Human marrow erythropoiesis in culture. I. Characterization of methylcellulose colony assay

We examined the morphological and functional characteristics of human marrow erythrocytes cultured with a recently developed methylcellulose colony assay technique. Erythrocytic cells in various stages of development were observed, and a significant degree of maturational synchrony within individual colonies was noted. By light microscopy, colonies consisting of late normoblasts appeared compact, had an orange hue attributable to their hemoglobin, and demonstrated pseudoperoxidase activity, whereas colonies composed of early erythroblasts grew less compact or in clusters of smaller cell aggregates and showed no reddish tinge. Colonies possessing intermediate features were also observed. Maturational synchrony of individual colonies was confirmed using ransmission and scanning electron microscopy. The ultrastructure and cytochemistry of most immature cells were normal. The mature erythrocytes, however, were severely microcytic and hypochromic and contained one to several Heinz bodies. These defects in the cytoplasmic maturation of erythrocytes corresponded with impaired granulocytic maturation in culture, which we observed previously, and suggest environmental or nutritional defects in culture. Linearity of the method was confirmed using five normal bone marrows. Erythropoietin dose-responses observed in ten normal marrows were comparable to the previously reported results and revealed significant variation in individual plating efficiencies.     

Exogenous oxidants initiate hydrolysis of endothelial cell inositol phospholipids

Oxidants released from inflammatory cells contribute to the pathogenesis of acute inflammatory edema in many models. Chemically produced oxidants can reversibly alter the barrier properties of cultured endothelial and epithelial monolayers. This report examines the effects of nonlytic doses of H2O2 on endothelial cell lipids. H2O2 oxidized omega-6 fatty acids in the endothelial cells and initiated hydrolysis of endothelial cell phospholipids. When endothelial cells were exposed to peroxidized linoleic acid, it caused lysis of the cells at doses 1,000-fold lower than effective doses of H2O2. The phospholipid hydrolysis was directed primarily at the inositol phospholipids and consisted of both A and C type phospholipase activity. The phospholipase A hydrolysis resulted in increases in endothelial cell free fatty acids and lysophosphatidylinositol. The phospholipase C hydrolysis resulted in increases in diglycerides, phosphatidic acid, and inositol polyphosphate levels. The phospholipase C hydrolysis of phosphatidylinositol is known to activate protein kinase C in most cells. Stimulation of protein kinase C with phorbol- 12,13-dibutyrate increased albumin flux across endothelial monolayers and altered endothelial cell shape, similar to effects of oxidants. These data are consistent with the hypothesis that oxidant-initiated hydrolysis of endothelial cell inositol phospholipids contributes to oxidant-mediated reversible changes in endothelial monolayer barrier function.     

Liver transplant recipients: portable duplex US with correlative angiography

Twenty patients, aged 4 months to 58 years, were evaluated for liver transplantation by duplex sonography, and 15 transplantations were completed; 42 postoperative examinations were performed. Sonographic findings were correlated with seven preoperative and five postoperative angiographic evaluations. Preoperative duplex US findings included tumors, portal vein occlusion, varices, biliary obstruction, and variant vascular anatomy. Postoperative findings included hepatic artery occlusion, portal vein occlusions (one with cavernous transformation), portal vein stenosis, biliary obstruction, intrahepatic and extrahepatic fluid collections, and air in the portal vein due to ischemic bowel. Use of angiography allowed confirmation of the vascular abnormalities and demonstrated evidence of rejection in patients with normal Doppler waveforms. Duplex sonography is a valuable portable technique for evaluating these patients and can be used in triage of patients requiring angiography.     

Lipomatous tumors of the liver: evaluation with CT and US

Seven cases of lipomatous masses within the liver parenchyma were demonstrated with computed tomography (CT). Five of these cases were obtained from a retrospective review of 50 cases of renal angiomyolipoma in which the liver was adequately demonstrated. The other two cases were from the files of the Armed Forces Institute of Pathology and had no associated renal lesions. Three of the five cases were associated with tuberous sclerosis. In all seven cases, the fatty tumors appeared on CT scans as a well-defined, 0.8-13-cm mass, with attenuation coefficients of less than -30 HU. On ultrasound studies, the lesions were well circumscribed, highly echogenic, and similar to hemangiomas. While distinctly rare lesions, these lipomatous masses are not as unusual as the literature would indicate. One may anticipate such masses in patients with renal angiomyolipomas and in a relatively high percentage of those with tuberous sclerosis.     

Focal liver masses: differential diagnosis with pulsed Doppler US

Duplex Doppler ultrasound (US) was used in 68 consecutive patients with focal liver lesions, including 12 hepatocellular carcinomas, one cholangiocarcinoma, 37 metastases, 15 hemangiomas, one hemangioendothelioma, and two focal nodular hyperplasias. Of the hepatocellular carcinomas, six were diffusely hyperechoic, two were hypoechoic, two were single hyperechoic lesions, and two were multifocal and hyperechoic. All ten tumors with Doppler shifts of 5 kHz or above proved to be hepatocellular carcinomas. The other two hepatocellular carcinomas showed Doppler shifts of 3 kHz. In contrast, no hemangioma showed shifts above 0.7 kHz, and ten of the 15 gave no detectable signal. Of the metastases, 20 gave no signal and 17 had signals of up to 4 kHz. Three-kilohertz signals were also obtained from a cholangiocarcinoma, a hemangioendothelioma, and focal nodular hyperplasia. Correlation with angiographic findings suggested that the high-velocity Doppler signals were associated with large pressure gradients due to arteriovenous shunting. Duplex Doppler US can therefore aid in the differential diagnosis of diffuse and focal liver lesions.     

Trabecular bone mineral density measurement in vivo: use of the ratio of coherent to Compton-scattered photons in the calcaneus

Trabecular bone mineral density (TBMD) was measured in vivo in the calcaneus by a new method that uses the ratio of coherent to Compton-scattered photons arising from irradiation of a small volume of trabecular bone by a gamma ray source with highly collimated geometry. TBMD values for healthy men (22-77 years) were in the range of 180-357 mg/ml, and values for healthy women (18-73 years) were in the range of 160-321 mg/ml. In contrast, values in a small group of paraplegics were in the range of 90-199 mg/ml.