Urinary Excretion Of Vanillylmandelic Acid Research Articles

ISOLATION OF TECOMINE FROM ALKALOID FRACTION, INVESTIGATION OF ADAPTOGENIC AND NEUROPSYCHOPHARMACOLOGICAL EFFECTSOF ETHANOLIC, ETHYL ACETATE AND HEXANE FLOWER EXTRACTS OF TECOMA GAUDICHAUDI

The present study aims to evaluate antistress and neuropharmacological parameters of ethanolic, ethyl acetate and hexane flower extracts of Tecoma gaudichaudi. Forced swim stress was used to evaluate antistress activity. The 24 h urinary excretion of vanillylmandelic acid (VMA) and ascorbic acid were determined by spectrophotometric methods in all groups under normal and stressed conditions. In the present study, the extracts (200 and 400 mg/kg, p.o.) were investigated for nootropic activity in rats with and without stress. Cook’s pole climbing apparatus measures conditioned avoidance response, elevated plus maze determines transfer latency and staircase test calculates the number of steps climbed and rearings in normal and stress induced rats to assess cognitive-improving activities. Cognition was enhanced by daily administration of Tecoma gaudichaudi extracts at doses of 200 and 400 mg/kg, p.o. in dose dependent manner in normal rats. Tecomine and its derivatives were isolated from alkaloidal fraction.

An Experimental Evaluation of Adaptogenic Potential of Standardized Epipremnum Aureum Leaf Extract.

Background:Stress is a normal part of everyday life but chronic stress can lead to a variety of stress-related illnesses including hypertension, anxiety, and depression. In the present investigation, standardized leaf extract of Epipremnumaureum was evaluated for its anti-stress potential.Materials and Methods:For the evaluation of anti-stress activity, groups of mice (n = 6) were subjected to forced swim stress and anoxic stress tolerance test in mice 1h after daily treatment of E.aureumextract. Diazepam (5 mg/kg) was taken as a reference standard. Urinary vanillylmandelic acid (VMA) and ascorbic acid were selected as noninvasive biomarkers to assess the anti-stress activity and plasma cortisol, blood ascorbic acid, and weight of adrenal were measured. The 24 h urinary excretion of VMA and ascorbic acid were determined by spectrophotometric methods in all groups under normal and stressed conditions. The hematological parameters (neutrophils, lymphocytes, and eosinophils) were also determined.Results:Administration of E.aureumat doses of 400 and 600 mg/kg wasfound to be effective in inhibiting the stress induced urinary biochemical changes in a dose-dependent manner. Treatment with E. aureum extract prevents the rise in blood ascorbic acid and plasma cortisol. Moreover, the extract prevented the increase in weight of adrenal gland also significantly increased the anoxia stress tolerance time. Dose-dependent significant reduction in white blood cell count was observed in anoxic stress tolerance test as compared to stressed group.Conclusion:Hence, the present study provides scientific support for the positiveadaptogenic effect of E. aureum extract.

Enhanced FGF23 production in mice expressing PI3K‐insensitive GSK3 is normalized by β‐blocker treatment

Glycogen synthase kinase (GSK)-3 is a ubiquitously expressed kinase inhibited by insulin-dependent Akt/PKB/SGK. Mice expressing Akt/PKB/SGK-resistant GSK3α/GSK3β (gsk3(KI)) exhibit enhanced sympathetic nervous activity and phosphaturia with decreased bone density. Hormones participating in phosphate homeostasis include fibroblast growth factor (FGF)-23, a bone-derived hormone that inhibits 1,25-dihydroxyvitamin D3 (1,25(OH)2D3; calcitriol) formation and phosphate reabsorption in the kidney and counteracts vascular calcification and aging. FGF23 secretion is stimulated by the sympathetic nervous system. We studied the role of GSK3-controlled sympathetic activity in FGF23 production and phosphate metabolism. Serum FGF23, 1,25(OH)2D3, and urinary vanillylmandelic acid (VMA) were measured by ELISA, and serum and urinary phosphate and calcium were measured by photometry in gsk3(KI) and gsk3(WT) mice, before and after 1 wk of oral treatment with the β-blocker propranolol. Urinary VMA excretion, serum FGF23, and renal phosphate and calcium excretion were significantly higher, and serum 1,25(OH)2D3 and phosphate concentrations were lower in gsk3(KI) mice than in gsk3(WT) mice. Propranolol treatment decreased serum FGF23 and loss of renal calcium and phosphate and increased serum phosphate concentration in gsk3(KI) mice. We conclude that Akt/PKB/SGK-sensitive GSK3 inhibition participates in the regulation of FGF23 release, 1,25(OH)2D3 formation, and thus mineral metabolism, by controlling the activity of the sympathetic nervous system.

Evaluation of enalapril affecting the renin-angiotensin system in normal and stress-induced rats based on urinary metabolites of amines and cortisol

The renin-angiotensin system is distributed centrally as well as peripherally, which works in association with SNS and HPA in stress. Angiotensin is considered one of the principal hormones for bringing out stress-related changes. To find the influence of enalapril on the neuromediator of CNS and periphery, the present work was carried out. The assessment of the transmitter levels was done by estimating major urinary stress metabolites like vanillylmandelic acid, 5-hydroxyindoleacetic acid, 6-β-hydroxycortisol and homovanillic acid in normal and stress-induced conditions and also in the absence and presence of enalapril treatment in rats. Groups of rats were subjected to a forced swim stress one hour after daily treatment of enalapril. The 24 h urinary excretion of vanillylmandelic acid, 5-hydroxyindoleacetic acid, 6-β-hydroxycortisol and homovanillic acid was determined in all groups under normal and stressed-induced conditions. Daily administration of enalapril at doses of 3, 10 and 30 mg/kg/body weight inhibited the stress-induced urinary biochemical changes. However, treatment with enalapril in normal conditions increased the excretion of all selected metabolites. It can be speculated that enalapril prevented stress-induced increases of adrenergic, serotonergic and cortical mechanisms.

Enhanced catecholamine release in mice expressing PKB/SGK-resistant GSK3

Glycogen synthase kinase 3 (GSK3) plays a decisive role in the regulation of multiple functions. GSK3 is phosphorylated and its activity inhibited by protein kinase B (PKB/Akt) and serum and glucocorticoid inducible kinase (SGK) isoforms, which are in turn activated by growth factors through phosphoinositide (PI) 3 kinase signaling. PI3/PKB/Akt/SGK-dependent inhibition of GSK3 is disrupted in gene-targeted knockin mice with mutated and thus PKB/SGK-resistant GSK3α,ß (gsk3 ( KI )) where the serine of the PKB/SGK phosphorylation site has been replaced by alanine. Recent experiments revealed that blood pressure is significantly higher in those mice than in wild type mice (gsk3 ( WT )). The present study was performed to elucidate the underlying cause. Blood pressure was determined with the tail cuff method, heart rate by ECG measurements, catecholamine concentrations by ELISA, and vanillylmandelic acid by high pressure liquid chromatography. As a result, blood pressure and heart rate were significantly higher in gsk3 ( KI ) than in gsk3 ( WT ) mice. The α-adrenergic blocker prazosin (1μg/g body weight, b.w.) and the ganglion blocker hexamethonium (40μg/g b.w.) decreased blood pressure to a larger extent in gsk3 ( KI ) than in gsk3 ( WT ) mice and virtually abrogated the difference between genotypes. Similarly, the β-adrenergic blocker atenolol (5μg/g b.w.) decreased the heart rate to a larger extent in gsk3 ( KI ) than in gsk3 ( WT ) mice and again dissipated the difference of heart rate between genotypes. Plasma epinephrine and norepinephrine concentrations, as well as urinary excretion of vanillylmandelic acid, were significantly higher in gsk3 ( KI ) than in gsk3 ( WT ) mice. The observations reveal a completely novel function of PKB/Akt/SGK-dependent GSK3 signaling, i.e., regulation of catecholamine release.

Adaptogenic and nootropic activities of aqueous extract of Carum carvi Linn. (Caraway) fruit: An experimental study in Wistar rats

In the present investigation the aqueous extract of Carum carvi was evaluated for antistress activity in normal and stress induced rats. The extract was studied for nootropic activity in rats and in vitro antioxidant potential to correlate its antistress activity. For the evaluation of antistress activity groups of rats were subjected to forced swim stress one hour after daily treatment of Carum carvi extract. Urinary vanillylmandelic acid (VMA) and ascorbic acid were selected as non invasive biomarkers to assess the antistress activity. The 24 h urinary excretion of vanillylmandelic acid (VMA) and ascorbic acid was determined in all groups under normal and stressed conditions. The nootropic activity of the extract as determined from acquisition, retention and retrieval in rats was studied by conditioned avoidance response using Cook’s pole climbing apparatus. The in vitro antioxidant activity was determined based on the ability of Carum carvi to inhibit lipid peroxidation in liver and brain homogenates. Daily administration of Carum carvi at doses of 100, 200 and 300 mg/kg body weight one hour prior to induction of stress inhibited the stress induced urinary biochemical changes in a dose dependent manner. However no change in the urinary excretion of VMA and ascorbic acid was observed in normal animals at all the doses studied. The cognition, as determined by the acquisition, retention and recovery in rats was observed to be dose dependent. The extract produced significant inhibition of lipid peroxide formation in comparison with ascorbic acid in a dose dependent manner in both liver and brain. The present study provides scientific support for the antistress (adaptogenic), antioxidant and nootropic activities of Carum carvi extract and substantiates its traditional use as a culinary spice in foods as beneficial and scientific in combating stress induced disorders.

HPLC–mass spectrometry method for quantitative detection of neuroendocrine tumor markers: Vanillylmandelic acid, homovanillic acid and 5-hydroxyindoleacetic acid

Background The urinary excretion of vanillylmandelic acid (VMA), homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) can be increased in the presence of neuroblastic and carcinoid tumors. The former is characterized by defective catecholamine metabolism which results in high urinary levels of VMA and HVA. The latter shows an altered metabolism of tryptophan and an increased synthesis of serotonin, producing high 5-HIAA urinary concentrations. Methods We describe an HPLC-tandem mass spectrometric method for the simultaneous quantification of VMA, HVA and 5-HIAA in human urine. The chromatographic separation is performed on a reversed-phase C18 column. Instrumental analysis is performed on a Q-Trap 2000 triple quadrupole/ion trap mass spectrometer. Results The method is fast and does not require sample pre-treatment. Multiple calibration curve exhibited consistent linearity and reproducibility. Linear responses were observed in the concentration range 0–50 mg/l for each analyte. Limits of detection were 0.001 mg/l for VMA, 0.015 mg/l for 5-HIAA and 0.050 mg/l for HVA with a signal-to-noise ratio of 3. Limits of quantification were 0.005 mg/l for VMA, 0.050 mg/l for 5-HIAA and 0.1 mg/l for HVA with a signal-to-noise ratio of 10. Conclusions This method can be proposed as a tool for neuroendocrine tumor markers detection.

Adaptogenic and nootropic activities of aqueous extract of Vitis vinifera (grape seed): an experimental study in rat model

BackgroundThe aerial parts of Vitis vinifera (common grape or European grape) have been widely used in Ayurveda to treat a variety of common and stress related disorders. In the present investigation, the seed extract of V. vinifera was evaluated for antistress activity in normal and stress induced rats. Furthermore, the extract was studied for nootropic activity in rats and in-vitro antioxidant potential to correlate its antistress activity.MethodsFor the evaluation of antistress activity, groups of rats (n = 6) were subjected to forced swim stress one hour after daily treatment of V. vinifera extract. Urinary vanillylmandelic acid (VMA) and ascorbic acid were selected as non-invasive biomarkers to assess the antistress activity. The 24 h urinary excretion of vanillylmandelic acid (VMA) and ascorbic acid were determined by spectrophotometric methods in all groups under normal and stressed conditions. The nootropic activity of the extract as determined from acquisition, retention and retrieval in rats was studied by conditioned avoidance response using Cook's pole climbing apparatus. The in vitro antioxidant activity was determined based on the ability of V. vinifera to scavenge hydroxyl radicals.ResultsDaily administration of V. vinifera at doses of 100, 200 and 300 mg/kg body weight one hour prior to induction of stress inhibited the stress induced urinary biochemical changes in a dose dependent manner. However, no change in the urinary excretion of VMA and ascorbic acid was observed in normal animals at all the doses studied. The cognition, as determined by the acquisition, retention and recovery in rats was observed to be dose dependent. The extract also produced significant inhibition of hydroxyl radicals in comparison to ascorbic acid in a dose dependent manner.ConclusionThe present study provides scientific support for the antistress (adaptogenic), antioxidant and nootropic activities of V. vinifera seed extract and substantiate the traditional claims for the usage of grape fruits and seeds in stress induced disorders.

Use of Piritramide for Analgesia and Sedation during Peribulbar Nerve Block for Cataract Surgery

Purpose: To investigate the effects of pre-block analgesia and sedation using piritramide on haemodynamic stability, endocrine stress response and patients’ pain perception. Methods: In a randomized, single-blinded, placebo-controlled study, 60 patients having cataract surgery with peribulbar block were randomly assigned into two groups: group A (n = 30) received 0.05 mg/kg piritramide (Dipidolor^®) intravenously; group B received normal saline intravenously prior to peribulbar block. Mean arterial pressure, heart rate, respiratory rate and pulse oximetry were recorded perioperatively. Pain from peribulbar block was assessed using a verbal analogue scale. Urinary excretion of vanillylmandelic acid was measured to assess the endocrine stress response. Using a questionnaire, patients assessed their anxiety and back pain before and during surgery. Results: Mean arterial pressure remained near baseline in group A. In group B, a significant increase in mean arterial pressure after peribulbar block was found (p < 0.001). In addition, a significant increase in urinary excretion of vanillylmandelic acid was found in group B (p = 0.013). Pain scores (p < 0.001), anxiety before nerve block (p = 0.02) and during surgery (p < 0.001) and back pain (p = 0.003) were significantly lower in group A. Conclusion: The presented study suggests that using piritramide for analgesia and sedation prior to peribulbar block produces haemodynamic stability and reduces pain perception and endocrine stress response.

Recurrent melanotic neuroectodermal tumor of infancy

(Editorial Comment: I view that adjuvant therapy may be necessary to control residual or recurrent disease in this unusual tumor.) Melanotic neuroectodermal tumor of infancy (MNTI) is an uncommon neoplasm found primarily in children less than one year of age. First described in the early 1900s by Krompecher as a congenital melanocarcinoma, 1 the nomenclature of this neoplasm has undergone multiple changes that have been based on histologic and biochemical studies. Since Borello and Gorlin’s discovery of the associated high urinary excretion of vanillylmandelic acid (VMA) in the late 1960s, 2 MNTI has been thought to be a derivative of neural crest cells. More recently, Hoshino et al have demonstrated high levels of urine catecholamines in a case of MNTI, 3 which further supports the neural origin of this rare neoplasm. In a review of the world literature, approximately 227 cases of MNTI have been reported, most of which (93%) involve the head and neck region. The maxilla (69%) is the most commonly affected site, followed by the skull (11%), the mandible (6%), and less commonly, brain, mediastinum, thigh, epididymis, foot, and shoulder. 4 This tumor is reported to be malignant in only 4% of the cases 5 but is considered to be locally aggressive.

CYSTIC PARAGANGLIOMA OF THE RENAL CAPSULE

A 42-year-old black woman from Ghana was referred for evaluation of recurrent pain in the left flank. Physical examination revealed a voluminous palpable mass in the left upper abdomen. Laboratory values, including catecholamine levels (urinary vanillylmandelic acid excretion per 24 hours), were normal except for peripheral eosinophilia. The patient had no hypertension. Computerized tomography showed an 11 cm. left cystic renal mass of the lower pole (fig. 1). Intravenous contrast injection revealed that the mass had thickened irregular septa running through it. Imaging studies did not afford an unequivocal interpretation. However, peripheral eosinophilia and the fact that the patient was of African origin led us to suspect a renal hydatid cyst. Intraoperatively the tumor was well demarcated from the renal parenchyma and surrounding organs and enclosed by the renal capsule. Therefore, it was extirpated, leaving the renal parenchyma intact. Grossly the tumor was well circumscribed and multilocular, and contained abundant hemorrhagic fluid. Histologically, the tumor was composed of small monomorphic spindle cells and was strongly immunoreactive for chromogranin, vimentin and neuron specific enolase but not for cytokeratins (fig. 2). Pathological features were consistent with a cystic extra-adrenal paraganglioma. Scintigraphic imaging with 131 iodine-metaiodobenzylguanidine for evaluation of metastatic disease was negative. No additional treatment was given. The patient was well at 10-month followup. DISCUSSION

Pheochromocytoma: Clinical Observations from a Brooklyn Tertiary Hospital

ObjectiveTo report eight cases of pheochromocytoma, diagnosed and treated at our tertiary hospital during a 42-month period. MethodsWe review clinical manifestations as well as historical and family data. Biochemical and radiologic methods for diagnosis and tumor localization at our institution were compared with methods at other teaching centers. ResultsAmong 4,180 patients with essential hypertension encountered during a 42-month interval, 8 had pheochromocytoma. The most common clinical findings were diaphoresis, chest or abdominal pain, palpitations, headaches, and nausea. Clinical features were remarkable only for the significant correlation between urinary excretion of vanillylmandelic acid and tumor volume (r = 0.925; P<0.01). Only one patient, with a history of medullary thyroid carcinoma, had multiple endocrine neoplasia. No unusual familial concentration of pheochromocytomas was found in our community, and our diagnostic techniques were similar to those reported from other institutions. We calculated an average annual incidence rate of 0.5 per 100,000 person-years, a figure comparable to that reported at the Mayo Clinic in Rochester, Minnesota. ConclusionBecause our institution does not see the volume of referral patients as does the Mayo Clinic, we suggest that the frequency of pheochromocytoma may be underestimated in the patient population that we serve. (Endocr Pract. 2000;6:249-252)

Natural course of adrenal incidentalomas: seven-year follow-up study.

Most adrenal incidentalomas are nonfunctioning cortical adenomas that do not require surgery. Operative treatment is indicated if the incidentaloma has malignant features as seen during the radiologic workup, is hormonally active, or grows during follow-up. A conservative attitude is justified in asymptomatic patients if we know that the natural course of these tumors is not disadvantageous. We followed a group of patients treated by observation to determine the natural course of adrenal incidentalomas. Altogether 30 incidentalomas in 27 patients were detected in the Helsinki University Central Hospital from June 1981 through December 1992 and were re-examined during 1997. The patients were evaluated clinically, and hormonal testing was done by performing a 1-mg overnight dexamethasone suppression test; the next day we measured the 24-hour urinary excretion of vanillylmandelic acid, metanephrines and normetanephrines, and serum potassium. Magnetic resonance imaging (MRI) was used to determine the size of the tumor. If the patient had died during the follow-up period, the causes of death from the death certificates and autopsy reports were reviewed. The mean follow-up was 7 years. Nine patients had died, with none of deaths related to the incidentaloma. Sixteen patients were evaluated clinically and by the basic hormonal tests and MRI; none showed any signs of hormonal activity or features of malignancy. Two patients did not want to be reexamined but stated that they were asymptomatic. This follow-up study supports the conclusion that conservative therapy is justified because the growth tendency of adrenal incidentalomas is slow and they seem to remain hormonally inactive.

Acidic Catecholamine Metabolites and 5-Hydroxyindoleacetic Acid in Urine: The Influence of Diet

Concentrations of vanillylmandelic acid (VMA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), vanillic acid (VA) and 5-hydroxyindoleacetic acid (5-HIAA) in urine from healthy subjects were determined by a high-performance liquid chromatography system with a mixed-mode (C18/anion-exchange) column and an 8-channel electrochemical detector, in order to study the influence of diet, diurnal variation and age. The urinary excretion of 5-HIAA increased significantly after eating banana, pineapple, tomato, kiwi fruit and walnut. An increase in the urinary excretion of DOPAC and HVA after eating banana and that of VA after taking vanilla was also noted. The urinary excretion of VMA was not significantly influenced by any of the foods examined. The urinary excretion of 5-HIAA in the first-morning urine increased beyond the upper limit of the reference value when banana was taken at 2000 h the previous day. The excretion of all metabolites in the second-morning urine in the fasting state was within respective reference ranges. Diurnal variation of the excretion of VMA, DOPAC, HVA and 5-HIAA in urine was relatively small, but that of VA was large. The concentrations (mmol/mol creatinine) of VMA, DOPAC, HVA, 5-HIAA and VA in the first-morning urine from healthy subjects increased from 7 days after birth to 1 year and then decreased to adult levels at 13 years of age.

N-myc oncogene and urinary catecholamines in children with neuroblastoma.

This paper reports the analysis of N-myc amplification, urinary vanillylmandelic acid (VMA), and norepinephrine (NE) excretion and survival in 22 children with neuroblastoma and 7 with ganglio-neuroblastoma. Five patients had N-myc amplification (from 30 to more than 200 copies), all of whom had advanced-stage disease. The urinary excretion of VMA was normal in all of them, only one showed increased NE excretion. All patients with stage C-D disease and one copy of N-myc had increased VMA urinary excretion and increased (9/14) or normal (5/14) NE urinary excretion. All patients with stage A or B disease had one copy of N-myc. Half of them showed increased VMA urinary levels, while only 3/10 showed increased NE urinary values. Comparison of cumulative survival curves in relation to N-myc amplification and to VMA and NE urinary excretion showed a clear parallelism. Amplification of N-myc corresponded to normal VMA and NE urinary excretion, and was associated with the worst prognosis (P < 0.01), while increased VMA and/or NE excretion was found in patients with only one copy of N-myc.

Effect of glutamine-supplemented intravenous nutrition on survival after Escherichia coli-induced peritonitis.

Current solution formulations for total parenteral nutrition (TPN) do not contain glutamine (GLN). The purpose of this study was to examine whether GLN supplementation of TPN would improve survival in experimental Escherichia coli peritonitis in Fischer 344 rats (190-210 g). Initial experiments were performed to determine the degree of stress and to evaluate survival after intraperitoneal E coli injection. The E coli colony used was isolated from a culture of human blood. Graded doses were injected intraperitoneally in Fischer 344 rats (190-210 g). The response of white blood cell count, plasma insulin, glucagon, and corticosterone levels, and urinary excretion of vanillylmandelic acid reflected a significant stress response for at least 3 days. Survival was dose-dependent, with 60% mortality at 3 days after injection of 5 x 10(5) colony forming units of E coli/200 g body weight. To determine whether GLN supplementation of TPN would alter survival in this E coli peritonitis model, Fischer 344 rats were randomized to receive TPN containing 4.25% standard amino acids (group STD, n = 38) or the same solution with 1.5% of the amino acid content replaced with L-GLN (group GLN, n = 38). After 7 days of TPN, 5 x 10(5) colony forming units of E coli/200 g body weight were injected intraperitoneally under direct vision through a small laparotomy. Survival was monitored for 3 days. Surviving rats were killed to determine various nutritional parameters including plasma albumin and GLN concentration, the weight and nitrogen content of the gastrocnemius muscle, and biochemical and histological composition of the small intestine.(ABSTRACT TRUNCATED AT 250 WORDS)

Disseminated neuroblastoma (stage IV and IV-S) in the first year of life. Outcome related to age and stage. Italian Cooperative Group on Neuroblastoma.

Infants (age 0-11 months) with disseminated neuroblastoma are known to have a better prognosis than older children with the disease, but there is little information regarding factors that influence the outcome of the disease in these patients. The authors report a series of 110 infants with disseminated neuroblastoma with disease diagnosed between March 1976 and February 1991 in 21 institutions participating in the Italian Cooperative Group on Neuroblastoma (ICGNB). Of the 110 infants, 34 had Stage IV disease, and 76 had Stage IV-S disease. The 5-year survival probability was 77% for all patients, 71% for those with Stage IV disease, and 81% for those with Stage IV-S disease. Of the 34 infants with Stage IV disease, the 9 who were 5 months or younger at the time of disease diagnosis are all alive (1 with active disease) at 7-143 months after diagnosis, whereas of the 25 infants who were 6-11 months of age at the time of disease diagnosis, 10 have died. Of the 76 infants with Stage IV-S disease, 12/64 who were 5 months of age or younger at the time of disease diagnosis died (mostly of massive hepatomegaly); 9 of these deaths occurred in infants with disease diagnosed before they were 2 months old, whereas 1 death occurred in the 12 infants with disease diagnosed when they were 6-11 months old. Four infants with Stage IV-S disease achieved complete disease remission and subsequently had relapse of disease. High levels of serum LDH and low urinary excretion of vanillylmandelic acid were associated with worse prognosis. The authors suggest that infants older than 6 months of age who have Stage IV disease require aggressive therapy. For infants with disease diagnosed before they are 2 months old, Stage IV-S disease may have a worse prognosis than Stage IV disease.

Cytogenetic and Molecular Evaluation of Clinically Aggressive Esthesioneuroblastoma

We report a 16-year-old boy with esthesioneuroblastoma that presented with a unilateral tumor extending to the maxillary sinus and periorbital region. Despite initial therapy with gross resection, 5,682 cGy to the tumor bed and chemotherapy, the patient subsequently had a rapid local recurrence with distant metastases. Immunocytochemical, ultrastructural, cytogenetic, and molecular techniques were performed to determine if this tumor was biologically similar to childhood neuroblastoma. Urinary excretion of vanillylmandelic acid (VMA) and homovanillic acid (HVA) were markedly elevated. Chromogranin and neuron specific enolase immunostaining of tumor cells was positive, as seen in neuroblastoma. Electron microscopic studies showed cells that were closely packed and connected by occasional cell junctions. The cell cytoplasm contained moderate amounts of filaments and microtubules. Numerous electron dense granules were observed; however, these granules lacked distinct nucleoids and generally reacted strongly for acid phosphatase, indicating a lysosomal rather than a secretory function. Tumor cells contained near-pseudotetraploid chromosomes, with all chromosomes represented at least three times, and chromosome 5 was present in multiples of eight. Clonal structural abnormalities included 2q+ and 5q+ and multiple double minutes. Northern blot analysis revealed both c-myc and N-myc expression; however, N-myc amplification was not demonstrated, and c-myc expression appeared increased, unlike cases of rapidly progressive neuroblastoma. These results suggest that despite biologic similarities to neuroblastoma in catecholamine excretion and some ultrastructural features, molecular genetic abnormalities differ in this comparatively aggressive case of estesioneuroblastoma.

Chromogranin A Storage and Secretion

Chromogranin A, co-stored and co-released with catecholamines from adrenal medullary and sympathetic neuronal vesicles, is elevated in the plasma of patients with pheochromocytoma. The usefulness of the hormone in the differential diagnosis of hypertension is examined. An elevated level of chromogranin A had comparable diagnostic sensitivity (83%, 24/29) to, but greater diagnostic specificity (96%, 86/90) than the level of plasma catecholamines when subjects with pheochromocytoma (n = 29) were evaluated in comparison to several reference groups, including normotensive controls (n = 49), subjects with essential hypertension (n = 28), subjects with renovascular hypertension (n = 5), and subjects with primary aldosteronism (n = 3). Subjects with signs or symptoms suggesting pheochromocytoma, but in whom the diagnosis was ultimately ruled out (n = 5) had normal plasma levels of chromogranin A. A modest rise in chromogranin A in those with essential hypertension, and correlation of chromogranin A with diastolic blood pressure in normotensive patients and patients with essential hypertension did not impair the diagnostic usefulness of chromogranin A for pheochromocytoma. Renal failure was associated with an elevated plasma chromogranin A independently of blood pressure. Plasma chromogranin A correlated with tumor mass, tumor chromogranin A content, tumor norepinephrine content, and urinary vanillylmandelic acid excretion; it did not correlate with plasma or urinary catecholamines, nor with blood pressure in patients with pheochromocytoma. Plasma chromogranin A levels did not differ in subjects with pheochromocytoma when stratified by age, sex, tumor location, or tumor pathology. Several drugs used in the diagnosis or treatment of pheochromocytoma (clonidine, metoprolol, phentolamine, and tyramine) had little effect on plasma chromogranin A concentration. Within the pheochromocytoma, chromogranin A was localized along with catecholamines to the soluble core of chromaffin granules, where it accounted for 18 +/- 5% of vesicle soluble protein. We conclude that 1) chromogranin A emerges along with catecholamines from pheochromocytoma chromaffin granules; 2) plasma chromogranin A is a sensitive and specific diagnostic tool in evaluation of actual or suspected pheochromocytoma; 3) plasma chromogranin A predicts pheochromocytoma tumor size and overall catecholamine production; and 4) drugs commonly employed in the diagnosis or treatment of pheochromocytoma have little effect on plasma chromogranin A level, preserving the usefulness of chromogranin A in evaluating pheochromocytoma. Thus, measurement of chromogranin A provides a useful adjunct to the diagnosis of pheochromocytoma.

Phaeochromocytoma--investigation and management of 10 cases.

Since 1960 we have diagnosed phaeochromocytoma (paraganglioma) in 10 children. The cases include a 15 year old girl who over a three year period presented with multiple paragangliomata and an associated malignant carotid body tumour. All children were hypertensive, eight of 10 presenting with severe headaches. Diagnosis was based on finding a raised urinary vanillylmandelic acid excretion and plasma noradrenaline concentration. In addition six of eight children were hypercalcaemic with raised plasma calcitonin concentrations; plasma parathyroid hormone concentrations were high in two of seven and four out of eight children had raised plasma renin activities on presentation. No child, however, was found to have a multiple endocrine neoplasia syndrome. Despite the introduction of newer techniques for the detection of catecholamine producing tumours we found that selective arteriography and venous catecholamine sampling were superior for tumour localisation compared with ultrasound scanning, computed tomography, and metaiodo-benzyl-guanidine (MIBG) scanning.