Glycogen in neuroblastomas. A light- and electron-microscopic study of 40 cases.

A light- and electron-microscopic review of 40 cases diagnosed at Children's Hospital of Pittsburgh as ganglioneuroblastoma, neuroblastoma, or small round cell tumor-probably neuroblastoma disclosed four cases that contained abundant glycogen. Two were unquestionable neuroblastomas by electron microscopy; one was primary in the adrenal gland, the other in the mediastinum. In the third case, a paraspinal tumor, the light-microscopic appearance was suggestive or neuroblastoma, but no catecholamine granules or neural processes were demonstrated in the material available for electron microscopy. The fourth case was an undifferentiated malignant tumor in the pectoralis muscle of a 12-year-old girl. By electron microscopy, neural processes were demonstrated and the tumor was classified as peripheral neuroblastoma. Of the remaining 36 cases, electron microscopy readily indicated a diagnosis of neuroblastoma or glangioneuroblastoma in 35 of them. In the other case, the tissue had been fixed in formalin and only a few catecholamine granules were found after an extensive search.

The effects of tissue processing on markers for T and B cells from solid tissues.

Suspensions of lymphoid cells from tissues have been used for the determination of the quantitative relationship between the T and B cell populations. The distribution of the lymphocytes within a given tissue, however, cannot be demonstrated once such a suspension has been prepared. Various methods of characterizing lymphocytes within tissues were evaluated. The method of tissue preparation can alter the capability of detecting the lymphocyte markers. Fluorescein-labeled anti-immunoglobulin sera reacted equally well with lymphocytes in tissue regardless of the method of tissue preparation. Complement-coated sheep erythrocytes were less effective in detecting lymphocyte markers in tissue sections than in cell suspensions. Quantitative assays of lymphocytes could be done in suspensions only. Unaltered sheep erythrocytes did not bind to T lymphocytes in tissue. T lymphocytes could be identified in tissue sections, however, by the use of anti-human T cell serum.

Conduction tissue anomalies in absence of the right superior caval vein.

The incidence of electrical instability of the heart is high in patients with absence of the right and persistence of the left superior caval vein when the latter connects to the coronary sinus. It has been suggested that a large coronary sinus may influence the susceptibility to arrhythmias. we studied the conduction tissues of 8 hearts from the cardiopathological collection of Children's Hospital of Pittsburgh. Six of these specimens had a persistent left superior caval vein connecting to the coronary sinus. There were 4 with absence of the right superior caval vein, 3 of which were in the group with persistent left veins. We evaluated the sinus node and the specialized atrioventricular junctional area, comparing them with known normals. The coronary sinus varied in size from through it. The size did not alter the histology of the adjacent conduction tissue. In contrast, the sinus node was abnormal in 3 of the 4 hearts with absent right superior caval vein. This may be the key factor in the development of arrhythmias when the right superior caval vein is absent or abnormal.

Fatal strychnine poisoning: application of gas chromatography and tandem mass spectrometry.

The history and toxicological findings in a case of suicidal fatal strychnine poisoning are presented along with a description of the analytical methods. Detection and quantitation of strychnine in body fluids and tissues was performed by gas chromatography (GC) with nitrogen-phosphorus detection, using organic extraction and calibration by a standard addition method. Strychnine concentrations in subclavian blood (1.82 mg/mL), inferior vena cava blood (3.32 mg/mL), urine (3.35 mg/mL), bile (11.4 mg/mL), liver (98.6 mg/kg), lung (12.3 mg/kg), spleen (11.8 mg/kg), brain (2.42 mg/kg), and skeletal muscle (2.32 mg/kg) were determined. Confirmation of strychnine in blood and tissue was performed by GC with detection by tandem ion-trap mass spectrometry (MS). GC-MS-MS analysis, employing electron ionization followed by unit mass resolution and collision-induced dissociation of strychnine, resulted in confirmatory ions with mass-to-charge ratios of 334 (parent ion), 319, 306, 277, 261, 246, 233, and 220. Additional confirmation was provided by GC-MS-MS-MS analysis of each confirmatory ion, revealing an ion fragmentation pathway consistent with the molecular structure of strychnine. The case demonstrates body tissue and fluid distribution of strychnine in a fatal poisoning and the application of tandem MS in medical examiner casework.

Increased microvascular permeability in canine endotoxic shock: protective effects of ibuprofen.

The ability of sodium ibuprofen to prevent endotoxin-induced changes in vascular permeability was examined in an anesthetized canine model. Ibuprofen was administered i.v. (3.75 mg/kg) in two pretreatment doses before the administration of Escherichia coli endotoxin (0.5 mg/kg). Serum and left thoracic duct lymph samples were collected for measurement of total protein and separation by polyacrylamide gel electrophoresis. Four protein fractions with molecular weights (MW) ranging from 60,000 to 1,000,000 were consistently analyzed. Administration of endotoxin alone resulted in hypotension and was accompanied by an increase in microvascular permeability as evidenced by increases in lymph flow rate, protein flux, lymph/plasma protein ratio (L/P), and permeability-surface area product (PS). Pretreatment with ibuprofen attenuated the increase in permeability as reflected by significantly lower lymph flow rate, protein flux, L/P, and PS. Electrophoretic data illustrate partial to complete protection for all four MW fractions. These results suggest that endotoxin damages microvascular integrity and increases extravasation of macromolecules as great as 1,000,000 MW. This damage is attenuated significantly by pretreatment with ibuprofen.

Effects of methylprednisolone upon vascular permeability changes in endotoxic shock.

This study was designed to examine glucocorticoid effects on increases in vascular permeability caused by endotoxic shock in dogs anesthetized with pentobarbital sodium (30 mg/kg). Methylprednisolone sodium succinate was administered IV in two doses (30 mg/kg each) before Escherichia coli endotoxin was administered (0.5 mg/kg). Samples of serum and lymph from the left thoracic duct were collected for measurement of total protein and separation by polyacrylamide gel electrophoresis. The same four protein electrophoretic fractions with molecular weights (M.W.) ranging from 60,000 to 1,000,000 were consistently chosen for analysis. Methylprednisolone treatment given prior to endotoxin administration resulted in an attenuation of the early increase in total protein flux and lymph to plasma protein (L/P) ratio and prevented significant increases in the permeability surface area product observed in the group given endotoxin alone. Endotoxin administration alone resulted in significant increases in the permeability-surface area product and L/P ratio for all four electrophoretic fractions. Pretreatment with glucocorticoid partially attenuated the increase in the L/P ratio for only those fractions of 100,000 M.W. or less. These results suggest MP provides partial but not complete protection from increases in vascular permeability during endotoxic shock.

Osteosarcoma arising in osteogenesis imperfecta.

We report a case of osteosarcoma arising in osteogenesis imperfecta. The characteristic radiographic and pathological findings are discussed and emphasized to warrant a biopsy.

Blood volume changes in endotoxin shock.

Injection of E. coli endotoxin in dogs promotes hemodynamic changes in circulating blood volume and blood components. Adult mongrel dogs were given endotoxin and studied for 3 hours. Red-blood-cell (RBC) volume and whole-blood volume was measured using the Cr-51 RBC labeling method. Hematocrit and RBC counts were taken at various time periods. Whole-blood volume was not found to change significantly over the test period; however, plasma volume demonstrated a significant decrease by 60 minutes with a parallel increase in RBC volume. Hematocrit increased significantly by 60 minutes and continued its rise to a 30% increase at 3 hours. Circulating RBC counts demonstrated a 26% increase. RBC precursors such as reticulocytes and normoblasts were counted using standard microscopic methods and were observed to increase after injection of endotoxin. These data suggest that the increases in hemoconcentration after administration of endotoxin are caused not only by the loss of plasma from the circulation but also by release of RBCs from blood-forming organs.