Nuclear DNA,nuclear area and nuclear dry mass in thirteen species of Crotalaria (Angiospermae,Leguminosae)

Estimations on nuclear DNA content, nuclear area and nuclear dry mass were made on 13 species of Crotalaria, with the help of Vicker's M 85 microdensitometer and Vicker's interferometer. Highly significant differences for all three characters were found between species. DNA density (DNA/area) increased with increase in DNA content. The cultivated species, C. juncea had a lower DNA density relative to its DNA content.—Nuclear DNA made only a small fraction of nuclear dry mass (115). Although total dry mass and non-nucleolar nuclear dry mass showed significant regression on DNA content, the nucleolar dry mass gave a much wider scatter, indicating that nucleolar mass is not equally dependent on nuclear DNA content.     

The relationship of DNA synthesis to protein accumulation in the cell nucleus

The relationship between DNA synthesis and protein accumulation in cell nucleus and cytoplasm has been investigated by the use of a combination of ultramicrointerferometric and ultramicrospectrophotometric methods. 5-Fluoro-2'-deoxyuridine (FUdR) inhibited DNA synthesis, resulting in inhibition of cell proliferation in G-1 and early S-phase. However, synthesis and accumulation of protein continued in the presence of FUdR, as indicated by a 54% increase in the average dry mass value per individual cell during 18-hour exposure to FUdR; due primarily to protein accumulation in the cytoplasm, the average cytoplasmic dry mass increased by as much as 85%, while the dry mass of the nucleus increased by only 21%. The dry mass values of individual nuclei were well-correlated to the nuclear DNA content throughout the period of exposure to FUdR. In contrast to the continued accumulation of protein in the cytoplasm during inhibition of DNA synthesis, protein accumulation in the nucleus was inhibited. When cells were released from inhibition of DNA synthesis by the addition of 2'-deoxythymidine, the nuclear DNA content and nuclear dry mass increased in near-synchrony, there being some evidence that DNA synthesis was initiated somewhat prior to initiation of increase in nuclear dry mass. Thus, it appears that DNA synthesis (or an increase in nuclear DNA content) is intimately related to the regulation of protein accumulation in the nucleus.     

Heavy metal action on the dry mass content and surface area of nuclei and cytoplasm during differentiation of cortex cells in pea (Pisum sativum L.) roots

The cortex cells of pea roots (Pisum sativum L.) grown for 144 h in the presence of cadmium, chromium and lead at the concentration 10(-4) M were the object of the present studies. Applied metals reduced dry mass content and concentration of nuclei in meristematic and differentiation zones. Chromium only enhanced nuclear mass concentration in the differentiation zone. The metals also made dry mass content and concentration of cytoplasm reduce, but they diminished mostly the concentration of cytoplasm in the meristematic zone and its dry mass in differentiation zone. Stimulation of cytoplasmic dry mass concentration was visible in the 1st mm (Cr) and 7th mm (Cd). Moreover, chromium caused a marked increase of cytoplasmic dry mass content in the 3rd mm of root. The studied metals reduced nuclear size, calculated as surface area, in the meristematic and differentiation zones. The increment of nuclear dimensions was observed only in the 1-3rd mm (Pb), 3rd (Cr) and 7th mm (Cd). In the presence of the applied metals the surface area of cytoplasm increased only in the 3rd mm and in 5th mm (chromium only). The present observations have shown that the toxicity of studied metals is as follows; Cd greater than Pb greater than Cr (nucleus - dry mass content and concentration, cytoplasmic area), Pb greater than Cd greater than Cr (cytoplasmic dry mass content and concentration and Cd greater than Cr greater than Pb (nuclear dimensions).     

Calcium influence on the dry mass content and surface area of nuclei and cytoplasm during differentiation of cortex cells in pea (Pisum sativum L.) roots treated with heavy metals

The effect of Ca addition to Cd, Cr and Pb solutions on the nuclear and cytoplasmic dry mass content and its concentration as well as on these organelles dimensions were studied in cortex cells of pea roots. Ca alone, at the concentration 10(-8)M brought about a decrease (in comparison to water) in the dry mass content of nuclei and its concentration, but the increment was almost twice in the dry mass content of cytoplasm; however, it has no significant effect on its concentration. Ca ions addition does not change the surface area of nuclei except the 1st and 5th mm segments but causes a doubling of the area occupied by cytoplasm. In response to Ca addition to Cd or Cr solutions a further diminution of nuclear dry mass content takes place. Only in the case of Pb nuclear dry mass increases in the 7th mm or is similar in remaining root segments. The diminution of nuclear dry mass content due to Ca presence in metal solutions is accompanied by a lowering in its concentration, although in the presence of Cd and Pb the diminution is not significant. Ca ions addition results in an increase in cytoplasmic dry mass content. No such regularities were observed in the 1st (Cd, Pb) and 3rd (Cr) root segments. In response to Ca ions the concentration of cytoplasmic dry mass content increased insignificantly in differentiation zone and underwent reducation in the meristematic zone--in the 1st mm (Cr) and 3rd (Cd). After Ca addition to studied metal solutions the decrease in nuclear dimensions was visible only in Cd or Pb treated cells in the 3rd and 7th or in the 1st mm, respectively. An increase in nuclear size occurred only in Cr treated cells in the 7th mm. Enrichment of heavy metals with Ca caused the marked enlargement in cytoplasmic area in differentiation zone but the increment in it in meristematic zone was observed only in Cd (1st, 3rd) and Cr (1st) treated cells.     

Nuclear Dry Mass and Area in Epidermal Cells of Senescing Soybean Cotyledons

The nuclear dry mass and area of epidermal cells of senescingsoybean cotyledons were measured with an interference microscopeand found to decrease until cotyledon abscission. Decapitationafter germination prevented yellowing and abscission of cotyledonsand the decrease of both nuclear dry mass and area. The meansof nuclear dry mass and area for normal and decapitated cotyledonswere highly correlated in two experiments, r=0.0.76 and 0.90respectively.     

Mass and molecular weight of isolated nuclear rings

Nuclear rings are cell structures found at the nuclear cortex wedged between the nuclear envelope and the chromatin fiber network. In previous publications we have dealt with their morphology, relationships with the nuclear membranes, chromatin fibers and cytoskeletal filaments; and more recently, with their measurements at high electron microscope resolution. In this article we have calculated the mass and molecular weight of 336 isolated nuclear rings from human circulating lymphocytes using a photometric procedure and polystyrene latex spheres as the standard for weight calibration. Our results show a range of mass of 0.4-35.5 x 10(-16) g (equivalent to 0.2-21.2 x 10(8) Da with a positively skewed distribution (median: 3.3 x 10(-16) g or 2.0 x 10(8) Da). Mass and volume of nuclear rings were highly correlated. In addition, it was possible to calculate the area, the whole mass and the mass per unit area of the nuclear envelope present in the center of the nuclear rings. The mass of this area also shows a lognormal distribution (median of mass/unit area: 37.3 x 10(-8) pg/nm2 or 1.9 x 10(5) Da/nm2). We discuss the significance of this results as parameters for the characterization of the nuclear rings and their possible implications for a new interpretation of nuclear cortex architecture, nucleocytoplasmic traffic and macromolecule segregation between the two main cell compartments.     

Changes in the Nucleus during Cellular Development in the Pea Seedling

Nuclei were isolated from three regions of the root and fromthe epicotyls of growing pea seedlings. As the dry mass of thenuclei was the same before and after isolation, and as the DNAcontent of the nuclei accounted for the DNA of the cells, theisolated nuclei are assumed to be a random sample of the nucleiin the intact tissues. The DNA, RNA, and protein contents percell were much greater in the epicotyl than in the root, butthe RNA and protein contents of the nucleus were similar inboth epicotyl and root. Although the amounts of DNA, RNA, andprotein per cell increased with increasing distance from theroot tip, increase in the protein content of the nucleus occurredmainly between the meristematic and expanding regions, and atthis same point nuclear RNA decreased. Nuclear volume increasedwith increasing distance from the root tip, whereas nucleardry mass remained virtually unchanged. The nucleus thereforebecomes increasingly hydrated as the cells mature. The datasuggest a changing interaction between the nucleus and the cytoplasmduring cellular development.     

Nuclear protein redistribution in heat-shocked cells

An increase was observed in the total protein mass of nuclei isolated from Chinese hamster ovary cells heated at 45°C or 45.5°C. An increase in the fractional recovery of DNA polymerase α and β, and of DNA topoisomerase activity coincided with this increase in the protein mass of nuclei from heated cells. Nuclear protein mass which was soluble in 2.0 M NaCl decreased 0.5 fold, while DNA-associated and nuclear matrix-associated protein mass increased 2.2 and 3.4 fold, respectively. The results indicate that the increase in nuclear protein mass observed in nuclei from heated cells is due in part to an increased binding, or precipitation, of nuclear proteins onto the cell's DNA and nuclear matrix. © 1993 Wiley-Liss, Inc.     

Identification of a novel nuclear domain

For most known nuclear domains (ND), specific functions have been identified. In this report we used murine mAbs and human autoantibodies to investigate precisely circumscribed structures 0.2-0.3 micron in diameter which appear as "nuclear dots" distributed throughout the nucleoplasm. Nuclear dots are metabolically stable and resistant to nuclease digestion and salt extraction. The localization of nuclear dots is separate from kinetochores, centromeres, sites of mRNA processing and tRNA synthesis, nuclear bodies, and chromosomes. The nuclear dots, therefore, represent a novel ND. Nuclear dots break down as cells enter metaphase and reassemble at telophase. In interphase cells, nuclear dots are frequently "paired," and some are visible as "doublets" when stained with one particular antiserum. The number of dot doublets increased when quiescent cells were stimulated with serum although the total number of dots did not change substantially. One of the antigens was identified as a protein with a molecular mass of approximately 55 kD showing three charge isomers in the pI range of 7.4 to 7.7. Autoantibodies affinity purified from this nuclear dot protein (NDP-55) show nuclear dots exclusively. Nuclear dot-negative rat liver parenchymal cells became positive after chemical hepatectomy, suggesting involvement of the NDP-55 in the proliferative state of cells.     

Mass and molecular weight of isolated nuclear rings

Nuclear rings are cell structures found at the nuclear cortex wedged between the nuclear envelope and the chromatin fiber network. In previous publications we have dealt with their morphology, relationships with the nuclear membranes, chromatin fibers and cytoskeletal filaments; and more recently, with their measurements at high electron microscope resolution. In this article we have calculated the mass and molecular weight of 336 isolated nuclear rings from human circulating lymphocytes using a photometric procedure and polystyrene latex spheres as the standard for weight calibration. Our results show a range of mass of 0.4–35.5 × 10⁻¹⁶g (equivalent to 0.2–21.2 × 10⁸ Da with a positively skewed distribution (median: 3.3 × 10⁻¹⁶g or 2.0 × 10⁸ Da). Mass and volume of nuclear rings were highly correlated. In addition, it was possible to calculate the area, the whole mass and the mass per unit area of the nuclear envelope present in the center of the nuclear rings. The mass of this area also shows a lognormal distribution (median of mass/unit area: 37.3 × 10⁻⁸ pg/nm² or 1.9 × 10⁵ Da/nm²). We discuss the significance of this results as parameters for the characterization of the nuclear rings and their possible implications for a new interpretation of nuclear cortex architecture, nucleocytoplasmic traffic and macromolecule segregation between the two main cell compartments.     

Chromatin motion in neuronal interphase nuclei: changes induced by disruption of intermediate filaments

Motion of nucleoli within interphase nuclei, known as nuclear rotation, may be used as a measure of motion of chromatin domains within the global confines of the nucleus. Mechanisms by which chromatin domains are transposed remain enigmatic. It has been established that nuclei are anchored by a network of intermediate filaments, structural proteins which share epitopes with nuclear lamins and possibly representing a constraint on nuclear rotation. It is postulated that selective removal of this constraint, by acrylamide, would result in increased chromatin motion. Mean rates of nucleolar displacement were quantified in neurons, in vitro. Nuclear rotation increased from a mean control rate of 0.102 +/- 0.002 micron/min (n = 52) to a maximum mean rate of 0.207 +/- 0.026 micron/min (n = 11), after 23 hr of exposure to 4 mM acrylamide. Despite this significant increase in motion of intranuclear domains, cytoplasmic structures in the immediate juxtanuclear area did not exhibit increases in rates of motion. Immunocytochemistry was used to visualize cytoskeletal structures and to assay selective disruption of neurofilaments by acrylamide. Increased rates of chromatin motion coincided with breakdown of the intermediate filament network. Ultrastructural analyses showed that the increase in chromatin motion induced by acrylamide was also associated with a significant (P less than 0.005) change in the thickness of the nuclear lamina, decreasing from 20.9 +/- 5.10 nm (n = 159) in controls to 18.9 +/- 3.1 nm (n = 148), to 19.5 +/- 3.6 nm (n = 240) and to 16.1 +/- 4.4 nm (n = 103) at 4, 8 and 22 hr exposure, respectively. Moreover, the number of mitochondria per unit area changed significantly (P less than 0.0001) with exposure to acrylamide, increasing from 9.1 +/- 2.2 mitochondrial profiles in controls to 16.5 +/- 5.3 profiles after 22 hr exposure to acrylamide. Distribution of other cytoskeletal components, actin and microtubules, was not altered and does not appear to play a significant role in the observed increase in rates of nuclear rotation. We conclude that the removal of the damping effects on chromatin motion normally imposed by the nuclear lamina and by intermediate filaments results in increased chromatin motion.     

Temporal effects of progesterone domination on estrogen and oxytocin receptors in hamster uterus

The purpose of this study was to determine whether progesterone (P)-induced down regulation of estrogen receptors (Re) and oxytocin receptors (ROT) changes with the time of P exposure. Ovariectomized hamsters were given s.c. Silastic implants of estradiol (E2) and P for 4, 8 and 16 days. Cytosol and nuclear Re were measured at low temperature with the pyridoxal phosphate exchange assay, and ROT was assayed in the membrane fraction by [3H] oxytocin binding. Nuclear Re and ROT were down regulated throughout the 16-day P exposure period, but cytosol Re (and total Re) increased progressively from 4 to 16 days indicating that the down regulation of cytosol Re escapes P control with time. This conclusion was supported by P withdrawal studies in which P implants were removed for 6 or 12 h. P withdrawal resulted in equivalent recovery responses of nuclear Re and ROT after 4, 8 and 16 days of P exposure. Although cytosol Re recovery to P withdrawal occurred at 4 and 8 days, no response was obtained after 16 days of P exposure. Uterine weight increased during steroid treatment, and morphometric analysis of the P-dominated uterus revealed significant increases in the cross sectional area of the endometrium and myometrium with time of P exposure. Cytological examination of the uterus showed prominent secretory changes in the epithelial compartment on day 16 with accumulation of secretion in the uterine lumen. These results demonstrate that P can chronically down regulate nuclear Re and ROT. However, the control of cytosol Re varies with the time of P exposure, and cytosol Re levels become refractory to P domination by 16 days. The present observations indicate that the escape of cytosol Re from P control may be associated with the proliferation of one of more uterine cell populations such as glandular and luminal epithelial cells.     

Growth,abscisic acid content,and carbon isotope composition in wheat cultivars grown under different soil moisture

Changes in dry matter accumulation and allocation, abscisic acid content and carbon isotope composition of three wheat cultivars from dry, middle and wet climate regions were recorded at full maturity after exposure to different watering regimes (100, 50 and 25 % field capacity). Compared with the wet climate cultivar, the dry climate cultivar showed lower stem height, total leaf area, total dry biomass and total grain dry mass, and higher root/shoot ratio, abscisic acid content and carbon isotope composition under all watering regimes. Both water-limited treatments significantly reduced leaf growth and increased dry matter allocation into the roots leading to a significant raise of root/shoot ratio in all cultivars tested. In addition, drought affected morphological and physiological properties more in the dry climate cultivar than in the wet climate cultivar.     

Influence of long-term continuous illumination on the nuclear volume of hypothalamic neurons in female rats

A light microscopic quantitative analysis was performed for studying the effects of long-term exposure to continuous illumination on hypothalamic morphology in female rats. Nuclear volumes of neurons in the suprachiasmatic nucleus, paraventricular nucleus, ventromedial nucleus and preoptic area were measured in 16-18 month-old Long-Evans female rats which had been exposed to continuous light for 11 months. Significant nuclear volume shrinkage could be observed in all investigated regions except for the preoptic area in these animals when compared to that of control rats kept under a photoperiod of 14h light and 10h dark. The relationship between morphological changes of hypothalamic neurons and the marked alteration of melatonin and estrogen levels induced by long-term exposure to continuous light is discussed.     

Alterations in the nuclear matrix protein mass correlate with heat-induced inhibition of DNA single-strand-break repair

The total protein mass co-isolating with the nuclear matrix or nucleoid from Chinese hamster ovary (CHO) cells was observed to increase in heated cells as a function of increasing exposure temperature between 43 degrees C and 45 degrees C or of exposure time at any temperature. The sedimentation distance of the CHO cell nucleoid in sucrose gradients increased with increasing exposure time at 45 degrees C. Both these nuclear alterations correlated in a log-linear manner with heat-induced inhibition of DNA strand break repair. A two-fold threshold increase in nuclear matrix protein mass preceded any substantial inhibition of repair of DNA single-strand breaks. When preheated cells (45 degrees C for 15 min) were incubated at 37 degrees C the nuclear matrix protein mass and nucleoid sedimentation recovered with a half-time of about 5 h, while DNA single-strand-break repair recovered with a half-time of about 2 h. When preheated cells were placed at 41 degrees C (step-down heating; SDH) a further increase was observed in the nuclear matrix protein mass and the half-time of DNA strand break repair, while nucleoid sedimentation recovered toward control values. These results implicate alterations in the protein mass of the nuclear matrix in heat-induced inhibition of repair of DNA single-strand breaks.     

Noninvasive papillary transitional-cell tumors. Karyometric and DNA-content analysis

An investigation was undertaken to quantify and correlate the nuclear and DNA-content modifications in the different grades of noninvasive urothelial papillary carcinoma. Nuclear area and a roundness factor were studied by histomorphometry in 35 cases, and the DNA content was analyzed in 27 cases. The values of the nuclear area and the roundness factor increased from normal-looking urothelium to grade 3 tumors. The DNA-content analysis indicated a progressive change of ploidy and the presence of an aneuploid (greater than 4c) population in some of the grade 2 and grade 3 lesions. A high coefficient of correlation between the nuclear abnormalities and the DNA content was found.     

核辐射对大深度快速上浮脱险致减压病大鼠相关指标的影响*

目的: 探讨不同剂量核暴露后不同时间对大深度快速上浮脱险致减压病大鼠模型的发病率、死亡率及损伤指标的影响。方法: 80只SD雄性大鼠,随机分成空白对照组、脱险对照组和6个干预组(4 Gy辐射后4 h脱险、6 Gy辐射后4 h脱险、12 Gy辐射后4 h脱险、4 Gy辐射后8 h脱险、6 Gy辐射后8 h脱险、12 Gy辐射8 h后脱险),每组10只。干预组动物先采用不同剂量γ射线外照射(4、6、12 Gy),再进行大深度快速上浮脱险实验(最大加压深度150 m),分析大鼠肺W/D、脾指数及血浆IL-1β的变化。结果: 与脱险对照组比较,核辐射后脱险大鼠的减压病发病率及死亡率明显上升。4 Gy、6 Gy照射4 h后上浮脱险的大鼠发病率和死亡率较照射8 h后高。12 Gy辐射后4 h及8 h脱险大鼠的减压病的发病率及死亡率均比低剂量照射组明显增高,死亡率尤其明显。和发病率及死亡率的变化相一致,肺组织湿/干比、肺组织病理损伤程度、脾指数下降也表现同样的变化趋势:较低剂量(4 Gy、6 Gy)辐射后4 h改变明显,8 h改变不明显,而高剂量(12 Gy)辐射后4、8 h均变化明显。和空白对照组及脱险对照组相比较,各辐射后脱险组的血浆IL-1β浓度均显著上升。结论: 核辐射引起放射性肺损伤、免疫功能下降及血浆炎症因子浓度升高,会增加大鼠快速上浮脱险致减压病的风险。     

Nuclear changes in allium cepa in response to infection by fusarium moniliforme

Fusarium moniliforme was isolated [into culture] from white onion and inoculated into holes bored in onion bulbs. After 48 hrs, the nuclear dry mass of onion cells 0–5 mm ahead of the mycelium had decreased by 19% as determined by quantitative interference microscopy as compared to nuclear dry mass decreases of 11 % caused by avirulent Aspergillus niger, 26 % by an unidentified onion pathogen, 41 % by virulent Botrytis allii, and 42 % by virulent Aspergillus niger. In the present study, the nucleolar dry mass declined by 34 % in response to F. moniliforme under the same conditions. Degradation or exit of nuclear and nucleolar macromolecules in response to fungal products secreted in advance of mycelium may be responsible for lower nuclear and nucleolar dry mass.     

Carbon Accumulation during Photosynthesis in Leaves of Nitrogen- and Phosphorus-Stressed Cotton

Leaves of cotton (Gossypium hirsutum L.) accumulate considerable dry mass per unit area during photosynthesis. The percentage of C in that accumulated dry mass was estimated as the regression coefficient (slope) of a linear regression relating C per unit area to total dry mass per unit area. Plants were grown on full nutrients or on N- or P-deficient nutrient solutions. In the fully nourished controls, the mass that accumulated over a 9-hour interval beginning at dawn contained 38.6% C. N and P stress increased the C concentration of accumulated mass to 49.7% and 45.1%, respectively. Nutrient stress also increased the starch concentration of accumulated mass, but starch alone could not account for the differences in C concentration. P stress decreased the estimated rate of C export from source leaves, calculated as the difference between C assimilation and C accumulation. The effect of P stress on apparent export was very sensitive to the C concentration used in the calculation, and would not have been revealed with an assumption of unchanged C concentration in the accumulated mass.     

After Fukushima: managing the consequences of a radiological release

Even amidst the devastation following the earthquake and tsunami in Japan that killed more than 20,000 people, it was the accident at the Fukushima Daiichi nuclear power plant that led the country's prime minister, Naoto Kan, to fear for "the very existence of the Japanese nation." While accidents that result in mass radiological releases have been rare throughout the operating histories of existing nuclear power plants, the growing number of plants worldwide increases the likelihood that such releases will occur again in the future. Nuclear power is an important source of energy in the U.S. and will be for the foreseeable future. Accidents far smaller in scale than the one in Fukushima could have major societal consequences. Given the extensive, ongoing Nuclear Regulatory Commission (NRC) and industry assessment of nuclear power plant safety and preparedness issues, the Center for Biosecurity of UPMC focused on offsite policies and plans intended to reduce radiation exposure to the public in the aftermath of an accident. This report provides an assessment of Japan's efforts at nuclear consequence management; identifies concerns with current U.S. policies and practices for "outside the fence" management of such an event in the U.S.; and makes recommendations for steps that can be taken to strengthen U.S. government, industry, and community response to large-scale accidents at nuclear power plants.