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At least 217 records · Page 12Linked to original sources
PubMed1994

Estimates of aneuploidy using multicolor fluorescence in situ hybridization on human sperm.

Single color fluorescence in situ hybridization (FISH) has been utilized on sperm to estimate nondisjunction rates for chromosomes 1, 12, 15, 16, X and Y. Using single-color FISH, one cannot distinguish nonhybridization from nullisomy nor disomy from diploidy. In order to provide an internal control, a multicolor FISH strategy was employed. Satellite probes specific for 13 human chromosomes were used on multiple semen samples from two normal donors. Two or three probes were hybridized simultaneously and scored by two independent observers. Over all experiments, 40,641 sperm were analyzed. The majority of autosomes had no significant difference in aneuploidy between chromosomes or between donors. However, a significant difference was observed for chromosome 18 between donors (chi 2(2) = 7.078, 0.025 < P < 0.05). Additionally, no significant difference was found between donors for sex chromosome aneuploidy. The frequency of sex chromosome aneuploidy was similar to that seen in paternally derived 47,XXY and 47,XYY conceptuses. Furthermore, 0.15% of sperm were found to be diploid. Based on the results of this study, as much as 19% of all sperm may be chromosomally abnormal. This method proved to be useful for determining aneuploidy of human chromosomes in sperm and valuable in exploring whether individual differences of nondisjunction exist.

Adult↗
PubMed1995

Multicolor FISH with a telomere repeat and Sry sequences shows that Sxr (Sex reversal) in the mouse is a new type of chromosome rearrangement.

XYSxr (Sex reversal) mice carry a Y chromosome in which the chromatin (including Sry, the gene for testis determination) that normally resides on the short arm is duplicated and the second copy is relocated to the distal end of the long arm. Multicolor in situ hybridization to mitotic chromosomes of XYSxr males using probes for the telomere repeat sequence (TTAGGG)n and Sry shows that the rearranged chromatin is located distal to the telomeric signal. This suggests that the rearrangement arose from a recombination event involving the distal Y telomere sequences, i.e., within the telomere, a structure historically assumed to be incapable of participating in chromosome rearrangements.

Animals↗
PubMed1996

A novel multicolor hybridization scheme applied to localization of a transcribed sequence (D10S170/H4) and deletion mapping in the thyroid cancer cell line TPC-1.

The sequence-tagged site (STS) D10S170, also referred to as H4, is a gene of unknown function. Its 5' end was found fused to the catalytic domain of the RET protooncogene to generate RET/PTC 1, the most common form of PTC oncogenes in human papillary thyroid carcinoma. This gene has previously been assigned to a very large genomic region, 10q11.22-->q22.1. Here, we describe the application of a novel hybridization scheme to the physical and genetic mapping of D10S170. First, we selected a homologous large-insert DNA clone from a human P1 library by filter hybridization and confirmed its authenticity by Southern blot analysis. Triple-color fluorescence in situ hybridization (FISH) experiments mapped this clone to l0q21.2-->q21.3. "Binning" experiments were performed using a quadruple-color FISH approach aimed toward placing the gene in a genetic interval defined by differentially labeled P1 DNA probes containing known polymorphic markers. We found that multicolor FISH greatly expedites chromosomal mapping. Finally, we applied our FISH approach to determine the extent of deletion involving this locus (D10S170) in a papillary thyroid cancer cell line, TPC-1.

Carcinoma, Papillary↗
PubMed2005

The detection of isolated tumor cells in bone marrow comparing bright-field immunocytochemistry and multicolor immunofluorescence.

BACKGROUND: The detection of isolated tumor cells in bone marrow by immunocytochemistry (ICC) has been reported to predict progression of early-stage breast cancer. The most common staining procedure uses bright-field ICC with cytokeratin (CK) antibodies to label isolated tumor cells. However, this method can result in false-positive staining events. We used multicolor immunofluorescence (IF) to develop a more specific assay for detecting isolated tumor cells in marrow samples from breast cancer patients. METHODS: We compared ICC and IF side by side for detection of cancer cells and false-positive staining events on bone marrow aspirates from breast cancer patients, bone marrow from healthy donors, and healthy donor blood spiked with cancer cells. The primary target for isolated tumor cell detection was CK for both methods. IF used an additional set of antibodies to label hematopoietic cells (HCs). RESULTS: The detection rate of CK+ events in breast cancer patient bone marrow aspirates was 18 (58%) of 31 for ICC and 21 (68%) of 31 for IF. However, with IF, 17 of 21 CK+ cases were stained with HC markers and thus were identified as false-positive events. A surprisingly high CK+ event rate was observed in healthy donor blood and marrow. In all healthy donor samples, CK+ events were readily identified as HCs by IF. Detection sensitivity of spiked cancer cells in donor blood was similar for both methods. CONCLUSIONS: There is a high frequency of CK+ events in blood and marrow, and it is important to note that this is observed both in patients with and those without cancer. IF with multiple HC markers allows straightforward discrimination between CK+ cells of hematopoietic and nonhematopoietic origin.

Bone Marrow Examination↗
PubMed2006

Isolation and characterization of a beta-primeverosidase-like enzyme from Penicillium multicolor.

p-Nitrophenyl and eugenyl beta-primeveroside (6-O-beta-D-xylopyranosyl-beta-D-glucopyranoside) hydrolytic activity was found in culture filtrate from Penicillium multicolor IAM7153, and the enzyme was isolated. The enzyme was purified as a beta-primeverosidase-like enzyme by precipitation with ammonium sulfate followed by successive chromatographies on Phenyl Sepharose, Mono Q, and beta-galactosylamidine affinity columns. The molecular mass was estimated to be 50 kDa by SDS-PAGE and gel filtration. The purified enzyme was highly specific toward the substrate p-nitrophenyl beta-primeveroside, which was cleaved in an endo-manner into primeverose and p-nitrophenol, but a series of beta-primeveroside as aroma precursors were hydrolyzed only slightly as substrates for the enzyme. In analyses of its hydrolytic action and kinetics, the enzyme showed narrow substrate specificity with respect to the aglycon and glycon moieties of the diglycoside. We conclude that the present enzyme is a kind of beta-diglycosidase rather than beta-primeverosidase.

Chromatography, Affinity↗
PubMedDate not supplied

Multicolor karyotype analyses of mouse embryonic stem cells.

The manipulation of embryonic stem (ES) cells to introduce directional genetic changes into the genome of mice has become an important tool in biomedical research. Monitoring of cell morphology before and after DNA manipulation and special culture conditions are a prerequisite to preserve the pluripotent properties of ES cells and thus their ability to generate chimera and effective germline transmission (GLT). It has been reported that prolonged cell culturing may affect the diploid chromosomal composition of cells and therefore the percentage of chimerism and GLT. Herein, we report multicolor-fluorescence in situ hybridization (M-FISH) analysis of four different ES cell lines/clones. Although the morphology of all four ES cell lines/clones appeared normal and all four expressed the early markers Oct-3/4 and Nanog, two cell lines presented consistent numerical and structural chromosome aberrations. We demonstrate that M-FISH is a sensitive and accurate method for a comprehensive karyotype analysis of ES cells and may minimize time, costs, and disappointments due to inadequate ES cell sources.

Animals↗
PubMed2003

bcl-2 expression by multicolor flow cytometric analysis assists in the diagnosis of follicular lymphoma in lymph node and bone marrow.

The expression of bcl-2, CD10, and CD20 was examined by multicolor flow cytometry in 78 samples including lymph node or other tissue biopsy specimens containing follicular lymphoma (FL; n = 17), reactive hyperplasia (RH; n = 28), or other malignant lymphomas (n = 20), as well as bone marrow aspirates (n = 13). The presence of CD10+ cells with high bcl-2 expression predicted the presence of FL rather than RH with a positive predictive value of 100% and negative predictive value of 96%. CD10+ cells with high bcl-2 expression also were found in a subset of diffuse large B-cell lymphomas and were otherwise rare in other types of malignant lymphoma. In contrast with immunohistochemical studies, a reduced but apparently measurable level of bcl-2 was present in benign follicular center cells. Hematogones showed lower bcl-2 levels than did FL cells in the bone marrow, and neutrophils were bcl-2-. Measurement of bcl-2 expression levels by multiparameter flow cytometry offers a rapid, quantitative assessment that may assist in the diagnosis of FL in lymph nodes or bone marrow, even when other CD10+ cells or admixed normal B cells are present.

Bone Marrow↗
PubMed2005

Prism-based excitation wavelength selection for multicolor fluorescence coincidence measurements.

We have designed and constructed a prism setup for multiple-color confocal fluorescence experiments. The prism setup permits easy selection of any color or any combination of colors from a multicolor light source, such as a mixed-gas argon-krypton-ion laser. The selected colors emerging from the prism setup are, by design, optimally overlapped in the focus of a high-numerical-aperture objective, such as that commonly used in single-molecule fluorescence experiments. The various excitation powers can be easily adjusted in this setup. We will exemplify the potential of this setup in single-molecule fluorescence emission spectroscopy.

Journal Article↗
PubMed2004

Multicolor lattice solitons.

We report on the existence of multicolor solitons supported by periodic lattices made from quadratic nonlinear media. Such lattice solitons bridge the gap between continuous solitons in uniform media and discrete solitons in strongly localized systems and exhibit a wealth of new features. We discovered that, in contrast to uniform media, multipeaked lattice solitons are stable. Thus they open new opportunities for all-optical switching based on soliton packets.

Journal Article↗
PubMed2004

Packing, unpacking, and steering of multicolor solitons in optical lattices.

We discuss potential applications of multicolor solitons supported by periodic lattices imprinted in quadratic nonlinear media. Such lattice solitons can be packed together with appropriate relative phases to form stable soliton trains that can be treated as bit sequences. We describe controllable splitting of the trains into their soliton constituents and the angle- and power-controlled steering and trapping of solitons moving across the lattice into the desired guiding channel.

Journal Article↗
PubMed2005

Analysis and synthesis of multicolored objects in a single image.

We present a method with which to recover the intrinsic shading and reflectance characteristics of multicolored three-dimensional objects in a single image, with which realistic new scenes can be synthesized. A color watershed algorithm, which is based on a regularized dichromatic fitting error, is proposed for robust image segmentation. For shading recovery in small regions, a weighted interpolation is employed, whereas in large regions the reflectance and shading are calculated based on the assumption of gradual shape variation. It is demonstrated that the proposed method is promising and can be applied in image simulation.

Journal Article↗
PubMed2006

Generation of multicolor vector Kerr solitons by cross-phase modulation, four-wave mixing, and stimulated Raman scattering.

We numerically and experimentally show the existence of multicolor vector spatial solitons in a Kerr planar waveguide through the combined effects of cross-phase modulation, four-wave mixing, and stimulated Raman scattering. Mutual spatial guiding of the Raman-Stokes, anti-Stokes, and pump waves is achieved in the high-conversion regime mainly by cross-phase modulation and phase-matched four-wave mixing induced by a power imbalance between Stokes and anti-Stokes components, leading to the generation of a clear-cut sech-shape three-frequency spatial soliton.

Journal Article↗
PubMed2005

Multicolor spectral karyotyping in multiple myeloma.

Multiple myeloma is a plasma cell disorder characterized at the cytogenetic level by aneuploid karyotypes with numerous complex structural aberrations. Unfortunately, conventional chromosome-banding techniques are unable to resolve many of these aberrations. Multicolor Spectral Karyotyping (SKY) is a molecular cytogenetic technique that utilizes chromosome-painting probes for characterizing complex structural chromosome aberrations. This innovative technology allows the simultaneous display of each of the 24 different chromosomes in a different color. Chromosomal sites of rearrangement or translocation between different chromosomes are visualized by a change in the color at the point of the aberration. The SKY technique makes possible the identification of chromosomal bands of unknown origin, including trans locations, insertions, complex rearrangements, and marker chromosomes, which, in many cases, go unresolved by traditional banding methods.

Bone Marrow Cells↗
PubMed2002

Purification and characterization of a laccase from the white-rot fungus Trametes multicolor.

The wood-degrading fungus Trametes multicolor secretes several laccase isoforms when grown on a simple medium containing copper in the millimolar range for stimulating laccase synthesis. The main isoenzyme laccase II was purified to apparent homogeneity from the culture supernatant by using anion-exchange chromatography and gel filtration. Laccase II is a monomeric glycoprotein with a molecular mass of 63 kDa as determined by sodium dodecylsulfate polyacrylamide gel electrophoresis, contains 18% glycosylation, and has a pI of 3.0. It oxidizes a variety of phenolic substrates as well as ferrocyanide and iodide. The pH optimum depends on the substrate employed and shows a bell-shaped pH activity profile with an optimum of 4.0 to 5.0 for the phenolic substrates, while the nonphenolic substrates ferrocyanide and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonate) show a monotonic pH profile with a rate decreasing with increasing pH.

Basidiomycota↗
PubMed2005

Toxicity of commonly used insecticides in sweet corn and soybean to multicolored Asian lady beetle (Coleoptera: Coccinellidae).

Use of insecticides with low toxicity to natural enemies is an important component of conservation biological control. In this study, we evaluated the toxicity of insecticides used in sweet corn, Zea mays L., and soybean, Glycine max (L.) Merr., to the multicolored Asian lady beetle, Harmonia axyridis (Pallas), under laboratory and field conditions. Field experiments conducted in sweet corn in 2003 and 2004 and in soybean in 2003, showed that H. axyridis was the most abundant predator. In sweet corn, densities of H. axyridis larvae in plots treated with spinosad or indoxacarb were generally higher than in plots treated with chlorpyrifos, carbaryl, bifenthrin, and A-cyhalothrin. In soybean, densities of H. axyridis larvae in plots treated with chlorpyrifos were higher than in plots treated with lambda-cyhalothrin. Laboratory experiments were conducted to evaluate the acute toxicity of insecticides to eggs, first and third instars, pupae, and adults. Spinosad, followed by indoxacarb, were the least toxic insecticides for all life stages of H. axyridis. Conventional insecticides showed high toxicity to H. axyridis when applied at field rates under laboratory conditions. Overall, first instars were most susceptible to the insecticides tested, followed by third instars and adults, eggs, and pupae. Our results suggest that spinosad, and to a lesser extent indoxacarb, offer reduced toxicity to H. axyridis and would be beneficial for conservation biological control in agricultural systems where H. axyridis is abundant.

Animals↗
PubMed2006

Application of multicolor banding for identification of complex chromosome 18 rearrangements.

Multicolor chromosome banding (mBAND) is a recently developed technique that allows the delineation of chromosomal regions with a resolution of a few megabase pairs. The resolution of mBAND is slightly below that of conventional chromosome banding; however, the color bands have a great value in identifying chromosomal abnormalities, particularly complex chromosome rearrangements, and intrachromosome exchanges (ie, inversions, deletions, duplications, and insertions). These abnormalities cannot be defined easily by conventional cytogenetic analysis or chromosome paint. In this report, we present the application of the mBAND analysis for identification of complex intrachromosome rearrangements of chromosome 18 in a child with dysmorphic features.

Chromosome Aberrations↗
PubMed1999

Role of multicolor fluorescence in situ hybridization (FISH) in simultaneous detection of probe sets for chromosome 18, X and Y in uncultured amniotic fluid cells.

Major aneuploidies diagnosed prenatally involve the autosomes 13, 18, and 21, and sex chromosomes. Fluorescence in situ hybridization (FISH) allows rapid analysis of chromosome copy number in interphase cells. The purpose of this study was to evaluate the role of multicolor fluorescence in situ hybridization in simultaneous detection of probe sets for chromosome 18, X, and Y in uncultured amniotic fluid cells as a safer alternative method for aneuploidy detection prenatally. Fifty amniotic fluid samples were analyzed by FISH and standard cytogenetics. Mean time to obtain results was three days for fluorescence in situ hybridization and 20 days for karyotype. Fluorescence in situ hybridization was informative in 43 samples (86%), and within this group, two aneuploidies were correctly identified. This evaluation demonstrates that FISH with X, Y, and 18 alpha satellite DNA probes could accurately and rapidly detect aneuploidies involving these chromosomes and could be used in any prenatal clinical laboratory.

Amniocentesis↗
PubMed2000

Multicolor spectral karyotype analysis of a transplantable human ileal carcinoid.

In this report we present the results of a combined cytogenetic and multicolor spectral karyotype (SKY) analysis of a transplantable human ileal carcinoid (GOT1). By using SKY it was possible to identify the origin and organization of all clonal marker chromosomes and to identify cryptic translocations not detectable by conventional chromosome banding. The stemline karyotype of low passage GOT1 cells was interpreted as 43,XX, der(1)del(1)(?), inv(2)(p25q13), del(3)(p21), del(5)(q13q31), del(6)(q13), -9, -13, -15, del(16) (q22). Analysis of the GOT1 cells after about 2.5 years of propagation in nude mice allowed us to follow the in vivo progression of this tumor. Relatively few additional rearrangements had occurred during this period, indicating that the GOT1 cells are genetically stable. Most of the abnormalities detected result in loss of whole or parts of chromosomes, suggesting that loss of multiple chromosomal regions, presumably containing tumor suppressor genes, might be important genetic events in ileal carcinoids.

Animals↗