William Geary - Academia.edu (original) (raw)
Papers by William Geary
Laboratory Investigation, Apr 15, 1993
The distribution and binding of phenytoin (PHT) were studied in rat brain using anatomically inta... more The distribution and binding of phenytoin (PHT) were studied in rat brain using anatomically intact tissue. The pattern and kinetics of PHT distribution in vivo were examined with quantitative carbon-i 4 autoradiography. Initially gray matter levels were greater than white matter levels, but after 30 mm the opposite condition was found. At a given time point, the levels of PHT among various gray matter structures as a group or among white matter structures as a group were uniform. The ratio of radio-activity in white matter to that in gray matter was 3:1 1 20 mm after injection of radiolabeled PHT. Thin-layer chromatography showed only PHT in the brain 1 5 mm after injection of PHT and both PHT and its hydroxylated metabolite [5-(p-hydroxyphenyl)-5-phenylhydantoinj 1 20 mm after injection. The proportions of PHT and 5-(p-hydroxyphenyl)-5-phenylhydantoin were the same
Quantitative Neuroanatomy in Transmitter Research, 1985
The goal of this current line of work was to develop quantitative autoradiographic methods to reg... more The goal of this current line of work was to develop quantitative autoradiographic methods to regionally quantify the binding properties (i.e. Kd and Bmax) of opiate ligands in morphine dependent and naive rat brain. Such regional studies were not possible utilizing binding techniques to brain membrane preparations because of the limited availability of tissue from dissection of small brain regions. Even with regional dissection techniques, the fine anatomical resolution available with autoradiography was sacrificed. With quantitative autoradiographic methods, one could ask whether the development of opiate tolerance/dependence occurred as a result of regulation of the affinity or concentration of opiate binding sites in any of one or several regions rich in opiate binding sites.
Blood, 1989
Human 125I-plasminogen bound readily to rat hepatocytes in primary culture at 4 degrees C and at ... more Human 125I-plasminogen bound readily to rat hepatocytes in primary culture at 4 degrees C and at 37 degrees C. Binding was inhibited by lysine and reversed by lysine, epsilon-aminocaproic acid, or nonradiolabeled plasminogen. The Kd for binding of 125I-plasminogen to hepatocytes was 0.59 +/- 0.16 mumol/L, as determined from the saturation isotherm by nonlinear regression (r2 = 0.99) and the Scatchard transformation by linear regression (r2 = 0.93). The number of sites per cell was 14.1 +/- 1.1 x 10(6). Fibrinogen synthesis and secretion by hepatocytes was insufficient to account for the major fraction of plasminogen binding, as determined by enzyme-linked immunosorbent assay (ELISA). Polyacrylamide gel electrophoresis and trichloroacetic acid precipitation studies demonstrated that plasminogen is neither activated nor degraded when bound to hepatocytes at 37 degrees C. Thin slices of whole rat liver (500 microns), isolated and prepared totally at 4 degrees C, bound 125I-plasminogen....
Transplantation Proceedings, 1998
American Journal of Clinical Pathology, 1993
Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 1993
Fifty-three cases of B-cell non-Hodgkin's lymphoma (BNHL) and 41 cases of non-BNHL lesions we... more Fifty-three cases of B-cell non-Hodgkin's lymphoma (BNHL) and 41 cases of non-BNHL lesions were retrospectively evaluated for the quantitative features of restricted surface light chain expression, pan B-cell antigens, pan T-cell antigens, and T-helper and T-suppressor phenotype using flow cytometry. Decision limit analyses were applied to multiple quantitative indices of immunophenotype to establish criteria for the detection of clonal proliferation associated with BNHL or non-BNHL conditions. Two data expressions (percent population and relative ratios) were simultaneously analyzed. The percent population measures were amenable to parametric analyses; the ratio data were amenable to nonparametric analyses only. Acceptable diagnostic specificity and sensitivity were obtained using decision limits of 75% kappa light chain and 65% lambda light chain for the detection of clonal proliferations associated with BNHL. Comparable diagnostic criteria for light chain ratios of 3:1 and 2:...
The Journal of pharmacology and experimental therapeutics, 1983
The binding of a radiolabeled opiate agonist ([3H]etorphine) and antagonist ([3H]naloxone) was st... more The binding of a radiolabeled opiate agonist ([3H]etorphine) and antagonist ([3H]naloxone) was studied using quantitative film autoradiography of rat-brain sections labeled by in vitro dipping methods. The binding activities of both [3H]naloxone and [3H] etorphine were saturable in three brain regions: noncluster striatum, nucleus accumbens and cingulate cortex. Eadie-Hofstee analysis of these regions yielded the following binding affinities and capacities: noncluster striatum binding affinity (KD) +/- S.E. = 1.59 +/- 0.23 nM, maximal binding capacity (Bmax) +/- S.E. = 28.3 +/- 1.9 fmol/mg, S.D. error of the raw data (Erad) = 6.4%; nucleus accumbens, KD +/- S.E. = 1.74 +/- 0.28 nM, Bmax +/- S.E. = 73.3 +/- 5.2 fmol/mg, S.D. (Erad) = 6.2%; cingulate cortex, KD +/- S.E. = 1.44 +/- 0.15 nM, Bmax +/- S.E. = 37.6 +/- 1.4 fmol/mg, S.D. (Erad) = 2.5%. A KD +/- S.E. = 1.72 +/- 0.29 nM, Bmax +/- S.E. = 74.1 +/- 5.3 fmol/mg, S.D. (Erad) = 5.0% was found for [3H]etorphine binding in the nonclu...
The Journal of pharmacology and experimental therapeutics, 1987
The distribution and binding of phenytoin (PHT) were studied in rat brain using anatomically inta... more The distribution and binding of phenytoin (PHT) were studied in rat brain using anatomically intact tissue. The pattern and kinetics of PHT distribution in vivo were examined with quantitative carbon-14 autoradiography. Initially gray matter levels were greater than white matter levels, but after 30 min the opposite condition was found. At a given time point, the levels of PHT among various gray matter structures as a group or among white matter structures as a group were uniform. The ratio of radioactivity in white matter to that in gray matter was 3:1 120 min after injection of radiolabeled PHT. Thin-layer chromatography showed only PHT in the brain 15 min after injection of PHT and both PHT and its hydroxylated metabolite [5-(p-hydroxyphenyl)-5-phenylhydantoin] 120 min after injection. The proportions of PHT and 5-(p-hydroxyphenyl)-5-phenylhydantoin were the same in white as in gray matter. Direct chemical measurements of brain samples obtained after coinjections of tracer amount...
Modern Pathology, 2001
The ability to predict cancer progression may help the clinical management of patients with penil... more The ability to predict cancer progression may help the clinical management of patients with penile squamous cell carcinoma. We studied 22 cases of squamous cell carcinoma of the penis diagnosed between 1989 and 1998. The depth of invasion was measured from the basement membrane of the squamous epithelium to the deepest invasive cancer cells. Cancer progression was defined as the development of lymph node metastasis or distant metastasis. The mean patient age was 63 years and the mean follow-up was 28 months. Ten patients developed cancer progression. The mean depth of invasion among patients with cancer progression was 9.8 mM, as compared to the mean depth of invasion of 4.0 mM among those patients without cancer progression (P ؍ .02). Vascular invasion was also predictive of cancer progression (P ؍ .02). Metastases developed in the majority (6 out of 7) of cases invading more than 6 mM, but developed only in a minority (4 out of 15) of cases invading 6 mM or less. We conclude that depth of invasion and vascular invasion are significant predictors of cancer progression for penile squamous cell carcinoma.
Cell and Tissue Banking, 2001
In vitro proliferation of isolated pancreaticislets has become an area of great interest given th... more In vitro proliferation of isolated pancreaticislets has become an area of great interest given the scarcity of clinicalisletdonors and the islet mass requirements for clinical islet transplantation.Smallintestinal submucosa (SIS), a naturally occurring extracellular matrix, hasbeeninvestigated to promote wound healing, tissue remodeling and cell growth. Thisstudy evaluated recovery and function of isolated canine pancreatic isletsfollowing in vitro tissue culture. Pancreatic islets
Journal of Cutaneous Pathology, 1992
A commercially available antibody to proliferating cell nuclear antigen was used to characterize ... more A commercially available antibody to proliferating cell nuclear antigen was used to characterize and compare proliferating cell populations in paraffin sections of benign, premalignant, and malignant lesions of human epidermis using routine immunohistochemical techniques. Three patterns emerged. An ordered pattern was found in prurigo nodularis and keratoacanthoma, wherein moderately and strongly positive nuclei were distributed in a continuous, basal-suprabasal layer of relatively uniform thickness. There was graded loss and ultimate extinction of PCNA staining in progressively more superficial epidermal cells. A basal dysplastic pattern was found in actinic keratosis and squamous cell carcinoma. Nuclei of essentially all dysplastic cells of both categories expressed PCNA, with a preponderance of strongly positive nuclei. These were localized to basal-suprabasal zones that were often expanded. Loss of PCNA reactivity toward the surface was often abrupt. Bowen's disease exhibited a diffuse dysplastic pattern, wherein large numbers of moderately and strongly positive nuclei, in random array, were present in essentially full thickness distribution. In many fields, however, a layer of cytologically bland basal cells, with faint or no nuclear staining, was interposed between dysplastic epithelium and dermis. This study has demonstrated that proliferating cell populations in epidermal lesions can be assessed with simple, inexpensive methods. There were consistent differences between the proliferating cell populations of the various entities studied, differences that can be reasonably correlated with other known clinical, microscopic, and biologic features of the lesions. This technique should provide an interesting new avenue for study of diverse cutaneous diseases.
![Research paper thumbnail of Regional saturation studies of [3H]naloxone binding in the naive, dependent and withdrawal states](https://attachments.academia-assets.com/87661259/thumbnails/1.jpg)
](Brain Research, 1985
We have examined the saturation features (K d and Bronx) of [3H]naloxone binding in rat brain reg... more We have examined the saturation features (K d and Bronx) of [3H]naloxone binding in rat brain regions in cytoarchitectonically intact tissues of naive, morphine dependent and precipitated withdrawal states using quantitative film autoradiography. Sixty-one saturation experiments in 13 regions of naive rat brains yielded monophasic Eadie-Hofstee plots with a mean (+ S,D.) K a of 1.87 + 0.87 nM and a mean Bma x (+ S.D.) of 101 + 66 fmol/mg. The 61 K d values in naive rats described a normal distribution of regional binding affinities that may reflect the biological variation of a single high affinity binding site. Similar studies in the morphine dependent and prec~p, itated withdrawal states showed no apparent changes in either the K d or Bm~ of regional [aH]naloxone binding. The possibility that in vitro preincubation of tissue sections masked potential alterations in [aH]naloxone binding was examined in competition studies of both morphine and naloxone for 2.0 nM [3H]naloxone binding to striatal sections. Preincubation had no effect on the ability of either opiate agonist or antagonist to compete for [3H]naloxone binding in the naive, morphine dependent or precipitated withdrawal states. The regional data comparing K d and Bma x for [3H]naloxone binding in naive, morphine dependent and precipitated withdrawal states eliminated the possibility that previous efforts to demonstrate alterations in opiate binding were confounded by homogenization and/or limitations of gross dissection. Our data suggest that either potential changes in antagonist binding in dependent and withdrawal states can only be demonstrated under strict in VlVO conditions or that post-binding transduction mechanisms are the sites of adaptive changes in naloxone sensitivity in the states of opiate dependence and withdrawal. * Preliminary reports of these data were given at the 13th Society for Neuroscience Meeting (1983) and the Wenner-Gren Symposium, Stockholm, Sweden (May, 1984).
Brain Research, 1985
The details of quantitative film autoradiography for tritium using tritium plastic standards were... more The details of quantitative film autoradiography for tritium using tritium plastic standards were examined with respect to 3 issues: (1) tritium tissue equivalent (TE) calibration; (2) correction of autoabsorption differences for gray and white matter; and (3) the use of carbon-14 standard sources as a method for quantifying tritium tissue images. Both 3H-tissue and 3H-plastic sources produced linear log-log relationships of 3H-concentration (nCi/mg source weight) vs source optical density (OD) over a reproducible OD range (0.080-0.800). The curves for both 3H-tissue and 3H-plastic were parallel; uncalibrated 3H-plastic standards exhibited a 4-fold higher autoabsorption over 3H-tissue sources for OD values in the linear range. Using chloroform extraction of brains from rats treated with either [14C]deoxyglucose ([IaC]DG) or [3H]deoxyglucose ([3H]DG), we found neither isotope loss nor redistribution after defatting (30% reduction of tissue dry weight). After chloroform extraction, the OD values from both gray and white matter structures containing carbon-14 were unaltered. Gray matter OD values increased by 28.7 + 5.6% (mean + S.D.) in structures containing tritium; white matter structures containing tritium exhibited a 115.9 + 29.3% increase in OD after chloroform extraction. The increase in OD after chloroform extraction was a fixed percent for any given tritium OD value from unextracted tissue when the value was within the linear range of 0.080-0.800 OD units. The magnitude of the higher white matter autoabsorption for tritium was confirmed using tritium impregnated cow brain pastes of variable gray/white mixtures. Chloroform extraction of tissue from [3H]DG treated rats was therefore a suitable procedure for direct correction of regionally heterogeneous autoabsorption of tritium. Finally, the rates of image generation for tritium and carbon-14 sources were compared. The rate of increase of OD with increasing exposure time was found to be equal for 3H-tissue and 3H-plastic images; sources of carbon-14 in plastic, however, exhibited more accelerated rates of image generation when compared to tritium sources (i.e. 3H-and 14C-images did not covary with exposure time). The effect of non-covariance on tritium TE calibrated standards was the overestimation of OD values for 14C-plastic standards with increasing times of exposure (comparison of 4 week images to 1 week images showed errors of 35-40%). Use of carbon-14 sources to quantify tritium-generated images therefore required recalibration of laC-plastic for all exposure times of interest.
Brain Research, 1985
Direct corrections of regional differential tritium autoabsorption in quantitative autoradiograph... more Direct corrections of regional differential tritium autoabsorption in quantitative autoradiography of the central nervous system are made using chloroform extraction of brain sections from rats receiving tritiated 2-deoxyglucose in vivo. Fifty-two regional quenching coefficients varied from 30% in gray structures to 100% in white structures. These data may be used to correct for regional quenching of tritium emissions in autoradiographic studies of rat brain.
Brain Research, 1983
We have studied regional cerebral metabolism by 2-deoxyglucose autoradiography in 67 brain struct... more We have studied regional cerebral metabolism by 2-deoxyglucose autoradiography in 67 brain structures of morphine-dependent rats during fixed dose naloxone precipitated withdrawal. Behavioral indices of withdrawal were studied simultaneously in the same animals used in the cerebral metabolism studies. The effects of cumulative morphine dose (470, 1145 and 2345 mg/kg) with fixed dose (0.5 mg/kg) naloxone precipitated withdrawal upon behavioral and cerebral metabolic measures of the severity of withdrawal were compared. All morphine dependent groups studied exhibited the known behavioral sequelae of naloxone precipitated withdrawal. Qualitative withdrawal signs exhibited by all dependent groups included tachypnea, ptosis, penile erection, ejaculation, diarrhea, urination, salivation, lacrimation, rhinorrhea, and irritability. Quantitative signs of withdrawal for 470, 1145 and 2345 mg/kg cumulative morphine dose groups were as follows (mean __
Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1989
a2-Antiplasmin (a2AP), a serpin proteinase inhibitor with two methionine residues in its reactive... more a2-Antiplasmin (a2AP), a serpin proteinase inhibitor with two methionine residues in its reactive center, was treated with c/s.dicldorodiammineplatinum (||) (c/s-DDP). This compound has been utilized previously to specifically modify methionine residues. After reaction, the a2AP demonstrated decreased inhibitory activity against plasmin, min|plasmin, trypsin and chymotryps|n. The reduction in activity depended on the concentration of ds-DDP; however, the amount of activity retained by the treated a2AP was equivalent with each of the four proteinases, a2AP that was incubated with 1.0 mM c/s-DDP for 3 h at 370C was 90% inactivated. These same conditions resulted |n the binding of only 1.0-1.5 tool of platinum per tool of inhibitor. In experiments with lower concentrations of ds-DDP, the amount of incorporated platinum directly correlated with the amount of inactivated azAP (1:1 stoichiometry). Reactions and functions of azAP that do not result in protelnase inhibition were not affected by c/s-DDP. Cleavage of a2AP by elastase, which occurs near tl*~ pro~eina~e inhibition site, was unaffected. In addition, the affinity of a~AP for the K1-3 region of plasminogen remained unchanged after treatment. These data strongly suggest that the reaction of azAP with ds-DDP involves principally a single site on the inhibitor and that this site is critical for proteinase inhibitory activity. The most likely candidate is the P~ methionine which is adjacent to the peptide bonds cleaved in the proteinase inhibitory reactions but not in the elastase reaction.
Annals of Neurology, 1987
The role of the neuropeptide somatostatin in limbic seizures was studied using electrical stimula... more The role of the neuropeptide somatostatin in limbic seizures was studied using electrical stimulation of the hippocampus in kindled rats. Cysteamine, an agent which selectively and reversibly depletes brain somatostatin stores, had a biphasic action. An early proconvulsant effect was seen within a few hours, consisting of prolonged electrographic seizures in the hippocampus and more severe behavioral convulsions. A later anticonvulsant effect, maximal at 1 to 2 days and dissipating within a week, was manifested by less intense behavioral convulsions without change in the duration of electrical seizure activity. Both effects were dose-dependent. No change in afterdischarge thresholds was detected at any time after the administration of cysteamine. Intraventricular administration of somatostatin to animals with behavioral seizures attenuated by cysteamine treatment restored the responses to precysteamine levels. We conclude that somatostatin facilitates the spread of seizures over limbic circuits from a region of focal seizure initiation.
Neurochemical Research, 1987
Quantitative autoradiography of [14C]deoxyglucose, [14C]iodoantipyrine, and [14C]leucine was used... more Quantitative autoradiography of [14C]deoxyglucose, [14C]iodoantipyrine, and [14C]leucine was used to estimate regional cerebral glucose metabolism, cerebral blood flow, and cerebral protein synthesis, respectively, in rats during morphine dependence and withdrawal. Glucose metabolism was elevated in 19 of 26 selected brain regions; the elevations in glucose metabolism were similar when data were expressed as either optical density ratios or as calculated rate values of mumol/100 gm/min. Restraining the rats produced heterogeneous effects on glucose metabolism during morphine withdrawal (MW). Neither estimated cerebral blood flow nor cerebral protein synthesis were affected by morphine and/or naloxone treatments in either naive or morphine-dependent rats. The data demonstrate that changes in regional cerebral glucose utilization occur independently of blood flow changes and exclude the possibility that regional changes in glucose utilization occur as a consequence of large regional changes in protein synthesis rates in brain. These data confirm the utility of 2-deoxyglucose measures of MW as objective biochemical indices of opiate agonist and antagonist effects in vivo.
Laboratory Investigation, Apr 15, 1993
The distribution and binding of phenytoin (PHT) were studied in rat brain using anatomically inta... more The distribution and binding of phenytoin (PHT) were studied in rat brain using anatomically intact tissue. The pattern and kinetics of PHT distribution in vivo were examined with quantitative carbon-i 4 autoradiography. Initially gray matter levels were greater than white matter levels, but after 30 mm the opposite condition was found. At a given time point, the levels of PHT among various gray matter structures as a group or among white matter structures as a group were uniform. The ratio of radio-activity in white matter to that in gray matter was 3:1 1 20 mm after injection of radiolabeled PHT. Thin-layer chromatography showed only PHT in the brain 1 5 mm after injection of PHT and both PHT and its hydroxylated metabolite [5-(p-hydroxyphenyl)-5-phenylhydantoinj 1 20 mm after injection. The proportions of PHT and 5-(p-hydroxyphenyl)-5-phenylhydantoin were the same
Quantitative Neuroanatomy in Transmitter Research, 1985
The goal of this current line of work was to develop quantitative autoradiographic methods to reg... more The goal of this current line of work was to develop quantitative autoradiographic methods to regionally quantify the binding properties (i.e. Kd and Bmax) of opiate ligands in morphine dependent and naive rat brain. Such regional studies were not possible utilizing binding techniques to brain membrane preparations because of the limited availability of tissue from dissection of small brain regions. Even with regional dissection techniques, the fine anatomical resolution available with autoradiography was sacrificed. With quantitative autoradiographic methods, one could ask whether the development of opiate tolerance/dependence occurred as a result of regulation of the affinity or concentration of opiate binding sites in any of one or several regions rich in opiate binding sites.
Blood, 1989
Human 125I-plasminogen bound readily to rat hepatocytes in primary culture at 4 degrees C and at ... more Human 125I-plasminogen bound readily to rat hepatocytes in primary culture at 4 degrees C and at 37 degrees C. Binding was inhibited by lysine and reversed by lysine, epsilon-aminocaproic acid, or nonradiolabeled plasminogen. The Kd for binding of 125I-plasminogen to hepatocytes was 0.59 +/- 0.16 mumol/L, as determined from the saturation isotherm by nonlinear regression (r2 = 0.99) and the Scatchard transformation by linear regression (r2 = 0.93). The number of sites per cell was 14.1 +/- 1.1 x 10(6). Fibrinogen synthesis and secretion by hepatocytes was insufficient to account for the major fraction of plasminogen binding, as determined by enzyme-linked immunosorbent assay (ELISA). Polyacrylamide gel electrophoresis and trichloroacetic acid precipitation studies demonstrated that plasminogen is neither activated nor degraded when bound to hepatocytes at 37 degrees C. Thin slices of whole rat liver (500 microns), isolated and prepared totally at 4 degrees C, bound 125I-plasminogen....
Transplantation Proceedings, 1998
American Journal of Clinical Pathology, 1993
Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 1993
Fifty-three cases of B-cell non-Hodgkin's lymphoma (BNHL) and 41 cases of non-BNHL lesions we... more Fifty-three cases of B-cell non-Hodgkin's lymphoma (BNHL) and 41 cases of non-BNHL lesions were retrospectively evaluated for the quantitative features of restricted surface light chain expression, pan B-cell antigens, pan T-cell antigens, and T-helper and T-suppressor phenotype using flow cytometry. Decision limit analyses were applied to multiple quantitative indices of immunophenotype to establish criteria for the detection of clonal proliferation associated with BNHL or non-BNHL conditions. Two data expressions (percent population and relative ratios) were simultaneously analyzed. The percent population measures were amenable to parametric analyses; the ratio data were amenable to nonparametric analyses only. Acceptable diagnostic specificity and sensitivity were obtained using decision limits of 75% kappa light chain and 65% lambda light chain for the detection of clonal proliferations associated with BNHL. Comparable diagnostic criteria for light chain ratios of 3:1 and 2:...
The Journal of pharmacology and experimental therapeutics, 1983
The binding of a radiolabeled opiate agonist ([3H]etorphine) and antagonist ([3H]naloxone) was st... more The binding of a radiolabeled opiate agonist ([3H]etorphine) and antagonist ([3H]naloxone) was studied using quantitative film autoradiography of rat-brain sections labeled by in vitro dipping methods. The binding activities of both [3H]naloxone and [3H] etorphine were saturable in three brain regions: noncluster striatum, nucleus accumbens and cingulate cortex. Eadie-Hofstee analysis of these regions yielded the following binding affinities and capacities: noncluster striatum binding affinity (KD) +/- S.E. = 1.59 +/- 0.23 nM, maximal binding capacity (Bmax) +/- S.E. = 28.3 +/- 1.9 fmol/mg, S.D. error of the raw data (Erad) = 6.4%; nucleus accumbens, KD +/- S.E. = 1.74 +/- 0.28 nM, Bmax +/- S.E. = 73.3 +/- 5.2 fmol/mg, S.D. (Erad) = 6.2%; cingulate cortex, KD +/- S.E. = 1.44 +/- 0.15 nM, Bmax +/- S.E. = 37.6 +/- 1.4 fmol/mg, S.D. (Erad) = 2.5%. A KD +/- S.E. = 1.72 +/- 0.29 nM, Bmax +/- S.E. = 74.1 +/- 5.3 fmol/mg, S.D. (Erad) = 5.0% was found for [3H]etorphine binding in the nonclu...
The Journal of pharmacology and experimental therapeutics, 1987
The distribution and binding of phenytoin (PHT) were studied in rat brain using anatomically inta... more The distribution and binding of phenytoin (PHT) were studied in rat brain using anatomically intact tissue. The pattern and kinetics of PHT distribution in vivo were examined with quantitative carbon-14 autoradiography. Initially gray matter levels were greater than white matter levels, but after 30 min the opposite condition was found. At a given time point, the levels of PHT among various gray matter structures as a group or among white matter structures as a group were uniform. The ratio of radioactivity in white matter to that in gray matter was 3:1 120 min after injection of radiolabeled PHT. Thin-layer chromatography showed only PHT in the brain 15 min after injection of PHT and both PHT and its hydroxylated metabolite [5-(p-hydroxyphenyl)-5-phenylhydantoin] 120 min after injection. The proportions of PHT and 5-(p-hydroxyphenyl)-5-phenylhydantoin were the same in white as in gray matter. Direct chemical measurements of brain samples obtained after coinjections of tracer amount...
Modern Pathology, 2001
The ability to predict cancer progression may help the clinical management of patients with penil... more The ability to predict cancer progression may help the clinical management of patients with penile squamous cell carcinoma. We studied 22 cases of squamous cell carcinoma of the penis diagnosed between 1989 and 1998. The depth of invasion was measured from the basement membrane of the squamous epithelium to the deepest invasive cancer cells. Cancer progression was defined as the development of lymph node metastasis or distant metastasis. The mean patient age was 63 years and the mean follow-up was 28 months. Ten patients developed cancer progression. The mean depth of invasion among patients with cancer progression was 9.8 mM, as compared to the mean depth of invasion of 4.0 mM among those patients without cancer progression (P ؍ .02). Vascular invasion was also predictive of cancer progression (P ؍ .02). Metastases developed in the majority (6 out of 7) of cases invading more than 6 mM, but developed only in a minority (4 out of 15) of cases invading 6 mM or less. We conclude that depth of invasion and vascular invasion are significant predictors of cancer progression for penile squamous cell carcinoma.
Cell and Tissue Banking, 2001
In vitro proliferation of isolated pancreaticislets has become an area of great interest given th... more In vitro proliferation of isolated pancreaticislets has become an area of great interest given the scarcity of clinicalisletdonors and the islet mass requirements for clinical islet transplantation.Smallintestinal submucosa (SIS), a naturally occurring extracellular matrix, hasbeeninvestigated to promote wound healing, tissue remodeling and cell growth. Thisstudy evaluated recovery and function of isolated canine pancreatic isletsfollowing in vitro tissue culture. Pancreatic islets
Journal of Cutaneous Pathology, 1992
A commercially available antibody to proliferating cell nuclear antigen was used to characterize ... more A commercially available antibody to proliferating cell nuclear antigen was used to characterize and compare proliferating cell populations in paraffin sections of benign, premalignant, and malignant lesions of human epidermis using routine immunohistochemical techniques. Three patterns emerged. An ordered pattern was found in prurigo nodularis and keratoacanthoma, wherein moderately and strongly positive nuclei were distributed in a continuous, basal-suprabasal layer of relatively uniform thickness. There was graded loss and ultimate extinction of PCNA staining in progressively more superficial epidermal cells. A basal dysplastic pattern was found in actinic keratosis and squamous cell carcinoma. Nuclei of essentially all dysplastic cells of both categories expressed PCNA, with a preponderance of strongly positive nuclei. These were localized to basal-suprabasal zones that were often expanded. Loss of PCNA reactivity toward the surface was often abrupt. Bowen's disease exhibited a diffuse dysplastic pattern, wherein large numbers of moderately and strongly positive nuclei, in random array, were present in essentially full thickness distribution. In many fields, however, a layer of cytologically bland basal cells, with faint or no nuclear staining, was interposed between dysplastic epithelium and dermis. This study has demonstrated that proliferating cell populations in epidermal lesions can be assessed with simple, inexpensive methods. There were consistent differences between the proliferating cell populations of the various entities studied, differences that can be reasonably correlated with other known clinical, microscopic, and biologic features of the lesions. This technique should provide an interesting new avenue for study of diverse cutaneous diseases.
Brain Research, 1985
We have examined the saturation features (K d and Bronx) of [3H]naloxone binding in rat brain reg... more We have examined the saturation features (K d and Bronx) of [3H]naloxone binding in rat brain regions in cytoarchitectonically intact tissues of naive, morphine dependent and precipitated withdrawal states using quantitative film autoradiography. Sixty-one saturation experiments in 13 regions of naive rat brains yielded monophasic Eadie-Hofstee plots with a mean (+ S,D.) K a of 1.87 + 0.87 nM and a mean Bma x (+ S.D.) of 101 + 66 fmol/mg. The 61 K d values in naive rats described a normal distribution of regional binding affinities that may reflect the biological variation of a single high affinity binding site. Similar studies in the morphine dependent and prec~p, itated withdrawal states showed no apparent changes in either the K d or Bm~ of regional [aH]naloxone binding. The possibility that in vitro preincubation of tissue sections masked potential alterations in [aH]naloxone binding was examined in competition studies of both morphine and naloxone for 2.0 nM [3H]naloxone binding to striatal sections. Preincubation had no effect on the ability of either opiate agonist or antagonist to compete for [3H]naloxone binding in the naive, morphine dependent or precipitated withdrawal states. The regional data comparing K d and Bma x for [3H]naloxone binding in naive, morphine dependent and precipitated withdrawal states eliminated the possibility that previous efforts to demonstrate alterations in opiate binding were confounded by homogenization and/or limitations of gross dissection. Our data suggest that either potential changes in antagonist binding in dependent and withdrawal states can only be demonstrated under strict in VlVO conditions or that post-binding transduction mechanisms are the sites of adaptive changes in naloxone sensitivity in the states of opiate dependence and withdrawal. * Preliminary reports of these data were given at the 13th Society for Neuroscience Meeting (1983) and the Wenner-Gren Symposium, Stockholm, Sweden (May, 1984).
Brain Research, 1985
The details of quantitative film autoradiography for tritium using tritium plastic standards were... more The details of quantitative film autoradiography for tritium using tritium plastic standards were examined with respect to 3 issues: (1) tritium tissue equivalent (TE) calibration; (2) correction of autoabsorption differences for gray and white matter; and (3) the use of carbon-14 standard sources as a method for quantifying tritium tissue images. Both 3H-tissue and 3H-plastic sources produced linear log-log relationships of 3H-concentration (nCi/mg source weight) vs source optical density (OD) over a reproducible OD range (0.080-0.800). The curves for both 3H-tissue and 3H-plastic were parallel; uncalibrated 3H-plastic standards exhibited a 4-fold higher autoabsorption over 3H-tissue sources for OD values in the linear range. Using chloroform extraction of brains from rats treated with either [14C]deoxyglucose ([IaC]DG) or [3H]deoxyglucose ([3H]DG), we found neither isotope loss nor redistribution after defatting (30% reduction of tissue dry weight). After chloroform extraction, the OD values from both gray and white matter structures containing carbon-14 were unaltered. Gray matter OD values increased by 28.7 + 5.6% (mean + S.D.) in structures containing tritium; white matter structures containing tritium exhibited a 115.9 + 29.3% increase in OD after chloroform extraction. The increase in OD after chloroform extraction was a fixed percent for any given tritium OD value from unextracted tissue when the value was within the linear range of 0.080-0.800 OD units. The magnitude of the higher white matter autoabsorption for tritium was confirmed using tritium impregnated cow brain pastes of variable gray/white mixtures. Chloroform extraction of tissue from [3H]DG treated rats was therefore a suitable procedure for direct correction of regionally heterogeneous autoabsorption of tritium. Finally, the rates of image generation for tritium and carbon-14 sources were compared. The rate of increase of OD with increasing exposure time was found to be equal for 3H-tissue and 3H-plastic images; sources of carbon-14 in plastic, however, exhibited more accelerated rates of image generation when compared to tritium sources (i.e. 3H-and 14C-images did not covary with exposure time). The effect of non-covariance on tritium TE calibrated standards was the overestimation of OD values for 14C-plastic standards with increasing times of exposure (comparison of 4 week images to 1 week images showed errors of 35-40%). Use of carbon-14 sources to quantify tritium-generated images therefore required recalibration of laC-plastic for all exposure times of interest.
Brain Research, 1985
Direct corrections of regional differential tritium autoabsorption in quantitative autoradiograph... more Direct corrections of regional differential tritium autoabsorption in quantitative autoradiography of the central nervous system are made using chloroform extraction of brain sections from rats receiving tritiated 2-deoxyglucose in vivo. Fifty-two regional quenching coefficients varied from 30% in gray structures to 100% in white structures. These data may be used to correct for regional quenching of tritium emissions in autoradiographic studies of rat brain.
Brain Research, 1983
We have studied regional cerebral metabolism by 2-deoxyglucose autoradiography in 67 brain struct... more We have studied regional cerebral metabolism by 2-deoxyglucose autoradiography in 67 brain structures of morphine-dependent rats during fixed dose naloxone precipitated withdrawal. Behavioral indices of withdrawal were studied simultaneously in the same animals used in the cerebral metabolism studies. The effects of cumulative morphine dose (470, 1145 and 2345 mg/kg) with fixed dose (0.5 mg/kg) naloxone precipitated withdrawal upon behavioral and cerebral metabolic measures of the severity of withdrawal were compared. All morphine dependent groups studied exhibited the known behavioral sequelae of naloxone precipitated withdrawal. Qualitative withdrawal signs exhibited by all dependent groups included tachypnea, ptosis, penile erection, ejaculation, diarrhea, urination, salivation, lacrimation, rhinorrhea, and irritability. Quantitative signs of withdrawal for 470, 1145 and 2345 mg/kg cumulative morphine dose groups were as follows (mean __
Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1989
a2-Antiplasmin (a2AP), a serpin proteinase inhibitor with two methionine residues in its reactive... more a2-Antiplasmin (a2AP), a serpin proteinase inhibitor with two methionine residues in its reactive center, was treated with c/s.dicldorodiammineplatinum (||) (c/s-DDP). This compound has been utilized previously to specifically modify methionine residues. After reaction, the a2AP demonstrated decreased inhibitory activity against plasmin, min|plasmin, trypsin and chymotryps|n. The reduction in activity depended on the concentration of ds-DDP; however, the amount of activity retained by the treated a2AP was equivalent with each of the four proteinases, a2AP that was incubated with 1.0 mM c/s-DDP for 3 h at 370C was 90% inactivated. These same conditions resulted |n the binding of only 1.0-1.5 tool of platinum per tool of inhibitor. In experiments with lower concentrations of ds-DDP, the amount of incorporated platinum directly correlated with the amount of inactivated azAP (1:1 stoichiometry). Reactions and functions of azAP that do not result in protelnase inhibition were not affected by c/s-DDP. Cleavage of a2AP by elastase, which occurs near tl*~ pro~eina~e inhibition site, was unaffected. In addition, the affinity of a~AP for the K1-3 region of plasminogen remained unchanged after treatment. These data strongly suggest that the reaction of azAP with ds-DDP involves principally a single site on the inhibitor and that this site is critical for proteinase inhibitory activity. The most likely candidate is the P~ methionine which is adjacent to the peptide bonds cleaved in the proteinase inhibitory reactions but not in the elastase reaction.
Annals of Neurology, 1987
The role of the neuropeptide somatostatin in limbic seizures was studied using electrical stimula... more The role of the neuropeptide somatostatin in limbic seizures was studied using electrical stimulation of the hippocampus in kindled rats. Cysteamine, an agent which selectively and reversibly depletes brain somatostatin stores, had a biphasic action. An early proconvulsant effect was seen within a few hours, consisting of prolonged electrographic seizures in the hippocampus and more severe behavioral convulsions. A later anticonvulsant effect, maximal at 1 to 2 days and dissipating within a week, was manifested by less intense behavioral convulsions without change in the duration of electrical seizure activity. Both effects were dose-dependent. No change in afterdischarge thresholds was detected at any time after the administration of cysteamine. Intraventricular administration of somatostatin to animals with behavioral seizures attenuated by cysteamine treatment restored the responses to precysteamine levels. We conclude that somatostatin facilitates the spread of seizures over limbic circuits from a region of focal seizure initiation.
Neurochemical Research, 1987
Quantitative autoradiography of [14C]deoxyglucose, [14C]iodoantipyrine, and [14C]leucine was used... more Quantitative autoradiography of [14C]deoxyglucose, [14C]iodoantipyrine, and [14C]leucine was used to estimate regional cerebral glucose metabolism, cerebral blood flow, and cerebral protein synthesis, respectively, in rats during morphine dependence and withdrawal. Glucose metabolism was elevated in 19 of 26 selected brain regions; the elevations in glucose metabolism were similar when data were expressed as either optical density ratios or as calculated rate values of mumol/100 gm/min. Restraining the rats produced heterogeneous effects on glucose metabolism during morphine withdrawal (MW). Neither estimated cerebral blood flow nor cerebral protein synthesis were affected by morphine and/or naloxone treatments in either naive or morphine-dependent rats. The data demonstrate that changes in regional cerebral glucose utilization occur independently of blood flow changes and exclude the possibility that regional changes in glucose utilization occur as a consequence of large regional changes in protein synthesis rates in brain. These data confirm the utility of 2-deoxyglucose measures of MW as objective biochemical indices of opiate agonist and antagonist effects in vivo.