Injury-induced apoptosis of neurons in adult brain is mediated by p53-dependent and p53-independent pathways and requires Bax.

The mechanisms of injury-induced apoptosis of neurons within the CNS are not understood. We used a model of cortical injury in rat and mouse to induce retrograde neuronal apoptosis in thalamus. In this animal model, unilateral ablation of the occipital cortex causes unequivocal apoptosis of corticopetal projection neurons in the dorsal lateral geniculate nucleus (LGN) by 7 days postlesion. We tested the hypothesis that p53 and Bax regulate this retrograde neuronal apoptosis. We found, by using immunocytochemistry, that p53 accumulates in nuclei of neurons destined to undergo apoptosis. By immunoblotting, p53 levels increase ( approximately 150% of control) in nuclear-enriched fractions of the ipsilateral LGN by 5 days after occipital cortex ablation. p53 is functionally activated in nuclear fractions of the ipsilateral LGN at 5 days postlesion, as shown by DNA binding assay (approximately fourfold increase) and by immunodetection of phosphorylated p53. The levels of procaspase-3 increase at 4 days postlesion, and caspase-3 is activated prominently at 5 days postlesion. To identify whether neuronal apoptosis in the adult brain is dependent on p53 and Bax, cortical ablations were done on p53 and bax null mice. Neuronal apoptosis in the dorsal LGN is significantly attenuated (approximately 34%) in p53(-/-) mice. In lesioned p53(+/+) mice, Bax immunostaining is enhanced in the ipsilateral dorsal LGN and Bax immunoreactivity accumulates at perinuclear locations in dorsal LGN neurons. The enhancement and redistribution of Bax immunostaining is attenuated in lesioned p53(-/-) mice. Neuronal apoptosis in the dorsal LGN is blocked completely in bax(-/-) mice. We conclude that neuronal apoptosis in the adult thalamus after cortical injury requires Bax and is modulated by p53.

Projection neurons and interneurons in the lateral geniculate nucleus undergo distinct forms of degeneration ranging from retrograde and transsynaptic apoptosis to transient atrophy after cortical ablation in rat.

The cytological responses of thalamic interneurons to selective degeneration of thalamocortical projection neurons after cortical damage in the adult brain are poorly understood. We used a unilateral neocortical lesion model (occipital cortex ablation) in the adult rat to test the hypothesis that interneurons and projection neurons in the lateral geniculate nucleus undergo distinct forms of degeneration. In situ nuclear DNA fragmentation in neurons in the lateral geniculate occurs maximally at 7 days postlesion. Geniculocortical projection neurons that are identified by the retrograde tracer Fluorogold die primarily with a morphology of endstage apoptosis prominent at 7 days postlesion. In contrast, interneurons, identified by their particular nuclear ultrastructure and by glutamic acid decarboxylase immunoreactivity, undergo an atrophic vacuolar pathology starting early during the period of projection neuron death and peaking after the projection neuron death is complete. This degeneration of interneurons is transient, because these neurons exhibit structural recovery and their numbers are not changed significantly postlesion. A rare subset of interneurons (less than one in 100 interneurons and less than one in 100 apoptotic cells) undergoes apoptosis concurrently with the projection neurons. We conclude that different types of neurons within the same thalamic nucleus respond differently to focal cortical target deprivation. Unlike the apoptosis-prone projection neurons, most interneurons undergo transient transsynaptic atrophy and recovery rather than cell death. Nevertheless, a small subset of lateral geniculate interneurons undergoes transsynaptic apoptosis in response to projection neuron apoptosis. The pathological responses of thalamic neurons to cortical trauma vary depending on cell type.

Occipital cortex ablation in adult rat causes retrograde neuronal death in the lateral geniculate nucleus that resembles apoptosis.

The mechanisms of retrograde neurodegeneration following axotomy and target deprivation in the adult central nervous system remain poorly understood. We used a unilateral occipital cortex ablation model in adult rats to test the hypothesis that retrograde neurodegeneration in the dorsal lateral geniculate nucleus resembles apoptosis. Using the retrograde tracer Fluorogold, combined with nuclear dyes or the terminal transferase-mediated deoxyuridine triphosphate-biotin nick end labeling method for detecting nuclear DNA fragmentation, apoptotic geniculocortical projection neurons were identified at approximately. six to seven days postlesion. Degeneration of dorsal lateral geniculate neurons was characterized by aberrant accumulation of perikaryal non-phosphorylated neurofilaments and, ultrastructurally, by early vacuolation and subsequent swelling of dendrites. Ultrastructural alterations in the perikaryon of dying dorsal lateral geniculate neurons included the classic chromatolytic response, with redistribution of the rough endoplasmic reticulum and dispersion of free ribosomes followed by fragmentation of the rough endoplasmic reticulum, as well as dilatation and vesiculation of the Golgi, and accumulation of intact mitochondria. Subcellular alterations evolved into classic apoptotic changes, including progressive cytoplasmic and nuclear condensation with chromatin compaction into uniformly large round clumps, while the morphological integrity of mitochondria was preserved until late in the progression of neuronal death. Cytoplasmic and then nuclear fragments budded into the surrounding neuropil and were engulfed by oligodendrocytes. We conclude that the retrograde neurodegeneration of geniculocortical neurons in adult brain results in neuronal death which has a phenotype that closely resembles apoptosis. The morphological changes that occur during this process progress from chromatolysis through consecutive stages associated with apoptosis.

Simultaneous bilateral central retinal vein occlusion associated with anticardiolipin antibodies in leukemia.

PURPOSE: To present a dramatic case of simultaneous bilateral central retinal vein occlusion associated with leukemia and anticardiolipin autoantibodies. METHODS: Interventional case report. Clinical examination, fluorescein angiography, B-mode ultrasonography, and laboratory serologies were performed on a 65-year-old man with chronic myelogenous leukemia who presented with sudden onset of decreased vision in his right eye. RESULTS: Fundus examination disclosed a nonischemic central retinal vein occlusion. Over the next 3 weeks, a nonperfused central retinal vein occlusion developed in both eyes and subsequent neovascular glaucoma developed in his left eye. Throughout this time, his white blood cell and platelet counts remained normal and his serum viscosity remained low, but anticardiolipin antibodies were increased. CONCLUSION: This case reveals the occurrence of simultaneous bilateral central retinal vein occlusion associated with anticardiolipin antibodies in leukemia and suggests an additional mechanism other than hyperviscosity for bilateral central retinal vein occlusions in leukemic patients.

Sickle cell disease presenting with extensive peri-macular arteriolar occlusions in a nine-year-old boy.

PURPOSE: To report a dramatic occlusive event of the macula surrounding the foveal avascular zone, causing severe and permanent loss of vision in a child with sickle cell disease. METHODS: Case report. A nine-year-old boy with SS hemoglobinopathy and oculocutaneous albinism developed acute unilateral loss of vision. RESULTS: Ophthalmoscopy revealed a pale, milky white, thickened retinal lesion centered on the fovea in the right eye as well as foveal hypoplasia in the left eye. The presence of macular malformation associated with oculocutaneous albinism precluded formation of a cherry-red spot. Fluorescein angiography of the right eye demonstrated extensive occlusions of the arterioles surrounding the foveal avascular zone. The presence of occlusions surrounding the fovea from multiple directions suggested the possibility of central retinal artery occlusion with migration of microemboli downstream. CONCLUSION: The patient, the youngest case reported, developed an irreversible macular infarction that was not improved by an exchange erythrocyte transfusion. He was placed on a long-term monthly transfusion protocol to protect his unaffected eye.

Neurodegeneration in excitotoxicity, global cerebral ischemia, and target deprivation: A perspective on the contributions of apoptosis and necrosis.

In the human brain and spinal cord, neurons degenerate after acute insults (e.g., stroke, cardiac arrest, trauma) and during progressive, adult-onset diseases [e.g., amyotrophic lateral sclerosis, Alzheimer's disease]. Glutamate receptor-mediated excitotoxicity has been implicated in all of these neurological conditions. Nevertheless, effective approaches to prevent or limit neuronal damage in these disorders remain elusive, primarily because of an incomplete understanding of the mechanisms of neuronal death in in vivo settings. Therefore, animal models of neurodegeneration are crucial for improving our understanding of the mechanisms of neuronal death. In this review, we evaluate experimental data on the general characteristics of cell death and, in particular, neuronal death in the central nervous system (CNS) following injury. We focus on the ongoing controversy of the contributions of apoptosis and necrosis in neurodegeneration and summarize new data from this laboratory on the classification of neuronal death using a variety of animal models of neurodegeneration in the immature or adult brain following excitotoxic injury, global cerebral ischemia, and axotomy/target deprivation. In these different models of brain injury, we determined whether the process of neuronal death has uniformly similar morphological characteristics or whether the features of neurodegeneration induced by different insults are distinct. We classified neurodegeneration in each of these models with respect to whether it resembles apoptosis, necrosis, or an intermediate form of cell death falling along an apoptosis-necrosis continuum. We found that N-methyl-D-aspartate (NMDA) receptor- and non-NMDA receptor-mediated excitotoxic injury results in neurodegeneration along an apoptosis-necrosis continuum, in which neuronal death (appearing as apoptotic, necrotic, or intermediate between the two extremes) is influenced by the degree of brain maturity and the subtype of glutamate receptor that is stimulated. Global cerebral ischemia produces neuronal death that has commonalities with excitotoxicity and target deprivation. Degeneration of selectively vulnerable populations of neurons after ischemia is morphologically nonapoptotic and is indistinguishable from NMDA receptor-mediated excitotoxic death of mature neurons. However, prominent apoptotic cell death occurs following global ischemia in neuronal groups that are interconnected with selectively vulnerable populations of neurons and also in nonneuronal cells. This apoptotic neuronal death is similar to some forms of retrograde neuronal apoptosis that occur following target deprivation. We conclude that cell death in the CNS following injury can coexist as apoptosis, necrosis, and hybrid forms along an apoptosis-necrosis continuum. These different forms of cell death have varying contributions to the neuropathology resulting from excitotoxicity, cerebral ischemia, and target deprivation/axotomy. Degeneration of different populations of cells (neurons and nonneuronal cells) may be mediated by distinct or common causal mechanisms that can temporally overlap and perhaps differ mechanistically in the rate of progression of cell death.

Astigmatism outcomes following spherical photorefractive keratectomy for myopia.

BACKGROUND: Our aim was to examine the change in astigmatism after spherical photorefractive keratectomy (PRK). These effects are essential to optimizing photoastigmatic refractive keratectomy (PARK) to correct astigmatism to within fractions of a diopter. METHODS: We retrospectively reviewed 98 eyes of 178 patients with mild to moderate myopia and cylinder < or = 1.00 diopter (D) treated with spherical PRK (VISX 20/20 STAR excimer laser system); 31 eyes had epithelium removed mechanically with a blade and 67 eyes by a laser-scrape technique. RESULTS: Refractive astigmatism was reduced by greater than 0.25 D in 27 eyes (28%); refractive astigmatism was induced by greater than 0.25 D in 31 eyes (32%); the average vector-corrected difference between an eye's astigmatism before and after surgery was 0.01 +/- 0.52 D (P = .85). Eyes with high topographic astigmatism but low refractive cylinder before PRK showed an average of 0.07 +/- -0.60 D change in refractive cylinder after PRK. Refractive astigmatism of more than 0.25 D was induced in 16 eyes (44%) that received manual removal of epithelium versus 15 eyes (24%) that received laser removal of epithelium (odds ratio 2.51, P < .01). CONCLUSIONS: When using PRK for astigmatism correction, refractive cylinder before surgery rather than topographic astigmatism may be the most appropriate method for targeting the astigmatism correction, especially when the two values are discordant. An unpredictable mild increase or decrease in astigmatism may be expected.

Hypereosinophilic sclerosing cholangitis: findings using half-Fourier magnetic resonance imaging.

Hypereosinophilic sclerosing cholangitis is a rare disease caused by eosinophilic infiltration of the gallbladder and biliary tract seen in the idiopathic hypereosinophilic syndrome. We report a 42-year-old woman who presented with symptoms of cholecystitis and obstructive cholangitis. Imaging with magnetic resonance cholangiography using a half-Fourier spinecho sequence, we were able to visualize rapidly and non-invasively a severely abnormal gallbladder, evidence of liver parenchymal inflammation, and biliary duct dilatation.

Aerosol keratopathy: a revised MacLean classification.

Case report. Chemical keratopathy has been linked to many household aerosols. We present a dramatic case of aerosol keratopathy. The finding of an unexplained epithelial or superficial stromal keratopathy must alert the physician to inquire about the use of household aerosols near the patient's face. We also propose a revised classification system based on the work of MacLean.

The effects of supercooling chemicals on myocardial ultrastructure: a transmission electron microscopy case study.

Extended ischemia results in organ infarction which limits the availability of donor hearts. Hypothermic storage extends heart preservation by effectively stopping cellular metabolism, thereby preventing toxic accumulations of metabolic wastes and depletion of energy stores. However, cell swelling as a result of ion concentration changes and cell laceration due to ice crystal growth are consequences of hypothermic ischemia. Supercooling successfully preserves hearts for an extended time without associated myocardial necrosis. The efficacies of four supercooling preservative solutions, containing hypertonic glucose, polyethylene glycol, and or winter flounder antifreeze protein, are assessed using the Langendorff isolated organ perfusion apparatus and transmission electron microscopy. Polyethylene glycol seems the most effective in preventing myocardial necrosis possibly by dehydrating, minimizing cellular ice formation, protecting against cell swelling, and functioning as an antioxidant. Hypertonic glucose seems the most effective in reducing cell swelling; it may also depress solution freezing points, bind water, adjust both intra- and extracellular osmolarities, stabilize proteins, and assist in adenosine triphosphate (ATP) production. Antifreeze protein seems to bind effectively to ice and inhibit its growth; it may also reduce membrane permeabilities to Ca2+ and K+ ions.

Apoptosis of retrogradely degenerating neurons occurs in association with the accumulation of perikaryal mitochondria and oxidative damage to the nucleus.

The mechanisms for neuronal apoptosis after axotomy and target deprivation in the adult central nervous system are poorly understood. We used a unilateral occipital cortex ablation model in the adult rat to test the hypothesis that apoptotic retrograde neurodegeneration in the dorsal lateral geniculate nucleus occurs in association with oxidative stress and mitochondrial abnormalities. Immunodetection of 8-hydroxy-2'-deoxyguanosine, a marker for oxidative injury to DNA, demonstrated that these apoptotic neurons undergo oxidative stress. Dual immunolabeling for the retrograde tracer Fluorogold to identify projection neurons and for 8-hydroxy-2'-deoxyguanosine demonstrated that apoptotic, oxidatively damaged neurons are geniculocortical projection neurons. By electron microscopy, degeneration of dorsal lateral geniculate nucleus neurons evolved in association with a transient increase in mitochondria within the perikaryon of dying neurons during the transition between chromatolysis and early apoptosis. The morphological integrity of mitochondria was preserved until late in the progression of apoptosis. The dorsal lateral geniculate nucleus ipsilateral to the cortical lesion had a transient increase in cytochrome c oxidase activity, and geniculocortical neurons at the transitional, early apoptotic stage accumulated cytochrome c oxidase activity. We conclude that axotomy-induced, retrograde neuronal apoptosis in the adult central nervous system occurs in association with the accumulation of functionally active mitochondria within the perikaryon and oxidative damage to nuclear DNA.