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Biomedical subjects

T D Phillips

Publications and source records attributed to T D Phillips.

At least 109 records · Page 6Linked to original sources

Toxicity of ochratoxin A and tannic acid to growing chicks.

The effects of ochratoxin A (OA) and tannic acid (TA) on growing chicks were determined. One-day-old male broiler chicks were fed a diet containing the following additives for 26 days: A) none; B) 3.0 ppm OA; C) 1.5% TA; D) 3.0 ppm OA plus 1.5% TA. When compared to the controls, body weights and feed efficiencies were significantly depressed in the OA and TA groups. There was a further depression in body weights and a dramatic depression of feed efficiency in the OA-TA combination group. Pigmentation, as measured by visual shank scores, was reduced in chicks fed OA singly or in combination with TA but was not affected by feeding TA singly. There were no consistent treatment differences in the relative weights of the kidney, gizzard, proventriculus, liver, bursa, or pancreas, although there was a trend toward an elevated relative kidney weight in the groups receiving OA. Serum uric acid levels were significantly elevated in the OA and the OA-TA combination group indicating impaired renal excretory function attributable to OA. Total serum protein levels were significantly depressed in the groups receiving OA, and serum calcium levels were depressed in all treatment groups. Serum phosphorus levels were decreased in the OA and OA-TA groups but were only decreased significantly in the OA group. There were no consistent treatment differences in the hematology and other blood chemistry and mineral values.

Animals↗

High pressure liquid chromatographic determination of zearalenone in chicken tissues.

A method is reported for the extraction and analysis of zearalenone in chicken fat, heart muscle, and kidney tissue by using high pressure liquid chromatography (HPLC). Zearalenone is extracted with acetonitrile, cleaned up with hexane, and extracted further with ethyl acetate. Zearalenone is determined by HPLC using a reverse phase radial compression separation system, an ultraviolet absorbance detector, and a mobile phase of acetonitrile-water (60 + 40) (v/v). Recoveries of zearalenone added at levels from 50 to 200 ng/g are in the range 82.6-95.1%.

Adipose Tissue↗

Vanadium: chemistry and the kidney.

Vanadium (V), a metallic element of the first transition series, is widely distributed in the environment. Although an essential trace element in higher animals, chronic exposure to V is of concern because of its increased concentration near industrial operations, its occurrence in the ash of combustion products of petroleum and coal, and its subsequent biomagnification in the environment. V is found in trace amounts in both terrestrial and aquatic animals and in solution can form inorganic orthovanadate oxyanions that, if absorbed, are eliminated primarily by the kidneys. Because V is often found concentrated in renal tissue to the largest extent in the body, the kidneys may represent a major site of action. Moreover, V in the vanadate configuration increases the urinary excretion of solutes and water in the rat, and inhibits renal organic ion accumulation and renal Na+, K+-ATPase in vitro and in vivo. Furthermore, as a nutritionally required element, V may play a regulatory role in salt and water excretion by modification of the Na+ pump in the kidney.

Animals↗

High pressure liquid chromatographic determination of an O-methyl,methyl ester derivative of ochratoxin A.

The mycotoxin ochratoxin A (OA) was derivatized to an O-methyl,methyl ester (Me2) with diazomethane and then determined by high pressure liquid chromatography (HPLC). Both OA and OA-Me2 were chromatographed by reverse phase HPLC with a mobile phase of acetonitrile-water (60 + 40). An increase in retention time of 309 s was observed with OA-Me2 which was detectable at 254 nm at levels as low as 3 ng. Recovery of OA as OA-Me2 from chicken kidney homogenates and human plasma was quantitative following simple extraction and cleanup procedures, reaction with diazomethane, and HPLC analysis. The novel method described should prove useful for measuring and confirming OA in tissues and in further studies on the biological fate of this mycotoxin.

Animals↗

Inhibition of pancreatic carboxypeptidase A: A possible mechanism of interaction between penicillic acid and ochratoxin A.

Penicillic acid and ochratoxin A are environmentally important toxic fungal metabolites (mycotoxins) that are synergistic in combination. The effects of penicillic acid on the pancreatic enzyme, carboxypeptidase A were investigated in vitro and in vivo. A broad range of inhibition in vitro of the enzyme by PA was demonstrated with a half-maximal inhibitory concentration equal to 1.1 x 10(-4) M PA. Inhibition of carboxypeptidase A was time and temperature dependent, and resulted in decreased conversion of parent ochratoxin A to the non-toxic metabolite, ochratoxin alpha. Studies in vivo demonstrated a penicillic acid-dependent inhibition of pancreatic carboxypeptidase A activity in the mouse and the chicken following multiple oral exposure. It is postulated that the mode of toxic interaction of the two mycotoxins may be due, in part, to impaired detoxification of ochratoxin A through penicillic acid depletion of carboxypeptidase A activity.

Animals↗

A high pressure liquid chromatographic method for the analysis of zearalenone in chicken blood.

A high pressure liquid chromatographic (HPLC) method is described to determine zearalenone in chicken blood. Samples are extracted with acetonitrile, followed by a hexane cleanup procedure and extracted further with ethyl acetate. The analysis of zearalenone is by HPLC using a reverse phase radial compression separation system, an ultraviolet absorbance detector and a mobile phase of acetonitrile-water 60:40 (v/v). Recoveries of zearalenone in blood at levels of 50-200 ng/ml are in the range of 66.8-72.6%.

Animal Feed↗

Vanadium-induced inhibition of renal Na+, K+-adenosinetriphosphatase in the chicken after chronic dietary exposure.

Recent work has shown that V accumulates in the kidney and is a potent inhibitor of Na+, K+-adenosinetriphosphatase (ATPase) in vitro. Thus, as a nutritionally required element, V may regulate cation transport. The effect of chronic intake of the metal on Na+, K+-ATPase in vivo has not been reported. In this study laying strain chickens were fed calcium orthovanadate for 15 mo from d 1 of age at levels of 0, 25, 50, and 100 ppm in the diet. Whole tissue homogenates and 13,000 X g fractions were analyzed for ATPase activities. Concentrations of V producing 50% inhibition of Na+, K+-ATPase activity ranged from 1.0 X 10(-5) M in liver to 1.8 X 10(-6) M in kidney, which was the most sensitive tissue tested in vitro. Mg2+ -ATPase was more resistant to V than Na+, K+-ATPase. Studies in vivo suggested a V-dependent inhibition of renal Na+, K+-ATPase. Correlation of enzyme specific activity and levels of V in kidneys suggested V-ATPase mediated alteration in renal function.

Adenosine Triphosphatases↗

High pressure liquid chromatographic determination of aflatoxins by using radial compression separation.

A rapid method for the determination of aflatoxins was developed using high pressure liquid chromatography and a radial compression separation system. A standard solution of aflatoxins B1, B2, G1, G2, and M1 was analyzed solution of aflatoxins B1, B2, G1, G2, and M1 was analyzed at flow rates of 2.0 and 6.0 mL/min. Retention times, peak heights, and peak areas were reproducible over a 3-day period. Coefficients of variation for aflatoxin B1 at 2.0 and 6.0 mL/min were, respectively, 1.04 and 0.87% (retention time); 2.9 and 4.7% (peak height); and 8.2 and 4.7% (peak area). At 6.0 mL/min there was an approximate 25% loss in sensitivity but a greater than 50% reduction in retention time. Separation of all the aflatoxins was excellent using a dual flow rate of 2.0 mL/min with a change to 8.0 mL/min at 15 min post-injection. The applicability of the radial compression separation system for the rapid determination of aflatoxins in human tissues was also tested. Spiked samples of liver, serum, and urine-showed good resolution of all aflatoxin peaks at the higher flow rates.

Aflatoxins↗

Ochratoxin A and penicillic acid interaction in mice.

Penicillic Acid (PA) and Ochratoxin A (OA) are toxic fungal metabolites that are synergistic in combination. This interaction was investigated using mice which were doses orally as follows: control, none; solvent control, 0.2 ml bicarbonate buffer; PA, 40 mg/kg; OA, 10 mg/kg and combination, 40 mg/kg PA + 10 mg/kg OA. The only significant histopathologic change observed was an acute multifocal toxic tubular nephrosis which appeared most severe in the combination-treated mice killed on day 10. While the combination group had a death rate of 20% (5/25), no deaths occurred in the other treatment groups. The increased death rate and the extensive nephrotoxic findings in the combination group indicate a toxic interaction between OA and PA at sublethal dose levels and is consistent with a renal site of action.

Animals↗

High pressure liquid chromatographic determination of penicillic acid in chicken tissues.

Penicillic acid (PA) is a mycotoxin with reported cytotoxic, cardiotoxic, and carcinogenic activity and it can occur in high concentration in corn. The occurrence of PA in contaminated poultry feed represents a potential public health hazard. A reverse phase high pressure liquid chromatographic (HPLC) method is proposed for determining PA residues in chicken tissues. Optimization of chromatography was achieved for PA using a mobile phase consisting of acetonitrile: H2O. PA was detected by ultraviolet absorption at 254 nm, identified by retention time, and quantitated by peak area integration. Blood, parenchymal tissues, muscle, and alimentary tract contents were homogenized, sonicated, and acid treated followed by extraction with ethyl acetate and analysis by HPLC. Acute oral dosing of chickens with PA over a range of 50 to 550 mg/kg body weight resulted in detectable levels of the mycotoxin (confirmed by gas liquid chromatography) in gizzard muscle and contents, liver, kidney, heart, and intestinal contents. This method should prove useful both for the rapid and sensitive detection of PA residues in poultry and in further studies on the distribution and metabolism of this mycotoxin.

Animals↗

Effects of the aflatoxins on ATPase activities in mouse and rat liver.

The effects of the aflatoxins on ATPase activities in mouse and rat tissues were investigated in vitro. The hepatic oligomycin-sensitive (O.S.) Mg++ ATPase was inhibited significantly. The order of inhibition was G1 greater than B1 greater than G2 greater than B2. Mouse O.S. Mg++ ATPase was more sensitive than the corresponding rat enzyme. The oligomycin-insensitive (O.I.) Mg++ ATPase activities in rat and mouse liver were not altered. Although aflatoxins G1 and B1 were more potent inhibitors of hepatic O.S. Mg++ ATPase, no concentration-response was observed, whereas aflatoxins G2 and B2 inhibited enzyme activity in a concentration-dependent manner. Spectral analysis of aflatoxin G1 solutions suggested that solubility was not related to the observed effects. In addition, the effects of aflatoxin B1 and G1 on mouse brain microsomal Na+-K+ ATPase were examined. Although aflatoxin B1 was more potent that G1, both mycotoxins significantly inhibited enzyme activity in a concentration-dependent fashion.

Adenosine Triphosphatases↗

Inhibition of electrogenic sodium transport across toad urinary bladder by the mycotoxin patulin.

The effects of the mycotoxin patulin (4-hydroxy-4H-furo[3,2c]pyran-2(6H)-one) on short circuited intact toad bladder and on Na+-K+, activated ATPase were examined in an attempt to elucidate the relationship between toxin, the Na+-K+ ATPase enzyme system and associated active sodium transport. Patulin inhibited transbladder short circuit current and Na+-K+ ATPase from isolated bladder preprations. The effect was exponentially dependent on time. A significantly slower rate of inhibition was achieved within 15-30 min. The results are compatible with the assumption that Na+-K+ ATPase is associated with the pump mechanism since patulin inhibited enzyme activity and concomitantly reduced the rate of electrogenic Na+ transport. A significant correlation suggested a cause-effect relationship.

Animals↗

Aflatoxicosis in swine.

In an episode of aflatoxicosis in feeder pigs, mortality was about 20%. Histopathologic findings characteristic of experimentally induced aflatoxicosis and the finding of aflatoxin B1 in the serum of pigs and in the cornbased feed confirmed the diagnosis. Aflatoxins B1 and B2 were found in the corn used to prepare the feed. Combine harvesting of the corn, which cracked a large percentage of the kernels, coupled with prolonged drying time of the corn probably contributed to the aflatoxin production. Although the corn was fed to adult swine without observable effect, 47 of the 250 feeder pigs developed typical signs of aflatoxicosis. Unseasonably cold weather apparently was a factor in initiating the onset of clinical signs and probably increased the severity of the disease.

Aflatoxins↗

Effects of rubratoxin B on the kinetics of cationic and substrate activation of (Na+-K+)-ATPase and p-nitrophenyl phosphatase.

Rubratoxin B, a lactone-containing bisanhydride metabolite of certain toxigenic molds, inhibited (Na+-K+)-stimulated ATPase activity of mouse brain microsomes in a dose-dependent manner with an estimated IC50 of 6 x 10(-6) M. Hydrolysis of ATP was linear with time and enzyme concentration, with or without rubratoxin in reaction mixtures. Altered pH and activity curves for (Na+-K+)-ATPase demonstrated comparable inhibition by rubratoxin in buffered acidic, neutral, and alkaline pH ranges. Kinetic studies of cationic-substrate activation of (Na+-K+)-ATPase indicated classical competitive inhibition for Na+ and K+. Results also showed competitive inhibition for K+ activated p-nitrophenyl phosphatase as demonstrated by altered binding site parameters without change in the catalytic velocity of dephosphorylation of the enzyme . phosphoryl complex. Noncompetitive inhibition with regards to activation by ATP and p-nitrophenyl phosphate was indicated by altered Vmax values with no change in Km values. Inhibition was partially restored by repeated washings. Preincubation with sulfhydryl agents protected the enzyme from inhibition. Cumulative inhibition studies with rubratoxin and ouabain indicated possible interaction between the two inhibitors of (Na+-K+)-ATPase. Rubratoxin appeared to exert its effects on (Na+-K+)-ATPase by interacting at Na+ and K+ sites.

4-Nitrophenylphosphatase↗