Genomic and clinical characterization of a familial GIST kindred intolerant to imatinib.

Familial gastrointestinal stromal tumors (GIST) are rare. We present a kindred with multiple family members affected with multifocal GIST who underwent whole genome sequencing of the germline and tumor. Affected individuals with GIST harbored a germline variant found within exon 13 of the KIT gene (c.1965T>G; p.Asn655Lys, p.N655K) and a variant in the MSR1 gene (c.877 C > T; p.Arg293*, pR293X). Multifocal GISTs in the proband and her mother were treated with preoperative imatinib, which resulted in severe intolerance. The clinical features of multifocal GIST, cutaneous mastocytosis, allergies, and gut motility disorders seen in the affected individuals may represent manifestations of the multifunctional roles of KIT in interstitial cells of Cajal or mast cells and/or may be suggestive of additional molecular pathways which can contribute to tumorigenesis.

Plasma levels of atrial natriuretic factor in nonhibernating and hibernating marmots.

Plasma atrial natriuretic factor (ANF) was measured in 16 marmots at various times of the year. Nonhibernating males (n = 6) had an average plasma concentration of 56 +/- 8 pg/ml; nonhibernating females (n = 6) had an average plasma concentration of 61 +/- 4 pg/ml. During hibernation an additional group of females (n = 4) showed an average of 25 +/- 5 pg/ml. Plasma ANF of both groups of nonhibernating marmots was significantly higher (P less than 0.01) than that the hibernating group, but there was no difference between nonhibernating males and females.

Thickness of medial layers of arteries prior to and during hibernation.

The thickness of the media of the aorta and the carotid, iliac, and renal arteries was measured in hibernating and nonhibernating marmots. A positive correlation was found between body weight and the thickness of the media of carotid and iliac arteries. The relation noted apparently was not due to the mean arterial pressure prior to and during hibernation, since there was no difference in the thickness of the media between nonhibernating and hibernating females. The difference due to gender could be accounted for by the greater average body weight of males. The media of the renal arteries demonstrated no relation between their thickness and body weight.

Effects of anesthetics on cardiovascular responses of the marmot Marmota flaviventris.

The effects of pentobarbital (30 mg/kg), urethan (2 g/kg), chloralose/urethan (50 mg/kg, 500 mg/kg), and thiobutabarbital (Inactin, 100 mg/kg) on the mean arterial pressure (BP) and heart period (HP) of Marmota flaviventris were examined. Anesthesia significantly decreased BP by 22-27 mm Hg and HP by 123-151 msec. In a series of paired studies with eight marmots it was found that pentobarbital increased the BP response to phenylephrine and almost abolished the baroreflex HP responses to phenylephrine and nitroglycerin. In another series of animals right carotid occlusion in unanesthetized animals produced greater changes in BP and HP than occlusion of the left carotid. Chloralose/urethan, urethan, or Inactin reduced the reflex BP response to unilateral carotid occlusion by 50% and the HP response by 96%. It was concluded that the anesthetic agents investigated depress baroreflex responses significantly by influencing efferent sympathetic and parasympathetic reflex responses. They, therefore, are not appropriate for cardiovascular studies in acute, anesthetized preparations of the marmot and, perhaps, other hibernating species.

Seasonal variation of cardiovascular function in the marmot, Marmota flaviventris.

Monthly measurements of heart rate, mean arterial pressure, and cardiac output were made on active and hibernating marmots from the time of emergence from hibernation through the next hibernation period. From these measurements cardiac index, stroke index, and total peripheral resistance were calculated on the basis of estimated lean body mass. Heart rate was low after emergence (132 +/- 9.5 beats (B)/min), peaked in August (160 +/- 9.3 B/min), and then fell slightly in September and October. During hibernation heart rate fell to 9 +/- 1.1 B/min. Mean arterial pressure, which was low in early spring (101 +/- 6.9 mm Hg), rose to a peak value in June (131 +/- 7.7 mm Hg) and remained essentially unchanged until hibernation when it fell to 52 +/- 4.0 mm Hg. Cardiac index (61 +/- 4.9 ml/kg min) in March rose to a peak in May (83 +/- 8.5 ml/kg min) and fell linearly until October. There was an additional drop in cardiac index during hibernation (7.6 +/- 0.9 ml/kg min). Total peripheral resistance increased linearly from the time of emergence until October. Most of this change was due to the decrease in cardiac index. Stroke index showed no significant changes in the prehibernation period, but increased by 55% during hibernation. Maintenance of arterial pressure in the months preceding hibernation in the face of diminishing cardiac index indicate that alterations in vasomotor tone or shifts in patterns of blood flow occurred prior to the hibernation period.

Seasonal changes of food and water consumption and urine production of the marmot, Marmota flaviventris.

In early spring, food and water consumption and the excretion and clearances of urine and solutes reached maximal rates. Water consumption exceeded food intake and urine production and plasma osmolality was lowest. Toward early and late summer, water intake decreased faster than food consumption and urine production. Urea excretion and clearances diminished with food consumption, while creatinine clearance decreased only slightly. Plasma osmolality increased. The data are consistent with rehydration soon after hibernation is completed, followed by a period of weight gain and dehydration in preparation for the next prolonged period of hibernation.

Factors that alter the plasma renin activity of the marmot.

The effects of low doses of norepinephrine (NE) and furosemide and a volume load (nonhibernators only) on plasma renin activity (PRA), mean arterial pressure (MAP), heart rate (HR), left renal (RBF) and right iliac (IBF) blood flow, cardiac index (CI), and total peripheral resistance (TPR) were determined in euthermic and hibernating marmots. In nonhibernating marmots NE produced an increase in CI and TPR and a decrease in RBF. In hibernators this dose of NE caused an increase in MAP, HR, and renal resistance, whereas it decreased PRA and did not alter iliac resistance. Furosemide infusions led to an increase in PRA in both groups and an increase in TPR in nonhibernators. The volume load in nonhibernators produced only a decrease in PRA. A comparison of control data from the two groups indicated that the renal and iliac beds contribute only a small portion to the increase in TPR that occurs during hibernation.

Renal function of the pony and the horse.

Simultaneous renal clearances of inulin (CIN), p-aminohippurate (CPAH), and creatinine (CCR) were measured in hydrated mares (6 ponies and 2 horses). The CIN and CPAH were determined during steady-state infusion at 3 different infusion rates. A 6-fold change in plasma IN concentration did not produce alteration in CIN, nor was there a difference between the ponies and horses (P greater than 0.2). The overall average (mean +/- SEM) was 190.6 +/- 5.89 ml . min-1 . 100 kg of body weight-1. There was no difference noted between simultaneous CIN and CPAH. Clearance of PAH remained essentially constant during the change in plasma PAH from 0.33 mg/dl to 5.27 mg/dl. The extraction ratio of PAH for the nonanesthetized pony was 0.966. Effective renal plasma flow (CPAH) of the pony exceeded that of the horse.

Circannual renal function and plasma electrolytes of the marmot.

Studies with unanesthetized active (non-lethargic) marmots demonstrated circannual rhythms of renal function (clearances of p-aminohippurate, creatinine, and inulin), plasma osmolality, and plasma sodium and potassium concentrations. Effective renal plasma flow, glomerular filtration rate, the clearance of creatinine, and plasma potassium were highest in spring and lowest in the fall and winter. Plasma osmolality and plasma sodium concentrations tended to be highest during the winter months and lowest in the spring and summer. p-Aminohippurate excretion was shown to be a Tm-limited system.

Maintenance of ion concentration gradients in the cold in aorta from rat and ground squirrel.

Cell water and ionic content were measured in aortic smooth muscle from rats and ground squirrels during 48 h of incubation in oxygenated Krebs solution held at low temperatures. Cells from the ground squirrel, a hibernator, maintained sodium and potassium contents near normal levels during incubation at 7 degrees C. In sharp contrast, cells from the rat lost potassium and gained sodium with half times of 14 and 11 h, respectively. The [K] in cell water for the rat decreased 138 mM while [Na] in cell water increased about 148 mM. Cells from the hibernator tended to lose 10 mM K and gain 13 mM Na. Vascular smooth muscle of both the rat and squirrel was able to maintain K content at 17 degrees C. Tissue content of the divalent cations Ca and Mg did not change in vessels from either animal. The present findings suggest that aortic smooth muscle cells from a hibernator have adapted in such a way as to maintain ionic gradients upon prolonged exposure to low temperatures.

Effect of intravenous infusion of ethanol on plasma glucose, lactic acid and pyruvic acid concentrations in miniature swine.

Fructose, infused intravenously with ethanol, alleviated ethanol-induced hypoglycemia in adult miniature swine at all concentrations tested. Increases in plasma lactic acid and pyruvic acid concentrations also were found.

Renin-angiotensin-aldosterone system of the normothermic marmot.

Adrenal steroid secretion rates and the renin-angiotensin-aldosterone (RAA) system were studied in the normothermic marmot. Adrenal secretion by the anesthetized, laparotomized marmot was (mean +/- SEM); aldosterone 1.2 +/- 0.3 ng/min, deoxycorticosterone 16.7 +/- 11.5 ng/min, corticosterone 15.2 +/- 7.8 ng/min, and cortisol 554 +/- 108 ng/min. Four forcings were investigated that affect feedback control at different sites: adrenocorticotropic hormone (ACTH) and angiotensin II (AII) infusion, sodium (Na) depletion, and Na loading. Plasma aldosterone, cortisol, Na, and potassium (K) concentrations as well as plasma renin activity (PRA) hematocrit (Hct), and in some studies, blood pressure were measured. ACTH infusion increased the plasma concentrations of aldosterone and cortisol. AII infusion increased aldosterone concentration, blood pressure, and Hct. Na depletion increased aldosterone, Hct, and PRA; plasma Na and K were decreased. Aldosterone concentration, Hct, and PRA decreased after salt loading. Normothermic, salt-depleted marmots demonstrated a pronounced fall in blood pressure following infusion of the AII analog, 1-sarcosine-8-alanine AII. The average plasma values for aldosterone, PRA, and cortisol found in 44 control animals were: aldosterone 3.8 +/- 0.3 ng/100 ml, PRA 1.9 +/- 0.2 ng AI-ml-1-h-1, and cortisol 54 +/- 4 ng/ml. It was concluded that normothermic marmots have a RAA system comparable to other mammalian species.

The renin-angiotensin-aldosterone system in rabbits with congestive heart failure produced by aortic constriction.

Severe constriction of the suprarenal abdominal aorta of 3-kg rabbits to 3.7+/-0.2 mm2 and maintenance of a daily sodium intake of 10 mE q by infusion of 0.9% sodium chloride resulted in a progressive increase in central ear arterial pressure to 106+/-3 (SEM) mm Hg (control=79+/-1). This was accompanied by a progressive increase in left ventricular end-diastolic pressure to 22+/-2 mm Hg (control=3+/-1), plasma renin activity to 21+/-5 ng of angiotensin/hour per ml (control=5+/-1), plasma aldosterone concentration to 99+/-23 pg/ml (control=14+/-4), and plasma sodium concentration to 142+/-1 mEq/liter (control=136+/-1). Urinary excretion of sodium decreased to 3.9+/-0.7 mEq/day and marked fluid retention occurred. We also found that these changes were accompanied by a decrease in hematocrit to 24+/-2% (control=40+/-1), formation of 36+/-9 ml of fluid in the thoracic cavity, 33+/-9 ml of ascites, pulmonary congestion and edema, hepatic congestion, and enlargement and hypertrophy of both the left and right ventricles. All rabbits died of ventricular failure at a time that was partly related to the degree of aortic constriction and that ranged from 2 to 12 days. The model we have established is chronic, highly reproducible, easy to produce, and inexpensive, and resembles the clinical syndrome of right and left congestive heart failure in man. Furthermore, the studies provide evidence for an important role of the renin-angiotensin-aldosterone system in the fluid retention that leads to pulmonary and systemic venous congestion after suprarenal aortic constriction.

Arterial pressure regulation during hemorrhage: homeostatic role of angiotensin II.

The role of the renin-angiotensin system in the maintenance of arterial pressure following hemorrhage was studied in conscious dogs. Hemorrhage (20 ml/kg body wt) decreased the mean arterial pressure, but compensatory mechanisms partially restored the arterial pressure toward normal. Plasma renin activity increased more than twofold following hemorrhage. To evaluate the role of endogenous angiotensin II in this compensatory response, a specific competitive antagonist of angiotensin II, 1-sarcosine-8-alanine-angiotensin II, was infused intravenously at 6.0 mug/kg min-1 for 30 min; the mean posthemorrhage arterial pressure decreased from 102 plus or minus 7 mmHg to 80 plus or minus 6 mmHg after 15 and 30 min of analog infusion (P less than 0.01 for both values). After a recovery period of 60 min, arterial pressure returned to pre-infusion levels. These results suggest that angiotensin II plays an important role in the short-term maintenance of arterial pressure following hemorrhage in the conscious animal.

Concentration of urine by the hibernating marmot.

Studies wer performed with marmots (Marmota flaviventris) of both sexes that had chronic arterial, venous, and bladder catheters. Urine collection was performed during hibernation and urine osmolalities (611.6 not equal to 166.1 SD) were found to be lower than those of aroused animals (1264 not equal to 472.9 SD), but hypertonic to plasma. Peak osmolality of meduallary slices was found to be in the range of osmotic pressures of urine obtained from hibernating or aroused animals. After single injections of a mixture of rho-aminohippurate and inulin, or during constant infusion of inulin, steady-state excretion by hibernators was not achieved for several days. Indirect evidence indicateds that the hibernating marmot is capable of PAH secretion.