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At least 289 records · Page 16Linked to original sources
PubMed2001

The use of multicolor fluorescence technologies in the characterization of prostate carcinoma cell lines: a comparison of multiplex fluorescence in situ hybridization and spectral karyotyping data.

Recent studies have identified several chromosome regions that are altered in primary prostate cancer and prostatic carcinoma cell lines. These targeted regions may harbor genes involved in tumor suppression. We used multiplex fluorescence in situ hybridization (M-FISH) to screen for genetic rearrangements in four prostate cancer cell lines, LNCaP, LNCaP.FCG, DU145, and PC3, and compared our results with those recently obtained using spectral karyotyping (SKY). A number of differences was noted between abnormalities characterized by SKY and M-FISH, suggesting variation in karyotype evolution and characterization by these two methodologies. M-FISH analysis showed that hormone-resistant cell lines (DU145 and PC3) contained many genetic alterations (> or =15 per cell), suggesting high levels of genetic instability in hormone-refractory prostate cancer. Most chromosome regions previously implicated in prostate cancer were altered in one or more of these cell lines. Several specific chromosome aberrations were also detected, including a del(4)(p14) and a del(6)(q21) in the hormone-insensitive cell lines, a t(1;15)(p?;q?) in LNCaP, LNCaP, and PC3, and a i(5p) in LNCaP.FCG, DU145, and PC3. These clonal chromosome abnormalities may pinpoint gene loci associated with prostate tumourigenesis, cancer progression, and hormone sensitivity.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1998

Hidden chromosome abnormalities in a primary central nervous system lymphoma detected by multicolor spectral karyotyping.

Cytogenetic analysis provides important information for diagnosis and prognosis in some tumors. But karyotype analysis can be difficult in some cases, because metaphase chromosomes are contracted. New techniques, such as fluorescence in situ hybridization and, more recently, spectral karyotyping, or SKY, based on the hybridization of 24 fluorescently labeled chromosome painting probes, allow the detection and identification of complex chromosomal rearrangements. We report here a case of primary central nervous system lymphoma in which chromosomal rearrangements and marker chromosomes not identified by a routine cytogenetic technique were clarified by SKY. This shows the value of the SKY technique in the cytogenetic diagnosis of tumors.

Aged↗
PubMed1999

Coexistence of several unbalanced translocations in a case of neuroblastoma: the contribution of multicolor spectral karyotyping.

Spectral karyotyping (SKY) is based on the simultaneous hybridization of a set of 24 chromosome-specific DNA painting probes, each labeled with a different fluor combination. Automatic classification, based on the measurement of the spectrum for each chromosome, was applied to metaphases obtained from the affected bone marrow of a neuroblastoma case. Spectral karyotyping allowed the identification of chromosomal aberrations that could not be identified by the use of the G-banding technique, and revealed a number of gains and unbalanced translocations.

Child, Preschool↗
PubMed2000

Enhanced detection of chromosomal abnormalities with the use of RxFISH multicolor banding technique.

Hematological disorders often have complex karyotypes with multiple markers. Proper assignment of chromosome number or aberration or both can be difficult. Specific identification of chromosomal abnormalities aids in the diagnosis and selection of treatment of patients. Fluorescence in situ hybridization (FISH) has been applied to the identification of translocations, markers, and other chromosomal abnormalities in clinical cytogenetics. However, the standard FISH technique is unable to detect the entire genome in a single experiment. This report presents the use of a cross-species comparative genomic hybridization color-banding technique (RxFISH) that permits examination of an entire karyotype at one time. Specimens from two patients, one with acute lymphocytic leukemia (ALL) and the other with multiple myeloma (MM), were studied. Metaphases were prepared by standard culture techniques. Conventional cytogenetic analysis (GTG banding) showed multiple clones in each of the cases. These clones were hyperdiploid metaphases with complex chromosomal abnormalities and multiple markers. The slides were then hybridized with FITC-, Cy-3-, and Cy-5-labeled RxFISH probes; the results were analyzed by a digital imaging system. The RxFISH color banding confirmed the hyperdiploid metaphases and identified multiple chromosomal abnormalities. In the specimen from the patient with ALL, several chromosomes, which had been classified as markers by G-banding, were found to be specific chromosomes. This study suggests that RxFISH can provide more accurate and specific identification of complex chromosomal abnormalities. RxFISH is a useful complement to the clinical cytogenetic laboratory armamentarium.

Bone Marrow↗
PubMed2000

Multicolor "DiOlistic" labeling of the nervous system using lipophilic dye combinations.

We describe a technique for rapid labeling of a large number of cells in the nervous system with many different colors. By delivering lipophilic dye-coated particles to neuronal preparations with a "gene gun," individual neurons and glia whose membranes are contacted by the particles are quickly labeled. Using particles that are each coated with different combinations of various lipophilic dyes, many cells within a complex neuronal network can be simultaneously labeled with a wide variety of colors. This approach is most effective in living material but also labels previously fixed material. In living material, labeled neurons continue to show normal synaptic responses and undergo dendritic remodeling. This technique is thus useful for studying structural plasticity of neuronal circuits in living preparations. In addition, the Golgi-like labeling of neurons with many different colors provides a novel way to study neuronal connectivity.

Animals↗
PubMed2005

Detection of medically important Ehrlichia by quantitative multicolor TaqMan real-time polymerase chain reaction of the dsb gene.

Ehrlichia species are the etiological agents of emerging and life-threatening tick-borne human zoonoses, in addition to causing serious and fatal infections in companion animals and livestock. We developed the first tricolor TaqMan real-time polymerase chain reaction assay capable of simultaneously detecting and discriminating medically important ehrlichiae in a single reaction. Analytical sensitivity of 50 copies per reaction was attained with templates from Ehrlichia chaffeensis, Ehrlichia ewingii, and Ehrlichia canis by amplifying the genus-specific disulfide bond formation protein gene (dsb). Ehrlichia genus-specific dsb primers amplified DNA from all known Ehrlichia species but not from other rickettsial organisms including Anaplasma platys, Anaplasma phagocytophilum, Rickettsia conorii, or Rickettsia typhi. High species specificity was attained as each species-specific TaqMan probe (E. chaffeensis, E. ewingii, and E. canis) identified homologous templates but did not cross-hybridize with heterologous Ehrlichia templates at concentrations as high as 10(8) copies. Identification of E. chaffeensis, E. ewingii, and E. canis from natural and experimental infections, previously confirmed by polymerase chain reaction and serological or microscopic evidence, demonstrated the comparable specificity and sensitivity of the dsb real-time assay. This assay provides a powerful tool for prospective medical diagnosis for human and canine ehrlichioses and for ecologic and epidemiological studies involving arthropod and mammalian hosts.

Animals↗
PubMed2004

Single color fluorescent indicators of protein phosphorylation for multicolor imaging of intracellular signal flow dynamics.

Existing monitoring methods for protein phosphorylation involved in intracellular signal transduction in vivo are exclusively based on fluorescence resonance energy transfer, which needs the measurement of the change in fluorescence intensities at two wavelengths. Therefore, it is difficult to monitor protein phosphorylation together with other related signaling processes, such as second messengers and protein translocation. To overcome this problem, we developed novel fluorescent indicators, each containing a differently colored (cyan and green) single fluorophore. The present indicator is a tandem fusion protein containing a kinase substrate domain, a circularly permuted fluorescent protein (cpFP), and a phosphorylation recognition domain. The cpFP is obtained by dividing a green fluorescent protein mutant (GFP) at residue 144-145 and linking the carboxy and amino portions thereof with a peptide linker. The substrate domain used in this study is a peptide sequence that is phosphorylated by insulin receptor. Phosphorylation of the substrate domain induces its interaction with the phosphorylation recognition domain, which causes a conformational change in the cpFP and a change in its fluorescence. The cyan and green indicators exhibited 10% decrease and 15% increase, respectively, in their fluorescence intensities upon phosphorylation. Using this cyan indicator and GFP-tagged mitogen-activated protein kinase (MAPK), we found that insulin-induced protein phosphorylation occurred immediately upon the addition of insulin, whereas nuclear translocation of MAPK occurred 7 min later. By tailoring the substrate domains and the phosphorylation recognition domains in these cyan and green indicators, the present approach should be applicable to the in vivo analysis of a broad range of protein phosphorylation processes, together with other intracellular signaling processes.

Amino Acid Sequence↗
PubMed2006

Polychromatic microarrays: simultaneous multicolor array hybridization of eight samples.

High-throughput microscale platforms have transformed modern analytical investigations. Traditional microarray analyses involve a comparative approach, with two samples, a known control and an unknown sample, hybridized side-by-side and then contrasted for genetic differences. The samples are labeled with separate dyes and hybridized together, providing a differential expression pattern based on the reporter intensities. In contrast, the fiber-optic microarray platform described herein is analyzed with a microscope, thereby enabling the use of virtually any reporter, including quantum dots. The instrumentation takes advantage of the narrow emission bands characteristic of quantum dots to perform multiplexed detection of Bacillus anthracis. Advancing beyond the standard red/green microarray experiment, a panel of eight reporters were linked to eight B. anthracis samples and simultaneously analyzed in a microarray format. The ability to employ an assortment of reporters, along with the capacity to simultaneously hybridize eight samples confers an unprecedented flexibility to array-based analyses, providing a 4-fold increase in throughput over standard two-color assays.

Bacillus anthracis↗