Abstract: Aspects of the invention concern methods for detecting, identifying and evaluating tobacco and tobacco products to determine the potential that these compositions have to contribute to a tobacco-related disease. It is based, at least in part; on the discovery that exposure of pulmonary cells to smoke or smoke condensate obtained from tobacco or tobacco products induces double stranded breaks in cellular DNA, which were efficiently detected using assays that measure the presence, absence, or amount of phosphorylation of the histone, H2AX.

Abstract: Aspects of the invention concern methods for detecting, identifying and evaluating tobacco and tobacco products to determine the potential that these compositions have to contribute to a tobacco-related disease. It is based, at least in part; on the discovery that exposure of pulmonary cells to smoke or smoke condensate obtained from tobacco or tobacco products induces double stranded breaks in cellular DNA, which were efficiently detected using assays that measure the presence, absence, or amount of phosphorylation of the histone, H2AX.

Abstract: Aspects of the invention concern methods for detecting, identifying and evaluating tobacco and tobacco products to determine the potential that these compositions have to contribute to a tobacco-related disease. It is based, at least in part; on the discovery that exposure of pulmonary cells to smoke or smoke condensate obtained from tobacco or tobacco products induces double stranded breaks in cellular DNA, which were efficiently detected using assays that measure the presence, absence, or amount of phosphorylation of the histone, H2AX.

Abstract: Aspects of the invention concern methods for detecting, identifying and evaluating tobacco and tobacco products to determine the potential that these compositions have to contribute to a tobacco-related disease. It is based, at least in part; on the discovery that exposure of pulmonary cells to smoke or smoke condensate obtained from tobacco or tobacco products induces double stranded breaks in cellular DNA, which were efficiently detected using assays that measure the presence, absence, or amount of phosphorylation of the histone, H2AX.

Abstract: The invention provides novel affinity labels for Ser proteases of formula: L-A-X—NH—CH(R?)C(?O)CH2Cl??(I) wherein L, A, X, and R? have any of the values defined in the specification, or salts thereof, as well as compositions comprising such compounds or salts. The composition of the amino acid side-chain (R?) along with the amino acid or amino acid sequence (peptide) of the X component of formula I, affect the target selectivity of the labeled affinity ligand. Utilization of cell permeable, enzyme selective, labeled affinity ligands, provides a precise mechanism for evaluating the current and future status of cell populations.

Abstract: The invention provides assay methods and reagents useful for evaluating the level of enzyme activities within living cells. Enzyme activity levels within living cells, such as Serine proteases, can be key determinates in assessing; 1) the presence of tumor (cancer) cells, 2) the predictive efficacy of a chemotherapeutic treatment regimen using a particular therapeutic agent or process, 4) the probability of graft rejection or acceptance, and 5) the disease state status of a cell. The identification of the up or down regulation relationships of serine proteases within living cell systems, provides a rapid, yet finely tuned mechanism for predicting the current and future physiological state of these cell populations.

Abstract: Methods, reagents, and kits are provided that permit flow cytometric determination of the phosphorylation status of retinoblastoma susceptibility gene protein (pRB) in individual cells. Methods are described that permit the hypophosphorylated, active, form of pRB to be measured either as an absolute quantity or as a proportion of total cellular pRB. Further described are methods that permit pRB phosphorylation status to be correlated with cell cycle phase and with protein components of the cell cycle. Screening of chemical compounds for antiproliferative and antineoplastic activity using the flow cytometric assays is demonstrated. Reagent kits that facilitate the subject methods are also provided.

Abstract: The invention provides assay methods and reagents useful for evaluating the level of enzyme activities within living cells. Enzyme activity levels within living cells, such as caspases and Serine proteases, can be key determinates in assessing; 1) the apoptotic state of a cell, 2) the presence of tumor (cancer) cells, 3) the predictive efficacy of a chemotherapeutic treatment regimen using a particular therapeutic agent or process, 4) the probability of graft rejection or acceptance, identification of the up or down regulation relationships of serine proteases and caspases within living cell systems, provides a rapid, yet finely tuned mechanism for predicting the current and future state of these cell populations, and 5) the disease state status of a cell.

Abstract: Methods, reagents, and kits are provided that permit flow cytometric determination of the phosphorylation status of retinoblastoma susceptibility gene protein (pRB) in individual cells. Methods are described that permit the hypophosphorylated, active, form of pRB to be measured either as an absolute quantity or as a proportion of total cellular pRB. Further described are methods that permit pRB phosphorylation status to be correlated with cell cycle phase and with protein components of the cell cycle. Screening of chemical compounds for antiproliferative and antineoplastic activity using the flow cytometric assays is demonstrated. Reagent kits that facilitate the subject methods are also provided.

Abstract: Methods, reagents, and kits are provided that permit flow cytometric determination of the phosphorylation status of retinoblastoma susceptibility gene protein (pRB) in individual cells. Methods are described that permit the hypophosphorylated, active, form of pRB to be measured either as an absolute quantity or as a proportion of total cellular pRB. Further described are methods that permit pRB phosphorylation status to be correlated with cell cycle phase and with protein components of the cell cycle. Screening of chemical compounds for antiproliferative and antineoplastic activity using the flow cytometric assays is demonstrated. Reagent kits that facilitate the subject methods are also provided.

Abstract: Methods, reagents, and kits are provided that permit flow cytometric determination of the phosphorylation status of retinoblastoma susceptibility gene protein (pRB) in individual cells. Methods are described that permit the hypophosphorylated, active, form of pRB to be measured either as an absolute quantity or as a proportion of total cellular pRB. Further described are methods that permit pRB phosphorylation status to be correlated with cell cycle phase and with protein components of the cell cycle. Screening of chemical compounds for antiproliferative and antineoplastic activity using the flow cytometric assays is demonstrated. Reagent kits that facilitate the subject methods are also provided.

Abstract: Methods for immunocytochemical detection and isolation of RNA.

Abstract: The invention pertains to the field of DNA detection for basic research, medical diagnostic testing, and forensic testing. Methods are provided for end labeling of DNA strands without a denaturation step so that cellular morphology can be better preserved. The DNA strands are first incubated with a halogenated deoxynucleotide triphosphate, such as brominated deoxyuridine triphosphate (BrdUTP), and an enzyme which can catalyze the addition of the halogenated deoxynucleotide to the 3' OH ends of the DNA strand, such as terminal deoxynucleotidyl transferase (TdT). The resulting modified DNA strands are then incubated with a labeled antibody, such as a fluoresceinated monoclonal antibody, that binds specifically to the halogenated deoxynucleotide. The label is then detected, e.g., by flow cytometry. The methods have utility in detecting apotosis, DNA synthesis and/or repair, and as general methods for end labeling DNA.

Abstract: The primary and/or secondary structure of nucleic acid as well as its content and molecular weight can be analyzed in solutions by light scatter or fluorescence measurements after treatment with 3 or 4 ring aromatic cations which bind to single-stranded nucleic acids by cooperative association and induce their condensation (collapse). Preparative separation of nucleic acids of different types from the mixtures in the solution is also accomplished by the same principle and techniques wherein each condensed acid is removed from solution and the condensation reversed.

Abstract: A process for characterizing sperm motility and viability by staining a sperm sample with Rhodamine 123 and ethidium bromide, and simultaneously measuring the sperm fluorescence emissions at green frequencies 515-575 nm and at red frequencies 600-650 nm, the green counts being correlated with sperm motility and the red counts being correlated with putative dying or dead cells. Additionally, a sample of sperm can be characterized as to type and normality by staining a sample of sperm with acridine orange and simultaneously measuring the sperm fluorescence emissions at green frequencies 515-575 nm and at red frequencies 600-650 nm.