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Temporary induction by ethanol of normal glycogenolysis in patients with liver glycogen disease
Volume 67
Number 5
part 2
challenge with RSSE virus. In contrast, pools of 7G-immunoglobulins, containing neutralizing antibody, afforded significant passive protection. The difference in protective activity is probably related to differences in physicochemical structure of the two immunoglobulins.
60. Coproantibodies to bacterial and viral enteropathogens in breast-fed infants Jean F. Kenny,* Mary I. Boesman,* and Richard H. Micheals,* Children's Hospital and University of Pittsburgh, Pittsburgh, Pa. Introduced by Richard L. Day The purpose of this study was to determine whether significant amounts of antibody in h u m a n milk survive passage through the intestinal tract. Schubert and Grfinberg (Schweiz. reed. Wchnschr. 79: 1007, 1949) found typhoid agglutinins in stools of infants fed high titer colostrum. Hodes (]. Pediat. 65: 1017, 1964) reported that at least some of the colostrum beta-2A globulin with antibacterial and antiviral activity appears unchanged in the infant's stool. The present study revealed that stools of newborn breast-fed infants may contain surprisingly large amounts of hemagglutinating antibodies to three types of enteropathogenic E. ¢oli and also neutralizing antibodies to type 1 poliovirus. Antibody levels were measured in milk, infant and maternal sera, and supernatants from 20 per cent suspensions of stool. Newborn stool antibody titers were usually two- to eightfold lower than those in milk, averaging fourfold lower for antibacterial and only twofold lower for antiviral activity (in the one infant tested at 5 weeks, there was still only a twofold difference): Stool antibody levels correlated well with those of milk, but poorly with those of infant and maternal sera. Coproantibodies were not found in bottle-fed infants, except for traces of antiviral activity in meconium when infant serum antibody titers were high. Antibodies in milk and stool were specific and relatively resistant to trypsin. Bacterial hemagglutinins were sensitive to mercaptoethanol, whereas the viral neutralizing activity was not. Stool extracts were concentrated tenfold for immunodiffusion studies. Transitional and later stools from breast-fed infants were found to contain gamma-l-globulins similar to those in milk, including IgA and small amounts of IgM. No h u m a n immune globulins were detected in stools from artificially fed infants.
61. The etiology of acquired hemolytic anemia
Abstracts
94 3
Wolf W. Zuelzer, Cyril S. Stulberg, Robert H. Page,* Jos~ Teruva,* and A. Joseph Brough,* Child Research Center of Michigan, Children's Hospital of Michigan, and Wayne State University School of Medicine, Detroit, Mich. Clinical, pathologic, and virologic observations in children with acquired hemolytic anemia (AHA) will be presented, indicating the consistent association of hemolysis, irrespective of the presence of autoimmune phenomena, with active disseminated viral infection attributable to the agent of eytomegalic inclusion disease ( C I D ) . The extent of viral activity in the R.E. system became apparent when a morphologic study of the evolution of the classical cellular lesion disclosed recognizable early and intermediate stages of infection at the cellular level and thus led to the demonstration of vast numbers of virus-infected cells in lymph nodes and other tissues at time of active hemolysis. Acute transient AHA was observed chiefly in association with primary CID in otherwise normal infants showing the classical syndrome of generalized infection, hepatitis, or a theretofore unrecognized milder form manifested by generalized lymphadenopathy, hemolysis, and at times, a characteristic rash. The virus as an agent with latency and low virulence meets the theoretical requirements for an etiologic agent sustaining the chronic, intermittent course of AHA. In a high percentage of subjects with chronic intermittent AHA, evidence of impairment of immune mechanism was found, which seemed to account for the persistent or recurrent viral activity, on the basis of a disturbed hostvirus equilibrium. These observations led to the conclusion, supported by serologic-hematologic findings, that autoantibodies in AlIA represent a variable secondary response to viral infection in the R.E. system.
METABOLISM
62. Temporary induction by ethanol of normal glycogenolysis in patients with liver glycogen disease Charles U. Lowe, Luis L. Mosovich,* and Valiollah Abbassi,* State University of New York at Buffalo, Buffalo, N. Y. Observations made during alcohol infusion in 7 patients with hyperlacticemia and hypoglycemia due to liver glycogen disease ( L G D ) challenge the causal relation between enzyme defects observed in vitro and metabolic abnormalities in the whole patient. Oxidation of E t O l i to acetaldehyde is catalyzed by alcohol dehydrogenase with reduction of D P N to D P N H . In a coupled reaction, pyruvate is converted to lactate with regeneration of DPN. When alcohol is consumed
9 44
Society for Pediatric Research
after a fast to remove liver glycogen, lactate and FFA levels in blood rise; the D P N H produced inhibits the conversion of UDP-gal to UDP-glu and glutamate to alpha ketoglutarate. Galactose removal from blood is markedly delayed and gluconeogenesis from amino acids is inhibited. I n contrast with predictions, when acohol was infused into these 7 patients, no hypoglycemia developed; lactate and pyruvate levels fell to normal; FFA failed to rise; alcohol levels ranged from 0 to 22 rag. per cent (controls 90-120 mg. per cent) and there was no interference with galactose removal. In addition, their abnormal hyperglycemic response to glucagon was now entirely normal. These data suggest that alternate pathways for glyeogenolysis as well as alcohol oxidation should be sought; in addition, they challenge conventional thinking in relation to LGD, raising questions as to the significance of observed abnormalities in glucose-6-phosphatase activity and recommend the use of caution in assigning a causal role to abnormalities in enzyme function demonstrated by in vitro studies.
63. Regulation of pyruvate metabolism during development Everett Lovrien,~ Gerald Reaven, ~" and Robert E. Greenberg, Stanford University School of Medicine, Palo Alto, Calif. The importance of various mechanisms controlling formation of glucose from pyruvate is not well defined. As a means of delineating the role of some factors controlling gluconeogenesis during development, the effects of age, fasting, and administration of hydrocortisone on the metabolism of pyruvate by slices of kidney and liver were investigated. Two approaches were used: (1) incorporation of C 14 from pyruvate2-C 1~ into glucose, lipid, glycogen, and CO2, and (2) measurement of activities of pyruvate carboxylase (pyr-C) and phosphoenolpyruvate carboxykinase (PEP-C), which catalyze the first step in gluconeogenesis, conversion of pyruvate to phosphoenolpyruvate. Conversion of pyruvate-C 14 to glucose-C 14 was minimal in slices of fetal kidney and liver, whereas incorporation of (314 into lipid was greater in both tissues during fetal life. Activity of pyr-C and PEP-C was low in both tissues during fetal life, increasing with age. Gluconeogenesis increased in liver slices from newborn and weakling rats following a fast and hydrocortisone; activity of PEP-C similarly increased, while that of pyr-C did not change. Ilydrocortisone increased pyruvate conversion to glucose in kidney slices only from weanling rats, while fasting had no effect at any age. Activity of pyr-C increased with hydrocortisone and fasting only in kidney slices from newborn and weanling rats, respectively, while activity of PEP-C increased following hydrocortisone and fasting in kidney from both age groups. These studies reflect the complexity of mechanisms c o n -
November 1965
trolling gluconeogenesis and indicate that conversion of pyruvate to phosphoenolpyruvate is not the only rate-limiting step in gluconeogenesis during development.
64. Renal phosphate clearance and bicarbonate excretion in the ]etal lamb Fred G. Smith, Jr., Birger O. Tinglof,* and Forrest H. Adams, University of California School of Medicine, Los Angeles, Calif. Neonatal tetany with associated hyperphosphatemia may be related to transitory hypoparathyroidism, renal insensitivity to parathormone, or both. Twelve lambs near term were delivered by cesarean section with special precaution to maintain an intact placental circulation. The fetalmaternal preparation was monitored by measuring arterial pressures, blood pH, pCO2, and body temperatures. Fetal inulin, phosphate, and calcium clearances were measured before and after the intravenous administration of parathyroid extract (0.1 unit per minute per kilogram). In all experiments the clearance of phosphate was calculated as the ratio of the observed clearance to that of inulin. In all fetuses the control phosphate clearances were extremely low (0 to 0.02 c.c. per minute per kilogram) in spite of high serum phosphorus concentrations (mean 9.9 mg. per 100 c.c.). Following parathyroid extract the phosphate clearances ( C p / C I , x 100) changed significantly (0.03 to 0.09 c.c. per minute per kilogram, P < 0.01) demonstrating that the fetal kidney does respond to parathyroid extract. Bicarbonate excretion rates were also studied in 5 fetuses before and after acetazolamide. Bicarbonate excretion increased rapidly from a mean of 1.8 #Eq to 16.1 ~Eq per minute following enzyme inhibition suggesting an active carbonic anhydrase mechanism in the fetal kidney. These studies demonstrate that fetal hyperphosphatemia is due to factors other than insensitivity of the renal tubule to parathormone.
65. Familial heterotopic calcification and byperpbospbatemia unresponsive to parathyroid extract Warren F. Dodge, ¢ Luther B. Travis, ~ and Manuchehr Assemi,* University of Texas Medical Branch, Galveston, Texas Introduced by C. William Daeschner T h e reported cases of heterotopic calcification, accompanied by hyperphosphatemia, normocalcemia, and absence of renal insufficiency, are few.
Number 5
part 2
challenge with RSSE virus. In contrast, pools of 7G-immunoglobulins, containing neutralizing antibody, afforded significant passive protection. The difference in protective activity is probably related to differences in physicochemical structure of the two immunoglobulins.
60. Coproantibodies to bacterial and viral enteropathogens in breast-fed infants Jean F. Kenny,* Mary I. Boesman,* and Richard H. Micheals,* Children's Hospital and University of Pittsburgh, Pittsburgh, Pa. Introduced by Richard L. Day The purpose of this study was to determine whether significant amounts of antibody in h u m a n milk survive passage through the intestinal tract. Schubert and Grfinberg (Schweiz. reed. Wchnschr. 79: 1007, 1949) found typhoid agglutinins in stools of infants fed high titer colostrum. Hodes (]. Pediat. 65: 1017, 1964) reported that at least some of the colostrum beta-2A globulin with antibacterial and antiviral activity appears unchanged in the infant's stool. The present study revealed that stools of newborn breast-fed infants may contain surprisingly large amounts of hemagglutinating antibodies to three types of enteropathogenic E. ¢oli and also neutralizing antibodies to type 1 poliovirus. Antibody levels were measured in milk, infant and maternal sera, and supernatants from 20 per cent suspensions of stool. Newborn stool antibody titers were usually two- to eightfold lower than those in milk, averaging fourfold lower for antibacterial and only twofold lower for antiviral activity (in the one infant tested at 5 weeks, there was still only a twofold difference): Stool antibody levels correlated well with those of milk, but poorly with those of infant and maternal sera. Coproantibodies were not found in bottle-fed infants, except for traces of antiviral activity in meconium when infant serum antibody titers were high. Antibodies in milk and stool were specific and relatively resistant to trypsin. Bacterial hemagglutinins were sensitive to mercaptoethanol, whereas the viral neutralizing activity was not. Stool extracts were concentrated tenfold for immunodiffusion studies. Transitional and later stools from breast-fed infants were found to contain gamma-l-globulins similar to those in milk, including IgA and small amounts of IgM. No h u m a n immune globulins were detected in stools from artificially fed infants.
61. The etiology of acquired hemolytic anemia
Abstracts
94 3
Wolf W. Zuelzer, Cyril S. Stulberg, Robert H. Page,* Jos~ Teruva,* and A. Joseph Brough,* Child Research Center of Michigan, Children's Hospital of Michigan, and Wayne State University School of Medicine, Detroit, Mich. Clinical, pathologic, and virologic observations in children with acquired hemolytic anemia (AHA) will be presented, indicating the consistent association of hemolysis, irrespective of the presence of autoimmune phenomena, with active disseminated viral infection attributable to the agent of eytomegalic inclusion disease ( C I D ) . The extent of viral activity in the R.E. system became apparent when a morphologic study of the evolution of the classical cellular lesion disclosed recognizable early and intermediate stages of infection at the cellular level and thus led to the demonstration of vast numbers of virus-infected cells in lymph nodes and other tissues at time of active hemolysis. Acute transient AHA was observed chiefly in association with primary CID in otherwise normal infants showing the classical syndrome of generalized infection, hepatitis, or a theretofore unrecognized milder form manifested by generalized lymphadenopathy, hemolysis, and at times, a characteristic rash. The virus as an agent with latency and low virulence meets the theoretical requirements for an etiologic agent sustaining the chronic, intermittent course of AHA. In a high percentage of subjects with chronic intermittent AHA, evidence of impairment of immune mechanism was found, which seemed to account for the persistent or recurrent viral activity, on the basis of a disturbed hostvirus equilibrium. These observations led to the conclusion, supported by serologic-hematologic findings, that autoantibodies in AlIA represent a variable secondary response to viral infection in the R.E. system.
METABOLISM
62. Temporary induction by ethanol of normal glycogenolysis in patients with liver glycogen disease Charles U. Lowe, Luis L. Mosovich,* and Valiollah Abbassi,* State University of New York at Buffalo, Buffalo, N. Y. Observations made during alcohol infusion in 7 patients with hyperlacticemia and hypoglycemia due to liver glycogen disease ( L G D ) challenge the causal relation between enzyme defects observed in vitro and metabolic abnormalities in the whole patient. Oxidation of E t O l i to acetaldehyde is catalyzed by alcohol dehydrogenase with reduction of D P N to D P N H . In a coupled reaction, pyruvate is converted to lactate with regeneration of DPN. When alcohol is consumed
9 44
Society for Pediatric Research
after a fast to remove liver glycogen, lactate and FFA levels in blood rise; the D P N H produced inhibits the conversion of UDP-gal to UDP-glu and glutamate to alpha ketoglutarate. Galactose removal from blood is markedly delayed and gluconeogenesis from amino acids is inhibited. I n contrast with predictions, when acohol was infused into these 7 patients, no hypoglycemia developed; lactate and pyruvate levels fell to normal; FFA failed to rise; alcohol levels ranged from 0 to 22 rag. per cent (controls 90-120 mg. per cent) and there was no interference with galactose removal. In addition, their abnormal hyperglycemic response to glucagon was now entirely normal. These data suggest that alternate pathways for glyeogenolysis as well as alcohol oxidation should be sought; in addition, they challenge conventional thinking in relation to LGD, raising questions as to the significance of observed abnormalities in glucose-6-phosphatase activity and recommend the use of caution in assigning a causal role to abnormalities in enzyme function demonstrated by in vitro studies.
63. Regulation of pyruvate metabolism during development Everett Lovrien,~ Gerald Reaven, ~" and Robert E. Greenberg, Stanford University School of Medicine, Palo Alto, Calif. The importance of various mechanisms controlling formation of glucose from pyruvate is not well defined. As a means of delineating the role of some factors controlling gluconeogenesis during development, the effects of age, fasting, and administration of hydrocortisone on the metabolism of pyruvate by slices of kidney and liver were investigated. Two approaches were used: (1) incorporation of C 14 from pyruvate2-C 1~ into glucose, lipid, glycogen, and CO2, and (2) measurement of activities of pyruvate carboxylase (pyr-C) and phosphoenolpyruvate carboxykinase (PEP-C), which catalyze the first step in gluconeogenesis, conversion of pyruvate to phosphoenolpyruvate. Conversion of pyruvate-C 14 to glucose-C 14 was minimal in slices of fetal kidney and liver, whereas incorporation of (314 into lipid was greater in both tissues during fetal life. Activity of pyr-C and PEP-C was low in both tissues during fetal life, increasing with age. Gluconeogenesis increased in liver slices from newborn and weakling rats following a fast and hydrocortisone; activity of PEP-C similarly increased, while that of pyr-C did not change. Ilydrocortisone increased pyruvate conversion to glucose in kidney slices only from weanling rats, while fasting had no effect at any age. Activity of pyr-C increased with hydrocortisone and fasting only in kidney slices from newborn and weanling rats, respectively, while activity of PEP-C increased following hydrocortisone and fasting in kidney from both age groups. These studies reflect the complexity of mechanisms c o n -
November 1965
trolling gluconeogenesis and indicate that conversion of pyruvate to phosphoenolpyruvate is not the only rate-limiting step in gluconeogenesis during development.
64. Renal phosphate clearance and bicarbonate excretion in the ]etal lamb Fred G. Smith, Jr., Birger O. Tinglof,* and Forrest H. Adams, University of California School of Medicine, Los Angeles, Calif. Neonatal tetany with associated hyperphosphatemia may be related to transitory hypoparathyroidism, renal insensitivity to parathormone, or both. Twelve lambs near term were delivered by cesarean section with special precaution to maintain an intact placental circulation. The fetalmaternal preparation was monitored by measuring arterial pressures, blood pH, pCO2, and body temperatures. Fetal inulin, phosphate, and calcium clearances were measured before and after the intravenous administration of parathyroid extract (0.1 unit per minute per kilogram). In all experiments the clearance of phosphate was calculated as the ratio of the observed clearance to that of inulin. In all fetuses the control phosphate clearances were extremely low (0 to 0.02 c.c. per minute per kilogram) in spite of high serum phosphorus concentrations (mean 9.9 mg. per 100 c.c.). Following parathyroid extract the phosphate clearances ( C p / C I , x 100) changed significantly (0.03 to 0.09 c.c. per minute per kilogram, P < 0.01) demonstrating that the fetal kidney does respond to parathyroid extract. Bicarbonate excretion rates were also studied in 5 fetuses before and after acetazolamide. Bicarbonate excretion increased rapidly from a mean of 1.8 #Eq to 16.1 ~Eq per minute following enzyme inhibition suggesting an active carbonic anhydrase mechanism in the fetal kidney. These studies demonstrate that fetal hyperphosphatemia is due to factors other than insensitivity of the renal tubule to parathormone.
65. Familial heterotopic calcification and byperpbospbatemia unresponsive to parathyroid extract Warren F. Dodge, ¢ Luther B. Travis, ~ and Manuchehr Assemi,* University of Texas Medical Branch, Galveston, Texas Introduced by C. William Daeschner T h e reported cases of heterotopic calcification, accompanied by hyperphosphatemia, normocalcemia, and absence of renal insufficiency, are few.