A CHROMATOGRAPHIC STUDY OF HYDROLYSIS IN THE FEULGEN NUCLEAL REACTION

In a study on Feulgen hydrolysis of frozen-dried alcohol-fixed lily anthers, a chromatographic technique was developed to analyze the acid hydrolysate for some of the degradation products of nucleic acid. Hydrolysis was accomplished by 10 per cent perchloric acid at 20°C., and a typical hydrolysis time-Feulgen intensity curve was obtained, with maximum staining occurring at 19 hours. Microphotometric measurements indicated that the amount of stain per nucleus was no different from amount in nuclei fixed and hydrolyzed by more conventional procedures. Uracil-containing material (from ribonucleic acid) was almost completely separated from thymine-containing material (deoxyribonucleic acid) of tissue sections by acid treatment for 1½ hours. Adenine (purines), as the base, was effectively all removed from the deoxyribonucleic acid at the time of optimum hydrolysis. Detectable amounts of thymine-containing material appeared in the hydrolysate shortly after the onset of hydrolysis; and the amount increased rapidly with increased hydrolysis time. At the time of optimum hydrolysis approximately two-thirds of the total deoxyribonucleic acid thymine was lost. The removal of these thymine-containing fragments was linear with respect to time during the first 24 hours and occurred at a relatively high rate. Removal after 24 hours was also linear but was at a markedly lower rate. These results would suggest that two kinds of deoxyribonucleic acid exist in lily anthers; an acid-labile fraction amounting to approximately three-fourths of the total, and an acid-resistant fraction making up the remainder. In the Feulgen procedure much of the labile fraction is lost by the time of optimum hydrolysis and is not stained; most of the stable fraction remains in the tissue and is stained. In light of these findings the use of the Feulgen method as a means of determining cytochemically relative amounts of deoxyribonucleic acid in nuclei by measuring their Feulgen dye content was discussed.     

QUANTITATIVE GENETICS AND THE EVOLUTION OF REACTION NORMS

We extend methods of quantitative genetics to studies of the evolution of reaction norms defined over continuous environments. Our models consider both spatial variation (hard and soft selection) and temporal variation (within a generation and between generations). These different forms of environmental variation can produce different evolutionary trajectories even when they favor the same optimal reaction norm. When genetic constraints limit the types of evolutionary changes available to a reaction norm, different forms of environmental variation can also produce different evolutionary equilibria. The methods and models presented here provide a framework in which empiricists may determine whether a reaction norm is optimal and, if it is not, to evaluate hypotheses for why it is not.     

MICROSPECTROPHOTOMETRY OF CELL NUCLEI STAINED WITH THE FEULGEN REACTION : IV. FORMATION OF TETRAPLOID NUCLEI IN RAT LIVER CELLS DURING POSTNATAL GROWTH

1. DNA contents of the individual parenchymal nuclei of rat livers during postnatal growth were estimated by microspectrophotometric apparatus, and different ploidy classes of nuclei were classified by their DNA contents. With the same material the total number of parenchymal nuclei in the liver was counted microscopically. 2. If the DNA content of nuclei encountered most frequently in several tissues represents the diploid class, the ploidy classes of the rat liver cell nuclei correspond to di-, tri-, tetra-, and octoploid, with the di- and tetraploid ones predominating considerably. 3. In suckling rats (below 25 gm. of body weight) the liver parenchyma is composed almost exclusively of cells with diploid nuclei, whereas in young rats (above 80 gm.), of tetraploid nuclei. In the growth stage between 25 and 80 gm., there is a remarkable replacement of the diploid nuclei by the tetraploid ones. However, in the liver of adult rats weighing more than 150 gm., any increase or decrease in the frequency of diploid and tetraploid nuclei is hardly observable. In such rats, the nuclear population of the liver parenchyma seems to reach a cell-ecological equilibrium which is considered to be a stable one. 4. It is shown that such nuclear populations and the total number of nuclei in a liver are controlled by the growth state, and not by the age. 5. The decrease in the total number of diploid nuclei and the increase in tetraploid nuclei in the growing livers of rats weighing from 40 up to 130 gm. can both be explained by the hypothesis that the tetraploid nuclei originate from the interphase diploid nuclei without involving mitosis. This hypothesis implies that mitosis is confined to the reproduction of diploid cells alone. 6. It is suggested that, in general, the synthesis of DNA does not necessarily result in the formation of visible mitotic chromosomes. 7. Mitotic time and generation time of diploid nuclei and the percentage of the tetraploidization from diploid nuclei are calculated and discussed.     

QUANTITATIVE ASPECTS OF CUTANEOUS SENSORY ADAPTATION. I

1. Adaptation of tactile receptors in the skin of the frog to excitation by an intermittent jet of air is measured and correlated with certain properties of a series of notched discs used to interrupt the air stream. 2. Adaptation in fifteen cases is found to be described by either one of two empirical formulas, or t = -k log f + C, for nine preparations t = a f^-b, for six preparations where f is the per cent frequency at time t and -k and -b are constants defining the rate of adaptation.     

QUANTITATIVE ANALYSIS OF RESPONSES FROM LATERAL-LINE NERVES OF FISHES. II

1. The lateral-line nerves of trout as well as those of catfish are found to discharge impulses spontaneously at a high frequency. 2. The frequency of nerve impulse discharge is measured as a function of the number of participating receptor groups (lateral-line sense organs). A quantitative analysis is made of the contribution to the total response made by each group of sense organs. 3. An analysis of the variability of the response is presented which makes it possible to estimate quantitatively the longitudinal extent of damage to the neuromasts due to surgical manipulation. 4. A method is described for recording the response of a single nerve fiber in the lateral-line trunk. 5. The frequency of the spontaneous discharge from the lateral-line nerve trunk when plotted as a function of temperature according to the Arrhenius equation yields a temperature characteristic of approximately 5000 calories. 6. The variability of the frequency of response as a function of temperature indicates the existence of temperature thresholds for the spontaneous activity of the neuromasts. 7. A possible basis for the spontaneous activity is considered. It is pointed out that the lateral-line system may serve as a model of the Purkinje cells of the cerebellum.     

MEASUREMENT OF INFLUENZA VIRUS-ANTIBODY REACTION BY QUANTITATIVE ELECTRON MICROSCOPY

Measurement of the weight of individual virus particles from untreated and antibody-treated populations was made by quantitative electron microscopy. The weight of antibody bound depended on the concentration of antibody in solution. One population of viruses exposed to an antibody concentration which resulted in 95% inhibition of hemagglutination showed a mass increase of 55%, corresponding to an absolute increase of 9.0 x 10^-17 g in the median value. Another population, whose hemagglutination inhibition assay was 64%, showed a 39% increase in mass corresponding to an absolute median increase of 7.3 x 10^-17 g. The larger viruses in each population bound a greater absolute amount of antibody than did the smaller ones, but the latter bound relatively more antibody in proportion to their mass. No cross-reactivity was found between the antibody to influenza A/PR8 and the influenza strain B/LEE. Influenza A/PR8 controls exposed to nonspecific gamma-globulin displayed a significant weight loss, at least in part owing to loss from the core, as judged from the electron micrographs.     

甜茶素含量测定

<正> 甜茶(Rubus suavissimus S.Lee)含一种高甜度的甜茶素,为斯替维醇(Steviol)和葡萄糖结合而成的二萜甙。 甜叶菊中斯替维甙的含量测定方法有:气相色谱法、薄层——气谱联合测定法、薄层层离——分光光度法、薄层扫描测定法等。甜茶中,甙的含量测定尚无人进行过研究,本文参照宫崎等人的薄层层离——分光光度法进行了研究。即首先把甙加酸水解为异     

QUANTITATIVE TRITIUM AUTORADIOGRAPHY OF MAMMALIAN CHROMOSOMES : I. The Basic Method

A technique has been developed that allows repeated autoradiographs to be made of the isotope distribution in the chromosomes of a single cell. A series of 10 separate autoradiographs were made of a Chinese hamster diploid male metaphase cell which had been labeled with tritiated thymidine during the first 15 minutes of its DNA synthesis period in the previous interphase. Each autoradiograph had low grain densities above the chromosomes so that quantitation was feasible. The separate autoradiographs were photographically combined into a single composite in which grain images were converted to lines oriented at right angles to the chromosome axis. The line densities were then measured with a recording microdensitometer to yield graphs reflecting the isotope distribution along each chromosome. The area under each graph was directly proportional to the total number of grains counted above the corresponding chromosome in the 10 separate autoradiographs. The distribution of isotope along the chromosomes was different for each chromosome, and in some cases homologs also differed in their early labeling patterns.     

从姜黄中提取姜黄素的研究

本文利用75％乙醇浸提姜黄得到姜黄素粗提物,然后分别采用吸附分离法和沉淀法对粗提物进一步分离提纯得到纯度较高的姜黄素产品。在吸附分离法提取姜黄素实验中,考察了不同洗脱剂组成对洗脱率的影响,实验发现90％乙醇洗脱效果最好;在沉淀法实验中,考察了不同pH值条件下的姜黄素沉淀量,发现pH值为7．0时,姜黄素沉淀量最大。实验结果表明,沉淀法得到的姜黄素产品纯度比吸附分离法得到的高,但沉淀法产品收率比吸附分离法低,吸附分离法比沉淀法更适合工业应用。     

发酵液内氨甲酰妥布拉霉素的定量测定

报道了应用一种以测量图象面积为指标的Ｚｙ－３００Ａ多功能抑菌圈测量仪,能够精确、稳定、快速地测定发酵液经薄层层析生物显迹后单组分氨甲酰妥布拉霉素的抑菌斑面积,然后从绘制的标准曲线中直接读取氨甲酸妥布拉霉素在发酵液中的含量,从而排除了其他组分对测定的干扰。试验证明,用Ｚｙ－３００Ａ多功能抑菌圈测量仪测定抗生素薄层层析生物显迹的抑菌斑,具有线性宽、精密度高、重复性强、操作简单方便等优点．本方法的建立为抗生素的纯品检定和多组分抗生素发酵、分离和纯化提供了准确的定量数据．     

丝状真菌组织DNA的提取

用CTAB法直接从丝状真菌的新鲜菌丝中提取DNA,并将所提取DNA进行随机引物多态性扩增(RAPD),得到了较清晰的扩增图谱,证明该法为提取真菌DNA以用于分子生物学研究的一种有效方法。