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Biomedical subjects

H L Kim

Publications and source records attributed to H L Kim.

At least 73 records · Page 4Linked to original sources

Experimental prevention of bitterweed (Hymenoxys odorata) poisoning of sheep.

To examine the effects on bitterweed toxicity of dietary factors known to increase thiol concentrations in the body, 36 lambs were fed one of the following diets (12 lambs/diet) for a minimum of 9 days prior to bitterweed administration: diet 1, 10% crude protein; diet 2, 20% crude protein, 0.5% methionine, 0.5% sodium sulfate, and 1,102 IU of vitamin E/kg; and diet 3, diet 2 with 0.5% ethoxyquin hydrochloride added. Four lambs fed each diet were euthanatized prior to bitterweed administration (initial euthanasia group). Four lambs fed each diet were administered bitterweed (0.68% hymenoxon, air-dried basis) at a rate of 0.25% of live weight for 5 consecutive days. The remaining four lambs on each diet served as unchallenged controls. In the initial euthanasia group, diet 2 increased extracellular blood thiol concentrations (1.12 vs 0.94 mg of SH/d1, P less than 0.10), rumen fluid thiol concentrations (4.46 vs 1.88 mg of SH/d1, P less than 0.05), and liver thiol concentrations (263.6 vs 109.3 micrograms SH/g of wet wt, P less than 0.05), compared with diet 1. Ethoxyquin hydrochloride (diet 3) reduced blood thiol concentrations (0.94 vs 1.12 mg of SH/dl, P less than 0.10) and liver thiol concentrations (151.6 vs 263.6 micrograms of SH/g of wet wt, P less than 0.05), compared with diet 2. Kidney thiols were unaffected by treatments.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗

Constitutive and Aroclor 1254-induced hepatic glutathione S-transferase, peroxidase and reductase activities in genetically inbred mice.

1. Constitutive and Aroclor 1254-induced hepatic glutathione (GSH) S-transferases, GSH peroxidase and GSH reductase activities were determined in 12 strains of 8-10 week-old inbred male mice. 2. The constitutive GSH S-transferase activity varied from 2.5 (SJL/JCR) to 8.9 (C57BL/6N) mumol/min/mg protein and the corresponding values for the Aroclor 1254-treated mice were in the range of 7.1-23.0 mumol/min/mg protein. Aroclor 1254 significantly induced GSH S-transferase activity in all mice, however, significant interstrain differences were found in inducibility. 3. Aroclor 1254-treatment caused a 4.2-fold induction of GSH S-transferase in NFS/NCR but only a 1.4-fold increase in AKR/NCR mice. Aroclor 1254 significantly induced GSH reductase in all strains studied while GSH peroxidase activity decreased in these mice. 4. The range of hepatic GSH levels in control and Aroclor 1254-treated mice was relatively narrow for both groups (6.59-11.25 microM/g wet tissue).

Animals↗

Role of glutathione in the toxicity of the sesquiterpene lactones hymenoxon and helenalin.

Hymenoxon and helenalin are toxic sesquiterpene lactones present in the toxic range plants Hymenoxys odorata and Helenium microcephalum. Helenalin (25 mg/kg) or hymenoxon (30 mg/kg) administered to immature male ICR mice caused a rapid decrease in hepatic glutathione levels and were lethally toxic to greater than 60% of the animals within 6 d. L-2-Oxothiazolidine 4-carboxylate (OTC), a compound that elevates cellular glutathione levels, administered to mice 6 or 12 h before either helenalin or hymenoxon protected against hepatic glutathione depletion and the lethal toxicity of these toxins. OTC administered at the same time as the sesquiterpene lactones was not protective, suggesting that the critical events against which glutathione is protective occur within the first 6 h. In primary rat hepatocyte cultures, hymenoxon and helenalin (4-16 microM) caused a rapid lethal injury as determined by the release of lactate dehydrogenase. Cotreatment of cultures with N-acetylcysteine at high concentrations (4 mM) afforded significant protection against lethal injury by both toxins. In contrast, BCNU, which inhibits glutathione reductase, or diethylmaleate, which depletes hepatocellular glutathione, potentiated the hepatotoxicity of helenalin and hymenoxon in monolayer rat hepatocytes. These studies suggest that the in vivo and in vitro toxicity of hymenoxon and helenalin is strongly dependent on hepatic glutathione levels, which hymenoxon and helenalin rapidly deplete at very low concentrations.

Acetylcysteine↗

The sesquiterpene lactone hymenoxon acts as a bifunctional alkylating agent.

Hymenoxon, a toxic sesquiterpene lactone found in bitterweed, bound deoxyguanosine in a cell-free system and formed adducts with guanine residues in cellular DNA. The reactive dialdehyde form of hymenoxon formed stable Schiff base products with deoxyguanosine which were separable from unreacted hymenoxon and deoxynucleosides by reverse-phase high pressure liquid chromatography. Hymenoxon adducts which eluted as a single impure peak from the octadecylsilane column separated on amino and diphenyl-bonded phases with 10% methanol. Tritiated nucleoside adducts were isolated and purified from CFW mouse sarcoma cells treated with hymenoxon. Proton nuclear magnetic resonance spectra of purified hymenoxon-deoxyguanosine adducts revealed a loss of signals for hydroxyl groups in the bishemiacetal of hymenoxon. 13C-nuclear magnetic resonance spectra revealed that the major adduct has 35 carbon atoms, indicating an interaction of at least two guanine residues per hymenoxon molecule and suggesting that hymenoxon may cross-link DNA. Sedimentation analysis of treated DNA further showed that DNA cross-linking by hymenoxon (30 micrograms/ml) was equivalent to that of a known cross-linking agent, mitomycin C (7.5 micrograms/ml). Hymenoxon was more cytotoxic to DNA cross-link repair-deficient Chinese hamster ovary cell mutants than to repair-proficient strains. These data combine to indicate that hymenoxon acts as a bifunctional alkylating agent which cross-links DNA in mammalian cells.

Alkylating Agents↗

Depletion and resynthesis of tissue thiols by helenalin and tenulin.

The depletion of tissue thiols by two anti-leukemic agents helenalin and tenulin was studied in mice. Helenalin (100 mg/kg) and tenulin (213 mg/kg) in dimethyl sulfoxide (DMSO) were administered orally, and the acid-soluble total thiol concentrations in the liver, kidney, stomach and small intestine were determined at 0.5, 1.0, 3.0, 6.0, 12.0 and 24.0 hr post-treatment. The lowest tissue thiol levels (42-84% of pretrial values) were observed within 0.5 - 1.0 hr following helenalin treatment. The tenulin treated mice exhibited the lowest values (46-80% of pretrial control values) at 3.0 hr post-treatment in all tissues except in the stomach, which reached its lowest value at 1.0 hr. The most dramatic effects were found in the stomach and small intestine where the mean thiol concentrations varied from 43% to 141% (at 1.0 and 24.0 hr) and from 76% to 169% (at 0.5 and 24.0 hr) of pretrial values, respectively, among helenalin treated mice. The tissue thiol levels in DMSO- and physiological saline (PSS)-treated mice fluctuated probably due to fasting and/or diurnal changes.

Animals↗

Increased numbers of neurons occur in the inferior colliculus of the young genetically epilepsy-prone rat.

To determine if the increase in the number of neurons observed in the inferior colliculus (IC) of the adult genetically epilepsy-prone rat (GEPR) as compared to the Sprague-Dawley rat was present in the young GEPRs prior to the time at which seizure activity commences, brains from both types of rats 4-10 days of age were studied. A statistically significant increase in the numbers of small neurons occurred in the IC of the young GEPR. At 4 days of age, a 55% increase in the number of small neurons was found in the GEPR as compared to the Sprague-Dawley rat and at 10 days of age this increase was 105%. The numbers of the medium and large neurons were similar in the older group of rats. These data suggest that the increase in cell number observed in the adult GEPR is not compensatory to the seizure activity, but is genetically programmed.

Acoustic Stimulation↗

Alkylation of deoxyguanosine by the sesquiterpene lactone hymenoxon.

Hymenoxon, a toxic sesquiterpene lactone found in the ruminant forage plant Hymenoxys odorata, binds deoxyguanosine in a cell-free system, and forms adducted guanine residues in sheep lymphocyte DNA. Mitogen-stimulated DNA synthesis in lymphocytes was inhibited by hymenoxon at concentrations greater than 100 microM. Unscheduled DNA synthesis in lymphocytes was initiated by hymenoxon concentrations exceeding 50 microM, and inhibited by concentrations above 100 microM. We describe an HPLC method which separates unmodified hymenoxon and deoxyguanosine from the hymenoxon-deoxyguanosine adduct, and allows the quantitation of adducts in hymenoxon-treated cells.

Alkylation↗

Preparation and dietary effects of ethoxyquin hydrochloride.

Ethoxyquin (EQ, 1,2-dihydro-6-ethoxy-2,2,4-trimethylquinoline) was purified and converted to a crystalline, stable ethoxyquin hydrochloride (EQ-HCl). The readily available (technical grade) oily EQ reacted with concentrated hydrochloric acid (HCl) and precipitated as a crystalline salt (EQ-HCl) in acetone, leaving most of the impurities in solution. The regenerated free base (EQ) from the EQ-HCl was further purified by silicagel column chromatography to remove several minor contaminants, and the pure unstable EQ was immediately converted into a pure stable salt (EQ-HCl). The dietary administration of EQ-HCl, 0.25 or 0.5% in the feed, induced hepatic and intestinal thiols in mice and provided protection against toxic doses of pyrrolizidine alkaloids. The LD50 values of the 0.125 and 0.25% EQ-HCl-pretreated mice were 94.0 and 98.5 mg/kg, respectively, compared to that of controls, 71.3 mg/kg. The EQ-HCl-supplemented feed appeared to be more palatable, but other effects, such as the hepatic hypertrophy, the tissue thiol induction, and the protective effects, were comparable to those of unpurified EQ.

Animals↗

Effects of SKF 525-A, phenobarbital and 3-methylcholanthrene on the toxicity of lobeline sulfate.

Pretreatment of mice with the microsomal enzyme inducers and inhibitors modified the toxicity of lobeline sulfate. The intraperitoneal LD50 of lobeline sulfate following SKF 525-A (75 mg/kg), phenobarbital (PB), and 3-methylcholanthrene (3-MC) were 18.3, 85.5 and 82.1 mg/kg, respectively, as compared to that of saline treated controls, 55.3 mg/kg. Pretreatment of mice with disulfiram (DSF), diethylmaleate (DEM), or ethoxyquin hydrochloride (EQ-HC1) exhibited a very slight effect on the toxicity of lobeline sulfate. These results suggest that the hepatic microsomal monooxygenase system, but not glutathione (GSH), is involved in the detoxication of lobeline sulfate.

Animals↗

Tissue thiol induction in mice and protective effect against pyrrolizidine alkaloids by dietary ethoxyquin and methionine hydroxy analog.

Dietary administration of ethoxyquin (EQ) and methionine hydroxy analog (MHA) induced the thiol concentrations in liver, kidney, small intestine and stomach of mice accompanied with hepatic hypertrophy. The hepatic thiol levels among EQ, EQ-MHA, or EQ-methionine treated mice were significantly higher (p less than 0.01) than that of the controls. Hepatic hypertrophy was apparent with 0.031% EQ-0.063% MHA in the diet. EQ feeding, as 0.5% w/w of feed with or without another additive, also resulted in reduced weight gain. The toxicity of pyrrolizidine alkaloids (PAs) was reduced by feeding diets containing 0.063% EQ-0.125% MHA or 0.125% EQ-0.25% MHA. The intraperitoneal LD50 values of PAs in these two EQ-MHA feeding groups were 92.8 and 94.0 mg/kg, respectively, compared to that of 71.3 mg/kg for the controls.

Animals↗

Elimination of adverse effects of ethoxyquin (EQ) by methionine hydroxy analog (MHA). Protective effects of EQ and MHA for bitterweed poisoning in sheep.

Dietary ethoxyquin (EQ) and methionine hydroxy analog (MHA) protected 6-8-month-old wethers from toxic doses of bitterweed (Hymenoxys odorata DC.). The EQ-MHA group received sweet feed (corn, oats, dehydrated alfalfa pellets, cane molasses and minerals), 500 g/day/sheep, supplemented with EQ and MHA (0.5% and 1.0% of feed, respectively) for 9 days prior to the poisoning with bitterweed while the MHA group received the same feed without EQ and controls received the same amount of feed with no additives. Two of 6 MHA-treated and 3 of 7 controls died whereas all 7 EQ-MHA-treated sheep survived after receiving 5 doses of bitterweed (5 X 5.5 g/kg) in 6 days. Coadministration of MHA and EQ eliminated the adverse effect of EQ; dietary EQ lowered the serum albumin, calcium, and alkaline phosphatase content while protecting the animals from bitterweed poisoning. EQ is the most promising protective agent tested for bitterweed poisoning in sheep.

Animals↗

[Reduction of platelet serotonin in major depression (endogenous depression)].

Serum serotonin (5-HT) levels were measured in several patients with psychiatric disorders using high pressure liquid chromatography with electrochemical detection. A marked reduction in 5-HT levels was found in male and female patients with major depressive disorder, as compared to controls, but not in dysthymic disorder. These modifications may constitute biochemical changes suggestive of major depressive disorders; they could not be attributed to chronic antidepressant treatment.

Adult↗

Protective effects of antioxidants on bitterweed (Hymenoxys odorata DC) toxicity in sheep.

Antidotal effects of the 2 antioxidants butylated hydroxyanisole (BHA) and ethoxyquin (EQ) were evaluated in bitterweed (Hymenoxys odorata DC) toxicosis in sheep. Bitteerweed contains a toxic sesquiterpene lactone, hymenoxon, the toxicity of which is reduced by cysteine. Both BHA and EQ are known to induce hepatic glutathione production in rodents. Treatment of sheep with EQ (2.5 g/sheep/day for 9 days before poisoning) gave significant protection from toxic doses of bitterweed, but the protective effect of BHA was insignificant. Of 6 sheep given EQ in the feed, 5 survived 7 doses of bitterweed (4 g/kg/day or higher for 7 days), whereas 5 of 7 controls and 4 of 7 sheep given feed with added BHA died. The added EQ in the feed decreased the serum alkaline phosphatase activity and total protein, albumin, and calcium concentrations. Seemingly, EQ is the first protective agent with field application potential for bitterweed toxicity.

Animals↗

Toxicity and mutagenicity of hymenoxon, sequiterpene lactone.

The oral LD50 of hymenoxon in Swiss white mice was found to be 241 +/- 37 mg/kg. No significant sex differences were observed. Pretreatment of male mice for 3 days using doses of 50 and 100 mg/kg hymenoxon failed to alter significantly pentobarbital sleeping time. Hymenoxon was found to be a direct-acting mutagen in the Salmonella/mammalian microsome test. Urine samples obtained from hymenoxon-treated mice were found to be negative activity when tested directly and when incubated with beta-glucuronidase. Hymenoxon did not produce lethal DNA damage as measured in the Escherichia coli polA or Bacillus subtilis recombinational assays.

Animals↗

Antitumor agents. 44. Bis(helenalinyl) esters and related derivatives as novel potent antileukemic agents.

Bis(helenalinyl), bis(plenolinyl), bis(2,3-dihydrohelenalinyl), and bis(2,3,11,13-tetrahydrohelenalinyl) esters have been synthesized in an effort to elucidate the role of the two enone alkylating centers, beta-unsubstituted cyclopentenone and alpha-methylene gamma-lactone, as well as the significance of the diester linkage with respect to the enhanced in vivo P-388 lymphocytic leukemia antileukemic activity of bis(helenalinyl) malonate (2) against P-388 lymphocytic leukemia in the mouse. The bisesters (2-5; 7, 8; 10, 11) are, in general, more potent and less toxic than their corresponding parent alcohols (1, 6; 9; 14). The beta-unsubstituted cyclopentenone ring and the alpha-methylene gamma-lactone moiety in the bisesters play important roles for the enhancement of the P-388 antileukemic activity. Removal of the enone double bonds in both alkylating centers of 2 gave rise to inactive compounds. Except for 2, the potent antileukemic activity of the bis(helenalinyl) esters (3-5) appears to be independent of the ester chain length.

Animals↗

Influence of maternal dietary protein and fat levels on fetal growth in mice.

The influence of maternal dietary fat and protein levels during gestation on fetal growth was determined in LAF1/J mice. Dams were fed diets containing various levels of casein (ranging from 6 to 20%) and fat (either 5 or 15%). On the 18th day of gestation, all animals were sacrificed. Products of conception were examined, and maternal liver and fetal body compositional analysis was performed. It was found that low protein diets did not depress caloric intake, indicating that observed effects are due to differences in respective nutrient intakes rather than altered caloric intakes. Both the dietary protein and fat levels significantly influenced the fetal cellular growth (cell number rather than cell size) of LAF1/J mice. In addition, dietary fat influenced protein adequacy for satisfactory fetal cellular growth of these mice, probably by altering protein utilization. Optimal fetal cellular growth was obtained with the diet containing 20% casein and 15% fat as indicated by increased fetal protein, DNA, and RNA contents. Normal maternal liver lipid content in animals fed the 20% protein and 15% fat diet also indicates that this diet is optimal for gestational performance of LAF1/J mice.

Animals↗