Arsenic Trioxide Sensitizes Glioblastoma to a Myc Inhibitor.

Glioblastoma multiforme (GBM) is associated with high mortality due to infiltrative growth and recurrence. Median survival of the patients is less than 15 months, increasing requirements for new therapies. We found that both arsenic trioxide and 10058F4, an inhibitor of Myc, induced differentiation of cancer stem-like cells (CSC) of GBM and that arsenic trioxide drastically enhanced the anti-proliferative effect of 10058F4 but not apoptotic effects. EGFR-driven genetically engineered GBM mouse model showed that this cooperative effect is higher in EGFRvIII-expressing INK4a/Arf-/- neural stem cells (NSCs) than in control wild type NSCs. In addition, treatment of GBM CSC xenografts with arsenic trioxide and 10058F4 resulted in significant decrease in tumor growth and increased differentiation with concomitant decrease of proneural and mesenchymal GBM CSCs in vivo. Our study was the first to evaluate arsenic trioxide and 10058F4 interaction in GBM CSC differentiation and to assess new opportunities for arsenic trioxide and 10058F4 combination as a promising approach for future differentiation therapy of GBM.滋賀医科大学平成27年

A diverse range of bacterial and eukaryotic chitinases hydrolyzes the LacNAc (Gal<i>β</i>1-4GlcNAc) and LacdiNAc (GalNAc<i>β</i>1-4GlcNAc) motifs found on vertebrate and insect cells

There is emerging evidence that chitinases have additional functions beyond degrading environmental chitin, such as involvement in innate and acquired immune responses, tissue remodeling, fibrosis, and serving as virulence factors of bacterial pathogens. We have recently shown that both the human chitotriosidase and a chitinase from Salmonella enterica serovar Typhimurium hydrolyze LacNAc from Galβ1–4GlcNAcβ-tetramethylrhodamine (LacNAc-TMR (Galβ1–4GlcNAcβ(CH(2))(8)CONH(CH(2))(2)NHCO-TMR)), a fluorescently labeled model substrate for glycans found in mammals. In this study we have examined the binding affinities of the Salmonella chitinase by carbohydrate microarray screening and found that it binds to a range of compounds, including five that contain LacNAc structures. We have further examined the hydrolytic specificity of this enzyme and chitinases from Sodalis glossinidius and Polysphondylium pallidum, which are phylogenetically related to the Salmonella chitinase, as well as unrelated chitinases from Listeria monocytogenes using the fluorescently labeled substrate analogs LacdiNAc-TMR (GalNAcβ1–4GlcNAcβ-TMR), LacNAc-TMR, and LacNAcβ1–6LacNAcβ-TMR. We found that all chitinases examined hydrolyzed LacdiNAc from the TMR aglycone to various degrees, whereas they were less active toward LacNAc-TMR conjugates. LacdiNAc is found in the mammalian glycome and is a common motif in invertebrate glycans. This substrate specificity was evident for chitinases of different phylogenetic origins. Three of the chitinases also hydrolyzed the β1–6 bond in LacNAcβ1–6LacNAcβ-TMR, an activity that is of potential importance in relation to mammalian glycans. The enzymatic affinities for these mammalian-like structures suggest additional functional roles of chitinases beyond chitin hydrolysis

Molecular database for classifying Shorea species (Dipterocarpaceae) and techniques for checking the legitimacy of timber and wood products

The extent of tropical forest has been declining, due to over-exploitation and illegal logging activities. Large quantities of unlawfully extracted timber and other wood products have been exported, mainly to developed countries. As part of the export monitoring effort, we have developed methods for extracting and analyzing DNA from wood products, such as veneers and sawn timbers made from dipterocarps, in order to identify the species from which they originated. We have also developed a chloroplast DNA database for classifying Shorea species, which are both ecologically and commercially important canopy tree species in the forests of Southeast Asia. We are able to determine the candidate species of wood samples, based on DNA sequences and anatomical data. The methods for analyzing DNA from dipterocarp wood products may have strong deterrent effects on international trade of illegitimate dipterocarp products. However, the method for analyzing DNA from wood is not perfect for all wood products and need for more improvement, especially for plywood sample. Consequently, there may be benefits for the conservation of tropical forests in Southeast Asia. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10265-010-0348-z) contains supplementary material, which is available to authorized users

CLICK:One-step generation of conditional knockout mice

Abstract Background CRISPR/Cas9 enables the targeting of genes in zygotes; however, efficient approaches to create loxP-flanked (floxed) alleles remain elusive. Results Here, we show that the electroporation of Cas9, two gRNAs, and long single-stranded DNA (lssDNA) into zygotes, termed CLICK (CRISPR with lssDNA inducing conditional knockout alleles), enables the quick generation of floxed alleles in mice and rats. Conclusions The high efficiency of CLICK provides homozygous knock-ins in oocytes carrying tissue-specific Cre, which allows the one-step generation of conditional knockouts in founder (F0) mice

シンケイ リンパシュショウ デ サイハツシタ チュウスウ シンケイケイ ゲンパツ アクセイ リンパシュ ノ 1ショウレイ

【緒言】神経リンパ腫症は中枢神経または末梢神経への悪性リンパ腫細胞の浸潤によるリンパ節外悪性リンパ腫と考えられる稀な病態である。今回中枢神経系原発悪性リンパ腫の治療後完全寛解後に末梢神経浸潤にて再発を認め、化学療法と自己末梢血幹細胞移植にて完全寛解を得られた症例を経験したので報告する。【症例】50代男性、20XX年に中枢神経系原発悪性リンパ腫と診断、標準的な治療として大量メソトレキセート療法3回施行にて部分寛解、救援療法としてカルボプラチン、エトポシド療法3回と全脳照射(39.6Gy)を施行し完全寛解となった。9ヶ月後、歩行障害、右上肢麻痺、両下肢・右上肢の感覚障害と疼痛を認めた。頭部および脊椎造影MRIに異常所見はなかった。FDG-PET/CTにて右腕神経叢と脊椎神経根に異常集積を認めた。神経リンパ腫症が強く疑われた。脳脊髄液の細胞診とフローサイトメトリー(FCM)にて中枢神経系原発悪性リンパ腫の再発と診断。神経リンパ腫症と診断した。リツキシマブ、メトトレキサート、ビンクリスチン、プロカルバジン併用療法(R-MPV療法)、髄腔内化学療法、およびブスルファン、チオテパ併用(BU/TT)自己末梢血幹細胞移植を施行し完全寛解に至った。【結論】脳脊髄液細胞診とFCM、FDG-PET/CTを併用することで、神経リンパ腫症の診断が可能となった。中枢神経系原発悪性リンパ腫の再発としての末梢神経の神経リンパ腫症ついては確立された治療法はないことを考えると本症例はR-MVP療法とそれに続くBU/TT併用自己末梢血幹細胞移植の有用性を示唆するものである。[Introduction] Neurolymphomatosis is a rare extranodal malignant lymphoma caused by infiltration of malignant lymphoma cells into the peripheral or central nerves. We report a case of neurolymphomatosis as a relapse of primary central nervous system malignant lymphoma. He achieved complete remission with disappearance of neurological symptom by the combination of chemotherapy and autologous peripheral blood stem cell transplantation. [Case] A 50s-year-old male patient was diagnosed as primary central nervous system malignant lymphoma. His disease achieved partial remission with three courses of high-dose methotrexate therapy as induction therapy. The subsequent salvage therapy with carboplatin/etoposide and whole-brain irradiation (39.8Gy) led him complete remission. Nine months later, he developed gait disorder, right upper extremity paralysis, sensory disturbance and pain in both lower extremities and upper right extremities. There were no abnormal findings on head and spine imaging MRI. However, PET-CT scan demonstrated abnormal uptake of FDG in the right brachial plexus and spinal nerve roots. The cerebrospinal fluid cytology and flow cytometry confirmed the diagnosis of neurolymphomatosis as a recurrence of primary central nervous system malignant lymphoma. He underwent R-MPV therapy, intrathecal chemotherapy, and autologous peripheral blood stem cell transplantation with a preconditioning of busulfan and thiotepa, resulting in complete remission of neurolymphomatosis with disappearance of neurological symptoms. [Conclusion] The cerebrospinal fluid cytology and flow cytometry together with PET-CT enabled us to make a diagnosis of neurolymphomatosis. Given that there are no established treatments for neurolymphomatosis of the peripheral nerves as a recurrence of primary central nervous system malignant lymphoma, this case suggests the possible efficacy of the R-MPV therapy followed by autologous peripheral blood stem cell transplantation with busulfan and thiotepa

Cerebral aneurysms and inflammation

Multiple inflammatory factors, playing a crucial role in cerebral aneurysm formation, have been identified. Tumor necrosis factor-alpha (TNF-α) has been revealed to have a close connection with several risk factors that affect aneurysm formation. Remarkable expression in aneurysm walls of mRNA for TNF-α has been observed in humans. Possible therapeutic interventions to reduce the formation of cerebral aneurysms may include the inhibition of mediators of inflammation

The mRNA Expression of Neurotrophins in Different Skeletal Muscles of Young Rats

Skeletal muscles are a target for motoneurons and synthesize neurotrophins, including nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4/5 (NT-4/5). Both at the embryonic stage and the adult stage, the mRNA expression of neurotrophins in skeletal muscles of rats has been reported. However, little was known about the mRNA expression patterns of neurotrophins in skeletal muscles of rats at the young developmental stage. In this study, we investigated the mRNA expressions of BDNF and NT-3 in three different skeletal muscles in 4 - to 8 - week - old rats using the reverse transcriptional polymerase chain reaction (RT-PCR) method. The expression of BDNF mRNA in the soleus muscle gradually became higher with age from 5 to 8 weeks. But BDNF mRNA in the tibialis anterior and extensor digitorum longus muscles did not change with growth. The expression of NT-3 mRNA did not show a specific tendency during this period. The differences of muscle fiber types, recruitment patterns of the muscles, and roles of neurotrophins may cause these mRNA expression patterns. Neurotrophins are target-derived, activity-dependent neurotrophic factors and are transported retrogradely. There is a possibility that the different expression patterns of neurotrophins in muscles may be involved in the maturation of neuromuscular function in different muscles during the young developmental period