Prostaglandin D2 metabolite in urine is an index of food allergy.

Food allergy is immediate hypersensitive reactions to ingested foods. Since early diagnosis is effective for disease control, development of an objective diagnostic index is required. Using mediator-lipidomics, we found that levels of the urinary prostaglandin D2 (PGD2) metabolite, tetranor-PGDM, reflected the severity of the allergic symptoms and intestinal mast cell hyperplasia in mice. Repeated oral challenges with ovalbumin promoted allergic symptoms in sensitized mice. Particularly, the allergic mice presented with increased numbers of intestinal mast cells, which strongly expressed hematopoietic PGD synthase (H-PGDS). The levels of urinary tetranor-PGDM increased as the disease progressed. Treatment with a mast cell inactivator or an anti-inflammatory steroid attenuated these symptoms and decreased the tetranor-PGDM urinary levels. The levels of urinary tetranor-PGDM did not correlate with the disease severity in murine models of colitis, asthma, or allergic dermatitis. Furthermore, we have shown that urinary levels of tetranor-PGDM were significantly higher in patients with food allergy than those in healthy volunteers and patients with other types of allergic diseases such as asthma, allergic rhinitis, and atopic dermatitis. These findings suggest that urinary tetranor-PGDM is a useful diagnostic index of food allergy in both mice and humans.

Urinary protease inhibitor Serpin B3 is higher in women and is further increased in female patients affected by aldosterone producing adenoma.

A correct diagnosis of primary aldosteronism (PA) requires adrenal venous sampling (AVS) for the classification of subtypes (bilateral hyperplasia, BAH, or adenoma, APA). Since such testing is not easily practicable, appropriate markers for the definition of subtypes are desirable. We hypothesized that an aldosterone excess was associated with abnormalities in urinary proteome, specific for PA subtypes. The project work was divided into 3 phases: (1) screening/identification by proteomic analysis and further characterization by RT-PCR and immunohistochemistry of the candidate protein; (2) clinical validation by quantitative ELISA assay of 57 (33 M, 24 F) PA patients and 50 normotensive controls (21 M, 29 F); (3) analysis of adrenal tissue of 8 individuals who had undergone adrenalectomy for APA or other adrenal tumors. The proteomic analysis showed a different expression of Serpin B3 Inhibitor-SCCA1 (SB3) in APA and BAH patients. Urine SB3 concentrations in normotensive controls, quantified by ELISA assay and normalized by urinary creatinine, resulted much lower in males (6.72 ng SB3 per mg creatinine, C.I. 4.43-10.19) than in females (20.56 ng SB3 per mg creatinine, C.I. 12.43-33.99, p < 0.00001). SB3 concentrations were not significantly different in males affected by different PA subtypes (BAH, n = 19 and APA, n = 14) compared with normotensive subjects (n = 21). In contrast, in PA females, SB3 was significantly higher in APA (n = 13) than in BAH patients (n = 11) or in normotensive controls (n = 29) (P < 0.01 and <0.05, respectively). Neither messenger RNA nor SB3 protein were identified in tissue obtained from adrenal tumors and from the surrounding normal gland. In conclusion urine SB3 concentrations are physiologically much lower in males than in females. Hypertensive women, affected by APA, present urinary SB3 concentrations significantly higher than women affected by BAH.

Aberrant cortisol regulations in bilateral macronodular adrenal hyperplasia: a frequent finding in a prospective study of 32 patients with overt or subclinical Cushing's syndrome.

CONTEXT: ACTH-independent macronodular adrenal hyperplasia (AIMAH) is a rare and heterogeneous condition characterized by abnormal steroid production. Cortisol secretion can be regulated by aberrant hormone receptors. OBJECTIVE: A large series of patients with AIMAH were evaluated to provide information on the prevalence and profile of aberrant regulations, in relation with the functional status. DESIGN AND PATIENTS: Thirty-two consecutive patients with AIMAH were prospectively studied: 10 had a Cushing's syndrome (CS), and 22 had a subclinical CS (SCS). METHODS: A baseline endocrine evaluation was followed by an in vivo protocol in search of aberrant cortisol responses (seven provocative tests). An acute inhibition test with the somatostatin analog octreotide was also performed. RESULTS: At least one aberrant cortisol response was identified in 28 of 32 (87%) patients. The overall prevalence of aberrant responses was independent of the functional status. Responses to the upright posture and to metoclopramide were frequently observed (67 and 56% respectively). A glucagon response was frequently observed in the SCS group (58%). A cortisol inhibition by octreotide was specifically found in the three CS patients who positively responded to the mixed meal, and was observed also in 12 of 13 (92%) patients with SCS. CONCLUSIONS: Cortisol responses indicative of aberrant receptor expression were highly prevalent in AIMAH. Thorough phenotyping of AIMAH may help uncover the underlying pathophysiology.

Adrenocorticotrophin-independent macronodular adrenal hyperplasia in a patient with lysine vasopressin responsiveness but insensitivity to gastric inhibitory polypeptide.

The aetiology of ACTH-independent macronodular adrenal hyperplasia (AIMAH) is uncertain. We examined a 55 year old man with Cushing's syndrome due to AIMAH, whose cortisol levels increased after stimulation with lysine-8-vasopressin (LVP) in vitro as well as in vivo. Abdominal MRI revealed nodular enlargement of both adrenal glands. No adenoma was evident on pituitary MRI. 131I-adosterol scintigraphy exhibited marked uptake into both adrenal glands. Although baseline plasma cortisol levels were within normal limits, urinary free cortisol excretion was 3-fold higher than the upper limit of the normal range. Plasma ACTH levels were undetectable. Oral dexamethasone failed to suppress plasma cortisol levels irrespective of dose, and administration of corticotrophin releasing hormone failed to increase plasma ACTH and cortisol levels. LVP injection failed to increase plasma ACTH levels, but elicited an increase in plasma cortisol levels. The direct stimulatory effect of LVP on cortisol secretion was confirmed in vitro in cultured adrenocortical cells from macronodules obtained at surgery. Food intake, gastric inhibitory polypeptide (GIP), or octreotide administration, which were reported to regulate cortisol release in patients with AIMAH, failed to affect plasma cortisol levels. In conclusion, plasma cortisol responsiveness to LVP, GIP, and octreotide is heterogeneous in patients with AIMAH.

Positive correlation between urinary excretion of catecholamine metabolites and tumour mass in pheochromocytoma. Results in patients with sustained and paroxysmal hypertension and multiple endocrine neoplasia.

In a series of 53 pheochromocytoma patients operated on at Sahlgren's Hospital during 1956-82, a positive linear correlation is established between the 24-hour urinary excretion of vanilmandelic acid (mumol/24 h) and tumour mass (g). A similar correlation was found between the excretion of metanephrines and tumour mass in 33 subjects. The patients were subgrouped according to their type of hypertension. A statistically significant correlation between vanilmandelic acid excretion and tumour mass persisted in groups IA (sustained hypertension without attacks), IB (sustained hypertension with attacks), and II (paroxysmal hypertension) but not in group III (miscellaneous patients). There was also a correlation between metanephrine excretion and tumour mass in groups IB (n = 8) and II (n = 12). In 10 patients with the syndrome of multiple endocrine neoplasia, a positive correlation was found between tumour mass and the excretion of vanilmandelic acid, metanephrines and adrenaline.

Development and preliminary application of a new assay for aldosterone stimulating factor.

A new assay method has been developed to measure aldosterone stimulating factor (ASF) quantitatively. This method utilizes a combination of affinity chromatography and high pressure liquid chromatography (HPLC). Antibodies raised against ASF, coupled to Affi-Gel 10, were used as the affinity column, and adsorbed ASF was eluted with 4 M urea. Quantitation was based on the external standard method of analysis using the area of the standards as controls, which were previously tested by HPLC for purity. The amount of ASF in the unknown samples was calculated on the basis of the area of the peak emerging at the retention time of ASF. A parallel bioassay using adrenal glomerulosa cells was also done to assess the biological activity. In eight normal volunteers, the urinary ASF was 146 +/- 4.6 ng/24 hr urine, whereas in plasma it was 70.5 +/- 6.5 ng/100 ml. The method is reproducible, specific for ASF, and showed less than 3% inter- or intraassay variability. In a preliminary study, when ASF was quantified from the 24-hour urine of two patients with adrenal hyperplasia, a significantly higher level of ASF was found (750 and 1020 ng/24 hr urine). These data suggest that ASF may be of pathological significance in certain hypertensive patients, especially in the hypertension associated with hyperaldosteronism.

Androgen excretion in women with a family history of breast cancer or with epithelial hyperplasia or cancer of the breast.

Urinary testosterone and androstanediol were measured by gas chromatography in four groups of premenopausal subjects: 22 healthy women (control group), 21 healthy women with a family history of breast cancer (familiality group), 39 patients with breast lumps which consisted of ductal or lobular hyperplasia (hyperplasia group) and 18 patients with infiltrating breast carcinoma (carcinoma group). On the basis of normal values found in our laboratory, steroid levels were above normal in 4.5% of the controls, 4.7% of the familiality group, 38.5% of the hyperplasia group (P less than 0.01 vs controls) and 61.1% of the carcinoma group (P less than 0.001 vs control group). The mean testosterone level in the carcinoma group (11.3 +/- 6.78 S.D.) and the mean androstanediol level in the hyperplasia group (47.25 +/- 31.0 S.D.) were significantly higher than those of the control group (testosterone 6.25 +/- 3.48 S.D., androstanediol 32.55 +/- 20.0 S.D.). No significant difference was found in mean testosterone or androstanediol levels between the control group and the familiality group (testerone 5.41 +/- 3.6 S.D., androstanediol 29.38 +/- 15.89 S.D.). We conclude that increased excretion of androgenic steroids is a hormonal abnormality common to breast cancer patients and to patients with breast epithelial hyperplasia, but not to subjects with a family history of carcinoma of the breast.

Deoxycorticosterone and aldosterone excretion in Cushing's syndrome.

We have used specific radioimmunoassays to measure urinary free deoxycorticosterone (DOC) and total aldosterone excretion in 15 patients with Cushing's syndrome. Free DOC excretion was increased in 6 of 8 patients with pituitary-dependent adrenal hyperplasia, in 2 of 3 patients with adrenal adenoma, and in all 4 cases with adrenal carcinoma. The most marked increase was noted in the adrenal carcinoma cases. Aldosterone excretion was high, normal, or low in each of the three types of Cushing's syndrome. The free DOC response to metyrapone in Cushing's syndrome due to adrenal adenoma was markedly different from that in patients with adrenal hyperplasia (pituitary-dependent) and may serve as a test to ascertain the etiology of the disorder. Correlations of free DOC and aldosterone excretion with free cortisol excretion, and their responses to the administration of metyrapone and dexamethasone were compatible with ACTH dependency in adrenal hyperplasia, autonomous production of steroids in adrenal adenomas and a chaotic steroidogenesis in adrenal carcinoma.

Effect of structure on the excretion of steroidal acids in human urine and their potential colorimetric assay as 17-oxogenic steroids.

Following the oral administration of tritiated coricosterone, cortisol, deoxycorticosterone and progesterone to human subjects, 14.9%, 12.8%, 3.4% and 2.1% of the dose was recovered in the acidic metabolite fraction of 48 h urines after conventional hydrolysis and solvent partition. Neutral and acidic 17-oxogenic steroids (17-OGS) excreted in 24 h urines were also measured with the Zimmermann color reaction. The acidic 17-IGS accounted for 14.1%, 16.4% and 12.1% of the fractionated 17-OGS in the urines of normal and pregnant females and normal males respectively. From structural considerations, it was concluded that only 17-hydroxylated steroidal acids, with a 20-hydroxy-21-oic acid side chain and derived predominantly from cortisol, would be estimated as 17-OGS after oxidation with sodium periodate. 17-Dexy steroidal acids resemble the neutral 17-deoxycorticosteroids in undergoing side chain oxication to 17-aldehydes which do not form Zimmermann chromogens. The excretion of both neutral and acidic 17-OGS was suppressible with dexamethasone administration.

Urine aldosterone radioimmunoassay: validation of a method without chromatography.

A simplified radioimmunoassay of urinary aldosterone is reported. Acid-hydrolyzed urine was extracted with dichloromethane and the extract assayed without further purification, Urinary aldosterone values in patients with Cushing's syndrome, low and normal-renin essential hypertension, congenital adrenal hyperplasia, and primary aldosteronism determined by this method agreed closely (r = 0.95, P less than 0.01) with values obtained using a standardized chromatographic method. This simplified assay represents a significant advance in out capabilitites for evaluating patients for abnormalities in aldosterone physiology.

Pregnanetriolone, a normal steroid metabolite: its excretion by normal, Cushing's syndrome and congenital adrenal hyperplasia subjects.

Synthesis of 3H-pregnanetriolone permitted the estimation of pregnanetriolone in urine with a sensitivity in excess of most previous claims. A good correlation (r = +0.97) was obtained between the values from gas liquid chromatography and those of a double isotope derivative method. In contrast to previous reports, these methods indicated that pregnanetriolone is excreted by normal adults. Urinary pregnanetriolone levels were 18-59 mug/24hr for normal subjects, 35-290mug/24hr in Cushing's syndrome and 250-7000 mug/24hr with congenital adrenal hyperplasia. It is concluded that pregnanetriolone is a normal steroid metabolite and its occurrence in Cushing's syndrome does not necessary indicate an abnormal steroid biosynthetic pathway.