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Available androgen binding to soluble proteins from the cytosol of human endometrium was studied using the dextran coated charcoal adsorption method and sucrose density centrifugation analysis. Specific binding of [3H]-5 alpha-dihydrotestosterone ([3H]-DHT) was observed with both methods. The apparent dissociation constant (Kd), for DHT binding is 1.3 +/- 0.2 (SEM) nM and the binding capacity 177 +/- 42 (SEM) fmol/mg protein. Sucrose density ultracentrifugation identifies specific [3H]-DHT binding that sediments at 4S and 8S. The stability of the androgen receptor in human endometrium is increased by the addition of 10% glycerol to the homogenization buffer. The addition of trypsin or pronase and heating at 60 degrees C reduces specific binding which demonstrates that the specific [3H]-DHT binder is a protein. The uptake of [3H] DHT in endometrial tissue minces indicated that 20% of the bound radioactivity was nuclear. Steroid specificity suggests that the binding protein from the uterus is specific for androgens. These observations indicate that androgen binding protein in the human uterus has the characteristics of the androgen receptor.
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PMID:The androgen receptor of the human endometrium. 358 78

The effect of peptide YY (PYY) on gastric and pancreatico-biliary secretion was studied in humans. Peptide YY was infused into groups of 6 healthy volunteers at doses of 0.59, 0.20, and 0.064 pmol X kg-1 X min-1. The two higher doses caused a significant suppression of gastric acid and pepsin output during background stimulation with pentagastrin. The middle dose of PYY (0.20 pmol X kg-1 X min-1) that increased plasma PYY levels by 27 +/- 2 pM caused a 90% +/- 18% (mean +/- SEM; p less than 0.001) reduction in the incremental gastric volume response to pentagastrin. Similarly this dose of PYY caused a substantial inhibition of the acid (77% +/- 14%; p less than 0.005) and pepsin (96% +/- 22%; p less than 0.01) response to pentagastrin; in 2 subjects, pepsin output fell to below basal levels. In contrast, the highest dose of PYY (0.62 pmol X kg-1 X min-1) had no significant influence on duodenal juice volume, output of bicarbonate, trypsin, or bilirubin during low dose stimulation with secretin (0.25 pmol X kg-1 X min-1) and cholecystokinin-8 (0.15 pmol X kg-1 X min-1). Thus PYY concentrations in the circulation similar to those seen after the ingestion of food cause a marked reduction in gastric secretion. This peptide should therefore be considered as one of the possible candidates for the classical enterogastrone.
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PMID:Effect of peptide YY on gastric, pancreatic, and biliary function in humans. 383 79

Antibodies have been raised against biosynthetic human proinsulin that show less than 1% cross-reactivity with human insulin and C-peptide. A sensitive (IC50 0.16 pmol/ml; minimum detectable concentration 0.004 pmol/ml) radioimmunoassay has been developed using this antiserum and 125I-proinsulin that will measure proinsulin-like immunoreactivity in human serum without the need for prior separation of insulin or C-peptide. In healthy, fasted subjects (N = 23), the serum proinsulin concentration was 0.015 +/- 0.001 pmol/ml (mean +/- SEM). In six healthy subjects, serum proinsulin rose to 250% of basal after 120 min in response to 100 g oral carbohydrate, but to only 130% after 60 min following 25 g oral carbohydrate. The proinsulin/total immunoreactive insulin ratio and the proinsulin/C-peptide ratio fell sharply after both high and low carbohydrate loads. Endogenous human serum proinsulin-like immunoreactivity released into the circulation after 100 g carbohydrate was eluted from a Mono Q high-performance, ion-exchange column with the same retention time as biosynthetic human proinsulin. Treatment of biosynthetic proinsulin with trypsin under mild conditions led to a decrease in proinsulin-like immunoreactivity concomitant with an increase in C-peptide and insulin-like immunoreactivity, indicating that the proinsulin-specific antiserum did not preferentially recognize intermediates of proinsulin cleavage.
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PMID:Measurement of circulating human proinsulin concentrations using a proinsulin-specific antiserum. 388 62

The rate of tissue kallikrein (EC 3.4.21.35) excretion into the urine has been examined with an active site-specific radioimmunoassay for kallikrein in renal transplant recipients, in post-uninephrectomy kidney donors, and in a normal control population. Normal individuals on uncontrolled diets excreted 96.88 +/- 7.00 (SEM) micrograms of active kallikrein/24 hr and 113.68 +/- 8.39 micrograms of total kallikrein/24 hr, as determined after trypsin treatment of urine samples. Uninephrectomized donors secreted significantly less (P less than 0.05) active (44.99 +/- 6.39 micrograms/24 hr) and total (73.59 +/- 11.95 micrograms/24 hr) kallikrein than either the entire normal population or an age-matched subpopulation. Recipients with good renal function who had received kidneys 2 to 13 years prior to kallikrein assay excreted less (P less than 0.05) active (13.21 +/- 2.50 micrograms/24 hr) and total (18.69 +/- 3.65 micrograms/24 hr) kallikrein than either normal or uninephrectomized populations. Similar values for active (11.05 +/- 1.56 micrograms/24 hr) and total (17.60 +/- 1.96 micrograms/24 hr) kallikrein were seen in patients who had received kidneys within 6 months of assay. Thus, kallikrein excretion in kidney recipients remains significantly lower than in uninephrectomized donors. As compared to normal individuals, the reduced kallikrein excretion in post-uninephrectomized kidney donors and in renal allograft recipients suggests that renal kallikrein excretion may reflect functional distal tubular mass.
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PMID:Kallikrein excretion in renal transplant recipients and in uninephrectomized donors. 390 May 31

The authors wished to determine whether secretory cell metaplasia (SCM) induced in the bronchi of hamsters by human neutrophil elastase (HNE) was enzymatically mediated. We also wished to determine whether SCM could be induced by a proteolytic enzyme devoid of elastolytic activity. Accordingly, groups of weight-matched hamsters were given a single intratracheal instillation of 0.5 ml of saline solution containing one of the following: 300 micrograms of HNE purified from blood neutrophils, n = 14; 300 micrograms of HNE inactivated with Suc-Ala-Ala-Pro-Val chloromethyl ketone (CMK), n = 10; 500 micrograms of porcine pancreatic trypsin treated with CMK to eliminate residual active elastase, n = 10; 500 micrograms of trypsin inactivated by tosyl lysine chloromethyl ketone, n = 10; 2 micrograms CMK, n = 10; and saline alone, n = 10. Seven untreated animals served as additional controls. Twenty-one days post treatment, 5-6 micron paraffin-embedded sections, from the left lung hilar region, stained by Alcian blue and periodic acid-Schiff reaction were graded on a five-point scale for determination of the secretory cell index, which reflects SCM. Both elastase and trypsin produced severe SCM: mean +/- SEM secretory cell indices were 2.96 +/- 0.11 and 2.72 +/- 0.19, respectively, compared with values of 0.90 +/- 0.35 for the untreated group and 0.93 +/- 0.46 for the saline group (p less than .05). The secretory cell indices of the groups treated with inactivated elastase or trypsin were comparable to those of the saline-treated and untreated groups.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Proteolytic activity of human neutrophil elastase and porcine pancreatic trypsin causes bronchial secretory cell metaplasia in hamsters. 390 65

Using three antisera to oxytocin (OT Pitt Ab-1, OT Pitt Ab-2, and TOR OT Ab), we found comparable levels of OT in response to infant suckling and during infusion of synthetic OT, and identical standard curves with biological and synthetic standards of OT. Pitt Ab-1, but not Pitt Ab-2 or TOR OT Ab, measured increased OT in response to estrogen. Using an arginine vasotocin RIA (TOR AVT Ab), we found an increase in AVT immunoreactivity after estrogen treatment. Mean basal OT levels measured with OT Pitt Ab-2 in plasma of men [0.75 +/- 0.06 (+/- SEM) microU/ml] and women (0.8 +/- 0.09 microU/ml) were lower than OT measured with Pitt Ab-1 (1.7 +/- 0.09 microU/ml in men and 1.7 +/- 0.07 microU/al in women; P less than 0.001). Mean OT measured with Pitt Ab-2 in the plasma of women given estrogen chronically (0.8 +/- 0.04 microU/ml) and acutely (0.6 +/- 0.15 microU/ml) were not significantly different from basal levels. OT levels measured with Pitt Ab-1 in the same samples were 4.6 +/- 0.5 and 4.3 +/- 0.5 microU/ml, respectively, both significantly increased from basal levels (P less than 0.001) and significantly higher than OT measured with Pitt Ab-2 (P less than 0.001). Mean OT measured with Pitt Ab-1 in the plasma of pregnant women was 8.6 +/- 1.02 microU/ml, significantly higher than OT measured with Pitt Ab-2 (1.0 +/- 0.3 microU/ml; P less than 0.001). Men given 25 mg diethylstilbestrol had significant increases in OT measured with Pitt Ab-1 and in AVT measured with TOR AVT (P less than 0.01), but not in OT measured with Pitt Ab-2. Plasma from a man given diethylstilbestrol was prepared for high performance liquid chromatography and applied to a C18 muBondapak reverse phase column. The plasma contained two peaks of immunoreactivity detected as OT with Pitt Ab-1 and as AVT using TOR AVT Ab. The material was not detected by Pitt Ab-2 or TOR OT Ab and did not coelute with standards of OT, AVT, or AVP. Pregnancy plasma, thioglycolic acid, chymotrypsin, and trypsin reduced Pitt Ab-1, Pitt Ab-2, and TOR OT immunoreactivity of synthetic OT. The percent recovery of OT immunoreactivity was not significantly different with Pitt Ab-1 vs. Pitt Ab-2. A novel peptide, which is increased in response to administered estrogen, is present in human plasma and is detected by some antisera to OT and AVT. The observation explains the wide variability in OT levels in the estrogen-primed state and provides a new mechanism to study estrogen-related physiology and pathophysiology.
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PMID:A novel oxytocin-like and vasotocin-like peptide in human plasma after administration of estrogen. 396 93

The degree of tyrosine sulfation and the distribution between gastrin-17- and gastrin-34-like immunoreactivity (LI) were studied in the antra of ten mammalian species. Specific radioimmunoassays, gel-, and ion-exchange chromatography as well as enzymatic cleavage with trypsin and arylsulfatase were used. The percentage of sulfation varied from 24.4 +/- 4.2 (mean +/- SEM) in dogs to 80.1 +/- 2.6 in sheep, 46.8 +/- 3.3 in humans, 50.1 +/- 3.2 in cows, 55.9 +/- 2.3 in rats, 57.4 +/- 3.1 in pigs, 61.3 +/- 2.2 in guinea pigs, 64.1 +/- 4.7 in cats, 64.8 +/- 2.1 in mice and 68.2 +/- 2.8 in rabbits. Gastrin-34-LI in antral extracts could be converted to gastrin-17-LI by trypsin in all species. Five percent of antral gastrins eluted as gastrin-34-LI in all species. We conclude that while the ratio of gastrin-34-LI to gastrin-17-LI varies little in mammals, large differences occur in the degree of sulfation.
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PMID:Species variation in the tyrosine sulfation of mammalian gastrins. 398 36

Sera from 15 patients with the Zollinger-Ellison syndrome were subjected to gel filtration on Sephadex G-50 superfine columns (10 x 2000 mm). The concentration of gastrin in the effluent was determined by a sensitive radioimmunoassay. Immunoreactive gastrin was eluted in four components in 14 sera. (1) Component I, eluted in the same position as proinsulin, constituted 9.7 +/- 1.2 (mean +/- SEM)% of the total immunoreactivity. (2) Component II (;big gastrin') eluted between proinsulin and insulin constituted 57.8 +/- 4.1% (mean +/- SEM) of immunoreactive gastrin. In three sera with the highest concentration of gastrin, component II appeared biphasic. (3) Component III (;little gastrin') was distributed in two peaks; the first one eluted in the same position as the heptadecapeptide gastrin II made up 17.4 +/- 2.7 (mean +/- SEM)% of the total immunoreactivity; the second one eluted in the same position as gastrin I constituted 9.5 +/- 1.3 (mean +/- SEM)%. (4) Component IV (;minigastrin') was eluted immediately before the salt peak and constituted 5.6 +/- 1.4 (mean +/- SEM)%. In one serum only components I and II were present. After incubation with trypsin all immunoreactivity in components I and II was converted to heptadecapeptide-like gastrins.The findings suggest that immunoreactive gastrin in serum from Zollinger-Ellison patients is circulating in at least four components of different molecular size.
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PMID:Gel filtration studies on immunoreactive gastrin in serum from Zollinger-Ellison patients. 419 48

Rat renal lymph contains 254 +/- 17 ng/ml (means +/- SEM, N = 20) of immunoreactive glandular kallikrein. Like the immunoreactive glandular kallikrein in plasma, it is biologically inactive. Gel filtration of renal lymph reveals profiles for immunoreactive glandular kallikrein, protein, and inhibition of trypsin and kallikrein which resemble those seen for plasma except that high molecular weight plasma components are reduced or missing in renal lymph. In contrast, gel filtration of thoracic lymph reveals immunoreactive glandular kallikrein and protein profiles which are indistinguishable from those seen with plasma. Renin levels are 170-fold higher in renal lymph than in thoracic lymph while angiotensin-converting enzyme levels are only 16% those of thoracic lymph. In keeping with the high renin and low converting enzyme activities, renal lymph contains high levels of angiotensin I. Immunoreactive glandular kallikrein levels in renal lymph, thoracic lymph and plasma do not show the striking differences observed for renin.
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PMID:Kallikrein-kinin and renin-angiotensin systems in rat renal lymph. 608 86

A protein fraction (60-F), obtained from cell-free extract of living hemolytic streptococcus, Su-strain, by 50-60% saturation with ammonium sulfate, inhibited the de novo synthesis of nucleic acids and proteins in Ehrlich ascites carcinoma (EAC) cells. 60-F released RNA but not DNA from EAC cells. This cytotoxic or cytolytic effect of 60-F was substantiated morphologically by scanning and transmission electron microscopic (SEM and TEM) observations, showing that 60-F induced cellular changes such as a loss of microvilli, bleb formation, cell deformity and partial gap of cell membrane in EAC cells. In addition, it was found that 60-F was more stable in RPMI-1640 medium supplemented with fetal calf serum than in other various media examined and was sensitive to temperature, pH changes and trypsin digestion.
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PMID:Biochemical and electron microscopic observations of cytotoxic effect of a fraction from hemolytic streptococcus on Ehrlich ascites carcinoma cells. 618 69


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