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At least 469 records · Page 26Linked to original sources
PubMed1999

Evaluation of micronuclei and chromosomal breakage in the 1cen-q12 region by the butadiene metabolites epoxybutene and diepoxybutane in cultured human lymphocytes.

1,3-Butadiene is a widely used industrial chemical and common environmental pollutant that has been associated with increased risks of leukemias and lymphomas. Butadiene and its metabolites, 1, 2-epoxybutene (EB) and diepoxybutane (DEB), have been shown to be genotoxic in a wide variety of test organisms. The objective of this research was to evaluate techniques for the rapid detection of chromosomal alterations occurring in humans exposed to butadiene. We have used a multicolored fluorescence in situ hybridization (FISH) method and the CREST-modified micronucleus assay to detect chromosomal breakage induced by EB (10-300 microM) and DEB (0.5-10 microM) in cultured human lymphocytes. A significant dose-related increase in the formation of micronuclei was seen in lymphocytes treated with DEB at concentrations as low as 2.5 microM, but not with EB over the dose range tested. Over 80% of the micronuclei induced by DEB were CREST-negative, indicating their origin from chromosomal breakage. Multicolor FISH using two adjacent chromosome-specific probes showed a significant increase in chromosomal breakage in the 1cen-q12 region induced by DEB at concentrations as low as 2.5 microM, but not by EB. Since DEB is likely to be one of the metabolites contributing to the genotoxic effects of butadiene, the sensitivity of the tandem FISH approach to detect breakage induced by diepoxybutane indicates that this technique may be useful for monitoring chromosomal alterations in butadiene-exposed workers.

Butadienes↗
PubMed1988

Nipple discharge: surgical significance.

Nipple discharge is an important clinical entity ranking second only to a lump as the most common complaint among 7,588 patients having breast surgery (560/7,588, or 7.4%). In the office and clinic it is even more common, since many patients can be treated medically and do not require an operation. To be significant, a discharge should be true, spontaneous, persistent, and nonlactational. Nipple discharge can be milky, multicolored and sticky, purulent, clear (watery), yellow (serous), pink (serosanguineous), or bloody (sanguineous). Watery, serous, serosanguineous, and sanguineous discharges are surgically significant; while they are most often caused by intraductal papillomas or fibrocystic disease, they can be due to cancer or a precancerous mastopathy. Among 503 patients operated on for one of these types of discharge, 67 (13.3%) had cancer, and 36 (7.2%) had a precancerous mastopathy. Among the 67 patients with cancer, eight (11.9%) had no palpable mass, 11 (16.4%) had negative cytologic findings, and seven (10.4%) had a negative mammogram. The incidence of associated cancers increases when the discharge is, in order of increasing frequency, serous, serosanguineous, sanguineous, or watery, when it is accompanied by a lump, when it is unilateral and from a single duct, when there are positive cytologic or mammographic findings, and when the patient is more than 50 years of age. Milky discharge caused by galactorrhea is treated medically except when caused by a pituitary adenoma. Multicolored sticky discharge due to duct ectasia is also treated medically except in advanced cases. Purulent discharge caused by an abscess requires drainage and a biopsy of the abscess wall. Except in women less than 30 years of age of in those anxious to have children, we advise a complete central duct excision for patients with surgically significant types of discharge. If done carefully, this procedure can yield good cosmetic results.

Adult↗
PubMed2004

Dynamic accommodation responses to stationary colored targets.

PURPOSE: When the targets or the background in a display are different colors, longitudinal chromatic aberration ensures that there is no single correct accommodative response. The purpose of the present study was to determine whether the response becomes more variable when viewing certain multicolor displays. METHODS: Accommodative responses of five young participants were measured with a dynamic infrared optometer while they viewed steady targets at a nominal stimulus level of 3 D. Target-on-background color combinations were black on white, black on blue, black on red, blue on red, red on blue, dark blue on red, and dark red on blue. RESULTS: When compared with the standard black-on-white target, responses to targets with reduced spectral bandwidth were not significantly more variable. In most participants, responses to near-isoluminant targets (e.g., red on blue and blue on red) were not more variable than to the standard target. However, calculated confidence intervals cannot rule out moderate to large changes in variability near isoluminance. Responses to these multicolor targets tended to favor the blue focus. CONCLUSIONS: In most individuals, viewing multichromatic targets does not increase significantly the variation in accommodative response as compared with broadband black-and-white targets.

Accommodation, Ocular↗
PubMed2005

Separating reflection components of textured surfaces using a single image.

In inhomogeneous objects, highlights are linear combinations of diffuse and specular reflection components. A number of methods have been proposed to separate or decompose these two components. To our knowledge, all methods that use a single input image require explicit color segmentation to deal with multicolored surfaces. Unfortunately, for complex textured images, current color segmentation algorithms are still problematic to segment correctly. Consequently, a method without explicit color segmentation becomes indispensable and this paper presents such a method. The method is based solely on colors, particularly chromaticity, without requiring any geometrical information. One of the basic ideas is to iteratively compare the intensity logarithmic differentiation of an input image and its specular-free image. A specular-free image is an image that has exactly the same geometrical profile as the diffuse component of the input image and that can be generated by shifting each pixel's intensity and maximum chromaticity nonlinearly. Unlike existing methods using a single image, all processes in the proposed method are done locally, involving a maximum of only two neighboring pixels. This local operation is useful for handling textured objects with complex multicolored scenes. Evaluations by comparison with the results of polarizing filters demonstrate the effectiveness of the proposed method.

Algorithms↗
PubMed2002

Memory-based detection of task-irrelevant visual changes.

Colored grating patterns were presented to 8 participants in a passive oddball condition (standard, 87.5% and deviant, 12.5%, differing in their color). In the corresponding multicolor condition, grating patterns of eight different colors were presented, their probabilities set equal both to each other and to that of the deviant in the oddball task. Compared with the ERP response elicited by the standard stimulus, the deviant response was negatively displaced over posterior areas, the difference wave peaking at 136 ms. A similar negative wave was obtained when the ERP response to the deviant was compared with the ERP elicited by the same stimulus in the multicolor condition. This result rules out stimulus- (color-) specific refractoriness as a major factor in the generation of the deviance-related posterior negativity. The observed posterior negativity can therefore be regarded as a visual analog of the mismatch negativity (vMMN).

Adolescent↗
PubMed2006

Rapid flow cytometry method for quantitation of LFA-1-adhesive T cells.

Adhesion molecules are important for leukocyte endothelial attachment and migration to sites of inflammation. The LFA-1 (CD11a and CD18) integrin molecule is constitutively expressed on the T-cell surface. Following T-cell activation, a rapid conformational change of LFA-1 to an "adhesive" state occurs, allowing LFA-1 binding to intracellular cell adhesion molecule type 1 (ICAM-1)-expressing targets, such as antigen-presenting cells. For this study, a rapid flow cytometry method for the quantitation of LFA-1-adhesive T cells following activation was developed. Purified ICAM-1 was bound to 4.5-microm-diameter beads. Following peripheral blood mononuclear cell activation culture (phorbol myristate acetate and ionomycin), the cells were incubated with the ICAM-1 beads, which allowed attachment to occur. The T cell-bead complexes were then resolved from unbound T cells by flow cytometry. Multicolor analysis allowed a complete phenotypic analysis of the adhesive T-cell subsets. Experimental controls indicated that the T cell-bead attachment was LFA-1 and ICAM-1 specific. Very little binding between unactivated T cells and ICAM beads or between activated T cells and plain beads was observed. The kinetics of the response was extremely rapid, with nearly maximal numbers of adhesive T cells observed following 5 min of activation. Scanning electron microscopy analysis was used to characterize legitimate bead-cell binding. By using multicolor cytometry, the responding adhesive T-cell population was usually identified as a distinct subset of T cells with the following phenotype: CD3+ CD4+ or CD8+ CD19- CD16- CD45RO+ CD62L+ CD27+ CD57-. A rapid and simple method for the scoring of LFA-1-adhesive T cells was developed and may have significant utility for immune function studies.

Cell Adhesion↗
PubMed2005

Use of a novel panel of nine short tandem repeats for exact and high-resolution fingerprinting of Aspergillus fumigatus isolates.

Here we describe a new panel of short tandem repeats (STRs) for a novel exact typing assay that can be used to discriminate between Aspergillus fumigatus isolates. A total of nine STR markers were selected from available genomic A. fumigatus sequences and were divided into three multicolor multiplex PCRs. Each multiplex reaction amplified three di-, tri-, or tetranucleotide repeats, respectively. All nine STR markers were used to analyze 100 presumably unrelated A. fumigatus isolates. For each marker, between 11 and 37 alleles were found in this population. One isolate proved to be a mixture of at least two different isolates. With the remaining 99 isolates, 96 different fingerprinting profiles were obtained. The Simpson's diversity index for the individual markers ranged from 0.77 to 0.97. The diversity index for the multiplex combination of di-, tri-, and tetranucleotide repeats ranged from 0.9784 to 0.9968. The combination of all nine markers yielded a Simpson's diversity index of 0.9994, indicative of the high discriminatory power of these new loci. In theory, this panel of markers is able to discriminate between no less than 27 x 10(9) different genotypes. The multicolor multiplex approach allows large numbers of markers to be tested in a short period of time. The exact nature of the assay combines high reproducibility with the easy exchange of results and makes it a very suitable tool for large-scale epidemiological studies.

Aspergillus fumigatus↗
PubMed2005

Quantitative 3D fluorescence technique for the analysis of en face preparations of arterial walls using quantum dot nanocrystals and two-photon excitation laser scanning microscopy.

Traditional imaging with one-photon confocal microscopy and organic fluorophores poses several challenges for the visualization of vascular tissue, including autofluorescence, fluorophore crosstalk, and photobleaching. We studied human coronary arteries (HCAs) and mouse aortas with a modified immunohistochemical (IHC) "en face" method using quantum dot (Qdot) bioconjugates and two-photon excitation laser scanning microscopy (TPELSM). We demonstrated the feasibility of multilabeling intimal structures by exciting multicolored Qdots with only one laser wavelength (750 nm). Detailed cell structures, such as the granular appearance of von Willebrand factor (VWF) and the subcellular distribution of endothelial nitric oxide synthase, were visualized using green dots (525 nm), even when the emission maximum of these Qdots overlapped that of tissue autofluorescence (510-520 nm). In addition, sensitive fluorescence quantification of vascular cell adhesion molecule 1 expression at areas of varying hemodynamics (intercostal branches vs. nonbranching areas) was performed in normal C57Bl/6 mice. Finally, we took advantage of the photostability of Qdots and the inherent three-dimensional (3D) resolution of TPELSM to obtain large z-stack series without photobleaching. This innovative en face method allowed simple multicolor profiling, highly sensitive fluorescence quantitation, and 3D visualization of the vascular endothelium with excellent spatial resolution. This is a promising technique to define the spatial and temporal interactions of endothelial inflammatory markers and quantify the effects of different interventions on the endothelium.

Animals↗
PubMedDate not supplied

First case of trisomy 13 plus mosaic trisomy 1q.

OBJECTIVES: In a case with severe sonographic abnormalities followed by missed abortion in week 14 + 5 days, cytogenetic analysis was performed on placental tissue. A mosaic karyotype 47,XY,+13,add(1)(q44)[3]/47,XY, +13[9]/46,XY[36] was detected. The purpose of the present study was to characterize the additional material on chromosome 1q. METHODS: According to GTG banding, the additional material on chromosome 1 was most probably chromosome 1 material. Thus, multicolor banding analysis using a chromosome 1 specific probe set was done to precisely describe the rearranged chromosome 1. RESULTS: Molecular cytogenetic approaches revealed that the derivative chromosome 1 was der(1)(1p36.3-->1q44::1q12-->1q44). CONCLUSIONS: This is the first description of a case with a trisomy 13 plus a partial trisomy 1q presenting with clinical signs of both aberrations. Moreover, the multicolor banding technique is suited to resolve complex karyotypes in the prenatal diagnosis, i.e., chromosome preparations of fibroblasts.

Abortion, Missed↗
PubMed2001

Classifying by colors: FISH-based genome analysis.

In recent years a fascinating evolution of different multicolor fluorescence in situ hybridization (FISH) technologies could be witnessed. The various approaches to cohybridize multiple DNA probes in different colors opened new avenues for FISH-based automated karyotyping or the simultaneous analysis of multiple defined regions within the genome. These developments had a remarkable impact on microscopy design and the usage of highly sensitive area imagers. In addition, they led to the introduction of new fluorochromes with appropriate filter combinations, refinements of hybridization protocols, novel probe sets, and innovative software for automated chromosome analysis. This paper attempts to summarize the various multicolor approaches and discusses the application of the individual technologies.

Animals↗
PubMed2003

Seven-fluorochrome mouse M-FISH for high-resolution analysis of interchromosomal rearrangements.

The mouse has evolved to be the primary mammalian genetic model organism. Important applications include the modeling of human cancer and cloning experiments. In both settings, a detailed analysis of the mouse genome is essential. Multicolor karyotyping technologies have emerged to be invaluable tools for the identification of mouse chromosomes and for the deciphering of complex rearrangements. With the increasing use of these multicolor technologies resolution limits are critical. However, the traditionally used probe sets, which employ 5 different fluorochromes, have significant limitations. Here, we introduce an improved labeling strategy. Using 7 fluorochromes we increased the sensitivity for the detection of small interchromosomal rearrangements (700 kb or less) to virtually 100%. Our approach should be important to unravel small interchromosomal rearrangements in mouse models for DNA repair defects and chromosomal instability.

Animals↗
PubMed2006

Nuclear DNA content and nuclear and cell volume are positively correlated in angiosperms.

Volumes of flow sorted nuclei were analyzed from two highly endopolyploid (diploids with endopolyploid tissues) species (Arabidopsis thaliana and Barbarea stricta), from a less endopolyploid species (Allium cepa) and from two non-endopolyploid species (Chrysanthemum multicolor and Fritillaria uva-vulpis). Intraspecific as well as interspecific comparisons revealed a highly positive correlation (r > 0.99) between DNA content and nuclear volume. No significant differences between expected and measured nuclear volumes were noted indicating that chromatin packing is not increased with increasing DNA content in the tested plant species. In epidermis cells of A. thaliana, A. cepa and Ch. multicolor, a lower (r between 0.6 and 0.7) but significant positive correlation between nuclear volume and cell volume was found. This correlation is compatible with the hypothesis that endopolyploidization (EP = consecutive replication cycles not separated by nuclear and cell divisions) might speed up the growth of endopolyploid species and compensate for small genome size.

Arabidopsis↗
PubMed2006

Supernumerary marker chromosome 5 diagnosed by M-FISH in a child with congenital heart defect and unusual face.

We describe a female patient with a small supernumerary marker chromosome (sSMC) present in mosaic and characterized in detail by fluorescence in situ hybridization (FISH) using all 24 human whole chromosome painting probes, multicolor banding (MCB) and subcentromere specific multicolor FISH (subcenM-FISH). The sSMC was demonstrated to be derived from chromosome 5 and the karyotype of our patient was as follows: 47,XX,+mar.ish r(5)(::p13.2 approximately p13.3-->q11.2::) [60%]/46,XX [40%]. Partial trisomy for the proximal 5p and q chromosomal regions is a rare event. A critical region exists at 5p13 for the phenotype associated with duplication 5p. As far as we know, eight similar cases have been published up to now. We describe a new case which, to our knowledge, is the first characterized in such detail. The role of uniparental disomy (UPD) in cases of SMC is also discussed.

Abnormalities, Multiple↗
PubMed2006

De novostructural chromosomal imbalances: molecular cytogenetic characterization of partial trisomies.

De novo structural chromosomal imbalances represent a major challenge in modern cytogenetic diagnostics. Based solely on conventional cytogenetic techniques it may be impossible to identify the chromosomal origin of additional chromosomal material. In these cases molecular cytogenetic investigations including multicolor-FISH (M-FISH), spectral karyotyping (SKY), multicolor banding (MCB) and cenM-FISH combined with appropriate single-locus FISH probes are highly suitable for the determination of the chromosomal origin and fine characterization of derivative chromosomes. Here we report on four patients with de novo chromosomal imbalances and distinct chromosomal phenotypes, three of them harboring pure partial trisomies: a mildly affected boy with pure partial trisomy 10q22.2-->q22.3 approximately 23.1 due to an interstitial duplication, a girl with pure trisomy 12p11.21-->pter and atypically moderate phenotype as the consequence of an X;autosome translocation, and a girl with multiple congenital abnormalities and severe developmental delay and a 46,XX,15p+ karyotype hiding a trisomy 17pter-->17q11.1. The fourth patient is a girl with minor phenotypic features and mental retardation with an inverted duplication 18q10-->p11.31 combined with a terminal deletion of 18p32. The clinical pictures are compared with previously described patients with focus on long term outcome.

Chromosome Aberrations↗
PubMed2000

Seven-color fluorescence imaging of tissue samples based on Fourier spectroscopy and singular value decomposition.

Seven-color analyses of immunofluorescence-stained tissue samples were accomplished using Fourier spectroscopy-based hyperspectral imaging and singular value decomposition. This system consists of a combination of seven fluorescent dyes, three filtersets, an epifluorescence microscope, a spectral imaging system, a computer for data acquisition, and data analysis software. The spectra of all pixels in a multicolor image were taken simultaneously using a Sagnac type interferometer. The spectra were deconvolved to estimate the contribution of each component dye, and individual dye images were constructed based on the intensities of assigned signals. To obtain mixed spectra, three filter sets, i.e., Bl, Gr, and Rd for Alexa488 and Alexa532, for Alexa546, Alexa568, and Alexa594, and for Cy5 and Cy5.5, respectively, were used for simultaneous excitation of two or three dyes. These fluorophores have considerable spectral overlap which precludes their separation by conventional analysis. We resolved their relative contributions to the fluorescent signal by a method involving linear unmixing based on singular value decomposition of the matrices consisting of dye spectra. Analyses of mouse thymic tissues stained with seven different fluorescent dyes provided clear independent images, and any combination of two or three individual dye images could be used for constructing multicolor images.

Animals↗
PubMed1995

Fluorescence overlay antigen mapping of the epidermal basement membrane zone: II. Color fidelity.

In this second report on the fluorescence overlay antigen mapping (FOAM) technique, we highlight some of the errors that may influence faithful color rendition of slide preparations using triple antigen immunofluorescence staining. Reliable interpretation of multicolor fluorescence images requires that the observer can unambiguously assign each color in these images to the presence of a specific combination of the labeled antigens. This is possible only when the image fidelity meets certain standards. The present study concentrates on color fidelity which is easily undermined by spectral matching errors, image contrast errors, and exposure time errors. Evaluation of these errors, using the photomicrographic overlay variant of FOAM, showed the potential unreliability of the simultaneous use of multiple fluorophores for immunofluorescence microscopy. The procedures described here may serve as a solid starting point in formulating technical conditions that allow reliable color rendition in multicolor immunofluorescence microscopy. Furthermore, these procedures can be adapted to studies other than the analysis of basement membrane zone antigens, to which they have been first applied.

Basement Membrane↗
PubMed2006

Hidden chromosomal abnormalities in pleuropulmonary blastomas identified by multiplex FISH.

BACKGROUND: Pleuropulmonary blastoma (PPB) is a rare childhood dysontogenetic intrathoracic neoplasm associated with an unfavourable clinical behaviour. CASES PRESENTATION: We report pathological and cytogenetic findings in two cases of PPB at initial diagnosis and recurrence. Both tumors were classified as type III pneumoblastoma and histological findings were similar at diagnosis and relapse. In both cases, conventional cytogenetic techniques revealed complex numerical and structural chromosomal abnormalities. Molecular cytogenetic analysis (interphase/metaphase FISH and multicolor FISH) identified accurately chromosomal aberrations. In one case, TP53 gene deletion was detected on metaphase FISH. To date, only few cytogenetic data have been published about PPB. CONCLUSION: The PPB genetic profile remains to be established and compared to others embryonal neoplasia. Our cytogenetic data are discussed reviewing cytogenetics PPBs published cases, illustrating the contribution of multicolor FISH in order to identify pathogenetically important recurrent aberrations in PPB.

Child, Preschool↗
PubMed2002

Compact red-green-blue beam illuminator and expander.

A compact multicolor beam illuminator and a compact multicolor beam expander are presented. The illuminator performs the dual task of demultiplexing a narrow red-green-blue (RGB) input beam into three separate beams, each of a different color, and expanding them to yield three separate magnified plane waves. The beam expander expands a narrow RGB input beam into a single magnified RGB plane wave. The design and recording procedures, along with experimental results for a beam illuminator and a beam expander with a magnification of approximately 3, are presented.

Journal Article↗