Clinical heterogeneity of adhalin deficiency

STUDY DESIGN: An assessment was made of the efficacy of a combined laminoplasty and foraminotomy operation for patients with coexisting myelopathy and unilateral radiculopathy. The procedure was done in 17 patients. OBJECTIVES: The patients were followed with lateral flexion and extension radiographs, computed tomography scans, and an assessment system specially designed to qualitatively evaluate the patients' neurologic status. Follow-up period averaged 4 years (range, 2.1-9.3 years). SUMMARY OF BACKGROUND DATA: Excellent-to-good results were obtained for 76% (13 of 17) of the patients without any significant functional compromise based on the radiographs. Sixteen nerve roots were decompressed with a less than 25% foraminotomy, whereas eight were decompressed by a 25%-50% foraminotomy without serious neurologic damage, except for one patient. The neurologic results appeared unrelated to the extent of foraminotomy. METHODS: A refined procedure for combined laminoplasty and foraminotomy was reviewed retrospectively in terms of neurologic outcome and radiographic data. RESULTS: The present series is small, and results are not comparable directly with other methods. The procedure appears effective for myelopathy and radiculopathy. This procedure is applicable to patients with myelopathy and coexisting nerve root impingement anterolaterally or in the neural foramen. CONCLUSION: The combined laminoplasty and foraminotomy operation may provide greater neurologic improvement in patients with coexisting myelopathy and unilateral radiculopathy, while maintaining cervical spine stability after surgery.     

Ultrastructural changes in skeletal muscle of selenium-vitamin E-deficient chicks

Chicks fed a semisynthetic basal diet deficient in selenium and vitamin E for 14 to 22 days developed skeletal myodegeneration and exudative diathesis. Chicks fed the basal diet supplemented with either 0.2 ppm of selenium (as selenite) or 100 IU alpha-tocopherol acetate/kg were protected from deficiency disease, but chicks fed the basal diet plus 0.4% L-cystine were not protected. Pectoral muscles of deficient chicks were red and edematous. Light and electron microscopic study of affected muscles revealed fibers with hyaline and granular degeneration. In hyalinized fibers, the initial ultrastructural alterations were increased density of the sarcoplasm and myofibrils, dilatation of sarcoplasmic reticulum, formation of subsarcolemmal vacuoles, and disruption of mitochondrial membranes. In later stages, alterations in these fibers included myofibrillar disruption and lysis, nuclear pyknosis and lysis, disruption of the plasma membrane with persistence of basal lamina and scattered adhering satellite cells, and eventual invasion by macrophages. In fibers with granular degeneration, the ultrastructural observations included decreased density of the sarcoplasm, prominent mitochondrial swelling and distortion, and multiple foci of myofibrillar lysis that eventually coalesced to produce generalized lysis. Prominent vascular lesions associated with exudative diathesis were present in degenerated muscle but were not considered to precede development of fiber alterations. Affected blood vessels had endothelial cells with mitochondrial damage and accumulations of cytoplasmic dense bodies and areas of endothelial disruption with adhering fibrin thrombi.     

Ultrastructural localization of microliths in salivary glands of cat

Although microliths occur in normal human salivary glands and may be an aetiological factor of sialadenitis, little is known of their natural history. In an attempt to remedy this, we investigated a large archival collection of normal and experimental feline parotid, submandibular and sublingual salivary glands. In submandibular and sublingual glands, microliths were detected ultrastructurally in: all types of acinar secretory cells; myoepithelial cells; ductal cells; lumina; intercellular spaces; basement membrane; stroma; macrophages; multinuclear giant cells; and neutrophils. Microliths were not detected ultrastructurally in parotid glands. Microliths appear to form in acinar cells during autophagy and in stagnant secretory material in lumina. Microliths appear to be removed by secretion in the saliva, discharge from cells laterally and basally, and engulfment by macrophages. There appears to be a turnover of microliths, which possibly is upset by secretory inactivity with a resulting accumulation that leads to localized obstruction and sialadenitis.     

Ultrastructural osteopontin localization in papillary stones induced in rats

OBJECTIVES: To detect in situ the precise osteopontin (OPN) localization in papillary stones. METHODS: Immunocytochemical labelling procedures are applied to detect OPN localizations in crystalline material of renal papillary stones. The tissue-processing procedure for electron microscopy, which includes OsO4 postfixation, preserves both immunocytochemical OPN reactivity and cellular membrane contrast up to the ultrathin section. Reflection-contrast light microscopical images are correlated with high resolution transmission-electron microscopical observations from consecutive ultrathin epon sections. RESULTS: Preserved crystalline material in interstitial and peripheral papillary stones is recognized as calcium oxalate monohydrate. After section incubation with markers conjugated to an antibody against OPN (alpha OPN) the crystals are converted into ghosts. In the ghosts, alpha OPN markers are present around microcrystals. The size of these microcrystals ranges from several nanometers to micrometers. It is observed (due to the OsO4-preserved membranes) that interstitial cells are separated from the stone surfaces by unidentified extracellular material, also present in the center as a stone matrix. CONCLUSION: The microcrystal-growth inhibitor OPN is detected in situ in interstitial stones induced in the rat's papilla and at the surface of the papilla.     

Ultrastructural localization of concanavalin A in the perfused rat heart

The purpose of this investigation was to assess the alteration in serum free fatty acid concentrations during heat stress and dehydration. Each subject was exposed to heat stress in an environment chamber on 2 separate occasions. During the first exposure the subjects remained seated until the core temperature was elevated 1.4degreesC resulting in a mean weight loss of 1.66 kg due to dehydration. The second condition involved water replacement equal to the weight loss of the initial dehydration condition. Blood samples were obtained prior to heat exposure, when the core temperature was elevated 0.7degreesC and 1.4degreesC. They were subsequently analyzed for free fatty acids (FFA), glucose and lactin acid. Heart rates and core temperatures were monitored at 4 min intervals. During the dehydration condition the mean change in serum FFA was 0.9 muEq/ml in contrast to 0.2 muEq/ml for the rehydration condition. Serum levels of glucose increased moderately throughout the exposure (8 mg-%).     

Ultrastructural localization of DNA in immune deposits of human lupus nephritis

DNA molecules were revealed in the glomerular wall of lupus nephritis patients by applying two specific colloidal gold cytochemical approaches at the electron microscope level: immunocytochemistry using a monoclonal anti-DNA antibody in conjunction with protein A-gold and enzyme-gold cytochemistry using DNAse-gold complexes. Application of both techniques has demonstrated that DNA molecules are preferentially located over the electron-dense deposits found in the glomerular basement membrane and mesangial matrix of SLE patients, as well as over the nuclei. Their distribution within the glomerular wall was correlated with electron-dense immune deposits revealed by anti-light chain antibodies. In normal control kidney, DNA labeling was restricted to the cell nuclei. Several control experiments have demonstrated the high specificity of the results. These data thus suggest a possible role for DNA as an antigenic component in the formation of immune complexes.     

Immunocytochemical localization of desmin in human fetal skeletal muscle

INTRODUCTION: A comparative evaluation of four air samplers was performed using bioaerosol collection in the outdoor environment. METHODS: Test samplers used included a Rotorod, a Kramer-Collins suction trap, an all-glass impinger (AGI-30), and a high-volume cyclonic liquid impinger (SpinCon). All samples were analyzed microscopically for spores and pollen. The two collectors providing a liquid sample (AGI-30 and SpinCon) also were analyzed for specific allergen content by enzyme-linked immunoassay. RESULTS: The SpinCon collected a larger number of spores than the other devices. The number of spores collected by this unit per volume of air sampled was comparable to the AGI-30. The Rotorod and Kramer-Collins collected a lower number of spores per unit of air but collected a larger number of pollen grains per volume sampled. Alternaria allergens Alt a I and GP70 were collected by both liquid impingers; however, the SpinCon collected more Alt a I and the AGI-30 collected more GP70. CONCLUSIONS: The SpinCon is a device that is capable of efficiently sampling a high volume of air and concentrating it in a form that can be analyzed for the presence of spores and fungal allergens. It is less useful for collecting intact pollen grains. Pollen allergen quantitation has not yet been performed on the SpinCon effluent.     

Ultrastructural localization of DNA in leukemic cells using osmium ammine B

In order to determine whether there were any differences in distribution of nuclear DNA between acute lymphocytic leukemia (ALL) and acute myelogenous leukemia (AML), the localization of DNA in blasts from the bone marrow or buffy coat of 30 patients with ALL and 30 patients with AML was examined ultrastructurally by staining with osmium ammine B. By the ultrastructural cytochemistry, DNA in ALL cells was clumped in the nuclei, while in AML cells, it was dispersed. DNA had accumulated around the nucleoli of some blasts, and flecks of DNA were observed in nucleoli of a majority of blasts. The perinucleolar and intranucleolar DNA distribution could be classified into four types. The types with abundant perinucleolar DNA were frequently observed in ALL blasts, while the majority of AML blasts showed scant perinucleolar DNA. The types with intranucleolar flecks of DNA were more prominent in leukemic cells than in normal immature leukocytes. In conclusion, the pattern of distribution of DNA in the nuclei of leukemic cells differs between ALL and AML.     

Ultrastructural localization of filamentous actin within neuronal interphase nuclei in situ

Previous biochemical studies utilizing isolated nuclei and nuclear matrices have shown actin to be a constituent of the interphase nucleus. In addition, recent ultrastructural work has shown the presence of actin and myosin within nuclei of interphase cells in situ. It was unclear, however, whether this intranuclear actin is present in the unpolymerized globular actin or the filamentous (F)-actin form. The present work, using confocal microscopy and ultrastructural cytochemical techniques, demonstrates the presence of F-actin within interphase nuclei of intact, uncompromised, dorsal root ganglion neurons in vitro and in vivo. Labeling by FITC-phalloidin detected the presence of intranuclear F-actin adjacent to the nucleolar periphery in a small fraction of cells in vitro, an observation confirmed by three-dimensional reconstruction. Ultrastructural analyses of cells exposed to heavy meromyosin (HMM), showed the presence of typical "arrowhead" complexes. The observation that these complexes were associated with nucleoli confirms that the intranuclear ligand detected by FITC-phalloidin indeed represents F-actin. Postembedding labeling with HMM conjugated to 20-nm colloidal gold (HMM-Au20) resulted in labeling similar to that obtained with HMM. However, HMM-Au20 was found to label a much larger fraction of cells, both in vitro and in vivo, than did FITC-phalloidin or HMM. This finding indicates that labeling with HMM-Au20 more accurately reflects the extent of actin polymerization in nuclei. Results from double labeling with HMM-Au20 and an antibody to alpha-sarcomeric actin confirmed that only a small amount of nuclear actin is in the F-form. Together, these results represent a first ultrastructural demonstration of the presence of F-actin in nuclei of neurons. While the role of nuclear F-actin has yet to be defined, the results suggest that F-actin may represent a component of the molecular motor responsible for the dynamic positioning of specific chromatin domains into the tissue-specific, nonrandom patterns observed in many cell types.     

Ultrastructural localization of P2X3 receptors in rat sensory neurons

We used isolated IgG antibodies selective for P2X3 receptors to study the ultrastructural distribution of these receptors in rat sensory neurons. In trigeminal ganglia, P2X3 receptor immunoreactivity occurred in small and large nerve cell bodies and their processes. Endoplasmic reticulum and Golgi apparatus were heavily stained; cytoplasmic matrix was faintly to moderately stained. In synaptic glomeruli in lamina II of cervical dorsal horn, P2X3 receptor-immunoreactive core terminals were postsynaptic to unlabelled vesicle-containing dendrites and axons. In the nucleus of the solitary tract, receptor-positive boutons synapsed on dendrites and cell bodies and had complex synaptic relationships with other axon terminals and vesiculated dendrites. These observations identify sites from which ATP could be released to influence sensory signalling within the central nervous system.     

Ultrastructural localization of nitric oxide synthase immunoreactivity in the cat ventrobasal complex

This study describes the ultrastructural localization of nitric oxide synthase (NOS) immunoreactivity in the cat ventrobasal complex. NOS immunoreactivity was found in the cell bodies and dendrites of local circuit neurons and in vesicle-containing profiles. The vesicle-containing profiles could be divided into two classes, those of dendritic origin (presynaptic dendrite boutons) and those of axonal origin. The NOS labelled axon terminals varied in size and packing density and were principally located in the extra-glomerular neuropil. These boutons presented a range of morphologies and it was not possible to determine the probable source based on morphological criteria. The NOS immunoreactive presynaptic dendrite boutons were found both within and outside glomeruli and established both pre- and post-synaptic relationships with other elements. Post-embedding GABA immunocytochemistry showed that some NOS immunoreactive axonal boutons and presynaptic dendrites were also immunopositive for GABA. This finding suggests that some of the NOS labelled axonal boutons are of local circuit neuron origin. These results suggest that local circuit neurons in the cat ventrobasal complex might be involved in specific, short range interactions using GABA and longer, more global interactions using nitric oxide.     

Ultrastructural localization of actin in the cell body of rat spinal ganglion neurons

We used phalloidin staining and immunocytochemistry at the light and electron microscope level to determine the localization of actin in the cell bodies of rat spinal ganglion neurons. The results show that actin is mostly concentrated along the periphery of the neuronal perikaryon, including the perikaryal projections. This localization places actin in a strategic position to be influenced by incoming signals and to produce mechanical tensions able to shape the perikaryal surface.     

Ultrastructural localization of thyrocyte acid phosphatase in the presence of thyrocyte hyperfunction in rats

Acid phosphatase was found by electron microscopy in the lysosomes which appeared in great numbers in the follicular cells of rat hyperplastic thyroid gland. The other types of granules (mature secretory granules and lipids) whose amounts were also greatly increased in cases of functional thyrocyte strain were also nonreactive. The lysosomes were subdivided into three main groups according to distribution of the reaction product: the lysosomes with dense homogeneous deposit and with deposit in the form of densely or loosely packed dark round granules. The lysosome heterogeneity was apparently connected with their different functions also found within the colloid droplets in the form of inclusions of rarely located dark granules. The authors believe such granules to be the result of the merging of the colloid droplets and lysosomes. The acid phosphatase of the latter participated in the hydrolysis of the product of cell secretion with the formation of active substances.     

Ultrastructural localization of nerve terminals containing nitric oxide synthase in rat adrenal gland

The effect of hyperosmolality (300, 320 mosmol/kg H2O) and angiotensin II (A II, 10(-8) and 10(-6) M) was tested on a total of 64 neurons within the periventricular part of the anteroventral third ventricle (AV3V) region in brain slice preparations obtained from ovariectomized (OVX) rats with or without chronic treatment of estradiol-17 beta (E2). Hypertonic perfusion with a 320 mosmol/kg H2O but not a 300 mosmol/kg H2O medium caused a significant increase in the firing discharge rate in OVX animals. Perfusion with either hypertonic medium had no effect in E2-treated rats. The neuronal firing discharge rate of neurons in OVX rats was increased following perfusion with 10(-6) M A II, but not affected following perfusion with 10(-8) M A II. In E2-treated rats, perfusion with neither 10(-8) nor 10(-6) M A II had any effect. These data suggest a dynamic relationship between the ovarian endocrine function and the central mechanisms regulating dipsogenic behavior and/or release of vasopressin in the female rat.     

Ultrastructural localization of GLUT 1 and GLUT 3 glucose transporters in rat brain

Owing to the complexity of the parathyroid hormone's metabolic interactions, clinical hypoparathyroidism is one of the most difficult of all endocrine disorders to treat. Therefore, causative treatment of this disorder by transplantation of parathyroid glands is highly desirable. We have recently documented the long-term in vivo function of iso- and allotransplanted rat parathyroid tissue without systemic immunosuppression in an animal model. In view of the potential clinical use of this method, human parathyroid tissue has been microencapsulated and transplanted over the highest immunological barrier. In a controlled, long-term animal study in the parathyroidectomized rat, the effect of microencapsulation on xenotransplanted human parathyroid tissue was evaluated over 30 weeks (native and microencapsulated parathyroid tissue = 40 rats respectively). Functionally, human parathyroid tissue was able to replace that of the rat. All animals that had received microencapsulated parathyroid tissue were normocalcemic for 16 weeks; 27/40 at the end of the study. In contrast, serum calcium concentrations dropped to post-parathyroidectomy levels within 4 weeks in those animals that had received native tissue only. Histologic evaluation of the explanted, functionally successful xenografts showed vital parathyroid tissue inside intact microcapsules surrounded by a small rim of fibroblasts. Avital fibrotic remnants were demonstrated in animals with non-encapsulated parathyroid tissue. Thus, we have established the feasibility of microencapsulation of human parathyroid tissue, preserving its viability over long periods in vivo even if xenotransplanted. In combination with an improved tissue culture method, transplantation of human parathyroid tissue and maintenance of its physiological function is reproducibly achieved without postoperative systemic immunosuppression over the highest transplantation barrier. This may be a crucial step towards the first clinical application of this method.     

Quantitative histochemical evaluation of normal human skeletal muscle

Skeletal muscle tissue was obtained by open biopsy from the vastus lateralis and peroneus brevis muscles from 12 and 16 healthy paid volunteers, respectively. Frozen sections were examined with standard histochemical methods. Central nuclei, small and large angular fibers, and small round fibers were the most common "abnormalities" present. The number of fibers of these types were quantified, along with other more rare deviations from normal morphology. Several of the abnormalities were more common in the peroneus brevis than in the vastus lateralis.