Amyloidosis is often overlooked because its clinical manifestations can mimic those of more-common diseases. It is important to get a precise diagnosis as early as possible for the prevention of further organ damages. Amyloidosis is a disorder caused by deposition of insoluble abnormal amyloid. The kidney is a frequent site of amyloid deposition. The amyloid fibrils have a characteristic appearance and generate birefringence under polarized light when stained with the Congo red dye. Classification of amyloidosis is based on the precursor protein that forms the amyloid fibrils and the distribution of amyloid deposits as either systemic or localized. Involvement of amyloid fibrils in kidneys mainly occurs as amyloid light-chain (AL) or amyloid A (AA) amyloidosis. The potassium permanganate method with Congo red staining was once used widely to discriminate AL and AA amyloidoses, but this method has a problem of false positive results. We found that extracellular and cytoplasmic glomerular 4', 6-diamidino-2-phenylindole (

)-positive areas were clearly consistent with amyloid deposition in AL amyloidosis. In contrast, the overlapping staining was not seen in AA amyloidosis. Therefore, we propose that staining readily distinguishes AL renal amyloidosis from AA renal amyloidosis as a simple and reproducible histochemical method. J. Med. Invest. 64: 217-221, August, 2017

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モモの染色体(2n=16)をCMA(chromomycin A₃)および(4'-6-diamidino-2-phenylindole)により染色し, その蛍光分染パターンから染色体の識別を試みた.'おはつもも', 'あかつき'および'モモ台木筑波4号'の自然交雑実生の根端を材料とした.根端は2mMの8-オキシキノリン液で10℃4時間の前処理を行い, 酵素解離法および風乾法により染色体標本を作成した.CMAおよび染色を行うと16本のうち6本(3対)の染色体でCMA+/-バンドが観察された.これらのバンドは2本の染色体の付随体部分, 2本の染色体の端部, 2本の染色体の基部に出現した.CMA+/-バンド数および染色体上での位置は安定しており, 品種間および個体間での差異はなかった.また, CMA-/+バンドは認められなかった.

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クロタネソウNigella damascena L. の間期核と体細胞分裂中期染色体をCバンド処理後、クロモマイシンA3(CMA)とで染色を行った。すべての染色体の動原体部にある対をなすCドットは染色で蛍光を発し、3対の中部動原体型染色体の二次狭窄部のCバンドはCMA染色で蛍光を発した。残りのCバンドもCMAあるいは染色で、蛍光を発した。

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Bacterial abundance determined with SYBR Green I and 4'6-diamidino-2-phenylindole () methods were compared using the protocol for each method employing epifluorescence microscopy. This comparison, using seawater samples that were collected from Sagami Bay, revealed that the abundance determined with the SYBR Green I method was on average 1.6 fold greater than that with the method. A model 2 regression analysis showed the following relationship:
Log SYBR Green I=-0.238+1.08 Log (r²=0.85, n=68, p<0.0001). These results suggest that when the method is switched between the SYBR Green I and methods, the difference in bacterial abundance determined by the two methods should be considered for a quantitatively consistent assessment of the bacterial biomass in the ocean.

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Objective: This study examined the adhesive properties and trial manufacture of resin core shen it is applied to the infected root canal wall. Materials and Methods: The examined resins were: AC-1, AC-2 and AC-3, which contain an acid reaction fluoride inclusion of glass ionomer filler (S-PRG). Evaluations of bonding strength and bonding elasticity were performed. The flow value was calculated and an adhesion test of the resins was performed according to the ISO standard. The bacterial survivability on the root canal wall was analyzed by scanning electron microscope (SEM) and 4', 6-diamino-2-phenylindole dihydrochloride (). Results: A bacterial survivability on the root canal wall of 76% was confirmed by SEM and . In addition, a rate of concordance between anaerobiotic culture and of 86% was exhibited. However, neither infiltration nor pigmentation was observed between the root canal wall and resin core. Suitable seal and absence of gap was corroborated by SEM. The elemental components of the bonding materials were identified by energy dispersive X-ray micro analyzer. Conclusions: The simple confirmation of bacterial survivability by was effective and the trial manufacture of root canal resin core presented favorable behavior in infected root canal wall with physical properties similar to the dentin. Therefore, the clinical application of these materials could be possible.

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マウスに薬剤感受性の異なるTrypanosoma congolenseを感染させ, diminazene aceturateまたはisometamidium chlorideを投薬することにより, 末梢血中に出現するkinetoplasのない(AK)型の観察に染色を用い, ギムザ染色と比較した. 投薬10時間後のAK型発現率を求める観察には両染色法に差はなかったが, 正常のトリパノソーマとAK型のそれとの区別する観察には, 染色がギムザ染色に比べ優れていた.

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Karyotype analysis of three improved grass pea (Lathyrus sativus L.) varieties, namely Nirmal, BioL-203 and BioR-231 was carried out using root tip metaphase cells after staining with orcein and with two DNA base specific fluorochromes, Chromomycin A3 (CMA) and 4-6'diamidino 2-phenyle indole (). It was revealed that the three varieties were diploids showing 2n=14 with metacentric and submetacentric chromosomes. The three varieties showed minor variations in conventional orcein staining, but were distinctly different from each other in GC-specific CMA and AT-specific -banding patterns. CMA-stained regions fluoresced as bright bands mainly in terminal secondary constriction and peri-centromeric regions, while -stained parts were highlighted both as bands and pairs of dots in terminal regions of different pairs. The study revealed conspicuous differences among seven pairs of chromosomes and presence of one pair of chromosomes with secondary constriction, pair 1 in BioR-231 and pair 2 in both BioL-203 and Nirmal, containing GC-rich repeats. Among the varieties, Nirmal contained the highest percentage of GC-rich sequences exhibiting 12 CMA-positive bands, and it was followed by BioL-203 (10 bands) and BioR-231(eight bands). In -banding, the highest number of bands (12) was scored in BioL-203, followed by BioR-231 (10) and Nirmal (eight). The variation in banding patterns suggests the presence of karyomorphological polymorphism in the present materials.

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The distribution of mollicutes in periwinkle plants (Catharanthus roseus (L.) G. Don) that had been graft-inoculated with the clover proliferation (CP) agent or the potato witches'-broom (PWB) agent was studied by in situ molecular hybridization with biotinylated single-stranded DNA probes and by DNA staining of hand sections of petioles and stem internode tissues with (4', 6-diamidino-2-phenylindole). In situ hybridization with the biotinylated probes was specific, detecting only the CP and PWB agents but not the western aster yellows (AY27) agent, which was detected by . The mollicutes were detected first in the external primary phloem tissue and then in the recently infected secondary phloem elements. Later, the spread of mollicutes was detected in the internal phloem tissue. In situ hybridization was highly specific and sensitive in detecting, without background signals, mollicutes in young sieve elements. However, the complete absence of hybridization signals and of DNA staining with coincided with degeneration of affected sieve tubes in the primary phloem elements.

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The chromosomes and interphase nuclei of Genipa americana L. were analyzed after CMA/ staining. Most of the large chromocentres was CMA+/+, except that associated with the nucleolus which was CMA + + / - -, whilst the diffuse euchromatin was CMA-/+. In addition, some small chromocentres were CMA-/+. The chromosome bands were CMA + / + or CMA + +/- -. Most of the chromosome pairs showed size heteromorphism, mainly in the CMA+/+ bands. Measurements of the heterochromatin/euchromatin ratio in metaphase chromosomes gave an estimate of about 50%, one of the highest ratio observed in angiosperms. Comparison with some other species with a high heterochromatin amount suggest that heterochromatin accumulation may be an aleatory rather than an adaptive process.

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Karyotype analysis of 3 released varieties in Vigna radiata L. viz. Barimung-2, Barimung-3, Barimung-5 has been carried out after staining with orcein, CMA and . These varieties were found to possess 2n=22 chromosomes. Four terminal CMA-positive bands were found in Barimung-2, 3 such bands in Barimung-3 and 4 in Barimung-5. The terminal CMA positive bands showed deep -negative reversible bands in the 3 varieties of V. radiata. One chromosome of Barimung-5 was fluoresced entirely with CMA and . -positive and -negative band occurred in another chromosome of Barimung-5. A pair of interstitial CMA-negative band was observed in chromosome pair V of only Barimung-2. These portions were stained brightly with . These kinds of reversible banded chromosomes could be easily identified. Polymorphism regarding the number and location of -positive bands were observed in these varieties. That may due to either minute deletion or high condensation of heterochromatic region at the respective loci. Few chromosomes of each variety showed characteristic CMA and bands. Therefore, with the help of CMA and it was possible to develop marker chromosomes for authentic identification of three varieties in V. radiata.

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Fluorescent vital staining and rRNA-targeted in situ hybridization were used to investigate the physiological activity and microbial population structures of two types of activated sludge: fresh returned sludge from a municipal wastewater treatment plant and MITI-standard sludge (a microbial source in a biodegradation test, Modified MITI Test). The ratio of esterase-active bacteria, as determined by double-staining with 4', 6-diamidino-2-phenylindole () and 6-carboxyfluorescein diacetate (6CFDA), was 60-80% to total bacterial number in both types of activated sludge; both ratios being much higher than those of the colony forming bacteria. Microbial population structures in both types of activated sludge were determined by 2-hydroxy-3-naphthoic acid-2'-phenylanilide phosphate (HNPP) and Fast Red TR in situ hybridization (HNPP-FISH). Approximately 50% of the -stained bacteria was detected by HNPP-FISH in wastewater treatment plant sludge and 40% in the MITI-standard sludge. The bacterial community structure in wastewater treatment plant sludge was dominated by the Flavobacterium-Cytophaga group. Characteristic coccoid cells, which fluoresced bright yellow under UV excitation after -staining, were dominant in the MITI-standard sludge.

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Correspondence of banding patterns of Giemsa C-band and /CMA fluorochromes of the same specimen of Aconitum sanyoense was examined. It appeared that there were various C-bands, that is, AT-rich (C⁺/+/CMA^-), GC-rich (C⁺/-/CMA⁺), and intermediate (C⁺/-/CMA^-) C-bands. Furthermore, some so-called euchromatic segments showed AT-rich DNA sequences (C^-/+/CMA^-). Determination of the combination of these banding patterns may advance detailed detection of cytogenetical variation among individuals of this species.

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　 本研究の目的は，細菌のDNAを蛍光染色する色素４ ’，6-diamidino-2-phenylindole（）を用い，細菌の存在を簡便に可視化することによって，口腔内細菌数とその変動を調査し，口腔ケアの評価として活用の可能性を検討することである。同意の得られた健康な成人10名と高齢者10名を対象に，口腔内細菌の日内変化と口腔内状況や生活習慣を調査した。さらに，Ａ看護大学の学生11名を対象に，口腔ケア演習の前後の口腔内細菌数を調べた。細菌は，口腔内３か所（舌の上，頬の内側，上下歯の表面）を清潔な綿棒で拭って採取した。結果，は短時間に検体中の細菌の存在を可視化することができ，さらに数を計測することが可能となり，口腔ケアの評価として活用できることが明らかとなった。

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カラタチ (Poncirus trifoliata (L.) Raf.) 染色体のクロモマイシン A₃ (CMA) 染色およびプロピジウムイオダイド (PI)/4'-6-ジアミジノ-2-フェニルインドール () 染色を行った．PI/ 染色の PI (+) 領域は CMA 染色の CMA (+) 領域と一致しており，PI (+) シグナルは CMA (+) シグナルに比べ安定していた．PI (+) シグナルの相対的な大きさを加味したバンドパターンとシグナルのない染色体の形態から，18 本のカラタチ染色体は 9 グループ (2B₁ + B₂ + B₃ + 4D₁ + 2D₂ + D₃ + D₄ + 2E_L + 4E) に分けられた．ディゴキシゲニン-ローダミン（赤色）でラベルしたカラタチDNAとビオチン-FITC（緑色）でラベルしたウンシュウミカン‘南柑 20 号’(Citrus unshiu Marcow.) または‘土佐文旦’(Citrus maxima (Burm.) Merr.) DNA の二重プローブを用いてカラタチ染色体に genomic in situ hybridization (GISH) を行った．明瞭な GISH シグナルが PI (+) 領域と同じ位置に検出された．GISH シグナルの色調はビオチンプローブをウンシュウミカンとしたときは黄色と緑色，‘土佐文旦’としたときは黄色と赤色であった．このことから，カラタチは‘土佐文旦’よりウンシュウミカンに近縁であると考えられた．すべての B 型染色体の二次狭窄部と D₄ 型染色体端部の GISH シグナルは，いずれのビオチンプローブでも黄色または緑色であったことから，カラタチ染色体のこの部位はウンシュウミカンおよび‘土佐文旦’ゲノムと相同性が高く，保存領域である可能性が示唆された．

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