Liposarcoma. An ultrastructural study of 15 cases.

Fifteen liposarcomas from 13 patients were examined by electron microscopy. These included nine primary tumors, four recurrent tumors after primary surgery or irradiation, and two metastatic lesions. Twelve of the liposarcomas were located in the thigh, and 11 were of the myxoid variety. All neoplasms were composed of cells having the ultrastructural characteristics of some stage of lipoblastic differentiation, i.e., lipid droplets, micropinocytotic vesicles, glycogen, external lamina, intermediate filaments, Golgi apparatuses, rough and smooth endoplasmic reticulum, and mitochondria. The nuclear-cytoplasmic ratio and nuclear pleomorphism were related inversely to the size and number of lipid droplets. Lipoblasts were frequently in close association with capillaries and pericytes, and in four cases lipid droplets were found in pericytes. Multivacuolated, mitochondria-rich lipoblasts, resembling brown fat cells, also were seen. Most tumors contained lipid-free, poorly differentiated mesenchymal cells that showed a continuum of morphologic differentiation to cells that closely resembled early lipoblasts that contained nonmembrane-bound lipid vacuoles. Fibrolipoblasts, cells containing lipid droplets and abundant rough endoplasmic reticulum, were observed only in well-differentiated liposarcomas. Some soft tissue sarcomas contain vacuolated cells that simulate lipoblasts by light microscopy but prove to be reactive or malignant fibroblasts, histiocytes, or smooth muscle cells ultrastructurally. Therefore, use of electron microscopy may be necessary to establish the line of differentiation in these neoplasms.

Lafora-body disease with optic atrophy, macular degeneration and cardiac failure.

A 17-year-old boy was admitted to hospital in acute cardiac failure and psychosis. The clinical course, EEG records and tissue diagnosis, including biopsies of brain, skin, skeletal muscle, peripheral nerve and liver were compatible with Lafora-body disease (LBD). Unusual features were those of optic atrophy and macular degeneration, signs generally regarded as negative criteria for the diagnosis of this disease. We also present the findings on endomyocardial biopsy which was performed because cardiac failure as an early symptom of LBD has not been previously described. The patient died in status epilepticus a few months after discharge from hospital.

Biochemical and morphologic changes in rabbit lung following endotracheal instillation of zymosan particles.

In order to establish a model of lung disease in which the usefulness of potential antiinflammatory compounds can be evaluated, we have analyzed the biochemical and cellular responses of rabbits to zymosan deposition in their lungs. A suspension of zymosan particles was instilled into the lungs of rabbits using an intratracheal catheter. Because the influx of leukocytes and the transudation of plasma into affected lungs was expected to contribute to the total cellular enzyme and protein levels, lungs were homogenized and assayed after various time intervals for six cellular enzymes and for protein content. After one day, alkaline phosphatase and neutral protease levels were elevated by 90% and 50%, respectively, above normal values. After two and three days, all of the pulmonary enzymes assayed displayed maximal two- to fourfold increases in their levels of activity. After seven days, only the alkaline phosphatase and neutral protease levels remained slightly elevated by 50% and 30%, respectively. Histologic analysis revealed focal and diffuse intraalveolar, interstitial, peribronchiolar, and perivascular accumulations of macrophages, granulocytes, and lymphocytes. Severe pulmonary edema, evident microscopically after one to three days, correlated well with 100% increases in both the wet weight and protein content of the lungs. In control experiments, the intratracheal infusion of saline solution minus zymosan particles resulted in a variety of enzymatic changes in the lungs after three days, which could be distinguished both enzymatically and histologically from those following zymosan deposition; histopathologic analysis revealed a pattern of intravascular congestion with erythrocytes, edematous thickening of alveolar septa, and focal intraalveolar hemorrhages, but with no inflammatory infiltration. In summary, this study demonstrates the time course of an experimental model for acute and chronic lung inflammation, the extent of which may be quantitatively evaluated using cellular enzymatic markers.

A biomechanical evaluation of suture anchors in repair of the rotator cuff.

Repair of the rotator cuff requires secure reattachment, but large chronic defects cause osteoporosis of the greater tuberosity which may then have insufficient strength to allow proper fixation of the tendon. Recently, suture anchors have been introduced, but have not been fully evaluated. We have investigated the strength of suture-to-anchor attachment, and the use of suture anchors in repairs of the rotator cuff either to the greater tuberosity or the lateral cortex of the humerus. The second method gave a significant increase in the strength of the repair (p = 0.014). The repairs were loaded cyclically and failed at low loads by cutting into bone and tendon, casting doubt on the integrity of the repair in early mobilisation after surgery. Repairs with suture anchors did not perform better than those with conventional transosseous attachment.

Allografted keratinocytes used to accelerate the treatment of burn wounds are replaced by recipient cells.

Cultured keratinocytes were used as allografts on burn wounds in two patients. In both patients successful covering of the wounds was obtained. DNA fingerprinting of the epidermis covering the wounds 21 days later showed that the cultured keratinocytes were replaced by the patients' cells.

Experimental reclamping of free-flap pedicles: the effect of prolonged stasis on the anastomoses and clamp sites.

The effect of static blood in direct contact with areas of microvascular anastomoses and previous clamp application for prolonged periods of time has been investigated. The free groin flap was used as a model in 27 white rabbits. The flap pedicle vessels were reclamped proximal to the anastomoses and areas of previous clamp application for periods of time varying between 30 minutes and 4 hours after 15 minutes of blood flow over these areas. A 100 percent patency rate was achieved despite the long periods of reclamping. Histologically, minor intimal damage was visible in the immediate period following anastomoses and clamping of the vessels. After 2 weeks, despite a thickened myofibroblastic intimal lesion, an intact endothelial layer was observed. No evidence of thrombosis could be demonstrated in either period. We postulate that vessels carefully treated and with technically well-performed anastomoses can be regarded as "normal" vessels after 15 minutes of blood flow over the anastomoses and clamp sites. We suggest that when required, microvascular clamps may then be reapplied without risk for prolonged periods of time despite static blood being in contact with these areas.

Semiquantitative postembedding characterization of intermediate filaments in central nervous system lesions using immunoelectron microscopy.

Standardized postembedding immunoelectron microscopy was performed to demonstrate glial fibrillary acidic protein (GFAP) and vimentin in individual intermediate filaments to determine the diagnostic value of demonstrating ultrastructural and immunophenotypic characteristics of intermediate filaments in routine brain biopsy specimens. Dual expression of GFAP and vimentin was observed in the astroblastoma and astrocytes of Alexander's disease. The antigen availability for vimentin, however, was too low to allow reliable assessment of the GFAP:vimentin ratio in individual intermediate filaments and/or filament bundles. In meningioma, only vimentin positive intermediate filaments were found. GFAP positive intermediate filaments were present in all other specimens except the oligodendroglial components of the mixed glioma, which were devoid of intermediate filaments. GFAP positivity in the filamentous periphery and electron-dense core of Rosenthal fibers was demonstrated. Technical and tissue processing factors had a significant effect on particle density values obtained for individual specimens. Although the number, distribution, and density of glial intermediate filaments varies in different astroglial entities, correlation of particle density values determined by immunoelectron microscopy with relative GFAP concentrations in different lesions requires utmost caution. Nevertheless, application of the postembedding approach to routinely fixed biopsy specimens indicated an association of different entities with the exclusive presence of GFAP and/or vimentin in individual intermediate filaments, thus emphasizing the diagnostic value of intermediate filament typing for pathological characterization.

Spontaneous rupture of adult Wilms' tumor.

Wilms' tumor is uncommon in adults, and spontaneous retroperitoneal hemorrhage as the presenting sign of renal malignancy also is rare. A 22-year-old man with spontaneous rupture of a Wilms' tumor, who died within 5 months, is described. The factors that probably contributed to his rapid demise included preoperative tumor spilling, unfavorable histology, and the omission of radiotherapy owing to poor patient compliance. Although Wilms' tumor is known to have a poorer prognosis in adults than in children, the reason for this is not known. Due to the limited number of cases reported, the optimal treatment for adult Wilms' tumor remains to be defined. However, the recent literature indicates that radical surgery combined with maximal radiotherapy dosage and modification of the chemotherapy schedules used in children may be required to improve the prognosis for adults with Wilms' tumor.

The influence of paraquat on the in vitro oxygen consumption of rabbit lung.

The effects of paraquat on the aerobic metabolism of rabbit lung slices were investigated. The oxygen consumption of lung slices was examined at different oxygen tensions and in the presence or absence of glucose as substrate in a Krebs-Ringer phosphate medium. In an air phase, the oxygen consumption of control lung tissue with glucose in the medium was practically the same as the oxygen consumption without glucose over a 3-hour period. In a 100% oxygen phase, glucose induced a marked increase in oxygen uptake, which persisted for about 2 hours. Thereafter, a decrease occurred in oxygen consumption which was notably faster than that of lung tissue without added glucose. With 10 mM glucose in the medium, paraquat (10(-5)M and 10(-3)M) immediately stimulated the oxygen consumption of lung slices. Although the initial increase in aerobic metabolism seemed to be glucose-dependent, the secondary inhibitory effect of paraquat appeared to be of the same magnitude in the presence or absence of glucose. Both the initial stimulation as well as the secondary inhibition were much more pronounced in a 100% oxygen atmosphere than in an air phase. These results indicate that the rabbit lung is sensitive to paraquat toxicity in vitro, and confirm that oxygen and paraquat supplement each other's toxicity in the lung.

The effect of paraquat on the respiration of lung cell fractions.

The in vitro effects of paraquat on the aerobic metabolism of lung and liver homogenates, as well as on the oxygen consumption of isolated rat and rabbit lung mitochondria, were investigated. It was found that the endogenous oxygen uptake of a 40% lung homogenate was similar to that of a 10% liver homogenate, and that succinate (20 mM) was well oxidized by both homogenates. About 14% of the basal respiration rate was due to cyanide-insensitive oxidative systems in lung and liver homogenates. Paraquat (1 mM and higher concentrations) induced an acute and highly significant increase in the cyanide-insensitive oxygen utilization. We were able to recover about 5% of the protein in the supernatant (1,000 g) of the original homogenate as mitochondrial protein. Only results obtained from mitochondrial preparations with a respiratory control ratio between 2,7 and 3,3 were used in the final analyses, and the normal state 3 respiration values of rabbit and rat lung mitochondria were 45,2 +/- 5,3 and 38,7 +/- 2,9 nmol oxygen per mg protein per minute respectively. Paraquat in concentrations up to 0.5 mM had no significant effect, but higher concentrations up to 1 mM induced a highly significant inhibition of mitochondrial oxygen consumption. When slices were pre-incubated with 1 mM and with 0,01 mM paraquat in the incubation medium, the oxygen consumption of the mitochondria isolated after 2 hours and 4 hours respectively, was significantly reduced.

The effect of oxygen and paraquat on the 14C-glucose oxidation of rabbit alveolar macrophages and lung slices.

In this study, we measured the effects of different concentrations of paraquat (0,01 mM and 1,0 mM) on the 1-14CO2 and 6-14CO2 production of rabbit lung slices and isolated alveolar macrophages, in 20% and 95% oxygen phases respectively. A 95% oxygen phase induced an increase in the 6-14C-glucose oxidation of contorol lung slices over a 3-hour period, while the increased activity of the pentose pathway over the first 2 hours started to decline during the third hour of incubation. Paraquat (1,0 mM) in 20% oxygen caused a consistent increase in the 6-14CO2 production by lung slices, but in a 95% oxygen phase gradually inhibited the 6-14C-glucose oxidation over a period of 3 hours. The pentose phosphate pathway was highly significantly stimulated by 1,0 mM paraquat in 20% and 95% oxygen over 3 hours. When isolated alveolar macrophages (viability 95%) were incubated in a 20% and a 95% oxygen phase respectively, both the 6-14C-glucose and 1-14C-glucose oxidation rates were significantly inhibited by 1,0 mM paraquat after 1 hour. Our results confirmed the initial increase in glycolytic metabolism induced by paraquat, but also inhibited by paraquat when lung slices were incubated in a 95% oxygen phase. The fact that the glucose metabolism in alveolar macrophages is more sensitive to paraquat exposure than that of cells in lung slices may be related to the genesis of the intra-aveolar pulmonary lesions described in the literature.

The effect of paraquat on the aerobic metabolism of rabbit alveolar macrophages and lung fibroblasts.

In this study the effects of paraquat on the aerobic metabolism and viability of isolated rabbit alveolar macrophages and lung fibroblasts were investigated, and compared with the effects of other known metabolic inhibitors, i.e. sodium fluoride (NaF) and potassium cyanide (KCN). The manometrically and polarographically determined endogenous oxygen consumption of lavaged alveolar macrophages compared very well (180,9 +/- 35,8 and 169,3 +/- 26,8 nmol per 10(6) viable cells per hour respectively). Exogenous glucose (10 mM) and autologous serum (1:3 v/v) added to the medium had no significant effect on the basal respiration rate. The mean cell protein content, determined by the micro-Kjeldahl and Lowry techniques, amounted to 242,6 +/- 37,6 microgram/10(6) macrophages. Paraquat (2 mM), like NaF (20mM) and KCN (5 mM), decreased the viability of the macrophages far less than it did the oxygen utilization of the viable cells, and resulted in an 80% inhibition of oxygen uptake. In contrast, paraquat (1 mM) induced a marked stimulation (230%) of the cyanide-insensitive respiration of alveolar macrophages. The concentrations of paraquat (nmol/10(3) cells) which reduce macrophage metabolism to almost zero were virtually non-toxic to fibroblasts, as measured by their oxygen consumption.

Ultrastructure of the capsule of the rabbit adrenal gland.

The capsule of the normal rabbit adrenal gland was examined with the electron microscope, and three layers are described. The outermost layer consists of collagen and elastic fibrillae, with cytoplasmic processes of fibroblasts in between. Myofibroblasts with parallel cytoplasmic filaments, dense bodies at the inner surface of the plasma membrane and an irregular nuclear outline were seen in the middle layer. The presence of unmyelinated nerves and an axon terminal near to the myofibroblasts of the middle layer raised the possibility of innervation of these cells which could have a contractile function. The basal lamina of the fenestrated capilaries in the inner vascular layer is occasionally fused with that of the outer zona glomerulosa, suggesting a probable route for blood supply and secretion.

Digital papular calcific elastosis: a histopathological, histochemical and ultrastructural study of 20 patients.

In 1960, 5 patients were reported with a condition termed "degenerative collagenous plaques of the hands". Ultraviolet light and chronic pressure seem to be the inciting factors. Clinically, it is characterized by keratotic, translucent papules, in linear array, on the radial border of the hands. Histology shows hyperkeratosis and dermal elastosis. The current study documented the histopathological, histochemical and ultrastructural features of 20 patients with this condition and considered the differential diagnoses. The findings were in accordance with those of previous studies, but peculiar, basophilic elastotic masses (BEMs), were a prominent finding. Ultrastructurally, elastic tissue appeared in the form of degenerated non-fragmented and fragmented elastic fibres, degenerated fibres showing foci of increased density, with or without a fibrillar component and as amorphous, structureless material. BEMs arose almost exclusively from degenerated elastic tissue and contained calcium. Disintegrated collagen may be incorporated in the amorphous material and, subsequently, form a minor part of BEMs. Solar elastosis, and its variants, show some similarity to degenerative collagenous plaques of the hands, but lack the constant presence of these calcified angular elastotic masses. BEMs show resemblance to elastic globes, and elastotic bodies found in elastotic nodules of the anthelix and cartilage-hair hypoplasia. In the clinical setting of "degenerative collagenous plaques of the hands", BEMs seem to be unique and due to their extensive presence serve as a non-specific marker for this condition. Digital papular calcific elastosis therefore seems to be more appropriate nomenclature.

The adrenal cortex in hypercholesterolaemic rabbits: histochemical and electron microscopical changes.

The adrenals of rabbits on a cholesterol-rich diet for 35 days show histopathological changes, a marked increase in weight and a lowering in the ascorbate content. A focal increase in the neutral lipid and cholesterol content was noted mostly in the inner cortical zones; and a characteristic acid phosphatase-positive pattern in areas of infiltrating cells, and an alkaline phosphatase-positive reaction in heterophils in the infiltrated areas. Electron microscopy confirmed that the zona glomerulosa cells were relatively normal in hypercholesterolaemic rabbits, while necrosis and fibrosis were very obvious in the inner two zones. The cellular infiltrate was shown to consist of large, granular mononuclear cells, heterophils, eosinophils, stromal phagocytes, lymphocytes and plasma cells. The possibility that the reaction was of an immunological nature is considered. The morphology of the adrenals of rabbits which were on a cholesterol-rich diet for 35 days and on a normal diet for 6 weeks afterwards, was indistinguishable from that of those rabbits killed after 35 days on a cholesterol-rich diet.

Posterior dislocation of the hip while playing basketball.

Injuries in basketball are usually to the ankles and knees. Dislocation of the hip is usually associated with severe trauma--for example, road traffic accidents. A case is reported here in which a 22 year old club basketball player slipped on landing from a jump shot, forcing him into a side splits position from which he sustained a posterior dislocation of the hip resulting in a sciatic nerve palsy.

The effect of paraquat on the in vitro activity of cytosol, mitochondrial and microsomal enzyme systems.

Subcellular fractions (mitochondria, microsomes and cytosol) were prepared from the lungs of rabbits and rats to investigate the effects of paraquat (Aldrich Laboratories) on the activity of some cytosol and mitochondrial dehydrogenases and on the microsomal respiration and reduced pyridine nucleotide oxidation rate. The normal basal oxygen consumption of rabbit lung microsomes was 1,9 +/- 0,3 nmol O2/mg microsomal protein/min, and the oxidation rates of reduced nicotinamide-adenine dinucleotide phosphate (NADPH) and reduced nicotinamide-adenine dinucleotide (NADH) were 4,29 +/- 0,53 and 4,0 +/- 0,55 nmol/mg microsomal protein/min respectively. One molecule of oxygen can therefore oxidize two molecules of NADPH or NADH, and the generated hydrogen peroxide is probably immediately broken down by the catalase activity of the normal lung microsomal preparation. When Aldrich paraquat (1,0 mM) was added to microsomes metabolizing NADPH (0,5-0,75 mM), both the rate of oxygen consumption and the generation of nicotinamide-adenine dinucleotide phosphate (NADP) were significantly (P less than 0,001) stimulated over the first 5 minutes, and thereafter returned to within basal limits. When microsomes were preincubated with 1,0 mM paraquat before NADPH was added, the oxygen consumption was substantially lower (10,01 +/- 1,01 nmol oxygen/mg microsomal protein/min), while the NADPH oxidation rate was almost similar to the basal rate in the absence of paraquat. This resulted in a striking dissociation in the H/O ratio under these circumstances. The addition of potassium cyanide (KCN) (5.0 mM) prior to paraquat pre-incubation and followed by the addition of NADPH restored the stimulatory effect of paraquat on microsomal respiration and on NADPH oxidation rate.(ABSTRACT TRUNCATED AT 250 WORDS)

Experimental paraquat poisoning--histological, electron microscopic and autoradiographic changes in the lung.

Paraquat is a potent and widely used herbicide which acts as a specific pulmonary toxin and causes lung fibrosis in man and animals. Some controversy still exists concerning the details of the morphogenesis of the pulmonary lesions. The lungs of rats exposed to intravenous injections of paraquat and sacrificed 6-24 days later were examined by light and electron microscopy. Autoradiography was used to detect possible paraquat accumulation in the lung 5 hours after a single intravenous injection. The findings on microscopy suggested an acute phase of damage to alveolar lining epithelium followed by epithelial regeneration. The most pronounced light and electron microscopic findings were: signs of disruption of the alveolar wall; type II alveolar epithelial hyperplasia; mobilization of mononuclear cells, and migration and accumulation of fibroblast-like cells in the intra-alveolar and interstitial spaces. After three equally spaced intravenous injections of paraquat signs of interstitial connective tissue proliferation could be seen. Autoradiography showed low-grade radioactivity over the alveolar wall, indicating possible active uptake of paraquat by alveolar epithelium; this coincides with in vitro evidence of an active transport mechanism for paraquat by alveolar epithelial cells.

Cyclosporin as a treatment for interstitial lung disease of unknown aetiology.

Ten patients with progressive, symptomatic interstitial lung disease of unknown aetiology who were treated with cyclosporin A were reviewed. Five had clinical and histopathological features of cryptogenic fibrosing alveolitis and five a progressive restrictive lung disease characterised by interstitial infiltration with lymphocytes and minimal fibrosis, which could not be classified precisely. Three patients with lymphocytic infiltration showed a response to initial treatment with cyclosporin A alone at high dosage, but toxicity precluded further treatment. All 10 patients then received low doses of cyclosporin A and prednisone. Three of the patients with cryptogenic fibrosing alveolitis and all five patients with lymphocytic infiltration responded with a reduction in dyspnoea or an increase in vital capacity, or both; cyclosporin A appeared to be effective, or at least to have a corticosteroid potentiating effect. A high incidence of side effects occurred, though these do not necessarily prohibit the long term use of cyclosporin A when it is indicated clinically. Cyclosporin A may be effective in the treatment of interstitial lung disease of unknown aetiology. Further studies are required to determine the long term outcome of treatment.

Lupus nephritis. Part I. Histopathological classification, activity and chronicity scores.

Renal biopsy has made a major contribution to the understanding and management of patients with lupus nephritis. In a 5-year retrospective study the renal morphology of 55 biopsies from 51 patients with lupus nephritis was classified according to World Health Organisation criteria. In addition, semi-quantitative activity and chronicity scores were documented. The findings were similar to series from other parts of the world. Of the biopsies reviewed, 6 were class II, 13 class III, 32 class IV and 4 class V. In situations of overlap, segmental proliferative features determined the class to which a biopsy specimen was assigned. Twenty-five of the patients, all WHO class IV, showed activity scores in the severe range. Most of the activity score features were common and easily recognised but necrotising angiitis was only seen in 1 patient. Haematoxylin bodies were difficult to document and the nature and value of the haematoxylin body is questioned.