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1.
Phys Rev Lett ; 102(13): 132501, 2009 Apr 03.
Article in English | MEDLINE | ID: mdl-19392349

ABSTRACT

High-precision Penning-trap mass measurements of the N approximately Z approximately 34 nuclides 68Se, 70Se, (70m)Br, and 71Br were performed, reaching experimental uncertainties of 0.5-15 keV. The new and improved mass data together with theoretical Coulomb displacement energies were used as input for rp process network calculations. An increase in the effective lifetime of the waiting point nucleus 68Se was found, and more precise information was obtained on the luminosity during a type I x-ray burst along with the final elemental abundances after the burst.

2.
Phys Rev Lett ; 100(13): 132501, 2008 Apr 04.
Article in English | MEDLINE | ID: mdl-18517939

ABSTRACT

A new long-lived isomeric state in (65)Fe has been discovered with Penning trap mass spectrometry and high-precision mass measurements of the neutron-rich isotopes (63-65)Fe and (64-66)Co have been performed with the Low-Energy Beam and Ion Trap Facility at the NSCL. For the new isomer in (65)Fe an excitation energy of 402(5) keV has been determined from the measured mass difference between the isomeric and ground states. The mass uncertainties of all isotopes have been reduced by a factor of 10-100 compared to previous results. In the case of (64)Co the previous mass value was found to deviate by about 5 standard deviations from the new measurement.

3.
Phys Rev Lett ; 96(15): 152501, 2006 Apr 21.
Article in English | MEDLINE | ID: mdl-16712152

ABSTRACT

The mass of the short-lived radio nuclide 38Ca (T(1/2) = 440 ms) has been measured with the 9.4-T Penning trap mass spectrometer of the Low-Energy Beam and Ion Trap Facility. A mass uncertainty of deltam = 280 eV has been achieved, corresponding to deltam/m = 8 x 10(-9). The result makes 38Ca, a superallowed beta emitter, a new candidate to test the conserved-vector-current hypothesis. The experiment is also the first demonstration that short-lived radioactive isotopes produced by projectile fragmentation of relativistic heavy-ion beams can be slowed down and prepared such that precision experiments of this kind are possible.

4.
Brain Res Mol Brain Res ; 96(1-2): 30-8, 2001 Nov 30.
Article in English | MEDLINE | ID: mdl-11731006

ABSTRACT

Apoptosis may represent a prominent form of neuronal death in chronic neurodegenerative disorders, such as Alzheimer's disease. Although apoptosis under mitochondrial control has received considerable attention, mechanisms used within the endoplasmic reticulum (ER) and nucleus in mediating apoptotic signals are not well understood. A growing body of evidence is emerging from different studies which suggests an active role for the ER in regulating apoptosis. Disturbances of ER function have been shown to trigger two different apoptotic pathways; one involves cross-talk with mitochondria and is regulated by the antiapoptotic Bcl-2, and the second is characterized by the activation of caspase-12. Also, stress in the ER has been suggested to result in the activation of a number of proteins, such as gadd 153 and NF-kappa, and in the downregulation of the antiapoptotic protein, Bcl-2. In the present study, the intracisternal injection in aged rabbits of either the neurotoxin aluminum maltolate or of Abeta(1-42), has been found to induce nuclear translocation of gadd 153 and the inducible transcription factor, NF-kappaB. Translocation of these two proteins is accompanied by decreased levels of Bcl-2 in both the ER and the nucleus. Aluminum maltolate, but not Abeta, induces caspase-12 activation which is a mediator of ER-specific apoptosis; this is the first report of the in vivo activation of caspase-12. These findings indicate that the ER may play a role in regulating apoptosis in vivo, and could be of significance in the pathology of neurodegeneration and related disorders.


Subject(s)
Amyloid beta-Peptides/toxicity , CCAAT-Enhancer-Binding Proteins/metabolism , Endoplasmic Reticulum/metabolism , Hippocampus/pathology , NF-kappa B/metabolism , Organometallic Compounds/toxicity , Peptide Fragments/toxicity , Pyrones/toxicity , Transcription Factors/metabolism , Active Transport, Cell Nucleus/physiology , Animals , Apoptosis/drug effects , Blotting, Western , CCAAT-Enhancer-Binding Proteins/analysis , Caspase 12 , Caspases/analysis , Caspases/metabolism , Hippocampus/metabolism , Immunohistochemistry , Male , NF-kappa B/analysis , Nerve Degeneration/chemically induced , Nerve Degeneration/metabolism , Nerve Degeneration/pathology , Proto-Oncogene Proteins c-bcl-2/analysis , Proto-Oncogene Proteins c-bcl-2/metabolism , Rabbits , Transcription Factor CHOP , Transcription Factors/analysis
6.
Neurobiol Dis ; 8(5): 764-73, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11592846

ABSTRACT

Direct (intracisternal) injection of aluminum complexes into rabbit brain results in a number of similarities with the neuropathological and biochemical changes observed in Alzheimer's disease and provides the opportunity to assess early events in neurodegeneration. This mode of administration induces cytochrome c release from mitochondria, a decrease in Bcl-2 in both mitochondria and endoplasmic reticulum, Bax translocation into mitochondria, activation of caspase-3, and DNA fragmentation. Coadministration of glial cell neuronal-derived factor (GDNF) inhibits these Bcl-2 and Bax changes, upregulates Bcl-XL, and abolishes the caspase-3 activity. Furthermore, treatment with GDNF dramatically inhibits apoptosis, as assessed by the TUNEL technique for detecting DNA damage. Treatment with GDNF may represent a therapeutic strategy to reverse the neuronal death associated with Alzheimer's disease and may exert its effect on apoptosis-regulatory proteins.


Subject(s)
Apoptosis/drug effects , Hippocampus/drug effects , Mitochondria/metabolism , Nerve Growth Factors , Nerve Tissue Proteins/biosynthesis , Nerve Tissue Proteins/therapeutic use , Neurons/drug effects , Proto-Oncogene Proteins c-bcl-2/biosynthesis , Proto-Oncogene Proteins/metabolism , Alzheimer Disease/drug therapy , Alzheimer Disease/metabolism , Animals , Caspase 3 , Caspases/metabolism , Cisterna Magna , Cytochrome c Group/metabolism , DNA Fragmentation , Drug Evaluation, Preclinical , Endoplasmic Reticulum/metabolism , Female , Gene Expression Regulation/drug effects , Genes, bcl-2 , Glial Cell Line-Derived Neurotrophic Factor , Hippocampus/metabolism , Hippocampus/pathology , In Situ Nick-End Labeling , Injections , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/pharmacology , Neurons/metabolism , Neurons/pathology , Organometallic Compounds/administration & dosage , Organometallic Compounds/toxicity , Protein Transport/drug effects , Proto-Oncogene Proteins c-bcl-2/genetics , Pyrones/administration & dosage , Pyrones/toxicity , Rabbits , bcl-2-Associated X Protein , bcl-X Protein
7.
Brain Res ; 903(1-2): 66-73, 2001 Jun 08.
Article in English | MEDLINE | ID: mdl-11382389

ABSTRACT

Neurodegenerative diseases, including Alzheimer's disease, are characterized by a progressive and selective loss of neurons. Apoptosis under mitochondrial control has been implicated in this neuronal death process, involving the release of cytochrome c into the cytoplasm and initiation of the apoptosis cascade. However, a growing body of evidence suggests an active role for the endoplasmic reticulum in regulating apoptosis, either independent of mitochondrial, or in concert with mitochondrial-initiated pathways. Members of the Bcl-2 family of proteins have been shown to either inhibit apoptosis, as is the case with Bcl-2, or to promote it, in the case of Bax. Investigations in our laboratory have focused on neuronal injury resulting from the intracisternal administration of aluminum maltolate to New Zealand white rabbits, an animal system relevant to a study of human disease in that it reflects many of the histological and biochemical changes associated with Alzheimer's disease. Here we report that treatment of young adult rabbits with aluminum maltolate induces both cytochrome c translocation into brain cytosol, and caspase-3 activation. Furthermore, as assessed by Western blot analysis, these effects are accompanied by a decrease in Bcl-2 and an increase in Bax reactivity in the endoplasmic reticulum.


Subject(s)
Alzheimer Disease/metabolism , Apoptosis/physiology , Endoplasmic Reticulum/metabolism , Mitochondria/metabolism , Neurons/cytology , Aluminum , Alzheimer Disease/chemically induced , Alzheimer Disease/pathology , Animals , Blotting, Western , Caspase 3 , Caspases/metabolism , Cytochrome c Group/analysis , Cytochrome c Group/metabolism , Disease Models, Animal , Female , Hippocampus/cytology , Hippocampus/drug effects , Immunohistochemistry , Neurons/chemistry , Neurons/enzymology , Proto-Oncogene Proteins/analysis , Proto-Oncogene Proteins/metabolism , Proto-Oncogene Proteins c-bcl-2/analysis , Proto-Oncogene Proteins c-bcl-2/metabolism , Rabbits , Subcellular Fractions , bcl-2-Associated X Protein
8.
Clin Chem ; 47(3): 418-25, 2001 Mar.
Article in English | MEDLINE | ID: mdl-11238291

ABSTRACT

BACKGROUND: C-Reactive protein (CRP) can provide prognostic information about risk of future coronary events in apparently healthy subjects. This application requires higher sensitivity assays than have traditionally been available in the clinical laboratory. METHODS: Nine high-sensitivity CRP (hs-CRP) methods from Dade Behring, Daiichi, Denka Seiken, Diagnostic Products Corporation, Iatron, Kamiya, Olympus, Roche, and Wako were evaluated for limit of detection, linearity, precision, prozone effect, and comparability with samples from 388 apparently healthy individuals. RESULTS: All methods had limits of detection that were lower than the manufacturers' claimed limit of quantification except for the Kamiya, Roche, and Wako methods. All methods were linear at 0.3-10 mg/L. The Diagnostic Products Corporation, Kamiya, Olympus, and Wako methods had imprecision (CVs) >10% at 0.15 mg/L. The Iatron, Olympus, and Wako methods demonstrated prozone effects at hs-CRP concentrations of 12, 206, and 117 mg/L, respectively. hs-CRP concentrations demarcating each quartile in a healthy population were method-dependent. Ninety-two to 95% of subjects were classified into the same quartile of hs-CRP established by the Dade Behring method by the Denka Seiken, Diagnostic Products Corporation, Iatron, and Wako methods. In contrast, 68-77% of subjects were classified into the same quartile by the Daiichi, Kamiya, Olympus, and Roche methods. No subject varied by more than one quartile by any method. CONCLUSIONS: Four of the nine examined hs-CRP methods classified apparently healthy subjects into quartiles of hs-CRP similar to the classifications assigned by the comparison method. Additional standardization efforts are required because an individual patient's results will be interpreted using population-based cutpoints.


Subject(s)
C-Reactive Protein/analysis , Adolescent , Adult , Aged , Autoanalysis , Blood Donors , Female , Humans , Immunoassay , Luminescent Measurements , Male , Middle Aged , Nephelometry and Turbidimetry , Reagent Kits, Diagnostic , Regression Analysis
9.
Clin Chem ; 47(1): 118-23, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11148186

ABSTRACT

BACKGROUND: Staffing core laboratories with appropriate skilled workers requires a process to schedule these individuals so that all workstations are appropriately filled and all the skills of each worker are exercised periodically to maintain competence. METHODS: We applied a genetic algorithm to scheduling laboratory personnel. Our program, developed in Visual Basic 4.0, maximizes the value of a fitness function that measures how well a given scheduling of individuals and their skills matches a set of work tasks for a given work shift. The user provides in an Excel spreadsheet the work tasks, individuals available to work on any given date, and skills each individual possesses. The user also specifies the work shift to be scheduled, the range of dates to be scheduled, the number of days that an individual stays on a given workstation before rotating, and various parameters for the genetic algorithm if they differ from the default values. RESULTS: For >22 months, the program matched individuals to those tasks for which they were qualified and maintained personnel skills by rotating job duties. The schedules generated by the program allowed supervisory personnel to anticipate dates far in advance of when worker availability would be limited, so staffing could be adjusted. In addition, the program helped to identify skills for which too few individuals had been trained. This program has been well accepted by the staff in the clinical laboratories of a 670-bed university medical center, saving 37 h of labor per month, or approximately $11,000 per year, in time that supervisory personnel have spent developing work schedules. CONCLUSIONS: The genetic algorithm approach appears to be useful for scheduling in highly technical work environments that employ multiskilled workers.


Subject(s)
Algorithms , Laboratories , Personnel Staffing and Scheduling , Computers , Hospital Bed Capacity, 500 and over , Hospitals, University , Models, Theoretical , Workforce
10.
Mol Cell Biol ; 21(3): 781-93, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11154266

ABSTRACT

Steroid hormone receptors act to regulate specific gene transcription primarily as steroid-specific dimers bound to palindromic DNA response elements. DNA-dependent dimerization contacts mediated between the receptor DNA binding domains stabilize DNA binding. Additionally, some steroid receptors dimerize prior to their arrival on DNA through interactions mediated through the receptor ligand binding domain. In this report, we describe the steroid-induced homomeric interaction of the rat glucocorticoid receptor (GR) in solution in vivo. Our results demonstrate that GR interacts in solution at least as a dimer, and we have delimited this interaction to a novel interface within the hinge region of GR that appears to be both necessary and sufficient for direct binding. Strikingly, we also demonstrate an interaction between GR and the mineralocorticoid receptor in solution in vivo that is dependent on the ligand binding domain of GR alone and is separable from homodimerization of the glucocorticoid receptor. These results indicate that functional interactions between the glucocorticoid and mineralocorticoid receptors in activating specific gene transcription are probably more complex than has been previously appreciated.


Subject(s)
Receptors, Glucocorticoid/chemistry , Receptors, Mineralocorticoid/chemistry , Animals , Binding Sites , COS Cells , Cell Line , Cell Nucleus/metabolism , Cytoplasm , Dimerization , In Vitro Techniques , Protein Structure, Quaternary , Rats , Receptors, Glucocorticoid/genetics , Receptors, Glucocorticoid/metabolism , Receptors, Mineralocorticoid/genetics , Receptors, Mineralocorticoid/metabolism , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Solutions , Two-Hybrid System Techniques
11.
Neurotoxicology ; 20(5): 805-17, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10591516

ABSTRACT

We propose that aging is an important factor in the susceptibility of neurons to oxidative stress and to subsequent apoptosis. In the present report we demonstrate that aged rabbits treated intracisternally with aluminum maltolate exhibit intense intraneuronal silver positivity indicative of the formation of neurofilamentous aggregates, together with oxidative stress. These changes occur in the CA1 region of the hippocampus as well as in cerebral cortical areas. Apoptosis, measured by the TUNEL in situ technique, colocalizes with oxidative stress. Young animals treated with aluminum show few of these alterations, while age-matched controls are essentially negative. Further studies on the time course of these and related changes demonstrate that oxidative stress and redox-active iron accumulation in hippocampal neurons occur very rapidly, within a period of 3 hours, and increased in intensity at 72 hours. Changes suggestive of apoptosis are seen by 24 hours and are pronounced at 72 hours. In aged animals there is an initially intense immunopositivity at 3 hours for Bcl-2, with negative staining for Bax. By 72 hours, when apoptosis is strongly evident, Bcl-2 is negative and Bax strongly positive. In contrast to the aged rabbits, young animals treated similarly with aluminum exhibit much less oxidative stress with no apoptosis, and maintain Bcl-2 immunopositivity and negative Bax staining. Our findings strongly support the key role that oxidative damage plays in the process of neurodegeneration and in the increased vulnerability to aluminum-induced injury in the aged animal. These are novel observations which may have important implications for aiding in our understanding of the pathogenesis of neurodegeneration occurring in Alzheimer's disease.


Subject(s)
Aging/physiology , Aluminum/toxicity , Apoptosis/drug effects , Genes, bcl-2/drug effects , Hippocampus/growth & development , Iron/metabolism , Nerve Degeneration/chemically induced , Nerve Degeneration/pathology , Oxidative Stress/drug effects , Proto-Oncogene Proteins c-bcl-2 , Proto-Oncogene Proteins/genetics , Animals , Antibodies, Monoclonal/metabolism , Female , Gene Expression Regulation/drug effects , Hippocampus/drug effects , Hippocampus/metabolism , Nerve Degeneration/metabolism , Neurofibrillary Tangles/drug effects , Neurofibrillary Tangles/physiology , Neurofilament Proteins/metabolism , Oxidation-Reduction , Rabbits , Silver Staining , bcl-2-Associated X Protein , tau Proteins/metabolism
13.
Ann Clin Lab Sci ; 29(2): 118-26, 1999.
Article in English | MEDLINE | ID: mdl-10219699

ABSTRACT

Progress in understanding the role of trace metals in biology has been largely dependent on the development of sensitive, accurate and precise analytical methods. Atomic spectroscopic techniques, particularly atomic absorption, have made the greatest contribution. Key to the success of such analytical techniques has been the simplification of sample processing so that contamination is minimized. Electrothermal atomization has allowed sensitivity limits to be lowered sufficiently so that even ultra-trace metals can be detected. More recently, mass spectrometric detection of metal ions has added to the repertoire of available instrumentation, particularly with the use of inductively coupled plasma to introduce ions into the mass analyzer. These analyzers are suitable for multielement analysis. More conventional mass spectrometric analysis of metal chelates offer an alternate solution but require considerable specimen preparation time. Intracellular localization of trace metals necessitates complex specimen processing prior to analysis on instruments that are highly sophisticated and expensive. Metal speciation is a rapidly growing area of trace metal research, with the major advances coming from coupling of the separation process, such as capillary electrophoresis or high performance liquid chromatography, with the analytical instrument for metal detection. Inductively coupled plasma-mass spectrometry has proved to be an excellent choice for such detection purposes. Refinement of these methods as well as more widely available instruments for microanalysis will add greatly to continued advances in our knowledge of the role of trace metals in biology and medicine.


Subject(s)
Metals/analysis , Trace Elements/analysis , Animals , Chemistry Techniques, Analytical/methods , Humans , Lasers , Mass Spectrometry , Methods , Robotics
14.
Mol Cell Biol ; 19(2): 1025-37, 1999 Feb.
Article in English | MEDLINE | ID: mdl-9891038

ABSTRACT

Glucocorticoid receptor (GR) cycles between a free liganded form that is localized to the nucleus and a heat shock protein (hsp)-immunophilin-complexed, unliganded form that is usually localized to the cytoplasm but that can also be nuclear. In addition, rapid nucleocytoplasmic exchange or shuttling of the receptor underlies its localization. Nuclear import of liganded GR is mediated through a well-characterized sequence, NL1, adjacent to the receptor DNA binding domain and a second, uncharacterized motif, NL2, that overlaps with the ligand binding domain. In this study we report that rapid nuclear import (half-life [t1/2] of 4 to 6 min) of agonist- and antagonist-treated GR and the localization of unliganded, hsp-associated GRs to the nucleus in G0 are mediated through NL1 and correlate with the binding of GR to pendulin/importin alpha. By contrast, NL2-mediated nuclear transfer of GR occurred more slowly (t1/2 = 45 min to 1 h), was agonist specific, and appeared to be independent of binding to importin alpha. Together, these results suggest that NL2 mediates the nuclear import of GR through an alternative nuclear import pathway. Nuclear export of GR was inhibited by leptomycin B, suggesting that the transfer of GR to the cytoplasm is mediated through the CRM1-dependent pathway. Inhibition of GR nuclear export by leptomycin B enhanced the nuclear localization of both unliganded, wild-type GR and hormone-treated NL1(-) GR. These results highlight that the subcellular localization of both liganded and unliganded GRs is determined, at least in part, by a flexible equilibrium between the rates of nuclear import and export.


Subject(s)
Receptors, Glucocorticoid/metabolism , Amino Acid Sequence , Amino Acid Substitution , Animals , Binding Sites/genetics , Biological Transport, Active , COS Cells , Cell Nucleus , Chloramphenicol O-Acetyltransferase/genetics , Cytoplasm/metabolism , Genes, Reporter , Ligands , Molecular Sequence Data , Mutagenesis, Site-Directed , Nuclear Proteins/metabolism , Receptors, Glucocorticoid/chemistry , Receptors, Glucocorticoid/genetics , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Saccharomyces cerevisiae/genetics , Transcriptional Activation , Transfection
15.
J Biol Chem ; 274(3): 1432-9, 1999 Jan 15.
Article in English | MEDLINE | ID: mdl-9880517

ABSTRACT

Glucocorticoid receptor (GR) recycles between an inactive form complexed with heat shock proteins (hsps) and localized to the cytoplasm and a free liganded form that regulates specific gene transcription in the nucleus. We report here that, contrary to previous assumptions, association of GR into hsp-containing complexes is not sufficient to prevent the shuttling or trafficking of the GR across the nuclear membrane. Following the withdrawal of treatment with cortisol or the hormone antagonist RU486, GRs recycled rapidly into hsp-associated, hormone-responsive complexes. However, cortisol-withdrawn receptors redistributed to the cytoplasm very slowly (t(1)/(2) = 8-9 h) and RU486-withdrawn receptors not at all. Persistent localization of these GRs to the nucleus was not due to a gross defect in export, since in both instances the complexed nuclear GRs transferred efficiently between heterokaryon nuclei. Moreover, the addition of a nuclear retention signal to the N terminus of GR induced the transfer of naive receptor to the nucleus in the absence of steroid. These results suggest that the localization of GR to the cytoplasm is determined by fine control of the rates of transfer of GR across the nuclear membrane and/or by active retention that occurs independently from the association of GR with hsps.


Subject(s)
Cell Nucleus/metabolism , Cytoplasm/metabolism , Heat-Shock Proteins/metabolism , Receptors, Glucocorticoid/metabolism , 3T3 Cells , Animals , Biological Transport , Bisbenzimidazole/metabolism , COS Cells , Fluorescent Dyes/metabolism , Hormone Antagonists/pharmacology , Hydrocortisone/pharmacology , Ligands , Mice , Mifepristone/pharmacology , Protein Conformation , Rats , Receptors, Glucocorticoid/drug effects , Structure-Activity Relationship , Transcription, Genetic , Tumor Cells, Cultured
17.
Mol Endocrinol ; 12(11): 1749-63, 1998 Nov.
Article in English | MEDLINE | ID: mdl-9817600

ABSTRACT

We report glucocorticoid-dependent induction of transcription from the herpes simplex virus thymidine kinase gene promoter proximal regulatory region in the absence of glucocorticoid response elements and independent of the ability of glucocorticoid receptor (GR) to bind DNA. Examination of the thymidine kinase promoter localized glucocorticoid responsiveness to a binding site for CCAAT enhancer-binding proteins (C/EBPs). Further analysis indicated that GR specifically potentiated the induction of transcription by C/EBP beta, but not C/EBP alpha or delta, and that full induction could be obtained by the ligand-binding domain (LBD) of GR alone. C/EBP beta, but not C/EBP alpha or delta, reciprocally potentiated transcriptional activation by DNA-bound GR LBD. However, C/EBP beta was unable to increase activation by a GR LBD with a short C-terminal truncation, indicating that the functional interaction between the two factors was dependent upon the GR AF-2. Surprisingly, despite the specificity in functional effects, all three C/EBPs bound indistinguishably to GR in GST pull-down and immunoprecipitation assays. Indeed, several nuclear receptors, including the estrogen (ER alpha), progesterone, retinoic acid (RAR), and androgen receptors, displayed a similar potential to bind C/EBPs. Previous reports have demonstrated that ER alpha and RARs repress transcriptional activation by C/EBP beta in ways that were dependent on their related AF-2 functions. Therefore, the GR AF-2 may encode functional features that distinguish the transcriptional regulatory potential of GR from that of ER and RAR. Finally, C/EBP binding mapped to the GR DNA-binding domain, which was not required for functional interaction with C/EBP beta. Thus, the potentiation of C/EBP beta-mediated transcription by GR would appear to require the presence of an intermediary factor.


Subject(s)
DNA-Binding Proteins/physiology , Enhancer Elements, Genetic , Nuclear Proteins/physiology , Protein Isoforms/chemistry , Protein Structure, Tertiary , Receptors, Glucocorticoid/chemistry , Transcriptional Activation/drug effects , Animals , CCAAT-Enhancer-Binding Proteins , COS Cells , Genes, Viral , Promoter Regions, Genetic , Protein Binding , Protein Isoforms/physiology , Rats , Receptors, Glucocorticoid/physiology , Receptors, Steroid/chemistry , Recombinant Fusion Proteins/genetics , Simplexvirus/genetics , Structure-Activity Relationship , Thymidine Kinase/biosynthesis , Thymidine Kinase/genetics , Transfection , Viral Proteins/biosynthesis , Viral Proteins/genetics
18.
Ann Clin Lab Sci ; 28(3): 131-7, 1998.
Article in English | MEDLINE | ID: mdl-9646852

ABSTRACT

The in situ detection of cells undergoing apoptosis is increasingly important in the analysis of injury and degeneration in the central nervous system. Limited information is presently available on the quantification of apoptosis in paraffin-embedded brain tissue sections, a technique which would be most useful in the evaluation of archival tissue for diagnostic and experimental purposes. In this report, optimized conditions for tissue digestion and permeabilization using Proteinase K and Triton X and a quantification method for apoptosis detection are described using brain sections from aluminum maltolate-treated aged and young rabbits as compared to untreated matched controls. This method provides optimal staining of apoptotic cells without the problem of tissue destruction, and should prove useful in evaluating the process of apoptosis in neurodegenerative disorders.


Subject(s)
Apoptosis , Brain/pathology , Neurodegenerative Diseases/pathology , Paraffin , Tissue Embedding , Aging , Animals , DNA Fragmentation , DNA Nucleotidylexotransferase , Deoxyuracil Nucleotides , Endopeptidase K , Female , Hippocampus/pathology , Neurodegenerative Diseases/chemically induced , Organometallic Compounds , Pyrones , Rabbits
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