Spin trapping of azidyl and hydroxyl radicals in azide-inhibited rat brain submitochondrial particles

Succinate-driven respiration in azide-inhibited rat brain submitochondrial particles (smps) produces azidyl and hydroxyl radicals that were detected by spin trapping with 5,5'-dimethyl-1-pyrroline-N-oxide (DMPO). Production of radicals required succinate and oxygen and was eliminated by heat denaturation, which indicates that radical production is a result of respiration. The concentrations of both DMPO/.OH and DMPO/.N3 were decreased by addition of catalase to the smps, which indicates that H2O2 is involved in radical production. In the absence of azide anion, DMPO/.OH was not detected in the same system, even after five additions of succinate over a period of 24 h. It is proposed that azide inhibition of cytochrome c oxidase results in increased production of superoxide, which is efficiently converted to hydrogen peroxide by membrane-bound superoxide dismutase. Hydrogen peroxide activates endogenous peroxidase to react with azide anion forming azidyl radical, which damages the peroxidase, resulting in decreased production of azidyl radical with successive additions of succinate. Hydroxyl radical is produced from the hydrogen peroxide that is not removed by peroxidase. The increased production of superoxide in the azide-inhibited system suggests that loss of cytochrome c oxidase activity can lead to increased radical production if other proteins in the respiratory chain remain active. In the azide-inhibited system, reaction of azide anion with H2O2-activated endogenous peroxidase and spin-trapping of the resulting azidyl radical is a convenient monitor of H2O2 production.     

Assessment of colour Doppler tissue imaging using test-phantoms

An investigation has been carried out on the velocity resolution, spatial resolution and accuracy of Doppler images as part of a study into the Doppler display of cardiac tissue motion. Test-phantoms were designed to perform this work and images were captured on a computer. The characteristics of the phantom images and of the image capture process were studied. The smallest spatial detail that was observed in the Doppler image was 3 mm by 3 mm. Doppler receive gain and Doppler ensemble size both affected velocity resolution. Different target materials gave different measures for velocity resolution. This could be related to the different back-scatter intensities of the materials.     

External-bean megavoltage radiation therapy in the treatment of post-radical prostatectomy residual or recurrent tumor preliminary results

High dose, external-beam megavoltage radiation therapy was administered to 21 patients because of incomplete excision (11 patients) or palpable local recurrence (10 patients) following radical prostatectomy. Of the 21 patients 8 were alive without clinical evidence of cancer for 29 to 97 months after radiation therapy. The best survival was in the 11 patients referred because of incomplete excision. Of these 11 patients 9 were alive, 6 without cancer, and 2 were dead of intercurrent disease without evidence of cancer. Of the 10 patients with palpable local recurrence 2 were alive without evidence neoplasm, while 2 died of intercurrent disease without cancer. Acute treatment-associated symptoms occurred in 11 of the 21 patients. Generally, these symptoms were mild and responded to conservative, symptomatic management. Two serious complications were recorded.     

Immunohistochemical study of a chondroitin-6-sulfate in growth plates of broiler chickens with high and low genetic predispositions to tibial dyschondroplasia

The distribution of a chondroitin-6-sulfate (C6S) epitope, which is a biochemical marker of chondrocyte hypertrophy, was studied in the growth plates of two lines of 3-week-old broiler chickens with low and high genetic predispositions to tibial dyschondroplasia (TD). Ultrathin sections of growth plates from both groups were subjected to immunolocalization with monoclonal antibody 3-B-3(-), the epitope of which is increased on proteoglycans made by hypertrophic chondrocytes. Bound antibody was localized with colloidal gold-labeled protein A for observation with an electron microscope. The 3-B-3(-) epitope was localized in pericellular and interterritorial matrix of growth plates of both lines. In the low-TD-incidence birds, the concentration of 3-B-3(-) bound to C6S progressively increased from the proliferative zone to the hypertrophic zone. However, in the high-TD-incidence line, the epitope expression remained at a low level in all zones. The increase of the 3-B-3(-) epitope produced by maturing growth-plate chondrocytes is indicative of changes in the glycosaminoglycan chains of proteoglycans that may be important in the process of matrical calcification. Thus, failure of chondrocytes of the high-TD-incidence line to produce this change in post-translational modification of their proteoglycans could be important in the pathological process.     

Severe neuropathies in mice with targeted mutations in the ErbB3 receptor

Neuregulins and their specific receptors, members of the ErbB family of tyrosine kinases, have been implicated in the control of growth and development of Schwann cells, specialized cells that wrap around nerve axons to provide electrical insulation. Here we use gene targeting to generate mice that lack ErbB3, a high-affinity neuregulin receptor. Homozygous erbB3 mutant embryos lack Schwann-cell precursors and Schwann cells that accompany peripheral axons of sensory and motor neurons. The initial development of motor neurons and sensory neurons of dorsal root ganglia occurs as it should, but at later stages most motor neurons (79%) and sensory neurons in dorsal root ganglia (82%) undergo cell death in erbB3 mutant embryos. Degeneration of the peripheral nervous system in erbB3 mutant pups is thus much more severe than the cell death in mice that lack neurotrophins or neurotrophin receptors. We also show that ErbB3 functions in a cell-autonomous way during the development of Schwann cells, but not in the survival of sensory or motor neurons. Our results indicate that sensory and motor neurons require factors for their survival that are provided by developing Schwann cells.     

Direct arteriovenous fistulas (carotid-cavernous and vertebral-venous). Diagnosis and intervention

Carotid-cavernous and vertebral-venous fistulas are direct arteriovenous fistulas. Their symptoms range from benign to extremely severe ophthalmologic or neurologic complications. Mechanism of symptoms is mostly related to venous drainage. Therapy is widely indicated, as today's endovascular techniques--using mostly detachable balloons and coils--have a high success rate and very few complications.     

Specific triggering of the Fas signal transduction pathway in normal human keratinocytes

The epidermis is continually exposed to genotoxic injury and requires an efficient mechanism to eliminate genetically altered cells. The membrane receptor, Fas, initiates apoptosis in many cell types, including keratinocytes. Receptor cross-linking is the vital post-ligand binding step in Fas signal transduction, and we have utilized FK1012, capable of oligomerizing proteins engineered to contain the FK506 binding protein (FKBP), to trigger Fas via FKBP-linked receptor cytoplasmic domains in human keratinocytes. An FKBP chimera containing the Fas cytoplasmic domain targeted to the plasma membrane induced an up to 89% decrease in viability of keratinocytes, as reflected by the activity of constitutive promoters, in response to FK1012. Oligomerization of Fas, either with engineered Fas.FKBP by FK1012 or via antibody cross-linking of full-length Fas-induced cellular changes consistent with apoptosis. The lpr Fas point mutation abolished this effect. A Fas.FKBP construct unlinked to the membrane was fully active in this assay. Early developmental age or pre-treatment of cells with GM-CSF, TGF-beta, EGF, KGF, IFN-gamma, or phorbol ester failed to protect against Fas effects. These findings reveal that the Fas signal transduction pathway is active in keratinocytes, requires no induction, and dominantly overrides growth stimuli.     

Reversal of fusimotor reflex responses during locomotion in the decerebrate cat

The effect of brief trains of electrical stimulation, at 2, 3 and 20 x threshold (T), of cutaneous afferents in the medial plantar nerve on the discharges of single medial gastrocnemius static and dynamic gamma-efferents has been investigated at rest and during locomotion in a decerebrate cat preparation. The units were classified as dynamic (10 units) or static (10 units) indirectly on the basis of their resting and locomotor discharge characteristics. Responses were assessed by calculating the change in mean gamma-rate during the 100 ms after stimulus onset compared with a control period. At rest, most dynamic neurones were inhibited by stimulation at 2T (9 of 10 units) and above. In contrast, the resting responses of most static neurones were excitatory at 2T (9 of 10 units) and 3T, while 20T produced static gamma-effects that varied in sign. During locomotion the responses of both types of gamma-efferent were phase related. Two patterns were observed with dynamic units. For seven dynamic neurones, at stimulus levels of 2T (7 units) and above, responses during electromyogram (EMG) bursts were inhibitory while those between bursts were not significantly different from zero. However, for three other dynamic units, a phase-related reversal of reflex responses was observed at some stimulus intensities (always 2T, 3 units) comprising inhibition during, and excitation between, EMG bursts. For static neurones, inhibitory (never excitatory) responses occurred during walking at stimulus intensities of 2T (10 units) and above. The locomotor responses of static units were maximum during (3 units) or between (7 units) EMG bursts and were minimum in the opposite phase of EMG activity. A task-related reversal of reflex responses was thus generally apparent (9 of 10 units) to low intensity stimulation (2T) for static gamma-efferents during locomotion (inhibition) compared with rest (excitation). During locomotion there was a significant linear relation between the magnitude of response and the background gamma-rate for static units and those dynamic units that did not exhibit phase-related reflex reversal (total, 17 units). For dynamic gamma-efferents, inhibition at rest and during locomotion occurred at short (spinal) latencies which were not significantly different and are consistent with the involvement of the same interneuronal pathway. We conclude that pathways of opposite sign may dominate the responses of fusimotor neurones to low threshold cutaneous afferents from the plantar surface of the foot depending on behavioural context. Furthermore, the cutaneous reflex responses of both types of gamma-motoneurones during locomotion appear to vary with the source of the afferent input and do not constitute a general excitatory drive. The results are discussed in relation to the role and reflex control of the fusimotor system.     

Screening for amblyopia in preverbal children with photoscreening photographs. National Children's Eye Care Foundation Vision Screening Study Group

OBJECTIVE: This study aimed to determine the ability of healthcare professionals and lay volunteers to grade photoscreening photographs. DESIGN: The study design was a cross-sectional study. PARTICIPANTS AND INTERVENTION: One hundred children 3 years of age or younger received a complete ophthalmologic examination and were photographed using the Medical Technology Innovations (MTI) photoscreener. Twenty-six children had normal examination results, and the remaining 74 children had conditions that are of interest for pediatric screening, including strabismus, refractive error, media opacities, and ptosis. Eighteen volunteers, including pediatric ophthalmologists, pediatricians, ophthalmic technicians, health department nurses, Prevention of Blindness Society personnel, and Lions Club volunteers, graded each of the 100 photoscreening photographs. MAIN OUTCOME MEASURES: Sensitivity and specificity of vision screening and of photograph grading were measured. RESULTS: Results from various graders yielded sensitivities ranging from 37% to 88% and specificities ranging from 40% to 88%. No single grader achieved sensitivity and specificity both greater than 70%. The grading of the manufacturer's representative had a sensitivity of 43% and a specificity of 85%. Sensitivity decreased to 31% for strabismus and 18% for refractive error when the correct type of strabismus or refractive error was required to be considered true-positives. Results were not positively correlated with the ophthalmologic knowledge of the participant. CONCLUSIONS: The wide variability in sensitivities and specificities among graders indicates inconsistent photograph interpretation skills or deficient screening guidelines or both. For off-axis photoscreening as implemented by the MTI system to become a useful vision-screening method, additional photograph interpretation skill transfer may be beneficial, although not necessarily sufficient.