日本の山村における地カブの栽培方法について ―静岡県井川地域の事例―

「地カブ」などと呼ばれる自生するカブやカブの仲間については、宮崎県椎葉村の焼畑と関連した事例などの報告がある。しかしこれらは、自生状況の報告などにとどまり具体的な栽培方法は明らかにされていない。そこで本研究では、静岡県井川地域にあるX集落を対象として昭和30（1955）年頃からの山村における地カブ栽培がどのように継承されてきたのかを示し、そこから現地の人にとって地カブの作物としての位置づけと生活文化との関わりを明らかにする。結果は以下のような3点にまとめられる。1つ目は、昭和30（1955）年頃の茶畑および焼畑での地カブ栽培である。X集落における地カブ栽培は、主な栽培地が茶畑と焼畑であった。茶畑では、地カブはチャの株間に自生しており、同じくチャの株間で栽培されていたコンニャクを収穫すると地カブは勝手に生えてきたという。地カブの菜花は養蜂の蜜源に利用した。茶畑の地カブの種子を採集し、焼畑でヒエと地カブを混播した。2つ目は、昭和60（1985）年頃から現在までの常畑における地カブ栽培である。X集落出身のAさんに注目した。Aさんは茶畑だけではなく常畑でもコンニャク栽培を行うようになった。常畑に勝手に生えてきた地カブに花を咲かせ、種子が自然に落ちるよう畑に放置し、X集落の茶畑の地カブ栽培のように、コンニャクを収穫すると地カブが自然と生えてくるような栽培環境を作り出していた。最後は、昭和30（1955）年頃から現在までこぼれ種子を用いる理由についてである。昭和30年代から現在まで地カブの栽培にこぼれ種子を利用することが一貫していた。背景には、地カブは自生力が強いという認識と、自家採種した種子を、いろいろな畑にふりまき、生えてきたものを利用するというAさんの祖母が行っていた栽培方法の影響が考えられた。X集落では、重要な商品作物であったチャやコンニャクを栽培するための茶畑において、コンニャクの収穫が終わると勝手に生える地カブは秋から春先に得られる自給食料として重要ではあるが、あくまで副次的作物として位置づけられて継承されてきた。しかし、焼畑での栽培や養蜂における春先の蜜源としての利用など、山村の生業や生活文化と密接な関わりを持つ作物であったことが明らかとなった。Studies on jikabu, a native turnip species, have been conducted in several areas, including reports about Shiiba Village, Miyazaki Prefecture, where slash-and-burn agriculture had been practiced. However, these reports are fragmentary, and specific cultivation methods have not been clarified. This study aims to clarify the cultivation of native turnips in mountain villages and the transition of cultivation methods from around 1955 to the present. The study site is village X in the Ikawa area of Shizuoka Prefecture. The results are summarized in three points as follows.The first point concerns the cultivation of native turnips around 1955, when slash-and-burn agriculture was still practiced. There were two types of cultivation sites in village X. One was tea fields, where turnips grew naturally between tea plants. Those turnips grew on their own after konjak potato, which was also grown between tea plants, was harvested. Turnip flowers were used as a nectar source for honey bees. The other type of site was slash-and-burn farming fields, where seeds of the turnips collected from the tea fields were planted along with Japanese millet.The second point is the current cultivation method of native turnips in jobata, or permanent agricultural fields since around 1985. Mrs. A from village X began cultivating konjak in a permanent field in addition to a permanent tea field. She let the turnips grow on their own in the permanent field and left their seeds to fall naturally in the field, creating a cultivation environment where turnips would grow naturally after harvesting konjak, just like the turnips in tea fields in village X.The last point refers to the reason for using spilled seeds in the cultivation process. From around 1950 to the present, use of spilled seeds for cultivation of turnips has been common, which may be attributable mainly to the recognition that turnips grow wild and that there was an influence from the cultivation method used since Mrs. A’s grandmother’s generation, in which self-seeded seeds were collected and sowed in various fields.In village X, tea and konjak were important commodity crops. Turnips, which grow on their own in tea fields after konjak is harvested, have been passed down from generation to generation as a by-product, although they are recognized as an important product for consumption by local people from autumn to early spring. This study has also clarified that the locally-grown turnips were closely related to the source of livelihood and the life of the mountain village, including slash-and-burn agriculture and beekeeping

長期ホルマリン固定により失活したProliferating Cell Nuclear Antigen (PCNA) の免疫反応性回復条件の基礎的検討 ―マイクロウェーブ、オートクレーブの影響について―

Using paraffin-embedded tissue sections of liver cancer obtained from autopsy which had been preserved in 10% buffered formalin solution for 6 months while PCNA immunoreactivity was lost, we examined the effects of heat processing by either microwave(MW) and autoclave(AC) in the presence of various processing solution. It appeared that AC processing took shorter time period than MW irradiation to restore equal immunoreactivity. With regard to immunoreactivity retrieval by MW irradiation,however, variation of the degree of retrieval depending on processing time was smaller than in AC, and so the stable consequences were obtained. Although AC processed tissues tended to be stained deep, prolonged processing time presented strong background staining and blurred nuclear margins which made it difficult to estimate the positive cell count. As for the effects of processing solution, there was little difference in retrieval of PCNA among 0.01 M citrate buffer (pH 6.0), saturated solution of lead thiocyanate and distilled water, but the least background staining was observed with distilled water. These observations suggest that MW irradiation of which effect of retrieval is less dependent of processing time and with the least background stainability, is superior to AC processing for PCNA immunoreactivity retrieval on formalin-fixed tissues.10％緩衝ホルマリンに6ヶ月間浸漬していた剖検材料（肝臓癌）のパラフィン包埋後の組織切片を用いて、ホルマリンの固定作用により失活したPCNAの免疫反応性の回復にマイクロウェーブ（MW）及びオートクレーブ（AC）による熱処理とその時用いる処理溶液が、どのような影響を与えるかについて検討した。その結果、同等の免疫反応性を回復するのには、AC処理の方がMW照射より短時間でよいことが解った。しかし、MW照射による免疫反応性回復では、処理時間による影響がAC処理に比べて少なく、安定した結果が得られた。また、PCNAの染色所見については、AC処理の方が濃く染まる傾向が見られた。しかし、処理時間が長くなるとバックグラウンドの染色性が高くなる、核の周囲ににじみ現象が見られる等の所見があり判定に困難をきたした。一方、処理溶液についてはクエン酸（0.01M pH6.0）、チオシアン酸鉛飽和溶液、蒸留水について検討を行った結果、PCNAの回復には差は認められなかったが、バックグラウンドの染色については蒸留水が最も少なかった。以上の結果により、ホルマリン固定により失活したPCNAの免疫反応性を回復するには、MW照射の方がAC処理より処理時間に関係なく安定した染色性が得られ、なおかつバックグラウンドの染色性が少ない等の点で優れていることが示唆された

A new heterozygous compound mutation in the CTSA gene in galactosialidosis

Galactosialidosis is an autosomal recessive lysosomal storage disease caused by the combined deficiency of lysosomal β-galactosidase and neuraminidase due to a defect in the protective protein/cathepsin A. Patients present with various clinical manifestations and are classified into three types according to the age of onset: the early infantile type, the late infantile type, and the juvenile/adult type. We report a Japanese female case of juvenile/adult type galactosialidosis. Clinically, she presented with short stature, coarse facies, angiokeratoma, remarkable action myoclonus, and cerebellar ataxia. The patient was diagnosed with galactosialidosis with confirmation of impaired β-galactosidase and neuraminidase function in cultured skin fibroblasts. Sanger sequencing for CTSA identified a compound heterozygous mutation consisting of NM_00308.3(CTSA):c.746 + 3A>G and c.655-1G>A. Additional analysis of her mother’s DNA sequence indicated that the former mutation originated from her mother, and therefore the latter was estimated to be from the father or was a de novo mutation. Both mutations are considered pathogenic owing to possible splicing abnormalities. One of them (c.655-1G>A) is novel because it has never been reported previously

Quantitative Analysis of Glycosylinositol Phosphoceramide and Phytoceramide 1-Phosphate in Vegetables

Previously, we found an unidentified sphingolipid in cabbage, and determined it as phytoceramide 1-phosphate (PC1P). PC1P is found to be produced from glycosylinositol phosphoceramide (GIPC) by the action of phospholipase D (PLD) activity. Although GIPC is abundant sphingolipid, especially in cruciferous vegetables, amount of daily intake, digestibility and nutritional activity of GIPC are not well understood. Here, we investigated amounts of GIPC and PC1P in vegetables. GIPC was found in all vegetables examined (13 kinds) at levels 3-20mg/100g (wet weight). On the other hand, PC1P was present in limited vegetables which show higher GIPC-PLD activity, such as inner cabbage leaves (5.2mg/100g). Because PC1P is formed during homogenization by activated GIPC-PLD, level of PC1P in boiled cabbage leaves was very low. Although digestibility of GIPC is unknown at present, a portion of dietary GIPC is considered to be converted to PC1P during mastication by plant-derived GIPC-PLD activity in some vegetables

Screening and analysis of edible seaweeds in the ability to adsorb Shiga toxin

We screened edible seaweeds in the ability to adsorb Shiga toxin (Stx) by an equilibrated dialysis method. Although water insoluble fractions of fourteen dry seaweeds did not adsorb Stx, most water soluble fractions were found to adsorb it to one degree or another. Among the seaweed tested, the extract of the Ulva linza Linnaeus [Enteromorpha linza (Linnaeus) J. Agardh] was found to well adsorb both Stx1 and Stx2. We purified the Stx-adsorbing substance from the U. linza extract by DEAE-Toyopearl column chromatography and gel filtration with HiPrep 16/60 Sephacryl S-300 HR column. The purified substance showed an average molecular mass of about 800 kDa by polyacrylamide gel electrophoresis. Analyses of its components indicated that the substance was a highly rhamnose-containing polysaccharide with sulfate esters of 18%. Apparent dissociation constants (Kd) of the polysaccharide to Stx1 and Stx2 were calculated to be 1.9 and 3.5 μM, respectively. To our knowledge, this is the first report on Stx-adsorbing dietary fibers

Transphosphatidylation by GIPC-PLD

Glycosylinositol phosphoceramide (GIPC) is the most abundant sphingolipid in plants and fungi. Recently, we detected GIPC-specific phospholipase D (GIPC-PLD) activity in plants. Here, we found that GIPC-PLD activity in young cabbage leaves catalyzes transphosphatidylation. The available alcohol for this reaction is a primary alcohol with a chain length below C4. Neither secondary alcohol, tertiary alcohol, choline, serine nor glycerol serves as an acceptor for transphosphatidylation of GIPC-PLD. We also found that cabbage GIPC-PLD prefers GIPC containing two sugars. Neither inositol phosphoceramide, mannosylinositol phosphoceramide nor GIPC with three sugar chains served as substrate. GIPC-PLD will become a useful catalyst for modification of polar head group of sphingophospholipid

Targeting critical kinases and anti-apoptotic molecules overcomes steroid resistance in MLL-rearranged leukaemia.

BACKGROUND: Acute lymphoblastic leukaemia with mixed lineage leukaemia gene rearrangement (MLL-ALL) frequently affects infants and is associated with a poor prognosis. Primary refractory and relapsed disease due to resistance to glucocorticoids (GCs) remains a substantial hurdle to improving clinical outcomes. In this study, we aimed to overcome GC resistance of MLL-ALL. METHODS: Using leukaemia patient specimens, we performed bioinformatic analyses to identify target genes/pathways. To test inhibition of target pathways in vivo, we created pre-clinical therapeutic mouse patient-derived xenograft (PDX)-models by transplanting human MLL-ALL leukaemia initiating cells (LIC) into immune-deficient NSG mice. Finally, we conducted B-cell lymphoma-2 (BCL-2) homology domain 3 (BH3) profiling to identify BH3 peptides responsible for treatment resistance in MLL-leukaemia. FINDINGS: Src family kinases (SFKs) and Fms-like tyrosine kinase 3 (FLT3) signaling pathway were over-represented in MLL-ALL cells. PDX-models of infant MLL- ALL recapitulated GC-resistance in vivo but RK-20449, an inhibitor of SFKs and FLT3 eliminated human MLL-ALL cells in vivo, overcoming GC-resistance. Further, we identified BCL-2 dependence as a mechanism of treatment resistance in MLL-ALL through BH3 profiling. Furthermore, MLL-ALL cells resistant to RK-20449 treatment were dependent on the anti-apoptotic BCL-2 protein for their survival. Combined inhibition of SFKs/FLT3 by RK-20449 and of BCL-2 by ABT-199 led to substantial elimination of MLL-ALL cells in vitro and in vivo. Triple treatment combining GCs, RK-20449 and ABT-199 resulted in complete elimination of MLL-ALL cells in vivo. INTERPRETATION: SFKs/FLT3 signaling pathways are promising targets for treatment of treatment-resistant MLL-ALL. Combined inhibition of these kinase pathways and anti-apoptotic BCL-2 successfully eliminated highly resistant MLL-ALL and demonstrated a new treatment strategy for treatment-resistant poor-outcome MLL-ALL. FUNDING: This study was supported by RIKEN (RIKEN President\u27s Discretionary Grant) for FI, Japan Agency for Medical Research and Development (the Basic Science and Platform Technology Program for Innovative Biological Medicine for FI and by NIH CA034196 for LDS. The funders had no role in the study design, data collection, data analysis, interpretation nor writing of the report

Methotrexate Alters the Expression of microRNA in Fibroblast-like Synovial Cells in Rheumatoid Arthritis

We aimed to investigate the effect of methotrexate (MTX) on microRNA modulation in rheumatoid arthritis fibroblast-like synovial cells (RA-FLS). RA-FLS were treated with MTX for 48 h. We then performed miRNA array analysis to investigate differentially expressed miRNAs. Transfection with miR-877-3p precursor and inhibitor were used to investigate the functional role of miR-877-3p in RA-FLS. Gene ontology analysis was used to investigate the cellular processes involving miR-877-3p. The production of cytokines/chemokines was screened by multiplex cytokine/chemokine bead assay and confirmed by ELISA and quantitative real-time PCR. The migratory and proliferative activities of RA-FLS were analyzed by wound healing assay and MKI-67 expression. MTX treatment altered the expression of 13 miRNAs (seven were upregulated and six were downregulated). Among them, quantitative real-time PCR confirmed that miR-877-3p was upregulated in response to MTX (1.79 ± 0.46-fold, p < 0.05). The possible target genes of miR-877-3p in RA-FLS revealed by the microarray analysis were correlated with biological processes. The overexpression of miR-877-3p decreased the production of GM-CSF and CCL3, and the overexpression of miR-877-3p inhibited migratory and proliferative activity. MTX altered the miR-877-3p expression on RA-FLS, and this alteration of miR-877-3p attenuated the abundant production of cytokines/chemokines and proliferative property of RA-FLS