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Biomedical subjects

E R Gordon

Publications and source records attributed to E R Gordon.

At least 19 recordsLinked to original sources

Animal-based national surveillance for zoonotic disease: quality, limitations, and implications of a model system for monitoring rabies.

Surveillance for zoonotic diseases among wildlife is a research and public health challenge. The inherent limitations posed by the requisite human-animal interactions are often undefined and underappreciated. The national surveillance system for animal rabies in the United States was examined as a model system; reporting of animal rabies is legally mandated, each case of rabies is laboratory confirmed, and data have been consistently collected for more than 50 years. Factors influencing the monthly counts of animal rabies tests reported during 1992-2001 were assessed by univariate and multivariable regression methods. The suitability of passively collected surveillance data for determining the presence or absence of the raccoon-associated variant of rabies within states and within individual counties was assessed by determining critical threshold values from the regression analyses. The size of the human population and total expenditures within a county accounted for 72% and 67%, respectively, of the variance in testing. The annual median number of rabies tests performed was seven for counties without rabies, 22 for counties with non-raccoon rabies, and 34 for counties with raccoon rabies. Active surveillance may be required in locales with sparse human populations when a high degree of confidence in the status of rabies is required.

Animals↗

Persistence of elevated rabies prevention costs following post-epizootic declines in rates of rabies among raccoons (Procyon lotor).

Determining the benefits to cost relationships among different approaches to rabies control and prevention has been hindered by the inherent temporal variability in the dynamics of disease among wildlife reservoir hosts and a tangible and objective measure of the cost of rabies prevention. A major and unavoidable component of rabies prevention programs involves diagnostic testing of animals and the subsequent initiation of appropriate public health responses. The unit cost per negative and positive diagnostic test outcome can be reasonably estimated. This metric when linked to methodologies subdividing the epizootic process into distinct temporal stages provided the requisite detail to estimate benefits derived from rabies control strategies. Oral rabies vaccine (ORV), for prevention of the raccoon-associated variant of rabies, has been distributed in Ohio and adjoining states in an effort to develop an immune barrier to the westward spread of epizootic raccoon rabies. The costs of ORV delivery have been quantified. Herein, the cost structures required to assess the benefits accrued by prevention were developed. A regression model was developed effectively predicting (r2=0.70) the total number of rabies diagnostic tests performed by 53 counties in five northeastern (NE) states from 1992 to 2001. Five temporal stages sufficed to capture the range of variability in the raccoon rabies epizootic process. Unit costs, dollars per diagnostic test outcome, were calculated for negative and positive results from published reports. Ohio counties were matched to NE counties based on similar socioeconomic characters. A "pseudo-epizootic" of raccoon rabies was introduced into Ohio and the costs savings from ORV were derived as the excess costs imposed by epizootic spread throughout the state. At 46 km/year (range modeled, 30-60 km/year), the pseudo epizootic spread, and reached the enzootic stage, in all Ohio counties by year 13 (range modeled, 11-17 years). Cumulative excess costs for Ohio ranged between $11 and $21 million; counties of low socioeconomic status experienced the greatest relative excess costs. The costs for rabies prevention activities reached apices during the epizootic stage of raccoon rabies (2.7-10.8 times baseline) an unforeseen finding indicated elevated costs persisted (1.7-7.2 times baseline) into the enzootic stage.

Animals↗

Temporal dynamics of rabies in a wildlife host and the risk of cross-species transmission.

An epidemiological model was developed for rabies, linking the risk of disease in a secondary species (cats) to the temporal dynamics of disease in a wildlife reservoir (raccoons). Data were obtained from cats, raccoons, and skunks tested for rabies in the northeastern United States during 1992-2000. An epizootic algorithm defined a time-series of successive intervals of epizootic and inter-epizootic raccoon rabies. The odds of diagnosing a rabid cat during the first epizootic of raccoon rabies was 12 times greater than for the period prior to epizootic emergence. After the first raccoon epizootic, the risk for cat rabies remained elevated at levels six- to seven-fold above baseline. Increased monthly counts of rabid raccoons and skunks and decreasing human population density increased the probability of cat rabies in most models. Forecasting of the public health and veterinary burden of rabies and assessing the economics of control programmes, requires linking outcomes to dynamic, but predictable, changes in the temporal evolution of rabies epizootics.

Animals↗

VDP survey.

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Education, Dental↗

Accumulation of unconjugated bilirubin in cholesterol pellets implanted in swine gallbladders.

BACKGROUND & AIMS: Most cholesterol gallstones have a pigmented center, but it is unclear whether its presence is primary or secondary. This study was performed to determine if bilirubin would accumulate in a gallstone model consisting of cholesterol pellets. METHODS: Cholesterol was compressed into pellets at 2500 psi, producing a pellet that behaved like human cholesterol gallstones in regard to penetration of solutes into the stone. Pellets were implanted into gallbladders of pigs and harvested after 4 weeks. Bilirubin species were measured by high-performance liquid chromatography. RESULTS: The proportions of bilirubin species in bile were not changed by the presence of pellets, i.e., diconjugates (mean +/- SD, 1.9% +/- 1.0% vs. 0.7% +/- 0.8%), monoconjugates (83.8% +/- 5.5% vs. 87.8% +/- 6.6%), and unconjugated bilirubin (14.2% +/- 5.3% vs. 11.5% +/- 5.6%) were similar at the time of implantation and removal. The cut surfaces of the pellets were pigmented. Pellets contained 5.46 +/- 1.38 micrograms bilirubin/g sample at harvesting, and 98.6% +/- 2.3% of bilirubin in pellets was unconjugated. In in vitro studies, there was a large increase in unconjugated bilirubin in the bile. Pellets also became pigmented in vitro, but there was considerable variability in the bilirubin species present in the pellets. CONCLUSIONS: Unconjugated bilirubin accumulates in cholesterol pellets and pigments them. This provides a mechanism by which cholesterol gallstones could become secondarily pigmented.

Analysis of Variance↗

Serum bilirubin pigments covalently linked to albumin.

The serum levels of biliprotein (i.e., the bilirubin fraction covalently linked to albumin) were measured in patients with hepatobiliary disease by two analytical procedures: reverse-phase anion-exchange chromatography (Bond-Elut) and anion-exchange chromatography. Bond-Elut extracts contained tetrapyrroles, allowing the analysis of biliprotein, bilirubin, and bilirubin conjugates, whereas the anion-exchange method converted tetrapyrroles to azodipyrroles (protein-free), which were adsorbed on the resin, with the exception of azopyrrole of biliprotein (covalently linked to albumin), which passed through the resin and could be analyzed. In contrast, other cationic polymer columns did not completely bind the alkaline diazo derivatives of unconjugated bilirubin, conjugated bilirubin, or the azopyrrole of biliprotein not bound to albumin. Therefore, a comparison was made of these two methods using the original anion-exchange resin. Both procedures gave similar values for serum levels of biliprotein in patients with high levels of total bilirubin.

Bile Pigments↗

Bilirubin conjugate changes in the bile of gallbladders containing gallstones.

Gallbladder bile was obtained at laparoscopic cholecystectomy from 31 patients with gallstones, and duodenal aspirates from 18 normal controls. Bile pigments (9 conjugates and unconjugated bilirubin) were analyzed by high-performance liquid chromatography. The average proportional composition of the bile pigments from the patients with gallstones was characteristically different from the controls. Whereas the average values for the principal conjugates in the controls were bilirubin diglucuronide 83.4%, bilirubin monoglucuronide 10.1%, bilirubin monoglucuronide monoglucoside 4.5%, and bilirubin monoglucuronide monoxyloside 1.0%, the corresponding values in the biles from the patients with gallstones were 66.3%, 20.6%, 6.5%, and 2.8%, respectively. Values from the more minor conjugates and unconjugated bilirubin were less than 2% in either data set. In samples obtained in 9 of the gallstone patients early and late in the procedure, no significant change was found. Over the spectrum of findings in the gallstone patients, as the proportion of bilirubin diglucuronide became smaller, that of bilirubin monoglucuronide increased substantially, whereas those of bilirubin monoglucuronide monoglucoside and bilirubin monoglucuronide monoxyloside increased to a small extent. The findings suggest that bilirubin diglucuronide hydrolysis occurs in the gallbladder bile of gallstone patients, with the production of bilirubin monoglucuronide, and that if further hydrolysis of bilirubin monoglucuronide occurs with the formation of unconjugated bilirubin, the latter does not ordinarily increase because it is being absorbed. Stasis with increased gallbladder residence time was likely present in some of the patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Bile↗

Changes in bilirubin pigments secreted in bile after liver transplantation.

The species of bile pigments secreted in T-tube fistula bile after liver transplantation were ascertained by high-performance liquid chromatography in 15 patients for 10 days after liver transplant. Nine glycosidic conjugates and unconjugated bilirubin were resolved by the analytical procedure. The principal pigments in bile and their proportions in normal patients were the following: bilirubin diglucuronide = 83.0% +/- 3.1% (S.D.); bilirubin monoglucuronide = 9.7% +/- 1.4% (S.D.); bilirubin monoglucuronide monoglucoside = 4.0% +/- 2.8% (S.D.); and bilirubin monoglucuronide monoxyloside = 1.5% +/- 1.8% (S.D.). All of the other possible glucuronide, glucose and xylose monoconjugates and diconjugates and unconjugated bilirubin were also found, but each was normally less than 1% of the total. In 13 of the 15 transplant patients, a significant depression in proportions of bilirubin diglucuronide and elevation in proportions of bilirubin monoglucuronide were found after the transplant, with an accompanying but generally small increase in the proportions of the minor conjugates. In two patients with rejection of the transplant, the changes were of larger magnitude, with improvement occurring only with recovery from the rejection. In one of these patients, kidney failure was present, and in addition to the diglucuronide and monoglucuronide conjugates, diglucoside and monoglucoside monoxyloside conjugates were found in plasma. The underlying metabolic abnormalities are not clear but likely reflect underlying abnormal intracellular cofactor levels for conjugation. Glycogen depletion with reduction of UDP-glucuronate levels or reduced UDP-glucuronate formation from UDP-glucose, secondary to elevation of UDP-xylose, could potentially account for the changes in pigment excretion.

Adult↗

High-performance liquid chromatographic separation of bilirubin conjugates: the effects of change in molarity and pH.

A fast, sensitive high-performance liquid chromatographic method has been developed for the separation and quantitation of biliary bile pigments; this utilizes a C18 reversed-phase column with two solvents, a buffer and an organic solvent, which were changed in a linear gradient from a polar to a less polar combination. Nine glycosidic conjugates of bilirubin as well as unconjugated bilirubin and a suitable internal standard, unconjugated mesobilirubin IX alpha, were all separated to baseline by gradient elution; the species eluted in a polar to less polar fashion. Increasing the molarity of the solvent decreased the binding of non-glucuronide pigments to the column, with a decrease in their retention times, whereas for bilirubin monoglucuronide they increased. Decrease in pH, similarly, preferentially increased bilirubin monoglucuronide retention times.

Animals↗

Cryptic Na+,K(+)-ATPase activity in rat liver canalicular plasma membranes: evidence for its basolateral origin.

Controversy exists concerning the localization of the enzyme Na+,K(+)-ATPase to canalicular membranes in hepatocytes. Most studies find enzyme activity only at the basolateral plasma membrane domain of the hepatocyte. However, Na+,K(+)-ATPase activity has been detected recently in a canalicular membrane fraction prepared by Mg++ precipitation, suggesting that differences in membrane domain fluidity account for these discrepancies. To reinvestigate this question, we used free-flow electrophoresis to further purify canalicular liver plasma membranes originally separated by sucrose density centrifugation. With this technique, canalicular membranes devoid of Na+,K(+)-ATPase activity by routine assay were separated into six subfractions. More than 80% of the activities of canalicular marker enzymes was recovered in two subfractions closest to the anode, which were totally devoid of Na+,K(+)-ATPase activity. However, Na+,K(+)-ATPase activity could now be detected in the four other fractions that contained only small amounts of canalicular marker enzymes. The basolateral marker enzyme, glucagon-stimulated adenyl cyclase, comigrated with this cryptic Na+,K(+)-ATPase activity. Furthermore, addition of 6 mumol/L [12-(2-methoxyethoxy)-ethyl-8-(cis-2-n-octylcyclopropyl)-octanoate ], a membrane-fluidizing agent, to the original canalicular membrane preparation and to all subfractions did not stimulate or unmask latent Na+,K(+)-ATPase activity. Finally, when canalicular membranes isolated by Mg++ precipitation were subjected to free-flow electrophoresis, they could not be separated from the more positively charged Na+,K(+)-ATPase-containing fractions, probably because of alterations in surface charge. Together these findings suggest that Na+,K(+)-ATPase is a basolateral enzyme, that represents a small contaminant when present in canalicular liver plasma membranes and that methodological differences may account for previous discrepancies.

Animals↗

Protein kinase C agonists inhibit bile secretion independently of effects on the microcirculation in the isolated perfused rat liver.

The role of hormones in the regulation of bile secretion is not known; however vasoactive agents, which act via the phosphoinositide signal transduction pathway, may mediate changes in bile flow by altering the hepatic microvasculature. We therefore examined the effects of phorbol esters and diacylglycerol, agonists of the protein kinase C branch of the phosphoinositide cascade, on perfusion pressure and bile flow in a single-pass, hemoglobin-free, isolated perfused rat liver system with constant perfusate flow. The active phorbol ester, 12,13-phorbol dibutyrate, produced a dose-dependent (maximal effect at 10(-6) M), sustained and reversible decrease in bile flow from 1.09 +/- 0.18 to 0.61 +/- 0.09 microliters per min per gm liver (37.2 +/- 5.9%) while simultaneously increasing perfusion pressure from 12.3 +/- 0.7 to 21.5 +/- 2.5 cm H2O (74.0 +/- 4.3%). Both effects were inhibited by the synthetic protein kinase C antagonist H-7. 1,2-Dioctanoyl-sn-glycerol, a diacylglycerol, produced changes in bile flow and perfusion pressure that were similar to, but more marked than, those caused by 12,13-phorbol dibutyrate, whereas the inactive phorbol ester 4 alpha-phorbol didecanoate and the vehicle dimethyl sulfoxide had no effects on either parameter. 12,13-Phorbol dibutyrate infusion resulted in reversible decreases in oxygen consumption (23.3%) and a reversible vascular redistribution of trypan blue dye but did not alter hepatic venous effluent concentrations of K+.(ABSTRACT TRUNCATED AT 250 WORDS)

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

High-performance liquid chromatographic separation of bilirubin conjugates.

A fast sensitive method for the isolation and quantitation of biliary bile pigments by reverse-phase high-performance liquid chromatography has been developed. Nine conjugates of bilirubin as well as unconjugated bilirubin and an internal standard, unconjugated mesobilirubin IX alpha, were all separated to baseline by gradient elution. The following sequence of eluted compounds was chemically identified by separating their ethyl anthranilate derivatives by thin-layer chromatography and by their enzymatic formation with UDP-bilirubin transferase and cosubstrate: bilirubin diglucuronide, bilirubin monoglucuronide monoglucoside, bilirubin monoglucuronide monoxyloside, bilirubin monoglucuronide (C-8, C-12), bilirubin diglucoside, bilirubin monoglucoside monoxyloside, bilirubin dixyloside, bilirubin monoglucoside (C-8, C-12), and bilirubin monoxyloside. The use of the commercially available mesobilirubin IX alpha as an internal standard was found to facilitate quantitation of the bilirubin conjugates.

Animals↗

Role of liver plasma membrane fluidity in the pathogenesis of estrogen-induced cholestasis.

The role of liver plasma membrane (LPM) fluidity in the pathogenesis of intrahepatic cholestasis in rats was assessed by comparing the effects of ethinyl estradiol, a cholestatic agent, and spironolactone on membrane fluidity and bile flow. Spironolactone is a steroid that has some feminizing actions but that lacks the phenolic A ring necessary for estrogens to cause cholestasis. Bile flow was reduced 42% (p less than 0.01) by ethinyl estradiol and increased 22% (p less than 0.05) by spironolactone; however, both agents produced a significant reduction of membrane Na+, K+-ATPase activity (p less than 0.01) and fluidity (p less than 0.01). The decreased fluidity persisted in liposomes prepared from the total lipid extract as well as the phospholipid extract of these membranes. Both agents produced similar significant increases in the cholesterol ester content and cholesterol-to-phospholipid molar ratio of the membranes. In addition, ethinyl estradiol and spironolactone increased the membrane sphingomyelin content (15% and 11%, respectively); however, neither agent altered the fatty acid composition of the phospholipids. Because the decreased fluidity persisted in liposomes prepared from phospholipids extracted from the LPMs of treated rats, changes in membrane cholesterol are not the sole cause of the altered membrane fluidity. Rather, the increased sphingomyelin is at least partially responsible for these changes. Also, because ethinyl estradiol and spironolactone produce similar changes in LPM lipid composition and fluidity but disparate effects on bile flow, membrane fluidity as assessed by fluorescence polarization does not appear to be the rate-limiting determinant of bile flow in estrogen-induced cholestasis.

Animals↗

Isolation and characterization of a polarized isolated hepatocyte preparation in the skate Raja erinacea.

Hepatocytes of the small skate (Raja erinacea) were isolated by collagenase perfusion and evaluated by a variety of functional and morphologic criteria. Cell yield was 1.45 X 10(8) +/- 1.3 X 10(7) cells per isolation, and as long as 8 h after isolation 98% of the hepatocytes excluded Trypan blue and no leakage of lactate dehydrogenase (LDH) or cell associated potassium could be detected. Oxygen consumption averaged 1.6 +/- 0.5 nmol/min/mg cell protein, was not stimulated by 1 mM succinate, and also remained stable for up to 8 h following isolation. However, 2,4,-dinitrophenol (5 X 10(-5) M) produced a 55% increase in oxygen utilization while ouabain, (1 mM) or sodium removal decreased oxygen consumption by 31 +/- 6 and 33 +/- 7%, respectively, indicating that a significant portion of the cells energy utilization is coupled to the activity of plasma membrane Na+, K+-ATPase. Light microscopic studies showed that the individual hepatocytes had diameters of 28 +/- 5 microns and contained large lipid droplets. Electron microscopy revealed groups of three to five cells with normal ultrastructure and tight junctions and desmosomes surrounding a single bile canaliculus. These studies indicate that skate hepatocytes can be isolated in high yield that retain their structural polarity in the form of clusters of cells formed around a single bile canaliculus. These hepatocytes remain morphologically intact and metabolically stable for a prolonged period of time.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The excretion pattern of biliverdin and bilirubin in bile of the small skate (Raja erinacea).

Bile pigment composition (biliverdin, bilirubin and their conjugates) was analyzed in stored gallbladder bile and newly synthesized hepatic bile from the small skate (Raja erinacea). During a five day period of captivity, gallbladder volume remained relatively constant while bilirubin and biliverdin content increased two to three fold. Biliverdin which accounted for 50% of the pigments did not increase as a percentage of tetrapyrroles during this period. The relative proportion of bilirubin and its conjugates also remained constant, averaging 65% for bilirubin monoglucuronide, 30% for bilirubin diglucuronide and 5% for unconjugated bilirubin as measured by HPLC methods. Intravenous administration of biliverdin resulted in significant increases in the biliary excretion of both biliverdin and all bilirubin tetrapyrroles. Insignificant quantities of 3H-biliverdin were detected in hepatic bile following the intravenous administration of 3H-bilirubin. These studies indicate that the small skate excreted both biliverdin and bilirubin conjugates in bile and that the biliverdin was not produced by in vitro oxidation of bilirubin or its metabolites.

Animals↗

Role of the physical state of the hepatic microsomal membrane in the formation of bilirubin diglucuronide.

In vitro formation of bilirubin diglucuronide by rat hepatic microsomes proceeds efficiently only under specific conditions; i.e., a low level of bilirubin, temperature greater than 23-24 degrees C and treatment of the microsomes with a very specific level of perturbant (J. Biol. Chem., 258: 15028-15036). In the present study, the effect of temperature and detergents on the anisotropy of fluorescent probes in the rat hepatic microsomal membrane was used to determine the role of the physical state of the membrane in controlling the formation of bilirubin diglucuronide. A lipid phase separation that occurred at 23 +/- 2 degrees C was identified in these membranes indicating that bilirubin diglucuronide is formed efficiently only above the lipid phase separation when the phospholipids are in the liquid crystalline state. In addition, use of fluorescent probes for the surface and core of the membrane indicates that alterations in the physical state of the hydrophobic core rather than the phospholipid polar head group region of the membrane controls the in vitro formation of bilirubin diglucuronide.

Animals↗