CHROMATOGRAPHIC ANALYSES OF ACID-SOLUBLE POLYPHOSPHATES IN CHLORELLA CELLS

Using uniformly ³²P-labeled Chlorella cells as material, compositionof acid-soluble inorganic polyphosphates was studied by paperchromatography and ion-exchange chromatography. 2.By the paper chromatographic analysis it was found that theacid-soluble polyphosphates consisted of highly condensed polyphosphates.Ring-forming tri- and tetrametaphosphates, pyrophosphate andtripolyphosphate were not detected in the acid-soluble fractionof the algal cells. 3.By an ion-exchange chromatography with the use of increasingconcentrations of KCl-solution as eluant, it was found thatthe acid-soluble polyphosphate was a mixture of polyphosphateswith a variety of condensation number (n-values). Polyphosphatesof the n-values between 3 and 15 were only 20% of the totalacid-soluble polyphosphate. The majority of the other polyphospateshad greater n-values which was eluted with 0.5–1.0 M KCl. (Received March 2, 1964; )     

Effect of Gossypol on Trypomastigotes and Amastigotes of Trypanosoma cruzi

Bloodstream Trypanosoma cruzi trypomastigotes isolated from infected mice undergo reduction of motility and structural damages after 5 to 45 min exposure to gossypol at concentrations ranging from 5 to 50 μM. When 1% serum albumin is added to the incubation medium, no alterations of parasites are observed, even with 100 μM gossypol. Intracellular T. cruzi amastigotes in infected Vero cell cultures exposed to 5 μM gossypol for 2 h do not show changes. Incubation with 5 μM gossypol for 48 h produces complete disruption of host cells; however, the amastigotes they contain show only mineor alterations. The observations indicate that, in protein-rich media, gossypol is complexed into associations which have no activity on the different forms of the T. cruzi biological cycle.     

Isolation of tobacco mesophyll cells in intact and active state

A procedure using a fungal pectinase was developed to rapidlyrelease mesophyll cells from tobacco leaves. Fifty to ninetyper cent of the released cells were morphologically intact andwere converted into spherical protoplasts by cellulase treatment.Cells isolated from tobacco mosaic virus-inoculated leaves supportedmultiplication of the virus during subsequent incubation. (Received December 14, 1967; )     

High temperature causes arrest of cell cycle in G2 phase in BmN cells derived from the silkworm, Bombyx mori

Abstract.  The influence of temperature on the insect cell line, BmN, derived from the silkworm, Bombyx mori is investigated. These cells proliferate at an accelerated pace as the temperature increases from 22 to 30 °C, but the growth rate slows at 34 °C, and proliferation stops at 38 °C. At high temperatures, abnormal cellular morphology is observed. Cells treated at 38 °C have cytoplasmic bilateral protrusions and they gradually aggregate and float in the medium. BmN cells without proliferation at 38 °C are viable but have reduced DNA synthesis. At high temperatures, the cell cycle of BmN cells halts at the G₂ phase. After heat treatment of the larvae, an accumulation of larval haemocytes with high DNA content is found, which suggests that the cell cycle arrest at G₂ also occurs in the silkworm at high temperatures.     

低剂量率下高线性能量转移碳离子束辐照人类唾液腺细胞的存活效应

利用日本千叶重离子医用加速器 HIMAC 提供的碳离子束,对人类唾液腺细胞 (HSG) 在剂量率为 0.5 Gy/h 的低剂量率条件下进行了辐照,运用标准的克隆形成法得到了 3 种不同剂量平均线性能量转移 (LET) 碳离子束辐照 HSG 细胞的剂量存活效应 . 与先前 HSG 细胞在治癌剂量率 (1~5 Gy/min) 下对相近剂量平均 LET 碳离子束辐照的剂量存活效应数据相比, HSG 细胞对高 LET 碳离子束辐射表现出明显的剂量率效应 . 为在相同条件下得到碳离子束对 HSG 细胞的相对生物学效应 (RBE) ,利用 60Co-γ射线在剂量率为 0.5 Gy/h 的条件下辐照了 HSG 细胞,得到该细胞系对低 LET 射线响应的剂量存活效应 . 与先前在治癌剂量率下得到的 RBE 值相比,低剂量率条件下得到的 RBE 值总体减小 . 由实验发现的剂量率效应及低剂量率条件下 RBE 值的减小,表明由高 LET 碳离子束造成的辐射损伤在低剂量率条件下也存在着显著的修复效应 . 据此,对辐射造成细胞致死的原因进行了探讨 .     

QUANTITATIVE SEPARATION OF INORGANIC POLYPHOSPHATES IN CHLORELLA CELLS

Using unicellular green alga, Chlorella ellipsoidea, which hadbeen labeled with ³²P-phosphate, re-examination was performedon our routine method for the quantitative separation of inorganicpolyphosphates (poly-Pi). Confirming the previous results, itwas demonstrated that poly-Pi's in Chlorella cells are clearlyseparated from each other by successive extractions with cold8% TCA (poly-Pi "A"), with cold KOH at pH 9 (poly-Pi "B"), andwith 2 N KOH (poly-Pi's "C" and "D"; the former precipitateson neutralization with PCA, leaving the latter in solution).Use of 2N KOH was found to be most suitable for the purposeof fractionation of poly-Pi's "C" and "D." (Received February 25, 1965; )     

Efficiency of Countercurrent Distribution, Sephadex G-10, and Silicic Acid Partition Chromatography in the Purification and Separation of Gibberellin-like Substances from Plant Tissue

The effectiveness of countercurrent distribution, Sephadex G-10column chromatography and silicic acid partition column chromatographyin the purification of gibberellin-like substances from extractsof etiolated Phaseoltu multiflorus seedlings, and elongatingvegetative shoots of Arizona cypress (Cupressus arizonica Greene)and coastal Douglas-fir (Pseudotsuga menziesii var. menziesii)was followed by the use of the barley half-seed -amylase bioassayand dry-weight measurements. Countercurrent distribution resultedin a 10- to 30-fold reduction in the dry weight of the acidic,ethyl acetate-soluble fraction. Sephadex G-10 column chromatographyfurther lowered the dry weight by about two-thirds. Silicicacid partition column chromatography separated gibberellin-likesubstances from each other and again reduced the dry weight.Enhancement of bioassay activity was noted at each step of thepurification procedure. It is concluded that the above proceduresconstitute useful and efficient tools for the initial purificationof gibberellin-like substances from plant tissue.