Experimental rabies in skunks and foxes. Pathogenesis of the spongiform lesions

The pathogenesis of rabies spongiform lesions in striped skunks (Mephitis mephitis) and red foxes (Vulpes vulpes) was studied by light and electron microscopy and peroxidase-antiperoxidase immunocytochemistry. Studies in skunks included use of several street virus variants (different antigenic profiles as tested by monoclonal antibodies) different routes of inoculation (intranasal, intracerebral and intramuscular), immunosuppression of infected skunks, different preparations of virus (brain and salivary gland suspensions and infective tissue culture fluids), and sequential development of the lesions. Foxes (Vulpes vulpes) were infected intramuscularly with a street virus isolate. Except for the group of immunosuppressed skunks, all animals that developed clinical signs of rabies had encephalitis characterized by varying degrees of mononuclear perivascular cuffing, focal gliosis, and Negri bodies. Spongiform change occurred in the neuropil of the grey matter (especially thalamus and cerebral cortex) in rabid animals from all groups, but not in controls or exposed animals that did not develop rabies. Rabies antigen (detected by peroxidase-antiperoxidase immunocytochemistry) occurred only in small amounts in many thalami; some vacuolated areas were devoid of antigen. Ultrastructurally, there was a gradation in lesions from small to large membrane-bound vacuoles in cellular processes (mainly dendrites, less frequently axons) and to large tissue spaces containing granular and/or membranous material. These studies indicate that rabies spongiform change occurs in skunks given street virus of several different antigenic profiles and challenge virus standard rabies virus and that the distribution of the lesions has remarkable similarities to those of the traditional spongiform encephalopathies. The occurrence of the lesion is not affected by the immune response, the route of inoculation of virus, the preparation (suspension of salivary gland or brain, or tissue culture fluid), or the incubation period. The paucity of antigen in many thalami suggests that incorporation of viral components into vacuolar membranes is not necessary for development of the spongiform change. The development of the lesions includes formation of small membrane-bound vacuoles in cellular processes, rapid enlargement (less than 3 days) with compression of adjacent neural tissue, and rupture resulting in the large tissue spaces readily visible by light microscopy.     

The agent of Human Granulocytic Ehrlichiosis resides in an endosomal compartment.

The composition of cytoplasmic vacuoles containing the agent of Human Granulocytic Ehrlichiosis (HGE) was studied to investigate how this pathogen exists within infected host cells. Electron microscopy demonstrated that the HGE organism resides in a membrane-bound compartment within HL-60 cells: early forms of the HGE agent have a round reticular appearance while later structures are small and dense. Vacuoles containing HGE bacteria incorporated endocytosed colloidal gold particles, suggesting that they are part of the endocytic pathway. Antibodies directed to the mannose-6-phosphate receptor labeled vacuole membranes. Antibodies to the transferrin receptor and to the lysosomal membrane glycoprotein LAMP 1 did not. Moreover, 3-(2,4-dinitroanilino)-3'-amino-N-methyldipropylamine, which normally accumulates in compartments with low pH, was not present inside these vacuoles. These results suggest that vacuoles containing the agent of HGE fail to mature into phagolysosomes. We conclude that the agent of HGE appears to enter and modify part of the endocytic pathway.     

Role for interleukin-1 (IL-1) in benzene-induced hematotoxicity: inhibition of conversion of pre-IL-1 alpha to mature cytokine in murine macrophages by hydroquinone and prevention of benzene-induced hematotoxicity in mice by IL-1 alpha

Chronic exposure of humans to benzene (BZ), a myelotoxin, causes aplastic anemia and acute leukemia. The stromal macrophage that produces interleukin-1 (IL-1), a cytokine essential for hematopoiesis, is a target of BZ's toxicity. Monocyte dysfunction and decreased IL-1 production have been shown to be involved in aplastic anemia in humans. Hydroquinone (HQ), a toxic bone marrow (BM) metabolite of BZ, causes time- and concentration-dependent inhibition of processing of the 34-Kd pre-interleukin-1 alpha (IL-1 alpha) to the 17-Kd mature cytokine in murine P388D1 macrophages and BM stromal macrophages, as measured by Western immunoblots of cell lysate proteins using a polyclonal rabbit antimurine IL-1 alpha antibody. HQ over a 10-fold concentration range had no effect on the lipopolysaccharide (LPS)-induced production of pre- IL-1 alpha precursor or on cell viability or DNA and protein synthesis. Stromal macrophages obtained from the femoral BM of C57Bl/6 mice exposed to BZ (600 or 800 mg/kg body weight) for 2 days were incapable of processing the 34-Kd pre-IL-1 alpha to the mature 17-Kd cytokine when stimulated in culture with LPS. Stromal macrophages from mice coadministered BZ and indomethacin, a prostaglandin H synthase (PHS) inhibitor that has been shown to prevent BZ-induced myelotoxic and genotoxic effects in mice when coadministered with benzene were able to convert the pre-IL-1 alpha to mature cytokine. Administration of recombinant murine IL-1 alpha (rMuIL-1 alpha) to mice before a dose of BZ that causes severe depression of BM cellularity completely prevents BM depression, most probably by bypassing the inability of the stromal macrophage in BZ-treated animals to process pre-IL-1 alpha to the mature cytokine.     

Upregulation of interleukin-1 receptors in mouse AtT-20 pituitary tumor cells following treatment with corticotropin-releasing factor.

Treatment of AtT-20 cell cultures with increasing concentrations of rat/human corticotropin-releasing factor (r/hCRF) for 24 h resulted in a dose-dependent 2-3 fold increase in specific 125I-labelled recombinant human IL-1 alpha (125I-IL-1 alpha) binding that was paralleled by a 70-80% decrease in 125I-Tyro-ovine CRF binding. Saturation analysis of 125I-IL-1 alpha binding in control and CRF-treated cultures indicated that CRF produced an increase in the density (Bmax) of IL-1 receptors without altering their affinity (KD). The CRF-induced upregulation of IL-1 receptors appears to be mediated through specific membrane receptors for CRF since the CRF receptor antagonist, alpha-helical oCRF (9-41), blocked the CRF-induced upregulation of IL-1 receptors without producing any effect on 125I-IL-1 alpha binding by itself. In summary, these data demonstrate complex interactions between CRF and IL-1 at the pituitary level and identify potential novel mechanisms for cytokines to alter neuroendocrine function.     

Successful replantation of amputated nose and auricle

Amputations of the nose and of the auricle present difficult management problems. Application of simple reattachment techniques, followed by aggressive medical therapy which consists of cooling, anticoagulation, antibiotic coverage, and multiple stab incisions in the amputated tissues, have resulted in the successful replantation of major portions of an amputated nose and an amputated auricle with satisfactory cosmetic results.     

Description of a human direct calorimeter, with a note on the energy cost of clerical work

1. A heat-sink calorimeter, suitable for the measurement of energy expenditure in human subjects over periods up to 26 h, is described. 2. The performance of the calorimeter is illustrated by a study of four normal subjects at rest or performing clerical work for a period of 7.5 h. Each condition was measured in duplicate in each subject. On the resting days the subjects were recumbent, and on the working days they were seated throughout the measurement period. Heart rate was monitored by infra-red telemetry and physical activity by an ultrasound movement detector. Urinary cortisol excretion was also measured as an indicator of stress. 3. In each subject the mean heat loss on working days was higher than that on resting days: the increase ranged from 5.1% to 16.7%, with a mean value of 10.0% (P = 0.015). There was no significant difference between resting and working days in heart rate, physical activity or urinary cortisol excretion. 4. The present study confirms that tiring clerical work has very little effect on 24 h energy expenditure.