scholarly journals Nuclear Position and Shape Deformation of Chromosome 8 Territories in Pancreatic Ductal Adenocarcinoma

2011 ◽  
Vol 34 (1-2) ◽  
pp. 21-33 ◽  
Author(s):  
Sylvia Timme ◽  
Eberhard Schmitt ◽  
Stefan Stein ◽  
Jutta Schwarz-Finsterle ◽  
Jenny Wagner ◽  
...  
Keyword(s):  
Nuclear Periphery ◽  
Nuclear Architecture ◽  
Neoplastic Cell ◽  
Carcinoma Cells ◽  
Chromosome 8 ◽  
Ductal Adenocarcinoma ◽  
Cell Nuclei ◽  
Formalin Fixed Paraffin ◽  
Transcriptional Alterations ◽  
Functional Relevance

Cell type specific radial positioning of chromosome territories (CTs) and their sub-domains in the interphase seem to have functional relevance in non-neoplastic human nuclei, while much less is known about nuclear architecture in carcinoma cells and its development during tumor progression. We analyzed the 3D-architecture of the chromosome 8 territory (CT8) in carcinoma and corresponding non-neoplastic ductal pancreatic epithelium. Fluorescence-in-situ-hybridization (FISH) with whole chromosome painting (WCP) probes on sections from formalin-fixed, paraffin wax-embedded tissues from six patients with ductal adenocarcinoma of the pancreas was used. Radial positions and shape parameters of CT8 were analyzed by 3D-microscopy. None of the parameters showed significant inter-individual changes. CT8 was localized in the nuclear periphery in carcinoma cells and normal ductal epithelial cells. Normalized volume and surface of CT8 did not differ significantly. In contrast, the normalized roundness was significantly lower in carcinoma cells, implying an elongation of neoplastic cell nuclei. Unexpectedly, radial positioning of CT8, a dominant parameter of nuclear architecture, did not change significantly when comparing neoplastic with non-neoplastic cells. A significant deformation of CT8, however, accompanies nuclear atypia of carcinoma cells. This decreased roundness of CTs may reflect the genomic and transcriptional alterations in carcinoma.

Chromosome condensation induced by geminivirus infection of mature plant cells

2000 ◽  
Vol 113 (7) ◽  
pp. 1149-1160 ◽  
Author(s):  
H.W. Bass ◽  
S. Nagar ◽  
L. Hanley-Bowdoin ◽  
D. Robertson
Keyword(s):  
Coat Protein ◽  
Chromatin Condensation ◽  
Nuclear Periphery ◽  
Nuclear Architecture ◽  
Plant Cells ◽  
Image Data ◽  
Chromosome Condensation ◽  
Fish Analysis ◽  
Viral Dna ◽  
Plant Dna

Tomato golden mosaic virus (TGMV) is a geminivirus that replicates its single-stranded DNA genome through double-stranded DNA intermediates in nuclei of differentiated plant cells using host replication machinery. We analyzed the distribution of viral and plant DNA in nuclei of infected leaves using fluorescence in situ hybridization (FISH). TGMV-infected nuclei showed up to a sixfold increase in total volume and displayed a variety of viral DNA accumulation patterns. The most striking viral DNA patterns were bright, discrete intranuclear compartments, but diffuse nuclear localization was also observed. Quantitative and spatial measurements of high resolution 3-dimensional image data revealed that these compartments accounted for 1–18% of the total nuclear volume or 2–45% of the total nuclear FISH signals. In contrast, plant DNA was concentrated around the nuclear periphery. In a significant number of nuclei, the peripheral chromatin was organized as condensed prophase-like fibers. A combination of FISH analysis and indirect immunofluorescence with viral coat protein antibodies revealed that TGMV virions are associated with the viral DNA compartments. However, the coat protein antibodies failed to cross react with some large viral DNA inclusions, suggesting that encapsidation may occur after significant viral DNA accumulation. Infection by a TGMV mutant with a defective coat protein open reading frame resulted in fewer and smaller viral DNA-containing compartments. Nevertheless, nuclei infected with the mutant virus increased in size and in some cases showed chromosome condensation. Together, these results established that geminivirus infection alters nuclear architecture and can induce plant chromatin condensation characteristic of cells arrested in early mitosis.

scholarly journals Effects of acetylsalicylic acid and acetic acid solutions in VX2 carcinoma cells: In vitro analysis

2006 ◽  
Vol 21 (3) ◽  
pp. 151-154 ◽  
Author(s):  
Rogério Saad-Hossne ◽  
René Gamberini Prado ◽  
William Saad Hossne
Keyword(s):  
Cell Death ◽  
Acetic Acid ◽  
Cell Viability ◽  
Acetylsalicylic Acid ◽  
Neoplastic Cell ◽  
Carcinoma Cells ◽  
Acid Solutions ◽  
Vx2 Carcinoma ◽  
Vitro Analysis

PURPOSE: To analyze, in vitro, the effects of acetylsalicylic acid (aspirin) and acetic acid solutions on VX2 carcinoma cells in suspension and to examine the correlation between these effects and neoplastic cell death. METHODS: The VX2 tumor cells (10(7) cells/ml) were incubated in solutions containing differing concentrations (2.5% and 5%) of either acetylsalicylic acid or acetic acid, or in saline solution (controls). Every five minutes, cell viability was tested (using the trypan blue test) and analyzed under light microscopy. RESULTS: Tumor cell viability (in %) decreased progressively and, by 30 minutes, neoplastic cell death had occurred in all solutions. CONCLUSION: Based on this experimental model and the methodology employed, we conclude that these solutions cause neoplastic cell death in vitro.

scholarly journals Automatic Quantification of Immunohistochemically Stained Cell Nuclei Based on Standard Reference Cells

1998 ◽  
Vol 17 (2) ◽  
pp. 111-123 ◽  
Author(s):  
Petter Ranefall ◽  
Kenneth Wester ◽  
Ann-Catrin Andersson ◽  
Christer Busch ◽  
Ewert Bengtsson
Keyword(s):  
Cyclin A ◽  
Cell Nuclei ◽  
Cultured Fibroblasts ◽  
Automatic Quantification ◽  
Formalin Fixed Paraffin ◽  
Ki67 Antigen ◽  
Formalin Fixed Paraffin Embedded ◽  
Fully Automatic ◽  
Formalin Fixed ◽  
Nuclear Texture

A fully automatic method for quantification of images of immunohistochemically stained cell nuclei by computing area proportions, is presented. Agarose embedded cultured fibroblasts were fixed, paraffin embedded and sectioned at 4 µm. They were then stained together with 4 µm sections of the test specimen obtained from bladder cancer material.A colour based classifier is automatically computed from the control cells. The method was tested on formalin fixed paraffin embedded tissue section material, stained with monoclonal antibodies against the Ki67 antigen and cyclin A protein. Ki67 staining results in a detailed nuclear texture with pronounced nucleoli and cyclin A staining is obtained in a more homogeneously distributed pattern.However, different staining patterns did not seem to influence labelling index quantification, and the sensitivity to variations in light conditions and choice of areas within the control population was low. Thus, the technique represents a robust and reproducible quantification method.In tests measuring proportions of stained area an average standard deviation of about 1.5% for the same field was achieved when classified with classifiers created from different control samples.

scholarly journals Nucleosome–nucleosome interactions via histone tails and linker DNA regulate nuclear rigidity

2017 ◽  
Vol 28 (11) ◽  
pp. 1580-1589 ◽  
Author(s):  
Yuta Shimamoto ◽  
Sachiko Tamura ◽  
Hiroshi Masumoto ◽  
Kazuhiro Maeshima
Keyword(s):  
Molecular Mechanisms ◽  
Mechanical Response ◽  
Nuclear Architecture ◽  
Living Cells ◽  
Genome Integrity ◽  
Biological Processes ◽  
Cell Nuclei ◽  
Histone Tails ◽  
Insight Into

Cells, as well as the nuclei inside them, experience significant mechanical stress in diverse biological processes, including contraction, migration, and adhesion. The structural stability of nuclei must therefore be maintained in order to protect genome integrity. Despite extensive knowledge on nuclear architecture and components, however, the underlying physical and molecular mechanisms remain largely unknown. We address this by subjecting isolated human cell nuclei to microneedle-based quantitative micromanipulation with a series of biochemical perturbations of the chromatin. We find that the mechanical rigidity of nuclei depends on the continuity of the nucleosomal fiber and interactions between nucleosomes. Disrupting these chromatin features by varying cation concentration, acetylating histone tails, or digesting linker DNA results in loss of nuclear rigidity. In contrast, the levels of key chromatin assembly factors, including cohesin, condensin II, and CTCF, and a major nuclear envelope protein, lamin, are unaffected. Together with in situ evidence using living cells and a simple mechanical model, our findings reveal a chromatin-based regulation of the nuclear mechanical response and provide insight into the significance of local and global chromatin structures, such as those associated with interdigitated or melted nucleosomal fibers.

scholarly journals Potentially Bioaccessible Phenolics from Mung Bean and Adzuki Bean Sprouts Enriched with Probiotic—Antioxidant Properties and Effect on the Motility and Survival of AGS Human Gastric Carcinoma Cells

Molecules ◽  
2020 ◽  
Vol 25 (13) ◽  
pp. 2963 ◽  
Author(s):  
Michał Świeca ◽  
Anna Herok ◽  
Katarzyna Piwowarczyk ◽  
Małgorzata Sikora ◽  
Patryk Ostanek ◽  
...  
Keyword(s):  
Antioxidant Properties ◽  
Human Stomach ◽  
Carcinoma Cells ◽  
Antioxidant Potential ◽  
Adzuki Bean ◽  
Cell Nuclei ◽  
Low Concentrations ◽  
Human Stomach Cancer ◽  
Human Gastric Carcinoma Cells ◽  
Bean Sprouts

Gastric digests from mung (MBS) and adzuki (ABS) bean sprouts enriched with probiotic Lactobacillus plantarum 299v were tested for their antioxidant potential, as well as antiproliferative and antimotility properties, in human stomach cancer cells (AGS). The digest of ABS contained quercetin and kaempferol derivates, while kaempferol and apigenin derivates were dominant in MBS. Compared to the controls, the probiotic-rich sprouts had a higher antioxidant potential—by 13% and 9%, respectively. Adzuki bean sprouts decreased the viability of AGS already at low concentrations (25% motility inhibitions). MBS and ABS displayed dose-independent cytostatic effects. The ABS extracts decreased the proliferation of AGS more effectively than the MBS extracts—0.2‰ ABS exerted c.a. 70% of inhibitions. Moreover, the phytochemicals from the probiotic-rich sprouts considerably reduced this activity. The increased vinculin level, the apoptotic shape of cell nuclei, and the reduced cell motility and proliferation indicate that the extracts exhibited cytostatic and cytotoxic activity.

Proteome of formalin-fixed paraffin-embedded pancreatic ductal adenocarcinoma and lymph node metastases

2012 ◽  
Vol 226 (5) ◽  
pp. 756-763 ◽  
Author(s):  
Kalnisha Naidoo ◽  
Richard Jones ◽  
Branko Dmitrovic ◽  
Nilukshi Wijesuriya ◽  
Hemant Kocher ◽  
...  
Keyword(s):  
Lymph Node ◽  
Pancreatic Ductal Adenocarcinoma ◽  
Lymph Node Metastases ◽  
Ductal Adenocarcinoma ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Node Metastases ◽  
Formalin Fixed

Spatial Distribution of Heterochromatin Bodies in the Nuclei of Triatoma infestans (Klug)

2020 ◽  
Vol 26 (3) ◽  
pp. 567-574
Author(s):  
Carlos Henrique L. Imperador ◽  
Vanessa B. Bardella ◽  
Eli Heber M. dos Anjos ◽  
Vera L.C.C. Rodrigues ◽  
Diogo C. Cabral-de-Mello ◽  
...  
Keyword(s):  
Spatial Distribution ◽  
18S Rdna ◽  
Constitutive Heterochromatin ◽  
Nuclear Periphery ◽  
Malpighian Tubule ◽  
Triatoma Infestans ◽  
Nuclear Region ◽  
Cell Nuclei ◽  
Variable Distance ◽  
Active Genes

AbstractConstitutive heterochromatin typically exhibits low gene density and is commonly found adjacent or close to the nuclear periphery, in contrast to transcriptionally active genes concentrated in the innermost nuclear region. In Triatoma infestans cells, conspicuous constitutive heterochromatin forms deeply stained structures named chromocenters. However, to the best of our knowledge, no information exists regarding whether these chromocenters acquire a precise topology in the cell nuclei or whether their 18S rDNA, which is important for ribosome function, faces the nuclear center preferentially. In this work, the spatial distribution of fluorescent Feulgen-stained chromocenters and the distribution of their 18S rDNA was analyzed in Malpighian tubule cells of T. infestans using confocal microscopy. The chromocenters were shown to be spatially positioned relatively close to the nuclear periphery, though not adjacent to it. The variable distance between the chromocenters and the nuclear periphery suggests mobility of these bodies within the cell nuclei. The distribution of 18S rDNA at the edge of the chromocenters was not found to face the nuclear interior exclusively. Because the genome regions containing 18S rDNA in the chromocenters also face the nuclear periphery, the proximity of the chromocenters to this nuclear region is not assumed to be associated with overall gene silencing.

Nuclear lamina and regulation of nuclear envelofe structure

1987 ◽  
Vol 45 ◽  
pp. 806-807
Author(s):  
Larry Gerace ◽  
Ueli Aebi ◽  
Brian Burke ◽  
Frank Suprynowicz
Keyword(s):  
Electron Microscopy ◽  
Nuclear Envelope ◽  
Xenopus Oocytes ◽  
Nuclear Periphery ◽  
Nuclear Architecture ◽  
Supramolecular Assembly ◽  
Nuclear Lamina ◽  
Eukaryotic Cells ◽  
Functional Studies ◽  
Tetragonal Lattice

The nuclear lamina is a protein meshwork that lines the nucleoplasmic surface of the nuclear envelope. In numerous higher eukaryotic cells, the lamina is known to contain a polymer of 1-3 major polypeptides (“lamins“) that form an insoluble supramolecular assembly during interphase. The lamina is thought to provide both a skeletal framework for the nuclear envelope (that regulates its disassembly and reformation during mitosis) and an anchoring site at the nuclear periphery for interphase chromosomes. Recent structural and functional studies on the nuclear lamina have yielded important new insight on its roles in nuclear architecture.Using electron microscopy, we found that the nuclear lamina of Xenopus oocytes is a meshwork of intermediate-sized (8-12 nm) filaments arranged in a near-tetragonal lattice having a spacing of 52 nm.

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