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PubMed1981

Chemical test for mammalian feces: collaborative study.

A chemical method was developed to determine the presence of mammalian feces in food. The method consisted of the colorimetric determination of alkaline phosphatase and involved the splitting of the phosphate radical and phenolphthalein diphosphate and the resultant color development of phenolphthalein in a basic medium. A collaborative study testing the feces of 22 animals resulted in a 95% correct identification of samples as mammalian feces. The method has been adopted official first action.

Alkaline Phosphatase↗
PubMed1994

Protection of carbon tetrachloride hepatotoxicity by cinchona alkaloids.

Elevated serum enzymes were observed due to all four cinchona alkaloids in male Sprague-Dawley rats after four day treatment. Only a slight effect on bile flow and excretion of phenolphthalein glucuronide in bile was noted. Animals treated with cinchona alkaloid for four days and then receiving CCl4 showed a partial protection against CCl4 induced hepatotoxicity. The levels of serum enzymes were lowered as compared to animals treated with CCl4 alone. Bile flow and excretion of phenolphthalein glucuronide in bile showed a trend of restoration towards control levels. All four cinchona alkaloids probably inhibit microsomal enzymes, thereby, inhibiting the bioactivation of CCl4 and hence reducing the toxicity.

Alanine Transaminase↗
PubMed1985

Activities of several phase I and phase II xenobiotic biotransformation enzymes in cultured hepatocytes from male and female rats.

Hepatocytes were isolated from adult male and female rats and maintained in monolayer culture for up to 24 hr. The degree of preservation of representative phase I and phase II xenobiotic biotransformation enzymes was studied in these cells immediately after isolation, after attachment in culture, and after 24 hr in culture. Regarding phase I pathways, hepatocytes during 24 hr lost 50% of cytochrome P-450, but maintained high mixed function oxidase activities; 75% of aryl hydrocarbon hydroxylase and 65% of benzphetamine demethylase activities were preserved in hepatocytes from males, whereas in hepatocytes from females 70 and 50% of these activities, respectively, were maintained. Of phase II pathways, glutathione transferase activity after 24 hr, tested toward 1,2-dichloro-4-nitrobenzene as substrate, was diminished in male hepatocytes to 20% of the initial liver activity and in female cells, to 35%, whereas the activity tested toward 1-chloro-2,4-dinitrobenzene as substrate was stable. UDP-glucuronosyltransferase activities, tested toward p-nitrophenol and phenolphthalein as substrates, were slightly increased during 24 hr of culture of hepatocytes to levels higher than in liver before perfusion. The level of UDP-glucuronic acid, the endogenous substrate for the enzyme, was reduced after isolation to only 6% of the initial liver value, and then increased during culture to a level approximately 60% of normal. Thus, the changes in xenobiotic biotransformation enzymes and associated constituents in cultured hepatocytes were not uniform, although biotransformation capability remained reasonably intact.

Animals↗
PubMed1993

A rapid high-performance thin-layer chromatographic urine screen for laxative abuse.

Laxative abuse, if unrecognized, can lead to unnecessary and costly investigations. The stimulants and cathartics are the most commonly abused laxatives and have the potential for causing the most long-term damage. We have optimized a solid-phase extraction coupled with high-performance thin-layer chromatography (HPTLC) to provide rapid, simple, and sensitive detection of phenolphthalein, danthron, rhein, aloin, and bisacodyl and its metabolites in urine. Positive screens have revealed laxative abuse in 3 of 42 samples from patients presenting with diarrhea of unknown origin.

Anthraquinones↗
PubMed1997

Studies towards a disposable screen-printed amperometric biosensor for progesterone.

A screen-printed carbon electrode (SPCE) has been investigated as the base transducer for a disposable amperometric progesterone biosensor. The biorecognition element was a monoclonal sheep anti-progesterone antibody (mAb). This was immobilized onto the transducer by interaction with a layer of rabbit IgG which had been previously coated onto the SPCE; optimum conditions for these loadings were deduced experimentally. The device was employed in a competitive assay using alkaline phosphatase-labelled progester-one. Three possible substrates for the enzyme were considered, namely, phenyl phosphate, phenolphthalein phosphate and 4-aminophenol phosphate. Cyclic voltammetry and amperometry were carried out on the corresponding aromatic phenols and phenol itself was found to give the best electrochemical characteristics; consequently, phenyl phosphate was employed as the substrate. Chronoamperometry was used to measure the phenol produced by the reaction of bound enzyme-labelled progesterone and substrate. The chronoamperometric response was dependent on unlabelled progesterone over at least three orders of magnitude with a detection limit of about 1 x 10(-9) mol/dm3. This suggests that the device may have applications for the analysis of biological fluids.

Aminophenols↗
PubMed1997

Effect of fludrocortisone and spironolactone on sodium and potassium losses in secretory diarrhea.

The response of the colon to aldosterone is believed to be an important adaptive mechanism to excessive sodium losses in diarrhea. However, the degree to which mineralocorticoid activity actually influences fecal output of sodium in people with diarrhea is unknown. To gain insight into this question, 10 normal people were treated with placebo, fludrocortisone (an aldosterone agonist), and spironolactone (an aldosterone antagonist) during three experimental periods lasting nine days. On days 5-8, diarrhea was induced by ingestion of phenolphthalein. Diet was controlled. Fecal sodium was 40 meq/day on placebo and 29 meq/day on fludrocortisone, consistent with mineralocorticoid stimulation of intestinal sodium absorption. However, contrary to our expectations, spironolactone therapy was also associated with a fall in fecal sodium output, to 28 meq/day. To explain this paradoxical effect of spironolactone, we suggest that sodium depletion caused by spironolactone's natriuretic action on the kidney caused the release of an unknown stimulant of intestinal sodium absorption, whose action more than overcame the reduced colonic absorption resulting from inhibition of aldosterone activity by spironolactone. This interpretation implies that the intestinal adaptation to sodium depletion in diarrhea involves both aldosterone and an aldosterone independent factor, working in concert to reduce fecal sodium output.

Adult↗
PubMed1984

Elevation of beta-glucuronidase activity in medicated patients with epilepsy.

Elevations of serum beta-glucuronidase (GRS) enzyme activity can occur under a variety of pathological conditions. Using phenolphthalein glucuronic acid as the substrate, 158 epileptic patients were randomly screened for GRS. GRS was distinctly elevated (65.9 +/- 30.0 micrograms phenolphthalein/ml serum) in patients, compared with the normal group (27.0 +/- 10.0). No difference in GRS levels were found when seizure-free (greater than 1 year) patients (n = 61; GRS, 62.6 +/- 32.7 micrograms) were compared with patients who had seizure episodes within 1 week (n = 26; GRS, 73.2 +/- 24.9 micrograms), and there were no differences when intermediate groups were examined. GRS elevations were found to be linearly and directly correlated with free phenytoin ultrafiltrate levels (n = 35, r = 0.7692, p less than 0.0001) when patients were co-medicated with valproic acid, with serum phenobarbital levels (n = 58, r = 0.5361, p less than 0.05), with serum valproic acid levels (n = 43, r = 0.3173, p less than 0.05), and with the sum of serum phenobarbital and valproic acid levels (n = 16, r = 0.8657, p less than 0.0001). The findings indicate that GRS elevations are probably due to anticonvulsant medications rather than to the frequency of seizures. There is no evidence that GRS determinations can be used for the diagnosis or prognosis of patients with epilepsy.

Adolescent↗
PubMed1991

Glucuronidation and sulfation in subcellular fractions and in the isolated perfused rabbit lung: influence of ethanol.

Glucuronidation and sulfation of 1-naphthol, 7-hydroxycoumarin, 4-nitrocatechol and phenolphthalein were studied in rabbit lung and liver. Pulmonary UDP-glucuronyltransferase and sulfotransferase activities in subcellular fractions were approximately 20-50% of those determined in the liver. Ethanol did not markedly induce these enzymes in either tissue. Glucuronidation and sulfation of 1-naphthol and 7-hydroxycoumarin were also studied in the isolated perfused rabbit lung as an intact cell model. Neither glucuronidation nor sulfation of 1-naphthol was observed. The absence of conjugate formation was due neither to the presence of beta-glucuronidase and/or sulfatase, nor to alternative biotransformation pathways. About 35% of the initial 7-hydroxycoumarin was conjugated, the majority being sulfate conjugate (14.4 nmol/h) with only minor amounts (0.12%) of the glucuronide. These results indicate the importance of studying both whole organ and in vitro metabolism.

Animals↗
PubMed1983

Purging and calorie absorption in bulimic patients and normal women.

Self-induced purging with laxatives is common among bulimic persons, who assume that purging reduces intestinal absorption of ingested calories. However, the efficacy of purging in reducing calorie absorption has never been studied, probably because the standard calorie balance procedure is expensive and time consuming. With a recently devised method, calorie absorption during a single day was measured to determine to what extent phenolphthalein or saline purge reduced calorie absorption. In two bulimic patients who regularly used laxatives for weight control and five normal young women, even extreme purging producing 4 to 6 L of diarrhea caused calorie absorption to decrease by only about 12% of calorie intake. The theoretical basis on which laxatives are taken for weight control is unsound.

Adult↗
PubMedDate not supplied

Development of UDP-glucuronosyltransferase activity toward digitoxigenin-monodigitoxoside in neonatal rats.

Glucuronidation is low or undetectable in embryonic and early fetal tissues and changes to adult levels at rates depending on the acceptor, tissue, and species. Because other data indicate there may be a specific UDP-glucuronosyltransferase (GT) in the liver of adult rats that glucuronidates digitoxigenin-monodigitoxoside (DIG), the development of GT activity in neonatal rats toward DIG was compared with that of other acceptors. Conjugation of p-nitrophenol and 1-naphthol was higher at birth and decreased to adult levels by 20 days of age. Glucuronidation of chloramphenicol, morphine, valproic acid, and bilirubin increased from birth to adult activity by 20 days of age. Conjugation of phenolphthalein, estrone, and diethylstilbestrol was low in 1-day-old rats and higher than adult in 20-day-old animals. In contrast, glucuronidation of DIG was barely detectable (9% of adult) in 20-day-old rats. The concentration of UDP-glucuronic acid was 50% of adult levels at birth and increased to adult values by 10 days of age. Administration of 3-methylcholanthrene on days 6 to 9 after birth significantly stimulated GT activity toward 1-naphthol, p-nitrophenol, and morphine, whereas phenobarbital precociously increased conjugation of chloramphenicol, valproic acid, morphine, and diethylstilbestrol. Pregnenolone-16 alpha-carbonitrile enhanced the development of GT activity toward morphine, chloramphenicol, valproic acid, bilirubin, diethylstilbestrol, and estrone. Glucuronidation of DIG was not increased after 3-methylcholanthrene or phenobarbital, but could be induced after pregnenolone-16 alpha-carbonitrile to 7% of adult values in 10-day-old rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMedDate not supplied

A comparison of the presumptive luminol test for blood with four non-chemiluminescent forensic techniques.

Presumptive blood detection tests are used by forensic investigators to detect trace amounts of blood or to investigate suspicious stains. Through the years, a number of articles have been published on the popular techniques of the day. However, there is no single paper that critiques and compares the five most common presumptive blood detection tests currently in use: luminol, phenolphthalein (Kastle-Meyer), leucomalachite green, Hemastix and the forensic light source. The present authors aimed to compare the above techniques with regard to their sensitivity, ease of use and safety. The luminol test was determined to be the most sensitive of the techniques, while Hemastix is a suitable alternative when the luminol test is not appropriate.

Benzidines↗
PubMed1975

Choleresis and hepatic transport mechanisms. II. Influence of bile salt choleresis and biliary micelle binding on biliary excretion of various organic anions.

To investigate, whether binding to micelles has a function in hepatic transport, biliary excretion of three organic anions, phenolphthalein-beta-D-glucuronide (PG), dibromosulphthalein (DBSP) and indocyanine green (ICG) was studied in rats during saline, taurocholate or dehydrocholate administration. Taurocholate causes a weak choleresis with formation of biliary micelles, dehydrocholate a strong choleresis with little micelle formation. The two bile salts did not uniformly influence biliary excretion of the organic anions: biliary excretion of ICG (12.9 mumoles/kg) and DBSP (75.0 mumoles/kg) was stimulated by both bile salts: ICG excretion most pronounced by taurocholate and DBSP excretion most strongly by dehydrocholate. Biliary output of PG (25.8 and 200 mumoles/kg) was not stimulated by bile salt administration. Binding of PG, DBSP and ICG to biliary micelles was studied in sedimentation experiments by ultracentrifugation. PG, DBSP and ICG in bile showed a similar sedimentation pattern as 3H-taurocholate in bile, which indicates an association of all three anions with biliary micelles. Thus, the influence of bile salts on biliary transport of organic anions varies with the compound studied and the bile salt used, effects which cannot be explained by differences in binding to biliary micelles.

Animals↗
PubMed1978

Is the effect of diphenolic laxatives mediated via release of prostaglandin E?

The diphenolic laxatives, bisacodyl and phenolphthalein, and the osmotic laxative mannitol increase intestinal fluid volume in the rat colon in situ. The diphenolic laxatives stimulate the biosynthesis of prostaglandin E (PGE) whereas mannitol does not. Inhibition of PG-biosynthesis by pretreatment with indomethacin only reduces the effect of the diphenolic laxatives. It is suggested that diphenolic laxatives increase intestinal fluid volume via stimulation of PGE biosynthesis in the colon.

Animals↗
PubMed1979

The origins of unconjugated bilirubin in bile.

In order to investigate the origin of unconjugated bilirubin in bile, beta-glucuronidase activity in rat and human bile was determined at various pH. beta-Glucuronidase in rat and human bile had their optimum pH at 5.5 when phenolphthalein glucuronide and delta 1-azopigment were used as substrate, and at 6.0 when bile itself was incubated. In human and rat bile the hydrolysis was suppressed to a minimum at each physiologic pH. However, human bile shows remarkable hydrolysis in alkaline pH (7.5--8.0). On the other hand, when delta 1-azopigment was incubated in various buffers, several per cent of delta 1-azopigment were hydrolyzed non-enzymatically in neutral to alkaline pH. Thus, it was suggested that enzymatic and nonenzymatic hydrolysis contributes to the existence of unconjugated bilirubin in bile.

Animals↗
PubMed2006

Identification of the human liver UDP-glucuronosyltransferase involved in the metabolism of p-ethoxyphenylurea (dulcin).

Dulcin (DL), now banned, was once a widely used artificial sweetener. DL possesses an ureido group that is metabolized by direct glucuronidation in rabbit liver microsomes. Dulcin N-glucuronide (DNG) is the only type of ureido N-glucuronide known to date; ureido glucuronidation in humans has not been previously reported. Accordingly, the glucuronidation of DL was studied using human liver microsomes (HLM) and expressed human UDP-glucuronosyltransferase (UGT) enzymes. The average K (m) and V (max) values from nine HLM samples were 2.10 mM and 0.156 nmol/mg/min, respectively. Of the six human UGT isoforms screened for their ability to glucuronidate DL, only UGT1A1 and UGT1A9 showed activity. The apparent K (m) values using UGT1A1 and UGT1A9 were 5.06 and 6.99 mM, and the apparent V (max) values were 0.0461 and 0.106 nmol/min/mg, respectively. Phenolphthalein, a substrate for UGT1A9, inhibited DL glucuronidation in HLM competitively (K (i) = 0.356 mM), but bilirubin, a substrate for UGT1A1, did not. These results suggest that UGT1A9 is a key enzyme catalyzing the glucuronidation of DL.

Bilirubin↗
PubMed2004

Biliary excretion of bile acids and organic anions in rats with dichloroethylene-induced bile canalicular injury.

BACKGROUND: Hepatocytes in zone 1 of the hepatic lobule play a role in the uptake and biliary excretion of bile acids and organic anions under physiological conditions, and hepatocytes in zone 3 may play a role only when there is a high-dose load. To further elucidate the role of hepatic zonation in the hepatic handling of bile acids and organic anions, the biliary excretion of these compounds was studied in rats with dichloroethylene (DCE)-induced selective zone 3 bile canalicular injury. METHODS: Biliary excretion of various bile acids and organic anions was studied in rats 1 h after oral administration of DCE (5 mg/100 g). The effect of DCE on the immunostaining of multidrug resistance protein 2 (Mrp2; an important canalicular organic anion transporter) in the liver was also examined. RESULTS: The biliary excretory maximum of taurocholate and tauroursodeoxycholate was decreased in DCE-treated rats, whereas the biliary excretion of taurolithocholate-sulfate and phenolphthalein-glucuronide was unchanged in DCE-treated rats, and DCE treatment decreased the biliary excretion of sulfobromophthalein and pravastatin. DCE decreased Mrp2 staining in the canalicular membrane of zone 3 hepatocytes on immunohistochemistry. CONCLUSIONS: These findings indicate that canalicular transport in zone 3 hepatocytes is important in the biliary excretion of bile acids and organic anions, when they are administered at high doses.

ATP-Binding Cassette Transporters↗
PubMed1985

Assay of beta-glucuronidase in bile following ion-pair extraction of pigments and bile acids.

A method for improving the assay of beta-glucuronidase in hepatic and gallbladder bile is described. The method uses ion-pair extraction with N,N,N-triheptyl-1-heptanaminium bromide to remove pigments and bile acids. Conjugated bilirubin, unconjugated bilirubin, and taurine and glycine conjugates of deoxycholic and chenodeoxycholic acids are extracted efficiently from bile by the procedure. The sensitivity of the spectrophotometric assay of beta-glucuronidase in bile using phenolphthalein glucuronide is increased significantly.

Animals↗
PubMed1975

Effect of fasting on substrate specificity of rat liver UDP-glucuronosyltransferase.

The effect of a 3-day period of complete starvation on the hepatic UDPglucuronosyltransferase activity was studied in the rat. The substrate specificity of the enzyme was assayed with bilirubin as a carboxylic acceptor, and phenolphthalein and p-nitrophenol as phenolic acceptors. Starvation increased the bilirubin UDPglucuronosyltransferase specific activity by 33%, whereas no increase in specific activities appeared when the phenolic substrates were used. However, on a total liver weight basis, all three activities were significantly lower than those of the controls. Kinetic studies of activated microsomal bilirubin UDPglucuronosyltransferase showed that apparent Km values were similar; fasting acted only by increasing V. The results suggest that the changes in bilirubin glucoronosyltransferase activity provoked by starvation may reflect actual enzyme induction; they favour the multiplicity of the UDPglucuronosyltransferase system.

Animals↗