Aerobic exercise capacity at long-term follow-up after paediatric allogeneic haematopoietic SCT.

Peak oxygen uptake (VO2peak), a measure of aerobic exercise capacity, predicts mortality and morbidity in healthy and diseased individuals. Our aim was to determine VO2peak years after paediatric allogeneic haematopoietic SCT (HSCT) and to identify associations with baseline patient and donor characteristics, transplantation factors, pulmonary function and self-reported sports activity. In this cross-sectional, population-based study, we measured VO2peak, spirometry and diffusion capacity of the lung (DLCO) 3-10 years post HSCT. Z-scores were calculated by reference values from healthy subjects. Self-reported hours of sports activity were obtained by interview. We included 63 patients (mean age (range) 14.4 (7-24) years). HSCT patients exhibited lower mean VO2peak (-1.42 z-score, 95% confidential interval (-1.7; -1.1)) compared with healthy subjects (P<0.001). Sixteen patients (25%) had VO2peak values <-1.96 z-score. Low VO2peak was associated with reduced forced expiratory volume in 1 s (R(2)=0.11, P=0.009), reduced DLCO/VA (R(2)=0.09, P=0.01) and low physical activity (mean VO2peak z-score inactive group: -2.1 vs most active group: -1.1, P=0.02). No associations between VO2peak and diagnosis, donor type or GvHD were found. Although causes for reduced VO2peak may be multiple, our findings stress the need to focus on physical activity post HSCT to prevent lifestyle diseases and improve quality of life.

Development of a signature probe targeting the 16S-23S rRNA internal transcribed spacer of a ruminal Ruminococcus flavefaciens isolate from reindeer.

The cellulolytic Ruminococcus flavefaciens has previously been introduced into the ruminant rumen to increase microbial degradation of plant cell wall carbohydrates. The functional effect of an introduced bacterium depends on its ability to establish in the digestive tract, and signature probes can be used as a tool to track and quantify introduced strains. The purpose of this current study was to develop an oligonucleotide signature probe targeting the 16S-23S rRNA internal transcribed spacer (ITS) of a putative probiotic cellulolytic isolate (R. flavefaciens strain 8/94-32) from the rumen of reindeer (Rangifer tarandus tarandus). The 16S-23S rRNA gene ITS of three Ruminococcus strains; R. flavefaciens strain 8/94-32, R. flavefaciens FD-1 and Ruminococcus albus Ra-8, was investigated. The ITS region has been reported to vary more between closely related bacteria compared to the widely used 16S rRNA gene, and a high degree of sequence polymorphism was indeed detected between the three Ruminococcus strains studied. Based on observed sequence differences, two oligonucloetide probes, ITSRumi1 and ITSRumi2, targeting the ITS region of the R. flavefaciens isolate 8/94-32 were developed. Probe specificity was evaluated in dot blot hybridisations with R. flavefaciens isolate 8/94-32 and four other Ruminococcus-strains tested. The probe ITSRumi1 gave positive signals for the R. flavefaciens isolate 8/94-32 only, while probe ITSRumi2 gave positive signals for R. flavefaciens isolate 8/94-32 as well as for R. albus Ra-8. The result of hybridisations with the probe ITSRumi1 indicates that the probe is specific for the R. flavefaciens strain 8/94-32 amongst the four Ruminococcus-strains tested, and is promising for further studies using it as a signature probe for tracking this strain when re-introduced to the reindeer rumen.

Effects of natural winter pasture and commercial pellet on the ultrastructure of small intestinal epithelium in reindeer.

Segments of small intestine (duodenum, jejunum and ileum) from slaughtered reindeer (Rangifer tarandus tarandus) grazing natural winter pastures (n=3) and reindeer fed commercially available pellets (RF-80) in winter (n=5) were collected and immediately fixed in McDowell's fixative. Transmission electron microscopy was employed to investigate the ultrastructural features of the epithelium and lamina propria along the small intestine and to relate these to the different diets. Major differences in ultrastructural features were observed between the small intestinal enterocytes of reindeer fed the two diets. Enterocytes in reindeer fed the natural diet displayed a normal appearance with a dense cytoplasm and distinct microvilli. In contrast, reindeer fed the commercial diet showed damaged enterocytes amongst the normal cells. Abnormal changes included disintegration and loss of microvilli, cytoplasmic swelling, loss of membrane integrity and increases in the width of intercellular spaces, especially in the jejunum.

Effects of seasonal changes in food quality and food intake on the transport of sodium and butyrate across ruminal epithelium of reindeer.

Transport of 22Na and 14C-butyrate across the ruminal epithelium of captive reindeer fed a concentrate diet in summer (n=5) and in winter (n=5) and from free-ranging reindeer taken from summer (n=3) and winter pasture (n=5) was measured in vitro in Ussing chambers. Significant amounts of both Na+ and butyrate were transported across the isolated epithelium without any external driving force. The ruminal transport of Na+ and butyrate were interacting, as evidenced by both the observed amiloride-induced reduction of net butyrate-transport and by the positive correlation between net transport of butyrate and Na+. Amiloride also reduced the net transport of Na+ without significantly affecting the short-circuit current, indicating the presence of an apical Na+/H+ exchanger in the ruminal epithelium of reindeer. The captive reindeer increased the dry matter intake of a constant quality concentrate from winter to summer, but this neither affected their ruminal transport capacity nor their ruminal surface enlargement factor (SEF). Free-ranging reindeer increased their ruminal transport capacity for Na+ and butyrate from summer to winter but simultaneously reduced their ruminal SEF. The present data indicate that this food-induced increase in transport capacity was attributed to changes in the nutrient composition of the diet.

Molecular cloning of a mammalian nuclear phosphoprotein NUCKS, which serves as a substrate for Cdk1 in vivo.

We have isolated and characterized a cDNA encoding a mammalian nuclear phosphoprotein NUCKS, previously designated P1. Molecular analyses of several overlapping and full-length cDNAs from HeLa cells and rat brain revealed a protein with an apparent molecular mass of 27 kDa in both species. The deduced amino-acid sequences are highly conserved between human and rodents, but show no homology with primary structures in protein databases or with translated sequences of cDNAs in cDNA databanks. Although the protein has some features in common with the high mobility group proteins HMGI/Y, attempts to find a putative protein family by database query using both sequence alignment methods and amino-acid composition have failed. Northern blot analyses revealed that human and rat tissues contain three NUCKS transcripts varying in size from 1.5 to 6.5 kb. All human and rat tissues express the gene, but the level of transcripts varies among different tissues. Circular dichroism analysis and secondary structure predictions based on the amino-acid sequence indicate a low level of alpha helical content and substantial amounts of beta turn structures. The protein is phosphorylated in all phases of the cell cycle and exhibits mitosis-specific phosphorylation of threonine residues. Phosphopeptide mapping and back-phosphorylation experiments employing NUCKS from HeLa interphase and metaphase cells show that the protein is phosphorylated by Cdk1 during mitosis of the cell cycle.

Use of illegal drugs among mothers across racial/ethnic groups in the United States: a multi-level analysis of individual and community level influences.

OBJECTIVE: To obtain adjusted one-year prevalence estimates and compare multi-level correlates of marijuana, cocaine, crack, or heroin use among 4,678 mothers of minor children in the United States across racial/ethnic populations. METHODS: The study used publicly available data from the 1997 National Household Survey on Drug Abuse. Hierarchical generalized linear modeling (HGLM) procedures were used to test drug use variation among non-Hispanic White, non-Hispanic Black, and Hispanic mothers across US neighborhoods. RESULTS: Black mothers were 40% less likely to use illegal drugs than were White mothers, and Hispanic mothers were 72% less likely than were White mothers to use drugs, after adjusting for demographic and community covariates. Drug use was found to vary across neighborhoods for all racial/ethnic groups. This study identified a positive association between the likelihood of mothers using illegal drugs and their living in neighborhoods with a higher occurrence of drug use. CONCLUSION: To compare drug use across racial/ethnic groups, it is necessary to account for the confounding effects of covariates. Additional research is needed to determine whether non-drug using mothers chose to live in neighborhoods with a lower concentration of drug use, whether residence in these neighborhoods decreases the likelihood of mothers using drugs, or if a reciprocal relationship exists. The mechanisms that link individual drug use and neighborhood characteristics merit further investigation.

Functional anatomy of the omasum in high Arctic Svalbard reindeer (Rangifer tarandus platyrhynchus) and Norwegian reindeer (Rangifer tarandus tarandus).

The structure and fill of the omasum was investigated in summer and in winter in adult female reindeer living on the polar desert and tundra of the high Arctic archipelago of Svalbard and in sub-Arctic mountain habitats in northern Norway. The mean total mass of the omasum in non-lactating adult female Svalbard reindeer was 467 g (0.65 g per 100 g live body mass (BM)) in September and 477 g (1.03 g per 100 g BM) in April. By contrast, the mean mass of the omasum in non-lactating adult reindeer in northern Norway was 534 g (0.83 g per 100 g BM) in September but only 205 g (0.35 g per 100 g BM p < 0.05) in late March, owing to a decrease in both tissue mass and the wet mass of the contents of the organ. The mean absorptive surface of the omasum in Svalbard reindeer was 2300 cm2 in September and 2023 cm2 in April. In Norwegian reindeer, by contrast, the absorptive surface area decreased from 2201 cm2 in September to 1181 cm2 (p < 0.05) in late March. The marked seasonal decline of omasal tissue and contents in Norwegian reindeer probably results from intake of highly digestible forage plants, including lichens, in winter. Svalbard reindeer, a non-migratory sub-species, survive eating poor quality fibrous vascular plants in winter. The absence of any marked seasonal change in the mass, total absorptive surface area or filling of the omasum in Svalbard reindeer in winter despite a substantial decline in body mass presumably reflects their need to maintain maximum absorption of nutrients, including volatile fatty acids, when feeding on such poorly fermentable forage.

Chitinolytic bacteria in the minke whale forestomach.

Minke whales consume large amounts of pelagic crustaceans. Digestion of the prey is initiated by indigenous bacteria in a rumen-like forestomach system. A major structural component of the crustacean exoskeleton is chitin, the beta-1,4-linked polymer of N-acetyl-D-glucosamine. The exoskeletons appear to dissolve completely in the non-glandular forestomach. Bacteria in the forestomach fluid of six krill-eating minke whales were enumerated and isolated using an anaerobic habitat-simulating culture medium. Median viable population densities ranged between 6.0 x 10(6) and 9.9 x 10(9) bacterial cells per mL forestomach fluid. Bacterial isolates (n = 44) cultured from the forestomach fluid of one minke whale mainly resembled strains of Eubacterium (25%), Streptococcus (18%), Clostridium (14%), and Bacteroides (11%). As much as 12% of the bacterial isolates were chitinolytic, while beta-N-acetylglucosaminidase activity was demonstrated in 54% of the isolates, and utilisation of N-acetyl-D-glucosamine was observed in 73%. The chitinolytic isolates resembled strains of Bacteroides, Bacteroidaceae, Clostridium, and Streptococcus. Scanning and transmission electron microscopy of partly digested krill from the minke whale forestomach revealed bacteria close to and inside the chitinous exoskeleton. The bacterial chitinase may act on the chitinous crustacean exoskeletons, thereby allowing other bacteria access to the nutritious soft inner tissues of the prey, and thus initiating its degradation and fermentation.

Bilirubin inhibits Ca2+-dependent release of norepinephrine from permeabilized nerve terminals.

Although the well-known neurotoxic agent bilirubin can induce alterations in neuronal signaling, direct effects on neurotransmitter release have been difficult to demonstrate. In the present study we have used permeabilized nerve terminals (synaptosomes) from rat brain prelabeled with [3H]norepinephrine to examine the effects of bilirubin on transmitter release. Rat cerebrocortical synaptosomes were permeabilized with streptolysin-O (2 U/ml) in the absence or presence of bilirubin (10 microM-320 microM) and Ca2+ (100 microM), and the amount of radiolabeled transmitter released during 5 min to the medium was analysed. Low levels of bilirubin decreased Ca2+-evoked release in a dose-dependent manner, with half-maximal effect at approx 25 microM bilirubin. Higher levels of bilirubin (100-320 microM) increased [3H]norepinephrine efflux in the absence of Ca2+, suggesting that high bilirubin levels induced leakage of transmitter from vesicles. The nontoxic precursor biliverdin had no effect on Ca2+-dependent exocytosis. Our data indicate that bilirubin directly inhibits both exocytotic release and vesicular storage of brain catecholamines.

Modulation of the effect of bilirubin on protein phosphorylation by lysine-containing peptides.

Protein phosphorylation is an important mechanism for regulation of cell processes. Bilirubin inhibits phosphorylation of several peptides/proteins by a number of different kinases, and this may contribute to the toxic effects of bilirubin on cells, particularly neurons. Bilirubin binds to lysine residues on both albumin and ligandin. The ATP-binding subdomain II on protein kinases contains an invariant lysine, which might hypothetically be involved in mediation or modulation of bilirubin-inhibition of protein phosphorylation. We have studied the ability of lysine-containing peptides to modulate the effects of bilirubin, using phosphorylation of a phospholemman peptide catalyzed by protein kinase A as a model system. Addition of bilirubin (50 microM) decreased the activity of the catalytic subunit of protein kinase A by 75%. A synthetic lysine-containing decapeptide which mimicked part of subdomain II on the protein kinase family was partially able to prevent the bilirubin effects. Similar effects were not observed with two other decapeptides in which lysine had been replaced by arginine or alanine. Polylysine (100 microM) completely prevented the inhibitory effect of 50 microM bilirubin, whereas polyglutamate and polyarginine did not have this effect. Poly-D-lysine and poly-L-lysine appeared to be equivalent in their ability to prevent the bilirubin effect. These data support the notion that binding of bilirubin to lysine may play a role in the mediation and/or modulation of bilirubin neurotoxicity.

Influence of diet on the occurrence of intraepithelial microabscesses and foreign bodies in the ruminal mucosa of reindeer calves (Rangifer tarandus tarandus).

The forestomach mucosa was examined for pathological lesions in 12 healthy free-ranging reindeer calves at different seasons, and in 32 reindeer calves fed lichen (n = 3), baled grass silage (n = 24) or pelleted feed (n = 5). No gross lesions were seen. Samples for histology were taken from reticulum, omasum and four sites in rumen. In histological sections a total of 182 intraepithelial microabscesses (IEMAs) and 16 foreign body lesions (FBLs) were observed in the ruminal mucosa, while 1 IEMA was found in omasum and no lesions in reticulum. The number of animals having 0, 1-10, 11-20 and > 20 IEMAs per four ruminal sections (one from each sample site) was 19, 19, 5 and 1, respectively. None of the free-ranging or lichen-fed animals had more than 1 IEMA per four ruminal sections, while 58% (14/24) of the animals fed silage and 40% (2/5) of the animals fed pelleted feed had 2 or more IEMAs per 4 ruminal sections. FBLs occurred more frequently in animals with high numbers of IEMAs, and it is suggested that both IEMAs and FBLs are caused by plant particles penetrating the ruminal epithelium. The lesions did not seem to indicate an inferior diet nor influence the health of the animals.

Bilirubin has widespread inhibitory effects on protein phosphorylation.

The neurotoxic effects of bilirubin may involve modulation of neuronal protein phosphorylation systems. Using in vitro phosphorylation assays and a variety of protein substrates and purified protein kinases, we have studied the mechanism of bilirubin-induced inhibition of protein phosphorylation. Bilirubin was found to inhibit cAMP-dependent, cGMP-dependent, Ca(2+)- calmodulin-dependent, and Ca(2+)-phospholipid-dependent protein kinases, irrespective of substrate properties. Fifty percent inhibition occurred at bilirubin concentrations varying from 20 to 125 microM. Kinetic analysis, using the isolated catalytic subunit of cAMP-dependent kinase and a synthetic peptide substrate derived from the protein phospholemman, indicated that bilirubin (50 microM) decreased the apparent Vmax of the reaction, irrespective of whether ATP or peptide levels were varied, without significantly altering the apparent K(m) value. Thus our results indicate that bilirubin can inhibit catalytic domain(s) of protein kinases by apparent noncompetitive mechanism(s), presumably by interacting with noncatalytic domains on the enzyme. Given the key role of protein phosphorylation in cellular regulation, the widespread inhibitory effect of bilirubin on protein kinases may contribute to bilirubin neurotoxicity.

Production rates of volatile fatty acids in the minke whale (Balaenoptera acutorostrata) forestomach.

Minke whales (Balaenoptera acutorostrata) have developed a compartmentalized stomach system, which includes a non-glandular forestomach containing high concentrations of indigenous bacteria. The forestomach contents serve as microbial substrate, and samples were collected from five adult minke whales eating capelin (Mallotus villosus) and crustaceans (Thysanoessa sp.). Chemical analysis of the forestomach contents revealed that they consisted of crude protein (650 (SD 58) g/kg DM), lipid (330 (SD 77) g/kg DM) and water-soluble carbohydrates (53.3 (SD 7.3) g/kg DM). The contribution of energy from volatile fatty acids (VFA), produced by forestomach bacterial fermentation, to the total energy budget was estimated. The forestomach concentration of VFA ranged from 13.2 to 68.5 mmol/l, and the pH was 5.83 (SD 0.41). VFA pool size ranged from 72.8 to 638.1 mmol and represented from 0.169 to 2.107 kJ/kg live weight (W)0-75. Maximal recorded forestomach VFA production rate was 1694 mmol/h in one capelin-eating minke whale with 42.6 litres of forestomach fluid. Energy from VFA produced by forestomach fermentation represented 6-107 kJ/kg (W)0-75 per d, which accounts for only 0.9-16.9% of the average daily energy expenditure of minke whales. This study suggests that the bacterial fermentation in the minke whale forestomach varies, depending on the volume and the quality of substrate available, influencing fermentation rates and concentration of VFA. Due to the small relative size of the forestomach, the contribution of VFA to the daily energy requirement in minke whales would be of less importance than in ruminants even when assuming the same production rate of VFA as in a ruminant.

Effects of parental involvement on the functioning of noninstitutionalized adults with schizophrenia.

OBJECTIVES: The study described the nature and extent of parental families' involvement with adult children with schizophrenia who lived in noninstitutional settings in the community and examined the association between families' involvement and the adult children's clinical and psychosocial functioning. METHODS: A total of 193 subjects with schizophrenia were interviewed to obtain data on their contact with their families, clinical and psychosocial functioning, risk of victimization and substance use, and global functioning. RESULTS: Twenty-three percent of the subjects lived with their parental families on some time during the previous six months. Nearly two-thirds of those who did not live with their families had contact with them an average of twice a week. Subjects who did not live with their families scored significantly higher on global functioning and on measures of contact with friends, dating, number of days worked, independence from family, and stability of living situation. Those who lived with their families were less likely to have been victimized or to have used substances. Among subjects who did not live with their families, those who maintained contact scored higher than those without contact on measures of days worked and overall role functioning. CONCLUSIONS: The nature of parental families' involvement with adult children with schizophrenia varied widely. Whether families' involvement was associated with higher levels of functioning varied with the subjects' living situation.

Ruminal microbial digestion in free-living, in captive lichen-fed, and in starved reindeer (Rangifer tarandus tarandus) in winter.

In free-living (FL) reindeer eating a natural mixed winter diet dominated by lichens, captive (CF) reindeer fed pure lichens ad libitum, and CF reindeer subsequently starved for 1 day (CS1 reindeer) or 4 days (CS4 reindeer), the dominant rumen anaerobic bacteria were characterized, their population densities were estimated, and ruminal pH and volatile fatty acid concentrations were determined. In the FL reindeer, the total median viable anaerobic bacterial population ranged from 18 x 10(8) to 35 x 10(8) cells per ml of rumen fluid (n = 4), compared with 26 x 10(8) to 34 x 10(8) and 0.09 x 10(8) to 0.1 x 10(8) cells per ml of rumen fluid in CF reindeer (n = 2) and CS4 reindeer (n = 2), respectively. The median bacterial population adhering to the rumen solids ranged from 260 x 10(8) to 450 x 10(8), 21 x 10(8) to 38 x 10(8), and 0.5 x 10(8) cells per g (wet weight) of rumen solids in FL, CF, and CS4 reindeer, respectively. Although there were variations in the rumen bacterial composition among the FL reindeer (n = 4), strains of Bacteroides, Fibrobacter, Streptococcus, and Clostridium dominated in the rumen fluid. Streptococcus spp. and Clostridium spp. were the dominant bacteria in the CF reindeer (n = 2), while in the CS4 reindeer (n = 2) the dominant bacteria were Fusobacterium spp., members of the family Enterobacteriaceae, and Eubacterium spp. Transmission electron micrographs of lichen particles from the rumen of one FL reindeer, one CF reindeer, and one CS4 reindeer show bacteria resembling Bacteroides spp. adhering to the lichen particles, evidently digesting the lichen hyphae from the inside.(ABSTRACT TRUNCATED AT 250 WORDS)

Digestion of herring by indigenous bacteria in the minke whale forestomach.

Northeastern Atlantic minke whales (Balaenoptera acutorostrata) have a multichambered stomach system which includes a nonglandular forestomach resembling that of ruminants. Bacteria from the forestomachs of herring-eating whales were enumerated and isolated in an anaerobic rumen-like culture medium (M8W medium). The total viable population of anaerobic bacteria ranged from 73 x 10 to 145 x 10/ml of forestomach fluid (n = 4). Lactobacillus spp. (19.7%), Streptococcus spp. (35.9%), and Ruminococcus spp. (12.8%) were the most common of the bacterial strains (n = 117) isolated by use of M8W medium from the forestomach fluid population of two minke whales. Most of the isolates stained gram positive (93.2%), 62.4% were cocci, and all strains were strictly anaerobic. The population of lipolytic bacteria in one animal, enumerated by use of a selective lipid medium, constituted 89.7% of the viable population. The total viable population of anaerobic bacteria in freshly caught and homogenized herring (Clupea harengus) ranged from 56.7 to 95.0 cells per gram of homogenized prey (n = 3) when M8W medium was used. Pediococcus spp. (30.6%) and Aerococcus spp. (25.0%) were most common of the bacterial strains (n = 72) isolated from the homogenized herring. Most of the bacterial strains were gram positive (80.6%), and 70.8% were cocci. Unlike the forestomach bacterial population, as many as 61.1% of the strains from the herring were facultatively anaerobic. All bacterial strains isolated from the prey had phenotypic patterns different from those of strains isolated from the dominant bacterial population in the forestomach, indicating that the forestomach microbiota is indigenous. Scanning electron microscopic examinations revealed large numbers of bacteria, surrounded by a glycocalyx, attached to partly digested food particles in the forestomach. These data support the hypothesis that symbiotic microbial digestion occurs in the forestomach and that the bacteria are indigenous to minke whales.

Analysis of a Mycoplasma hominis membrane protein, P120.

The monoclonal antibody mAb 26.7D generated against a clinical isolate of Mycoplasma hominis 7488 was shown to react with a surface-exposed epitope on a 120-kDa protein (P120). The gene encoding the protein was cloned and sequenced, and the transcriptional start point was determined by primer extension analysis. The gene contained an open reading frame of 3237 bp encoding a peptide of 1079 amino acids with a deduced molecular mass of 123 kDa. A putative amino-terminal signal peptide and cleavage site for signal peptidase II were found. This suggests that the protein was synthesized as a precursor with subsequent processing to a mature lipoprotein. Surface exposure was confirmed by immunoelectron microscopy. mAb 26.7D reacted with 11 of 19 M. hominis strains. The gene was, however, present in all strains.

Seasonal changes in the adherent microflora of the rumen in high-arctic Svalbard reindeer.

Seasonal changes in bacterial colonization of the epithelial tissue were examined in the rumen of high-arctic Svalbard reindeer. Samples of tissue were collected from eight sites in the rumen of reindeer during summer and winter and bacterial colonization was examined using scanning and transmission electron microscopy. At two of these sites, colonization by adherent bacteria was estimated to cover approximately 30% of the ruminal epithelium in specimens collected from reindeer during summer. Bacteria at these sites resembled Ruminococcus sp. and were surrounded by large amounts of glycocalyx. In winter specimens, less than 10% of the epithelial surface was covered by adherent bacteria. Those bacteria that did colonize the epithelial surface were smaller and had virtually no glycocalyx on their surface. Bacteria attached to plant cell wall material in summer samples of reindeer ingesta contained large intracellular glycogen deposits, whereas feed particle-associated bacteria in ingesta collected in winter contained no intracellular glycogen. These data demonstrate that the ruminal bacterial population responds to seasonal changes in feed intake and quality. It is yet to be determined if these bacterial changes enhance the ability of Svalbard reindeer to survive in the hostile environment of the high Arctic.

Parapoxvirus papillomatosis in the muskoxen (Ovibos moschatus): genetical differences between the virus causing new outbreak in a vaccinated herd, the vaccine virus and a local orf virus.

Since 1981 a domesticated muskoxen herd had been successfully vaccinated against papillomatosis with homogenated, glutaraldehyde inactivated papilloma tissue. In the fall of 1985 a new clinical outbreak of disease occurred, affecting previously infected as well as vaccinated animals. The purification of parapox virions directly from papilloma tissue and orf scabs collected in a local sheep farm was followed by restriction endonuclease analysis of viral DNA. The morphological identity of purified virus was controlled by electron microscopy. Comparison of restriction endonuclease digests (10 different enzymes) by gel electrophoresis demonstrated that the muskoxen parapoxvirus from the new outbreak 1985 differed considerably from the 2 other isolates (muskoxen 1981 and local orf). The latter viruses demonstrated a high degree of homology, but differences were evident after digestion with the enzyme EcoRI. During metrizamide gradient purification minor bands containing morphologically intact virions were isolated in addition to the major fractions. The restriction enzyme digests indicated that the virions of the minor bands differed from those in the major bands.