited. The SYBR Green method offers the advantage of distinguishing ToBRFV from other tobamoviruses through melt curve analysis. In this study, we developed a SYBR Green-based RT-qPCR detection method using newly designed primer sets, which demonstrated high specificity for ToBRFV and sufficient sensitivity. While this protocol requires further optimization and validation for application in routine seed testing, it establishes a foundational approach for SYBR Green-based RT-qPCR seed testing. Additionally, this study raises an important question regarding the relationship between RT-qPCR results in seed tests and the likelihood of virus contamination or transmission via seeds.

关键词： ToBRFV Tobamoviruses SYBR green RT-qPCR Melt curve Tomato

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A multicenter randomized controlled trial evaluating the effect of the use of an anti-adhesion barrier for diverting ileostomy on the multidimensional workload in minimally invasive surgery for rectal cancer (YCOG 2005: The ADOBARRIER study)

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ANNALS OF GASTROENTEROLOGICAL SURGERY 2025年第0期

作者： ota, emi Watanabe, Jun Suwa, Yusuke Numata, Masakatsu Suwa, Hirokazu Ohya, Hiroki Nakagawa, Kazuya Ozawa, Mayumi Endo, Itaru Yokosuka Kyosai Hosp Dept Surg Yokosuka Japan Kansai Med Univ Dept Colorectal Surg 2-5-1 Shin machi Hirakata Osaka 5731010 Japan Yokohama City Univ Gastroenterol Ctr Med Ctr Dept Surg Yokohama Japan Yokohama City Univ Grad Sch Med Dept Gastroenterol Surg Yokohama Japan

Aim: The purpose of this study was to assess whether the use of spray-type anti-adhesion material during diverting ileostomy construction could reduce the surgeon's multidimensional workload, the degree of adhesion, and the operation time in ileostomy closure. Methods: Patients diagnosed with rectal cancer, who were scheduled for laparoscopic or robotic rectal surgery followed by diverting ileostomy, were single-blindly (patient-blind), randomly assigned to either the AdSpray (TM) arm or the control arm. The primary endpoint was the multidimensional workload of the ileostomy closure operator (SURG-TLX value). Results: Between January 2020 and December 2022, 126 patients were enrolled. Five patients were excluded and a total of 121 patients (control arm, n = 60;AdSpray (TM) arm, n = 61) were analyzed. The baseline factors were well balanced between the two arms. Regarding SURG-TLX in ileostomy closure, operators in the AdSpray (TM) arm required a significantly lower overall workload than those in the control arm (AdSpray (TM) arm, 28.1;control arm, 58.9;p < 0.001). Mental, physical, and temporal demands, task complexity, situation stress, and distractions were significantly lower in the AdSpray (TM) arm (p < 0.001). Operative time was significantly shorter in the AdSpray (TM) arm (AdSpray (TM) arm, 58 min;control arm, 65 min;p = 0.040). The degree of adhesion (p < 0.001) and extent of intra-abdominal adhesions (p < 0.001) in ileostomy closure were significantly lower in the AdSpray (TM) arm. Conclusions: The use of spray-type anti-adhesion material was associated with a significantly lower SURG-TLX value, lower incidence of adhesion, less severe adhesion, and shorter operative time.

关键词： anti-adhesion barrier diverting ileostomy ileostomy closure rectal cancer SURG-TLX

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Secondary debulking surgery for para-aortic nodal recurrence in endometrial cancer requiring circumferential resection of the inferior vena cava

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GYNECOLOGIC ONCOLOGY 2018年第1期149卷 221-222页

作者： Kato, Kazuyoshi Okamoto, Shuhei ota, emi Takeshima, Nobuhiro Canc Inst Hosp Dept Gynecol Koutou Ku 3-8-31 Ariake Tokyo 1358550 Japan

Objective Recurrent endometrial cancer presenting as isolated lymph node metastases is considered suitable for secondary debulking surgery, since a complete cytoreduction can be performed in most cases. However, the resection of enlarged lymph nodes can result in severe intraoperative complications, including the laceration of major retroperitoneal vessels and increased blood loss [1]. Here, we report the details of a surgical technique for the resection of metastatic para-aortic lymph nodes with involvement of the inferior vena cava (NC) requiring circumferential resection of the IVC. Methods The subject was a 43-year-old woman who had been initially treated for endometrial cancer at another hospital. A lymphadenectomy was not performed during the primary surgery. The histopathologic results revealed G2 endometrioid adenocarcinoma. Following the surgery, she was treated with adjuvant chemotherapy, comprised of 6 cycles of paclitaxel and carboplatin. One year after the initial management, paraaortic lymph node recurrence was suspected. She was referred to our hospital at that time. A computed tomography (CT) examination revealed para-aortic adenopathy in the lateroaortic and retrocaval regions. No other lesions were observed. The patient subsequently underwent a secondary debulking surgery for diagnostic and therapeutic purposes. The lymph nodes around the inferior vena cava at the portion of the bifurcation to the iliac veins adhered to the vena cava. During the resection of these lymph nodes, the NC was injured and severe bleeding occurred. To achieve hemostatic control, proximal and distal clamps were applied to the injured portion of the IVC. During clamping of the IVC, changes in the vital signs, including the systolic blood pressure and the urinary volume, were not noted. As the metastatic nodes involved the posterior surface of the IVC wall, an en bloc resection of the lymph nodes with the involved portion of the IVC was performed at a level immediately above

关键词： Recurrent endometrial cancer Para-aortic lymphadenectomy Inferior vena cava Circumferential resection

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A next-generation-sequencing approach to diagnose viral diseases of greenhouse ranunculus

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JOURNAL OF GENERAL PLANT PATHOLOGY 2022年第2期88卷 134-137页

作者： Hayahi, Saki ota, emi Kurogi, Shuichi Matsushita, Yosuke Kanno, Yoshiaki Takeshita, Minoru Univ Miyazaki Miyazaki 8892192 Japan Miyazaki Agr Res Inst Miyazaki 8800212 Japan NARO Inst Plant Protect Ibaraki 3058666 Japan Minami Kyushu Univ Miyazaki 8850035 Japan

Viruses that infect Ranunculus asiaticus L., a globally important greenhouse ornamental, have consistently caused serious damage in Japan. We coupled nextgeneration-sequencing (NGS) with visual assessment of symptoms on leaf tissues, and found almost all of the diseases were caused by ranunculus mild mosaic virus (RanMMV) and ranunculus leaf distortion virus (RanLDV). These results were confirmed by reverse transcription-polymerase chain reaction specific to viral genes of ranunculus-infecting viruses. NGS with symptom assessment should be able to provide a rapid and practical diagnostic method focusing on RanMMV and RanLDV in Japan.

关键词： Ranunculus Next-generation-sequencing RanMMV RanLDV

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Incidence and risk factors of bowel dysfunction after minimally invasive rectal cancer surgery and discrepancies between the Wexner score and the low anterior resection syndrome (LARS) score

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SURGERY TODAY 2024年第7期54卷 763-770页

作者： ota, emi Nagasaki, Toshiya Akiyoshi, Takashi Mukai, Toshiki Hiyoshi, Yukiharu Yamaguchi, Tomohiro Fukunaga, Yosuke Japanese Fdn Canc Res Canc Inst Hosp Gastroenterol Ctr Dept Gastroenterol Surg 3-8-31 AriakeKoto Ku Tokyo 1358550 Japan

PurposeBowel dysfunction after sphincter-preserving-surgery (SPS) impacts quality of life. The Wexner score (WS) and the low anterior resection syndrome (LARS) score (LS) are instruments for assessing postoperative bowel dysfunction. We analyzed the incidence of and risk factors for each symptom and examined the discrepancies between the two *** total of 142 patients with rectal cancer, who underwent minimally invasive SPS between May, 2018 and July, 2019, were included. A questionnaire survey using the two scores was given to the patients 2 years after *** location and preoperative radiotherapy were independent risk factors for major LARS. Intersphincteric resection with a hand-sewn anastomosis (HSA) was an independent risk factor for high WS. Among the patients who underwent HSA, 82% experienced incontinence for liquid stools, needed to wear pads, and suffered lifestyle alterations. Of the 35 patients with minor LARS, only 1 had a high WS, and 80.0% reported no lifestyle alterations. Among the 75 patients with major LARS, 58.7% had a low WS and 21.3% reported no lifestyle *** results of this study provide practical data to help patients understand potential bowel dysfunction after SPS. The discrepancies between the WS and LS were clarified, and further efforts are required to utilize these scores in clinical practice.

关键词： Wexner score Low anterior resection syndrome score Sphincter-preserving-surgery Minimally invasive surgery Rectal cancer

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Cytoreductive surgery without intra-peritoneal chemotherapy for metachronous colorectal peritoneal metastases

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WORLD JOURNAL OF SURGICAL ONCOLOGY 2024年第1期22卷 205页

作者： ota, emi Fukunaga, Yosuke Mukai, Toshiki Hiyoshi, Yukiharu Yamaguchi, Tomohiro Nagasaki, Toshiya Akiyoshi, Takashi Japanese Fdn Canc Res Canc Inst Hosp Dept Gastroenterol Surg Tokyo Japan

Background Cytoreductive surgery and chemotherapy reportedly improve the prognosis of patients with metachronous peritoneal metastases. However, the types of peritoneal metastases indicated for cytoreductive surgery remains unclear. Therefore, we aimed to clarify the category of cases for which cytoreductive surgery would be effective and report the prognosis associated with cytoreductive surgery for metachronous peritoneal metastases. Methods This study included 52 consecutive patients who underwent cytoreductive surgery for metachronous peritoneal metastases caused by colorectal cancer between January 2005 and December 2018 and fulfilled the selection criteria. The median follow-up period was 54.9 months. Relapse-free survival was calculated as the time from cytoreductive surgery of metachronous peritoneal metastases to recurrence. Overall survival was defined as the time from cytoreductive surgery of metachronous peritoneal metastases to death or the end of the follow-up period. Results The 5-year relapse-free survival rate was 30.0% and the 5-year overall survival rate was 72.3%. None of the patients underwent hyperthermic intraperitoneal chemotherapy. The analysis indicated no potential risk factors for 5-year relapse-free survival. However, for 5-year overall survival, the multivariate analysis revealed that time to diagnosis of metachronous peritoneal metastases of < 2 years after primary surgery (hazard ratio = 4.1, 95% confidence interval = 2.0-8.6, p = 0.0002) and number of metachronous peritoneal metastases >= 3 (hazard ratio = 9.8, 95% confidence interval = 2.3-42.3, p = 0.002) as independent factors associated with a poor prognosis. Conclusions Long intervals of more than 2 years after primary surgery and 2 or less metachronous peritoneal metastases were good selection criteria for cytoreductive surgery for metachronous peritoneal metastases from colorectal cancer.

关键词： Metachronous peritoneal metastases Cytoreductive surgery 5-year relapse-free survival 5-year overall survival Colorectal cancer

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Effectiveness of nurse-led transitional care interventions for adult patients discharged from acute care hospitals: a systematic review and meta-analysis

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BMC NURSING 2025年第1期24卷 1-21页

作者： Sakashita, Chizuko Endo, emi ota, Erika Oku, Hiromi Kitasato Univ Hosp Nursing Dept 1-15-1 KitasatoMinami Ku Sagamihara Kanagawa Japan St Lukes Int Univ Grad Sch Nursing Doctoral Program Tokyo Japan Yokohama City Univ Med Ctr Nursing Dept Yokohama Japan St Lukes Int Univ Grad Sch Nursing Tokyo Japan

BackgroundWith the guidance of healthcare policy and advances in medical technology, the average length of stay in hospitals continues to decrease. In this context, expectations for nurse-led interventions for patients discharged home are increasing. However, few systematic reviews of nurse-led transitional care have focused on patients discharged from acute care hospitals. This systematic review aimed to assess the effects of nurse-led transitional care interventions on readmission rates, unscheduled outpatient-visit rates, and quality of life (QOL) of adult patients discharged from acute care hospitals, compared with usual *** electronic databases were searched for articles published through October 2023. Individual and cluster randomized controlled trials (RCTs) examining the effectiveness of nurse-led transitional care interventions were included. Independent reviewers performed study selection, data extraction, risk of bias assessment, and certainty of evidence using the GRADE *** RCTs were included. In a meta-analysis of RCTs with readmission rates as the outcome, readmission rates were significantly reduced in the intervention group when the data collection period exceeded 12 weeks (RR 0.67;95% CI, 0.49-0.92;P = 0.01;I-2 = 66%;certainty: moderate). The rate of emergency room visits was also significantly reduced in the intervention group (RR 0.63;95% CI, 0.49-0.81;P = 0.0003;I-2 = 0%;certainty: high). QOL measured with the SF-36 was significantly higher after 5 weeks (MD 1.27;95% CI, 0.52-2.02;P = 0.0009, I-2 = 0%;certainty: low) and after 6 weeks (MD 2.46;95% CI, 1.67-3.25;P = 0.00001;I-2 = 19%;certainty: low), both showing a possibility of improvement in the intervention group. However, the number of studies and samples included in the meta-analysis, particularly for readmission rates and QOL, were small, and the results should be interpreted with caution due to differences in subjects, institutions, and types of interventio

关键词： Meta-analysis Nurse-led intervention Quality of life Transitional care Readmission

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Laparoscopic extraperitoneal colostomy has a lower risk of parastomal hernia and bowel obstruction than transperitoneal colostomy

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INTERNATIONAL JOURNAL OF COLORECTAL DISEASE 2022年第6期37卷 1429-1437页

作者： ota, emi Yamaguchi, Tomohiro Nagasaki, Toshiya Fukuoka, Hironori Mukai, Toshiki Hiyoshi, Yukiharu Konishi, Tsuyoshi Akiyoshi, Takashi Fukunaga, Yosuke Japanese Fdn Canc Res Canc Inst Hosp Dept Gastroenterol Surg Tokyo Japan Yokohama City Univ Dept Gastroenterol Yokohama Kanagawa Japan Univ Texas MD Anderson Canc Ctr Dept Colon & Rectal Surg Houston TX USA

Purpose Several studies indicate that an extraperitoneal colostomy can prevent the development of a parastomal hernia (PSH) as compared to a transperitoneal colostomy. However, the clinical value of laparoscopic extraperitoneal colostomy, and its influence on bowel obstruction and PSH remain unclear. The present study aimed to clarify the impact of laparoscopic extraperitoneal colostomy on the development of a PSH and bowel obstruction. Methods This study included 327 consecutive patients who underwent laparoscopic abdominoperineal resection or Hartmann's procedure between January 2013 and December 2019 after fulfilling selection criteria. The incidence of a PSH (Clavien-Dindo classification >= grade I) and bowel obstruction (>= grade IIIa) in the transperitoneal and extraperitoneal route groups were analyzed using univariate and multivariate analysis. Results The patients were classified into transperitoneal (n = 222) and extraperitoneal (n = 105) route groups. The patient characteristics, except for body mass index and operative time, were comparable between the groups. A PSH and bowel obstruction occurred more frequently in the transperitoneal than in the extraperitoneal route group (17.1% vs. 1.9% and 15.3% vs. 6.7%, respectively;p < 0.01 and p = 0.03, respectively). The multivariate analysis showed that age >= 70 years, body mass index >= 22.4 kg/m(2), and a transperitoneal route were independent risk factors for the development of a PSH, and a transperitoneal route was an independent risk factor for bowel obstruction. Conclusions The transperitoneal route was identified as a risk factor for the development of both a PSH and bowel obstruction after laparoscopic abdominoperineal resection or Hartmann's procedure.

关键词： Bowel obstruction Extraperitoneal route Laparoscopic colostomy Parastomal hernia Transperitoneal route

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Methylation-induced silencing of ASC/TMS1, a pro-apoptotic gene, is a late-stage event in colorectal cancer

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CANCER BIOLOGY & THERAPY 2007年第11期6卷 1710-1716页

作者： Riojas, Marco A. Guo, Mingzhou Gloeckner, Sabine C. Machida, emi ota Baylin, Stephen B. Ahuja, Nita Johns Hopkins Univ Dept Surg Baltimore MD USA Johns Hopkins Univ Dept Oncol Baltimore MD USA

The hypermethylation of tumor-suppressor gene promoter regions has been shown to result in the epigenetic inactivation of many genes. ASC/TMS1 is a pro-apoptotic gene that has been shown to be methylated in many different human neoplasms. The methylation status of ASC/TMS1 was analyzed in a series of colorectal cancer (CRC) cell lines, adenomas and primary colorectal cancers and normal colorectal tissue samples using methylation-specific PCR (MSP). The gene expression of ASC/TMS1 in the CRC cell lines was analyzed using reverse-transcriptase PCR (RT-PCR). Methylation analysis showed complete methylation of ASC/TMS1 in 5 of 7 (71%) CRC cell lines. RT-PCR showed absence of mRNA expression in these same cell lines, and expression was restored after treatment with the demethylating drug 5-aza-2'-deoxyazacytidine. The two unmethylated cell lines showed ASC/TMS1 mRNA expression both before and after treatment with 5-aza-2'-deoxyazacytidine. Methylation was seen in 20 of 115 (17%) of primary colorectal cancer specimens, but no methylation was seen in 30 colorectal adenomas and 11 normal colorectal tissue samples. Methylation status of ASC/TMS1 was correlated with a series of clinicopathological variables using multivariate analysis. Methylation of ASC/TMS1 was more common in right-sided tumors (p = 0.02), concordant with hMLH1 methylation (p = 0.03) and is a late stage event, occurring in 0 of 18 tubular adenomas, 0 of 12 villous adenomas, 2 of 44 (5%) Stage 1 cancers, 8 of 31 (26%) Stage 2 cancers, 8 of 21 (38%) Stage 3 cancers and 2 of 19 (11%) Stage 4 cancers. The ASC/TMS 1 gene is frequently silenced in CRC due to promoter hypermethylation. Methylation of ASC/TMS1 appears to be a late-stage event in colorectal carcinogenesis associated with invasive carcinomas but not with normal colorectal tissue or colorectal adenomas. Methylation of ASC/TMS1 may have implications for cancer prognosis.

关键词： ASC/TMS1 methylation colorectal cancer methylation-specific PCR

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Preoperative risk factors for ileostomy-associated kidney injury in colorectal tumor surgery following ileostomy formation

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INTERNATIONAL JOURNAL OF COLORECTAL DISEASE 2024年第1期39卷 1-10页

作者： ota, emi Watanabe, Jun Suwa, Hirokazu Hirai, Tomoya Suwa, Yusuke Nakagawa, Kazuya Ozawa, Mayumi Ishibe, Atsushi Endo, Itaru Yokosuka Kyosai Hosp Dept Surg Yokosuka Japan Kansai Med Univ Dept Colorectal Surg Hirakata Japan Yokohama City Univ Med Ctr Gastroenterol Ctr Dept Surg Yokohama Japan Yokohama City Univ Grad Sch Med Dept Gastroenterol Surg Yokohama Japan

PurposeDiverting ileostomy is related to postoperative high-output stoma (HOS) leading to kidney injury. The purpose of our study was to clarify the risk factors for ileostomy-associated kidney injury, which is kidney injury starting after the first operation to ileostomy closure after colorectal tumor surgery with diverting *** January 2013 and December 2020, 442 patients who underwent colorectal tumor surgery (cancer, neuroendocrine tumor, and leiomyosarcoma) following diverting ileostomy formation were included. We used the KDIGO (Kidney Disease Improving Global Outcomes) guidelines, which defines the acute kidney injury (AKI) to classify patients with ileostomy-associated kidney injury. The definition of AKI was (i) serum creatinine (sCr) >= 0.3 mg/dL or (ii) sCr >= 1.5-fold the preoperative level. Multivariate analyses were performed to identify the independent risk factors for kidney *** injury developed in 99/442 eligible patients (22.4%). Patients in the kidney injury group were older age, male sex, high American Society of Anesthesiologists Physical Status Classification System (ASA-PS) score, hypertension, cardiovascular diseases, diabetes. The preoperative hemoglobin, albumin, prognostic nutritional index (PNI), and creatinine clearance (CCr) were lower, and the maximum wound length was more extended than the non-kidney injury group. The median highest daily stoma output was significantly higher in the kidney injury group. The postoperative white blood cell (WBC) and C-reactive protein (CRP) levels were also high in the kidney injury group. The univariate analysis showed older age, male sex, high ASA-PS score, hypertension, cardiovascular diseases, and diabetes were the risk factors for kidney injury. The multivariate analysis revealed that age 70 or older, ASA-PS III/IV, hypertension, and HOS >= 2000 ml/day were independent risk factors for kidney *** should consider diverting colostomy creatio

关键词： Ileostomy-associated kidney injury Acute kidney injury Colorectal tumor surgery Diverting ileostomy High output stoma

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