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Biomedical subjects

S F Ryan

Publications and source records attributed to S F Ryan.

At least 19 recordsLinked to original sources
PubMed1991

Purification of surfactant protein A from dog lung by reconstitution with surfactant lipids.

We have developed a simple method for purification of surfactant major apoprotein (SP-A, MW 34-38 kD) from dog lungs with high yield and purity. Lipids and proteins of partially purified surfactant were dissociated by sodium deoxycholate (DOC, 100 mM, 37 degrees C, 30 min), diluted 1:10 with borate buffer containing 3 mM CaCl2, and dialysate in the same buffer to reconstitute the lipids and proteins (4 degrees C, 48 h). The reconstituent and the partially purified surfactant were purified by ultracentrifugation on a discontinuous sucrose density gradient. Protein was isolated from the reconstituent and from the purified surfactant by delipidation, and the yields and purities were assessed by one-dimensional SDS-PAGE and 2-dimensional electrophoresis (isoelectric focusing, SDS-PAGE). We found that the surface pressure-time adsorption isotherm, minimum surface tension, and the yield and composition of lipids of the reconstituent were identical with those from the purified surfactant. Only about 0.25% of the DOC used for dissociation remained with the reconstituent and it did not affect surface properties of the reconstituent. The yield of SP-A in the reconstituent was almost the same as that in the purified surfactant, but the former contained no plasma protein whereas the latter contained significant amounts. The amino acid composition and the partial N-terminal amino acid sequence of SP-A were the same as those from the purified surfactant. Reconstituent prepared from surfactant lipids and SP-A adsorbed more rapidly and reached a higher final surface pressure than did the surfactant lipids alone. These results demonstrate that large quantities of SP-A can be purified by reconstitution with surfactant lipids, and that the purified protein is biophysically active.

Amino Acid Sequence↗
PubMed1991

Effects of activated polymorphonuclear leukocytes upon pulmonary surfactant in vitro.

Current evidence suggests that products of activated inflammatory cells cause or contribute to the acute lung injury of the adult respiratory distress syndrome (ARDS). To assess the possibility that these products may impair surfactant function during ARDS, we exposed surfactant in vitro to polymorphonuclear leukocytes (PMN) activated by phorbol myristate acetate and to the oxidant-producing pair ferric chloride/ascorbate (FeCl3/ASC). After incubation of surfactant with 8 to 32 x 10(6) activated PMN for 1 to 4 h or with FeCl3/ASC for 16 h, its isopycnic density (d), minimum surface tension (gamma min), time course of adsorption, compressibility (SC), and stability index (SI) were determined. We found progressive decreases of d, adsorption, and SI and progressive increases of gamma min and SC after exposure to activated PMN in increasing numbers or for longer time periods. Superoxide dismutase completely inhibited all of these effects except the decreased adsorption, which it did not significantly inhibit. Similar changes in all of these parameters occurred after exposure of surfactant to FeCl3/ASC. Polyacrylamide gel electrophoresis of surfactant after exposure to activated PMN showed a decrease of the major apoprotein that progressed with exposure time and was associated with the appearance of several bands with both lower and higher molecular weights than that of the apoprotein. The data show that activated PMN are capable of impairing surfactant function in vitro and of degrading the major apoprotein. They suggest that the effects upon d, gamma min, SC, and SI are mediated largely if not exclusively by oxidant radicals. While oxidants may contribute to delayed adsorption, proteolysis appears to play the principal role in this effect.

Adsorption↗
PubMed1988

The distribution of atherosclerotic lesions in the coronary arterial tree: relation to cardiac risk factors.

Although it is often stated that proximal atherosclerotic coronary artery disease occurs more frequently than distal disease, several autopsy studies have disputed this. To examine the prevalence of proximal vs mid and distal disease and its relationship with cardiac risk factors, we studied more than 14,000 sections from 102 hearts with coronary artery disease at autopsy. After postmortem angiography, the coronary arteries were removed, divided into proximal, mid, and distal thirds, sectioned at 2.5 mm intervals, and graded for percentage reduction in cross-sectional area by atherosclerosis. Of 252 vessels in 84 patients with greater than or equal to 75% stenosis, 166 (66%) has proximal disease vs 107 (42%) with mid disease and 40 (16%) with distal disease (p less than 0.001). No patient had a mid or distal stenosis greater than 75% without proximal disease. When atherosclerosis of any severity was assessed, proximal atherosclerotic lesions were long and diffuse, whereas distal lesions were more often short and discrete. Proximal circumflex lesions were shorter in length than those in the right or left anterior descending coronary arteries. The prevalence of proximal, mid, and distal stenoses in 25 diabetic patients was similar to that in nondiabetic persons (53%, 47%, and 17%, p greater than 0.3). Similarly, hypertension, smoking, and obesity were not associated with an increase in prevalence of distal disease. Patients with distal stenoses were younger than patients without (mean age, 64 +/- 13 vs 73 +/- 10 years, p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗
PubMed1988

Human pulmonary dirofilariasis: does diagnosis require thoracotomy?

The incidence of pulmonary coin lesions as a result of Dirofilaria immitis (dog heartworm) seems to be increasing. A case of human pulmonary dirofilariasis is described, and its pathogenesis and the limitations of preoperative diagnostic tests in this condition are discussed.

Adult↗
PubMed1987

Glandular cardiac myxomas. Histologic, immunohistochemical, and ultrastructural evidence of epithelial differentiation.

Histologic, histochemical, immunocytochemical, and ultrastructural features of two cardiac myxomas containing glandular elements are reported. Glandular elements in both cases stained positively with both mucicarmine and periodic acid-Schiff reagent with diastase pretreatment (DPAS). Immunoperoxidase studies demonstrated positivity of the glandular cells for carcinoembryonic antigen (CEA), epithelial membrane antigen (EMA), and keratin. Factor VIII-related antigen (FVIIIAg) was identified only in cells lining vascular spaces. Electron microscopic study of one tumor demonstrated well-formed glands having basement membranes, junctional complexes, and apical secretory granules. These findings indicate the capacity for true epithelial differentiation of cardiac myxomas and have implications both as regards the histologic diagnosis of these tumors and their histogenesis.

Adult↗
PubMed1987

Functional abnormalities of lung surfactant in experimental acute alveolar injury in the dog.

Acute alveolar injury (AAI) was induced in dogs by injection of N-nitroso-N-methylurethane. Two to 20 days after injection, alveolar lavage phospholipids were quantified. Lavage surfactant was partially purified by centrifugation (27,000 g for 2 h), and further purified by centrifugation in NaBr density gradient (100,000 g for 4 h). Phospholipids, neutral lipids, surfactant-associated proteins, and surface properties of partially purified and purified surfactants were analyzed. Lavage disaturated phosphatidylcholine (DSPC) decreased to 37% of control at peak injury (Days 6 to 8) and increased to near normal during recovery (Days 10 to 20). Lavage phosphatidylglycerol (PG) decreased to 22% of control at peak injury and remained in that range through recovery. In both partially purified and purified surfactants, percentages of phosphatidylcholine (PC), DSPC, phosphatidylethanolamine, and cholesterol in all phases of injury and recovery were not different from those in control animals. However, percentage of PG decreased markedly during injury and remained low through recovery, whereas those of phosphatidylinositol and lysoPC increased with injury and remained elevated through recovery. The PC-to-sphingomyelin ratio (L/S ratio) and percentage of triglyceride decreased during injury and returned to control values during recovery. Surfactant apoprotein of molecular weight 38,000 from partially purified and purified surfactant decreased markedly at peak injury and recovered to normal during recovery. During early and peak injury, both preparations failed to reduce surface tension below 19 dyne/cm and their isopycnic densities were altered. These studies indicate that, in addition to decreased quantity, qualitative changes in lipids and apoproteins and reduced surface activity of the surfactant occur during nitrosourethane-induced AAI.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Disease↗
PubMed1985

Normal surface properties of phosphatidylglycerol-deficient surfactant from dog after acute lung injury.

Lung surfactant was isolated from bronchoalveolar lavage of dogs during the late phase of recovery (15 days) from acute alveolar injury induced by subcutaneous injection of N-nitroso-N-methylurethane. This surfactant was compared with surfactant from control dogs in terms of in vitro surface properties, phospholipid composition and protein content, and those of its subfractions. Phospholipid composition and protein content were similar in the two groups, except that phosphatidylglycerol (PG) was markedly reduced and phosphatidylinositol (PI) was increased in the experimental group. In both, isopycnic densities of their subfractions in continuous sucrose density gradient were identical. The time course of surfactant adsorption was similar in both groups. Minimum surface tension (gamma min) was 4.1 +/- 1.5 dynes/cm in the experimental dogs and 3.8 +/- 1.3 dynes/cm in the controls. Surface compressibility (SC), stability index (SI), and dynamic respreadability (DR) of the surfactants from the two groups were nearly identical. When compared to an artificial surfactant composed of dipalmitoyl phosphatidylcholine (DPPC) and PG in 9:1 molar ratio a mixture of DPPC-PI 9:1 prepared identically showed similar gamma min, SC, SI, and DR, and a much higher surface adsorption rate. These results suggest that PG is not essential for normal in vitro surfactant function and that its role may be assumed by PI.

Animals↗
PubMed1984

Diphosphatidylglycerol in experimental acute alveolar injury in the dog.

Acute alveolar injury closely resembling that seen in humans was induced in dogs by subcutaneous injection of N-nitroso-N-methylurethane. Necrosis of alveolar epithelial cells was observed during early injury. Proliferation of immature epithelial cells which began during early injury and became massive after peak injury was followed by their differentiation to mature type II cells during recovery. Quantities of diphosphatidylglycerol (DPG) and of phosphatidylglycerol (PG) in alveolar lavage and in post-lavage lung tissue were measured. An increase in tissue DPG coincided with a sharp decrease in tissue and lavage PG during early injury. DPG was not detectable in the lavage. During late recovery, tissue DPG increased threefold over controls. This increase was accompanied by persistence of a 50% decrease in tissue PG and 83% decrease in lavage PG. Biosynthesis of DPG and PG in isolated lung mitochondria demonstrated that DPG was formed from PG in the presence of CDP-diglyceride. These findings suggest that the low level of PG in the surfactant complex during acute alveolar injury is due to increased turnover of PG to DPG in the lung.

Animals↗
PubMed1979

Adipose cell morphology and control of lipolysis in a patient with partial lipodystrophy.

A previously unreported patient with partial (cephalothoracic) lipodystrophy is described. Glucose tolerance and plasma lipids were normal, but plasma insulin increased to 340 muU/ml during an oral glucose tolerance test. Plasma free fatty acids were appropriately suppressed by oral glucose, insulin, and nicotinic acid, and were increased by infusion of norepinephrine. The lipolytic responses was also normal in response to two stimuli for endogenous catecholamine release: upright posture and 2-deoxyglucose infusion. There was a gradual development of postural hypotension in response to upright posture despite appropriate reflex tachycardia. Anhidrosis was present over the lower half of the body during this test, in a distribution corresponding to the area of adipose tissue hypertrophy. Anhidrosis was also seen in this region in response to warm ambient temperature. Adipose cells from the atrophic area were smaller than those from the hypertropic area, but the atrophic cells were only 65% of the volume of the hypertrophic cells by two different methods. Thus, loss of cells occurred. Glucose-1(-14)C utilization and in vitro lipolysis were similar in the two cell preparations; the difference were explicable by cell size and did not suggest a metabolic abnormality. Counts of unmyelinated nerves were similar in the two areas. These findings indicate that in this patient the lipodystrophy was associated with normal fat cells and an autonomic dysfunction. However, the findings cannot completely explain the pathogenesis of her disorder. Loss of fat cells, rather than symmetrical shrinkage, occurred in the upper half of the body.

Adipose Tissue↗
PubMed1979

Alveolar epithelial injury causing respiratory distress in dogs: physiologic and electron-microscopic correlations.

Structural and mechanical changes were correlated in 29 dogs with acute alveolar injury induced by the subcutaneous administration of N-nitroso-N-methylurethane (NNNMU). The injury was characterized by necrosis and repair of alveolar epithelium while the vascular endothelium remained essentially intact. Compliance of the lung (CL) decreased and elastic recoil increased as epithelial necrosis occurred. During recovery, improved elastic recoil coincided with epithelial regeneration, although CL remained abnormal. The late phase was characterized histologically by widespread closure of clusters of alveoli alternating with dilated small air spaces. The process resulted in distortion of lung architecture and resembled interstitial fibrosis. Reduced lung volume and decreased distensibility of dilated small air spaces may have accounted for the persistently abnormal CL. Because of the specific site of anatomic involvement, the predictable evolution of deranged lung mechanics, and the similarity to human lung injury (adult respiratory distress syndrome), the lung injured by administration of N-nitroso-N-methylurethane is a suitable model to study pathophysiology and types of therapy in a controlled setting.

Animals↗
PubMed1979

Steatosis of granular pneumocytes in alcoholics with acute alveolar injury.

Accumulation of neutral lipid in the type II alveolar epithelial cells of the lung has been described in experiments involving animals with conditions such as hypoxia or on alcohol administration. In two cases involving human subjects, this change was observed at autopsy by histochemical stains and electron microscopy. In both instances, the patients had had severe alcoholic liver disease, as well as extreme hypoxia resulting from acute alveolar injury. The lungs of six alcoholic patients with liver disease but without acute alveolar injury showed no lipid vesicles on histochemical staining. These observations suggest that a metabolic insult or combination of insults, such as alcohol or hypoxia, might lead to accumulation of neutral lipid, especially in regenerating alveolar epithelial cells that may be more susceptible to such injury.

Alcoholism↗
PubMed1978

Volume-pressure and morphometric observations after acute alveolar injury in the dog from N-nitroso-N-methylurethane.

Volume-pressure diagrams during inflation with air and saline were made with the lungs of 6 control dogs and 24 dogs with acute alveolar injury induced by subcutaneous injection of 6 mg of N-nitroso-N-methylurethane per kg of body weight 3 to 14 days before study. The extent of alveolar closure was estimated by measuring the mean linear intercept of the remaining open air spaces after inflation of the lung with liquid formalin at a pressure of 40 cm H2O. This alveolar closure was defined as irreversible. The volume-pressure diagrams and compliance data derived from them during the 3 to 4, 5 to 7, and 9 to 14 day periods after injection were analyzed and compared with the morphometric data. The diagrams with air inflation showed a progressive downward shift beginning with the 3 to 4 day period. This shift was at least partly independent of volume loss. The diagrams with saline inflation were unchanged during the 3 to 4 day period, but showed a downward shift, largely due to volume loss thereafter. Irreversible alveolar closure, reflected by an increased mean linear intercept, was present in the 3 to 4 day period, and its extent increased with time, correlating inversely with total lung capacity and saline compliance. The data indicate that decreased lung compliance and volume during the first 7 days of injury induced by injection of N-nitroso-N-methylurethane were due largely to increased surface tension and that tissue forces became a significant factor later. They suggest that the altered tissue forces were due principally to irreversible alveolar closure.

Animals↗
PubMed1977

Adult respiratory distress syndrome.

The authors correlated and presented clinical, radiological and pathological findings in 6 patients with Adult Respiratory Distress Syndrome (ARDS): It is a distinct syndrome with characteristic clinical, radiological presentations and pathological findings. It is a unique response of lung to a variety of primary insults. If unrecognized, patients with this syndrome are often fatal. Its possible pathognesis is discussed.

Adult↗
PubMed1976

Experimental acute alveolar injury in the dog. Morphologic--mechanical correlations.

In 26 dogs, a single subcutaneous injection of N-nitroso-N-methylurethane produced acute lung injury characterized by tachypnea cyanosis, increased static lung recoil, and decreased lung compliance. During the first few days, light microscopic examination revealed widespread interstitial and perivascular edema and alveolar collapse. At the same time, electron microscopy showed the major alteration to be widespread necrosis of both types of alveolar epithelial cells without significant injury to the vascular endothelium. During recovery, new epithelial cells appeared which probably were derived from granular pneumocytes. These cells developed into mature granular pneumocytes through a phase in which they resembled fetal granular pneumocytes. The late stage was characterized by a picture resembling diffuse interstitial fibrosis but which was due to irreversible closure of clusters of small airspaces with no apparent increase in collagen. Elastic recoil of the lungs, as reflected by peak inspiratory airway pressure, increased during the acute phase and showed a return toward normal that was coincident with the appearance of mature granular pneumocytes in the regenerating epithelium. Lung compliance decreased during the acute phase and in most animals returned toward normal during the recovery phase. These observations strongly suggest that the alteration in lung mechanics is related to epithelial necrosis and that recovery is related to epithelial regeneration.

Acute Disease↗