DNA markers based on retrotransposon insertion polymorphisms can detect short DNA fragments for strawberry cultivar identification

In this study, DNA markers were developed for discrimination of strawberry (Fragaria × ananassa L.) cultivars based on retrotransposon insertion polymorphisms. We performed a comprehensive genomic search to identify retrotransposon insertion sites and subsequently selected one retrotransposon family, designated CL3, which provided reliable discrimination among strawberry cultivars. Through analyses of 75 strawberry cultivars, we developed eight cultivar-specific markers based on CL3 retrotransposon insertion sites. Used in combination with 10 additional polymorphic markers, we differentiated 35 strawberry cultivars commonly cultivated in Japan. In addition, we demonstrated that the retrotransposon-based markers were effective for PCR detection of DNA extracted from processed food materials, whereas a SSR marker was ineffective. These results indicated that the retrotransposon-based markers are useful for cultivar discrimination for processed food products, such as jams, in which DNA may be fragmented or degraded

Efficient screening of long terminal repeat retrotransposons that show high insertion polymorphism via high-throughput sequencing of the primer binding site

Retrotransposons have been used frequently for the development of molecular markers by using their insertion polymorphisms among cultivars, because multiple copies of these elements are dispersed throughout the genome and inserted copies are inherited genetically. Although a large number of long terminal repeat (LTR) retrotransposon families exist in the higher eukaryotic genomes, the identification of families that show high insertion polymorphism has been challenging. Here, we performed an efficient screening of these retrotransposon families using an Illumina HiSeq2000 sequencing platform with comprehensive LTR library construction based on the primer binding site (PBS), which is located adjacent to the 5′ LTR and has a motif that is universal and conserved among LTR retrotransposon families. The paired-end sequencing library of the fragments containing a large number of LTR sequences and their insertion sites was sequenced for seven strawberry (Fragaria × ananassa Duchesne) cultivars and one diploid wild species (Fragaria vesca L.). Among them, we screened 24 families with a “unique” insertion site that appeared only in one cultivar and not in any others, assuming that this type of insertion should have occurred quite recently. Finally, we confirmed experimentally the selected LTR families showed high insertion polymorphisms among closely related cultivars

A case of primary renal angiosarcoma

A 78-year old man was diagnosed with a left bleeding renal cyst from CT scan results. Serial CT scans revealed the left kidney mass to be increasing in size and a new lesion in the liver. Renal cell carcinoma with liver metastasis was diagnosed and a radical nephrectomy performed. The initial pathological diagnosis was a benign chronic hematoma. However, the liver mass increased in size and multiplied, while another mass emerged in the twelfth thoracic vertebra with spinal paralysis and was immediately removed. Pathological findings for that specimen showed malignancy of stromal cell origin but low atypia. The renal specimen was re-evaluated using whole cross-section analysis and immunohistochemistry, and diagnosed as a primary renal angiosarcoma. Recombinant interleukin-2 therapy was started immediately; however, the patient died of metastatic disease 13 months after the initial operation. Although contrast imaging depicted the primary lesion as a non-specific hematoma with little focal pooling, and low-grade cytological atypia was shown pathologically, the angiosarcoma was extremely aggressive

前立腺癌患者における骨転移および続発性骨粗鬆症診断に対する尿中NTX測定の有用性

〔目的〕骨代謝マーカーであるtype I collagen cross-linked N-telopeptides(NTX)の尿中濃度は骨粗鬆症,副甲状腺機能亢進症および悪性腫瘍の骨転移診断に有用であると言われている.今回我々は前立腺患者の尿中NTXを測定し,骨転移やホルモン療法による続発性骨粗鬆症の診断に対する有用性について検討を行った.〔対象と方法〕前立腺癌と診断された51症例に対して骨代謝マーカーである尿中NTX,血中pyridinoline cross-linked carboxyterminal telopeptide of type I collagen(ICTP),血中carboxyterminal propeptide of type I procollagen(PIPC),血中bone specific alkaline phosphatase(BAP)の測定を行い,それぞれのマーカーとの相関,ホルモン療法および骨転移との関係について検討した.〔結果〕ホルモン療法の期間と尿中NTXには有意な相関(p=0.008)を認めた.治療法に従って2群に分け,各骨代謝マーカーについて検討した.尿中NTXはホルモン療法群で非ホルモン療法群に比して有意に高値を示した.骨転移の有無で2群に分け,各骨代謝マーカーについて検討した.2群間で各骨代謝マーカーに有意差は認められなかった.〔結論〕ホルモン療法の期間と尿中NTXに相関が認められた.長期ホルモン療法の患者では尿中NTXの定期的な測定により,早期の続発性骨粗鬆症の診断および治療に有用であると思われた.[Aim] A high urinary level of type I collagen cross-linked N-telopeptides (NTX), a bone metabolism marker, is associated with osteoporosis, hyperparathyroidism, and bone metastasis in patients with malignant tumors. We investigated the significance and usefulness of measuring the urinary NTX concentration for detection of bone metastasis (BM) and secondary osteoporosis due to hormone therapy (HT) in prostate cancer patients. [Methods] The levels of four bone metabolism markers, urinary NTX, blood ICTP, blood PIPC, and blood BAP, in 51 patients with prostate cancer, were compared with whether patients received HT and the status of bone metastasis. [Results] The duration of HT was significantly correlated with urinary NTX concentration (p=0.008). The urinary NTX concentration was significantly higher in patients who received HT than in patients who did not. There were no significant differences in the levels of the four metabolism markers between patients who did or did not have bone metastasis. [Conclusion] The duration of HT was correlated with the urinary NTX concentration. In patients who have received long-term HT, periodical measurement of urinary NTX level is considered to be useful for early detection and treatment of secondary osteoporosis

前立腺癌患者における骨転移および続発性骨粗鬆症診断に対する尿中NTX測定の有用性

〔目的〕骨代謝マーカーであるtype I collagen cross-linked N-telopeptides(NTX)の尿中濃度は骨粗鬆症,副甲状腺機能亢進症および悪性腫瘍の骨転移診断に有用であると言われている.今回我々は前立腺患者の尿中NTXを測定し,骨転移やホルモン療法による続発性骨粗鬆症の診断に対する有用性について検討を行った.〔対象と方法〕前立腺癌と診断された51症例に対して骨代謝マーカーである尿中NTX,血中pyridinoline cross-linked carboxyterminal telopeptide of type I collagen(ICTP),血中carboxyterminal propeptide of type I procollagen(PIPC),血中bone specific alkaline phosphatase(BAP)の測定を行い,それぞれのマーカーとの相関,ホルモン療法および骨転移との関係について検討した.〔結果〕ホルモン療法の期間と尿中NTXには有意な相関(p=0.008)を認めた.治療法に従って2群に分け,各骨代謝マーカーについて検討した.尿中NTXはホルモン療法群で非ホルモン療法群に比して有意に高値を示した.骨転移の有無で2群に分け,各骨代謝マーカーについて検討した.2群間で各骨代謝マーカーに有意差は認められなかった.〔結論〕ホルモン療法の期間と尿中NTXに相関が認められた.長期ホルモン療法の患者では尿中NTXの定期的な測定により,早期の続発性骨粗鬆症の診断および治療に有用であると思われた.[Aim] A high urinary level of type I collagen cross-linked N-telopeptides (NTX), a bone metabolism marker, is associated with osteoporosis, hyperparathyroidism, and bone metastasis in patients with malignant tumors. We investigated the significance and usefulness of measuring the urinary NTX concentration for detection of bone metastasis (BM) and secondary osteoporosis due to hormone therapy (HT) in prostate cancer patients. [Methods] The levels of four bone metabolism markers, urinary NTX, blood ICTP, blood PIPC, and blood BAP, in 51 patients with prostate cancer, were compared with whether patients received HT and the status of bone metastasis. [Results] The duration of HT was significantly correlated with urinary NTX concentration (p=0.008). The urinary NTX concentration was significantly higher in patients who received HT than in patients who did not. There were no significant differences in the levels of the four metabolism markers between patients who did or did not have bone metastasis. [Conclusion] The duration of HT was correlated with the urinary NTX concentration. In patients who have received long-term HT, periodical measurement of urinary NTX level is considered to be useful for early detection and treatment of secondary osteoporosis