Diphenylhydantoin Treatment Research Articles

Effects of diphenylhydantoin on locomotion and thigmotaxis of larval zebrafish

Antiepileptic effects of diphenylhydantoin (DPH) have been documented in animal studies and clinical research, while little is known about the effects of the drug on basic behaviors and anxiety-related behaviors. In order to understand neuroactivities of DPH deeply and administrate DPH in clinic rationally, it is necessary to study neurobehavioral effects of the drug. In the present study, the effects of DPH on the locomotor activity and thigmotaxis of zebrafish larvae at 5days post fertilization (dpf) were explored under different illumination conditions. The influence of DPH on zebrafish larval responses to visual stimuli (sudden illumination transition from light to dark) was also investigated. Under light or dark condition, exposure to high concentrations of DPH resulted in decreased locomotor activity and thigmotaxis, whereas DPH treatment at low doses enhanced the locomotor activity. Additionally, sudden illumination transition induced robust increase in the locomotor activity and this phenomenon was not modified by DPH treatment. Our results suggest that DPH has potential stimulatory and inhibitory effects on the locomotor activity and possesses anxiolytic properties. In addition, responses of 5-dpf zebrafish larvae to visual stimuli were not modified by DPH treatment.

Diphenylhydantoin plays a role in gene expression related to cytoskeleton and protein adhesion in human normal palate fibroblasts

Morphogenetic processes during palate development are related to extracellular matrix composition. The cell-extracellular matrix relation plays a role in cell activity and in gene expression. We studied the effect of diphenylhydantoin, a teratogen known to induce cleft palate in human newborns, on extracellular matrix production. We investigated whether diphenylhydantoin treatment caused any differences in glycosaminoglycans, collagen synthesis and gene expression in human normal palate fibroblasts. Human palate fibroblasts were maintained for 24 hours in serum-free 199 medium containing 5 microg/mL (3)H-glucosamine or (3)H proline hydrochloride. Collagen and glycosaminoglycan classes were then measured using biochemical methods, gene expression with microarray analysis and cytoskeleton components with immunofluorescent antibodies and computer analysis. In normal fibroblasts diphenylhydantoin reduced collagen and glycosaminoglycan synthesis with a marked effect on sulphated glycosaminoglycans. There were also substantial decreases in tubulin, vimentin and alpha-actin staining and an increase of vinculin compared to controls. Diphenylhydantoin acted on several genes related to the synthesis of cytoskeleton and adhesion membrane proteins. It inhibited caderin, caveolin, RTK and alpha-actin, and increased nectin, cytoplasmatic FRG vinculin, ITGA, ITGB extracellular matrix ligand and EDG2 gene expression. DNA binding gene expression, which plays a role in cell growth and senescence, was activated. Since cell activity is dependent on the cell morphology and extracellular matrix composition, these findings indicate that in human normal palate fibroblasts diphenylhydantoin can modify cytoskeletal components and extracellular matrix-cell adhesion, with consequent effects on gene expression. These changes might be related to anomalous palate development.

Cytomegalovirus antibodies in epileptics receiving diphenylhydantoin

Cytomegalovirus (CMV) antibodies were determined by indirect haemagglutination in 53 epileptics receiving long-term diphenylhydantoin (DPH) therapy and in 53 matched controls. Absorption of serum IgG, IgA and IgM performed in 12 sera showed that the antibodies were of the IgG class. A decreased incidence of high CMV antibody titres (greater than or equal to 320) was found in epileptics (11.3%) compared with controls (34.0%) (0.02 greater than P greater than 0.01), whereas antibody titres greater than or equal to 40 were found in 37.7% of epileptics and in 45.3% of controls (n.s.). The CMV antibody incidence and titre range were similar in patients with symptomatic (37.9%) and idiopathic (37.5%) epilepsy, suggesting that the DPH treatment was responsible for the decreased antibody occurrence in patients. No correlation between CMV antibody titres and the serum immunoglobulin levels or the DPH concentration or clearance could be established in the epileptics.

Effects of withdrawal from long-term diphenylhydantoin treatment on audiogenic and maximal electroshock-induced seizures in rats.

Rats withdrawal from long-term diphenylhydantoin treatment (DPH) were tested for their sensibility to convulsant stimuli. Animals were more sensitive to convulsions elicited by maximal electroshock and sound, respectively, at 48 and 72 h after drug removal. These results suggest that long-term DPH treatment might develop a central nervous system supersensitivity state.

Effects of prenatal diphenylhydantoin treatment on reproductive outcome, development, and behavior in rats

Pregnant Sprague-Dawley rats were administered sodium diphenylhydantoin (DPH) by gavage on gestation days 9–18 in doses of 0, 100, or 200 mg/kg. DPH-treated dams showed a dose-related decrease in weight gain throughout the drug treatment period. Offspring had significantly lowered birth weights, along with increased mortality through the first 30 days of life. In one of the experiments specifically designed to measure pup mortality, the rates were 41.4% and 60.7% respectively, for the 100 and 200 mg/kg DPH groups. Shortly after eye opening, many of the DPH-treated animals developed chromodacryorrhea, a condition that remained throughout the experiment. Aside from lowered body weights, no differences were observed in the attainment of several standard developmental landmarks. During the neonatal period, the pups exposed in utero to 100 mg/kg DPH showed a significant increase in pivoting locomotion on postnatal days 7 and 9. As adults, the animals exposed to 200 mg/kg DPH showed significant increases in locomotor activity measures. Both DPH groups developed a dose-related and highly abnormal spontaneous circling behavior. These results confirm a small but growing body of literature that demonstrates that DPH is a behavioral teratogen.

A case of amyloidosis associated with diphenylhydantoin therapy.

Prolonged administration of diphenylhydantoin (DPH) has been implicated as a possible etiologic factor in immunological aberrations and lymphoproliferative disorders. Diphenylhydantoin may account for the increase in susceptibility to lymphoproliferative diseases, as a result of its immunosuppressive effect. We report a case of amyloidosis with monoclonal gammopathy which developed during DPH treatment, without multiple myeloma or lymphoproliferative disorders. The association between DPH and monoclonal gammopathy is very rare, and such a case of amyloidosis associated with DPH has not been reported previously. DPH, however, may have played a role in the development of monoclonal gammopathy, which was the precursor of amyloid protein.

The influence of long-term anticonvulsant therapy with diphenylhydantoin and carbamazepine on serum gamma-glutamyltransferase, aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase.

In 110 patients receiving long-term anti-convulsant monotherapy with diphenylhydantoin (DPH) and carbamazepine (CBZ) the serum activities of gamma-glutamyltransferase (gamma-GT), aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase (AP) were examined retrospectively. Elevated serum levels of gamma-GT and AP were seen in 91% and 39% of patients receiving DPH therapy compared to 64% and 14% of those receiving CBZ treatment. With all enzymes evaluated increases were more frequent and higher with DPH treatment than with CBZ. Frequency and extent of increased activity of gamma-GT were highly related to daily dosage in both preparations. The proportion of pathological enzyme levels was associated with age in DPH and CBZ therapies but not found to be significant. Sex differences in the frequency of increased enzyme activities could not be demonstrated. The results are discussed in the context of induction of the cytochrome P-450 system.

Comparison of basal glutathione S-transferase activities and of the influence of phenobarbital, butylated hydroxy-anisole or 5,5'-diphenylhydantoin on enzyme activity in male rodents

1. Hepatic cytosolic glutathione S-transferase (GST) activities, toward five substrates, were shown to vary markedly among three laboratory rodent species. 2. Basal GST activities for 1-chloro-2,4-dinitrobenzene (hamster > mouse > rat), 1,2-dichloro-4-nitrobenzene (mouse > rat > hamster), p-nitrobenzyl chloride (rat = mouse = hamster), bromosulfophthalein (rat > mouse > hamster) and 1,2-epoxy-3-( p-nitrophenoxy)propane (mouse > rat = hamster) differed with respect to magnitude and distribution among species. 3. GST substrate activities in response to phenobarbital, butylated hydroxy-anisole or 5,5'-diphenylhydantoin treatment were increased more often in mouse and rat as compared to the hamster. 4. These results suggest that basal GST activity, as well as inducibility, differ among rodent species. Since GST are involved in detoxication processes, differences in GST properties may underlie variability in species sensitivity to toxicants.

Induction of a Novel Ca 2+-Dependent Chymotrypsin-Like Serine Protease by Tumor Promoters in Rat Livers<xref ref-type="fn" rid="FN1">2</xref>

Induction of a microsomal Ca2+-dependent serine protease by hepatic tumor promoters was studied. Male F344 rats were fed a diet containing one of the following promoting agents: phenobarbital (CAS: 50-06-6), dichlorophenyltrichloroethane (CAS: 50-29-3), butylated hydroxytoluene, ethyl-alpha-chlorophenoxyisobutyrate (CAS: 128-95-0), or 17-alpha-ethynylestradiol (CAS: 57-63-6) or a nonpromoting agent, diphenylhydantoin (CAS: 57-41-0), for 1 week. By treatment with promoters, the protease activity in the microsomal fraction was increased to threefold to fivefold that of control, whereas only a slight increase of activity was found after diphenylhydantoin treatment. The Ca2+-dependent protease activity was determined with the use of N-benzoyl-L-tyrosine ethyl ester as the substrate in a medium containing 50 mM CaCl2 for its maximal activity. This protease was preferentially localized in the smooth microsomal membrane and strongly inhibited by diisopropyl phosphorofluoridate (CAS: 55-91-4), and the optimum pH of the activity was 7.8. It appears that the Ca2+-dependent serine protease measured by using a chymotrypsin substrate is a novel protease, and induction of its activity by hepatic tumor-promoting agents is a common and specific phenomenon.

Dopaminomimetic action of diphenylhydantoin in rat striatum: effect on homovanillic acid and cyclic AMP levels.

Diphenylhydantoin (DPH) is known to induce reversible paroxysmal dyskinesias and paranoid psychosis in humans, but its interactions with dopamine (DA) metabolism are not clear. Single doses of DPH (60-100 mg kg-1), with serum levels over 10 micrograms ml-1, reduced homovanillic acid (HVA) levels in rat striatum. The DPH-induced HVA decrease was enhanced by supersensitivity of postsynaptic DA receptors following chronic haloperidol (Hal) administration. DPH 60 mg kg-1 given acutely enhanced the HVA decrease induced by apomorphine (Apo) and partially counteracted the HVA increase following acute Hal (0.1-0.5-2 mg kg-1). After chronic DPH treatment, Apo was ineffective in reducing striatal HVA levels. Concomitant chronic treatment with DPH and Hal counteracted the development of supersensitivity of postsynaptic DA receptors to Apo. Single doses of DPH (30-60-100 mg kg-1) increased cyclic AMP striatal content; this effect was blocked by Hal. A dopaminomimetic DPH action and a subsensitivity of postsynaptic DA receptors after chronic DPH seem to be suggested. These effects could be related to the dyskinetic and psychotic syndromes produced by the drug.

Collagen metabolism in two rare forms of epidermolysis bullosa.

The collagenase activity in skin fibroblast cultures from three patients with rare forms of epidermolysis bullosa was assayed before and after proteolytic activation of the medium. Two of the patients had the recessive dystrophic form of the disease (REBD), which is generally associated with abnormal collagenase activity. The other patient had an atrophic mitis form of the disease (REBA), which has not previously been associated with defective collagen metabolism. However, a similar increase in collagenase activity was found in all three cases. The total collagen production of EB fibroblasts was also enhanced, being two to five times that of control cell lines, and the intracellular hydroxylases of collagen biosynthesis were higher in the case of two EB-cell lines. These changes reflect the compensatory increase in collagen synthesis which follows the increased degradation caused by excessive free collagenase activity. Diphenylhydantoin treatment of one REBD patient for 9 months improved her condition.

Diphenylhydantoin inhibits the thyrotropin response to thyrotropin-releasing hormone in man and rat.

We studied the effect of diphenylhydantoin (DPH) on TRH-stimulated TSH secretion in four patients (three of whom had hypothyroidism; one treated with L-T4), in euthyroid rats, and in dispersed rat anterior pituitary cells in short term tissue culture. DPH treatment of four patients (100 mg, three times daily for 14 days) caused a 22% decrease in mean serum T4 (P < 0.06), a 31% decrease in mean serum free T4 (P < 0.01), and a 6% decrease in mean serum T3. Mean basal serum TSH was unchanged, but the integrated TSH response after TRH injection (500 μg, iv) decreased 28% (P < 0.06) and 51% (P < 0.01) after 7 and 14 days of DPH treatment, respectively. Changes in the integrated PRL response to TRH did not correlate with those in TSH.Injection of DPH into euthyroid rats (5 mg/100 g BW·day for 9 days) caused a significant decrease in mean serum T4 but no change in TSH release after TRH treatment. However, 1 h after a large dose of DPH (20 mg/100 g BW), there was a marked reduction (58%) in TRH-induced TSH releas...

Effect of 5,5'-diphenylhydantoin on the activities of hepatic cytosol malic enzyme and mitochondrial alpha-glycerophosphate dehydrogenase in athyreotic rats.

Since 5,5'-diphenylhydantoin (DPH) decreases specific nuclear receptor binding of T3, the possibility that DPH might affect thyroid activity was examined by measurement of the activity of rat hepatic cytosol malic enzyme (ME) and mitochondrial alpha-glycerophosphate dehydrogenase (alpha-GPD) after DPH administration. Groups of athyreotic rats, prepared by thyroidectomy and 131I administration, were injected with vehicle (V), DPH, T3, or a combined dose of DPH and T3 for 48 h. The rats were then killed, and liver fractions were prepared for the measurement of ME and alpha-GPD activities. Compared with the V group, DPH treatment resulted in a significant increase in mean ME activity in four experiments; the mean ME activity for all studies was 2.2-fold greater than that of the V group (P less than 0.001). A significant increase in mean alpha-GPD activity was not observed in DPH-treated rats. T3 treatment resulted in a mean 31.5-fold increase in ME activity and a 10.9-fold increase in alpha-GPD activity. The magnitude of T3 induction of both enzymes was substantially attenuated by simultaneous injection of DPH, to 54-66% of the activity induced by T3 alone (P less than 0.001). Pair-feeding a T3-injected group and the DPH plus T3 group did not result in a decrease in the activity of either enzyme. Moreover, the observed changes did not appear to result from alterations in enzyme determinations or from significant changes in cytosol or mitochondrial protein concentration, cytosol T3, or plasma glucose. Although DPH-induced changes in the fractional rate of enzyme metabolism cannot be excluded, it is possible that changes in enzyme activities observed after DPH injection resulted from an interaction between DPH and the nuclear T3 receptor. If so, based on the effects observed in the present studies, DPH might be classified as a partial thyroid agonist.

CHRONIC DIPHENYLHYDANTOIN THERAPY DOES NOT REDUCE PLASMA 25‐HYDROXY‐VITAMIN D

A randomized controlled study was performed to investigate the effect of 2 years' monitored diphenylhydantoin (DPH) therapy on plasma 25-hydroxyvitamin D (25-OHD) in non-epileptic, non-institutionalized subjects. Mean +/- SEM plasma 25-OHD of 18 DPH-treated subjects at the end of 2 years' drug treatment was 59 +/- 8 nmol/l (23.6 +/- 3.2 ng/ml), which was not decreased compared to that of eighteen control subjects (54 +/- 8 nmol/l, 21.6 +/- 3.2 ng/ml). In addition, mean plasma 25-OHD had not changed 1 month after ceasing DPH. The treated group had a higher mean serum alkaline phosphatase (SAP) during DPH treatment, attributable to hepatic enzyme induction. It is concluded that therapeutic doses of DPH without other anticonvulsants do not have a clinically significant effect on plasma 25-OHD.

Transient hemiparesis: A rare manifestation of diphenylhydantoin toxicity

Among the common side effects of diphenylhydantoin (DPH) overdose, the most frequently encountered neurological signs are those of cerebellar dysfunction. Very rarely, the toxic neurological manifestations of this drug are of cerebral origin. Two patients are presented who suffered progressive hemiparesis due to DPH overdose. Both had brain surgery before DPH treatment. It is assumed that patients with some cerebral damage are liable to manifest DPH toxicity as focal neurological signs.

Hepatic drug metabolism after phenobarbital and diphenylhydantoin administration in the rat—influence of vitamin D 3 status

Measurements of aniline hydroxylation, aminopyrine N-demethylation and cytochrome P-450 content after a 3-week treatment with phenobarbital (PB), diphenylhydantoin (DPH) or a combination of the two drugs were undertaken during normal vitamin D status (D +) and vitamin D deficiency (D −) with or without vitamin D 3(D 3) supplementation. Serum calcium concentrations were reduced after D deprivation but responded by a significant increase toward normal values to a single pharmacological dose of D 3. Serum phosphorus concentrations were also slightly raised by the supplementation. Even in the presence of higher cytochrome P-450 content in D − rats, aniline hydroxylase and aminopyrine N-demethylase activities were lower in D − than in D + animals. These two enzymatic parameters, as well as cytochrome P-450 content, were increased by anticonvulsant (ACV) drug treatment regardless of the D nutritional status. The in vivo hexobarbital sleeping time was shortened by ACV drugs but the sleeping time tended to be longer in D − than in D + rats. Supplementation with 1000 I.U. of D 3, lowered aniline hydroxylase activity both in D + and D− animals; the supplementation had no effect on aminopyrine N-demethylase activity in D + animals but had an inhibitory effect after PB and a stimulatory effect after DPH treatment in D− animals. Cholecalciferol supplementation lowered cytochrome P-450 content toward normal values in D− rats while it had no effect in D + animals. These observations suggest that (1) PB and DPH pretreatment do not alter the normal response of serum calcium and phosphorus to a single pharmacological dose of D 3; (2) in a state of vitamin D deficiency accompanied by hypocalcemia, the inducing capacity of PB and DPH on the liver mixed function oxidase system is not lost; (3) under certain circumstances, vitamin D 3, can influence the catalytic activity of the mono-oxygenase complex; (4) cytochrome P-450 is influenced by vitamin D deficiency and/or changes in extracellular calcium but the forms induced by PB and DPH may not necessarily be the ones specifically involved in vitamin D metabolism.

Effect of diphenylhydantoin on patterns of insulin secretion in obese subjects.

The effect of short-term treatment with diphenylhydantoin (DPH) on the insulin secretion patterns during OGTT and on the daily insulin profile was studied in obese patients. DPH treatment for 3 days with a dose of 300 mg/die (100 mg, 3 times daily) significantly decreased the insulin release after glucose ingestion, but did not alter the basal insulin level. No effect on the fasting glucose concentration as well as on the glucose profiles during OGTT was observed after short-term DPH treatment. A smaller decrease of plasma free fatty acid concentration during OGTT performed after DPH administration confirmed the inhibitory effect of the drug on insulin release. Short-term DPH treatment was also shown to decrease markedly the postpradial insulin release in obese patients. No difference was noted between plasma 11-OHCS and serum HGH concentrations during OGTT before and after DPH treatment. The possible therapeutic role of DPH in obesity is discussed.

DIFFERING POLYAMINE RESPONSES DURING TERATOGENESIS WITH DIPHENYLHYDANTOIN AND FOLIC ACID ANTAGONIST

Polyamine content is correlated with nucleic acid synthesis in rapidly growing tissue and may serve as an indicator of damage by toxins, including teratogens. This report concerns the alterations in embryonic polyamine content after exposure to teratogens. Primagravid Swiss albino mice were injected intraperitoneally on day 9 with diphenylhydantoin (DPH), 88 mg/kg body weight; embryos were recovered on day 11. Pregnant Long-Evans rats were intubated on day 10 and a modified diet was supplemented with 9-methyl pteroylglutamic acid (9-methyl PGA) on days 10-13. Spermidine, spermine and putrescine were quantitated on a Durrum D-500 amino acid analyzer, and are reported as nmoles/mg of embryonic protein. Rat embryos from pregnancies treated with 9-methyl PGA had unaltered polyamines. Mouse embryos whose mothers were exposed to DPH had significant reductions in spermidine and spermine, but putrescine levels remained comparable. Growth during polyamine accumulation is attributed to their stimulation of dihydrofolate reductase, the enzyme antagonized by 9-methyl PGA. Since polyamines were unchanged during this antagonism, they may play no direct role in the reactivation of this enzyme system. Following DPH treatment embryonic DNA was lowered proportionally more than protein content, suggesting a block in DNA synthesis. Folate deficiency has been suggested as the mechanism of DPH teratogenesis. However, the differing polyamine responses to the 9-methyl PGA and DPH teratogenic regimens indicate different mechanisms may exist.

Assessment of diabetogenic drug activity in the rat: diphenylhydantoin.

An animal model is described that can be used to detect drugs that may exacerbate or ameliorate diabetes. The design of this model is based upon the finding that hyperglycemia caused by intravenous administration of streptozotocin to rats is inversely correlated with pancreatic insulin concentration when expressed as total pancreatic insulin (ng) per body weight (g). Drugs that alter this relationship may be classified, in a preliminary manner, as ameliorative or diabetogenic. The diabetogenic activity of diphenylhydantoin (DPH) via oral administration was assessed in both normal and streptozotocin diabetic rats. Rats were fed powdered chow diet, with or without 0.2% (w/w) DPH, for 19 days. Food consumption and rat weight were recorded daily; whole blood glucose concentrations were determined at the start of the study and at the midpoint. At sacrifice liver and pancreas were excised and blood samples were collected. Protein, glycogen, and lipid levels were determined in liver; insulin in pancreas; and insulin, ketone bodies, glucose, and Iipid in blood. DPH treatment did not affect growth or food consumption. The drug dose, calculated from the food consumption data, was 139 mg/kg‐day for the normal rats and 311 mg/kg‐day for the diabetic rats. DPH increased liver weight and lipid content in both normal and diabetic rats and lowered blood serum triglyceride concentration in normal rats. However, the concentrations of whole blood glucose, blood serum insulin, and pancreatic insulin were not altered by DPH treatment. The results indicate that the diabetogenic side effect of DPH cannot be observed in rats when the drug is administered orally.

The effect of diphenylhydantoin on thyroid function.

ABSTRACT The effect of diphenylhydantoin (DPH) on serum T4, a free T4 index, serum T3, serum TSH, TSH response to iv TRH and absolute iodine uptake (AIU) of the thyroid gland was studied in 26 patients receiving long-term DPH treatment and was compared to controls. In the DPH-treated group, serum T4, free T4 index and serum T3 were decreased to 75% of control values. Serum TSH and AIU were significantly increased. Twelve patients were studied before and after 1–2 weeks of treatment with DPH and a significant decrease in serum T4 and free T4 index was noted. Serum T3 decreased but not significantly. Serum TSH was significantly increased, however, the increase in TSH response to TRH and in AIU were not statistically significant. In 7 patients with hyperthyroidism 2–4 weeks' treatment with DPH was followed by a significant decrease in serum T4, serum T3 and free T44 index. It is suggested that DPH primarily acts as a stimulator of hepatic degradation of the thyroid hormones.