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An insertion sequence, IS1081, in the genome of Mycobacterium bovis has been identified and sequenced. It is 1324 bp long with 15 bp inverted repeat ends and contains a large ORF. There are six copies of IS1081 in the genome of M. bovis and the element is also present in Mycobacterium tuberculosis. IS1081 is not closely related to other DNA elements described in actinomycetes but its putative transposase bears some resemblance to that of IS256 from Staphylococcus aureus. IS1081 may be useful for genetic manipulations and for developing a diagnostic test for bovine tuberculosis based on the polymerase chain reaction.

Genomic DNA was prepared from four reference strains of Mycobacterium paratuberculosis and 46 isolates of this organism from New Zealand, Australia, Canada, and Norway and also from two mycobactin-dependent "wood pigeon" strains. The DNA was characterized by restriction endonuclease analysis, both with and without DNA hybridization, with a probe specific to a repetitive DNA sequence in M. paratuberculosis. Both techniques differentiated M. paratuberculosis strains into two groups, but DNA hybridization revealed more differences between strains within the larger group. All the strains from cattle and many strains from other animals belonged to this group. The second group of nine strains included the Faroe Islands strain, all New Zealand sheep strains, and one New Zealand goat strain. Primary isolation of strains belonging to this group was difficult to achieve. DNA from acid-fast organisms harvested directly from intestinal tissues of sheep with Johne's disease was shown to have restriction and hybridization patterns identical to those of DNA obtained from M. paratuberculosis isolates cultured from the same tissues. Two Norwegian goat strains and the wood pigeon strains did not hybridize to the M. paratuberculosis probe and had restriction patterns very different from those of other M. paratuberculosis strains. The wood pigeon strains had restriction patterns very similar to those of strains of Mycobacterium avium, indicating that they should be classified as that species. The presence of two distinct groups of M. paratuberculosis strains and their predominant distribution in different host animals may be significant in management of mixed-animal farming operations.

Mycobacterium bovis was isolated from diagnostic samples from 57 cats submitted to New Zealand Animal Health Laboratories from 1974 to 1986. With six exceptions, these cats came from suburban and rural areas of New Zealand where M.bovis was also present in feral and wild animals, especially the brush-tailed possum. Tuberculous skin lesions were seen in 33 (58%) of the cats. Histologically, these lesions had some similarities to those of cat leprosy. Included in the 57 cats was a group of 12 tuberculous animals which were diagnosed in a suburban veterinary practice over a 3 month period. When these 12 M. bovis isolates were examined by DNA restriction endonuclease analysis, they were found to be identical. This evidence, together with the relatively short period during which the cases occurred, suggested that these cats were exposed to a single source of infection.

It was recently proposed that the genus Bacteroides should be restricted to Bacteroides fragilis (the type species) and closely related organisms (viz., B. caccae, B. distasonis, B. eggerthii, B. merdae, B. ovatus, B. stercoris, B. thetaiotaomicron, B. uniformis, and B. vulgatus). By contrast, the moderately saccharolytic, predominantly oral Bacteroides species, which include B. melaninogenicus, B. oralis, and related species, form a phenotypically and phylogenetically coherent group of species which differ so significantly from the emended description of the genus Bacteroides that they should not be classified in the same genus. Therefore, we formally propose that these species be reclassified in a new genus, Prevotella. The type species is Prevotella melaninogenica.

Culture of tuberculous lesions from six of 14 captive seals yielded an organism which, on the basis of biochemical and drug sensitivity tests, was identified as Mycobacterium bovis, although the organism showed a weak cording pattern and was glycerol tolerant. It was pathogenic in rabbits and guinea pigs and after passage the organism exhibited strong cord formation and was glycerol intolerant. Restriction endonuclease analysis and sodium dodecyl-sulphate polyacrylamide gel electrophoresis indicated that the organism belonged to the Mycobacterium tuberculosis complex. Restriction patterns indicated that infection in the six seals was from a single source. Western blotting with monoclonal antibody to M bovis identified antigens at 23 and 27 kDa in M bovis which were absent from the seal isolates.

1. Fatty acid binding activity associated with a 14,000-15,000 mol. wt protein was observed in the cytosolic fraction of liver, duodenum, myocardium, adipose, pectoral and gastrocnemius muscles of chickens. 2. Polyclonal antisera prepared against chicken liver fatty acid binding protein affinity for only liver FABP and a 14,000 mol. wt fatty acid binding protein in the intestine. 3. A fatty acid binding protein was not detected in chicken plasma.

Deoxyribonucleic acid (DNA) preparations from 3 reference strains of Mycobacterium paratuberculosis and from 23 isolates of M paratuberculosis obtained from cattle in New Zealand were characterized by restriction endonuclease analysis, using the enzymes BstE II, Pvu II, and Bcl I. Patterns of DNA fragments for strain 18 (one of the reference strains) differed markedly from patterns of other strains, indicating genetic differences between strain 18 and the other strains of M paratuberculosis evaluated. The other 2 reference strains (TMC 1613 and Weybridge strain 316) and all but 1 of the isolates from cattle had identical patterns with the 3 enzymes. These 2 reference strains differed from each other in their dependence on exogenous mycobactin, but this was not reflected in their restriction patterns. The single variant isolate from cattle had patterns identical to those of the other isolates, using Pvu II and Bcl I, and had only 1 fragment line difference with BstE II. Although close genetic homogeneity of cattle strains of M paratuberculosis prevented development of a typing system on the basis of restriction endonuclease analysis, the results provided a basis for genomic comparison with other closely related organisms.

An improved understanding of bile acid physiology is a prerequisite for the construction of an acceptable model for describing plasma clearance of bile acids. The aim of this study was to determine the extent of reflux unconjugated cholic acid from the liver to the plasma in subjects with normal liver function. The clearance of a bolus injection of unconjugated 14C-cholic acid was studied in four subjects using samples collected simultaneously from a peripheral and a hepatic vein. Hepatic extraction ratios calculated directly from the data were found to be essentially constant during the 20 min following injection, with average values of 0.60-0.72. This indicates that reflux did not occur significantly in the subjects investigated. These results are interpreted as favouring a model in which exchange of unconjugated 14C-cholic acid occurs between the plasma and a peripheral compartment with final removal from the plasma to the liver.

DNA restriction endonuclease analysis was used for intra-specific typing of Mycobacterium bovis isolates from 83 brush-tailed possums (Trichosurus vulpecula) obtained between 1982 and 1984 from the three major regions in New Zealand with endemic bovine tuberculosis. All the isolates were found to be genetically very similar. Differentiation of the isolates into 33 restriction types was achieved by using high-resolution electrophoresis and the combined results from separate digestions with the restriction enzymes Bst EII, Pvu II and Bcl I. The typing system was entirely reproducible. Isolates of the same type were usually found in adjacent localities and were always limited to one of the three major regions. In some cases, isolates of the same type were found in both 1982 and 1984. The phenotypic significance of the small genetic differences identified between different isolates is unknown. The typing system will be useful for monitoring the transmission of M. bovis to other species and the future spread of different M. bovis types through possum populations.

Seven daughter strains of BCG were characterized by restriction fragment analysis with the enzymes BstEII, PvuII and BclI. Comparisons of fragment patterns confirmed that BCG is correctly classified as a Mycobacterium bovis variant and suggested that the Swedish strain is most closely related to the original BCG strain.

DNA preparations from two reference (H37Ra and H37Rv) and two wild strains of Mycobacterium tuberculosis and one re-isolated strain of Mycobacterium bovis BCG were analysed using 17 restriction endonucleases. The enzyme BstEII revealed the greatest differences between strains. Electrophoretic DNA patterns from the wild M. tuberculosis strains differed from each other and from the reference strains at relatively few positions. At the highest resolution attained, patterns from the two reference strains remained indistinguishable from each other. The pattern of the M. bovis BCG strain was substantially different from, but had many bands in common with, the M. tuberculosis patterns.

Arteriohepatic dysplasia is a rare congenital syndrome in which attenuated peripheral pulmonary arteries are associated with liver impairment. The earliest indication is usually persistent cholestatic jaundice in the first few months after birth. Although this is invariably associated with pruritus, bile acid kinetic studies have not been reported in such patients. This report describes a girl with a particularly mild form of the syndrome who was never observed to be clinically or biochemically jaundiced. Bile acid studies indicated that there was a defect in bile acid excretion but not in uptake or conjugation.

Forty-three strains of Campylobacter fetus ss. fetus isolated from sheep abortions in New Zealand, and reference strains of C. fetus ss. fetus (four), C. fetus ss. venerealis (two), C. jejuni (one) and C. coli (one) were examined by restriction endonuclease analysis with the enzymes BstE II and Xho I. DNA fragment patterns of C. fetus, C. jejuni and C. coli differed strikingly from each other, but there were many similarities in the patterns obtained for all strains of C. fetus ss. fetus (47) and C. fetus ss. venerealis (three). The 43 local strains of C. fetus ss. fetus were clearly divided into four types. Two of these types produced patterns nearly identical with those of overseas reference strains.

The deliberate ingestion of carbon tetrachloride by a 48-year-old woman provided an opportunity to study the sequential biochemical changes of a severe but self-limited event of hepatocellular damage. The initial phase of cellular injury characterised by high levels of serum aspartate aminotransferase, ferritin and bile acids was followed by a period of partial cholestasis. This was indicated by a marked decrease in secondary bile acids and small rises in alkaline phosphatase and 5' nucleotidase. Improving liver function and regeneration became evident during this period and was associated with abnormal levels of gamma glutamyl transpeptidase. A late rise in serum ferritin and alphafetoprotein may represent a non-specific inflammatory reaction.

Organisms belonging to the Mycobacterium avium-M. intracellulare-M. scrofulaceum (MAIS) serocomplex were subjected to restriction endonuclease analysis (REA) with the enzymes BstEII, PvuII, and BclI. Substantial genetic heterogeneity was observed between members of an authenticated collection of the 31 serotypes. Serotypes 2 and 3 were indistinguishable, however, as were serotypes 5 and 10. No direct correlation could be made between restriction pattern and species identification. REA of serotype 2 and serotype 8 isolates from various geographic locations and animal origins showed that, within limits, the restriction pattern could be used as an index of serotype. Some isolates that were unable to be classified serologically exhibited restriction patterns identical to those of strains that were able to be classified by seroagglutination. The difficulty of interpreting much of the epidemiological data concerning MAIS organisms may be partially explained by the extent of heterogeneity observed by REA. These findings support the contention that the MAIS complex has a substantially greater degree of heterogeneity than has been revealed by traditional methods.

Serum bile acids were measured in control subjects and in 23 patients with cirrhosis or cholestasis using an enzymic method which measures total non-sulphated bile acids and 2 commercially available radioimmunoassay methods which measure cholylglycine (CG) and sulpholithocholylglycine (SLCG) respectively. All 3 methods gave abnormal values in patients with cholestasis. In cirrhosis the CG method gave abnormal values more often than the enzymic method. The SLCG method was much less sensitive. Measurement of CG in 2 postprandial samples per patient detected only one patient with normal fasting but abnormal postprandial levels. Analysis of samples by gas liquid chromatography (GLC) showed that the enzymic and CG methods gave a good estimation of total non-sulphated bile acids and cholic acid respectively. Values determined by the SLCG method were much higher than the equivalent GLC values. The CG method is faster than the enzymic method and is less expensive provided sample are analysed in sufficiently large batches.

Restriction endonuclease analysis was used to examine 70 different strains of Campylobacter fetus subsp. fetus, which were isolated from aborted sheep foetuses. The strains could be divided into seven types based on the DNA fragment patterns obtained by electrophoresis after digestion with the enzyme BstEII. With one exception, digestion with the enzyme XhoI allowed the strains to be grouped identically to that for BstEII. Antisera were made against formalized whole cells representative of each of the seven different restriction types. Analysis of these sera by tube agglutination tests using whole cells revealed five different serogroups. Examination of the 70 strains with absorbed antisera demonstrated a complex relationship between the restriction type and the external antigens of C. fetus subsp. fetus.

Brucella ovis DNA was analysed by using 11 different restriction endonucleases. The most clearly resolved DNA fragment patterns were obtained after digestion with the enzyme Hind III. When DNA preparations from 35 strains of B. ovis were digested with this enzyme, the fragment patterns appeared to be identical. The patterns obtained after Hind III digestion of DNA from one strain each of B. abortus, B. canis and B. melitensis were more similar to each other than to the B. ovis pattern.

In two groups of male rats having an average age of either 90 days or two years, guide cannulae for bilateral push-pull perfusion were implanted stereotaxically to rest upon the superficial frontal cerebral cortex. On post-operative recovery, either 1.5 or 3.0 g/kg 20% ethanol (V/V) was given by intragastric gavage to each unrestrained rat. Sequential samples of venous blood were obtained from the tail and analyzed for alcohol levels by gas chromatography. A set of push-pull perfusions of the cortical sites was carried out with an artificial CSF before gavage and at 25, 50 and 150 min after the administration of ethanol. An individual perfusion was continued for 5.0 min at a rate of 25 microliters/min. Using high performance liquid chromatography with electrochemical detection (HPLC-EC) each sample of perfusate was then assayed for its content of glutamate (Glu), aspartate (Asp), glutamine (Gln), glycine (Gly), taurine (Tau) and GABA with homoserine used as the internal standard. The results showed that the 3.0 g/kg dose of ethanol resulted in a higher level of blood ethanol in the older animals, which persisted over the 150 min time interval. Further, the 1.5 g/kg dose of ethanol administered to the older rats reduced the cortical activity of Glu and Gln relative to the younger animals. In addition, the 3.0 g/kg dose augmented the cortical efflux of Tau in the aged rats. Neither dose of ethanol affected the efflux of Asp or Gly from the perfused frontal cortex of either the young or old group, nor was the release of GABA detectable under either the control condition or following treatment with ethanol.(ABSTRACT TRUNCATED AT 250 WORDS)

A 0.2-kb DNA sequence specific to Mycobacterium paratuberculosis, the causative organism of Johne's disease, was isolated from a partial genomic library. The sequence was part of a larger repetitive DNA element and was present in strains of M. paratuberculosis from cattle, sheep, goat, deer and also a woman with Crohn's disease but not in M. paratuberculosis strain 18. The sequence was not present in strains of 19 other mycobacterial species including 31 reference serotype strains of the M. avium-M. intracellular-M. scrofulaceum (MAIS) complex, some strains of which are closely related to M. paratuberculosis. The sequence may be useful for developing a diagnostic test for Johne's disease.

Serum bile acids were measured in 28 patients with established liver disease. The peak serum level after a meal was as sensitive an index of liver disease as a combination of serum bilirubin, aspartate amino transferase, alkaline phosphatase and gamma glutamyl transpeptidase and was more often abnormal than any one of the four tests. Serum bile acid measurements may be of most value in detecting cirrhosis when the activity of disease is minimal.

Four macaque monkeys which showed excessive preference for ethyl alcohol solutions in a self-selection paradigm were used as subjects. Earlier these animals had been given intracerebroventricular (ICV) injections of human cerebrospinal fluid, which produced pharmacologically significant effects on the animals' alcohol consumption. The purpose of the present investigation was to determine whether the parenteral administration of a new anxiolytic compound, buspirone, would alter the pattern of alcohol drinking already established in the monkey. Initially the maximally selected concentration of alcohol of 12% was determined on the basis of a standard ad lib alcohol-water preference screen. Each monkey was offered 12% alcohol and water for a basal pre-injection period of 4 days. Then, on each of the next three days, either the saline control vehicle or buspirone, 1.25, 5.0 or 20.0 mg/kg, was injected intramuscularly at 930 and 1630 hours. On these days, behavioral observations were recorded before and after buspirone's administration in order to evaluate the latency as well as recovery from the drug's effect. Subsequently, a four-day post-injection, 12% alcohol-water test was conducted. Although the saline control and 1.25 mg/kg dose of buspirone were without effect on alcohol intake, both the 5.0 and 20.0 mg/kg doses of buspirone attenuated significantly the consumption of alcohol by the monkeys. This reduction in terms of absolute g/kg as well as the proportion of alcohol to water ingested was approximately 30-60% of baseline intakes. Following buspirone treatment, the amount of alcohol consumed returned essentially to previously high levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Unit dose and traditional drug distribution systems were compared in a 352-bed long-term-care facility by analyzing nursing time, medication-error rate, medication costs, and waste. Time spent by nurses in preparing, administering, charting, and other tasks associated with medications was measured with a stop-watch on four different nursing units during six-week periods before and after the nursing home began using unit dose drug distribution. Medication-error rate before and after implementation of the unit dose system was determined by patient profile audits and medication inventories. Medication costs consisted of patient billing costs (acquisition cost plus fee) and cost of medications destroyed. The unit dose system required a projected 1507.2 hours less nursing time per year. Mean medication-error rates were 8.53% and 0.97% for the traditional and unit dose systems, respectively. Potential annual savings because of decreased medication waste with the unit dose system were $2238.72. The net increase in cost for the unit dose system was estimated at $615.05 per year, or approximately $1.75 per patient. The unit dose system appears safer and more time-efficient than the traditional system, although its costs are higher.

DNA preparations from 24 New Zealand isolates, two reference strains of Mycobacterium bovis, and one reference strain each of Mycobacterium microti, Mycobacterium africanum, and Mycobacterium tuberculosis were characterized by restriction endonuclease analysis. Twenty-five restriction enzymes were investigated. The clearest differences in M. bovis patterns were obtained with the enzymes BstEII and BclI. These produced four and five different patterns, respectively, for the 24 local isolates. When the results from both enzymes were considered, seven different combinations were obtained. The patterns produced for the two reference strains of M. bovis could be distinguished from each other and also from the patterns produced for the local isolates. All patterns were reproducible and are now being used for typing M. bovis isolates. With either enzyme, the patterns produced for the M. tuberculosis, M. bovis, and M. africanum strains had many features in common, but all the M. bovis patterns were clearly more similar to each other than to the M. tuberculosis patterns. The patterns produced for the M. microti strain were markedly different from those produced for the other species. Restriction endonuclease analysis is clearly a useful method for inter- and intraspecific classifications of the tuberculosis complex.

In diabetic rats glomerular morphologic damage is exacerbated by feeding a protein-rich diet. Protein feeding alters arachidonic acid metabolism in other models of renal disease, and there is evidence that the arachidonic acid metabolite thromboxane plays a pathophysiologic role in protein-induced renal injury. In this study we evaluated the effect of high-protein feeding on renal thromboxane production, renal hemodynamics, and renal morphologic condition in rats with experimentally induced diabetes. We induced diabetes in male Sprague-Dawley rats by streptozocin administration. Rats then received high (60% casein)- or low (8% casein)-protein diets. Eight to 11 weeks later, clearance of inulin and PAH and renal blood flow were measured. Rats fed 60% casein had higher glomerular filtration rate and renal blood flow than rats fed low-protein diets. Rats fed high-protein diets had more glomerular hypercellularity, tubular hypertrophy, and arteriolar thickening than their protein-restricted counterparts. Renal production of 6-keto-PGF1a and PGE2 was not different between dietary groups. Renal thromboxane production, however, was greater in rats fed 60% protein than in rats fed 8% protein. We conclude that protein feeding stimulates renal thromboxane production and exacerbates morphologic injury in the diabetic rat. Short-term administration of the thromboxane synthetase-inhibitor UK 38,485 did not further increase glomerular filtration rate or renal blood flow in animals fed high protein. Thus thromboxane did not appear to play a role in protein-induced injury in this model by a vasoconstrictive mechanism. Other possible mechanisms by which thromboxane may contribute to the renal damage observed are discussed.