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Most respondents felt that child care workers' legal concerns should be addressed by requiring (1) thorough, legally approved written procedures and guidelines for all day-to-day procedures and responsibilities; (2) legally approved comprehensive record-keeping systems; and (3) access to legal support and counsel. When child care workers follow legal guidelines and a facility's procedure manuals carefully, documenting their activities in an approved record-keeping system, they can expect to be in a positive situation in any legal contest and to have behavior measured from a legal standpoint by a standard described as what a "reasonable person" would be expected to do in a similar situation and circumstance.

The activity of serum angiotensin converting enzyme (ACE), assayed fluorometrically with the substrate hippuryl-L-histidyl-L-leucine was significantly higher in 116 patients with sarcoidosis than in 415 patients with other diseases--pulmonary and nonpulmonary--and in 58 normal subjects. Serum ACE was elevated in 59% of 46 recently diagnosed, untreated patients with sarcoidosis and was significantly reduced following steroid therapy in three patients with sarcoidosis, but in only one of four patients treated with placebos. The results suggest that elevated serum ACE activity is a valuable, although not absolutely specific, diagnostic indicator of sarcoidosis. It should be stressed that a normal value does not rule out the disease, and that steroid therapy tends to reduce the serum ACE activity.

Serum angiotensin-converting enzyme (SACE) was significantly elevated in 16 of 48 patients with various types of chronic renal disease not on hemodialysis [45.6 +/- (SD) 16.7 nmol/min/ml, p less than 0.001] and in 17 of 52 patients with chronic renal disease on maintenance hemodialysis (43.2 +/- 13.8, p less than 0.001) in comparison with 58 healthy adult controls (32.2 +/- 9.8). There was no significant relationship between SACE and renal function as indicated by serum creatinine and creatinine clearance, or the presence or absence of maintenance hemodialysis, except for a transiently increased elevation of SACE immediately post-dialysis due to hemoconcentration. There was a statistically insignificant positive correlation between SACE and 24-hour urinary protein excretion (r = 0.254, n = 21). Mean blood pressure measured in patients on maintenance hemodialysis was inversely related to SACE (p less than 0.05) following dialysis. Chronic renal disease irrespective of severity and therapy tends to be associated with elevated SACE and must be considered in the evaluation of sarcoidosis.

Serum collagenase was significantly elevated in stage II, but not stage I sarcoidosis, in contrast to elevation of serum angiotensin-converting enzyme in stages I and II. Neither serum collagenase nor angiotensin-converting enzyme was elevated in active tuberculosis. Elevated serum collagenase and angiotensin-converting enzyme levels were decreased with steroid therapy. Serum collagenase and angiotensin-converting enzyme were significantly correlated but did not vary identically. Collagenase was not elevated in lymph nodes in sarcoidosis, in contrast to marked elevation of angiotensin-converting enzyme. The results suggest that serum angiotensin-converting enzyme is a more sensitive index of sarcoidosis activity than serum collagenase, which may have an ancillary role in assessment of the disease.

To enable its immunohistologic localization, angiotensin converting enzyme (EC 3.4.15.1) from human lung was solubilized by trypsinization and purified approximately 2,660-fold to apparent homogeneity from a washed lung particulate fraction. The specific activity of pure enzyme was estimated to be 117 mumol/min per mg protein with the substrate hippuryl-l-histidyl-l-leucine. Consistent with previously described lung enzyme studies, catalytic activity was strongly inhibited by EDTA, O-phenanthroline, SQ 20,881, and SQ 14,225 and increased by CoCl(2). SQ 20,881 was a somewhat more potent inhibitor than SQ 14,225, unlike rabbit lung enzyme. The Michaelis constant (K(m)) with hippuryl-l-histidyl-l-leucine was 1.6 mM. The molecular weight was estimated at 150,000 from sucrose density gradient centrifugation. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed a single polypeptide chain estimated at 130,000 daltons. Rabbit antibody to human lung enzyme was prepared by parenteral administration of pure angiotensin-converting enzyme in Freund's adjuvant. Rabbit antibody to human lung angiotensin-converting enzyme appeared to crossreact weakly with the rabbit enzyme and strongly inhibited the catalytic activity of the enzymes from human serum, lung, and lymph node. The specificity of the rabbit antibody and purity of the final human lung enzyme preparation was suggested by the single precipitin lines obtained by radial double immunodiffusion, and by the coincidence of enzyme catalytic activity and immunoreactivity on polyacrylamide gel electrophoresis, with both relatively pure and highly impure enzymes. Generally applicable sensitive analysis of acrylamide gels for immunoreactivity (and subsequently for any other activity) by use of intact gel slices in radial double immunodiffusion was devised. Human lung enzyme was very tightly bound to and catalytically active on anti-human enzyme antibody covalently bound to Sepharose 4B, and could not be readily dissociated without inactivation. Antibody to human lung angiotensin converting enzyme has permitted tissue localization of the enzyme, which appears to be clinically useful in diseases associated with abnormal abundance of angiotensin-converting enzyme in tissues, such as sarcoidosis.

We investigated the possibility that epithelioid cells in sarcoidosis contain abundant angiotensin-converting enzyme, because these cells are derived from endothelial cells, which normally contain the enzyme in abundance. Granulomas from 9 patients with sarcoidosis were examined for the presence of the endothelial cell marker, factor VIII, by immunofluorescence microscopy using monospecific anti-human factor VIII antibody. Strong factor VIII-specific fluorescence was detected in the endothelial cells but no fluorescence was observed in the epithelioid cells. These results do not support the origin of epithelioid cells in sarcoidosis from endothelial cells as an explanation for their abundance of angiotensin-converting enzyme.

In adult chronic non-neuronopathic (Type 1) Gaucher's disease significant (p less than 0.001) elevations of angiotensin converting enzyme in serum (93.3 +/- 14.8 nmol/min/ml; number elevated, 8/11; normal control 32.2 +/- 1.30, n = 58) and spleen (5.62 +/- 0.35 nmol/min/mg protein, n = 2; control, 0.431 +/- 0.101, n = 4) and serum lysozyme (15.6 +/- 3.37 mug/ml; number elevated, 4/5) were observed. The KM for hippuryl-L-histidyl-L-leucine of Gaucher (1.31 mM) and normal (1.23 mM) serum angiotensin converting enzyme were similar. The increased angiotensin converting enzyme (ACE) in Gaucher's disease may be related to the genetic defect resulting in increased ACE synthesis in Gaucher cells, or perhaps generally, while high lysozyme may reflect an increased body mass of reticuloendothelial cells. These enzyme elevations may be of use in suggesting the possible presence of Gaucher's disease and perhaps in assessing the magnitude of pathologic involvement.

A simple, rapid, highly sensitive and reproducible fluorimetric assay for angiotensin-converting enzyme in untreated serum using the natural substrate based on a similar assay with hippuryl-L-histidine-L-leucine is described. Angiotensin I (0.2 mM in 0.1 M potassium phosphate-30 mM NaCl, pH 7.5; 37 C) is converted to angiotensin II and L-histidyl-L-leucine, which is quantified fluorimetrically (excitation = 360 nm; fluorescence, 500 nm) by formation of a fluorescent adduct with O-phthaldialdehyde. L-Histidyl-L-leucine peptidase was also monitored in order to correct for significant activity, which was observed only once, in less than 1% sera. The mean value of serum angiotensin-converting enzyme in sera from 45 normal subjects was 4.69+/-0.194 (SEM)+/-1.30 (SD) nmol/min/ml serum, compared with 31.7+/-1.53 (SEM)+/-10.3(SD) with the substrate analog hippuryl-L-histidine-L-leucine. There was a high degree of correlation between the velocity of cleavage of angiotensin I and hippuryl-L-histidyl-L-leucine (r - 0.903 to 0.993). The assay of serum angiotensin-converting enzyme is of use in the diagnosis and possible management of sarcoidosis and Gaucher's disease, and may have other applications.

A statistically highly significant elevation of serum ACE was found in a group of 58 patients with sarcoidosis (serum ACE was elevated in 34% of patients), as compared with normal controls and patients with tuberculosis and various other common diseases. The results suggest that serum ACE is a useful aid for the diagnosis of sarcoidosis when elevated, but that a normal value does not rule out the condition and may occur in more than one-half of monitored patients. There is a trend to diminution of serum ACE with increasing duration of disease with or without steroid therapy, perhaps correlating with the total body mass of active granulomas, as indirectly suggested in preliminary data by correlation of serum ACE with serum globulin in 16 sarcoidosis patients. It is not yet clear whether there is any significant steroid effect on serum ACE, but a significant number of patients on steroid therapy for more than 2-4 yr have elevated serum ACE values, which in some instances are extremely high. There was a 12-fold elevation in ACE to specific activities generally exceeding those of normal lung in granulomatous lymph nodes of 14 patients with sarcoidosis, suggesting that sarcoid granulomas may be actively synthesizing ACE and resulting in elevation of serum ACE. Extensively fibrotic sarcoid lymph nodes had normal or slightly elevated ACE, suggesting that obliteration of granulomas in sarcoid lymph nodes diminishes their ACE content and that this obliteration may be related to the tendency to diminution of serum ACE with time. ACE was not elevated in one tuberculous lymph node or in experimental granulomas, suggesting that elevation of ACE may have some specificity for the granuloma of sarcoidosis rather than being a characteristic of all granulomas. The catalytic and physical properties of ACE in serum and lymph nodes in sarcoidosis were generally similar to normal ACE with respect to pH activity, modulators, polyacrylamide-gel electrophoresis, and Sephadex G-200 gel filtration. However, sarcoid lymph node ACE appeared to be more heat labile than normal lung or lymph node ACE, suggesting the possibility that an abnormal ACE may be present in sarcoidosis. If an abnormal enzyme is indeed present, it might be coded for by a host gene that is not normally expressed or a nonhost gene or it might be a normal ACE that has been altered. No ACE activity was found in circulating white blood cells in sarcoidosis or in control subjects, suggesting that circulating white blood cells may not contain the epithelioid cell precursor or that ACE synthesis (or less likely, uptake) may be turned on at a later stage in the transformation. Lysozyme activity was also elevated in sarcoid lymph nodes. Serum ACE and serum lysozyme were significantly positively correlated in 16 sarcoidosis patients, suggesting a relationship between the two...

Hairy cells of hairy cell leukaemia are unique circulating mononuclear cells of questionable origin which possess features of both B-lymphocytes and monocytes. Angiotensin converting enzyme, which is consistently induced in monocytes in culture, was not induced in hairy cells in culture. This result is consistent with a lymphocytic rather than a normal monocytic cell lineage for the hairy cell. The origin of other monocytic phenotypic characteristics remains to be explained.

Rabbit serum, lung and corticosteroid-induced macrophage angiotensin-converting enzymes were compared with respect to migration on polyacrylamide-gel electrophoresis, sucrose gradient centrifugation and Km. Cellular particulate enzymes solubilized by nonidet P40 had approximately half the electrophoretic mobility of soluble enzymes and a similar Km (1.2 mM). Trypsin treatment of nonidet P40 solubilized particulate enzyme converted its electrophoretic mobility to that of soluble enzyme, and rendered it non-aggregating in sucrose gradients lacking detergent, similar to soluble enzyme. Approximate molecular weights by sucrose gradient centrifugation were similar for all enzymes (135,000-158,000). The data suggest that lung and macrophage enzymes are similar and that cellular particulate enzyme may be convertible to soluble enzyme.

The elevated activity of serum angiotensin converting enzyme (ACE) in sarcoidosis was usually reduced during steroid therapy but remained above the normal value (52 nmol-min-1-ml-1) in approximately a third of patients during the first year of therapy. ACE was elevated in 5 of 10 patients treated with steroids for at least four years. It was significantly higher in untreated patients (placebo group), who were subsequently placed on steroid therapy because of clinical deterioration, than in those remaining on placebo therapy. In general ACE tended to diminish with increasing duration of disease. The results suggest that serial study of ACE may be useful in the clinical management of sarcoidosis, including assessment of the degree of activity, the need for steroid therapy, therapeutic result and the prognosis.

Dose calibrators are not usually used to measure the activity of pure beta-emitting radionuclides. In this work, the activity of 32P-sodium phosphate was accurately measured with a Capintec CRC-2 dose calibrator. Using a calibration knob setting of 012 on the 1-mCi range, the 32P dose (in mCi) could be calculated directly simply by multiplying the instrument readout by 10. The dose calibrator response was found to be linear at this knob setting and moderate alterations in geometry produced no significant changes.

The relationship between serum angiotensin-converting enzyme activity and inflammatory bowel disease was investigated in 37 patients who had Crohn's disease, 31 patients who had ulcerative colitis, and 104 control subjects. The enzyme activity tended to be depressed in Crohn's ileitis (P less than 0.05) and colitis, but not in Crohn's ileocolitis and ulcerative colitis. No increase in enzyme activity was observed in Crohn's disease ileum or colon or in ulcerative colitis colon. The granulomatous inflammation in Crohn's disease differs from that in sarcoidosis, in which striking elevation of angiotensin-converting enzyme is present in granulomatous tissue and frequently in serum.

Monoclonal antibodies have begun to assume a significant role in clinical research. The ability to label these agents has initiated research in the areas of radioimmunodetection and radioimmunotherapy. In the case of antibodies directed against tumor antigens, imaging has been employed to help assess location and extent of disease, and to provide information and extent of disease, and to provide information concerning biodistribution to be used in subsequent dosimetric calculations. Because of the low counting statistics characteristic of such images, the use of single photon emission computed tomography (SPECT) is suggested as a potential method of improving the diagnostic yield from image data. Careful attention to acquisition parameters and image processing options is needed if these goals are to be achieved.

Angiotensin-converting enzyme (ACE) was detected by indirect immunofluorescence examination. ACE was present in the cytoplasm of epithelioid cells in granulomas from 38 of 39 patients who had clinical sarcoidosis but not in any of 37 nonsarcoid granulomas, normal lymph nodes, or splenic controls. Intensity of staining with ACE antiserum was inversely proportional to the degree of fibrosis and was significantly less intense in fibrotic granulomas. The detection of ACE appears to be a distinct aid in establishing a definitive tissue diagnosis of sarcoidosis.

Radioimmunotherapy retreatment of patients receiving radiolabeled murine monoclonal antibodies is difficult because of human antimurine antibody (HAMA) formation. Retreatment therapy was initiated in three patients at the time of disease progression using a radioiodinated monoclonal antibody (T101). The clinical protocol consisted of a two day plasma exchange (4-6 L) to reduce HAMA titers. Immunoimaging was performed with 5 mCi 131I-T101 (10 mg). Gamma scintillation images were obtained 18 hr postinfusion, and radiation dosimetry estimates were performed. At 24 hr postinfusion, each patient received a 100-mCi 131I-T101 (10 mg) therapy dose. Results obtained after plasmapheresis showed a significant reduction, ranging from 28%-61%, in HAMA titers. Blood clearances were markedly different between initial therapy and retreatment therapy for patient with high HAMA titers, reflecting immune complex formation. Two patients responded to retreatment therapy with responses lasting 1 to 2 mo. Minimal acute and no chronic toxicities were observed during the retreatment protocol.

Low angiotensin-converting enzyme (ACE) activity was found in rat and mouse peritoneal macrophages, in rat and rabbit pulmonary alveolar macrophages, in cultured mouse peritoneal macrophages stimulated i.p. for four days by thioglycollate, and in rabbit pulmonary alveolar macrophages stimulated in culture by 0.1 to 100 microgram/ml Salmonella typhosa endotoxin for four days. Rat subcutaneous Freund's adjuvant granulomas induced with Mycobacterium butyricum or M. tuberculosis H37 Ra contianed low ACE activity which was generally lower than that present in control tissues. Increased ACE activity in sarcoidosis and Gaucher's disease lesions does not reflect a universally high synthesis of ACE in mammalian macrophages or granulomas, but may be due to specific mechanisms.

In this study, an IgM monoclonal antibody (MAb600D11) directed against human small cell lung cancer (NCI-H69) was radiolabeled with iodine-131, and the biodistribution and image quality of the radiolabeled antibody was evaluated. Radiolabeling was achieved in a solid-phase system consisting of 1,3,4,6-tetrachloro-3a,6a-diphenylglycoluril. Labeling efficiencies and protein purification were accomplished using gel exclusion chromatography while radioimmunoreactivity was determined using a solid-phase radioimmunoassay procedure. The biodistribution of I-131-labeled MAbs was determined in Sprague-Dawley rats up to 7 days after injection. Highest organ concentrations were observed in liver (3.91 +/- 0.47 (SD) and 0.17 +/- 0.04 (SD) mean percent injected dose at 1-7 days after injections) and in thyroid (5.33 +/- 0.71 (SD) and 5.32 +/- 2.01 (SD) mean percent injected dose at 1-7 days after injection). Nude mice, bearing either a small cell lung tumor (NCI-H69) or a nonspecific tumor (adenocarcinoma), were injected with 400-800 microCi of I-131 labeled monoclonal antibody. Optimum tumor visualization was observed 2-4 days after injection with tumor concentrations as high as 10.4% of the initial injected dose. The results demonstrated that radioimmunoimaging of human small cell lung carcinoma was feasible with the tumor-specific IgM I-131-labeled MAb.

A simple, rapid, highly sensitive and reproducible assay for angiotensin-converting enzyme in untreated serum is described. It is based on the conversion of the substrate analog, hippuryl-L-histidyl-L-leucine (5 mM in 0.1 M K phosphate, pH 8.3-0.3 M NaCl) to hippurate and L-histidyl-L-leucine, which is quantified spectrofluorimetrically (lamdba excitation = 360 nm; lamdba fluorescence = 500 nm) by formation of a fluorescent adduct with ophthaldialdehyde. The chloride requirement and inhibition and activation patterns correspond to those for angiotensin-converting enzyme. The Km for hippuryl-L-histidyl-L-leucine was 1.33 mM. The mean value of serum angiotensin-converting enzyme for 58 normal human subjects (mean age, 32 years; range 19-57) was 32.2 +/- 1.30 (SE), with a standard deviation of 9.87 nmol/min/ml serum. The assay is useful for the diagnosis and possible management of sarcoidosis and may have other applications in the future.

Serum angiotensin-converting enzyme was elevated in patients with hyperthyroidism (72 +/- 31 nmol/minute/ml, n = 12, p less than 0.001) but not in patients with hypothyroidism (38 +/- 3, n = 3) or thyroiditis (26, n = 1), and was positively correlated in 23 patients with serum thyroxine concentration (r = 0.60, p less than 0.01) and triiodothyronine resin uptake (r = 0.56, p less than 0.01). Triiodothyronine failed to enhance the synthesis of angiotensin-converting enzyme in rabbit alveolar macrophages or in human monocytes in culture, suggesting that the increased serum enzyme is a consequence of an effect other than increased angiotensin-converting enzyme synthesis. Hyperthyroidism should be considered in the evaluation of serum angiotensin-converting enzyme for the diagnosis and management of sarcoidosis.

To determine whether elevated thyroid hormone is responsible for increased serum angiotensin converting enzyme in hyperthyroidism, 5 to 40 micrograms of 3,5,3'-triiodo-L-thyronine was administered orally and subcutaneously to female Swiss-Webster mice. Serum angiotensin converting enzyme was significantly increased in all animals given triiodothyronine compared to controls. Lung and kidney enzymes were moderately reduced in specific activity but unchanged in total activity due to increase in size of these organs. The results indicate that in hyperthyroidism, elevated thyroid hormone per se rather than the disease of the thyroid is responsible for elevated serum angiotensin converting enzyme.

Serum angiotensin-converting enzyme in a patient with type 2 acute neuronopathic Gaucher's disease (242 nmol/min/ml) was 10.8 times higher than values for eight patients with other hereditary neurologic abnormalities (22.5 +/- 2.0) and 9.4 times higher than those for 12 patients with other diseases (25.7 +/- 2.6) (P less than 0.001). Serum lysozyme was not elevated in the patient with type 2 Gaucher's disease. These results indicate that elevated serum angiotensin-converting enzyme in an infant with neurologic involvement and hepatosplenomegaly is suggestive of the possibility of type 2 Gaucher's disease. Typical Gaucher's cells and fibrosis were observed by light and electron microscopy of the liver. An aspect hitherto unreported in Gaucher's disease or in the liver was that approximately 20% of the collagen fibrils were of the long-spacing type, with periodicity of 1,000 to 1,100 A and diameters of 900 to 1,500 A.

A new method of obtaining scintigraphic images of the cardiac blood pool for the study of ventricular wall motion is described. The method, which is based upon an inexpensive modification of a commercial multiformat imaging device, yields a set of serial gated images covering the entire cardiac cycle. These images may be converted into a film loop to yield a continuous motion picture of the mechanical cycle of the heart.