The alpha-glyceryl ethers (AGE) are widely distributed in nature, occurring in many marine organisms, land animals, plants, and humans. A series of preliminary experiments was begun to determine whether these compounds possessed possible pharmacological actions against inflammation. Anti-inflammatory tests utilized both the granuloma pouch and cotton pellet granulation methods. Possible glucocorticoid activity was checked by measuring liver glycogen deposition. The effects of AGE on growth and weight of the animals was also noted. Neither selachyl dihemisuccinate sodium (selachyl DHSS) nor selachyl alcohol decreased the exudate formation in the granuloma pouch experiments, although excellent results were obtained with hydrocortisone. The high dose of the AGE was the equivalent of ten milligrams; low doses were three hundred and six hundred micrograms. Routes of administration used were oral, intraperitoneal, and subcutaneous. In the cotton pellet granuloma test, some indication of anti-inflammatory activity was obtained, in accordance with previous results in this and other laboratories. Selachyl DHSS decreased granulation tissue formation by only 11.7% in growing rats, which is not significant; however, in mature rats, there was a decrease of 29.5%. The dosage used in this series was 30 milligrams per kilogram daily by the subcutaneous route, A definite and comparatively large increase in liver glycogen deposition was observed in rats allowed to eat freely, when given selachyl DHSS subcutaneously. These glycogen values were decreased radically by a paired feeding study in which the treated rats were allowed to eat only the same amount of food as their paired control animal. Thus food intake appears to be one of the factors responsible for the increased glycogen storage. The AGE appear to affect growth only by the Intraperitoneal route of administration. In this case, the growth of rats was retarded by 29.6%. It was also noted that these animals ate somewhat less food than the controls, although this did not appear to be an important enough difference to account for the lack of weight gain. Throughout these experiments one of the most notable observations was a lack of uniformity of the results. Factors which appeared to cause a variability in the outcome of tests included the age of the animals, length of time of preoperative housing in new quarters, route of administration, and dosage. A non-linear relationship between dose and effect appears probable. / Medicine, Faculty of / Anesthesiology, Pharmacology and Therapeutics, Department of / Graduate

Caffeine has been shown to induce a transient hypertensive state in Wistar rats. The height to which the blood pressure rises in this caffeine-induced hypertension, and the duration of this hypertensive state was found to be dependent on the concentration of caffeine administered. Caffeine exposed to negatively ionized air was shown to undergo a loss in its pressor activity. This loss in pressor activity was found to be greater when the caffeine was exposed in solution than when it was exposed in the crystalline state. Once the blood pressures of Wistar rats were elevated with injections of caffeine and had again returned to normal levels there were no further rises in blood pressures with the administration of an equal number of injections of this drug. / Science, Faculty of / Botany, Department of / Zoology, Department of / Graduate

Extracellular action potentials and isometric contractile tension have been recorded simultaneously in vitro from uterine longitudinal smooth muscle of the pregnant cat, pregnant rabbit, estrogen-treated rabbit, and estrogen-treated rat. Action potentials were recorded from the surface of the muscle strips with glass electrodes having a large tip diameter. Tension was recorded with an RCA transducer. Spontaneous contractions are associated with a series of action potentials. During relaxation no electrical activity is observed. Electrical and mechanical activities were first recorded in Kreb's Ringer medium and then in sodium-poor media (replacement of sodium chloride with choline chloride or sucrose). Sufficient reduction in the external sodium concentration resulted in increased amplitude (peak to peak) of the biphasic action potential spikes. The duration of the peak to peak deflection and the maximum rate of potential, change remained unchanged. However, decrease in the external sodium concentration reduced the frequency of the action potentials, considerably in the cat, and less so in the rabbit and rat. The external sodium concentration was reduced in stepwise ; fashion to ½, ¼ , ⅛ its initial value. Each successive decrease in the external sodium concentration was accompanied by a prompt initial contraction, followed by very slow relaxation and subsequent resumption of spontaneous contractions accompanied by action potentials. With cat uteri reduction of the sodium concentration of the medium to a level of 15-20 mEq/1 resulted in a greatly prolonged contraction with eventual relaxation when tissues failed to contract. This paralysis was associated with cessation of action potentials. The electrical responses of uteri of the other two species (rabbit and rat) during exposure to sodium-poor media were similar to those observed with the cat uterus. However, the mechanical activity of rat and rabbit uteri in sodium poor media was different from that of the cat uterus. Decrease in the external sodium concentration below 25-30 mEq/1 usually resulted in prolonged contractions, and finally to complete failure of the tissue to relax (even after 2-2½ hours). Outbursts of action potentials at irregular intervals were seen in the initial stages of this persistent contraction but eventually action potentials also disappeared. It was difficult to reconcile these facts with the "Sodium Hypothesis". A selective inward flow of sodium ions probably cannot account for the initiation of action potentials in uterine smooth muscle since considerable reduction of the external sodium concentration (down to 15-20 mEq/1 in cat and 25-30 mEq/1 in the other two species) did not affect the characteristics of the action potentials in the expected manner. However, further reduction in sodium did result in electrical and mechanical inactivity. The view that an outward flow of intracellular anions might be responsible for depolarization (14) receives further support from the present studies. In addition to many differences from other types of excitable tissue (nerve, cardiac and skeletal muscle), uterine smooth muscle also shows considerable intra- and inter-species variation. / Medicine, Faculty of / Anesthesiology, Pharmacology and Therapeutics, Department of / Graduate

The behaviour reactions and the mechanisms involved in the injury and death of goldfish (Carassius auratus) exposed to chill temperatures have been investigated. Upon direct transfer to colder water goldfish exhibit an initial shock reaction followed by a primary chill-coma reaction with the latter reaction sometimes being followed, after apparently normal recovery, by a secondary chill-coma that ends in death. The first two reactions are believed to result from the effect of an excessive thermal stimulation of the cells of the central nervous system whereas the death that follows the secondary chill-coma is attributed to a disruption of the osmotic regulatory system. Within the range of size compared (three to ten centimeters and one to twelve grams), the tolerance to chilling temperatures of goldfish of the same or nearly the same age increases as the size of the fish increases. Statistical analysis of this relation shows a significant positive correlation between the survival times and the weight of the fish over their length as well as a significant negative correlation between the survival times and the surface area of fish over their weight. Several factors are thought to be involved in this effect of size of fish on tolerance to chill temperatures, viz., the insulation and surface area of fish relative to body mass in relation to the rate of heat loss and therefore to the body temperature; the gill surface area of fish relative to body mass in relation to the rate of abnormal osmotic passage of water subsequent to a disruption of the osmotic regulatory system. / Science, Faculty of / Zoology, Department of / Graduate

An attempt has been made to follow some of the metabolic changes occuring during acclimation of goldfish (Carassius auratus) to temperature. The goldfish were acclimated to certain temperatures and then reacclimated to different temperatures. The change in the unsaturation of their body fats was found using Wij's iodine value determinations. The fats of goldfish fully acclimated to different temperatures show different iodine values. In general the fats are more saturated at higher temperatures. Changes in the amount of unsaturation during the acclimation process have been followed. It has been found upon increase in temperature there is a decrease in the amount of unsaturation. / Science, Faculty of / Zoology, Department of / Graduate

A method is described for measuring the preferential responses of fish to alterations in the chemical and physical nature of their environment. Changes in the responses of young chum salmon at various stages of growth to alterations in the environmental sea water content have been studied. The effects of small alterations in temperature (2 to 3 C° ) and in rates of flow of environmental liquids on the fishes' responses to salinity change, were also studied. Preliminary experiments on the responses of coho salmon fry to sea water are described. The results indicate that changes in the environmental sea water content, such as those encountered in the estuaries of salmon streams may exert a directive influence on the seaward movement of chum salmon fry. / Science, Faculty of / Zoology, Department of / Graduate

An attempt has been made to alter the degree of unsaturation of the body fats of goldfish (Carassius auratus) and correlate these changes with any modifications of heat tolerance subsequently exhibited by the fish. The goldfish were fed three different diets each containing a fat of different degree of unsaturation. The fats used were pilchard oil (iodine value of 181.7), herring oil (iodine value of 128.4.) and lard (iodine value of 66.2). Heat resistance was tested by holding the fish at a constant high temperature and observing the time to death. Variations in the ability of the groups to withstand high temperature were then compared to differences in the degree of unsaturation of their extracted fats. It was found that while diet could effect changes in the degree of unsaturation of the goldfish fats to approximately 54% of the theoretical level, and that these changes in turn modified the heat resistance of the goldfish, no quantitative relationship was established. / Science, Faculty of / Zoology, Department of / Graduate

The mycotoxin zearalenone is known for its harmful effects on livestock reproduction. Animal exposure occurs through feed sources colonized by Fusarium species which produce the mycotoxin. Since regular screening procedures for zearalenone are not conducted on Western Canadian barley, a survey was carried out to test for possible significant levels of contamination. All samples were found to be negative at a detection level of 500 ppb; therefore, feeds formulated from the barley samples sources would not likely cause zearalenone toxicosis problems in livestock. Also, an ELISA method, Agri-Screen™, developed by Neogen Corporation (Lansing, Michigan) was tested and found to be a simple and economical method for pre-screening of feed samples in the field. To study direct toxicological effects of zearalenone on in vitro murine blastocyst development, murine embryos were cultured in medium (Ham's F-10 + estrous cow serum) containing various levels of the mycotoxin. The critical concentration range for zearalenone to cause detrimental effects on blastocyst development was determined to be between 70-160 μg/ml medium. Additionally, a concentration effect on the length of time required to exert deleterious actions was demonstrated. At mycotoxin concentrations of 500 μg/ml medium and above, blastocysts degenerated after 6 h of culture. At a lower concentration level of 160 μg/ml, blastocysts were not affected until 28 h of culture. In order to investigate the direct toxicological effects of zearalenone on in vitro porcine pre-implantation embryo development, attempts were made to develop a successful culture system. Since a suitable system was not developed, toxicological studies were not possible. Possibly, steps in the recovery process could have resulted in detrimental effects before the embryos were placed in culture. Alternatively, the media chosen (Ham's F-10 + estrous cow serum; Minimum essential medium + fetal calf serum) may not be suitable for in vitro culture of porcine pre-implantation embryos. Finally, at a zearalenone concentration level (250 μg/ml medium) found to cause degeneration of murine blastocysts, the in vitro maturation of bovine oocytes in Tissue Culture Medium 199 was not affected. It was suggested that the surrounding cumulus layer acts as a barrier to prevent the mycotoxin from directly acting on the oocyte. / Land and Food Systems, Faculty of / Graduate

In Part I, a modified synthesis of radio-labelled secodine (68) and its incorporation into vindoline (7) is described. In a model study, for the synthesis of side-chain labelled 3-ethylpyridine (74), [2-² H]-(3'-pyridyl)-ethane was achieved from the correspondingly labelled 3-acetylpyridine by desulphurization of the intermediate thioketal (93). In a second study, [1- ³H]-(3'-pyridyl)- ethane was synthesized by treating 3-acetylpyridine with sodium borohydride-³H. The resulting alcohol (95) was acetylated, and hydrogenolysis achieved the desired product. The ester alcohol (74) was coupled to [1- ³H]-(3'-pyridyl)-ethane and the resulting pyridinum salt (90) was reduced to the corresponding piperdeine ester (80) in a "one-pot" synthesis. The conversion of (80) to [19-³H]-secodine was achieved by a known procedure. In two experiments, [19-³H, ¹⁴C0₂CH₃]-secodine (68)(³H/¹³C ratios = 3.00 and 1.54) was administered to Catharanthus roseus plants. The vindoline (7) which was isolated was shown to have been biosynthesized from the entire secodine molecule (³H/¹³C = 3.31 and 1.35 respectively). In Part II, a degradation scheme designed to achieve the isolation of the N-methyl group of uleine (1) is described as well as preliminary results from an investigation into the biosynthesis of uleine (1) and olivacine (4). Variously radio-labelled forms of tryptophan (15), anthranilic acid and secodine (18) were administered to Aspidosperma pyricollum root segments and whole plants. The uleine (1) which was isolated was found to be inactive in all experiments. Variously radio-labelled forms of tryptophan (15), anthranilic acid and secodine (18) as well as ¹⁴CH₃-methionine (30) was administered to Aspidosperma australe plants. Uleine (1) and olivacine (4) was isolated. The only incorporation that could be demonstrated was that of ¹⁴CH₃ methionine (30) into uleine (1) to the extent of 0.168% and 0.147%. The isolation of the N-methyl group from (1) showed that it contained 97% and 98% of the activity. In Part III, the attempted synthesis of compounds of the preakuammicine- and stemmadenine-series is described. A new method for the C-18 deoxygenation of curan derivatives using Birch reduction conditions was achieved. Also, a modification of the Oppenauer oxidation of the curenol (36) to achieve improved yields of the aldehyde (37) and nor-fluorocurarine (39) was developed. The introduction of a carbomethoxy group into the C-16 position of the curan aldehyde derivatives (44) and (50) using a base and methylchloroformate was unsuccessful. Also, the introduction of cyanide into position C-16 of the indole alcohol (52) or indole acetate (57) via the corresponding chloroindolenines was unsuccessful. The synthesis of product (60), which is believed to be identical with preakuammicine aldehyde (7), was achieved. This material could not be converted into akuammicine (5) or stemmadenine (4). Only the dehydrated indolenine (72) could be obtained. The ring-opening reaction of the corresponding thioacetal derivative (73) yielded the decarboxylated indole thioacetals (75) and (76). / Science, Faculty of / Chemistry, Department of / Graduate

The neuroanatomical consequences of chronic ethanol ingestion were investigated using neuropsychological tests selectively sensitive to damage in circumscribed brain areas-~~~ The performance of a group of male alcoholics (N=15) with a self-reporte?- drinking history less than ten years, and a group of male alcoholics (N=15) with a self-reported drinking history of ten or more years was compared to a group of male nqnalcoholic controls (N=15). l.A--multiple covariate analysis was done prior to analyzing group differences to determine what influence three covariates (age, education and socioeconomic status) had on the group performances. Results from this analysis indicated that some of the tests were significantly influenced by age or education but no significant Group x Covariate interactions were observed. /JCn agreement with numerous neuropathological findings, the late stage alcoholics were found to be principally deteriorated on the tests of frontal lobe functioning, followed by lesser impairment on the tests of temporal lobe functioning, with no evidence of impairment on the tests of parietal lobe functioning~ No significant group differences were found betv1een the early stage alcoholics and the controls~ Small but negative correlations were found between both the duration of alcoholism and the average amount of alcohol consumed with each of the neuropsychological measures given in this study.