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Biomedical subjects

H C Powell

Publications and source records attributed to H C Powell.

At least 37 records · Page 2Linked to original sources

Transgenic models to assess the pathogenic actions of cytokines in the central nervous system.

In order to better understand the actions of proinflammatory cytokines in the mammalian CNS, a transgenic approach was employed in which the expression of IL-6, IL-3 or TNF-alpha was targeted to astrocytes in the intact CNS of mice. Transgenic mice exhibited distinct chronic-progressive neurological disorders with neurodegeneration and cognitive decline due to IL-6 expression, macrophage/microglial-mediated primary demyelination with motor impairment due to IL-3 expression and lymphocytic meningoencephalomyelitis with paralysis induced by TNF-alpha expression. Thus, expression of specific cytokines alone in the intact CNS results in unique neuropathological alterations and functional impairments, thereby directly implicating these mediators in the pathogenesis of CNS disease.

Animals↗

Mast cell interactions with the nervous system: relationship to mechanisms of disease.

In summary, mast cell interactions in the nervous system are relevant to both physiological processes (i.e. reproduction) and pathologic states (i.e. inflammatory demyelination, painful disorders, toxic and metabolic disease, and tumor angiogenesis). Their physiologic roles may contribute to gender-related vulnerability to inflammatory disease and may modulate sensitivity to pain. Mast cells are universally involved in tissue repair and they release and respond to trophic factors such as NGF. These cells also produce and react to cytokines, and thus appear to play a role in tissue degeneration as well as repair. In certain neurological diseases, i.e. multiple sclerosis and Guillain-Barré syndrome, the ability of mast cell proteases to degrade specific myelin proteins suggests that these cells are agents, rather than bystanders, in the demyelinative process. Even more intriguing is their recently identified capacity to process bacterial antigen as efficiently as activated macrophages, suggesting that a more critical role than previously suspected might be considered for mast cells in CNS and PNS demyelination. In experimental metabolic disorders such as galactose intoxication and thiamine deficiency, mast cells appear to play a pathogenic role. Thus, in galactose intoxication, altered BNB vascular permeability occurs in conjunction with mast cell proliferation and degranulation, while in thiamine deficiency, increased histamine levels have been reported in the rat thalamus (79) and are associated with cell death and proliferation as well as mast cell degranulation (Powell and Langlais, unpublished observations). Structural interactions between mast cells and a variety of other cells have been observed, as well as close approximation of mast cells to nerve endings in tissues in which mast cells are especially active. Due to their paracrine nature, mast cells can modulate events in their microenvironment through explosive degranulation, piecemeal degranulation, or "transgranulation" as they insert granules into neighboring cells. Lastly, these cells play specific roles in reparative processes, e.g. angiogenesis, and are active in neoplastic states, including von Recklinghausen's disease (neurofibromatosis). Their involvement may have been underestimated in neuropathological studies, to date, by a reliance on staining techniques that are inadequate for identifying degranulated and therefore activated mast cells (4). More exacting histochemical and immunostaining procedures will help to fully realize the extent of their participation in physiological and pathological processes.

Animals↗

Cloning of monoclonal autoantibodies to epitopes of oxidized lipoproteins from apolipoprotein E-deficient mice. Demonstration of epitopes of oxidized low density lipoprotein in human plasma.

Many reactive products may be formed when LDL undergoes lipid peroxidation, which in turn can react with lipids, apoproteins, and proteins, generating immunogenic neoepitopes. Autoantibodies recognizing model epitopes of oxidized low density lipoprotein, such as malondialdehydelysine, occur in plasma and in atherosclerotic lesions of humans and animals. Because apo E-deficient mice develop particularly high titers of such autoantibodies, we used their spleens to clone 13 monoclonal antibodies to various epitopes of oxidized LDL ("E0 antibodies"). Binding and competitive RIAs demonstrated significant differences in fine specificity even between E0 antibodies initially selected for binding to the same screening antigen. For example, some E0 antibodies selected for binding to malondialdehyde-LDL also recognized copper oxidized LDL, acrolein-LDL, or LDL modified by arachidonic or linoleic acid oxidation products. Circulating IgG and IgM autoantibodies binding to copper-oxidized LDL, 4-hydroxynonenal-LDL, acrolein-LDL, and LDL modified with arachidonic or linoleic acid oxidation products were found in apo E-deficient mice, suggesting that the respective antigens are formed in vivo. Epitopes recognized by some of the E0 monoclonal antibodies were also found on human circulating LDL. Each of the E0 monoclonal antibodies immunostained rabbit and human atherosclerotic lesions, and some of them yielded distinct staining patterns in advanced lesions. Together, this suggests that the natural monoclonal antibodies recognize different epitopes of complex structures formed during oxidation of lipoproteins, or epitopes formed independently at different lesion sites. Our data demonstrate that a profound immunological response to a large number of different epitopes of oxidized lipoproteins occurs in vivo. The availability of "natural" monoclonal autoantibodies should facilitate the identification of specific epitopes inducing this response.

Acrolein↗

Macrophage/microglial-mediated primary demyelination and motor disease induced by the central nervous system production of interleukin-3 in transgenic mice.

Activated macrophage/microglia may mediate tissue injury in a variety of CNS disorders. To examine this, transgenic mice were developed in which the expression of a macrophage/microglia activation cytokine, interleukin-3 (IL-3), was targeted to astrocytes using a murine glial fibrillary acidic protein fusion gene. Transgenic mice with low levels of IL-3 expression developed from 5 mo of age, a progressive motor disorder characterized at onset by impaired rota-rod performance. In symptomatic transgenic mice, multi-focal, plaque-like white matter lesions were present in cerebellum and brain stem. Lesions showed extensive primary demyelination and remyelination in association with the accumulation of large numbers of proliferating and activated foamy macrophage/microglial cells. Many of these cells also contained intracisternal crystalline pole-like inclusions similar to those seen in human patients with multiple sclerosis. Mast cells were also identified while lymphocytes were rarely, if at all present. Thus, chronic CNS production of low levels of IL-3 promotes the recruitment, proliferation and activation of macrophage/microglial cells in white matter regions with consequent primary demyelination and motor disease. This transgenic model exhibits many of the features of human inflammatory demyelinating diseases including multiple sclerosis and HIV leukoencephalopathy.

Animals↗

Axonal viability and the persistence of thermal hyperalgesia after partial freeze lesions of nerve.

Partial freeze injuries of rat sciatic nerve have been used to investigate the relationship between the amount of tissue injured and the magnitude and duration of the resulting thermal hyperalgesia. Complete freeze injury of peripheral nerve produces temporary anesthesia to peripheral stimuli and is a useful neurolytic technique. The present study examined the hypothesis that incomplete nerve lesions, in which some fibers survive while others undergo Wallerian (axonal) degeneration, lead to development of thermal hyperalgesia. In this study, performed on rats, one sciatic nerve was frozen with a cryoprobe (-60 degrees C) for periods ranging from 2 to 60 s. The behavioral response to thermal stimulation of the hind footpad was tested pre- and postoperatively at days 3, 5, 7, 9, 13, 15, 19, 22, 26, and 32. In separate groups of animals with similar lesions, nerves were removed and processed for electron microscopy. Light and electron microscopy were used to determine the nature of injury to nerve fibers and its extent. There was a direct relationship between the duration of the freeze lesion and: (1) the number of nerve fibers injured; and (2) the magnitude of the resulting Wallerian degeneration. Following partial sciatic nerve injury with axonal degeneration, hyperalgesia developed within several days, peaked at approximately 1 week postinjury, and resolved during the next several weeks. Both the magnitude of the hyperalgesic response and its persistence were directly related to the duration of the freeze lesion, except in those animals in which all nerve fibers were damaged. These animals were anesthetic to thermal stimulation of the experimental footpad. This relationship helps explain the pathogenesis of painful syndromes associated with failed cryoneurolysis and may be useful in itself as a model of neuropathic pain.

Animals↗

Myasthenia gravis-like syndrome induced by expression of interferon gamma in the neuromuscular junction.

Abnormal humoral responses toward motor end plate constituents in muscle induce myasthenia gravis (MG). To study the etiology of this disease, and whether it could be induced by host defense molecules, we examined the consequences of interferon (IFN) gamma production within the neuromuscular junction of transgenic mice. The transgenic mice exhibited gradually increasing muscular weakness, flaccid paralysis, and functional disruption of the neuromuscular junction that was reversed after administration of an inhibitor of acetylcholinesterase, features which are strikingly similar to human MG. Furthermore, histological examination revealed infiltration of mononuclear cells and autoantibody deposition at motor end plates. Immunoprecipitation analysis indicated that a previously unidentified 87-kD target antigen was recognized by sera from transgenic mice and also by sera from the majority of human MG patients studied. These results suggest that expression of IFN-gamma at motor end plates provokes an autoimmune humoral response, similar to human MG, thus linking the expression of this factor with development of this disease.

Animals↗

Evolution of neuropathologic abnormalities associated with blood-brain barrier breakdown in transgenic mice expressing interleukin-6 in astrocytes.

As both astrocytes and cytokines modulate the permeability of cerebral endothelial cells, transgenic animal models which overexpress cytokines, such as interleukin-6 (IL-6), may provide insight into the neuropathological consequences of increased BBB permeability. In this study, a GFAP-IL6 transgenic mouse model and horseradish peroxidase (HRP) were used to investigate BBB permeability and associated neuropathologic changes. In the cerebellum of control mice, the BBB developed between postnatal days 7 and 14. In transgenic mice, the BBB never developed and extensive breakdown was evident in both high- and low-expressor animals by 1 month after birth. Vascular proliferation was apparent from birth in association with development and retention of normal cerebellar architecture until 3 and 6 months in high- and low-expressor animals, respectively. At these times, a leptomeningeal inflammatory infiltrate, vacuolated astrocytic foot processes and endothelial abnormalities were apparent in the cerebellum. At 6 months in high-expressor and 12 months in low-expressor animals, parenchymal inflammation, gliosis, spongiform change, axonal degeneration and macrophage accumulation were evident. The findings suggest that increased production of IL-6 can influence the development and physiologic function of the BBB as well as contribute to parenchymal central nervous system injury.

Animals↗

Toxic neuropathies.

Current publications describe neurotoxic effects of metals, pharmaceutical products, and environmental, biological and experimental substances. Agents such as lead remain the object of new epidemiological methods to identify victims. The use of methylcobalamin to ameliorate acrylamide toxicity is being explored. Antiarrhythmic and antilipidemic drugs continue to be associated with neuropathic effects but appear to be amenable to adjustments in dosage. To help control the neurotoxic effects of the chemotherapeutic drugs taxol and cisplatin analogs of adrenocorticotropic hormone and neurotrophic factors are being used. A new immunosuppressant, FK 506, has replaced cyclosporin to facilitate organ transplantation, but unwanted effects, including peripheral neuropathy, have been documented in some patients. In experimental studies, FK 506 has been reported to accelerate the rate of nerve regeneration. Botulinum toxin for the treatment of localized spastic disorders is a useful therapy, but training and supervision has been recommended by the American Academy of Neurology. Experimental toxins, such as imminodipropionitrile, continue to provide useful insights into physiologic mechanisms, including the axonal transport of cytoskeletal components.

Drug-Related Side Effects and Adverse Reactions↗

Mast cell degranulation and blood-nerve barrier permeability in rat sciatic nerve after 7 days of hyperglycemia.

The association between hyperglycemia and altered blood-nerve barrier permeability was examined after 7 days of experimental diabetes. In nerves of rats fed a diet of 40% galactose, permeability to [14C]mannitol [13.43 +/- 2.47 x 10(-5) (SD) ml.s-1.g dry wt-1] and water content [3.43 +/- 0.24 (SD) mg/mg dry wt] were significantly increased compared with control (9.24 +/- 2.09 x 10(-5) ml.s-1.g dry wt-1 and 2.15 +/- 0.28 mg/mg dry wt) and streptozotocin-diabetic animals (8.43 +/- 2.94 x 10(-5) ml.s.-1.g dry wt-1 and 2.35 +/- 0.56 mg/mg dry wt). Electron microscopy revealed significant increases in the number of degranulating perivascular mast cells and in an index of vasoconstriction in galactose-treated rats (3.8 +/- 1.6 and 0.160 +/- 0.062, respectively) compared with control (0.5 +/- 0.8 and 0.072 +/- 0.017, respectively) and diabetic animals (1.4 +/- 1.7 and 0.083 +/- 0.033, respectively). The data are consistent with a role for mast cells in permeability changes occurring after only 7 days of galactose intoxication.

Animals↗

Neuropathology of propionic acidemia: a report of two patients with basal ganglia lesions.

Propionic acidemia is a rare genetic disorder of amino acid metabolism caused by deficient activity of propionyl coenzyme A carboxylase. Neuropathologic changes previously reported in infants have been white-matter vacuolization or spongiosis. In children who survive beyond infancy, abnormalities have been found primarily in the basal ganglia. We report neuropathologic findings in two patients with propionic acidemia diagnosed in infancy who survived 35 months and 9 years, respectively. Examination of the brain of the 35-month-old boy showed vascular and parenchymal mineralization, focal pallor and spongy change, and foci of acute neuronal injury. These changes were similar to those previously described. The 9-year-old girl was in good metabolic control when she died, and presented a neuropathologic picture not previously described. She was found at autopsy to have acute hemorrhagic lesions in the caudate, putamen, and globus pallidus bilaterally and in the left ventral thalamus. There was focal neuronal loss, but no acute hypoxic/ischemic neuronal injury. Vascular proliferation and swollen endothelial cells were seen in the basal ganglia, thalamus, and substantia nigra, but not in other regions of the brain. Electron microscopy showed swelling of endothelial cells with viable adjacent brain parenchyma. The endothelial changes suggest a breakdown of the blood-brain barrier.

Basal Ganglia Diseases↗

Inhibition of macrophage chemotaxis and peripheral nerve regeneration in normal and hyperglycemic rats by the aldose reductase inhibitor Tolrestat.

This study examined the effect of Tolrestat, an inhibitor of aldose reductase, on the regenerative capacity and macrophage chemotactic property of crush-injured sciatic nerve in normal and galactose-fed rats. Galactose intoxication reduced the incidence of regeneration but did not alter the regeneration distance or the injury-induced increase in vasoactive intestinal polypeptide content of dorsal root ganglia. Tolrestat improved the incidence of regeneration in galactose-fed rats but significantly (P < 0.05) reduced the distance of nerve regeneration in both control and galactose-fed rats. Galactose intoxication enhanced the ability of homogenates of nerve undergoing Wallerian degeneration to attract macrophages, whereas chemotaxis toward nerve homogenates from Tolrestat-treated rats was absent. Tolrestat, but not the structurally dissimilar aldose reductase inhibitors Ponalrestat and Sorbinil, exhibited a reversible, dose-dependent inhibition of macrophage chemotaxis induced by polyinosinic acid. These data suggest that exaggerated sugar metabolism by aldose reductase may restrict the ability of nerve to initiate regeneration but is not responsible for the reduced distance of nerve regeneration or attenuated increase in vasoactive intestinal polypeptide production that occur after crush injury of diabetic rats. Inhibition of macrophage responses to chemotactic signals by Tolrestat may impede regeneration and other reparative mechanisms.

Aldehyde Reductase↗

Decreased endoneurial fluid electrolytes in normal rat sciatic nerve after aldose reductase inhibition.

The role of the enzyme aldose reductase in nerve homeostasis was examined by treating rats with an aldose reductase inhibitor. Female Sprague-Dawley rats were treated with Ponalrestat (25 mg/kg/day) or with excipient alone for 4 to 12 weeks before examining electrophysiologic function, endoneurial fluid electrolyte concentrations, nerve polyol levels, water content and (Na+,K+)-ATPase activity. Sorbitol, the product of glucose metabolism by aldose reductase, was detected in all nerves from control animals, whereas it was below detection limits in 7 of 11 nerves from Ponalrestat-treated rats. Ponalrestat treatment reduced endoneurial fluid sodium and chloride concentrations by 25% and 37%, respectively (both P < 0.001). No differences in nerve water content, conduction velocity, or ATPase activities were detected. These data, and previous studies demonstrating that increased flux through aldose reductase causes the accumulation of endoneurial electrolytes, suggest a role for this enzyme in modulation of the endoneurial microenvironment. However, short-term inhibition of aldose reductase does not appear to affect nerve function. Thus, our findings do not elicit concerns regarding the use of aldose reductase inhibitors in the treatment of clinical diabetic neuropathy.

Adenosine Triphosphatases↗

The role of focal nerve ischemia and Wallerian degeneration in peripheral nerve injury producing hyperesthesia.

BACKGROUND: A new model of pain associated with an experimental peripheral mononeuropathy has stimulated interest in mechanisms of pain and their structural correlates in peripheral nerve, the site of the experimental lesion. METHODS: The pathology of the neuropathy was studied and the results correlated with alterations in nerve blood flow and with the behavioral response to heat applied to the foot. The focal neuropathy was created by loosely tying several ligatures around rat sciatic nerve, which produces hyperesthesia in the ligated limb in 3-5 days. The neuropathology was striking with epineurial and endoneurial vascular stasis, edema, and extensive nerve fiber injury in the ligated segment noted at 1 week after ligation. RESULTS: Nerve blood flow was reduced significantly in the ligated segment during the development of the hyperesthesia response, suggesting that changes in nerve blood flow caused by the ligature compression of the epineurial vessels contributes to the nerve fiber injury and pathophysiology of the model. To further test this hypothesis, the epineurial vasculature was removed from 1-cm lengths of rat sciatic nerve, which reduces nerve blood flow by 58%, and by ligation of the ipsilateral femoral artery, which focally reduces nerve blood flow by 70%, and the behavioral response to heating of the paw was evaluated at 1 week. Crush injury was used as a positive control creating Wallerian degeneration without a substantial reduction in nerve blood flow. CONCLUSIONS: The results suggest that ischemia is an important initial pathogenic mechanism in the hyperesthesia associated with the loose ligature pain model, in so far as it produces Wallerian degeneration and axonal injury. Modest degrees of ischemia producing only demyelination did not produce significant hyperesthesia.

Animals↗

Schwann cell injury is attenuated by aldose reductase inhibition in galactose intoxication.

Four months of galactose intoxication induces a dose-dependent osmotic imbalance of the nerve microenvironment characterized by polyol, water, and electrolyte accumulation. Recently, dose-dependent cellular lesions have been described in the sciatic nerves of galactose-intoxicated rats. The present study was designed to demonstrate that the cell injury and endoneurial osmotic imbalance in galactose intoxication are dependent on the subsequent metabolism of galactose by the polyol pathway. Three groups of age-matched, female Sprague-Dawley rats were fed a control diet or diets containing complete micronutrient supplements with 40% galactose or 40% galactose and 0.04% Ponalrestat, an aldose reductase inhibitor (ARI). After 4 to 5 months, sciatic nerves were analyzed for polyol, water and endoneurial electrolyte content and processed for light and electron microscopic examination. Ponalrestat prevented myo-inositol depletion and accumulations of dulcitol, water and endoneurial fluid electrolytes. Axonal size-frequency histograms revealed that Ponalrestat attenuated the shift toward smaller fibers and the decrease in mean axonal diameter seen in untreated galactose-fed rats. Electron microscopic examination showed widespread reactive and degenerative changes in Schwann cells of galactose-intoxicated rats that culminated in cytoplasmic disintegration. Quantitative electron microscopy revealed that ARI treatment significantly reduced the incidence of abnormal Schwann cells. These observations indicate that the osmotic imbalance and cell injury seen in galactose intoxication are dependent on the metabolism of galactose by the polyol pathway.

Aldehyde Reductase↗

Relative neural toxicity of local anesthetics.

In rat sciatic nerve, relative neural toxicity and relative motor nerve conduction blockade were assessed for two amide-linked local anesthetics (etidocaine and lidocaine) and two ester-linked local anesthetics (chloroprocaine and procaine). As measures of neural toxicity, nerve fiber injury and edema were assayed by light microscopic examination of nerve tissue sampled 2 days after perineural (next to the sciatic nerve) injection of various concentrations of the local anesthetics. Both nerve injury and edema increased with concentration of local anesthetics, but injury was frequently present in nerve fascicles with little or no edema. In parallel studies, the amplitude of the electrical activity elicited from the interosseous muscles of the foot following ipsilateral electrical stimulation at the sciatic notch was monitored for up to 15 minutes to assess the extent of motor nerve blockade. The resulting log concentration-response curves were analyzed for differences in potency. Both for injury and for conduction block, the order of decreasing potency was: etidocaine, lidocaine, chloroprocaine, procaine. These results are not consistent with the proposal that ester-linked agents are more likely than other local anesthetic agents to cause nerve injury.

Anesthetics, Local↗

Spatial distribution of nerve injury after occlusion of individual major vessels in rat sciatic nerves.

In an attempt to better understand the spatial distribution of ischemic injury secondary to occlusion of major arteries, we measured nerve blood flow (NBF) and studied morphologic changes at various levels distal to the ligature site. Arterial ligation of the femoral, internal iliac, or superior gluteal artery was preceded and followed by measurement of NBF using laser Doppler flowmetry which helped identify "watershed areas" and guided the sampling process as nerves were examined pathologically and areas of injury were identified. Femoral artery ligation produced the most severe ischemia, focally reducing NBF by 80% in the tibial nerve at a level just below the knee. Within these ischemic nerve segments there were degenerative changes of nerve fibers seen mainly in the subperineurial region. Ligation of the internal iliac artery caused an approximately 60% reduction in NBF at the upper and mid-thigh levels of the sciatic nerve which resulted only in endoneurial edema in tissue taken at this level. Following superior gluteal artery ligation. NBF was reduced by only 20% at the pelvic level of the sciatic nerve and there was neither endoneurial edema nor fiber abnormalities. This study demonstrates the watershed pattern of ischemic injury associated with single vessel ligation by correlating neuropathologic change with quantitative measures of local nerve blood flow. The data further support the concept that mild levels of ischemia cause endoneurial edema, while moderate levels of ischemia produce demyelination and severe ischemia produces Wallerian degeneration.

Animals↗