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A E Broadus

Publications and source records attributed to A E Broadus.

At least 73 records · Page 4Linked to original sources
PubMed1987

Two forms of parathyroid hormone-like adenylate cyclase-stimulating protein derived from tumors associated with humoral hypercalcemia of malignancy.

Tumors associated with humoral hypercalcemia of malignancy (HHM) contain parathyroid hormone-like adenylate cyclase-stimulating proteins (hACSPs). We previously purified a 17,000 MW hACSP from an HHM-associated breast carcinoma. This report describes the characterization of hACSPs from three additional HHM-associated tumors: two typical HHM-associated tumors (squamous carcinomas) and a third unusual tumor type (pheochromocytoma). Each tumor was extracted in acid-urea/ethanol-sodium chloride, and adenylate cyclase-stimulating activity (ACSA) was examined following reverse-phase and size-exclusion HPLC and isoelectric focusing. Each tumor contained a high molecular weight form of ACSA which co-eluted with the 17,000 molecular weight breast carcinoma hACSP in each of the three separation procedures. Each also contained a lower molecular weight form(s) of ACSA (6,000-9,000 molecular weight). Both forms were inhibited by Nle8,18Tyr34bPTH(3-34) amide. The high molecular weight form was not changed to the lower molecular weight form by a reducing agent. Some HHM-associated tumors contain two forms of hACSP, one with a molecular weight of 17,000 and another with a molecular weight of 6,000-9,000, which appears to be an amino-terminal cleavage product of the larger protein.

Adenylyl Cyclases↗
PubMed1987

Hypercalcemia of malignancy.

Malignancy-associated hypercalcemia is a common and important clinical problem. Two major pathogenic mechanisms appear to be operative in this disorder: local osteolytic hypercalcemia and humoral hypercalcemia of malignancy. The clinical and biochemical features that distinguish these two processes are reviewed.

Aged↗
PubMed1987

Co-purification of transforming growth factor beta-like activity with PTH-like and bone-resorbing activities from a tumor associated with humoral hypercalcemia of malignancy.

Humoral hypercalcemia of malignancy (HHM) is caused by a circulating bone-resorbing factor or factors. Suggestions as to the nature of this factor include PTH-like proteins, transforming growth factors, and bone-resorbing factors distinct from either of the first two classes of polypeptides. We investigated the occurrence of these three activities in a highly purified extract of the H-500 Leydig cell tumor which causes HHM when implanted into Fisher rats. PTH-like adenylate cyclase-stimulating activity (ACSA) was extracted from tumor tissue by sequential treatment with urea/HCl and ethanol/NaCl. Tumor extract was further purified by hydrophobic-interaction, gel-filtration, and reverse-phase HPLC steps to a specific activity of 1038 ng eq bPTH(1-34)/mg protein. Only the fraction pool containing ACSA demonstrated significant bone-resorbing (1.78-fold over basal) and transforming growth factor activity (epidermal growth factor (EGF)-dependent colony formation in soft agar suspension by NRK-49F indicator cells). A subsequent reverse-phase HPLC step produced material which contained both ACSA and transforming growth factor beta (TGF beta)-like activity in a single fraction. Whether the responsible mediator in this animal model has TGF beta-like properties as well as PTH-like and bone-resorbing activity remains to be determined.

Adenylyl Cyclases↗
PubMed1986

A parathyroid hormone-like protein from cultured human keratinocytes.

Parathyroid hormone-like factors have been found in extracts of tumors associated with humoral hypercalcemia of malignancy, many of which are of squamous epithelial origin. Cultured, nonmalignant human keratinocytes were examined for the production of similar factors. Keratinocyte-conditioned medium from ten cultures stimulated the production of cyclic adenosine monophosphate in clonally derived rat osteosarcoma cells sensitive to parathyroid hormone. Bovine [Nle8,18, Tyr34]PTH-(3-34)NH2, a competitive inhibitor of parathyroid hormone, stopped the adenylate cyclase production stimulated by keratinocyte-conditioned medium, but antisera to parathyroid hormone had no effect on such adenylate cyclase activity. The active component of keratinocyte-conditioned medium has a molecular weight exceeding that of native parathyroid hormone. These characteristics are shared by the parathyroid hormone receptor agonists associated with humoral hypercalcemia of malignancy, which suggests that normal human keratinocytes may produce a factor related to that produced by malignant tumors associated with humoral hypercalcemia of malignancy.

Adenylyl Cyclases↗
PubMed1986

Frequency and partial characterization of adenylate cyclase-stimulating activity in tumors associated with humoral hypercalcemia of malignancy.

Parathyroid hormone-like adenylate cyclase-stimulating activity (ACSA) has previously been identified in small numbers of tumors or tumor-conditioned tissue culture medium derived from patients or animals with humoral hypercalcemia of malignancy (HHM). We examined the frequency with which this ACSA occurred in a large group of tumor extracts derived from patients with HHM (n = 20), and compared this to three control groups: normocalcemia-associated tumors (n = 20), hypercalcemic control tumors (n = 7), and normal, nonmalignant tissue samples (n = 10). Eighteen of 20 HHM-associated tumor extracts displayed ACSA whereas only 4 of 37 controls contained detectable ACSA. ACSA in one tumor was partially purified, using sequential extraction steps and reverse-phase, high-performance liquid chromatography. Highly purified ACSA (4800-fold) also contained potent in vitro bone-resorbing activity. The molecular weight as assessed by gel filtration was approximately 40,000 D. These findings provide strong support for the thesis that the humoral factor which is responsible for the syndrome of HHM is a parathyroid hormone-like adenylate cyclase-stimulating protein.

Animals↗
PubMed1986

Pregnancy as state of physiologic absorptive hypercalciuria.

An increase in circulating, 1,25-dihydroxyvitamin D level and net intestinal calcium absorption have been previously demonstrated in pregnant women and have been widely regarded as compensatory mechanisms whereby fetal mineral demands are satisfied. The alternate possibility, that these adjustments might anticipate such demands, has not previously been considered. To examine the effects of pregnancy on the intestinal absorption and renal excretion of calcium, oral calcium tolerance tests were performed and urinary calcium excretion was measured in 16 healthy women receiving a moderate calcium intake during and after pregnancy. Circulating 1,25-dihydroxyvitamin D levels and indexes of parathyroid function were also measured. As expected, 1,25-dihydroxyvitamin D levels were significantly (p less than 0.05) elevated throughout pregnancy (94 +/- 11, 118 +/- 9, and 117 +/- 11 pg/ml in the first, second, and third trimesters, respectively, versus 51 +/- 5 pg/ml after delivery). Twenty-four-hour calcium excretion also increased sharply (247 +/- 54, 316 +/- 42, 300 +/- 61 mg versus 91 +/- 18 mg), often to the point of hypercalciuria. Calcium tolerance test results included significant increases in the calciuric and calcemic responses during each trimester, whereas fasting calcium excretion and parathyroid function remained normal. These findings portray normal pregnancy as a state of physiologic absorptive hypercalciuria and call into question the widespread practice of supplementing calcium intake in otherwise well-nourished women during pregnancy.

Adult↗
PubMed1986

Two species of adenylate cyclase-stimulating activity in a murine squamous carcinoma model of humoral hypercalcemia of malignancy.

PTH receptor-stimulating proteins may be a common mediator of humoral hypercalcemia of malignancy (HHM). Such proteins exhibit adenylate cyclase-stimulating activity (ACSA) in PTH-sensitive assays, and ACSA has been used to follow their purification. Acid/urea tumor extracts from a murine squamous carcinoma model of HHM were previously shown to have very high ACSA, which was partially, but incompletely, inhibited by the PTH antagonist Nle8,18,Tyr34-bovine PTH-(3-34) amide. ACSA from murine tumor extracts has now been further purified using solvent fractionation and reverse phase HPLC. Approximately half of the ACSA is attributable to a family of three proteins (peaks IA, IB, and IC) with properties characteristic of the PTH receptor-stimulating protein extracted from rat Leydig cell and human HHM tumors. The ACSA in these three peaks of murine tumor extract elutes in the same region as human tumor ACSA on reverse phase HPLC, has a dose-response curve parallel to that of PTH, and is fully inhibited by the PTH-(3-34) antagonist in both the renal cortical and rat osteosarcoma (ROS) adenylate cyclase assays. The remaining half of the ACSA from murine tumor extracts elutes as a single peak (peak II) at a higher acetonitrile concentration on reverse phase HPLC. In the renal cortical assay, its dose-response curve differs from that of PTH, its ACSA is not affected by the PTH-(3-34) antagonist, and it potentiates PTH- or peak I-stimulated adenylate cyclase activity. In the PTH-sensitive intact cell ROS assay, peak II exhibits no ACSA. We conclude that the potent ACSA of murine tumor acid/urea extract results in large part from amplification of the PTH-specific ACSA (peak I) by peak II. Peak II is a distinct protein, not previously reported in tumor extracts, that may act as a postreceptor step in the adenylate cyclase system.

Adenylyl Cyclases↗
PubMed1985

Sensitivity of the parathyroid hormone-1,25-dihydroxyvitamin D axis to variations in calcium intake in patients with primary hyperparathyroidism.

Previous studies have demonstrated a spectrum of parathyroid responsivity to alterations in the extracellular calcium concentration in patients with primary hyperparathyroidism, but studies employing physiologic amounts of calcium have not, to our knowledge, been reported. We studied 18 unselected patients with primary hyperparathyroidism at the lower (400 mg) and upper (1000 mg) limits of a normal dietary intake of calcium. The diet containing high-normal amounts of calcium induced only a slight increase in 24-hour calcium excretion (from 281 to 337 mg per day) yet was associated with significant reductions in fasting serum levels of immunoreactive parathyroid hormone (from 60 to 50 nleq per milliliter; P less than 0.001), nephrogenous cyclic AMP (from 3.52 to 2.63 nmol per deciliter of glomerular filtrate; P less than 0.001), and plasma levels of 1,25-dihydroxyvitamin D (from 74 to 58 pg per milliliter; P less than 0.001). A wide spectrum of responses was observed, with some patients appearing to have essentially autonomous parathyroid function and others having marked suppressibility (up to 50 per cent) of the parathyroid hormone-vitamin D axis. We conclude that parathyroid function may be suppressed by dietary calcium in some patients with primary hyperparathyroidism.

Adult↗
PubMed1985

Messenger ribonucleic acid from tumors associated with humoral hypercalcemia of malignancy directs the synthesis of a secretory parathyroid hormone-like peptide.

Previous studies have provided evidence that tumors associated with humoral hypercalcemia of malignancy produce a factor that shares certain biological properties with, but is not, native PTH. In the present study, media from Xenopus oocytes microinjected with polyadenylated RNA prepared from three human or animal tumors associated with humoral hypercalcemia of malignancy are shown to have activity in the cytochemical bioassay for PTH. This activity parallels that of the PTH standard in the assay and is completely inhibited by the PTH analog [Nle8,Nle18,Tyr34]bovine PTH-(3-34)NH2. Media from oocytes injected with mRNA from control tumors contain no such activity. The mRNA encoding this activity in one of the animal tumors has been enriched approximately 10-fold by preparative polyacrylamide gel electrophoresis. These results demonstrate that tumors associated with humoral hypercalcemia of malignancy have the capacity for directing the synthesis of a secretory protein that fulfills certain criteria for being the humoral mediator in question. These techniques may provide the basis for molecular cloning of this factor.

Animals↗
PubMed1985

Elevated production rate of 1,25-dihydroxyvitamin D in patients with absorptive hypercalciuria.

Previous studies of 1,25-dihydroxyvitamin D [1,25-(OH)2D] kinetics in normal subjects using the pulse injection technique have led to conflicting results, and only limited data are available concerning 1,25-(OH)2D kinetics in hypercalciuric patients. We developed an infusion equilibrium technique that measures the metabolic clearance and production rates of 1,25-(OH)2D and applied this technique in 13 normal subjects and 9 well characterized patients with absorptive hypercalciuria; all subjects were studied after 10 days on a 400-mg calcium intake. All subjects received a constant infusion of [3H]1,25-(OH)2D3 (20,000 dpm/min). Purified plasma radioactivity reached steady state levels after 15 h, and between 15 and 19 h, serial measurements of purified plasma radioactivity and endogenous 1,25-(OH)2D were made for calculation of metabolic clearance and production rates. In the 13 normal subjects, the MCR values were within a narrow range, with a mean +/- SD value of 37 +/- 6 ml/min, which, when combined with the mean steady state concentration of endogenous 1,25-(OH)2D (42 +/- 6 pg/ml), yielded a mean production rate of 2.2 +/- 0.5 micrograms/day. In the 9 patients with absorptive hypercalciuria, MCR values also were tightly clustered, with a mean of 35 +/- 4 ml/min. However, the mean endogenous steady state 1,25-(OH)2D level was significantly elevated in these patients, such that the calculated mean 1,25-(OH)2D production rate was significantly elevated at 3.4 +/- 0.5 micrograms/day. In 7 of the 9 patients with absorptive hypercalciuria, production rates exceeded the highest values found in the normal subjects. These data demonstrate disordered 1,25-(OH)2D production as opposed to metabolic clearance in the syndrome of absorptive hypercalciuria.

Adult↗
PubMed1984

Evidence for disordered control of 1,25-dihydroxyvitamin D production in absorptive hypercalciuria.

In previous studies, we observed increases in the circulating concentration and production rate of 1,25-dihydroxyvitamin D (1,25-(OH)2D) in a large majority of patients with the syndrome of absorptive hypercalciuria. In the present study, the hypothesis that 1,25-(OD)2D production might be relatively autonomous in this syndrome was tested by fashioning a suppression test in which patients were challenged with a short-term increase in dietary calcium intake. We found that contrary to our hypothesis, the circulating concentration of 1,25-(OH)2D was remarkably sensitive to calcium intake in 15 patients with absorptive hypercalciuria (mean decrease, from 74 to 49 pg per milliliter, P less than 0.001). When this challenge was prolonged for two weeks, however, patients with absorptive hypercalciuria had evidence of an apparent "escape" phenomenon, in which the circulating concentration of 1,25-(OH)2D rebounded toward its initial level and the renal tubular phosphate threshold fell markedly. These findings provide evidence for disordered control of renal phosphate handling and 1,25-(OH)2D production in absorptive hypercalciuria and suggest a linked rather than a cause-and-effect relation between these two abnormalities.

Adult↗
PubMed1984

Oxalate absorption and postprandial urine supersaturation in an experimental human model of absorptive hypercalciuria.

The effect of 1.25-dihydroxyvitamin D [1,25-(OH)2D] on dietary oxalate absorption and postprandial urine supersaturation with calcium oxalate was determined in 11 normal subjects. 1,25-(OH)2D increased the urinary excretion of orally administered [14C]oxalate in the 8 h period after a liquid meal containing 1.875 mmol of calcium and 0.83 mmol of oxalate (P less than 0.01), and during a 48 h period when the subjects ingested a diet containing 25 mmol of calcium and 3.3 mmol of oxalate/day (P less than 0.01); however, 1,25-(OH)2D administration had no effect on [14C]oxalate excretion when calcium was removed from the liquid meal. 1.25-(OH)2D increased 24 h urinary oxalate excretion from 28.7 +/- 2.1 mmol/mol of creatinine to 36.8 +/- 2.6 mmol/mol of creatinine (P less than 0.05) on the 10 mmol/day calcium diet and from 26.4 +/- 2.9 to 33.2 +/- 2.2 mmol/mol of creatinine (P less than 0.1) on the 25 mmol/day calcium diet. A linear correlation (r = 0.72) was found between plasma 1,25-(OH)2D levels and urinary [14C]oxalate excretion after the liquid meal. 1,25-(OH)2D administration produced postprandial supersaturation of urine with calcium oxalate and calcium oxalate crystalluria. These studies suggest that 1,25-(OH)2D increases oxalate absorption (and urinary excretion) by increasing calcium absorption, which results in less binding of calcium to oxalate in the intestine; therefore more oxalate is available for absorption. The combined effect of increased calcium and oxalate absorption results in postprandial supersaturation of urine with calcium oxalate, with resultant crystalluria.

Adult↗
PubMed1984

Biochemical and histomorphometric characterization of a rat model for humoral hypercalcemia of malignancy.

Humoral hypercalcemia of malignancy is a common but incompletely understood syndrome in humans. In an effort to define an animal model of this syndrome, we studied the transplantable Rice-500 Leydig cell tumor in male Fisher rats. Animals were studied 4-9, 10-12, and 13-14 days after tumor transplantation. By days 13-14, tumor-bearing animals were significantly hypercalcemic, hypercalciuric, and hyperphosphaturic compared to control animals. Fractional phosphorus excretion was elevated 4-fold in the tumor-bearing group despite hypophosphatemia. Mean nephrogenous cAMP in the tumor-bearing animals was 5 times the value in controls at days 13-14, while simultaneous immunoreactive PTH levels were undetectable. Plasma 1,25-dihydroxyvitamin D was significantly elevated in the tumor-bearing animals on day 14. Quantitative bone histomorphometry showed uncoupling of bone cell function in the tumor group, with marked suppression of bone formation, while indices of bone resorption were more than 2-fold elevated. Conditioned medium from tumor cells grown in culture consistently showed activity in a fetal bone-resorbing assay. This activity was heat stable and had an estimated mol wt of 30,000-50,000 daltons. Incubation of cells with indomethacin had no effect on bone-resorbing activity. These data indicate that the mediator in the model shares some of the actions of PTH, but is clearly distinct from native PTH. The findings exactly parallel those in human humoral hypercalcemia of malignancy, with the elevated 1,25-dihydroxyvitamin D values being the sole exception. The demonstration of in vitro bone-resorbing activity will aid in further characterization of the mediator.

Animals↗
PubMed1984

A consideration of the hormonal basis and phosphate leak hypothesis of absorptive hypercalciuria.

Fifty patients with absorptive hypercalciuria (AH), 25 normal subjects (NS), and 25 nonhypercalciuric patients with stone disease (NHSF) were studied using an oral calcium tolerance test and 24-h urine collections on both a restricted and an unrestricted calcium intake. Mean (+/- SD) fasting fractional calcium excretion was increased in the patients with AH (2.7 +/- 1.1% vs. 1.4 +/- 0.6% in the NS; P less than 0.001) and was negatively correlated with fasting nephrogenous cAMP, suggesting that this renal calcium leak was secondary to parathyroid suppression. Plasma 1,25-dihydroxyvitamin D [1,25-(OH)2D] was elevated in 80% of patients with AH and was high normal in the remaining 20%. Ten patients, selected on the basis of results for 1,25-(OH)2D greater than 4 SD from the normal mean, displayed a particularly severe pattern of abnormalities, including mild hypercalcemia in two patients. Pooled data from the NS and patients with AH revealed a significant negative correlation between the plasma concentration of 1,25-(OH)2D and the renal phosphate threshold (r = -0.40; P less than 0.001), but this correlation lost significance when the NHSF were substituted for the NS as a control group (r = -0.07; P = NS). These findings 1) provide a pathophysiological basis for the increase in fasting calcium excretion commonly observed in hypercalciuric patients, and 2) stress the importance of circulating 1,25-(OH)2D in the pathogenesis of the syndrome, but 3) fail to support the phosphate leak theory of pathogenesis.

Adult↗
PubMed1984

An experimental human model of 1,25-dihydroxyvitamin D-mediated hypercalciuria.

An increase in fasting calcium excretion occurs in hypercalciuric patients and has been interpreted by many investigators as evidence for a primary renal tubular leak of calcium. In a recent series of 50 patients with absorptive hypercalciuria, we found a mean increase in fasting fractional calcium excretion (calcium clearance) and provided data suggesting that this leak of calcium was secondary to intestinal hyperabsorption of calcium and suppression of parathyroid secretion. To examine this question, a model of 1,25-dihydroxyvitamin D [1,25-(OH)2D]-mediated hypercalciuria was created by administering a large dose of 1,25-(OH)2D to normal subjects. In addition to the expected features of absorptive hypercalciuria during 1,25-(OH)2D administration, the subjects had an increase in fasting calcium excretion and a marked increase in calcium excretion during a restricted calcium diet, points that might be interpreted as favoring a resorptive and/or renal component to the net hypercalciuria. However, total hydroxyproline excretion remained unchanged (18.0 vs. 19.0 mg/g creatinine), and the increase in fasting calcium excretion was found to reflect an increase in calcium clearance; the latter was inversely correlated with parathyroid function, as determined by fasting measurements of nephrogenous cAMP excretion (r = -0.77; P less than 0.01). We conclude that measurements of calcium excretion in the fasting state or on a restricted calcium diet do not represent valid criteria for differential diagnosis of the hypercalciurias.

Adult↗