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Biomedical subjects

R F Keeler

Publications and source records attributed to R F Keeler.

At least 37 records · Page 2Linked to original sources

Individual animal susceptibility and its relationship to induced adaptation or tolerance in sheep to Galega officinalis L.

Oral administration of Galega officinalis L to sheep demonstrated a marked variation in individual animal susceptibility to the toxic effects of the plant. As little as 5 g/kg of dried ground plant induced moderate tracheal frothing in 1 ewe while nearly 5 times that amount failed to elicit any recognizable toxic effects such as frothing, pulmonary edema or hydrothorax in others. Ten g/kg induced severe effects in 3 ewes. Ewes administered levels of plant between 5 and 24 g/kg had toxic effects whose severity was often unrelated to level administered. There was no apparent difference in average severity of clinical signs of toxicity nor pathologic lesions to challenge doses of 24 g/kg of the plant between groups of ewes with an immediate previous history of increasing doses of the plant and others with no history of ingesting the plant. Previously reported apparent induced adaptation or tolerance to G officinalis L in some animals is more likely to have been a result of the extreme variation in individual animals susceptibility.

Adaptation, Physiological↗

Maternal and fetal toxicity of poison hemlock (Conium maculatum) in sheep.

Poison hemlock (Conium maculatum) was toxic to pregnant ewes and their fetuses when fed during gestation days 30 through 60. Maternal effects included trembling, muscular weakness in the neck initially, then progressing to the limbs, ataxia, frequent urination and defecation, and death. Convulsive seizures were not observed. Fetotoxic effects included excessive flexure of the carpal joints with lateral deviation in the front limbs and kinked tails. At term, 7 of 11 lambs had varying degrees of the limb abnormalities, but all lambs appeared clinically normal at 8 weeks after parturition.

Animals↗

The nature of congenital limb defects induced in lambs by maternal ingestion of Veratrum californicum.

Groups of ewes were gavaged Veratrum californicum root-rhizome material over various 3 day periods during early gestation and their late-term fetuses examined. Resulting from gavage near day 29 were shortening of fetal metacarpal, metatarsal and tibial bones. Fibulae bowed severely when tibiae were shortened. Medial bowing of forelimbs at the fetlocks resulted from unequal shortening of fused metacarpi. Severe joint articular surface defects and arthrogryposis were present in some of the lambs from treatment ewes. There was bilateral and twinning symmetry in limb bone shortening. Severity of limb defects varied with gestational stage at time of gavage.

Animals↗

Toxicity of Thermopsis montana in cattle.

Cattle had severe signs of toxicosis when gavaged dried ground Thermopsis montana (false lupine, poison bean, mountain thermopsis) at doses of 0.6-2.8 g/kg/day in a water suspension. Signs included depression, anorexia, swollen eye lids, arched back, tucked abdomen, rough hair coat, and in extremis a prolonged recumbency lasting up to 9 days. Plant potency varied among collections. Total alkaloid doses in collections eliciting severe signs varied from 1.1-11.3 mg/kg/day. There were 5 major alkaloids in each collection that varied in concentration among the collections perhaps accounting for variation in severity of signs elicited. Four of the alkaloids were identified by GC retention time, MS fragmentation patterns and OR analysis as N-methylcytisine, cytisine, (-)-thermopsine, and (-)-anagyrine. Measurements showed a very marked increase of 10X-20X in levels of certain serum enzymes--SGOT, CPK, and LDH that persisted during the period of maximum clinical signs.

Alkaloids↗

Toxicosis from and possible adaptation to Galega officinalis in sheep and the relationship to Verbesina encelioides toxicosis.

Galega officinalis L (goatsrue), a plant introduced from Europe and found in abundance in northern Utah, was tested for toxicity in ewes (Ovis aries). Clinical signs of poisoning induced by doses as small as 0.8 g of dried plant/kg body weight/day included dyspnea, anoxia, and foaming nasal discharge. Pathologic signs in animals that died following overdose of the plant included severe hydrothorax, generalized lung congestion, foamy exudate in bronchioles and trachea, epicardial and endocardial petechiation, and pericardial effusion in severely affected ewes. Microscopic lesions included a severe diffuse alveolar and interlobular edema. No significant alterations in blood constituents were observed in treated animals. Ewes administered the plant on several consecutive days developed an apparent adaptation to the toxin of the plant and were thereby able subsequently to tolerate levels 5-10 times the pre-treatment lethal dose with no discernible adverse clinical or pathological effects.

Animals↗

Congenital tracheal stenosis in lambs induced by maternal ingestion of Veratrum californicum.

Seven of nine lambs born to six ewes gavaged with Veratum californicum root and rhizome material on days 31, 32 and 33 of gestation died from asphyxia within 5 minutes after birth. Five of the seven were autopsied and found to have severe stenosis of the trachea. None of the 12 lambs born to seven control ewes had tracheal stenosis. The defect was characterized by lateral flattening of the trachea throughout its entire length. Cartilaginous tracheal rings were reduced in number, nonuniform in size and shape, irregularly spaced, and with abnormal orientation. The rings were thinner than those of control tracheas, and had zones of chondrogenesis on outer and inner surfaces which were of equal width, rather than of unequal width as in controls. The smaller size and relatively noncurved shape of the cartilaginous rings of the stenotic trachea resulted in a nondistended lumen.

Animals↗

Inhibition of limb chondrogenesis by a Veratrum alkaloid: temporal specificity in vivo and in vitro.

It has been demonstrated that jervine, a steroidal alkaloid derived from plants of the genus Veratrum, exerts teratogenic effects in several animal species. Defects were restricted to structures which depend upon normal chondrogenesis for their development. Here we report studies of the temporal specificity of cellular sensitivity using limb bud mesenchyme cells obtained from Day 10 mouse embryos. These cells, when grown as high-density "spot" cultures, undergo chondrogenesis in vitro. Prior to differentiation, exposure of limb cell cultures to jervine suppressed subsequent accumulation of cartilage proteoglycans. Treatment after differentiation had no significant effect. Additionally, there was a genetic component to jervine sensitivity: C57BL/6J mice were sensitive, whereas NIH Swiss-Webster mice were insensitive. This strain-dependent difference was observed both in vivo and in vitro, supporting the validity of limb mesenchyme spot cultures as a model for jervine-induced teratogenicity. Our studies indicate that jervine acts specifically during an early phase of the differentiation of mesenchyme into cartilage. It is likely that a specific stem cell population is the target tissue of this compound.

Animals↗

Induction of cleft palate in newborn pigs by maternal ingestion of poison hemlock (Conium maculatum).

Cleft palates were induced in newborn pigs of gilts fed Conium maculatum seed or plant during gestation days 30 through 45. Twelve of 23 newborn pigs born to 3 gilts given Utah-grown C maculatum seed and 9 of 12 newborn pigs born to a single gilt given the fresh Utah spring-growth C maculatum plant had cleft palates. The cleft palates ranged from a unilateral cleft, involving only 1 side of the palate, to a full bilateral cleft. Brachygnathia was also observed in some of these newborn pigs with cleft palate. Other malformations were not observed. Chemical analysis of seed and plant samples indicated that gamma-coniceine was the responsible teratogenic alkaloid. A daily dose of plant or seed that provided greater than or equal to 1.07 mg of gamma-coniceine/kg of body weight fed to gilts during the 30th through the 45th day of pregnancy resulted in teratogenic effects.

Alkaloids↗

Congenital skeletal malformations induced by maternal ingestion of Conium maculatum (poison hemlock) in newborn pigs.

Skeletal malformations were induced in newborn pigs from gilts fed Conium maculatum seed or plant during gestation days 43 through 53 and 51 through 61. The teratogenic effects in groups dosed during gestation days 43 through 53 were more severe than those in groups dosed during the later period, with many newborn pigs showing arthrogryposis and twisted and malaligned bones in the limbs and with 1 pig showing scoliosis and deformity of the thoracic cage. The pigs born to gilts given C maculatum during gestation days 51 through 61 had excessive flexure primarily in the carpal joints, without scoliosis or bone malalignment in the limbs. The teratogenicity of poison hemlock depends on the alkaloid concentration and content. Based on the data presented, we speculate that gamma-coniceine is the teratogenic alkaloid in the poison hemlock fed to the gilts.

Abnormalities, Drug-Induced↗

Isolation of teratogenic alkaloids by reversed-phase high-performance liquid chromatography.

Reversed-phase high-performance liquid chromatography was used for both analytical and preparative separations of several steroidal alkaloids which occur in extracts of Veratrum californicum. The inclusion of 0.1% trifluoroacetic acid in the mobile phase improved the efficiency of the chromatography and the solubility of the compounds in aqueous acetonitrile. Nuclear magnetic resonance was used to assist the identification of the isolated steroidal alkaloids. The effect of the interaction of trifluoroacetic acid with the alkaloids could be clearly seen by changes in the chemical shifts in the nuclear magnetic resonance spectra.

Chromatography, High Pressure Liquid↗

Teratogenicity in swine of the tobacco alkaloid anabasine isolated from Nicotiana glauca.

Nicotiana glauca, wild tree tobacco, induces arthrogrypotic congenital defects in piglets similar to those induced by Nicotiana tabacum, common tobacco. The present work was conducted to isolate the principal alkaloid of N. glauca, anabasine, in large quantity and good purity and to test the teratogenicity of the compound in pigs. The isolated compound was established to be anabasine and to be of suitable purity by chemical characterization. It proved to be teratogenic. Typical arthrogrypotic defects were induced in 21 of 26 offspring (three of three litters) when dams ingested 2.6 mg of the compound per kg body weight twice daily during the 43rd-53rd days of gestation. Of three dams dosed with 1.66 g/kg/day of the dried plant material during the 43rd-53rd days, one delivered deformed offspring representing one-third of all offspring in that group. These arthrogrypotic defects induced by anabasine were indistinguishable clinically from defects induced by either N. glauca or N. tabacum. In addition, anabasine at a dose of 2.6 mg/kg twice daily or N. glauca plant material at 1.66 gm/kg daily induced cleft palate in over three-fourths of offspring (100% of litters) when dams ingested either during the 30th-37th days of gestation or during longer periods that included those days.

Abnormalities, Drug-Induced↗

Structure and stereochemistry of steroidal amine teratogens.

Studies of teratogenic steroidal alkaloids from Veratrum and Solanum have shown that those bearing a basic nitrogen atom in ring F, shared or unshared with ring E, with bonding capabilities alpha to the steroid plane may be suspect as teratogens. Examples of steroidal alkaloids which produce terata but, until recently, have been of uncertain structure include muldamine and the isomeric 3, N-diformylsolasodines. The correlation of their structures with the structure-terata relationship developed by Keeler and Brown is discussed. A brief introduction to teratogenicity is presented.

Molecular Conformation↗

Teratogens in plants.

Many compounds synthesized by plants are known to be teratogenic in laboratory animals, but only a few have been shown by feeding trials to produce terata in livestock. Studies of plant teratogens affecting livestock have not moved forward in a systematic nor rapid way because of the logistical problems connected with such experiments in large animals. Information that has accumulated can be conveniently separated into three categories: (1) known teratogens in known teratogenic plants, (2) known teratogenic plants with unidentified teratogens, and (3) suspected teratogenic plants. Included in the first group are the teratogens from Lupinus, Veratrum, Conium and Leucaena genera; in the second group are included the Astragalus , Nicotiana and Trachymene genera; and in the third group are included Datura, Prunus , Sorghum and Senecio genera. Total available information in each case varies, but in a few instances considerable fundamental as well as practical information is now known. Research has provided enough information in a few instances to enable elimination of the practical problem or significant reduction in incidence.

Abnormalities, Drug-Induced↗

Teratogenicity and toxicity of wild tree tobacco, Nicotiana glauca in sheep.

Collections of Nicotiana glauca containing from 0.45-1.14 mg anabasine per gram dry weight of plant induced moderate - severe toxic signs in pregnant ewes and teratogenic effects in their offspring when the ewes were given single daily doses of the plant that provided from 1.66 - 3.42 mg anabasine per kg body weight. Toxic signs included excess salivation, irregular gait, wobbling while walking or standing, recumbency and death. Teratogenic effects occurred in offspring from animals fed those doses the 30th - 60th day of gestation. Terata expression included limb defects such as a fixed excessive carpal flexure with or without lateral or medial rotation of fore or rear limbs, lordosis, irregular shaped head or cleft palate.

Abnormalities, Drug-Induced↗

Specific craniofacial defects induced by jervine in the cultured rat embryo.

The Veratrum-derived steroidal alkaloid, jervine, induces cyclopia and limb malformations in sheep, and various other craniofacial malformations in several other mammalian and avian species. In the present study, the question whether jervine acts directly or indirectly on mammalian embryos to produce malformations and the nature of the target tissue or cells were examined using whole-embryo cultures of the CD rat. Embryos were explanted into culture at the presomite, early neurula stage and cultured in the presence or absence of jervine for 48 hours. Jervine (at 1-5 micrograms/ml) induced an oblong-head appearance and ventrally displaced optic vesicles, with little or no other effects observed on overall growth and development. The specific target tissue in the embryo was found to be the cranial neuroepithelium. This specificity of action is quite unusual since most teratogens examined in whole-embryo culture to date have various nonspecific effects on embryonic growth and differentiation.

Animals↗

Congenital deformities in swine induced by wild tree tobacco, Nicotiana glauca.

Eighty-four of 147 piglets from 20 purebred Hampshire gilts that were bred to purebred Hampshire boars and then fed Nicotiana glauca during various periods of gestation were congenitally deformed. The deformities occurred in piglets from gilts on experiments in 3 separate years during which 4 different collections of the plant were fed. The deformities included various arthrogrypotic limb deviations and palate closure defects. Arthrogrypotic limbs had excessive flexure of pastern or carpal joints, medial rotation, or a combination of these effects. There were no gross visceral defects nor histopathologic changes in examined tissue. Expression of deformities varied as a function of gestation period. Palate closure defects, for example, occurred only in offspring from sows fed prior to gestation day 35. The limb deformities induced by N. glauca in piglets were similar to those reported by others and assumed, or known from feeding trials, to have been induced by maternal Nicotiana tabacum stalk ingestion. By contrast palate defects were not reported in piglets from maternal N. tabacum ingestion, but were of high incidence in the present work from maternal N. glauca ingestion. The suspect teratogen, anabasine, the principle piperidine alkaloid of N. glauca, varied in concentration among the collections fed. However dosages were maintained in each animal regardless of collection fed that resulted in daily toxicity signs indicative of anabasine toxicosis. The signs included depression, tremors, third eyelid closure, irregular gait and recumbency.

Abnormalities, Drug-Induced↗

Teratogenic effects of Nicotiana glauca and concentration of anabasine, the suspect teratogen in plant parts.

Nicotiana glauca consumption by pregnant cows or sows produced arthrogryposis and other congenital deformities in their offspring. The plant material administered contained 0.11% anabasine, the suspect teratogen. But concentration varied among various plant parts; woody material was very low but leaves and bark were high. The anabasine was identified by mass spectral fragmentation analysis.

Anabasine↗