Issue published January 1, 1973

T he outer, lateral esophageal walls in the distal half of the esophagus in each of five cats were labeled with small tantalum wires. About 8 wk later, esophageal motion associated with respiration and peristalsis, induced by injecting barium boli (5 ml each) into the proximal esophagus, was recorded on cine and serial biplane roentgenograms while recording intraluminal esophageal pressures simultaneously by manometry. Esophageal motion was also evaluated without a manometric tube in place. The coordinates for each marker were digitized and a computer was used to plot marker position against time. During respiration, the markers passively made a shallow, 2-10 mm excursion on the longitudinal esophageal axis. This movement was synchronous with thoracic and diaphragmatic movement and changes in intraluminal esophageal pressure. Immediately after the onset of peristalsis, the markers made a pronounced oral movement of 10 mm or more above their mean respiratory position, as if to engulf the bolus. Markers in opposing esophageal walls approximated one another and commenced an aboral movement as the bolus tail, which was essentially co-incident with onset of the manometric pressure complex, passed the marker sites. The markers returned to their respective rest positions essentially coincident with passage of the pressure complex peak and then moved below their respective rest positions. The aboral excursion occurred predominantly after the bolus had emptied into the stomach. The magnitude and duration of oral excursion was significantly greater for the distal than for the more proximal markers; conversely, the magnitude and duration of aboral excursion was greater for the proximal than for the more distal markers. During the peristaltic sequence, the labeled portion of the esophagus shortened from 26 to 46% of its resting length. No evidence of esophageal torque was shown. These findings suggest that both the longitudinal and circular esophageal musculature play an active and important role during peristaltic transport of a bolus through the esophagus.

T he purpose of this study was to determine the physiological mechanism of lower esophageal sphincter (LES) relaxation. Circular muscle of the esophagus, LES, and stomach were evaluated for their inhibitory response to electrical stimulation during a maintained tonic contraction produced by a superfusion of acetylcholine and physostigmine. Only the circular muscle of the distal esophagus showed an inhibitory response to electrical stimulation. The maximal inhibition of LES muscle was 63.9±5.9 (mean±SE)% of the acetylcholine produced tension and occurred at 80 V. Upper esophageal and gastric muscle were not inhibited. The inhibitory response of the LES muscle was antagonized by tetrodotoxin and hexamethonium but not by other specific antagonists. Adrenergic nerve destruction following 6-hydroxydopamine also did not abolish the LES inhibition. These data indicate that the distal esophagus, at the zone of the manometrically determined LES, is characterized by a nonadrenergic neural inhibitory system. We suggest that these nerves may mediate LES relaxation.

T he natriuresis of acute Ringer's loading is associated with a rise in the rate of delivery of fluid beyond the proximal tubule due both to a rise in glomerular filtration and a fall in absolute reabsorption, the latter being causally mediated, at least in part, by the accompanying fall in postglomerular vascular [protein]. To determine whether these factors also contribute to the renal response to chronic Ringer's loading, nine rats given continuous infusions, 30% body weight/day over 5-14 days, were studied using free-flow micro-puncture techniques. Results were compared with data from 10 chronic control rats given less than 1.5% body wt/day. Late proximal tubule fluid-to-plasma [inulin] ratios, (TF/P)_IN, single nephron glomerular filtration rate (SNGFR), absolute proximal reabsorption, and postglomerular vascular [protein] in chronic control rats and chronically loaded rats averaged 2.2±SE 0.1 (n = 35) and 1.5±0 (35), P<0.001; 37±2 (35) and 47±4 nl/min (35), P<0.05; 19±1 (35) and 16±2 nl/min (35), P>0.2; and 9.5±0.3 (8) and 8.6±0.3 g/100 ml (8), P>0.05, respectively. Thus the fall in (TF/P)_IN and the rise in distal delivery during chronic Ringer's loading were due almost entirely to the rise in SNGFR, and not to any large fall in absolute reabsorption. Hence chronic and acute Ringer's loading increase delivery of proximal tubule fluid by different mechanisms, with chronic sodium homeostasis being governed overwhelmingly by adjustments in GFR. When, however, an acute Ringer's load was infused into chronically loaded rats, we observed significant and parallel reductions in absolute proximal reabsorption and postglomerular vascular [protein]. These findings suggest that the difference between the effects of chronic vs. acute Ringer's loading on absolute proximal reabsorption may have been due, at least in part, to the corresponding difference in the effects these two loading procedures have on postglomerular vascular [protein].

A polipoproteins of the “C” group in human blood plasma, which contain the activator of the lipoprotein lipase-substrate interaction, were found to be transferred specifically from serum to phospholipid-stabilized fat emulsion. Content and distribution of apoprotein activator were measured in healthy men in the postabsorptive state and 4 h after ingestion of meals containing 100 g fat. Content of activator protein in whole serum did not change after ingestion of the fat-rich meals but that contained in triglyceride-rich lipoproteins of density (d) <1.006 approximately doubled whereas that of high density lipoproteins fell by half. The increased activator content of triglyceride-rich lipoproteins was virtually confined to chylomicrons and its concentration in chylomicron apoprotein was substantially greater than that in very low density lipoproteins. This difference could be ascribed largely to a higher content of C apoproteins in chylomicron protein since both the concentration of C apoproteins and of apoprotein activator were directly proportional to particle diameter while the pattern of fast-migrating C apoproteins in polyacrylamide gels was similar among chylomicrons and subfractions of very low density lipoproteins. Apparent concentration of activator protein was much greater in the high density lipoprotein subfraction of d 1.063-1.125 than in the subfraction of d 1.125-1.21. In the subfraction of d 1.063-1.125, the concentration of activator protein and of fast-migrating C apoproteins in polyacrylamide gels decreased after the fat-rich meal. Concentration of phospholipids in this fraction increased gradually to a peak 43% above the basal value 6 h after the meal. The results obtained demonstrate that high density lipoproteins contribute certain functionally important polar constituents to chylomicrons during alimentary lipemia in man and suggest that they also receive surface constituents from chylomicrons during the course of their metabolism.

S tudies were performed to define the mechanisms involved in the redistribution of renal cortical blood flow to inner cortical nephrons which occurs during hemorrhagic hypotension in the dog. The radioactive microsphere method was utilized to measure regional blood flow in the renal cortex. Renal nerve stimulation decreased renal blood flow 40% but had no effect on the fractional distribution of cortical blood flow. Pretreatment with phenoxybenzamine, phentolamine, propranolol, or atropine did not alter the redistribution of cortical flow during hemorrhage. A reduction in renal perfusion pressure by aortic constriction caused a qualitatively similar alteration in regional blood flow distribution as occurred during hemorrhage. When perfusion pressure was kept constant in one kidney by aortic constriction followed by hemorrhage, no redistribution occurred in the kidney with a constant perfusion pressure while the contralateral kidney with the normal perfusion pressure before hemorrhage had a marked increase in the fractional distribution of cortical flow to inner cortical nephrons. Additionally, retransfusion had no effect on the fractional distribution of flow in the kidney in which perfusion pressure was maintained at the same level as during hemorrhage while in the contralateral kidney in which pressure increased to normal there was a redistribution of flow to outer cortical nephrons. These studies indicate that the redistribution of renal cortical blood flow which occurs during hemorrhage is not related to changes in adrenergic activity but rather to the intrarenal alterations which attend a diminution in perfusion pressure.

I n an effort to better define the role of βadrenergic blockade in human bronchial asthma, peripheral blood leukocytes and lymphocytes from individuals with this condition were studied for possible alterations in cyclic AMP metabolism. Using a previously described radioimmunoassay to measure cyclic AMP, cells from asthmatic subjects were shown to have a highly significant decrease in their cyclic AMP response to β-adrenergic agents (isoproterenol, norepinephrine, and epinephrine) by comparison with normal control cells. The alteration in responsiveness was most marked at the time of severe active asthma and returned toward normal during periods of clinical remission. Evidence was presented to indicate that the reduced response in cells from asthmatic individuals was not due to marked alterations in the proportion of T and B lymphocytes. Five normal volunteers were treated with an oral bronchodilator preparation containing theophylline and ephedrine over a 2 wk period without a significant change in the lymphocyte cyclic AMP response.These results provide unambiguous evidence for altered adrenergic responsiveness in bronchial asthma and indicate that purified peripheral blood lymphocytes should be a suitable in vitro system for further elucidation of the abnormality.Despite the reduction in catecholamine responsiveness in the asthmatic population as a whole, major alterations were largely restricted to individuals with severe, chronic asthma. Conclusive evidence for β-adrenergic blockade in individuals who have not had recent asthmatic symptoms was not obtained, casting some doubt on the theory that bronchial asthma is due to a congenital derangement of cyclic AMP metabolism. Moreover, transient episodes of bronchospasm were often accompanied by a normal cyclic AMP response indicating that episodes of asthma frequently occur in the absence of easily demonstrable adrenergic blockade.

T hyroxine (T₄) and triiodothyronine (T₉) are rapidly degraded by a purified preparation of myeloperoxidase (MPO) and H₂O₂ with the formation of iodide and material which remains at the origin on paper chromatography. Deiodination by MPO and H₂O₂ occurs more readily at pH 7.0 than at pH 5.0 in contrast to iodination by this system which is known to occur more readily at pH 5.0 than at pH 7.0. Degradation is inhibited by azide, cyanide, ascorbic acid, and propylthiouracil. Methimazole stimulates deiodination by MPO and H₂O₂ but inhibits this reaction when MPO is replaced by lactoperoxidase or horseradish peroxidase.Intact human leukocytes, in the resting state, degrade T₄ and T₃ slowly: degradation, however, is increased markedly during phagocytosis of preopsonized particles. Serum inhibits this reaction. T₃ can be detected as a minor product of T₄ degradation. Proteolytic digestion of the reaction products increases the recovery of monoiodotyrosine. The fixation of iodine in the cytoplasm of leukocytes which contain ingested bacteria was detected radioautographically. Chronic granulomatous disease leukocytes, which are deficient in H₂O₂ formation, degrade T₄ and T₃ poorly during phagocytosis. MPO-deficient leukocytes degrade the thyroid hormones at a slower rate than do normal leukocytes although considerable degradation is still observed. Azide, cyanide, ascorbic acid, and propylthiouracil which inhibit certain peroxidasecatalyzed reactions inhibit degradation by normal leukocytes; however, inhibition is incomplete. Formation of iodinated origin material is inhibited to a greater degree by azide, cyanide, and propylthiouracil than is deiodination. Methimazole inhibits the formation of iodinated origin material by both normal and MPO-deficient leukocytes. However, deiodination by normal leukocytes is stimulated and that of MPO-deficient leukocytes is unaffected by methimazole. Hypoxia inhibits the degradation of T₄ and T₃ by untreated normal or MPO-deficient leukocytes and by normal leukocytes treated with azide or methimazole.These data suggest that both MPO-dependent and MPO-independent systems are involved in the degradation of T₄ and T₃ by phagocytosing leukocytes. The role of leukocytic degradation of T₄ and T₃ in thyroid hormone economy and in leukocytic microbicidal activity is considered.

S pecific quantitative techniques have been used to measure the cytoplasmic estradiol-binding protein (EBP) in human mammary carcinoma tissue specimens. Sucrose gradient centrifugation reveals EBP to sediment at 8S and 4S. Variable quantities of non-specific estradiol binding occurs in the 4S region of the sucrose gradient necessitating controls to insure specificity of the estradiol protein interaction.Using dextran-coated charcoal to separate bound from free estradiol Scatchard analysis finds the dissociation constant of the estradiol EBP interaction to be ∼ 2.6×10^-10 M, indicative of the very high affinity of the ligand for the EBP. Quantitation of EBP sites in 64 primary and metastatic human breast tumors demonstrates a continuous spectrum of values from 0 to 612 fmol per mg of cytoplasmic protein. Specific 8S binding in the sucrose gradient centrifugation was not detected in specimens containing less than 9.0 fmol EBP per mg cytoplasmic protein.Since data from animal breast tumors and preliminary evidence from human breast tumors indicates an excellent correlation between the presence of abundant tumor EBP and endocrine-induced breast cancer regressions, precise quantitation of EBP in all human primary tumors may prove to be an excellent prognosticator of endocrine therapy in metastatic breast cancer.

T he effects of acute Salmonella typhimurium sepsis on the kinetics of peripheral L-thyroxine (T₄) distribution and metabolism and on serum total and free T₄ concentrations were studied in rhesus monkeys inoculated i.v. with either heat-killed or viable organisms. The rate of disappearance of labeled T₄ from serum was increased within 8 h after inoculation of monkeys with either heat-killed or viable Salmonella. The effects of the heat-killed organisms were transient and no longer evident by 16 h postinoculation. The monkeys inoculated with the viable Salmonella experienced a 2-3 day febrile, septic illness that was accompanied by an increase in the absolute rate of T₄ disposal. In the infected monkeys, serum total T₄ and endogenously labeled protein-bound iodine concentrations fell significantly during the period of acute sepsis and then rose during convalescence to values that exceeded the preinoculation values, suggesting that thyroidal secretion of hormone had increased in response to a primary depletion of the peripheral hormonal pool. Total cellular and hepatic uptakes of T₄ were enhanced by 4 h after inoculation of monkeys with either heat-killed or viable Salmonella, but the increase in total cellular uptake persisted for 24 h only in the monkeys inoculated with the viable organisms. These alterations in T₄ kinetics could neither be correlated with changes in the binding of T₄ in plasma nor attributed to an increase in vascular permeability. Moreover, they could not be ascribed to an in vitro product of bacterial growth, suggesting that the presence of the organisms themselves was required. An acceleration of T₄ disappearance was also observed during Escherichia coli and Diplococcus pucumoniae bacteremias. Our findings are consistent with a primary increase in the cellular uptake and metabolism of T₄ during bacterial sepsis, possibly related to phagocytic cell function in the host.

T he serous glands of rat tongue were found to contain a potent lipolytic enzyme which hydrolyzed triglyceride to mostly diglyceride and free fatty acids (FFA) at pH 4.5-5.4. Homogenates of lingual serous glands from adult rats hydrolyzed 40-70 mmol of triglyceride/g per h. The soft palate, anterior oral pharyngeal wall, and lateral oral pharyngeal glands also contained the activity, but at a much lower level. The lipolytic activity was also found in saliva collected through an esophageal cannula and in stomach contents of rats fed a fat-rich meal. The stomach contained very little activity, however, when saliva was excluded. Lipolytic activity was not found in the stomach wall or in the parotid, submandibular, and sublingual glands. The findings suggest that the lingual serous glands secrete a lipase which catalyzes in the stomach the conversion of triglyceride to partial glycerides and FFA. It is proposed that this reaction is the first step in the digestion of dietary lipid.

U roporphyrin I is found in high concentration in the bones, teeth, blood, soft tissues, and urine of the fox squirrel, Sciurus niger. The concentration of uroporphyrin in fox squirrel spleen is much higher than in liver, kidney or bone marrow, probably because of accumulation from phagocytosed red cells. Bleeding causes a marked increase in the uroporphyrin concentration of red cells and spleen, and a 3-8-fold increase in uroporphyrin excretion. Urinary excretion of δ-aminolevulinic acid and porphobilinogen is not greater in fox squirrels than in nonporphyric gray squirrels. Sciurus carolinensis, used as controls. In all these characteristics, as well as in the previously demonstrated deficiency of the enzyme uroporphyrinogen III cosynthetase in red cells, the physiological porphyria of fox squirrels resembles congenital erythropoietic porphyria, a hereditary disease of man and cattle. For squirrels differ in showing no evidence of cutaneous photosensitivity or hemolytic anemia.Uroporphyrinogen III cosynthetase activity is present in fox squirrel bone marrow at 1/10 its concentration in gray squirrel marrow. The fox squirrel enzyme is much more unstable than the gray squirrel enzyme, which provides a possible explanation for its low activity and for the overproduction of uroporphyrin I. It is unlikely that the deficiency of cosynthetase is due to its inactivation by excessive amounts of uroporphyrinogen I synthetase, because activity of the latter enzyme is the same in blood from fox and gray squirrels.Fox squirrel porphyria provides a convenient model for studies of pathogenesis of human congenital erythropoietic porphyria.

T he problem as to whether iodohistidines are normally biosynthetized in thyroglobulin and thyralbumin has been examined both in man and the rat. Evidence has been obtained for the first time that diiodohistidine (DIH) is present in both species in these two iodoproteins. The biosynthesis of monoiodohistidine (MIH) in the thyroglobulin of the normal rat has been confirmed and extended to rat thyralbumin and to human thyroid iodoproteins.The iodohistidine identification is based on five original methods including: (a) the preparation of stable and radioiodine-labeled iodohistidines; (b) the protection of the labile iodohistidines during the iodoprotein enzymatic hydrolysis; (c) the isolation of iodohistidines by ion-exchange resin chromatography; (d) their separation from each other and from iodinated cationic butanol-insoluble compounds by Sephadex G-10 chromatography; and (e) their purification by successive crystallizations to a constant specific activity.Iodohistidine levels (in percent of protein radioactivity from iodide given in vivo) were found comparable in man and the rat. However, the values (mean ±SE) for thyroglobulin (MIH, 0.61±0.10%; DIH, 0.050±0.015%) and for thyralbumin (MIH, 2.61±0.57%; DIH, 0.28±0.09%) differ significantly (P < 0.05).Iodohistidines are stable during in vitro exposure to iodotyrosine dehalogenase preparations. In contrast to iodotyrosines the iodohistidines when given in vivo to man either orally or intravenously were in large part recovered in 24-h urines.

B utanol-insoluble iodinated compounds in the urine of patients with congenital goiters have been generally regarded as iodopeptides. Monoiodohistidine (MIH) and diiodohistidine (DIH) were identified from the urine of four patients with congenital goitrous hypothyroidism. From radioiodine studies, 40-70% of the urinary radioactivity was in the iodide-free fraction from which about 40% was identified as MIH and DIH by crystallizations to a constant specific activity.Iodotyrosines were simultaneously identified in the urine. However the presence of an iodotyrosine-deiodinase activity was demonstrated in the two removed goiters with a normal K_m for MIT. In vivo iodotyrosine deiodination was normal for hypothyroid subjects.No thyroglobulin was identified in the thyroids from these patients. The major iodoprotein was iodoalbumin which, after in vivo labeling, contained 84-89% of the total soluble protein radioactivity. The thyroxine content of the goiter iodoalbumins and other iodoproteins was extremely low.Iodohistidines were identified in comparable proportions in the iodoalbumin and in the other iodoproteins isolated from each goiter. The average iodohistidine content of these proteins as crystallizable MIH and DIH was in the individual cases 15 and 4% of the in vivo incorporated radioiodine. DIH was identified in all iodoprotein fractions. The mean DIH/MIH ratios from the individual cases were 1.16 and 0.35. The corresponding DIT/MIT ratios were 3.19 and 1.45, respectively.The major consequence of this thyroglobulin defect is the iodination of inappropriate proteins (mainly albumin) resulting in low yields of thyroxine and high yields of iodohistidines. Iodohistidines from the goiter iodoproteins were not deiodinated and, at least for MIH, were quantitatively excreted in the urine of these patients. From the MIH iodoalbumin content and the MIH urinary excretion, goiter iodoalbumin turnover estimates were made and, although elevated, could not maintain a normal thyroxine secretion.The urinary excretion of iodohistidines easily demonstrated by column chromatography is offered as a test for detecting this variety of congenital goiter.

T he leukocytes of 16 adult patients with acute myeloblastic leukemia were studied by autoradiographic methods to elucidate the mode of action of daunomycin.It was shown that daunomycin, at clinically useful doses, exhibits a cytolytic effect on all leukemic blasts whatever their cell-cycle phase. This cytolytic action affects, however, preferentially S-phase cells.It was shown also that blasts of patients less sensitive to daunomycin or receiving a lesser dose of the drug are temporarily blocked in G₂ phase (delayed mitosis) or in G₂ phase (prolonged generation time).Finally daunomycin appeared to hamper the passage of G₂-blocked blasts from the bone marrow to the blood, while G₂-phase cells crossed freely.

C irculating levels of immunoreactive parathyroid hormone (PTH) were measured in 40 patients with idiopathic hypercalciuria (IH) before and during reversal of hypercalciuria with thiazide, and in four normal subjects before and during induction of hypercalciuria with furosemide. 26 patients with IH had elevated serum PTH levels. The remaining patients had normal levels. Although the correlation was not complete, high PTH levels were generally found in patients who had more severe average urinary calcium losses. When initially elevated. PTH levels fell to normal or nearly normal values during periods of thiazide administration lasting up to 22 months. When initially normal, PTH levels were not altered by thiazide. Reversal of hyperparathyroidism by thiazide could not be ascribed to the induction of hypercalcemia, since serum calcium concentration failed to rise in a majority of patients. Renal hypercalciuria produced by furosemide administration elevated serum PTH to levels equivalent to those observed in patients with IH.The findings in this study help to distinguish between several current alternative views of IH and its relationship to hyperparathyroidism. Alimentary calcium hyperabsorption cannot be the major cause of IH with high PTH levels, because this mechanism could not elevate PTH. Idiopathic hypercalciuria cannot be a variety of primary hyperparathyroidism, as this disease is usually defined, because PTH levels are not elevated in all patients and, when high, are lowered by reversal of hypercalciuria. Primary renal loss of calcium could explain the variable occurrence of reversible hyperparathyroidism in IH, since renal hypercalciuria from furosemide elevates serum PTH in normal subjects. Consequently, a reasonable working hypothesis is that IH is often due to a primary renal defect of calcium handling that leads, by unknown pathways, to secondary hyperparathyroidism.

I t has been confirmed that the rabbit vermiform appendix secretes spontaneously at a relatively rapid rate (1-12 ml·h^-1; 1.4±0.24 μl·min^-1·cm^-2). The electrolyte composition is similar to that of ileal fluids and independent of the secretory rate. The transmural potential difference is about 12 mV, mucosa negative. Of the major electrolytes, only HCO₃^- is secreted grossly against its electrochemical potential difference. This finding plus the low hydraulic (or osmotic) permeability (L_p) and high secretory pressures of the organ strongly suggest that the secretion is an active one. The passive permeability to Na⁺ and Cl^- appears to be, at most, somewhat less than for small bowel. Permeability to mannitol was estimated at 2.5 × 10^-7 cm·s^-1. On the basis of reasonable assumptions and results with luminal test solutions of differing osmolarities, it was concluded that (a) the L_p of the appendiceal epithelium is in the lower range of values reported for small bowel and colon; (b) the L_p is higher for osmotic absorption than for osmotic secretion; and (c) the rate of spontaneous secretion is insensitive to luminal anisotonicity over a wide range of values. But sufficiently hypotonic solutions can reverse net secretion to net absorption, more by inhibiting spontaneous secretion than increasing osmotic absorption. The rabbit vermiform appendix appears to be a useful model for the elucidation of intestinal secretory processes.

H ypocalcemia is a frequent feature of hypomagnesemia in man and several other species. To elucidate the cause of this hypocalcemia, we have studied a child with primary hypomagnesemia and secondary hypocalcemia during magnesium supplementation when he was normomagnesemic and normocalcemic and after magnesium restriction for 16 days when he quickly became hypomagnesemic (0.5 meq/liter) and hypocalcemic (3.4 meq/liter) and had positive Chvostek's and Trousseau's signs.Whether in the normomagnesemic or hypomagnesemic state, intravenous bovine parathyroid extract (PTE) 8 U. S. P. U/kg promptly caused transient increases in the urinary phosphate excretion, renal phosphate clearance and cyclic AMP excretion. The magnitudes of these responses were similar in the two states, and similar to those observed in a hypoparathyroid patient. When the patient was hypomagnesemic and hypocalcemic, intramuscular PTE, 8 U/kg at 8-h intervals for four doses promptly caused hypercalcemia. The findings indicate that the end-organs were responsive to parathyroid hormone.The concentrations of serum parathyroid hormone (PTH) were normal in the normomagnesemic state ranging from 0.15 ng/ml to 0.40 ng/ml. Serum PTH did not increase in the hypomagnesemic state in spite of hypocalcemia. Indeed, PTH became unmeasurable in four consecutive samples at the end of the period of magnesium restriction.The concentrations of serum calcitonin remained unmeasurable (< 0.10 ng/ml) throughout the study, implying that excess calcitonin was not the cause of hypocalcemia in magnesium depletion.The findings in this study support our thesis that magnesium depletion causes impaired synthesis or secretion of parathyroid hormone. This impairment would account for the hypocalcemia observed in the hypomagnesemic state.

P henobarbital, by inducing liver microsomal enzymes, may affect bile acid synthesis from cholesterol and thus alter the secretion of biliary lipids and the composition of bile. We, therefore, determined the effects of phenobarbital on bile flow, biliary lipid secretion, bile acid synthesis, and bile-acid pool size. Using an experimental preparation that allows controlled interruption of the enterohepatic circulation (1), we administered 5 mg/kg per day of phenobarbital to healthy Rhesus monkeys for 1-2 wk to achieve steady-state conditions. Three animals were studied with an intact enterohepatic circulation and three with a total bile fistula, each animal served as its own control. Total bile flow and secretion of bile salt, phospholipid, and cholesterol were measured every 24 h during steady-state conditions. Further, under conditions of an intact enterohepatic circulation bile-acid synthetic rate was measured in three animals and pool size estimated in two animals during both control and drug treatment periods.Phenobarbital at doses of 5 mg/kg per day increased bile flow 30-50% in all animals (P < 0.001). The increased bile flow resulted from both an increased “bile-salt independent fraction” and an increased bile-salt secretion rate. Phenobarbital significantly increased bile salt (P < 0.01) and phospholipid secretion (P < 0.05) by about 30% but cholesterol secretion was not significantly changed. Consequently, the concentration of cholesterol relative to bile salt and phospholipid was decreased (P < 0.001). Phenobarbital significantly enhanced the maximal rate of bile acid synthesis 25-30% in all three monkeys with total bile fistulas (P < 0.05) and also augmented bile acid synthesis and pool size in animals with intact enterohepatic circulations despite the fact that the rates of bile salt returning to the liver in these animals would have inhibited bile acid synthesis in control animals. Thus, phenobarbital not only increases the maximal rate of bile acid synthesis but also alters the normal control mechanisms by which bile salts returning to the liver inhibit bile salt synthesis. The fact that phenobarbital treatment results in increased synthesis of bile salt and unchanged secretion of cholesterol is consistant with the view that the drug augments conversion of hepatic cholesterol to bile salt. The resulting decrease in relative cholesterol content in bile may have therapeutic implications for cholesterol gallstone therapy.

P arathyroid function was assessed by calcium infusions (4-8 h) in 16 patients with chronic renal insufficiency being treated by long-term hemodialysis. The concentrations of two immunoreactive species of parathyroid hormone in plasma (iPTH-9, mol wt 9500; iPTH-7, mol wt 7000) were estimated by radioimmunoassays utilizing two relatively specific antisera. Control values of the smaller species, iPTH-7, were uniformly high, whereas values of iPTH-9 were normal in 12 of 19 studies. Response of iPTH-7 to calcium infusions was variable, with significant decreases occurring only five times in 27 infusions. Concentrations of iPTH-9, however, decreased during every calcium infusion. In contrast to these acute responses, five of six patients studied during periods of dialysis against both low (< 6 mg/100 ml) and high (7-8 mg/100 ml) calcium concentrations in the dialyzate showed a decrease in values of iPTH-7 during the period of dialysis against the higher calcium concentration. It is concluded that plasma concentrations of iPTH-9 reflect primarily the moment-to-moment secretory status of the parathyroid glands, while concentrations of iPTH-7 reflect more closely chronic parathyroid functional status. It is further concluded that the failure of iPTH-7 to decrease during induced hypercalcemia should not be equated with autonomy of parathyroid gland function.

T wo major species of serum immunoreactive parathyroid hormone (iPTH) were measured in 47 untreated patients with primary osteoporosis by using two highly specific radioimmunoassays. Mean iPTH was normal with one antiserum but was lower than normal (P < 0.001) with the other, iPTH values did not correlate with biochemical parameters or with the proportion of bone-resorbing surfaces in iliac crest bone biopsy specimens. These data suggest that the increased bone resorption is not due to increased parathyroid function in most osteoporotic patients. However, seven of our patients (15%) appear to represent a separate population because they had increased values with one or the other of the antisera.

U rinary hemibladders obtained from toads soaked in water or saline were treated with aldosterone, 10^-6 M, either 1½ or 16 h after mounting. After 2½ h exposure to the hormone, short-circuit current was increased by 110-192% and open-circuit potential by 20-44% as compared with untreated paired hemibladders. Mucosal cells were then assayed for sodium-potassium-stimulated adenosine triphosphatase (ATPase). No increase occurred in activity per milligram protein or in the portion of total activity dependent on sodium. Activity at low sodium concentrations was also measured and analyzed by means of the Hill equation in terms of K, the apparent dissociation constant of the enzyme-sodium complex, and n, a number that expresses the degree of interaction between binding sites. Neither K nor n was significantly altered by aldosterone. A few experiments were also carried out at low ATP concentrations (0.3 mM); again no change in sodium-dependent activity was noted. The results indicate that aldosterone does not stimulate sodium transport by increasing the quantity of sodium-potassium adenosine triphosphatase in mucosal cells or the dependence of this activity on sodium or ATP concentrations.

I n recent studies in this laboratory employing normal hydropenic rats we have demonstrated that the reduction in absolute proximal reabsorption that attends the experimental reduction of single nephron glomerular filtration rate (SNGFR) (glomerulotubular balance) is mediated, at least in part, by the accompanying decline in postglomerular vascular protein concentration, and therefore, postglomerular colloid osmotic pressure (πEA). The present study was undertaken to define the quantitative contribution of these changes in πEA to the changes in absolute proximal reabsorption measured under these conditions. A protocol was employed which enabled us to examine the effects on absolute proximal reabsorption of reductions in filtered load brought about under conditions in which πEA remained essentially unchanged. Thus, after partial aortic constriction in 16 plasma-loaded rats, near constancy of πEA was observed in 10 (a change in efferent arteriolar protein concentration of 0.4 g/100 ml or less) and in these, uniform reductions in SNGFR averaging 16.7 nl/min were attended by reductions in absolute proximal reabsorption averaging only 1.7 nl/min, or 7% of preconstriction values. These findings, taken together with previous observations from this laboratory, suggest that the proximal reabsorptive adjustment that characterizes glomerulotubular balance in the rat is markedly blunted when changes in πEA are prevented. In the remaining six rats, a mean reduction in filtered load comparable to that observed in the above group was attended by slightly to moderately greater reductions in efferent arteriolar protein concentration, thereby fulfilling less well the stated aim of this study. Nevertheless, in accord with the above conclusion, these relatively greater reductions in πEA were accompanied by correspondingly greater reductions in absolute proximal reabsorption.

S ingle-stranded DNA (SDNA) occurs in high incidence and in greatest concentration in the sera of patients with systemic lupus erythematosus (SLE), where levels as high as 250 μg/ml were observed. SDNA appears to be an imunogen for anti-SDNA antibodies and forms complexes in vivo of both anti-SDNA-SDNA and anti-NDNA-SDNA types, which apparently play a role in the pathogenesis of the glomerulonephritis found in patients with SLE, SDNA is also found in high incidence but at lower levels in the sera of patients with rheumatoid arthritis. Lesser amounts of SDNA are found in several other diseases in which a low incidence of anti-SDNA antibodies is observed.

T he relative roles of triiodothyronine (T₃) and thyroxine (T₄) in modulating pituitary responsiveness to thyrotropin-releasing hormone (TRH) have been assessed. (a) 10 hyperthyroid patients with elevated serum T₂ and T₄ levels showed no pituitary response to TRH. After 2 wk of propylthiouracil therapy T₄ levels had fallen to normal in only five patients while T₂ levels were normal in all. Pituitary responsiveness to TRH returned in all patients with normal or high T₄ concentrations. (b) Patients with isolated elevations of serum T₃ (T₃ toxicosis) failed to respond to TRH. TRH responsiveness was restored when T₃ levels fell to normal after propylthiouracil therapy. (c) When pituitary responsiveness to TRH was tested 60 min after a single oral dose of 50 μg of T₃, which increased serum T₃ levels to slightly above the normal range, no rise in thyrotropin (TSH) was seen in six subjects. These findings indicate that T₃ elevations alone can rapidly inhibit pituitary responsiveness to TRH.

T he effects of stimulation of the mixed autonomic nerve to the dog pancreas has been studied under conditions in which both pancreaticoduodenal vein blood flow and insulin concentration were determined. Stimulation resulted in increased insulin output, which was blocked by prior administration of atropine. Blood flow was reduced by stimulation in proportion to the rate of stimulation. At 40 stimuli/s a maximum effect was found at 1 min with a gradual return toward base line despite continued application of the stimulus. Atropinization had no effect on blood flow changes. Insulin responses to 0.1 g/kg glucose were reduced on the average 40% by simultaneous stimulation of the pancreatic nerve at 40 cycles/s in atropinized animals. These studies establish this preparation as a reproducible model for the direct examination of autonomic influences on endocrine pancreatic function. From them it is concluded that the nerve supply to the endocrine pancreas of the dog is sufficient to inhibit insulin secretion by activation of the sympathetic nerves and to stimulate insulin secretion by activation of the parasympathetic nerves.

D eficiency of cystathionine synthase activity results in the clinical syndrome of homocystinuria. Using phytohemagglutinin (PHA)-stimulated lymphocytes as a readily available source of this enzyme, its activity has been compared in 48 control subjects, seven homozygotes affected with homocystinuria, and 17 obligate heterozygotes. PHA-induced enzyme levels were highest in controls (mean ±SEM, 666.9±70.2 pmol cystathionine formed/mg protein per 4 h), intermediate in heterozygotes (114.4±27.3), and absent to severely deficient in homozygotes (2.0±1.6). Since only three of the 17 values from the obligate heterozygotes overlapped into the control range, this simple method may become clinically useful for heterozygote detection of carriers of the gene for abnormal cystathionine synthase. In addition, this system for induction of cystathionine synthase in lymphocytes has a more general relevance to human biochemical genetics in that it demonstrates that the absence of an enzyme in a normal cell does not preclude using that source for diagnosis of genetic disease if the enzyme can be induced.