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At least 1,063 records · Page 59Linked to original sources
PubMed2000

Galactosylation of thiol group by beta-galactosidase.

beta-Galactosidase catalyzed beta-galactosylation not only of a hydroxyl group but also of a thiol group in the condensation reaction of D-galactose and 2-mercaptoethanol. The thio-galactosylation product was confirmed as 2-hydroxyethyl S-beta-D-galactoside on the bases of fast atom bombardment mass spectrometry, infrared spectroscopy, and nuclear magnetic resonance spectorometry. Aspergillus oryzae beta-galactosidase hydrolyzed p-nitrophenyl S-beta-D-galactoside most rapidly among several beta-galactosidases and produced the thio-galactosylation product most efficiently. The Penicillim multicolor enzyme was as effective as the A. oryzae enzyme. However the enzymes from Escherichia coli, Saccharomyces fragilis, Kluyveromyces lactis, and Bacillus circulans galactosylated hydroxyl groups predominantly to produce O-galactoside. The thio-galactoside was synthesized most effectively at a 2-mercaptoethanol concentration of about 1.25 M. Galactose concentration at 0.8-2.8 M did not affect the synthetic yield of the thiogalactoside so greatly.

Aspergillus oryzae↗
PubMed2001

Organophosphorous pesticide exposure increases the frequency of sperm sex null aneuploidy.

It has been estimated that 4 of 1,000 live births and 35% of spontaneous abortions are aneuploid and that an important proportion of embryo and newborn aneuploidy is of paternal origin. Exposure to organophosphorous pesticides (OP) has been associated with sperm hyperploidy/polyploidy. Therefore, we aimed to assess the frequency of sperm aneuploidy (X, Y, and 18) and its relationship with urinary OP metabolites in agricultural workers. We performed multicolor fluorescence in situ hybridization on samples from nine men obtained before and during the pesticide spraying season to assess sperm aneuploidy. We measured urinary OP metabolite levels by gas-liquid chromatography. Aneuploidies were found in 0.67% of total sperm nuclei. The most frequent aneuploidy was the lack of a sexual chromosome or sex null (0.19%), followed by XY18 (0.15%) and XY18-18 (0.06%). OP metabolites detected at higher concentrations were dimethylthiophosphate, dimethyldithiophosphate, and diethylphosphate (DEP). There were no differences in average aneuploidy frequency or urinary metabolite levels between samples collected before and after exposure. However, Poisson regression analysis adjusted for age, alcohol intake, and sperm concentration showed significant associations between OP metabolite concentrations and increased frequency of sperm aneuploidies. The association was more evident between DEP and sex null, and the risk increased further during the spraying season. Thus, OP exposure could interfere with sperm chromosome segregation and increase the risk for genetic syndromes, such as Turner's. Further studies are required to assess the prevalence of spontaneous abortions, birth defects, and genetic syndromes in agricultural communities.

Abortion, Spontaneous↗
PubMed1996

Detection of aneuploidy in human spermatozoa of normal semen donors by fluorescence in situ hybridization.

We have studied human spermatozoa from 24 normal, healthy unexposed men, 18 of whom were semen donors at the Sperm Bank in Turku, using multicolor fluorescence in situ hybridization with two chromosome-specific probes. The possible age-related increase in aneuploidy frequencies was assessed. Ten thousand spermatozoa were scored per individual for the presence of hyperploid, i.e., disomic and diploid, cells. The overall hybridization efficiency was 98.8%. The frequency of spermatozoa with two chromosome 1 signals was 11.5 +/- 5.2/10,000. The frequency of spermatozoa with two chromosome 7 signals was 6.4 +/- 3.9/10,000. Diploidy was present in 15.0 +/- 8.9/10,000 spermatozoa. Interindividual variation was quite large. No statistically significant correlation between age of the donors (range = 20-46 years) and the frequency of hyperploid spermatozoa was observed. The results give background information on the incidence of hyperploid spermatozoa in unexposed men and encourage the use of this novel technique of future studies on genetic effects in men exposed to potentially aneuploidogenic agents.

Adult↗
PubMed2001

Population study and validation of paternity testing for Thoroughbred horses by 15 microsatellite loci.

Microsatellite 15 TKY System was characterized for parentage verification of horse registry. The Microsatellite 15 TKY System was constructed by using 15 microsatellites, TKY279, TKY287, TKY294, TKY297, TKY301, TKY312, TKY321, TKY325, TKY333, TKY337, TKY341, TKY343, TKY344, TKY374, and TKY394, to provide stringent PCR-based microsatellite typing specifically optimized for multicolor fluorescence detection. The Microsatellite 15 TKY System showed good resolutions for 250 unrelated Thoroughbred horses, and the probability of exclusion (PE) at each microsatellite ranged from 0.437 to 0.621, resulting in a total PE value of 99.998% for Thoroughbred horses. These results indicated that the Microsatellite 15 TKY System is useful for paternity testing of Thoroughbred horses. A paternity testing case for a Thoroughbred horse family, in which candidate sires had close relations, was analyzed using the Microsatellite 15 TKY System. In this case, the Microsatellite 15 TKY System excluded paternity of a false sire. We concluded that the Microsatellite 15 TKY System can give sufficient and reliable information for paternity testing.

Animals↗
PubMed2003

Detection of mature T-cell leukemias by flow cytometry using anti-T-cell receptor V beta antibodies.

A broad array of antibodies directed against the variable (V) region of the T-cell receptor (TCR) beta (V beta) chain has become available in a directly conjugated multicolor format that permits assessment of 19 of 25 V beta families, covering 70% of the normal circulating T-cell repertoire. These antibodies were used to detect expanded T-cell populations in 43 peripheral blood samples submitted for suspected T-cell malignancy. Of 43 samples, 27 were diagnosed as follows: T-cell large granular lymphocyte leukemia, 14 samples; Sézary syndrome, 4 samples; T-cell prolymphocytic leukemia, 5 samples; or T-cell non-Hodgkin lymphoma or T-cell lymphoproliferative disorder not otherwise specified, 4 samples. The remaining 16 samples were determined to be nonneoplastic. All samples were diagnosed before assessment with anti-V beta flow cytometry. By using a cutoff of 1.6 times the upper limit of normal range (ULN) to define malignant restriction of V beta use, pathologic restriction of V beta use was found directly or indirectly in all 27 samples carrying a diagnosis of malignancy and directly in 2 of 16 samples without a diagnosis of malignancy. TCR gene rearrangement studies were used to confirm V beta flow cytometry results. By using a cutoff of 1.6 times the ULN for the detection of malignancy, the antibody panel had a diagnostic sensitivity of 89% for direct detection of pathologic V beta restriction and a specificity of 88%, making it useful for rapid diagnosis of T-cell leukemia.

Antibodies↗
PubMed2004

Post-PCR multiplex fluorescent ligation detection assay and flow cytometry for rapid detection of gene-specific translocations in leukemia.

We describe a novel method to detect specific polymerase chain reaction (PCR) target amplicons, involving thermostable ligation of fluorescent and biotinylated oligonucleotides, microparticle bead capture of the ligated products, and flow cytometric analysis. This approach, termed fluorescent ligation detection reaction (f-LDR) is more rapid and cost-effective than oligoprobe Southern blot hybridization (SBH). A standard f-LDR protocol was developed to detect the leukemia-associated chimeric transcripts bcr-abl and promyelocytic leukemia-retinoic acid receptor a (PML-RARalpha) in 2 multiplex and multicolor assays. The f-LDR platform was 100% specific and demonstrated comparable or better sensitivity than standard oligoprobe SBH. The usefulness of f-LDR was evaluated in 94 posttherapy samples from 13 patients with acute promyelocytic leukemia with the PML-RARalpha gene fusion. The f-LDR method was highly concordant (93%) with oligoprobe SBH; essentially all discrepancies were noted to be due to the enhanced sensitivity of f-LDR. We conclude that f-LDR is a highly specific and sensitive post-PCR method with wide potential application.

Cell Line, Tumor↗
PubMed2002

Full-color imaging through a turbid medium by use of photorefractive coherence gating and a technique to separate the recording spaces.

We demonstrate full-color imaging through a turbid medium by use of photorefractive coherence gating and a technique to separate the recording space of each color from those of the other colors. We found that the recording spaces must be separate when a multicolor image is recorded in a photorefractive crystal to prevent the interference of the holograms with one another. For full-color imaging we used a He-Cd white-light laser, which is compact and useful for full-color holography. Full-color-image retrieval is demonstrated through five mean free paths of a turbid medium.

Journal Article↗
PubMed2002

Technique for phase measurement and surface reconstruction by use of colored structured light.

We present a new method for improving the measurement of three-dimensional (3-D) shapes by using color information of the measured scene as an additional parameter. The widest used algorithms for 3-D surface measurement by use of structured fringe patterns are phase stepping and Fourier fringe analysis. There are a number of problems and limitations inherent in these algorithms that include: that the phase maps produced are wrapped modulo 2pi, that in some cases the acquired fringe pattern does not fill the field of view, that there may be spatially isolated areas, and that there is often invalid and/or noisy data. The new method presented to our knowledge for the first time here uses multiple colored fringe patterns, which are projected at different angles onto the measured scene. These patterns are analyzed with a specially adapted multicolor version of the standard Fourier fringe analysis method. In this way a number of the standard difficulties outlined above are addressed.

Journal Article↗
PubMed2005

Transferring color between three-dimensional objects.

A framework for transferring image-based color between three-dimensional objects by the use of a dichromatic reflection model is proposed. The framework addresses the following issues: (1) accurate recovery of an implicit geometric coefficient, (2) calculation of body color, (3) color transfer between different illuminants, and (4) segmentation of multicolored regions. The experimental results show that high color accuracy and photorealistic effects of the synthesized images can be achieved. The proposed technique has wide applications in image-based design and visualization of three-dimensional objects.

Algorithms↗
PubMed2005

Single-cavity, multiline laser system with large coherence length.

We present a new approach of generating the several wavelengths required for color holography with coherence lengths in the range of several meters. Our proposed laser system consists of an argon-ion laser, which is equipped with broadband optics. Its main lasing lines include 457, 488, and 514 nm. Sufficient coherence length is achieved by means of an intracavity etalon. We report single-frequency operation at several competing wavelengths and the successful recording of multicolor holograms with the described laser system.

Journal Article↗
PubMed2000

Color from shape from color: a simple formalism with known light sources

Photometric stereo is a well-known technique for recovering surface normals of a surface but requires three or more images of a surface taken under illumination from different directions. At best, one may dispense with the need for multiple images by using colored lights tuned to camera filters. But a less restrictive paradigm is available that uses the orientation-from-color approach, wherein multiple broadband illuminants impinge on a surface simultaneously. In that method, colors for a Lambertian surface lie on an ellipsoid in color space. The method has been applied mainly to single-color objects, with ellipsoid quadratic-form parameters determined from a large number of pixels. However, recently Petrov and Antonova [Color Res. Appl. 21, 97 (1996)] developed an entirely local approach, useful also for multicolored objects with color uniform in each patch. We investigate to what extent a method such as that of Petrov and Antonova can be applied in the ostensibly simpler situation in which the complex lighting environment is known, i.e., a color photometric stereo situation, with all lights in play at once with only a single image to analyze. We find that, assuming a simple model of color formation, we are able to recover the object colors along with surface normals, using only a single image. Because we immerse the object in a known lighting environment, we show that only half of the equations utilized by Petrov and Antonova are actually needed, making the method more stable. Nevertheless, solutions do not exist at every pixel; instead we may determine a best estimate of patch color, using a robust estimator, and then apply that estimate throughout a patch. Results are shown to be quite good compared with ground truth. The simple color model can often be made to hold more exactly by transforming the color space into one corresponding to spectrally sharpened sensors, which are a matrix transform away from the actual camera sensors. In our study the reliability and accuracy of the normal vector and of the surface color recovery algorithm are improved by this straightforward transformation.

Journal Article↗
PubMed2004

Color constancy through inverse-intensity chromaticity space.

Existing color constancy methods cannot handle both uniformly colored surfaces and highly textured surfaces in a single integrated framework. Statistics-based methods require many surface colors and become error prone when there are only a few surface colors. In contrast, dichromatic-based methods can successfully handle uniformly colored surfaces but cannot be applied to highly textured surfaces, since they require precise color segmentation. We present a single integrated method to estimate illumination chromaticity from single-colored and multicolored surfaces. Unlike existing dichromatic-based methods, the proposed method requires only rough highlight regions without segmenting the colors inside them. We show that, by analyzing highlights, a direct correlation between illumination chromaticity and image chromaticity can be obtained. This correlation is clearly described in "inverse-intensity chromaticity space," a novel two-dimensional space that we introduce. In addition, when Hough transform and histogram analysis is utilized in this space, illumination chromaticity can be estimated robustly, even for a highly textured surface.

Journal Article↗
PubMed2006

Pixel resolution control in numerical reconstruction of digital holography.

A new method for resolution control in numerical reconstruction of digital holography is proposed. The wave field on a tilted or vertical plane can be reconstructed without being subject to the minimum object-to-hologram distance requirement, and the pixel resolution can be easily controlled by adjusting the position of a transitional plane. The proposed method solves the problem of pixel resolution control for small object-to-hologram distances and is especially useful for multicolor, multiwavelength digital holography and metrological applications. Experimental results are presented to verify the idea.

Algorithms↗
PubMed2005

Evidence for high frequency of chromosomal mosaicism in spontaneous abortions revealed by interphase FISH analysis.

Numerical chromosomal imbalances are a common feature of spontaneous abortions. However, the incidence of mosaic forms of chromosomal abnormalities has not been evaluated. We have applied interphase multicolor fluorescence in situ hybridization using original DNA probes for chromosomes 1, 9, 13, 14, 15, 16, 18, 21, 22, X, and Y to study chromosomal abnormalities in 148 specimens of spontaneous abortions. We have detected chromosomal abnormalities in 89/148 (60.1%) of specimens. Among them, aneuploidy was detected in 74 samples (83.1%). In the remaining samples, polyploidy was detected. The mosaic forms of chromosome abnormality, including autosomal and sex chromosomal aneuploidies and polyploidy (31 and 12 cases, respectively), were observed in 43/89 (48.3%) of specimens. The most frequent mosaic form of aneuploidy was related to chromosome X (19 cases). The frequency of mosaic forms of chromosomal abnormalities in samples with male chromosomal complement was 50% (16/32 chromosomally abnormal), and in samples with female chromosomal complement, it was 47.4% (27/57 chromosomally abnormal). The present study demonstrates that the postzygotic or mitotic errors leading to chromosomal mosaicism in spontaneous abortions are more frequent than previously suspected. Chromosomal mosaicism may contribute significantly to both pregnancy complications and spontaneous fetal loss.

Abortion, Spontaneous↗
PubMed2005

The variation of aneuploidy frequency in the developing and adult human brain revealed by an interphase FISH study.

Despite the lack of direct cytogenetic studies, the neuronal cells of the normal human brain have been postulated to contain normal (diploid) chromosomal complement. Direct proof of a chromosomal mutation presence leading to large-scale genomic alterations in neuronal cells has been missing in the human brain. Large-scale genomic variations due to chromosomal complement instability in developing neuronal cells may lead to the variable level of chromosomal mosaicism probably having a substantial effect on brain development. The aim of the present study was the pilot assessment of chromosome complement variations in neuronal cells of developing and adult human brain tissues using interphase multicolor fluorescence in situ hybridization (mFISH). Chromosome-enumerating DNA probes from the original collection (chromosomes 1, 13 and 21, 18, X, and Y) were used for the present pilot FISH study. As a source of fetal brain tissue, the medulla oblongata was used. FISH studies were performed using uncultured fetal brain samples as well as organotypic cultures of medulla oblongata tissue. Cortex tissues of postmortem adult brain samples (Brodmann area 10) were also studied. In cultured in vitro embryonic neuronal brain cells, an increased level of aneuploidy was found (mean rate in the range of 1.3-7.0% per individual chromosome, in contrast to 0.6-3.0% and 0.1-0.8% in uncultured fetal and postmortem adult brain cells, respectively). The data obtained support the hypothesis regarding aneuploidy occurrence in normal developing and adult human brain.

Aneuploidy↗
PubMed2005

Three-dimensional imaging of the intracellular localization of growth hormone and prolactin and their mRNA using nanocrystal (Quantum dot) and confocal laser scanning microscopy techniques.

Semiconductor nanocrystals (Quantum dots, Qdots) have recently been used in biological research, because they do not fade on exposure to light, and they enable us to obtain multicolor imaging because of a narrow emission peak that can be excited via a single wavelength of light. There have been no reports of simultaneous localization of mRNA and protein using Qdots. We successfully applied these advantages of Qdot and confocal laser scanning microscopy (CLSM) to three-dimensional images of the intracellular localization of growth hormone and prolactin and to their mRNA. In situ hybridization and immunohistochemistry using Qdots combined with CLSM can optimally illustrate the relationship between protein and mRNA simultaneously in three dimensions. Such an approach enables us to visualize functional images of proteins in relation with mRNA synthesis and localization.

Animals↗
PubMed2005

Prenatal diagnosis and molecular cytogenetic characterization of an unusual complex structural rearrangement in a pregnancy following intracytoplasmic sperm injection (ICSI).

We report on a balanced complex chromosomal aberration detected in a fetus after amniocentesis. The pregnancy was achieved after intracytoplasmic sperm injection. GTG-banding revealed a complex structurally rearranged karyotype with a translocation between chromosomes 5 and 15 and an additional paracentric inversion in the der(15) between bands 5q11.2 and 5q15. Ag-NOR staining showed an interstitial active nuclear organizer region in the der(15). Molecular cytogenetic analyses using whole-chromosome-painting probes, comparative genomic hybridization, and multicolor banding did not point to further structural aberrations or imbalances. Therefore, a complex rearrangement with three breakpoints has occurred, and the karyotype can be described as 46,XX,der(5)t(5;15) (q11.2;p12),der(15)t(5;15)(q11.2;p12)inv(5)(q11.2q15).

Adult↗
PubMed2005

Another small supernumerary marker chromosome (sSMC) derived from chromosome 2: towards a genotype/phenotype correlation.

Here we report a prenatally detected small supernumerary marker chromosome (sSMC) derived from chromosome 2 as demonstrated by cenM-FISH (centromere-specific multicolor fluorescence in situ hybridization). By application of a recently described subcentromere-specific probe set (subcenM-FISH) for chromosome 2, the presence of a small partial trisomy due to a karyotype 47,XX,+r(2)(::p11.1->q11.2::) was demonstrated. Including this case, a total of 11 patients with sSMC(2) are described throughout the literature. Based on that data, a first genotype/phenotype correlation according to the size and structure of the marker is suggested.

Chromosomes, Human, Pair 2↗