Excretion of progastrin products in human urine.

The renal handling of carboxyamidated gastrins, NH2-terminal progastrin fragments, and glycine-extended gastrins was examined in healthy volunteers. The respective urinary clearances after a meal amounted to 0.09 +/- 0.02%, 0.17 +/- 0.04% (P < 0.05), and 0.04 +/- 0.01% (P < 0.01) of the glomerular filtration rate. During intravenous infusion of carboxyamidated gastrin-17, progastrin fragment-(1-35), and glycine-extended gastrin-17, the respective urinary clearances amounted to 0.08 +/- 0.02, 0.46 +/- 0.08, and 0. 02 +/- 0.01%, respectively, of the glomerular filtration rate. The metabolic clearance rate of the three peptides was 24.4 +/- 1.3, 6.0 +/- 0.4, and 8.6 +/- 0.7 ml. kg-1. min-1. A maximum rate for tubular transport or degradation of the peptides could not be determined, nor was a renal plasma threshold recorded. Plasma concentrations and urinary excretion rates correlated for gastrin-17 and progastrin fragment-(1-35) (r = 0.94 and 0.97, P < 0.001), whereas the excretion of glycine-extended gastrin diminished with increasing plasma concentrations. We conclude that renal excretion of progastrin products is negligible compared with renal metabolism and that renal handling of the peptides depends on their molecular structure. Hence, the kidneys exhibited a higher excretion of NH2-terminal progastrin fragments than of carboxyamidated and especially glycine-extended gastrins.

Renal tubular transport and metabolism of carboxyamidated and glycine-extended gastrins in pigs.

Renal handling of postprandial and intravenously administered gastrin was investigated in anaesthetised pigs. The fractional extraction of postprandial carboxyamidated and glycine-extended gastrin in the kidneys was 0.21 +/- 0.01 and 0.16 +/- 0.02, but the respective urinary clearance comprised only 0.57 +/- 0.03 and 0.44 +/- 0.05% of the GFR (P < 0.02). The respective total body clearance of carboxyamidated and glycine-extended gastrin-17 (gastrin-17 and gastrin-17Gly) during continuous infusion was 22.9 +/- 1.5 and 19.6 +/- 1.4 mL kg-1 min-1 (NS), and the renal fractional extraction of the peptides was 0.31 +/- 0.03 and 0.29 +/- 0.05, respectively. The kidneys accounted for 8% of total body clearance of gastrin-17. Renal filtration rate of gastrin-17 exceeded renal extraction rate (9.739 +/- 0.487 vs. 6.407 +/- 0.321 pmol min-1). Urinary clearance of gastrin-17 and gastrin-17Gly amounted only 0.91 +/- 0.16 and 0.13 +/- 0.03%, respectively, of the GFR (P < 0.01), but urinary excretion rate correlated with the filtered amount of the peptides (r = 0.93, P < 0.01). Neither was a renal plasma threshold recorded nor was a Tm value for tubular uptake or degradation of gastrin achieved in spite of supraphysiological plasma levels of the peptides. The results indicate that filtered gastrin is almost completely removed in the renal tubules, primary by metabolism although part of the absorbed peptides may be returned to the circulation in intact form. The process for uptake or metabolism has a high capacity but varies with the molecular form of gastrin.

Mobilization of gastric histamine during repeated administration of a proton potassium adenosine triphosphatase inhibitor in intact and antrectomized rats.

Intact and antrectomized female rats were treated with the potent proton pump inhibitor, E3810 (daily 40 mg/kg weight, s.c.) for 4 weeks. Plasma gastrin concentration and urinary excretion of N-terminal big gastrin increased until day 14 and persisted at a high level in intact rats treated with E3810, but did not increase in antrectomized rats. Urinary excretion of histamine increased progressively and reached 7 times the control value following 4 weeks of treatment with E3810 in intact rats, but not in antrectomized rats. At the termination of the treatment, the endocrine cell density in the oxyntic mucosa of intact rats had increased by 85% with increased histamine content and elevated histidine decarboxylase activity, while antrectomized rats showed a low histamine level and low histidine decarboxylase activity. Administration of gastrin-17 I (10 micrograms/kg weight, sc) itself caused a significant increase in urinary excretion of histamine, which was inhibited by the specific gastrin receptor antagonist, L-365,260. These results suggests that the massive urinary excretion of histamine caused by the treatment with E3810 reflects gastrin-induced mobilization of gastric histamine and that neither E3810 itself nor E3810-induced luminal pH elevation has direct effects on mobilization of oxyntic mucosal histamine.

Urinary gastrin output and serum gastrin in patients with liver cirrhosis. Urinary gastrin output in cirrhosis.

The aim of the present study was to examine the diurnal urinary gastrin output in cirrhotics and to clarify whether in patients with hepatorenal syndrome urinary gastrin output is reduced. Thirty-two cirrhotics and 25 age- and sex-matched, controls were studied. Cirrhotics were divided into 3 groups: (I: without ascites and normal serum creatinine; II: ascites and normal creatinine; III: ascites and increased creatinine). Mean fasting serum gastrin concentration was lower in the control group than in Group I, II (p < 0.01) or III (p < 0.001). In this latter group mean serum gastrin concentration was significantly higher (p < 0.001) than in the other two groups of cirrhotics. The mean 24 h urinary gastrin output was lower (p < 0.001) in Group III patients than in the other groups of subjects studied. Also in the controls urinary gastrin output was lower (p < 0.01) than in Groups I and II. These findings suggest that: a) in cirrhotics with normal serum creatinine the average serum gastrin levels over the course of the day are indeed higher than in normals and b) In cirrhotics with hepatorenal syndrome, impaired urinary gastrin output appears to contribute significantly to their hypergastrinemia.

Effect of antrectomy and drug-induced achlorhydria on urinary excretion of N-terminal big gastrin immunoreactivity in rats.

Immunoreactivities of urinary N-terminal big gastrin and serum C-terminal gastrin were determined in intact and antrectomized rats by radioimmunoassay using two antisera specific for N- and C-termini of big gastrin, respectively. Gel filtration of urine extract from intact rat showed a single giant peak of N-terminal big gastrin immunoreactivity eluted in a later position than 1-17 gastrin-34, indicating that N-terminal peptides smaller than 1-17 gastrin-34 are excreted in urine. Serum C-terminal gastrin concentration in antrectomized rats was about one sixth that in intact rats. Urinary excretion of N-terminal big gastrin in antrectomized rats was about one sixth that in intact rats. 2 week treatment with E3810, a proton pump inhibitor, (40 mg/kg/day, s.c.) induced urinary excretion of N-terminal big gastrin in parallel with a marked increase in serum C-terminal gastrin concentration in intact rats. Antrectomy completely prevented both the increase in urinary excretion of N-terminal big gastrin and the elevation of serum C-terminal gastrin induced by administration of E3810. There was an excellent correlation between serum concentration of C-terminal gastrin and urinary excretion of N-terminal big gastrin. These results suggest that urinary N-terminal big gastrin, which mostly originates from the gastric antrum, is a useful indicator of gastrin secretion in the rat.

Plasma concentrations and urinary excretion of regulatory peptides in patients with urticaria pigmentosa.

Patients with urticaria pigmentosa were investigated during symptom-free interval regarding plasma concentrations and urinary excretion of immunoreactive regulatory peptides: calcitonin gene-related peptide (CGRP), gastrin, neurokinin A (NKA), neuropeptide Y (NPY), somatostatin (SOM), substance P (SP) and vasoactive intestinal peptide (VIP). The plasma concentrations of these peptides, except for CGRP, were below the detection limit. The urinary excretion of the regulatory peptides were not higher in the patient group than in the controls, but in individual patients there was high urinary excretion of SP and VIP. A lower urinary excretion of CGRP was found in the patient group in addition to a tendency to a lower plasma concentration.

Purification of N-terminal hexapeptide of big gastrin from human urine.

We previously demonstrated that extremely high amounts of N-terminal big gastrin (G-34) fragments are excreted in human urine and three of them are N-terminal octa-, nona-, and decapeptide of G-34. Our subsequent examination revealed that there exists a considerable amount of another N-terminal G-34 fragment in urine, less hydrophobic than the three peptides. We purified this fragment from urine of an achlorhydric patient and determined the structure: less than Glu-Leu-Gly-Pro-Gln-Gly. The purification was carried out by Sep-Pak C18 cartridges, Sephadex G-25, and reverse phase HPLC. The structure was determined by a combination of amino acid analysis, amino acid sequence analysis, and mass spectral analysis. N-terminal hexapeptide of G-34 is the second richest component of urinary N-terminal G-34 fragments next to N-terminal octapeptide of G-34 in normal subjects.

Temporal profiles of urinary excretion of NH2-terminal big gastrin immunoreactivity in humans.

We studied the temporal profile of urinary NH2-terminal big gastrin immunoreactivity (NT G-34-IR) excretion in order to evaluate the dynamics of gastrin secretion. The temporal profile of urinary NT G-34-IR excretion in normal subjects represented three peaks corresponding to each meal. In contrast, the profile in antrectomized patients and patients under total parenteral nutrition (TPN) represented a flat pattern. Urinary NT G-34-IR excretions during fasting 2-h periods in antrectomized patients and TPN patients were about one-sixth and one-third, respectively, of basal NT G-34-IR excretion in normal subjects (53.1 +/- 13.9 pmol/h). Total urinary NT G-34-IR excretion during 24 h both in antrectomized patients (220 +/- 35 pmol/24 h) and TPN patients (390 +/- 68 pmol/24 h) was also significantly lower than in normal subjects (1985 +/- 403 pmol/24 h). The present study showed that the main source of urinary NT G-34-IR is the gastric antrum, that the main factor fluctuating its excretion is food intake, and that long-term TPN reduces basal gastrin secretion. Urinary NT G-34-IR would be a useful indicator for total gastrin secretion.

A possible indicator of urinary NH2-terminal big gastrin immunoreactivity for gastrin secretion.

Urinary and plasma gastrin immunoreactivities in normal subjects were studied by radioimmunoassays using three region-specific antisera. Urinary excretion of NH2-terminal big gastrin immunoreactivity (NT G-34-IR) in the fasting state (0.79 +/- 0.17 pmol/kg/h, mean +/- SE) was several hundred times as much as that of either COOH-terminal gastrin or gastrin/cholecystokinin immunoreactivity. Urinary NT G-34-IR increased significantly after feeding, and correlated closely with integrated plasma NT G-34-IR. Renal clearance of NT G-34-IR was 62.4 +/- 7.9 ml/min and about one hundred times greater than that of any other gastrin immunoreactivities, indicating that there exist different catabolic pathways of gastrin peptides in the kidney. Gel filtration of urine extract revealed that a single giant peak of NT G-34-IR eluted in a later position than NH2-terminal heptadecapeptide of big gastrin. These results suggest that most of plasma NT G-34-IR is excreted in urine, and urinary excretion of NT G-34-IR reflects well-integrated plasma NT G-34-IR. Therefore, urinary NT G-34-IR may serve as a feasible indicator for gastrin secretion.

NH2-terminal big gastrin immunoreactivity in human urine.

The concentrations and molecular forms of urinary and plasma gastrin from normal subjects were studied by radioimmunoassays using two region-specific antisera. Urinary concentration of NH2-terminal big gastrin (G-34) immunoreactivity was several hundred times as great as that of COOH-terminal gastrin immunoreactivity. Fractionation of urine extract showed a broad giant peak of NH2-terminal G-34 immunoreactivity (gastrin fragments "U") eluting in a later position than G-34(1-17) by Sephadex G-50 column chromatography. HPLC revealed that urinary NH2-terminal G-34 immunoreactivity was composed of four fragments including G-34(1-8), G-34(1-9), and G-34(1-10). Sephadex G-50 column chromatography of plasma extract revealed two or three peaks of NH2-terminal G-34 immunoreactivity, and a major peak eluted in the same position as urinary gastrin fragments "U". These results and data on renal clearances suggest that most of all gastrin fragments "U" in plasma are excreted in urine without renal reabsorption, whereas almost all of plasma COOH-terminal gastrin peptides including G-34 and little gastrin (G-17) are removed and metabolized in the kidney.

Purification and structural determination of urinary NH2-terminal big gastrin fragments.

We previously demonstrated that there existed extremely abundant NH2-terminal big gastrin immunoreactivity (NT G-34-IR) in human urine. This report describes the purification and sequence of NT G-34-IR from the urine of an achlorhydric patient. The purification was carried out by a combination of Sep-Pak C18 cartridges, Sephadex G-25, and HPLC steps using a radioimmunoassay specific for NH2-terminus of G-34 and ultraviolet absorption at 214 nm as monitors. Three peptides were isolated. The amino acid analysis, mass spectrometry, and sequence analysis confirmed the structures of urinary NT G-34 fragments being less than Glu-Leu-Gly-Pro-Gln-Gly-Pro-Pro, less than Glu-Leu-Gly-Pro-Gln-Gly- Pro-Pro-His, and less than Glu- Leu-Gly-Pro-Gln-Gly-Pro-Pro-His-Leu. NH2-terminal octapeptide of G-34 was the main component of urinary NT G-34-IR.

Distribution, molecular forms, and bioactivity of immunoreactive gastrin in a patient with metastatic gastrinoma.

A patient with metastatic gastrinoma whose plasma gastrin ranged from 7.6 nmol X ml-1 at presentation to 18.9 nmol X ml-1 just before death 4 yr later has been studied. The presence of such high circulating levels facilitated study of the biological and immunological activities as well as the molecular forms of gastrin in plasma, ascitic fluid, urine and cerebrospinal fluid. The results indicate: 1) the 34 amino acid gastrin (G34) was the only molecular form detectable in the body fluids studied; 2) bioactivity of G34 is maintained despite passage across the renal endothelium and the blood brain barrier; 3) the blood brain barrier significantly limits diffusion of G34.

Urinary immunoreactive gastrin in normal subjects.

Gastrin can readily be concentrated from 10 to 50 ml of urine with better than 90% recovery using octadecylsilyl (ODS) silica columns (C18 Sep-Pak cartridge) and then measured by radioimmunoassay. Fractionation on Sephadex G50 gel filtration reveals that the apparent immunoreactivity is not due to nonspecific interference in the assay system but does correspond to the two known forms of gastrin, the 17 and 34 amino acid peptides. Renal clearance of gastrin in 5 normal subjects does not appear to differ in the fasted and fed state and ranged from 0.09 to 0.26 ml/min with an average of 0.16 +/- 0.05 (S.D.) ml/min. Urinary gastrin excretion in the overnight fasting state was generally less than 0.005 pmol/hr/kg body weight and fell to lower levels after a 20-hour fast. Increased urinary gastrin output was observed following feeding. Gastrin output in urine in 7 subjects ranged from 6.8 to 10.2 pmol/24 hr with an average of 8.5 +/- 1.5 (S.D.) pmol/24 hr. A single determination of renal gastrin clearance and 24-hour gastrin urinary output appears to be sufficient for the determination of averaged plasma gastrin levels in normal subjects without renal disease. Similar methodology should be applicable to a variety of other peptidal hormones as well.

Immunochemical studies on gastrins in the urine.

The concentration and molecular form of gastrin in urine were determined radioimmunochemically. Urine from hypergastrinaemic patients (Zollinger-Ellison syndrome and pernicious anaemia) contained gastrins corresponding to the serum components I and II. The excretion of gastrin increased with increasing gastrin concentrations in serum. Urine from six subjects with normal concentrations of gastrin in serum contained "apparent" gastrin immunoreactivity which could not be removed by specific immunoabsorption. No gastrin was detectable by gel filtration of desalted and concentrated urine from normal subjects. The apparent immunoreactivity was due partly to interference by sodium chloride. The results indicate that hypergastrinaemic patients, in contrast to normal subjects, excrete gastrins in the urine.

Role of the kidney in gastrin metabolism.

Fasting blood gastrin level among various clinical entities was measured by means of radioimmunoassay. Hypergastrinemia of various degrees was found in some cases of chronic renal failure. During angiography arteriovenous difference of gastrin level was investigated in the kidney in 22 patients without renal diseases. The high rate of gastrin removal from the blood circulation by the kidney was also confirmed in the synthetic human gastrin 1-17 (SHG 1-17) infusion experiment in dogs. In this study, hypergastrinemia seen in renal diseases was thought to be caused partly by impaired gastrin metabolism in the kidney.