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PubMed2004

Phases, periphases, and interphases equilibrium by molecular modeling. I. Mass equilibrium by the semianalytical stochastic perturbations method and application to a solution between (120) gypsum faces.

The SASP (semianalytical stochastic perturbations) method is an original mixed macro-nano-approach dedicated to the mass equilibrium of multispecies phases, periphases, and interphases. This general method, applied here to the reflexive relation C(k)<=>mu(k) between the concentrations C(k) and the chemical potentials mu(k) of k species within a fluid in equilibrium, leads to the distribution of the particles at the atomic scale. The macroaspects of the method, based on analytical Taylor's developments of chemical potentials, are intimately mixed with the nanoaspects of molecular mechanics computations on stochastically perturbed states. This numerical approach, directly linked to definitions, is universal by comparison with current approaches, DLVO Derjaguin-Landau-Verwey-Overbeek, grand canonical Monte Carlo, etc., without any restriction on the number of species, concentrations, or boundary conditions. The determination of the relation C(k)<=>mu(k) implies in fact two problems: a direct problem C(k)=>mu(k) and an inverse problem mu(k)=>C(k). Validation of the method is demonstrated in case studies A and B which treat, respectively, a direct problem and an inverse problem within a free saturated gypsum solution. The flexibility of the method is illustrated in case study C dealing with an inverse problem within a solution interphase, confined between two (120) gypsum faces, remaining in connection with a reference solution. This last inverse problem leads to the mass equilibrium of ions and water molecules within a 3 A thick gypsum interface. The major unexpected observation is the repulsion of SO(4) (2-) ions towards the reference solution and the attraction of Ca(2+) ions from the reference solution, the concentration being 50 times higher within the interphase as compared to the free solution. The SASP method is today the unique approach able to tackle the simulation of the number and distribution of ions plus water molecules in such extreme confined conditions. This result is of prime importance for all coupled chemical-mechanical problems dealing with interfaces, and more generally for a wide variety of applications such as phase changes, osmotic equilibrium, surface energy, etc., in complex chemical-physics situations.

Journal Article↗
PubMed1995

Evidence for the organization of chromatin in megabase pair-sized loops arranged along a random walk path in the human G0/G1 interphase nucleus.

We determined the folding of chromosomes in interphase nuclei by measuring the distance between points on the same chromosome. Over 25,000 measurements were made in G0/G1 nuclei between DNA sequences separated by 0.15-190 megabase pairs (Mbp) on three human chromosomes. The DNA sequences were specifically labeled by fluorescence in situ hybridization. The relationship between mean-square interphase distance and genomic separation has two linear phases, with a transition at approximately 2 Mbp. This biphasic relationship indicates the existence of two organizational levels at scales > 100 kbp. On one level, chromatin appears to be arranged in large loops several Mbp in size. Within each loop, chromatin is randomly folded. On the second level, specific loop-attachment sites are arranged to form a supple, backbonelike structure, which also shows characteristic random walk behavior. This random walk/giant loop model is the simplest model that fully describes the observed large-scale spatial relationships. Additional evidence for large loops comes from measurements among probes in Xq28, where interphase distance increases and then locally decreases with increasing genomic separation.

Cell Cycle↗
PubMed1967

Synthesis of RNA in mammalian cells during mitosis and interphase.

Chinese hamster cells in the mitotic and G(1) phases of the growth cycle were incubated for 30 or 60 min in suspension tissue culture and pulse-labeled with tritiated uridine. After appropriate chases, washes, and extractions, it was found that all incorporation into the nucleic acid may be accounted for by those cells in interphase. An average of 410 counts was found for incorporation into the cell population (approximately 2.0 x 10(5) cells) of which over 80% of the cells was initially in mitosis. The increasing number of cells leaving mitosis and entering interphase during the 30 min incubation was theoretically able to account for 470 counts. In addition, short-pulse labeling experiments have shown a consistent linear relationship between the percentage of cells in division and the incorporation of the isotope, which strongly suggests that, if 100% of the cells were in mitosis, the counts would be essentially zero. Thus, the entire label may be attributed to those cells in interphase where portions of the chromosomal material are known to be already extended.

Animals↗
PubMed1971

Induction of nuclear envelopes around metaphase chromosomes after fusion with interphase cells.

The process of cellular fusion induced by Sendai virus in Chinese hamster cells (Don line) afforded us the opportunity to study nuclear envelope formation around metaphase sets in the presence of interphase nuclei, when chromosome pulverization failed to occur in such multinucleate cells. Morphologically, the enveloped metaphase chromosomes resembled a normal telophase nucleus, though minor differences prompted us to call it telophase-like. Electron microscopic observations demonstrated that the membranes enveloping the chromosomes appeared to be identical with a normal nuclear envelope. The longer the cells were incubated with Colcemid before fusion, the higher was the number of cells with telophase-like nuclei and the lower the percentage of cells with pulverizations. Furthermore, the number of pulverizations bore a somewhat direct relationship to the ratio of metaphase to interphase nuclei in multinucleate cells, and the number of telophase-like nuclei was inversely proportional to this ratio. A hypothesis is advanced in which a balance between the activities of a chromosome pulverization factor and a nuclear envelope formation factor, the former in metaphase cells and the latter in interphase cells, is decisive as to the nature of morphologic events observed in virus-induced fused cells.

Animals↗
PubMed1973

Contrast between the environmental pH dependencies of prophasing and nuclear membrane formation in interphase-metaphase cells.

In Chinese hamster Don cells, fusion of an interphase cell with a metaphase cell resulted either in prophasing of the interphase nucleus, including loss of the nuclear envelope (NE), or in the formation of a double membrane around the metaphase chromosomes. Only one of these phenomena occurred in a given interphase-metaphase (I-M) binucleate cell. At pH 7.4, there was about an equal probability that either event could occur amongst the population of I-M cells. The effect of pH changes in the medium containing the fused cells was examined. At pH 6.6, prophasing was the predominant event; at pH 8.0, membrane formation predominated. It was found that the rate of progression of a mononucleate cell from G(2) to metaphase was appreciably faster at pH 6.6 than at pH 8.0. Conversely, the progression from metaphase to G(1) was faster at pH 8.0 than at pH 6.6. These results with the mononucleate cells strengthen the hypothesis that structural changes in I-M cells are reflections of normal mitotic phenomena. Additional evidence for this hypothesis was produced by electron microscope examination after direct fixation in chrom-osmium. The double membrane around the chromosomes of the I-M cell was indistinguishable from the normal NE. The results obtained by varying the pH of the medium containing the fused cells provide an indication that disruption or formation of the NE of Don cells depends on the balance reached between disruptive and formative processes.

Animals↗
PubMed1998

Pelizaeus-Merzbacher disease: identification of Xq22 proteolipid-protein duplications and characterization of breakpoints by interphase FISH.

Pelizaeus-Merzbacher disease (PMD) is an X-linked, dysmyelinating disorder of the CNS. Duplications of the proteolipid protein (PLP) gene have been found in a proportion of patients, suggesting that, in addition to coding-region or splice-site mutations, overdosage of the gene can cause PMD. We show that the duplication can be detected by interphase FISH, using a PLP probe in five patients and their four asymptomatic carrier mothers. The extent of the duplication was analyzed in each family by interphase FISH, with probes from a 1. 7-Mb region surrounding the PLP gene between markers DXS83 and DXS94. A large duplication >=500 kb was detected, with breakpoints that differed, between families, at the proximal end. Distinct separation of the duplicated PLP signals could be seen only on metaphase chromosomes in one family, providing further evidence that different duplication events are involved. Quantitative fluorescent multiplex PCR was used to confirm the duplication in patients, by the detection of increased copy number of the PLP gene. Multiallelic markers from the duplicated region were analyzed, since the identification of two alleles in an affected boy would indicate a duplication. The majority of boys were homozygous for all four markers, compared with their mothers, who were heterozygous for one to three of the markers. These results suggest that intrachromosomal rearrangements may be a common mechanism by which duplications arise in PMD. One boy was heterozygous for the PLP marker, indicating a duplication and suggesting that interchromosomal rearrangements of maternal origin also can be involved. Since duplications are a major cause of PMD, we propose that interphase FISH is a reliable method for diagnosis and identification of female carriers.

Central Nervous System Diseases↗
PubMed1979

The similarity of DNA sequences remaining bound to scaffold upon nuclease treatment of interphase nuclei and metaphase chromosomes.

The fragments of DNA attached to protein skeleton of interphase nuclei or metaphase chromosomes were obtained. Both the method involving restriction endonuclease treatment/1,2/and a novel procedure based on mild staphylococcal nuclease digestion were used. In the latter case, DNA fragments remaining bound to nuclei or chromosomes are not enriched in satellite but only in abundant middle repetitive DNA. The shorter the fragments of attached DNA, the higher the content of middle repetitive DNA in the fraction. It has a slightly higher density in a CsCl gradient comparing to the main DNA. The yield of attached DNA, its distribution in a CsCl density gradient, and its renaturation properties are essentially the same for interphase and metaphase chromosomes. The average size of DNA loops was found to be equal to approximately 60 kb for both metaphase chromosomes and interphase nuclei. The conclusion has been drawn that the bulk of attachment sites of DNP fibrils to axial chromosomal structures remains unchanged during the cell cycle.

Animals↗
PubMed2005

Identification of tumor entities of renal cell carcinoma using interphase fluorescence in situ hybridization.

PURPOSE: We developed a rapid interphase fluorescence in situ hybridization (FISH) test to differentiate renal cell carcinoma (RCC) based on known genetic alterations and verified the suitability of this test for practical use. MATERIALS AND METHODS: We composed 2 FISH test sets using 6 centromere specific and 2 region specific DNA probes of human chromosomes. Test set 1 contained centromeric probes for chromosomes 1, 2, 6 and 9, as labeled by 4 fluorescence dyes. For test set 2 we chose 3p24pter and 3p13p14 regions, and centromeric probes of chromosomes 7 and 17. Interphase nuclei of tumor specimens were prepared from 50 mum frozen tissue sections and fixed on slides. The 2 sets were hybridized simultaneously side by side on the same slide. RESULTS: Seven clear cell carcinomas, 8 papillary carcinomas, 7 chromophobe RCCs and 3 oncocytomas were analyzed by interphase FISH. Results were compared with comparative genomic hybridization findings and pathological reports. Genetic alterations were detected in 22 of 25 analyzed tumors by FISH. FISH findings absolutely correlated with comparative genomic hybridization results. Of the analyzed carcinomas 22 could be identified correctly. In 3 tumors the histological subtypes were revised. CONCLUSIONS: The results of this study demonstrate that the performed test set allows the accurate identification of RCC in 1 hybridization step. Therefore, FISH represents an effective method for the rapid classification of RCC.

Carcinoma, Renal Cell↗
PubMed1997

Probing the Plant Actin Cytoskeleton during Cytokinesis and Interphase by Profilin Microinjection.

We have examined the cytological effects of microinjecting recombinant birch profilin in dividing and interphase stamen hair cells of Tradescantia virginiana. Microinjection of profilin at anaphase and telophase led to a marked effect on cytokinesis; cell plate formation was often delayed, blocked, or completely inhibited. In addition, the initial appearance of the cell plate was wrinkled, thin, and sometimes fragmented. Injection of profilin at interphase caused a thinning or the collapse of cytoplasmic strands and a retardation or inhibition of cytoplasmic streaming in a dose-dependent manner. Confocal laser scanning microscopy of rhodamine-phalloidin staining in vivo revealed that high levels of microinjected profilin induced a degradation of the actin cytoskeleton in the phragmoplast, the perinuclear zone, and the cytoplasmic strands. However, some cortical actin filaments remained intact. The data demonstrate that profilin has the ability to act as a regulator of actin-dependent events and that centrally located actin filaments are more sensitive to microinjected profilin than are cortical actin filaments. These results add new evidence supporting the hypothesis that actin filaments play a crucial role in the formation of the cell plate and provide mechanical support for the cytoplasmic strands in interphase cells.

Journal Article↗
PubMed2001

Rapid detection of chromosome aneuploidies by prenatal interphase FISH (fluorescence in situ hybridization) and its clinical utility in Japan.

OBJECTIVE: The purpose of this study was to assess the accuracy, informative rate, detection rate, and clinical utility of prenatal interphase fluorescence in situ hybridization (FISH) analysis of amniotic fluid samples from Japanese women. METHODS: Amniotic fluid specimens from 2,639 Japanese women were received for prenatal interphase FISH and chromosome analysis. A questionnaire was designed to evaluate FISH clinical utility by collaboration sites. RESULTS: Based on 2,319 tested samples, the accuracy (100%), informative (94%), and detection (87.6%) rates were all high. The accuracy (100%), informative (90.2%), and detection (90.0%) rates were also remarkable in third-trimester pregnancies. We perceive significant advantages from this test regarding medical management and patient satisfaction. CONCLUSIONS: This novel report shows that in Japan prenatal interphase FISH testing is highly informative and accurate, not only in second-trimester pregnancies but also in third-trimester pregnancies. This test provides advantages to both physicians and patients, provided that its capabilities and limitations are understood.

Adult↗
PubMed2002

Gravity-induced anomalies in interphase spacing reported for binary eutectics.

It has been reasoned that desirable microstructural refinement in binary eutectics could result from freezing in reduced-gravity. It is recognized that the interphase spacing in a binary eutectic is controlled by solute transport and that, on Earth, buoyancy-driven convection may enhance this. Hence, it has been presumed that the interphase spacing ought to decrease when a eutectic alloy is frozen under conditions of much-reduced gravity, where such buoyancy effects would be largely absent. The result of such speculation has been that many workers have frozen various eutectics under reduced gravity and have reported that, although some eutectics became finer, others showed no change, and some even became coarser. This reported varied behavior will be reviewed in the light of long term studies by the author at Queen's University, including recent microgravity studies in which samples of two eutectic alloy systems, MnBi-Bi and MnSb-Sb, were frozen under very stable conditions and showed no change in interphase spacing.

Journal Article↗
PubMed2005

Frequency of chromosome 17 aneuploidy in primary and recurrent pterygium by interphase-fluorescence in situ hybridization.

AIM: To investigate chromosome 17 numerical aberrations by using fluorescence in situ hybridization (FISH) in pterygia and to find out whether there is any association between chromosome 17 aneuploidy and recurrent pterygia. METHODS: Pterygium tissue samples were taken from 21 patients by surgical excision. Eighteen of them had primary and 3 had recurrent pterygium. Peripheral whole blood interphase cells obtained from 11 healthy subjects were assigned as control group. The cells from pterygium tissue and peripheral blood were incubated with a hypotonic solution and fixed in order to obtain interphase nuclei. FISH analysis with chromosome-17-specific alpha-satellite DNA probe was performed on both the interphase nuclei of pterygium tissue (of patients) and peripheral whole blood cells of controls. RESULTS: The mean percentage of chromosome 17 aneuploidy was 4.71% for the pterygia group and 4.41% for the controls. No significant difference of chromosome 17 aneuploidy was observed between the patients and the controls. When the group of patients with recurrences was compared with the group without recurrences, there was a significant difference in the frequency of chromosome 17 aneuploidy (U = 17, p = 0.029). CONCLUSIONS: Chromosome 17 aneuploidy is probably not an important factor in the formation of pterygium, but it may be related to recurrence.

Adult↗
PubMed2006

Interphase M-FISH applications using commercial probes in prenatal and PGD diagnostics.

Early, rapid and reliable diagnosis is of first priority in prenatal medicine. The combination of specific sonographic markers (e.g. nuchal translucency) and biochemical parameters in maternal serum (e.g. free beta-human chorionic gonadotropin, pregnancy-associated plasma protein A), has already dramatically improved the sensitivity of non-invasive first trimester risk screening in pregnancy. In invasive prenatal diagnosis, in addition to well-established chorionic villi short-term culture, interphase multi-colour-fluorescence in situ hybridisation (M-FISH) on uncultured amnion cells has become a reliable tool for the rapid detection of fetal aneuploidies. Interphase M-FISH applications have enabled the diagnosis of selected chromosomal abnormalities in single cells and, therefore, have also become an important diagnostic tool for preimplantation diagnosis (PGD). The development of commercially available probe sets, in particular, has led to a broad use of interphase M-FISH in prenatal and PGD diagnosis.

Amniocentesis↗
PubMed2003

True chemical composition profiles at polymer interphases: recovery from measurements distorted by instrumental broadening.

The determination of chemical composition profiles at polymer interphases is an important issue at the moment of elucidating the physical mechanisms that operate in polymer diffusion processes and for calculating diffusion parameters. Several techniques are available to measure these profiles, the most common being forward recoil spectroscopy, Rutherford backscattering spectrometry, nuclear reaction analysis, confocal Raman microspectroscopy (CRM), and scanning infrared microscopy. However, all these techniques are affected by the limited resolution of the experimental setup, which in practice produces a rounding effect on the sharp corners of the composition profile; this may lead to incorrect conclusions regarding the measurements. In this work an inverse technique is proposed to correct this undesirable effect in the profiles. The inversion is performed on a model of the measuring process, which includes the instrumental broadening function, a quantitative representation of the limited resolution. The proposed methodology was tested using numerically generated experiments and genuine experimental runs obtained from CRM measurements at interphases of polymer bilayers. In all cases, the recovered profiles were close to the expected ones. In the truly experimental results diffusion tails are observed behind and ahead of the diffusion front before the numerical treatment of the data. These tails may be caused by a genuine mass diffusion or by an artifact. After the numerical treatment the tails disappear and a sharp interphase is recovered, a result one expects for the polymer pairs under study.

Computer Simulation↗
PubMed2003

Argyrophilic nucleolar organizer regions (AgNORs) in interphases and metaphases of normal and neoplastic gill cells of Macoma balthica (Bivalvia: Tellinidae) from the Gulf of Gdansk, Baltic Sea.

Chromosome analysis of gill cells of different populations of Macoma balthica (L.) from the Bay of Gdansk (Baltic Sea) revealed 2 clam categories, 1 with neoplastic features and 1 without. Silver-staining was performed on interphase and metaphase cells of both categories. The mean argyrophilic nucleolar organizer region (AgNOR) count per abnormal interphase cell was significantly higher than in normal interphase cells. Normal silver-stained metaphases had 3 nucleolar organizer region (NOR) chromosome phenotypes. The location of the NORs in the most frequent phenotype (55.6% in 54 metaphases scored) was interstitial on the largest metacentric chromosome pair, Pair No. 1. Abnormal silver-stained metaphases had a higher number of active NOR sites. Different phenotypes were observed (frequency greater than 10% for 67 metaphases scored); 2 were similar to those in normal metaphases and 5 were ectopic. The higher activity of AgNORs observed in abnormal cells confirmed the diagnosis of malignant neoplasia.

Animals↗
PubMedDate not supplied

[Rheological studies of interphase adsorption layers of lysozyme at phase separation liquid boundaries].

Rheological studies of thin layers of biopolymers provide a principally new approach to the studies of the structure and functions of biomembranes. Rheological properties of interphase adsorption layers of lysozyme at different concentrations, temperatures and at the border with various carbohydrates. The values of moduli of fast elastic deformation E1s, slow elastic deformation E2s, equilibrium module of elasticity Es, viscosity of elastic aftereffect, plastic viscosity eta 0s and eta s for the interphase layers of lysozyme at liquid borders show that the interphase adsorption layers were solidlike, and the moduli of dimeric structure evaluated as volume ones correspond to the characteristics of normal elastomeres.

Adsorption↗
PubMed2004

[Detection of abnormal numbers of chromosome 8 with interphase fluorescence in situ hybridization in hematologic malignancies].

OBJECTIVE: To explore the value of interphase fluorescence in situ hybridization (FISH) in the detection of abnormal numbers of chromosome 8 in patients with hematologic malignancies. METHODS: Conventional cytogenetics(CC) and interphase FISH using chromosome 8 centromere specific probe were simultaneously carried out to detect the abnormal numbers of chromosome 8 in eight acute myeloid leukemia cases with CC unveiled abnormal numbers of chromosome 8, ten chronic myeloid leukemia cases in accelerated phase or blast crisis, and three normal individuals. RESULTS: Nine cases that displayed trisomy 8 by means of CC were confirmed by FISH. Among them, Case 5 only displayed diploidy 8, trisomy 8 and tetrasomy 8 by CC, at the same time, FISH confirmed the presence of trisomy 8 and tetrasomy 8 and also revealed a low percentage of a pentasomy 8 clone. Case 3 and Case 17 had each only one cell with trisomy 8 by means of CC, and this could not determine whether they had the trisomy 8 clone, yet FISH confirmed the existence of trisomy 8 clone. Case 9 did not display trisomy 8 by CC, but FISH revealed the existence of trisomy 8 clone. In the other cases, the percentages of trisomy 8 cells determined by FISH were close to or significantly lower than those by CC, but for Case 16 where the percentage of trisomy 8 cells by FISH was significantly higher than that by CC. CONCLUSION: Interphase FISH is a useful method for the detection of abnormal numbers of chromosome 8, especially in the patients with normal or unsure karyotype or with less and bad metaphases. It is an important complement to CC.

Acute Disease↗
PubMed1976

[C- and Q-chromatin of the experimentally condensed human interphase chromosomes].

Condensed interphase chromosomes of the cultured human lymphocytes obtained by the fusion of interphase and metaphase cells were studied using C- and Q-bands techniques. The appearance and localization of the constitutive heterochromatin blocks on condensed chromosomes at G1-period were the same as on the metaphase ones. These characters were used for a group and individual identification of some chromosomes condensed at G1-period and for a study of the association of the constitutive heterochromatin blocks in the interphase nuclei. The fluorescent analysis of the chromosomes condensed at G1-period detected some bright fluorescent blocks of the constitutive heterochromatin.

Cell Division↗