ENZYME PATTERNS DURING A SYNCHRONOUS GROWTH CYCLE OF CHLORELLA PYRENOIDOSA

There have been no studies in which a significant number of isozymes have been investigated during a synchronous growth cycle of any organism. The present study was designed to obtain information on the fluctuations of a broad spectrum of enzymes during a synchronous growth cycle of Chlorella pyrenoidosa. Of the thirty-two enzymic activities investigated, seventeen could be localized on starch gels from a high temperature strain of Chlorella pyrenoidosa. Nine of these activities were found to possess more than one band of activity by starch gel electrophoresis. Seven of these activities were localized on starch gels throughout a synchronous growth cycle of C. pyrenoidosa grown in continuous light. Assay techniques are described. The fluctuations in enzyme activity are discussed with relation to concurrent metabolic and cytological changes during cell maturation in C. pyrenoidosa.     

THE PRECISE MEASUREMENT OF HEMOLYSIN

A method is described for the measurement of hemolysin concentration, which makes possible exact comparison of results obtained at different times and with different specimens of erythrocytes and alexin; and gives precise values with an error not greater than 2 per cent.     

低氧时肺动脉内皮细胞单层通透性变化

目的和方法：研究肺动态内皮细胞（ＰＡＥＣ）在低氧性肺水肿（ＨＰＥ）发生中的作用机制。采用ＯＰＡＥＣ体外培养的方法,观察了ＰＡＥＣ生长状态和特征性蛋白因子ⅧＲ：Ａｇ的变化,并利用ＰＡＥＣ融合单层通透性模型研究了低氧对ＰＡＣＥ融合单层的通透性的影响。结果：ＰＡＥＣ生长数量无明显变化,但细胞的生长质量,ⅧＲ：Ａｇ阳性细胞数明显下降,ＰＡＣＥ通透性明显增加,ＣａＭ阻断剂ＴＦＰ只能部分抑制０这种通透性增加。     

CHANGE IN GLUCOSAMINE CONTENT OF CHLORELLA CELLS DURING THE COURSE OF THEIR LIFE CYCLE

1. By the VAN WISSELINGH color reaction and the chitosan sulfatetest it was revealed that Chlorella cells contain chitosan probablyin their cell walls. 2. By fractionating the cell material into several fractionsfollowed by their hydrolysis, it was revealed that the majorityof glucosamine was present in the residue material remaininginsoluble in ethanol-ether and perchloric acid (PCA) solution.Conceivably, this glucosamine has derived, for the most part,from the chitosan contained in the cell wall material. 3. During the course of life cycle of the algal cells, the increasein content of glucosamine occurred in three steps: first, inproportion to the growth of smaller (young) cells into largercells; second, corresponding to the formation of autosporeswithin ripened cells; and third, in parallel with the growthof newly born daughter cells. 4. Between the first and second phase mentioned above, thereoccurred an abrupt breach in the increase of glucosamine. Thisphenomenon was presumed to be closely related to the profoundchange in the permeability of cell walls occuring at this transitionalstage of cell development. (Received September 5, 1960; )     

SYNCHRONOUS CULTURE OF CHLORELLA: I. KINETIC ANALYSIS OF THE LIFE CYCLE OF CHLORELLA ELLIPSOIDEA AS AFFECTED BY CHANGES OF TEMPERATURE AND LIGHT INTENSITY

1. Using the technique of synchronous culture, investigationsweremade of the effects of temperature and light-intensityon cellularlife cycle of Chlorella ellipsoidea. Some improvementsin theculture technique for obtaining a good synchrony of algalgrowthwere described.
2. By following the changes of averagecell volume and cell numberoccurring during culturing, therates of the following processesof life cycle were determined:(i) "growth" (or the increasein cell mass) occurring from thestage of smaller cells (D_a)to the stage of ripened cell (L₃),(ii) "ripening" (or processofformation of "nuclear substances"as estimated from the averagenumber of daughter cells formedfrom single mother cell), and(iii) " maturing and division" which leads to the full maturationof mother cells (L-cells)and their division into separate daughtercells (D-cells).
3. "Growth"and "ripening" were found to be dependent in light,"maturingand division" light-independent. The time requiredfor "growth"and "ripening" (C) is dependent on temperaturebut independentof light intensity, the onset of "maturing anddivision" occurringat the same time (D) of culturing undervaried light intensities.The average cell volume at this stage(L₃),however, was foundto be markedly modified by light intensity;larger with highertemperatures (see Fig. 4).
4. Changes in incubation temperature(under the condition of saturatinglight intensities) were foundto affect the life cycle in thefollowing way: (i) The timeof onset of "maturing and division"(D), varies markedly withculturing temperature; earlier athigher temperatures, (ii)The average cell volume at this stagealso depends on temperature; smaller at higher temperatures.
5. The average number of daughtercells (n) emerging from singlemother cells, was found to beuninfluenced by culturing temperature;(4.0–4.1 underthe conditions of the present study). Itwas found that thedivision number n is remarkably varied bychanging the lightintensity in the "growth" and "ripening"phases; 2.0 at 1 kilolux,3.7 at 5 kilolux, 4.2 at saturatinglight intensities (10 and25 kilolux). This finding was explainedby assuming a light-dependentformation of "nuclear substances"during the "growth" and "ripening"phases, the quantity of thesubstances in the cell at L₃ stagedeterminig the division number.
6. The experimental data wereanalyzed reaction kinetically, therate constants and othercharacteristics of the reactions constitutingthe processesof life cycle were determined, and values forthe apparent activationenergy for each reaction were computed.The reactions were discussedwith special reference to theirrelationship with photosyntheticprocess was discussed.

(Received November 7, 1959; )     

ROLE OF DICTYOSOMES IN WALL FORMATION DURING CELL DIVISION OF CHLORELLA VULGARIS

The behavior of dictyosomes in wall formation during cell division of Chlorella vulgaris follows a definite pattern. During formation of the partition membrane they migrate into the equatorial plane and pair. There is a close spatial relationship between the dictyosomes and the partition membrane which, itself, may be derived from the fusion of dictyosomal vesicles. Dictyosomes also may participate significantly in the deposition of new wall material.     

离体细胞共培养中科间细胞共质体的形成

离体培养下选出的绿色胡萝卜（Ｄａｕｃｕｓ ｃａｒｏｔａ）细胞系和白色普通烟草（Ｎｉｃｏｔｉａｎａｔａｂａｃｕｍ ）细胞系,各自具有独特的细胞结构标志,在愈伤组织、光镜和电镜３ 个水平上均可区分。对两个细胞系进行分散、混合、Ｋ＋ 液低渗处理后在固体培养基上共培养,１０—１５ ｄ 后可观察到两种细胞的镶嵌生长。光镜和电镜下均观察到烟草细胞和胡萝卜细胞之间隔离层的存在与消失。在隔离层消失的区域可见到异种细胞间次生胞间连丝的形成,从而将独立的两个共质体连成一个统一的共质体。对科间细胞共质联系的建立过程进行了讨论,认为细胞接触后首先非特异粘连——以隔离层形成并适度加厚为标志,然后特异的细胞识别在隔离层中启动,从而导致隔离层或消失而重新建立共质联系或加厚、木质化、木栓化     

A STATISTICAL METHOD TO COMPARE THE DEGREE OF MUSCLE CELL MULTIPLICATION IN DIFFERENT CULTURE DISHES

A method was developed for comparing two groups of numbers of cultured muscle cells which were counted under a microscope. Practically important problems for this purpose were: how many fields per dish should be observed, and how many dishes should be prepared under the same conditions, when given test criteria were set.
In the present experiment, 4 dishes were prepared under the same conditions. From each of the dishes, 20 fields were selected, and the numbers of muscle cells *** were counted and separately recorded. Since the purpose was to compare two groups of dishes, the design was a simple case of a nested one. From the experiment, the type of distribution seemed approximately a long-normal distribution with constancy of variance (homoscedasticity). Since the distribution of the cells in dishes belonging to the same group could be considered to be the same, the numbers from each dish could be pooled within a group. Therefore, if the test criteria for Student's t test and the sensitivity to descriminate the ratio of the number of groups are given, the number of fields to be observed per dish times that of dishes can be uniquely determined. This method can be applied for the same purpose to other kinds of cells with log-normal distribution.     

AGE-DEPENDENT CHANGE IN THE TRANSDIFFERENTIATION ABILITY OF CHICK NEURAL RETINA IN CELL CULTURE*

We examined how the transdifferentiation ability of neural retinal cells into lens and/or pigment cells in call culture is changed with the development of the donor. Cells dissociated from neural retinas of chick embryos ranging from 3-day-old to the stage immediately before hatching and of 3-day-old chicks were cultured for about 60 days. The results clearly indicated that the transdifferentiation ability decreased with age. The latest developmental stage at which the differentiation of lens cells took place was in 18-day-old embryos. A gradual decrease in this ability was shown by the comparison of crystallin content in cultures prepared from embryos at different stages. The differentiation of pigment cells was recognized in cultures of neural retinas earlier than in 15-day-old embryos. Such loss of the ability of neural retinal cells to transdifferentiate into pigment cells earlier than into lens cells can be partially attributed to inhibitory factors accumulated in medium conditioned with many neuronal cells present in cultures.     

ROLE OF SULFUR IN THE CELL DIVISION OF CHLORELLA, WITH SPECIAL REFERENCE TO THE SULFUR COMPOUNDS APPEARING DURING THE PROCESS OF CELL DIVISION. I.

1. The role of sulfur in the cell division of Chlorella was studiedby following the fate of the sulfur supplied to the sulfur-deficientcells using ³⁵S as a tracer.
2. The sulfur-deficient cells whichwere unable to perform celldivision were made capable of divisionby the provision of ³⁶S-labeledsulfate under non-photosynthesizingconditions. Soon after theprovision of sulfate the labeledsulfur went rapidly into thecold perchloric acid (PCA)-solublefraction of algal cells,almost entirely in the form of sulfateand/or some other inorganicsulfur substance (s). With the lapseof time, more or less remarkablechanges occurred in the patternof ³⁵S-distribution in differentfractions of cell material.It was noticed that, at the onsetof cell division, a sulfur-containingpeptide-nucleotide compound(s)(SPN), which has been reportedearlier, appeared in a largequantity in the cold PCA-solublefraction, and that its quantitydecreased gradually during thesubsequent process of cell division,suggesting that the compoundwas transformed into some othersubstance (s), presumably withits nucleotide moiety going intonucleic acids and the peptidemoiety going into some essentialproteins.
3. Another noteworthyphenomenon observed during the process ofcell division wasthe incorporation of ³⁶S in a group of hotPCA-soluble substances.These sulfur substances were revealedto be sulfur-containingnucleotidic compounds, which might possiblybe some essentialcomponents of, or substances in close relationto, deoxypentosenucleic acid (DNA).

(Received March 1, 1960; )     

A STATISTICAL APPRAISAL OF THE PROBLEM OF SENSORY MEASUREMENT

The problem of sensory measurement has been tackled using a wide range of methods, from sophisticated systems of scaling to simple ranking. This paper focuses on the problems arising from the fact that the units of sensory scales are frequently arbitrary. It looks at alternative methods of scale design and statistical analysis as a response to the problem and, in particular, introduces a novel combination of scale design and statistical analysis, which may be collectively described as a self-adjusting scale method. The self-adjusting scale method has particular appeal when there is little opportunity for the training of panelists in the use of a particular sensory scale and, though it was originally developed and evaluated in the context of laboratory assessment, may actually have its greatest application in the realm of consumer testing.     

THE DISTRIBUTION OF CELL GENERATION TIMES

Two different distributions of generation times, log-normal and reciprocal normal, were compared by fitting them to experimental generation time distributions for bacteria, yeast, protozoa, and mammalian cells. In every case, the reciprocal normal distribution gave better agreement, whether the original frequency distributions or their cumulatives were compared. the log-normal distribution failed to give significant agreement for three of the original distributions. It is concluded that the reciprocal normal distribution of generation times, or more precisely, the truncated normal distribution of generation rates, is consistent with the published data for steady state populations of all kinds of cells that divide by regular binary fission. A model is suggested for this distribution of generation times, associating it with control of transport of materials into the cell, and with linear cell growth.     

黄斑巨鳖分布的历史变迁

通过考古、文史和动物学文献的考证,对黄斑巨鳖Rafetus swinhoei分布的历史变迁进行了研究,该种历史上曾广泛分布于黄河、长江、太湖、钱塘江、红河、马江流域和台湾澎湖水道,这些区域历史上记录的"鼋"都应为黄斑巨鳖而非亚洲圆鳖Pelochelys cantorii。黄斑巨鳖在台湾澎湖水道仅记录于1万年前,主要是由于人为影响,黄河种群自11世纪后未再有记载,近代其分布区更是急剧缩小,长江中游种群于19世纪末即已绝灭,长江下游和钱塘江种群在20世纪中叶绝灭,太湖流域目前已野生绝灭,红河和马江流域分布区虽未有明显变化,但在最近30年中密度急剧下降,野生种群濒临灭绝。建议重点在红河流域开展研究和保护工作。红河与钱塘江之间是否有分布仍存疑问,需要进一步研究。     

低温诱导小麦叶片细胞表面糖蛋白的变化

利用细胞化学的方法,对经过不同时间低温诱导小麦细胞表面糖蛋白的动态变化进行了标记定位电镜观察。小麦幼苗经２￣３℃下低温锻炼后,细胞表面的糖蛋白层出现明显的增厚,随之开始逐渐地减少变薄,有些细胞表面的糖蛋白层部分脱落于细胞间隙。随低温诱导处理时间不同,细胞表面糖蛋白层表现出的由少量到增厚,最后部分的脱落减少变薄这一有规律的变化现象表明,低温诱导能够引起小麦细胞表面糖蛋白的合成积累,抗寒性较强的冬小麦