A family of closely related bovine viral diarrhea virus recombinants identified in an animal suffering from mucosal disease: new insights into the development of a lethal disease in cattle

Induction of lethal mucosal disease (MD) in cattle is linked to the generation of cytopathogenic (cp) bovine viral diarrhea virus (BVDV) in animals persistently infected with a noncytopathogenic BVDV. In most cases the cp variants are generated by recombination with cellular or viral sequences. BVDV was obtained from the serum of an MD animal and propagated in tissue culture without plaque purification. Analysis of cDNA clones established from RNA of these cells showed that apparently a variety of different viral RNAs were present. Seven of the cDNA clones contained a cellular sequence coding for light chain 3 (LC3) of microtubule-associated proteins 1A and 1B. This insertion had already been found in the cp virus JaCP obtained from the same animal and isolated by plaque purification. Analysis of further plaque-purified cp viruses showed that the diseased animal contained a family of closely related cp BVDV recombinants. A set of viruses with different duplications of viral sequences in their genomes and a variety of defective viral RNAs with deletions were found that all contained the LC3* insertion. For all the recombinants the 3' recombination sites and, in all but one case, also the 5' recombination sites between cellular and viral sequence were identical. Variation between the individual deduced genome structures resulted from different duplications or deletions of viral sequences located upstream of the cellular insertion. These results suggest that within the animal a primary recombinant with a genome containing the LC3* insertion was generated. In a trimming process a set of secondary virus recombinants was generated from this hypothetical primary recombined RNA. These secondary recombinants display genome structures that represent variations of the basic scheme already present in the primary recombinant. Apparently this trimming process that finally led to an outbreak of MD lasted a long time since recombined RNA with the basic genome structure of the cp viruses could be demonstrated in samples already taken a long time before outbreak of the disease.     

Sex Differences in the Catecholamine Output of Children

Urinary excretion of catecholamines was studied in 240 normal, healthy children 12 years of age during a passive condition (film exposure) and an active condition (arithmetic test) at school. No sex differences in catecholamine excretion were found during the passive condition, whereas, during the work period, boys excreted significantly more adrenaline and noradrealine than girls. A comparison is made between catecholamine excretion levels of the children and adult subjects examined in other studies.     

Mice with mutations in Mahogunin ring finger-1 (Mgrn1) exhibit abnormal patterning of the left-right axis.

Mahogunin Ring Finger 1 (Mgrn1) encodes a RING-containing protein with ubiquitin ligase activity that has been implicated in pigment-type switching. In addition to having dark fur, mice lacking MGRN1 develop adult-onset spongy degeneration of the central nervous system and have reduced embryonic viability. Observation of complete situs inversus in a small proportion of adult Mgrn1 mutant mice suggested that embryonic lethality resulted from congenital heart defects due to defective establishment and/or maintenance of the left-right (LR) axis. Here we report that Mgrn1 is expressed in a pattern consistent with a role in LR patterning during early development and that many Mgrn1 mutant embryos show abnormal expression of asymmetrically expressed genes involved in LR patterning. A range of complex heart defects was observed in 20-25% of mid-to-late gestation Mgrn1 mutant embryos and another 20% were dead. This finding was consistent with 46-60% mortality of mutants by weaning age. Our results indicate that Mgrn1 acts early in the LR signaling cascade and is likely to provide new insight into this developmental process as Nodal expression was uncoupled from expression of other Nodal-responsive genes in Mgrn1 mutant embryos. Our work identifies a novel role for MGRN1 in embryonic patterning and suggests that the ubiquitination of MGRN1 target genes is essential for the proper establishment and/or maintenance of the LR axis.     

Partner acceptance of health department notification of HIV exposure, South Carolina

To determine the acceptability of health department notification of sex and needle-sharing partners of persons infected with human immunodeficiency virus (HIV), we administered an anonymous questionnaire to partners notified of their exposure to HIV during the previous 2 years. Of the 202 partners notified, 132 (65%) were locatable and completed the questionnaire. Only 12 (9%) thought they may have been exposed to HIV before health department notification. When the 132 partners were asked if they thought the health department did the right thing in telling them about their exposure, 87% responded "yes;" when asked if the health department should keep notifying persons exposed to HIV, 92% responded "yes." Responses were similar for homosexual-bisexual men, heterosexuals, and intravenous drug users; men and women; and whites and blacks. We conclude that health department notification is acceptable to persons exposed to HIV in this rural South Carolina district.     

Dibenzo[a,l]pyrene-induced DNA adduction, tumorigenicity, and Ki-ras oncogene mutations in strain A/J mouse lung

Dibenzo[a,l]pyrene (DB[a,l]P), an environmental polycyclic aromatic hydrocarbon, is the most potent carcinogen ever tested in mouse skin and rat mammary gland. In this study, DB[a,l]P was examined for DNA adduction, tumorigenicity, and induction of Ki-ras oncogene mutations in tumor DNA in strain A/J mouse lung. Groups of mice received a single i.p. injection of 0.3, 1.5, 3.0, or 6.0 mg/kg DB[a,l]P in tricaprylin. Following treatment, DNA adducts were measured at times between 1 and 28 days, while tumors were counted at 250 days and analyzed for the occurrence of point mutations in codons 12 and 61 of the Ki-ras oncogene. DB[a,l]P in strain A/J mouse lung induced six major and four minor DNA adducts. Maximal levels of adduction occurred between 5 and 10 days after injection followed by a gradual decrease. DB[a,l]P-DNA adducts in lung tissue were derived from both anti- and syn-11,12- dihydroxy-13,14-epoxy- 11,12,13,14-tetrahydrodibenzo[a,l]pyrene (DB[a,l]PDE) and both deoxyadenosine (dAdo) and deoxyguanosine (dGuo) residues in DNA as revealed by cochromatography. The major adduct was identified as a product of the reaction of an anti-DB[a,l]PDE with dAdo in DNA. DB[a,l]P induced significant numbers of lung adenomas in a dose- dependent manner, with the highest dose (6.0 mg/kg) yielding 16.1 adenomas/mouse. In tricaprylin-treated control animals, there were 0.67 adenomas/mouse. Based on the administered dose, DB[a,l]P was more active than other environmental carcinogens including benzo[a]pyrene. As a function of time-integrated DNA adduct levels, DB[a,l]P induced lung adenomas with about the same potency as other PAHs, suggesting that the adducts formed by DB[a,l]P are similar in carcinogenic potency to other PAHs in the strain A/J mouse lung model. Analysis of the Ki- ras mutation spectrum in DB[a,l]P-induced lung tumors revealed the predominant mutations to be G-->T transversions in the first base of codon 12, A-->G transitions in the second base of codon 12, and A-->T transversions in the second or third base of codon 61, concordant with the DNA adduct profile.      

The effect of asphyxia on superior mesenteric artery blood flow in the premature sheep fetus

BACKGROUND/PURPOSE: The aim of this study was to determine superior mesenteric artery blood flow changes during and after an asphyxial insult in utero in chronically instrumented unanaesthetised premature fetal sheep. METHODS: Fetal sheep at 0.7 gestation (103 to 104 days) underwent 25 minutes of complete umbilical cord occlusion (n = 6) or sham occlusion (n = 6). Fetal heart rate, blood pressure, superior mesenteric artery (SMA) blood flow and vascular resistance, electroencephalographic activity, and nuchal electromyographic activity were measured from 6 hours before occlusion until 3 days after occlusion. Fetal gastrointestinal tissue was taken for histological assessment. RESULTS: During occlusion, cardiovascular response was characterised by 3 phases: initial redistribution of blood flow away from the gut to maintain vital organ function, subsequently partial failure of this redistribution, and finally near terminal cardiovascular collapse with profound hypotension and gastrointestinal hypoperfusion. Postasphyxia there was a secondary period of hypoperfusion that was mediated by increased vascular resistance, not hypotension. There was no evidence of injury on standard histological assessment after 3 days of recovery. CONCLUSIONS: SMA blood flow is not only significantly reduced during asphyxia, but also for several hours after an asphyxial insult. The authors speculate that these perturbations of gastrointestinal blood flow could compromise gut wall integrity potentially leading to increased vulnerability to necrotising enterocolitis.