Intrahepatic microvascular changes in carbon tetrachloride-induced cirrhotic livers in the rat. Liver cirrhosis was produced in the rat by combined carbon tetrachloride-phenobarbitone treatment, and the microcirculation in the cirrhotic liver was observed by a quantitative in vivo transillumination technique. The total sinusoidal flow in the observed region of the cirrhotic liver did not differ significantly from that in the normal liver, despite the reduced number of sinusoids and the increased portal venous pressure. The cirrhotic liver also presented a fast-velocity population of portal and hepatic venules and sinusoids in addition to the normal slow-velocity population. The possible mechanism of these "arterialized" microvessels is discussed.


Effect of the ligation of hepatic artery on the microcirculation in the cirrhotic liver in the rat. The significance of the hepatic arterial supply in the intrahepatic microcirculation in normal and carbon tetrachloride-induced cirrhotic livers was studied by dye injection method and by ligation of the hepatic artery. The in vivo distribution of dye injected into the hepatic artery evidenced the presence of arterio-venous shunts in the cirrhotic liver. When the hepatic artery of the cirrhotic liver was ligated, the elevated portal venous pressure dropped significantly, and the fast-flowing population of microvessels and sinusoids in the bimodal frequency distribution plot disappeared. The fast-flowing microvessel and sinusoids appeared to be the "arterial" microvessels and sinusoids, and they were converted into the slow-flowing venous channels after hepatic arterial ligation. The transmission of arterial pressure via the A-V shunts may be of greater significance in the pathophysiology of portal hypertension than previously believed.


Serological evidence of inter-epidemic infection of feral pigs in New South Wales with Murray Valley encephalitis virus. The sera of 617 feral pigs, collected from three widely separated areas of northern and central New South Wales, were examined for antibody to Murray Valley encephalitis (MVE) virus and to Ross River virus. Haemagglutination-inhibition (HI) antibody was detected to MVE in 58% of sera and to Ross River virus in 15% of sera. Neutralization tests suggested that the MVE HI antibody resulted from infection with MVE virus in the summers of 1971-1972 and 1972-1973 when the virus was not known to be active in New South Wales. These same tests suggested that more than one flavivirus infected the feral pigs in the summer of 1973-1974 and that Kunjin virus was active in the summer of 1975-1976.


Basophil leucocytes in cutaneous hypersensitivity reactions in nematode-infected guinea-pigs. Cutaneous hypersensitivity reactions were elicited in guinea-pigs infected with the parasitic nematode Trichostrongylus colubriformis by injecting them with a crude extract of T. colubriformis fourth-stage larvae. The reaction was characterized by early oedema and pronounced cellular infiltration, initially with neutrophils but later with mononuclear cells, basophils and variable numbers of eosinophils. Because basophils have been implicated as effector cells in the protective immune response of guinea-pigs to this nematode, the capacity to elicit a basophil-rich cellular infiltrate in infected animals might be a useful assay for T. colubriformis protective antigen(s).


The relationship between the transfer of immunoglobulins, sodium and potassium into mammary secretion of the parturient ewe. The experiment comprised two sections. First, radiotracer techniques were used to study the metabolism of IgG1 and IgG2 in 5 non-pregnant and 4 pregnant ewes. In the pregnant ewes, the rates of synthesis for IgG1 and IgG2 were similar to the rates observed in non-pregnant animals. However, the irreversible loss of IgG1 was significantly greater than IgG2 in pregnant ewes and IgG1 in non-pregnant ewes. Additionally, it was found that most of the IgG1 and virtually all of the IgG2 in mammary secretion was serum derived. Secondly, the levels of sodium, potassium and lactose and the selective index for IgG1 in mammary secretions of 5 pregnant ewes were monitored over the parturient period. The values of all the measures remained relatively constant until one day before parturition. From one day pre-partum, the levels of potassium and lactose in mammary secretion began to increase and had risen 2-3 fold by 5 days post-partum. Over the same period, the selective index for IgG1 decreased 20 fold, wehreas the level of sodium fell from approximately 32 mmol/l to 18 mmol/l. The concentration of IgG1 in plasma slowly declined from approximately 23 g/l to 15 g/l over the last 10 days of pregnancy. During the parturient period, the decline in plasma IgG1 levels, in comparison with IgG2, without alteration in the rates of synthesis of either immunoglobulin, supports the hypothesis that selective transport of IgG1 into mammary secretion occurs without degradation. The results also indicate that the transport of sodium and potassium into mammary secretions are altered over the parturient period.


Inhibition of intestinal uptake of amino acids by unconjugated bile salt. The unconjugated bile salt, sodium deoxycholate, at a concentration of 0.5 mM was shown to inhibit the intestinal uptake of the amino acids L-glycine, L-leucine, L-proline, L-lysine and L-tyrosine in rats in vitro. This effect was acutely reversible except for the basis amino acid L-lysine and is therefore not simply due to tissue damage. These results, and the recent finding that sodium deoxycholate inhibits intestinal absorption of amino acids in vivo, suggest that impaired intestinal amino acid transport may contribute to hypoproteinaemia in patients with bacterial overgrowth in the upper small intestine in whom deoxycholate is present in the small intestinal lumen in excessive concentrations.


Fibrinogen-like proteins and fibrinolytic proteins in a saline eluate from sucrose washed bovine erythrocytes. Plasminogen and a spectrum of fibrinogen-like proteins are found in a saline eluate of sucrose washed bovine erythrocytes. It seems probable that these proteins are membrane bound in vivo, and, as such, their physiological significance is discussed.


The effects of 2-phenylalanine 8-lysine vasopressin (octapressin) on blood vessels in the rat tail. Isolated perfused segments of the ventral rat tail artery have been used in this study to investigate the effects of octapressin on vascular smooth muscle and its interaction with other vasoconstrictor agents. It was found that repeated octapressin administration was associated with tachyphylaxis and that the onset and rate of development of this phenomenon could be modified by cocaine. The constrictor action of catecholamines, electrical stimulation and serotonin on the rat tail artery was potentiated by octapressin. Mechanisms which may be involved in octapressin tachyphylaxis are discussed.


Infection rates and outbreaks of disease due to Babesia argentina in unvaccinated cattle on 5 beef properties in south-eastern Queensland. Serums from unvaccinated groups in 5 herds of beef-cattle in South-East Queensland were tested for antibodies to Babesia argentina at intervals while the cattle were increasing in age from about 6 months. An indirect fluorescent antibody test was used. Infection rates, indicating the proportions of the groups that had been exposed to tick-transmitted infection were 49.2, 56.9 and 69.1% for cattle aged approximately 6, 12 and 18 months, respectively. The degree to which cattle were infested with the vector, Boophilus microplus, was estimated. There appeared to be a strong correlation between infection rate and tick incidence. Four serologically negative animals died of either confirmed or suspected babesiosis during the sampling period. Sickness was observed in 5 others. The serological status of 57 changes from negative to positive without symptoms being observed, indicating relatively low mortality and morbidity rates in the enzootic situation studied.


The induction of parturition using dexamethasone. Premature parturition was successfully induced with a preparation containing 2 dexamethasone esters in 709 out of 716 cows and heifers of gestational periods ranging from 3 to 8 months. A mortality rate of 38.5% was experienced among calves produced by induced cows, while only a 2.7% mortality rate occurred among 4,330 calves from normal parturitions. Approximately 50% of all treated cattle retained foetal membranes, but with apparently no ill-effect. The treatment apparently did not affect milk production, post-calving fertility, and the overall incidence of dystocias in cows and post-calving diseases. Three deaths occurred among induced cattle, but they could not be attributed to the treatment. It was concluded that the technique may prove to be of great value in dairy cattle, in seasonal milking areas. The induction of premature parturition was attempted in 736 cows and heifers.  In each animal, an initial im injection of 32 mg dexamethasone was followed by no more than 3 injections of 16 mg at approximately 10-day intervals.  Most animals were 5-8 months pregnant at time of treatment.  Premature parturition was successfully induced in 709 (99%);  64.5% required only the initial injection, 33.9% required 2 injections, and 1.6% required 3.  14.2% of the fetuses were stillborn, and 24.3% of the calves died before 4 weeks of age, compared with a 2.7% mortality rate among 4330 calves from normal parturitions.  Approximately 50% of the treated cattle retained fetal membranes with no apparent ill-effect.  The treatment did not effect milk production, postcalving fertility, and the overall incidence of dystocias in cows and postcalving diseases.  This technique, particularly in the seasonal milking areas, may be a valuable management procedure.


Vaccination with Babesia argentina in 5 beef herds in south-eastern Queensland. Observations on the use of a vaccine containing Babesia argentina in 5 partly susceptible beef herds in south-eastern Queensland were made on 1,029 female breeding cattle over a period of 4 years. Groups averaging about 20 heifers were given 0, 1, 2 or 3 vaccinations. Incidence derived from groups experiencing clinical attacks were 17.9% for unvaccinated cattle and 1.2% for vaccinates. Increasing the number of vaccinations did not appear to increase protection. The one clinical manifestation of infection with B. bigemina was associated with a concurrent reaction to vaccination with Anaplasma centrale. There were no cases of haemolytic anaemia in new-born calves. No severe reactions followed primary vaccination, but 2 revaccinated animals became sick.


Tuberculosis-like granulomas in cattle caused by Actinomycetes. From 280 lesions from normal cattle at slaughter, 10 were found to contain thin, branching, Gram-positive organisms belonging to the Nocardia/Streptomyces group and this was confirmed by culture in 3 cases. Macroscopically the older lesions were similar in appearance to those of tuberculosis. Microscopically, all lesions contained numerous foreign body giant cells and thin, branching organisms. The older lesions also contained areas of necrosis, caseation, mineralisation and fibrosis.


A pathological and bacteriological study of the mammary glands of beef cows in north Queensland. Pathological and bacteriological studies were made of a total of 507 udders from slaughtered beef cows originating from various locations in northern Queensland. Incidence of supernumerary tests was 28%, their distribution being caudal (80%), intercalary (8%), ramal (5%) and anterior 1%; 6% had both caudal and intercalary teats. They were seen most frequency in British breed and British-Brahman crossbreds (each 29%) and less frequently in Brahmans (22%). Gross lesions were found in 11.4% of cows; these lesions comprised cysts (2.8%), leech infestation (1.0%), mastitis (0.6%), teat occlusionsion (3.9%), papillomatosis (1.2%), cutaneous horns (0.4%), teat fistulas (0.4%), teat trauma (0.6%) and udder fibrosis (0.6%). Mastitis was obviously low in occurrence, while leech infestation and teat occlusion were relatively frequent in cows from one area. Bacteriological examination of single quarters in 364 cows, revealed infection in 154 (42.3%). A total of 199 isolations were made of which 3.5% were recognised mastitis pathogens. Pathological conditions were found, and bacterial isolations were made more frequently in aged than in mature or young cows.


Polyarthritis of pigs in western Australia: the role of Erysipelothrix rhusiopathiae. A 3-month survey of polyarthritis in pigs was carried out at 3 metropolitan abattoirs in Western Australia. The incidence of total carcass condemnations for polyarthritis was 0.46% and partial condemnation for "arthritis" 1.66% of 15,919 pigs. It was demonstrated that the majority of joint lesions found in either total or partial condemnations were identical pathological conditions, usually differing only in the number of joints involved. Erysipelothrix rhusiopathiae was isolated from the joints of 63% of pigs condemned for polyarthritis; it appears that this organism is the most significant aetiological agent of polyarthritis in Western Australia. The role of mycoplasmas has not been determined. The groww and microscopic lesions seen in affected joints were those of a non-suppurative proliferative polysynovitis. The stifle and elbow joints showed the highest incidence of the most severe lesions. Pigs condemned for polyarthritis over a 6-month period were traced back to 125 farms. Eighty per cent of pigs that had been condemned for polyarthritis had not been vaccinated against erysipelas or were of doubtful vaccination status. Only 3% of pigs had been vaccinated as recommended. The results would suggest that erysipelas vaccine is not directly implicated in the pathogenesis of polyarthritis. The majority of pigs condemned for polyarthritis originated from poorly managed small piggeries run as sideline enterprises. There was no significant pattern to the geographical distribution of polyarthritis cases in Western Australia. Likewise, no significant realtionship was established between polyarthritis condemnations and either breed, sex or bodyweight of thepigs involved.


Complications following a ruptured bladder in a 60-day-old foal. The death of a 60-day-old foal due to massive haemorrhage associated with erosion of the left umbilical artery into the left ureter is reported. Surgical repair attempts are described together with post-mortem findings.


The "Dag" and swollen abaxial midpiece spermatozoan defects in bulls.


Some thoughts on academic institutions.

Some thoughts on academic institutions.


The veterinary profession and the future in Australia.


Supply and demand for veterinarians in Australia.


Osteoarthrosis of the elbow joint in dogs.


Babesiosis of a horse in Australia.

Babesiosis of a horse in Australia.


The species of flies reared from struck sheep in south-eastern Australia.


Mimosine in Leucaena leucocephala is metabolised to a goitrogen in ruminants.


Avulsion of the tibial tuberosity in the greyhound.


Congenital bovine epizootic arthrogryposis and hydranencephaly in Australia. Distribution of antibodies to Akabane virus in Australian Cattle after the 1974 epizootic. At the end of the 1974 epizootic of bovine congenital arthrogryposis and hydranencephaly in south-eastern New South Wales, an Australia-wide serological survey (about 4,000 serums) was made to determine the ditribution of cattle possessing serum neutralising antibodies against Akabane virus. Eighty per cent of the serums from cattle in northern Australia (Western Australia, Northern Territory, and Queensland) were positive. A detailed study in the epizootic area in New South Wales (particularly around Bega) showed that 80 to 100% of serums from cows in herds in this area possessed neutralising antibodies. The animals possessing antibodies extended as far south as Genoa in north-eastern Victoria, and as far west as Darlington Point on the Murrumbidgee River. There were no positive herds along the Murray River, where an outbreak of the mosquito-borne disease Murray Valley encephalitis occurred in 1974. Serums tested from cows in the rest of Victoria, South Australia, south-western Western Australia, and Tasmania were negative. Arthrogrypotic calves born in Tasmania and south-western Western Australia were not associated with the presence of Akabane virus. In Papua New Guinea, serums collected from cattle at Boroka, Lae, and Goroka did not possess neutralising antibodies. The distribution of cattle possessing antibodies in Australia would fit a spread of the virus by Culicoides brevitarsis, a biting midge from which Akabane virus had been isolated on three occasions. The possibility of other vectors, as well as C. brevitarsis, was suggested by the presence of cows possessing antibodies at Alice Springs, where this biting midge has not been found. Possibly most cattle in northern Australia become infected early in life. The epizootics in New South Wales could occur when seasonal conditions allow a southerly extension of virus-infected C. brevitarsis which feed on susceptible pregnant animals. C. brevitarsis also bites sheep, and both neutralising antibodies to Akabane virus and congenitally deformed lambs have been observed in the epizootic area. An understanding of the distribtuion of Akabane virus and C. brevitarsis, a possible Australian vector for bluetongue virus, may prove useful if bluetongue should enter Australia.


A review of respiratory agents associated with disease of sheep, cattle and pigs in Australia and overseas.


Custom collection and processing of bull semen. Three systems of custom collection and processing of semen are reported. In the first system, semen was collected, evaluated and partially diluted and returned to a central Government laboratory for final dilution and freezing. In the second system the procedure was similar to the first except the semen was returned to the author's laboratory for final dilution and freezing. System 2 reduced the failure to process rate from 20.4% to 6%, but did not significantly improve fertility (64% v's 68% for systems 1 and 2 respectively). A single-stage diluent was used in system 3 and the time from collection to freezing in the author's laboratory was reduced. Both the failure to process rate (0.1%) and the fertility rate of the semen (76%) improved over the other 2 systems.


Control of reproductive function of cattle using cloprostenol. Two intramuscular injections of cloprostenol were given 12 days apart to synchronise oestrus in a group of 47 heifers and following the second injection, 38 were found in oestrus in 2 to 6 days, 1 in 11 days and 8 in 22 to 27 days. The conception rate of the synchronised animals was 24 of 38 (63%). Twenty-nine heifers were given injections of cloprostenol in late pregnancy and parturition was induced with a latent period of 6 +/- 3 days when injected at 266 to 267 days. When injected at 254 to 262 days, 3 of 16 heifers (19%) were unresponsive and the remainder calved with a latent period of 12 +/- 6 days. The survival rate of calves from induced heifers was 71% with 80% survival from controls. Approximately 50% of the induced heifers, and none of the controls, retained their foetal membranes.


The incidence of dystocia in Dorset Horn ewes. The incidence of dystocia was recorded in flocks of Dorset Horn ewes lambing in winter, spring or summer and grazed at 8, 12 or 16 ewes per hectare over a ten-year period. The mean incidence of dystocia was 34% of ewes lambing. Significantly more ewes experience a difficult birth at winter (35%) and spring (39%) than summer (26%) lambings. Neither stocking rates nor pre-lambing ewe bodyweight were related to the incidence of dystocia. The incidence of dystocia was significantly greater in 2-year-old ewes than in any other age group. Foetal size was of major importance in the occurrence of dystocia. Approximately 60% of single and twin born lambs that had birth weights greater than 4.5 kg had a difficult birth whereas in lambs weighing less than 3.2 kg only about 10% were affected. Thirty-six per cent of all perinatal mortality was attributable to dystocia. Approximately 27% of single and 73% of twin presentations in assisted births were abnormal, 31% of all lambs assisted had 1 or both legs retained, 4% had posterior (including breech) presentations and the remaining 8% were of varied forms of malpresentation. Significantly more single male than female lambs experienced a difficult birth and the mean birth weights of single, twin and triplet born lambs that had a difficult birth were greater than the birth weights of singles, twins and triplets born normally.


Simulated natural infection of chickens with Australian lentogenic newcastle disease virus and subsequent challenge with virulent virus. A total of 291 eight-week-old chickens were exposed to chickens infected with either of two Australian lentogenic strains (V4 and AVL NDV-1) of Newcastle disease virus (NDV). At 3 weeks after exposure, all chickens exposed to V4 infected chickens had developed haemagglutination-inhibition (HI) antibody. All chickens exposed to AVL NDV-1 virus infected chickens had developed HI antibody 5 weeks later. This sudden late appearance of HI antibody, to titres higher than those observed with V4 chickens, was explained by V4 virus being introduced to the AVL NDV-1 group of chickens. When groups of these chickens were challenged with Roakin virus (mesogenic NDV) at 3 weeks and Fontana 1083 virus (viscerotropic velogenic NDV) and Texas GB virus (neutrotropic NDV) at 3, 5, 10 and 21 weeks only three chickens developed clinical illness one of which died. These chickens were one AVL NDV-1 chicken contact challenged with Fontana 1083 virus at 3 weeks, one V4 chicken oronasally challenged with Texas GB virus at 5 weeks and one V4 chicken challenged oronasally with Fontana 1083 virus at 10 weeks. Susceptible non-vaccinated chickens died soon after challenge. Challenge by oronasal infection with 10(7.0) ELD50 of virus or contact with susceptible infected chickens enabled virulent virus to be isolated from most chickens and was accompanied by a large anamnestic increase in serum HI antibody.


Some factors affecting the measurement of blood inorganic phosphorus in cattle. Sixty cows were studied over 3 days, to determine the effect of site of sampling, handling stress and the tranquilliser xylazine hydrochloride, on blood inorganic phosphorus (P). Jugular venous and coccygeal mixed arterial and venous blood were compared. Saliva was also analysed for P. Buffered and unbuffered trichloracetic acid (TCA) were compared as deproteinising agents. Tail blood P was consistently 12% (range 7-17%) higher than that from the jugular vein (P less than 0.01). This difference decreased when jugular blood was sampled first but increased following administration of the tranquilliser. The drug significantly (P less than 0.01) increased the concentration of P in the saliva on 2 out of 3 days. The possible role of salivary P in affecting jugular P values, is discussed. The buffered TCA analyses gave consistently lower P readings than the unbuffered analyses. The magnitude of this effect was time dependent, being greater when the samples were left for 4 days (P less than 0.05) than when analysed the next day. There was a very strong correlation between plasma and whole blood P. It was concluded that the tissue of choice in phosphorus studies is tail blood (plasma or serum) treated with buffered TCA.


Scrotal mesothelioma in a bull.

Scrotal mesothelioma in a bull.


An attenuated vaccine against bovine ephemeral fever.


Suspected aspergillosis in a dog.

Suspected aspergillosis in a dog.


Treatment of pregnancy toxaemia in ewes by induction of parturition.


Lesions in sheep and guinea pigs pen fed parasitised annual ryegrass (Lolium rigidum)


Artificial insemination of horses.

Artificial insemination of horses.


Stimulation of the development of infective Babesia bovis (= B. argentina) in unfed Boophilus microplus larvae.


Equine hydatidosis in Australia.

Equine hydatidosis in Australia.


Anthelmintic treatment of young beef cattle in the Wallum region of south-eastern Queensland. An anthelmintic treatment trial on 125 Brahman-British crossbred 2 to 4 month old calves was undertaken on Wallum country in south eastern Queesland. The calves were divided into 5 experimental groups, grazed together and treated as follows for 17 months: Group 1--Untreated controls; Group 2--Monthly levamisole--niclosamide on 4 occasions; Group 3--Monthly levamisole until 1 month after weaning; Group 4--Levamisole 3 to 6 weeks after saturating rains; Group 5--No levamisole--niclosamide as for Group 2. The mean body weight gains for cattle in Groups 1 to 5 were 95, 124, 105, 121 and 97 kg respectively. Four cattle were lost from each of Groups 1 and 3 and five from Group 5. Most of these losses occurred towards the end of the second summer rainfall season. Faecal egg count maxima were recorded around weaning in untreated groups and during the summer rainfall period of both the first and second summer in all groups. The most prevalent nematode species encountered were H. placei, Cooperia spp and O. ostertagi. It is recommended that in this region cattle under 2 years of age should receive anthelmintic treatment at least in autumn and spring.


A study of nephritis of beef cattle in North Queensland. The incidence of subclinical nephritis in beef cattle slaughtered at a Townsville abattoir during the period 1970-73 was 3.8 percent. A rising incidence coincided with the wet summer of 1973-74 reaching a peak of 8.2 percent thereafter in June 1974. All but 1 of 100 cases macroscopically identified as nephritis and studied histologically proved to be of interstitial type in which lymphocytic infiltration predominated. Follicular lymphoid hyperplasia was also a common feature. The exceptional case showed proliferative glomerulonephritis. Leptospires were isolated from 16 percent of cases cultured, and observed in 8 percent of specimens examined histologically. All of the 8 isolates subjected to serogrouping were L. pomona. Of the animals tested serologically against 2 serotypes 66.2 percent were positive to L. pomona (33.8 percent), L. hardjo (18.9 percent) or both (13.5 percent). The total incidence is significantly higher than in other random surveys carried out in the region. These findings suggest that Leptospira sp. are a major cause of bovine nephritis. Since other pathogens endemic in the area, for example, Babesia and Theileria sp., may be capable of causing nephritis in cattle an accurate assessment of the aetiology of that form of renal disease in north Queensland cannot yet be made.


Humpy back of sheep. Clinical and pathological observations. Humpy back, a disease of Merino sheep in western Queesland, occurs during mustering for shearing. It is usually seen in summer 6-10 weeks after substantial rainfall and is thought to be caused by the ingestion of a toxic plant. The disease is characterised clinically by a short-stepping, stilted gait of the hind limbs, followed by lowering of the head, arching of the back and inability to continue walking. Histopathological examination of 8 cases from 5 properties revealed a Wallerian degeneration of the white matter throughout the length of the spinal cord with the ventral and lateral columns most severely affected. A similar degenerative change was seen in the posterior cerebellar peduncles of 3 of the sheep. A more severe hind limb incoordination with more extensive degeneration of the white matter of the spinal cord, medulla and cerebellum was seen in a case of humpy back of two years duration. Similar, but much milder, spinal cord lesions were found in apparently unaffected sheep from the same group as the sheep affected with humpy back on 2 properties. Severe myodegeneration of hind limb muscle groups was seen in 3 affected sheep. It was thought to be associated with the long rail journey (1500 km) to the laboratory after the sheep were affected in the field.


Studies on the prevalence of ringworm among horses in racing and breeding stables. A survey of 568 horses in training and 2,535 horses on breeding farms has shown 32 percent of the horses in training to be clinically affected while only 1.1 percent of breeding horses were affected with pathogenic dermatophytes. The majority of lesions on racing horses were located on the girth areas. Trichophyton equinum var. autotrophicum, M. canis and M. equinum were found to be restricted to racing horses only. M. gypseum occurred in racing, riding and breeding horses.


A standardised haemagglutination inhibition test for porcine parvovirus antibody. Basic variables of the haemagglutination inhibition (HI) test for porcine parvovirus antibody were investigated. Nonspecific serum inhibitors were satisfactorily removed without loss of specific antibody when undiluted serum was adsorbed with 25 percent kaolin in borate saline at pH 9.0. Natural haemagglutinins in test serums could be completely removed using 0.1 ml of packed erythrocytes to 0.6 ml of kaolin treated serums. Adsorption of prediluted serum resulted in a depression of specific antibody titres. Highest HI titres were obtained using guinea pig erythrocytes, following incubation of virus-serum mixtures for 18 hours at 4 degrees C, 3 hours at 25 degrees C or 2 hours at 37 degrees C. Micro- and macro-tests gave comparable HI titres.


Leptospiral agglutinins in dogs in Sydney. From June 1971 to June 1972, sera from 600 dogs in Sydney were tested for leptospiral agglutinins by a rapid slide agglutination method. The end-point titre was taken at 50 percent agglutination of the live organisms. Forty-one samples (6.8 percent) had a significant leptospiral titres (100 or greater) and 5 of these reacted to 2 serotypes. Thirty serums (5 percent) contained agglutinins against L. copenhageni, and 6 (1 percent) against L. pomona, while a few samples reacted against hardjo, tarassovi, australis, grippotyphosa or pyrogenes serotypes. No significant titres were found to L. canicola, L. hebdomadis, L. autumnalis or L. bataviae.


Suspected equine infectious anaemia in a foal.


Solanum esuriale, a possible cause of humpy back in sheep.


Extension of the range of Rhipicephalus sanguineus in Western Australia.


The role of dietary fat in alleviating fatty liver and kidney syndrome in broilers.


Reactivation of recent or remote memory before electroconvulsive therapy does not produce retrograde amnesia.


Dissociation of odor and taste in shock-avoidance learning.


Parallel but independent effects of pentobarbital and scopolamine on hippocampus-related behavior.


A comparison of free-operant shuttle avoidance in Mongolian gerbils and albino rats.


Differentiation of response biases elicited by scopolamine and d-amphetamine: effects on habituation.


An experimental test of diaschisis.

An experimental test of diaschisis.


Social and genotypic aspects of open-field behavior of two lines of mice.


The response of male guppies (Poecilia reticutata, Peters) to repeated exposure to an open field.


Adaptation and aversive brain stimulation. II. Effects of current level and pulse frequency.


Conditioned taste aversion: oral and postingestional factors.


Adaptation and aversive brain stimulation. III. Excitability characteristics of behaviorally relevant neural substrates.


The CNS weighting of external and internal factors in feeding behavior.


The role of individual recognition by odors in the social interactions of the Mongolian gerbil (Meriones unguiculatus).


The influence of calcium and beta-sympathomimetics on force-frequency relationship and resting potentiation in guinea pig papillary muscle. The influence of various calcium concentrations and of the beta-sympathomimeticum orciprenaline (Alupent) on the force-frequency relationship and on the resting potentiation of isolated isometrically contracting papillary muscles of the guinea pig heart was studied. Elevation of Ca concentration and/or addition of orciprenaline augment the amplitudes of contractions elicited by rhythmical stimulation, but they diminish the amplitudes of test contractions elicited after a test interval of 1-2 s. It is suggested that calcium and orciprenaline strengthen the calcium loss from the sarcoplasmic reticulum out of the cell during a prolonged interval.


Maximal rate of pressure rise and time parameters in the right ventricle under isovolumic conditions. Investigations in the canine heart-lung preparation. In contrast to the left ventricle, the maximal rate of intraventricular pressure rise in the right ventricle does not occur within the isovolumic phase of the contraction. Occlusion of the pulmonary artery by inflation of a balloon during the diastole causes an isovolumic systole of the following heart beat. The canine heart-lung preparation was used to test whether dP/dtmax measured isovolumically and the peak ventricular pressure Pmax as well as the parameters derived are useful indices for the contractile state of the right ventricle. Changes only of the aortic pressure influence neither dP/dtmax, Pmax and the time values from the onset of contraction to dP/dtmax (t-dP/dtmax), nor the time to Pmax (t-Pmax). A rise in heart frequency leads to an increase in dP/dtmax and Pmax at lowered enddiastolic pressure. It does not influence the time interval t-dP/dtmax and t-Pmax. With augmented diastolic filling, dP/dtmax as well as Pmax increase, and t-dP/dtmax and t-Pmax are extended. The present study suggests that dP/dtmax and Pmax measured isovolumically provide accurate and practical measurements of right ventricular contractility, provided that changes in enddiastolic pressure and heart frequency are taken into account. The time parameters are found to be not useful.


A classification of tumor development based on an analysis of enzymes in tissue sections of hypernephroid carcinoma in man. Sections of hypernephroid carcinoma from 20 cases were investigated for aldolase isozymes A and B by a mixed aggregation immuno-cytochemical technique, and for the brush border membrane enzymes aminopeptidase and alkaline phosphatase by conventional histochemical techniques. It was found that the cases could be grouped into four types: type 1 (1 case) contained all 4 enzymes; type 2 (7 cases) contained all enzymes except aldolase-B; type 3 (7 cases) possessed aldolase-A and one brush border membrane enzyme; type 4 (5 cases) contained only aldolase-A. The aldolase-A concentration in all tumor cells was higher than that in proximal tubule cells, whereas the concentration of the two brush border enzymes was lower. In cases tydolase-B and/or higher amounts of the brush border enzymes than the surrounding cells. No correlation was observed between clear cell and granular cell hypernephroid carcinomas or the invasiveness or the nuclear polymorphism of the tumors on the one hand with their enzyme type on the other. These histological enzyme analyses suggest that most, if not all, hypernephroid carcinomas are derived from kidney proximal tubule cells and that the tumor cells then progressively lose aldolase-B, and subsequently the brush border enzymes, but at the same time producing more aldolase-A. The presence of the enzyme-rich patches suggest different patterns of proliferation and differentiation among the tumor cell population. Three tumors other than hypernephroid carcinoma were also examined in this way. The results suggest that histoenzymological analyses are of general applicability in studies of tumor progression. They should also be useful for biopsy and aspiration cytology.


Quantitative electron microscopic study of capillaries in diabetes mellitus. A systematic microscopic examination of all elements of the capillary wall was performed on quadriceps muscle biopsies from 9 diabetic patients and 8 controls. The capillary basement membrane (CBM) was markedly thicker in diabetics; it consisted of several lamellae and contained large vacuoles which were never observed in non-diabetic subjects. Large magnifications revealed fibrils in greater number and markedly larger in diameter in diabetics, these accounting for a considerable volume of the CBM and the adventitia and increased diameter and thickness of the capillary wall, without encroaching on the lumen. The intracellular fibrils in pericytes and endothelial cells were also larger and thicker in diabetic subjects. The prevalence of fibrillar material in the vascular disease of diabetes mellitus suggests the importance of research into possible measures to arrest fibril formation.


The maximal rate of pressure rise in the right ventricle of isolated canine hearts in isovolumic and auxotonic systole under various hemodynamic and inotropic conditions. The influence of changes in pre- and afterload, heart frequency and inotropic state on the difference between right ventricular dP/dtmax measured isovolumically and auxotonically has been investigated in canine heart-lung preparations. Isovolumic systoles were obtained by occluding the pulmonary artery during diastole by balloon inflation. The mean difference between isovolumic and auxotonic dP/dtmax is 40 mm Hg/sec (15 to 80 mm Hg/sec) - i.e. about 10% of the value measured auxotonically. Neither hemodynamic (pre- and afterload changes) nor inotropic changes influence this difference. Heart rate increase from 140 to 200/min also has no effect. This corresponds with the unchanged time interval between pulmonary valve opening and right ventricular dP/dtmax under all these conditions. Only heart rates below 100/min seem to enhance the difference between isovolumic and auxotonic dP/dtmax. Since the difference is not altered at various inotropic states, the result of this study leads to the conclusion that the relative changes in contractility could be slightly overestimated by using the auxotonic dP/dtmax for inotropic assessment. Practically, this inexactitude is within the error range of the method. Therefore, the use of dP/dtmax measured auxotonically does not introduce a marked disadvantage related to isovolumic dP/dtmax for assessment of relative inotropic changes of the right ventricle in isolated hearts.


The significance of DNA flow-through fluorescence cytophotometry for the diagnosis of prostate carcinoma. Flow-through fluorescence cytophotometric determination of nuclear DNA content was employed for the diagnosis of prostate carcinoma. Fine needle aspiration biopsy material from the prostate of 220 patients was used for study. A false negative rate of 11.4% and a false positive rate of 29.7% were obtained when the results of flow-through photometry were compared with those of traditional cytodiagnosis. It was found that 4.5% of the specimens were unsuitable for cytologic diagnosis and 10.9% for flow-through cytophotometry. False negative DNA histograms may be due to two factors: either the number of tumor cells is small or there are tumor cells whose nuclear DNA content does not differ from that of a normal cell population. False positive findings result from proliferating cells in inflammatory activation. Errors in preparation of the material and mechanical mistakes, such as cellular clumping and coincidences, are less likely causes. The greater percentage of specimens which were inadequate for cytophotometry was due to the large number of cells needed for a utilizable flow-through photometric histogram. The high rate of false negative and false positive results (11.4% and 29.7%, respectively) argues against using flow-through photometric nuclear DNA determination for the diagnosis of prostate carcinoma.


Evaluation of physical performance by rectangular-triangular bicycle ergometry and computer-assisted ergospirometry. A great deal of information about physical adjustment to work can be obtained from quantitative stress testing. Maximal stress limited by symptoms of exertional intolerance is the concept of the 2 min duration work increment test (rectangular-triangular exercise test). Compared to steady-state work tests strict observation of the standardized procedure- and computer assisted evaluation of ergospirometric parameters offer innovatory opportunities: (1) the test is of short duration (8-14 min), (2) the subjects recover rapidly, even from an exhausting test, (3) one is more likely to be able to observe plateauing of VO2, should determination of maximal VO2 be desired, (4) adaptation to increasing work rates and maximal work capacity is assessable, (5) computer technics provide on-line assessment of aerobic and anaerobic power in quantitative terms, (6) measurements proved to be highly reproducible, (7) the relationship between variables such as increments of heart rate and systolic blood pressure, respiratory minute volume, oxygen uptake during the early phase of the non-steady-state condition and the index of anaerobic power, and the influence of factors such as work load and work output, has been studied to derive standard values. Soft-ware programs have been designed to estimate deviation of parameters actually measured from standard values in terms of multiples of the standard deviation of the standard regression line. In particular, evaluating oxygen uptake during short time-intervals (0.5 min) provides information about adequate adaptational forces of the cardio-circulatory system. Energy that is not accounted for by reactions involving the VO2 measured is computed by substracting the caloric equivalent of oxygen uptake during work exceeding the steady-state level during rest from the energy demand to sustain a given work load aerobically. This index of anaerobic power is defined in kcal, cal/kg body wt., and as a percentage of the total amount of energy required (moderately trained athletes 350-500 cal/kg; sedentary people 200-300 cal/kg). A close relationship to parameters of metabolic acidosis (base excess) exists. It is concluded that the physical performance of sedentary people, athletes and patients with impaired cardio-pulmonary function can be more precisely qualified in quantitative terms by means of computer assisted rectangular-triangular ergospirometry. Results obtained in patients with diseased conditions must be carefully interpreted, their condition suggesting the use of more invasive investigations to reveal the pathophysiologic mechanism.


Hemodynamic response to digitalization in patients with hypertensive cardiovascular disease. Twenty-eight patients with hypertensive cardiovascular disease (HCD) and incipient myocardial dysfunction underwent hemodynamic studies at rest and during exercise before and 30 minutes after administration of 0.6 mg beta-methyl-digoxin intravenously. Measurements were made during right heart catheterization with a balloon-tipped catheter. The hemodynamic changes after administration of digitalis did not demonstrate a consistent and uniform improvement of cardiac performance in all patients with HCD and myocardial dysfunction. When separating 11 patients with pervious myocardial infarctions or documented coronary artery disease (CAD) (= Group I) from the remaining 17 subjects without clinical and/or angiographic signs of CAD (= Group II), there were significant differences in the hemodynamic response to digitalis: In Group I, pulmonary artery wedge pressure (PAWP) after digitalis decreased only slightly and insignificantly from 8.7 to 7.4 mm Hg at rest and from 27.6 to 26.4 mm Hg during steady state exercise. Cardiac output (CO) remained essentially unchanged with a tendency to decrease after digitalis: 5.9 vs. 5.8 L/min at rest and 11.5 vs 11.1 L/min during exercise. At rest, even patients of Group II showed only minor decrease of PAWP from 8.8 to 7.2 mm Hg; during exercise these patients demonstrated marked improvement of cardiac performance with a significant decrease of PAWP after digitalis from 27.8 to 22.3 mm Hg (p less than 0.01). With one exception, there was a more or less pronounced reduction of PAWP after the drug was given. No significant change of CO after digitalis was measured in this group: 6.2 vs. 5.9 L/min at rest and 13.4 vs. 13.5 L/min during exercise. The different hemodynamic patterns of responders and non-responders to the glycoside will be discussed.


The influence of dihydroergotamine on adenosine-induced and reactive coronary vasodilation. Interaction of dihydroergotamine and coronary vasodilation. The influence of dihydroergotamine on adenosine-induced and reactive vasodilation after long and short periods of coronary artery occlusion was investigated in thoracotomized dogs. Adenosine-induced vasodilation (intracoronary administration) and vasodilation after long periods of coronary artery occlusion (25-35 beats) were similarly influenced, i.e. decreased by the i.v. administration of 10 mug/kg dihydroergotamine. By contrast vasodilation after short periods of coronary artery occlusion (4-7 beats) tended to be increased. This difference in response is thought to arise from two distinct mechanisms of coronary vasodilation after coronary artery occlusion depending on the duration of the occlusion period. The vasodilation after short periods of coronary artery occlusion possibly corresponds to physiological autoregulation. With longer periods of coronary artery occlusion an additional, consecutive mechanism is called into action.


Experimental hypothyroidism: depression of myocardial contractile function and hemodynamics and their reversibility by substitution with thyroid hormones. Hemodynamics and myocardial muscle mechanics have been studied in 22 euthyroid and 60 hypothyroid cats in which experimental hypothyroidism has been produced by thyroidectomy 61 days prior to the examination. Left ventricular to body weight ratio was altered due to a decrease in left ventricular weight and an increase in body weight. Heart rate, cardiac output and cardiac index were decreased (by 12-15 per cent), whereas stroke volume remained unchanges. Peak systolic pressure of the left ventricle was moderately decreased, the other pressures were in the normal range. There was a marked and significant reduction of isovolumic contractility indices indicating a depression of myocardial contractility in situ by 20-27 per cent. The isolated ventricular myocardium exhibited decreases of isotonic muscle shortening, of maximum isometric tension development and of the rates of both, isotonic shortening and isometric tension development by 12-35 per cent. Force-velocity relationships of contraction and relaxation were depressed to lower values of contraction and relaxation velocity as well as of maximum isometric muscle tension. The alterations in myocardial muscle mechanics and hemodynamics were completely reversible following substitution of the hypothyroid group with physiological doses of L-thyroxine (5 mug/kg/day for 8-18 days). Excess increases of parameters of myocardial performance were found following substitution of the hypothyroid group with L-thyroxine (500 mug/kg/day) in accordance with the induction of experimental hyperthyroidism in these animals. The results demonstrate impaired myocardial contractility and hemodynamics in experimental hypothyroidism. These changes are completely reversible by substitution with L-thyroxine in accordance with a reversible thyroid cardiomyopathy. The cellular mechanisms responsible for the altered cardiac activity in experimental hypothyroidism are discussed.


Effects of fendilin hydrochloride (sensit) on systemic and coronary hemodynamics and on retrograde coronary blood flow in dogs. Fendilin hydrochloride was tested in 9 normal anesthetized dogs and in 7 anesthetized dogs with acute coronary artery occlusion. The compound (1 mg/kg i.v.) caused a slight short lasting increase in coronary blood flow together with an increase in cardiac metabolic rate for oxygen of similar magnitude. There was no measurable effect on collateral conductance. No noteworthy effects on systemic hemodynamics were observed.


Studies on the regulation of myocardial blood flow in man. I.: Training effects on blood flow and metabolism of the healthy heart at rest and during standardized heavy exercise. In a comparative study 11 athletes and 11 untrained students were investigated at rest, of these 6 trained and 5 untrained individuals during exercise as well. Myocardial blood flow was measured by the argon method. Myocardial oxygen consumption, myocardial substrate uptake of glucose, lactate, and free fatty acids and cardiac output were determined by the direct Fick principle. Exercise was standardized according to 65% of an individual's maximal oxygen uptake (delta VO2 max). Coronary flow reserve was determined by dipyridamole injections. All measurements were made during hemodynamic and respiratory steady-state conditions with the subject in a supine position. At rest, myocardial blood flow and myocardial oxygen consumption were significantly lower in trained subjects compared to the untrained ones. These differences were more pronounced during heavy exercise. They cannot be explained completely by hemodynamic parameters. - During exercise, myocardial substrate uptake shifted to a predominant lactate uptake of almost 90% of total substrate uptake. Total substrate uptake as well as lactate uptake correlated significantly with myocardial oxygen. - Coronary flow reserve was lower in the trained group. It is concluded that the heart muscle of a trained individual requires less energy at a given work load than in the untrained state.


Studies on live attenuated mumps vaccine. II. Follow-up study on the efficacy of Biken Vaccine. Clinical and serological follow-ups were made on 18 children for 3 years and on another 18 children for 4 years, after immunization against mumps by attenuated mumps vaccine; Biken vaccine. To evaluate the protective efficacy of the vaccine, matched controls were studied during the same period. Serological examinations revealed that 77% of the vaccinees and 93% of the controls were infected with mumps after close contacts with mumps patients. Regular mumps was contracted by 88% of the controls, but none of the vaccinees. There was no substantial decrease in the antibody titers in unexposed vaccinees after vaccination.


Effect of hormone treatment on genital herpetic infection in mice.


Antitumor effects of interferon on transplanted tumors in congenitally athymic nude mice.


Isolation and purification of D-alanyl-meso-2, 6-diaminopimelic acid endopeptidase of Streptomyces L-3 enzyme using soluble substrates of known chemical structure from Lactobacillus plantarum cell wall digests. D-alanyl-meso-2, 6-diaminopimelic acid (D-Alanyl-meso-A2pm) endopeptidase was isolated and purified from a crude Streptomyces L-3 enzyme preparation by ion exchange chromatography and isoelectric focusing in a density gradient. During its purification, its hydrolytic activity was assayed on cell walls of Lactobacillus plantarum ATCC 8014 and soluble glycopeptides and peptides, of known chemical structures, prepared enzymatically from these cell walls. A fraction with an isoelectric point of pH 7.9 cleaved the bond between the carboxyl group of the D-alanine residue at the C-terminal in one peptide subunit and one of the two amino groups of the A2pm residue in the neighboring peptide subunit. Unlike the crude enzyme, the endopeptidase in this fraction showed no N-acetylmuramyl-L-alanine amidase, A2pm carboxyamide amidase or proteinase(s) activity and it was immunologically homogeneous.


Studies of the cell envelope of Vibrio para-haemolyticus A55: isolation and purification of bag-shaped peptidoglycan (murein sacculus). The peptidoglycan (PG) component of the envelope of Vibrio parahaemolyticus A55 was studied in relation to the salt dependency of the organism. A bag-shaped PG (murein sacculus) was isolated and purified by digestion of freshly harvested intact whole cells with sodium dodecyl sulfate (SDS) and then protease treatment. Whole cells lyze in salt-deficient or hypotonic conditions and this can be measured by turbidometry. Lysis is prevented by addition of 0.35 M NaCl or acidification to below pH 4.5. The isolated PG or murein sacculi did not seem to require salt for structural integrity. The PG obtained by the same method from subcellular fractions, that is, envelope fractions, prepared either with or without NaCl, was extensively fragmented. The main amino acids and amino sugars of the murein sacculus were glucosamine, muramic acid, alanine, glutamic acid, 2, 6-diamnopimelic acid (A2pm) in a molar ratio of 0.4:0.6:1.7:0.9:1.0. A polyglucose, which was only degraded by Pseudomonas isoamylase,and poly-beta-hydroxybutyrate (PHBA) were found in the murein sacculus. The murein sacculus of the organism is discussed in comparison with PG of other gram negative bacteria and marine or halophilic microorganisms.


Genetic control of heat sensitivity and activity level of murine arylsulfatase B. BALB/c male mice possess twofold higher kidney p-nitrocatechol-SO4 arylsulfatase B than do A/HeJ male mice; however, their liver arylsulfatase activities are comparable. Twentyfold-purified kidney arylsulfatases B from these two strains have similar Michaelis constants, electrophoretic mobilities, pH optima, and inhibitor profiles; however, the BALB/c enzyme is more heat stable than the A/HeJ enzyme. BALB/c, C3H/HeJ, DBA/2J, and SWR/J mice share an autosomal allele, As-1a, which apparently determines the heat-stable arylsulfatase B, while A/HeJ and C57BL/6J mice possess the As-1b allele, which determines the heat-sensitive enzyme. A second autosomal locus, Asr-1, determines liver arylsulfatase B activity. C57BL/6J mice carry the Asr-1a allele, which results in high liver activities, while C3H/HeJ mice are homozygous for the low-activity allele, Asr-1b. Male mice generally have 30-40% higher kidney activities than females; however, female kidney arylsulfatase activities rise and actually surpass those of males during late pregnancy and lactation.


Multiple structural genes for the alpha chain of canine (Canis familiaris) hemoglobin. There are two forms of the alpha chain of canine hemoglobin differing only at residue 130. One form (taualpha) contains threonine at this position, the other (Aalpha) contains alanine. Studies of two Labrador dog families strongly support the existance of multiple alpha-chain structural genes as the basis of the alpha-chain heterogeneity. There must be at least one gene locus for Talpha and one for Aalpha; the exact number of loci has not been determined. Two other dog breeds, the Basenji and the Beagle, also have both Talpha and Aalpha chains.


The enzymatic deficiency conditioned by the shrunken-1 mutations in maize. Evidence is presented to show that the Sh locus specifies sucrose synthetase in the developing endosperm of maize. The sh/sh/sh endosperm possesses less than 10% sucrose synthetase activity as compared to the normal Sh/sh/sh endosperm. The residual enzyme activity in five independently derived mutant genotypes is attributable to a protein molecule of different electrophoretic and immunochemical specificities that is presumably independent of the sh locus. Sucrose synthetase activity in the embryo in both the genotypes is electrophoretically indistinguishable from the one present in the mutant endosperm. Mutant endosperm has a reduced starch content as compared to the normal. This observation constitutes genetic evidence supporting a critical role for sucrose synthetase in starch biosynthesis.


Chromosome number and flavonoid synthesis in Briza L. (Gramineae). Following a recent reidentification of the apigenin C-glycosides in diploid Briza media, the luteolin C-glycosides were reexamined and three acylated derivatives of a luteolin C-glycoside were found. In an attempt to identify the chromosome or group of chromosomes responsible for the change in flavonoid synthesis from 4'-hydroxy- in diploid plants to 3',4'-dihydroxyflavone C-glycosides in autotetraploid plants, leaf flavonoids of artificially produced aneuploids of B. media were examined. Among these plants three different leaf flavonoid profiles were recognized: diploid, tetraploid, and a "modified" tetraploid pattern. All the aneuploids with the normal or "modified" tetraploid pattern were trisomic for one of the small acrocentric chromosomes. Induced polyploids of other Briza species were usually found to have similar flavonoid patterns in the two chromosome races. Flavonoid sulfates were found in three South American species but are absent from all the European species.


Analysis of repetitive DNA in three species of Gossypium. The rate of reassociation of denatured DNA was determined for two selected diploid species, Gossypium thurberi (D genome) and G. arboreum (A genome), and one allotetraploid species, G. hirsutum (AD genome). The relative genome size and DNA content of the chromosomes of the diploids were A greater than D. Renaturation curves indicated that the differences in genome sizes were due primarily to the repetitive DNA content.


Identification of wheat (Triticum aestivum L.) chromosomes with genes controlling the level of nitrate reductase, nitrite reductase, and acid proteinase using the Chinese Spring-Hope substitution lines. The levels of nitrate reductase, nitrite reductase, and acid proteinase were compared in the primary leaves of 8-day-old wheat seedlings of Chinese Spring, Hope, and the 21 disomic substitution lines of Hope in Chinese Spring. Two chromosomes, 7B and 7D, were considered to contain genes controlling the level of nitrate reductase. Substitution of Hope chromosome 7B caused a highly significant increase in the in vitro stability of nitrate reductase. Nitrite reductase appeared to be controlled by two major genes, located on chromosomes 4D and 7D, and two minor genes, located on chromosomes 3D and 5A. In the case of acid proteinase, substitution of chromosome 1D caused a significant reduction in enzyme activity.


Allelic expression in intergeneric fox hybrids (Alopex lagopus x Vulpes vulpes). I. Comparative electrophoretic studies on blood enzymes and proteins in arctic and silver foxes. Twenty-seven blood enzymes and proteins, whose structures are presumably controlled by at least 33 genes, were assayed in Arctic and silver foxes by starch gel electrophoresis. Two types of protein and enzyme electrophoretic patterns were distinguished: one exhibiting a single enzyme, the other several isozymes. The two fox species were found to differ in seven of the 27 enzymes and proteins studied: glucose-6-phosphate dehydrogenase, adenylate kinase, erythrocyte carboxylesterase, diaphorase, prealbumin, transferrin, and albumin. No differences were established between the species for the other enzymes and proteins. The data are interpreted as evidence for the existance of a set of enzymes and proteins differentiating the Arctic from the silver fox.


Correlation between structural variation and activity of murine kidney beta-galactosidase: implications for genetic control. Two closely linked regulatory genes have been reported to control activity levels of beta-galactosidases in murine tissues. The specific effects of these genes on murine glycolipid metabolism have not been elucidated. A/HeJ kidney 4-methylumbelliferyl-beta-galactosidase exhibited lower thermostability than the corresponding C57BL/6J and SWR/J enzymes. This altered response to heat segregated with the Bgsh allele among progeny derived from backcrosses of F1 (A/HeJ; SWR/J) mice to the respective parental strains. Restriction of the heat-sensitive A/HeJ beta-galactosidase to kidney tissue suggests that it is not determined by the Bgs locus, since the latter appears to be expressed in all tissues. More likely, the Bgs region of chromosome 9 contains a gene cluster consisting of a number of regulatory and structural loci. The proposed structural genes share affinity for the artificial substrates commonly employed for their assay but may differ in their relative affinities for glycosphingolipid substrates. Presence of the Bgsh allele results in an increase of kidney GM1-ganglioside-beta-galactosidase; however, galactosylceramide-beta-galactosidase appears unaffected by this allele.


Genetic control and distribution of leucine aminopeptidase in the cultivated chenopods (Chenopodium) and related weed taxa. Isozymes of leucine aminopeptidase (LAP) in leaf tissue of the cultivated chenopods (Chenopodium quinoa and C. nuttalliae) and their sympatric weedy relatives (C. hircinum and C. berlandieri) can be electrophoretically resolved into a sum total of five anodally migrating bands. These are the products of two unlinked gene loci which apparently reflect genetic duplication via allotetraploidy. Accessions from the Andean and Mexican areas of cultivation are characterized by a lack of electrophoretically detectable variation. Andean weed and cultigen accessions appear to be genetically identical at both Lap loci, as do weed and cultigen material from Mexico. The two cultigens, and their sympatric weeds, can be differentiated by variation at the Lap-B locus, whereas they are monomorphic at Lap-A. This locus is, however, highly polymorphic in weedy C. berlandieri populations of western North America.


On the detection of functional and structural enzyme mutants by coordinated affinity chromatography and isoelectric focusing. A method of studying structural and functional heterogeneity of enzymes has been developed and tested on chymotrypsin. The enzyme, prepared from single mouse pancreata, has been fractionated with respect to function and charge content by a combination of affinity chromatography and isoelectric focusing. By comparing chymotrypsin isolated from isogenic strains, chymotrypsinogen of strains A/Sn and NZB was found to be genetically heterogeneous, thus not revealed as different chymotrypsin forms of a single zymogen. Chymotrypsinogen originating from two loci was investigated, and structural and functional differences of the corresponding enzymes were determined. At both loci, structural allelomorphism was indicated. At one locus, the structural heterogeneity was also found to be reflected in functional heterogeneity of the corresponding enzymes. By mating the two strains and fractionating the enzyme of the cross, the differences were shown to be inherited.


Genetic study of pancreatic proteinase in mice (Mus musculus): linkage of the Prt-2 locus on chromosome 8. The Prt-2 locus is linked with Es-1 and Es-2 loci on chromosome 8 (linkage group XVIII). Recombination frequencies were 8.2% between Es-1 and Es-2 12.7% between Es-1 and Prt-2, and 4.5% between Es-2 and Prt-2. From these data, the map position of Prt-2 has been estimated on chromosome 8. The Prt-1 and Prt-3 loci, which are linked very closely on the same chromosome, were not determined.


Gene action in fish of tetraploid origin. V. Cellular RNA and protein content and enzyme activities in cyprinid, clupeoid, and salmonoid species. The ratio of cellular RNA and protein content is about 1:1 between phylogenetically diploid and tetraploid species of the teleost family Cyprinidae, but is roughly in proportion to ploidy in species of the teleost order Isospondyli. Enzyme activities do not unequivocally comply with this scheme. These findings are discussed in view of the hypothesis that a regulatory mechanism which reduces genic activity has evolved in the tetraploid cyprinids but not in the tetraploid species of the order Isopondyli.


An investigation of the products of 53 gene loci in three species of wild Canidae: Canis lupus, Canis latrans, and Canis familiaris. This article describes an investigation of inter- and intraspecific variation in three small populations of wild Canidae-wolf, coyote, and dingo. The products of 53 gene loci were examined. Very little interspecies variation was observed, but the level of intraspecific variation was compatible with that found in man.


Demonstration of a new mammalian isoleucine catabolic pathway yielding an Rseries of metabolites. 1. Normal human urine contains small amounts (less than 4 mg/g of creatinine) of 2-ethylhydracrylic acid, formed, we believe, by a previously undisclosed endogenous catabolic pathway for the oxidation of a newly described series of R metabolites of isoleucine. 2. Urinary excretion of 2-ethylhydracrylic acid is variably increased in defects of isoleucine oxidation at distal steps in the catabolic pathway (3-oxoacyl-CoA thiolase deficiency and methylmalonyl-CoA mutase deficiency) and is diminished when proximal steps of the oxidative pathway are blocked as in branched-chain oxo acid decarboxylase deficiency ('maple-syrup-urine' disease). 3. Precursors of R-pathway metabolites [R(-)-2-methylbutyrate and 2-ethylacrylate ] lead to increased 2-ethylhydracrylate excretion in the mammal(rat, rabbit and dog); the corresponding S metabolites [S(+)-2-methylbutyric acid and tiglic acid ], when given in equimolar amounts, have little effect on its excretion, suggesting that little or no interconversion between S and R metabolites occurs in vivo. 4. Studies with 2H-labelled precursors indicate that conversion of R 2-methylbutyrate into 2-ethylhydracrylic acid occurs by a direct pathway (apparently via 2-ethylacrylic acid). 5. The further oxidation of 2-ethylhydracrylic acid to ethylmalonic acid was demonstrated, and may be analogous to S-metabolite oxidation via methyl malonate. 6. Valine metabolites do not interact with the R=isoleucine pathway under the conditions of these experiments in vivo.


Radioactive labelling of ribosomal proteins with reductive alkylation and its use in studying ribosome-cytosol interactions. Mouse brain ribosomes were radioactively labelled by a cell-free reductive alkylation reaction with NaBH4 and [14C]formaldehyde. The radioactivity was largely associated with ribosomal proteins, but little, if any, of the rRNA was radioactive after the alkylation procedure. Both ribosomal structural proteins and loosely associated components were successfully labelled by this procedure. The sedimentation properties of the ribosomes were unaltered and their ability to carry out poly(U)-directed protein synthesis, although decreased, was largely retained. Incubation of 14C-labelled ribosomes with brain cytosol resulted in a 17% loss of radioactivity, although treatment of the ribosomes with 1.0 m-KCl to remove the loosely associated factors rendered the ribonucleoprotein particles resistant to cytosol effects. The ribosome-cytosol interactions did not appear to be related to an exchange process, since the released radioactivity was largely degraded to acid-soluble material. In addition, the incubation of native ribosomes with brain cytosol resulted in an almost complete loss in the ability of the ribosomes to participate in cell-free protein synthesis.


The effect of calcium ions on testosterone production in Leydig cells from rat testis. Leydig-cell suspensions, prepared from rat testes, were incubated with different amounts of Ca2+ with and without added luteinizing hormone. The basal testosterone production in the absence of luteinizing hormone was unaffected by the Ca2+ concentration in the incubation medium. The luteinizing hormone-stimulated testosterone production, however, was progressively decreased in the absence of Ca2+ to one-third of that with 2.50 mM-Ca2+. This decrease in luteinizing hormone-stimulated testosterone production was independent of the different concentrations of luteinizing hormone (0-10mug/ml) used and could be restored by the addition of Ca2+ to the incubation medium. The restoration of the stimulation was achieved within 30 min after the addition of Ca2+ to the medium. Activation of cyclic AMP-dependent protein kinase by luteinizing hormone was not decreased by omission of Ca2+ from the incubation medium, suggesting that Ca2+ may be involved in steroidogenesis at a stage beyond the luteinizing hormone receptor-adenylate cyclase-protein kinase system.


Total plasmalogens and O-(acylalkylglycerophosphoryl) ethanolamine from labelled hexadecanol and palmitate during hypoxia and anoxia in rat heart. By the use of the Langendorff technique, surviving isolated rat hearts were perfused with [1-14 C] palmitate, [1-14C] hexadecanol or [1-14C,1-3H] hexadecanol under normal or anoxic conditions. After perfusion for 30min with either precursor, when oxygenated or in an hypoxic condition, or when 1mM-KCN was included in the system, the heart tissues showed no significant chemical changes in their content of total lipids, total phospholipids or total ethanolamine-containing phospholipids. Changes were observed in the ratio of alkyl-to alk-1-enyl-glycerophosphorylethanolamine in the tissue perfused with N2+CO1 plus CN-. A slight increase from 4.0+/-0.3 to 4.9+/-0.2% in alkyl derivatives and a decrease from 11.2+/-0.4 to 9.4+/-0.3% in alk-1-enyl derivatives was observed. The incorporation of the [14C] palmitate and the [14C] hexadecanol into the recovered phospholipids and plasmalogens was severely decreased in the tissues perfused with CN-: in the hypoxic state only a mild inhibition was observed compared with the oxygenated systems. Considerable 3H from [1-14C, 1-3H] hexadecanol was retained (25-35%) in the alk-1-enylether chains of plasmalogens under both the oxygenated conditions and with CN-, suggesting that the same mechanism of incorporation is operational at high or low O2 concentrations. The results are consistent with an O2-dependent, CN-sensitive step in the biosynthesis of plasmalogens in the rat heart.


Intranuclear maturation pathways of rat liver ribosomal ribonucleic acids. The maturation of pre-rRNA (precursor to rRNA)in liver nuclei is studied by agar/ureagel electrophoresis, kinetics of labelling in vivo with [14C] orotate and electron-microscopic observation of secondary structure of RNA molecules. (1) Processing starts from primary pre-rRNA molecules with average mol. wt. 4.6X10(6)(45S) containing the segments of both 28S and 18S rRNA. These molecules form a heterogeneous peak on electrophoresis. The 28S rRNA segment is homogeneous in its secondary structure. However, the large transcribed spacer segment (presumably at the 5'-end) is heterogeneous in size and secondary structure. A minor early labelled RNA component with mol.wt. about 5.8X10(6) is reproducibly found, but its role as a pre-rRNA species remains to be determined. (2) The following intermediate pre-rRNA species are identified: 3.25X10(6) mol.wt.(41S), a precursor common to both mature rRNA species ; 2.60X10(6)(36S) and 2.15X10(6)(32S) precursors to 28S rRNA; 1.05X10(6) (21S) precursor to 18S rRNA. The pre-rRNA molecules in rat liver are identical in size and secondary structure with those observed in other mammalian cells. These results suggest that the endonuclease-cleavage sites along the pre-rRNA chain are identical in all mammalian cells. (3) Labelling kinetics and the simultaneous existence of both 36S and 21S pre-rRNA reveal that processing of primary pre-rRNA in adult rat liver occurs simultaneously by at least two major pathways: (i) 45S leads to 41S leads to 32S+21S leads to 28S+18S rRNA and (ii) 45S leads to 41S leads to 36S+18S leads to 32S leads to 28S rRNA. The two pathways differ by the temporal sequence of endonuclease attack along the 41 S pre-rRNA chain. A minor fraction (mol.wt.2.9X10(6), 39S) is identified as most likely originating by a direct split of 28S rRNA from 45S pre-rRNA. These results show that in liver considerable flexibility exists in the order of cleavage of pre-rRNA molecules during processing.


Re-activation of the peptidyltransferase centre of rabbit reticulocyte ribosomes after inactivation by exposure to low concentrations of magnesium ion. 1. The larger subrivosomal particles of rabbit reticulocytes retained full activity in the puromycin reaction and in poly(U)-directed polyphenylalanine synthesis after 4h at 0 degrees C when buffered 0.5M-NH4Cl/10-30mM-MgCl2 was the solvent. 2. Activity in the puromycin reaction was diminished to approx 10% after 15-30 min at 0 degrees C when the concentration of MgCl2 was lowered to 2mM. 3. Activity was not restored when the concentration of MgCl2 was raised from 2mM to 10-30 mM at 0 degrees C. However, activity was recovered as measured by both assay systems when the ribosome fraction was heated to 37 degrees C at the higher concentrations of MgCl2. 4. Recovery of activity was noted during the course of the polyphenylalanine synthesis in 50 mM-KCl/5mM-MgCl2/25mM-Tris/HCl, pH 7.6, at 37 degrees C. Re-activation was slow at 20 degrees C and below. 5. No more than about 5% of the protein moiety of the subparticle was lost in 0.5M-NH4Cl on decreasing MgCl2 concentration from 10mM to 2mM. No proteins were detected in the supernatant fractions by gel electrophoresis after ribosomes were separated by differential centrifugation. The supernatant fraction was not essential for the recovery of activity. However, at higher (e.g. 1M) concentrations of NH4Cl, proteins were split from the subparticle. 6. The loss and regain of activity found on lowering and restoring the concentration of MgCl2 at 0.5M-NH4Cl appears to arise from a conformational change that does not seem to be associated with a loss and regain of particular proteins. 7. A 2% decrease in E260 was noticed when the concentration of Mg2+ was restored, and the change in the spectrum indicated a net increase of approx. 100A-U base-pairs per subribosomal particle. 8. When the concentration of Mg2+ was restored, S20,W of the subparticle remained at 52+/- 1S until the sample was incubated at 37 degrees C when S20,W increased to 56 +/- 1S compared with the value of 58 +/- 1S for the subparticle as originally isolated.


Reassembly of the peptidyltransferase centre of larger subparticles of rabbit reticulocyte ribosomes from a core-particle and split-protein fraction. We report the reconstruction, from a core-particle and split-protein fraction, of the larger subribosomal particle of rabbit reticulocytes. The reassembled particle was active in polyphenylalanine synthesis and in the puromycin reaction. The core-particles and split-protein fractions were obtained by treatment of the larger subparticle with salt solutions containing NH4+ and Mg2+ in the molar ratio 40:1 over the range 2.25-2.75 M-NH4Cl/56-69mM-MgCl2 at 0 degrees C. This treatment led to the loss of about eight proteins (approx. 17% of the protein moiety), which were found wholly or largely in the split-protein fraction as shown by two-dimensional gel electrophoresis. The core particle retained 5S rRNA and had much decreased (no more than 10% of control) ability to function in the puromycin reaction or in poly (U)-directed polyphenylalanine synthesis. Activity was recovered when the recombined core-particle and split-protein fractions were dialysed overnight at 4 degrees C against 0.3M-NH4Cl/15mM-MgCl2/1mM-dithiothreitol/15% (v/v) glycerol/20mM-Tris/HCl, pH 7.6, and then heated for 1 h at 37 degreesEES C. The recovery was 40-80% of the original activity. Raising the concentration of MgCL2 to 300 mM in 2.5 M-NH4CL led to the removal of seven rather than eight proteins, and the core particle remained active in the puromycin reaction. We infer that the protein retained by raising the concentration of Mg2+ is an essential component of the peptidyltransferase centre of the ribosome.


Degradation of hepatic zinc-thionein after parenteral zinc administration. A low-molecular-weight protein, zinc-thionein, a metallothionein, was implicated as having a regulatory function in zinc metabolism. The half-life (t 1/2) of hepatic zinc-thionein was determined by pulse-labelling with either L-[35S] cystine and/or 65Zn. In two experiments with L-[35S]cystine, the t 1/2 of zinc-thionein was 18h and 19h. Most of the soluble 35S-labelled hepatic proteins had a t 1/2 of 4 days. The t 1/2 of zinc-thionein calculated by using 65Zn was 20h. The close similarity between the calculated and measured t 1/2 values for zinc-thionein suggests that release of Zn2+ from zinc-thionein probably occurs simultaneously with degradation of the protein moiety.


Tryptophan concentrations in rat brain. Failure to correlate with free serum tryptophan or its ratio to the sum of other serum neutral amino acids. Groups of rats were deprived of food overnight and then given free access to diets designed to raise (carbohydrate) or lower (carbohydrate and large neutral amino acids) brain tryptophan concentrations. Similar diets were supplemented with 40% fat and fed to other groups. All animals were killed 2h after food presentation. Sera from animals fed carbohydrate plus fat contained 2.5 times as much free tryptophan concentrations did not differ. Similarly, sera from rats fed on carbohydrate, large neutral amino acids, and 40% fat contained 5 times as much free tryptophan as those from rats given this meal without fat, but brain tryptophan concentrations increased by only 26%. Correlations were made between brain tryptophan and (1) free serum tryptophan, (2) the ratio of free serum tryptophan to the sum of the other large neutral amino acids in serum that compete with it for uptake into the brain, (3) total serum tryptophan or (4) the ratio of total serum tryptophan to the sum of its circulating competitors. The r values for correlations (3) and (4) (i.e. those involving total serum tryptophan) were appreciably higher than those for correlations (1) and (2). Brain tyrosine concentrations also were found to correlate well with the ratio of serum tyrosine to the sum of its competitors. Competition for uptake into the brain among large neutral amino acids (represented here by serum ratios) thus appears to determine the changes in the brain concentrations of these amino acids under physiological conditions(i.e. after food consumption). Total, not free, serum tryptophan is the relevant index for predicting brain tryptophan concentrations.


A quantitative study of pinocytosis and lysosome function in experimentally induced lysosomal storage. The highly pinocytic epithelial cells of the visceral yolk sac from 17.5-day rat conceptuses were used as a model in which to induce engorgement of the vacuolar system by direct accumulation of substances that are not hydrolysed by lysosomal enzymes. The ultra-structural appearances of these cells in pregnant animals that 24-48h before had received intraperitoneal injections of Triton WR-1339, polyvinylpyrrolidone, dextran or sucrose revealed gross abnormalities that were confined to the vacuolar system; in comparison with normal tissue the number, and in some cases the size, of vacuoles was increased, leading to close packing within the apical cytoplasm and distortion of the normal rounded shape. By culturing yolk sacs in vitro, rates of ingestion of 125I-labelled polyvinylpyrrolidone and of 125I-labelled bovine serum albumin were determined, together with the rate of digestion of the labelled protein. The rates of exocytosis of 125I-labelled polyvinylpyrrolidone and of lysosomal enzymes were also determined. No significant differences between normal and highly vacuolated tissues were found. Apparently marked vacuolation of these cells by these agents is without significant effect on pinocytosis, exocytosis or intralysosomal proteolysis.


Latency of some glycosidases of rat liver lysosomes. The latency of the alpha-glucosidase activity of intact rat liver lysosomes was studied by using four substrates (glycogen, maltose, p-nitrophenyl, alpha-glucoside, alpha-fluoroglucoside) at a range of substrate concentrations. The results indicate that the entire lysosome population is impermeable to glycogen and maltose, but a proportion of lysosomes are permeable to alpha-fluoroglucoside and a still higher proportion permeable to p-nitrophenyl alpha-glucoside. Incubation at 37 degrees C in an osmotically protected buffer of of pH 5.0 caused lysosomes to become permeable to previously impermeant substrates and ultimately to release their alpha-glucosidase into the medium. The latencies of lysosomal beta-glucosidase and beta-galactosidase were examined by using p-nitrophenyl beta-glucoside and beta-galactoside as substrates. The results indicate permeability properties to these substrates similar to that to p-nitrophenyl alpha-glucoside. On incubation in an osmotically protected buffer of pH 5, lysosomes progressively released their beta-galactosidase in soluble form, but beta-glucosidase remained attached to sedimentable material. Lysosomal beta-glucosidase was inhibited by 0.1% Triton X-100; alpha-glucosidase and beta-galactosidase were not inhibited.


Platelet-aggregating activity of type I and type III collagens from human aorta and chicken skin. Human or chicken type III collagen dissolved in 0.1 M-acetic acid was much more potent than type I collagen at inducing platelet aggregation. After incubation in 0.38M-Na2HPO4 to promote fibrillogenesis, the platelet-aggregating activity of both collagen types increased, and type I was then virtually equiactive with type III. Preincubation in cell-free plasma increased the activity of chicken but not that of human collagen. The platelet-aggregating activity of type III collagen did not appear to depend on the integrity of the intrachain disulphide bonds.


Developmental changes in purine phosphoribosyltransferases in human and rat tissues. 1. The hypoxanthine/guanine and adenine phosphoribosyltransferase activities in a wide variety of human tissues were studied during their growth and development from foetal life onward. A wide range of activities develop after birth, with especially high values in the central nervous system and testes. 2. Postnatal development of hypoxanthine/guanine phosphoribosyltransferase was also defined in the rat. Although there were increases in the central nervous system and testes, there was also a rise in activity in the liver, which was less marked in man. 3. A sensitive radiochemical assay method, using dTTP to inhibit 5'-nucleotidase activity, suitable for tissue extracts, was developed. 4. No definite evidence of the existence of tissue-specific isoenzymes of hypoxanthine/guanine or adenine phosphoribosyltransferase was found. Hypoxanthine/guanine phosphoribosyltransferase in testes, however, had a significantly different thermal-denaturation rate constant. 5. The findings are discussed in an attempt to relate activity of hypoxanthine/guanine phosphoribosyltransferase to biological function. Growth as well as some developmental changes appear to be related to increase in the activity of this enzyme.


Effect of L-azetidine-2-carboxylic acid on glycosylations of collagen in chick-embryo tendon cells. The glycosylations of hydroxylysine during collagen biosynthesis in isolated chick-embryo tendon cells were studied by using pulse-chase labelling experiments with [14C]-lysine. The hydroxylation of lysine and the glycosylations of hydroxylysine continued after a 5 min pulse label for up to about 10 min during the chase period. These data differ from those obtained previously in isolated chick-embryo cartilage cells, in which, after a similar 5 min pulse label, these reactions continued during the chase period for up to about 20 min. The collagen synthesized by the isolated chick-embryo tendon cells differed markedly from the type I collagen of adult tissues in its degree of hydroxylation of lysine residues and glycosylations of hydroxylysine residues. When the isolated tendon cells were incubated in the presence of L-azetidine-2-carboxylic acid, the degree of glycosylations of hydroxylysine during the first 10 min of the chase period was identical with that in cells incubated without thcarboxylic acid for at least 60 min, whereas no additional glycosylations took place in the control cells after the 10 min time-point. As a consequence, the collagen synthesized in the presence of this compound contained more carbohydrate than did the collagen synthesized by the control cells. Additional experiments indicated that azetidine-2-carboxylic acid did not increase the collagen glycosyltransferase activities in the tendon cells or the rate of glycosylation reactions when added directly to the enzyme incubation mixture. Control experiments with colchicine indicated that the delay in the rate of collagen secretion, which was observed in the presence of azetidine-2-carboxylic acid, did not in itself affect the degree of glycosylations of collagen. The results thus suggest that the increased glycosylations were due to inhibition of the collagen triple-helix formation, which is known to occur in the presence of azetidine-2-carboxylic acid.


Effect of maternal ethanol consumption on foetal and neonatal rat hepatic protein synthesis. Effects of maternal ethanol consumption were investigated on the rates of protein synthehsis by livers of foetal and neonatal rats both in vivo and in vitro, and on the activities of enzymes involved in protein synthesis and degradation. The rates of general protein synthesis by ribosomes in vitro studied by measuring the incorporation of [14C]leucine into ribosomal protein showed that maternal ethanol consumption resulted in an inhibition of the rates of protein synthesis by both foetal and neonatal livers from the ethanol-fed group. The rates of incorporation of intravenously injected [14C]leucine into hepatic proteins were also significantly lower in the foetal, neonatal and adult livers from the ethanol-fed group. Incubation of adult-rat liver slices with ethanol resulted in an inhibition of the incorporation of [14C]leucine into hepatic proteins; however, this effect was not observed in the foetal liver slices. This effect of externally added ethanol was at least partially prevented by the addition of pyrazole to the adult liver slices. Pyrazole addition to foetal liver slices was without significant effect on the rates of protein synthesis. Cross-mixing experiments showed that the capacity of both hepatic ribosomes and pH5 enzyme fractions to synthesize proteins was decreased in the foetal liver from the ethanol-fed group. Maternal ethanol consumption resulted in a decrease in hepatic total RNA content, RNA/DNA ratio and ribosomal protein content in the foetal liver. Foetal hepatic DNA content was not significantly affected. Ethanol consumption resulted in a significant decrease in proteolytic activity and the activity of tryptophan oxygenase in the foetal, neonatal and adult livers. It is possible that the mechanisms of inhibition of protein synthesis observed here in the foetal liver after maternal ethanol consumption may be responsible for at least some of the changes observed in 'foetal alcohol syndrome'.


A proposed sequence of hormones controlling the induction of luteal 20alpha-hydroxy steroid dehydrogenase and progesterone withdrawal in the late-pregnant rat. 1. The previously reported induction of luteal 20alpha-hydroxy steroid dehydrogenase by administration of aminoglutethimide to late-pregnant rats was shown to be unaffected by prior removal of the foetuses. Aminoglutethimide therefore does not act via the foetuses in this context. 2. The ability of injected oestrogen to prevent the above induction was lost by delaying the injection for 12h after aminoglutethimide, although the increase in enzyme activity begins only after 24h. 3. Induction of 20alpha-hydroxy steroid dehydrogenase by foetoplacental removal on day 18 of pregnancy was inhibited by human choriogonadotropin, lutropin (luteinizing hormone) and pregnant-mare serum gonadotropin, but not by somatotropin (growth hormone), thyrotropin or follitropin (follicle-stimulating hormone) 4. Indomethacin blocked the normal induction of 20alpha-hydroxy steroid dehydrogenase in late pregnancy and that caused by aminoglutethimide. It partially blocked that caused by human choriogonadotropin given on days 19-20 and that caused by 2-bromo-alpha-ergocryptine on days 5-6, but failed to block that caused by human choriogonadotropin on days 15-16 or by foetoplacental removal on day 18 of pregnancy. 5. These findings, and the control of progesterone synthesis in late pregnancy, are interpreted in terms of a sequence of hormonal or enzymic syntheses, each of which is inhibited by the product of the preceding synthesis. Several experiments were conducted to explain the natural induction of 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD) at the end of pregnancy in the rat. It has previously been showh that aminoglutethimide induces luteal 20alpha-HSD in  late-pregnancy in the absence of fetuses.  When estrone is injected within 6 hours of the administration of aminoglutethimide, the induction of 20alpha-HSD is prevented. However, this effect is lost if estrone is injected after 6 hours, even though 20alpha-HSD activity does not begin  until 24 hours after administration of aminoglutethimide. Human chorionic gonadotropin, luteinizing hormone, and pregnant mare serum gonadotropin, but not somatotropin, thyrotropin or follicle stimulating hormone, inhibited induction of  20alpha-HSD by fetoplacental removal on Day 18 of pregnancy. Normal and aminoglutethimide induction of 20alpha-HSD was blocked by indomethacin.  Indomethacin partially blocked the induction of 20alpha-HSD by human chorionic gonadotropin administered on Days 19-20 of pregnancy and that induced by 2-bromo-alpha-ergocryptine on Days 5-6, but had no effect on the induction of enzyme  activity by human chorionic gonadotropin on Days 15-16 or by fetoplacental removal on Day 18 of pregnancy. The results are explained in regards to a sequence of hormonal or enzymic syntheses, each of which is inhibited by the result of the  preceding synthesis.


The glycerol phosphate, dihydroxyacetone phosphate and monoacylglycerol pathways of glycerolipid synthesis in rat adipose-tissue homogenates. 1. Fat-free homogenates from the epididymal fat-pads of rats were used to measure the rate of palmitate esterification with different substrates. The effectiveness of the acyl acceptors decreased in the order glycerol phosphate, dihydroxyacetone phosphate, 2-octadecenyl-glycerol and 2-hexadecylglycerol. 2. Glycerol phosphate and dihydroxyacetone phosphate inhibited their rates of esterification in a mutually competitive manner. 3. The esterification of glycerol phosphate was also inhibited in a partially competitive manner by 2-octadecenylglycerol and to a lesser extent by 2-hexadecylglycerol. However, glycerol phosphate did not inhibit the esterification of 2-octadecenylglycerol. 4. The esterification of dihydroxyacetone phosphate and 2-hexadecylglycerol was more sensitive to inhibition by clofenapate than was that of glycerol phosphate. Norfenfluramine was more effective in inhibiting the esterification of 2-hexadecylglycerol than that of glycerol phosphate or dihydroxyacetone phosphate. 5 It is concluded that rat adipose tissue can synthesize glycerolipids by three independent routes.


The effects of diet on the esterification of glycerol phosphate, dihydroxyacetone phosphate and 2-hexadecylglycerol by homogenates of rat adipose tissue. 1. Male rats were fed for 5 weeks after weaning on a diet containing (by weight) 59% of starch or on diets that contained 39% of starch and 20% of either sucrose, beef tallow or corn oil. 2. The rats fed on the beef tallow consumed more energy than did the rats fed on the starch and sucrose diets. The rats fed on the corn oil drank less water than did the other groups of rats. 3. There were no significant differences between the four groups in terms of body-weight gain, epididymal-fat-pad weight and in the size, number and triacylglycerol content of the adipocytes in the fat-pads. 4. There was a significant correlation (P less than 0.001) between the activities of glycerol phosphate acyltransferase and monoacylglycerol acyltransferase in individual rats. Both of these activities were highest in the group fed on the high-starch diet and both correlated with the consumption of glucose by individual rats in the four groups. 5. The percentage of glycerol phosphate converted into diacylglycerol and triacylglycerol was positively correlated with the mean diameters, surface area and triacylglycerol content of the adipocytes for individual rats and was greates in the sucrose-fed rats. 6. The specific activity of dihydroxyacetone phosphate acyltransferase was highest in the rats fed on beef tallow. This activity was positively correlated with the energy intake for all dietary groups over the 5-week feeding period. 7. The results are discussed in terms of the functions of the three routes of glycerolipid synthesis in adipose tissue.


Uptake and effects of copper in rat liver mitochondria. The rate and extent of Cu2+ uptake by rat liver mitochondria was measured under various conditions. 1. The uptake is both greater and faster without an energy supply. 2. The uptake, when occuring in ionic media, has a biphasic character, that is it always slows down after an initial burst, and then re-accelerates. 3. Uptake of Cu2+ in the presence of energy initiates K+ uptake from K+-containing media with accompanying swelling and respiratory stimulation. Depending on the amounts of Cu2+ added and the K+ concentration, an inhibition of respiration later ensues. 4. Chelation of the Cu2+ by substrates (notably glutamate) decreases the effects. 5. Prior exposure to Cu2+ decreases or prevents energy-dependent Ca2+ uptake.


Orotate decreases the inhibitory effect of ethanol on galactose elimination in the perfused rat liver. 1. The galactose-elimination rate in perfused livers from starved rats was decreased in the presence of ethanol (2-28mM) to one-third of the control values. Orotate injections partly reversed the effect of ethanol, so that the galactose-elimination rate was about two-thirds of the control values. Orotate alone had no effect on the galactose-elimination rate. 2. Ethanol increased [galactose 1-phosphate] and [UDP-galactose], and decreased (UDP-glucose] and [UTP], both with and without orotate. Orotate increased [UTP], [UDP-galactose], both with and without ethanol. The increase of [galactose 1-phosphate] in the presence of ethanol was inhibited by orotate. Orotate alone had no appreciable effect on [galactose 1-phosphate]. 3. Both the effect of ethanol and that of orotate on the galactose-elimination rate can be accounted for by assuming inhibition of galactokinase by galactose 1-phosphate with Ki about 0.2mM, the inhibition being either non-competitive or uncompetitive. 4. The primary effect of ethanol seems to be inhibition of UDP-glucose epimerase (EC 5.1.3.2), followed by accumulation of UDP-galactose, trapping of UDP-glucose and increase of [galactose 1-phosphate]. Orotate decreased the effect of ethanol, probably by increasing [UDP-glucose].


Microbiological degradation of bile acids. Nitrogenous hexahydroindane derivatives formed from cholic acid by Streptomyces rubescens. The metabolism of cholic acid (I) by Streptomyces rubescens was investigated. This organism effected ring A cleavage, side-chain shortening and amide bond formation and gave the following metabolites: (4R)-4-[4alpha-(2-carboxyethyl)-3aalpha-hexahydro-7abeta-methyl-5-oxoindan-1 beta-yl]valeric acid (IIa) and its mono-amide (valeramide) (IIb); and 2,3,4,6, 6abeta,7,8,9,9aalpha,9bbeta-decahydro-6abeta-methyl-1H-cyclopenta[f]quinoline-3,7-dione(IIIe)and its homologues with the beta-oriented side chains, valeric acid, valeramide, butanone and propionic acid, in the place of the oxo group at C-7, i.e.compounds (IIIa), (IIIb), (IIIc) and (IIId) respectively. All the nitrogenous metabolites were new compounds, and their structures were established by partial synthesis except for the metabolite (IIIc). The mechanism of formation of these metabolites is considered. A degradative pathway of cholic acid (I) into the metabolites is also tentatively proposed.


Ribonucleic acid synthesis in rabbit erythroid cells. Kinetic studies on the synthesis of RNA in mature bone-marrow erythroid cells from rabbits were made by measuring the incorporation of [2-3H]adenosine into the ATP pool and RNA over periods up to 8h. By use of equations to fit the pool specific radioactivity and an equation using the same type of pool to generate the rate of linear DNA synthesis, good agreement between the pool parameters is found, provided that the ATP pool is measured in whole cell extracts, and assuming that the dATP and ATP pools equilibrate rapidly. RNA-synthesis rates were measured by using curve fits to equations developed by using the pool specific-radioactivity curves. The rate of synthesis of poly(A)-containing RNA varied in three experiments from 90 to 220mol/min per cell, with half-life of nuclear processing of 12-22 min with a mean of 16 min. Ribosomal RNA is synthesized at a rate of 70-200 mol/min per cell with an average half-life of nuclear processing of 37 min for the 18S RNA and 214 min for the 28S RNA. When the stable rRNA components are subtracted from the nRNA synthesis, the rate of nRNA synthesis is between 2 and 6fg/min per cell with an average half-life of degradation of 27 min. The rate of synthesis of poly(A)-containing RNA is 1.5-3.5% of the RNA-synthesis rates. These rates are compared with the RNA-synthesis rates found in L cells and concentrations of globin mRNA found in various erythroid-cell preparations.


Microbiological degradation of bile acids. The conjugation of a certain cholic acid metabolite with amino acids in Corynebacterium equi. 1. (4R)-4[4alpha-(2-Carboxyethyl)-3aalpha-hexahydro-7abeta-methyl-5-oxoindan-1beta-yl]valeric acid (II) could not be utilized by Arthrobacter simplex, even though the acid was one of the metabolites formed from cholic acid (I) by this organism. Therefore the further degradation of the acid (II) by Corynebacterium equi was investigated to identify the intermediates involved in the cholic acid degradation. 2. The organism, cultured in a medium containing the acid (II) as the sole source of carbon, produced unexpected metabolites, the conjugates of this original acid (II) with amino acids or their derivatives, although the yield was very low. These new metabolites were isolated and identified by chemical synthesis as the Na-((4R)-4-[4alpha-(2-carboxyethyl)-3a alpha-hexahydro-7a beta-methyl-5-oxoindan-1 beta-yl]-valeryl) derivatives of L-alanine, glutamic acid, O-acetylhomoserine and glutamine, i.e. compounds (IIIa), (IIIb), (IIId) respectively. 3. The possibility that the bacterial synthetic reaction observed in the acid (II) metabolism with C. equi is analogous to peptide conjugation known in both animals and higher plants is discussed. A possible mechanism for this bacterial conjugation is also considered.