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This communication introduces a special lyophilization process for selenium determination by fiuorometric methods. It permits a small sample volume, with several modifications including a single test

The effects of hypophysectomy and subsequent administration of growth hormone and/or L-thyroxine on thermotropic properties of State 3 respiration (ADP-induced), cholesterol, phospholipid and fatty acid composition of phospholipid fraction were examined in myocardial mitochondria of rats. Temperature-dependence of 1-anilino-8-naphthalene sulfonate fluorescence was determined in vesicles prepared from lipids of heart mitochondria. Transition temperature obtained from the Arrhenius plots of respiration occurred at 21 and 24 degrees C for heart mitochondria of normal and hypophysectomized rats, respectively. Most notably, after hypophysectomy the rate of respiration was lower below 24 degrees C, but was progressively higher above that temperature when compared to normal rats. The energy of activation was 148 and 36% larger below and above the transition temperature, respectively. Growth hormone restored almost completely the energy of activation and respiratory rates to normal levels. Administration of L-thyroxine, with or without growth hormone, did not significantly change the rate of respiration but decreased the transition temperature to 17.7-17.9 degrees C. Lipid and phospholipid content, as well as percent distribution of phospholipids and their fatty acid composition were not statistically different among the different groups of rats. Only cholesterol content was increased after hypophysectomy. Administration of growth hormone and thyroxine did not significantly change the total unsaturation index of fatty acids, but growth hormone increased the content of arachidonic acid (20 : 4) by 70% but decreased the docosahexaenoic acid (22 : 6) three times which may have a beneficial effect on mitochondrial membranes. These and other results suggest that hormones exert different effects on subcellular organelles in different tissues, like heart and liver.

The effects of hypophysectomy and subsequent administration of bovine growth hormone (0.1 IU/100 g body wt) and l -thyroxine (5 μg/100 g body wt) on respiration, energization-dependent fluorescence of 1-anilino-8-naphthalene sulfonate, NADH dehydrogenase, energy-independent nicotinamide nucleotide transhydrogenase, and succinate dehydrogenase activities were investigated in submitochondrial particles of rat liver. Hormones were injected daily for 7 days. Hypophysectomy decreased the respiratory rate with NADH or succinate and the activities of the three enzymes. Administration of growth hormone increased the respiration but showed selectivity toward NADH. Thyroxine increased the respiration more than growth hormone did with both substrates. Growth hormone increased the activities of NADH dehydrogenase and transhydrogenase whereas thyroxine increased the activity of only succinate dehydrogenase. After growth hormone treatment transhydrogenase activity was increased to about three times that of controls which may have significance in some processes mediated either directly or permissively by growth hormone. When both hormones were injected together, there was a significant decrease in the thyroxine-dependent rise in respiration on succinate as well as the growth hormone-dependent rise in enzyme activities. Fluorescence yield of 1-anilino-8-naphthalene sulfonate in unenergized submitochondrial particles remained unchanged independent of the hormonal status. Energization with succinate or NADH increased the fluorescence yield by about 2–20 times. Several parameters of energizationdependent fluorescence were decreased after hypophysectomy. In restoring these parameters, growth hormone and thyroxine showed specificity toward the energization substrate NADH and succinate, respectively. From the present results we conclude that (a) growth hormone and thyroxine regulate mitochondrial activity by affecting different segments of the respiratory chain, namely Complex I and Complex II, respectively, and (b) growth hormone and thyroxine exert moderating effects on one another.

The effects of hypophysectomy on respiration and ATPase activity of mitochondria of rat heart and liver were investigated. Hypophysectomy decreased significantly the rate of State 3 respiration and ATPase activity of liver but not of heart mitochondria. Heart cytochrome oxidase activity was also not affected. ADP:O and respiratory control ratios were not affected. Administration of bovine growth hormone (bGH) to hypophysectomized rats partially restored the rate of respiration and ATPase activity of liver mitochondria. However, hypophysectomy decreased heart mitochondrial protein synthesis as measured by in vivo and in vitro incorporation of radioactive leucine, but did not affect the in vitro uptake of the amino acid. Administration of bGH restored the incorporation capacity to the normal level. Similar effects on liver mitochondrial protein synthesis have already been reported. These data suggest that the effects of growth hormone on mitochondrial function may vary with the two tissues.

γ-Glutamyl transpeptidase (GGTP) activity in normal amniotic fluids and corresponding maternal sera obtained at various gestational periods was measured. The ontogenic pattern of enzyme activity in amniotic fluid is very similar to alpha fetoprotein (AFP). However, the levels of these two proteins behaved differently in corresponding maternal sera. Also, in amniotic fluids obtained from pregnancies with neural tube defects (NTD), only AFP concentration was abnormally high whereas GGTP activity was normal.

Thermotropic effects on the kinetics of glucose-6-phosphatase (D-glucose-6-phosphate phosphohydrolase, EC 3.1.3.9) activity of hepatic microsomes from normal and alloxan-diabetic rat liver were investigated by determining V, Km and Ki (substrate inhibition) values. Influence of deoxycholate (0.1%) and 1-anilino-8-naphthalene sulfonate (2.5 mM) on the kinetics was also evaluated. 1. Substrate inhibition occurred at 0.06 M for the enzyme from normal rats and at 0.0-0.025 M for the enzyme from diabetic rats. 2. The enzyme from diabetic rats showed a transition that extended between 22.7 and 27 degrees C in the Arrhenius plot (log V vs. T-1) instead of at 19.5 degrees C. 3. Deoxycholate increased the V value of both enzymes without affecting substrate inhibition at all the temperatures but did not completely abolish the transition in the Arrhenius plot of the enzyme from diabetic rats. 4. 1-Anilino-8-naphthalene sulfonate eliminated substrate inhibition and activated the enzyme of normal rats above 27.5 degrees C by increasing both V and Km values. Below this temperature, the enzyme showed biphasic or allosteric kinetics. At low substrate concentrations it was activated as both V and Km values were increased. The enzyme from diabetic rats, on the other hand, was activated at all the temperatures and exhibited linear kinetics. 5. Binding of 1-anilino-8-naphthalene sulfonate to the microsomal fraction increased with decreasing temperature as revealed by the increase of relative fluorescence. The microsomal fraction of diabetic rats showed a more anomalous fluorescence response between 13-18 degrees C. 6. Enthalpy changes for glucose 6-phosphate binding to the inhibition site were slightly larger than binding to the active site. Calculated entropies of activation for transition state complex of glucose-6-phosphatase reaction were fairly large and negative. The free energy of activation (28-30 kcal/mol) was independent of temperature and experimental conditions. 7. In the microsomal fraction (total as well as rough), phospholipid content and fatty acid unsaturation index of phospholipids were decreased after diabetes. The level of free cholesterol remained unchanged but the molar ratio of cholesterol to phospholipid increased. The different thermal response and 1-anilino-8-naphthalene sulfonate interaction to the enzyme from diabetic rat and liver could be ascribed to the altered lipid environment of the enzyme on the endoplasmic reticulum membrane.

Salt-washed and unwashed and aluminum oxide treated microsomes of human liver, and washed microsomes of liver of normally fed, 1 day fasted and alloxan diabetic rats were subjected to polyacrylamide gel electrophoresis in presence of sodium dodecylsulfate and urea. Unwashed and aluminum oxide treated microsomes showed several protein bands, but salt-washed microsomes gave only one major protein band. This major protein band which appeared in all the preparations showed glucose-6-P activity on incubation of the electrophoresed gel with the substrate and subsequent staining with ammonium sulfide. Molecular weight of this protein was estimated to be 63,000 ± 6,800. Glucose-6-P and pyrophosphatase activities were measured after subjecting lyophilized microsomes, aluminum oxide treated supernatant and residue fractions of human and rat liver to ionizing radiation in vacuo at different doses (Mrads). There was no significant difference between either the surviving fraction of glucose-6-P and pyrophosphatase activities or among the different preparations of human and rat liver at a given radiation dose. Computer analysis of the pooled data gave a value of 9.16 Mrads for 37% survival of the activity which corresponds to a molecular weight of 70,000.

A newborn infant had an unusual reversible complication caused by light therapy for hyperbilirubinemia. A dark grey-brown color of the skin, urine, and serum developed on the fifth day, after 24 hours of light therapy. Serum bilirubin (direct and total), SGOT, and LDH were elevated. The infant recovered, and bilirubin and enzymes returned to normal after a period of three weeks. A possible pathologic mechanism for this complication is suggested.

The emission maximum of human growth hormone, excited at 280 nm, in aqueous solution (pH 7.5) and in the presence of 6 M guanidine is at 348 nm. Rates of increase of tryptophanyl fluorescence of human growth hormone with ethanol or dioxane (up to 30%, by vol.) are identical to those shown by N- acetyl- l - tryptophanamide . Perturbation of tyrosyl fluorescence by ethanol is identical to that of N- acetyl- l - tyrosinamide whereas by dioxane it is only 39%. Tryptophanyl fluorescence of human growth hormone is quenched by about 35% between pH 9 and II in the aqueous solution but not in the presence of 6 M guanidine. These results suggest that the single tryptophanyl residue of human growth hormone is accessible to solvent molecules and may be situated close to tyrosyl residue(s).

Summary A patient with galactosemia presenting with bilateral pedal gangrene and ascites is presented. The cause of gangrene is obscure, but it may be presumed to be due to sepsis associated with multiple small venous and capillary thromboses.

l-[U-14C]Leucine incorporation into the trichloroacetic acid-precipitable fraction of mitochondria after incubation with succinate as energy source was significantly higher with normal rat liver mitochondria than with hypophysectomized rat liver mitochondria. The incorporation was sensitive to inhibition by chloramphenicol, terramycin, dinitrophenol, and NaCN, but not to RNase and cycloheximide. The differences in the rate of incorporation between the two types of mitochondria were not abolished when the endogenous energy source from succinate was replaced by an external ATP-generating mixture or when cold trichloroacetic acid precipitate was extracted with hot 5% trichloroacetic acid or when high concentrations of unlabeled leucine (up to 200 mm) were added. Uptake of l-[U-14C]leucine, in the absence of incorporation, was higher with hypophysectomized rat liver mitochondria than with normal rat liver mitochondria. However, when mitochondria were washed extensively after incubation there was no significant difference in the uptake. Growth hormone treatment of hypophysectomized rats for 5 days significantly increased both in vivo and in vitro incorporation of leucine. The in vitro stimulating effect of growth hormone became apparent as early as 15 min and was significant 30 min after a single intravenous injection of the hormone. These results suggest that growth hormone has a significant effect on liver mitochondrial protein synthesis.

Uptake of 3 H-acetyl human growth hormone ( 3 H-HGH) and 3 H-acetyl bovine growth hormone ( 3 H-BGH) into slices of hypophysectomized rat liver and human liver (obtained from autopsy) has been measured both as a function of time and concentration of the labeled hormones in the incubation medium. Uptake kinetics as a function of concentration of 3 H-HGH or 3 H-BGH in rat liver slices followed a sigmoid curve. Similar sigmoid curve was obtained when increasing concentrations of 3 H-HGH were incubated with human liver slices. In contrast, uptake kinetics of 3 H-BGH in human liver slices followed a hyperbolic curve. Addition of unlabeled HGH or BGH into the incubation medium reduced the uptake of 3 H-HGH in rat liver slices but in human liver slices only human growth hormone competed. This binding-site difference along with the reported molecular differences between human and bovine growth hormones may well explain the different biological specificities of the two hormones.

3 H-Acetyl human growth hormone ( 3 H-HGH) (∼60 μg) was injected intravenously into hypophysectomized rats. Radioactivity appeared in all the subcellular fractions of liver as early as 2 minutes after the injection. Until 10 minutes postinjection virtually all of the tissue radioactivity was precipitable with trichloroacetic acid, whereas at 20 and 40 minutes after injection all of the radioactivity was soluble in trichloroacetic acid. Liver uptake (cpm/mg protein) peaked at about 20 minutes postinjection. The appearance of radioactivity in the crude nuclear fraction closely followed that of the whole homogenate. Radioactivity in the microsomal and mitochlondrial fractions was highest at the early time periods (up to 10 minutes) and then declined progressively. The radioactivity in the final supernatant or cytoplasmic fraction increased slowly, peaked at 20 minutes, and remained the highest. Pretreatment with unlabeled or 14 C-labeled HGH significantly reduced the mitochondrial and microsomal uptake of 3 H-HGH. In contrast with these results, when 3 H-HGH was added in vitro just before homogenization and fractionation of liver, most of the radioactivity (77%) appeared in the cytoplasmic fraction. These results are interpreted to mean that the appearance of 3 H-HGH in the particulate fractions at these early time periods may have functional significance, and that important initial binding sites of growth hormone in the liver cell may be mitochondria and microsomes.

Glucose 6-Phosphatase and inorganic pyrophosphatase were assayed in homogenates prepared from livers obtained at autopsy from 19 children and 18 adults. There was a remarkably good correlation between these two enzyme activities. Both activities were significantly higher in children than adults. Experiments with rats indicated that fasting increased both activities, that actinomycin D administration did not reduce the effect of fasting, that fasting hypophysectomized rats did not increase either activity, and that long-term growth hormone administration reduced both enzyme activities in hypophysectomized rats. These data support the concept that both human and rat liver glucose 6-phosphatase and inorganic pyrophosphatase are two activities of the same enzyme.

Potassium chloride concentrations of 100–150 mM were shown to inhibit (20–40%) human and rat liver microsomal glucose-6-phosphatase when the substrate was reduced to physiological concentration. When the enzyme was solubilized by treatment of microsomes with Al2O3 the inhibition could be observed at greater substrate concentrations. Since potassium deprivation is associated with hyperglycemia in both animals and man, these observations may have physiological significance.

13 short children aged 7-13 years who had a retarded bone age and low hair zinc concentration (under 140 micrograms/g) were treated with oral zinc supplements for a year. There was a significant increase in the growth rate in the children whose hair zinc concentration increased. Growth hormone, testosterone and somatomedin C also increased after oral zinc supplementation. Data from 755 short healthy children who have attended our Growth Clinic are presented which describe their hair and serum zinc concentration at different ages. The data indicate a decline in hair zinc concentration after birth with a gradual increase at age 4-6 years, finally reaching adult normal levels after adolescence.

Effects of hypophysectomy and subsequent growth hormone administration on mitochondrial enzymes of the urea cycle were investigated in rat liver. Hypophysectomy increased the activities of the two mitochondrial enzymes, carbamyl phosphate synthetase and ornithine transcarbamylase but not of the cytosolic enzyme, argininosuccinate synthetase. The activity of mitochondrial phosphate dependent glutaminase was not affected. Administration of bovine growth hormone (100 μg/100 g body weight) for two weeks decreased the activities of carbamyl phosphate synthetase and ornithine transcarbamylase almost to the normal level. These results suggest a specific effect of growth hormone on mitochondrial enzymes of the urea cycle and serve to explain the increased urea formation in hypopituitarism.

Using atomic absorption spectrophotometry, we have assayed zinc and copper concentrations in the hair of children with typical achondroplasia, familial short stature, or constitutional growth delay, and of unaffected parents of achondroplastic children and healthy adults of similar age and sex as the parents (Table). Specimens consisted of the terminal 2 to 3 inches of hair, obtained from the posterior scalp. Duplicate 60 to 80 mg hair specimens were digested in concentrated nitric:perchloric (3:1) acid mixture. The results are the mean of at least two determinations per specimen. The achondroplastic children were all confirmed as typical examples by physical examination and radiographs of skull, spine, long bones, and pelvis. Several of the parents were noted to have white flecks in their fingernails, which has been reported to be associated with zinc deficiency. The data indicate significantly reduced zinc and increased copper values, and reduced zinc:copper ratio, in the hair of parents and children with achondroplasia.

Oxygen consumption by circulating lymphocytes of children with isolated growth hormone deficiency was studied before and 6 mo after the start of growth hormone therapy. A plot of percent change in respiration (oxygen consumed per mg protein or μg DNA) against height gain during the therapy showed a linear association (correlation coefficient 0.667–0.756).

A 14-year-old girl and a 13-year-old boy were found to be growth hormone deficient by insulin-arginine stimulation tests, and were also found to be zinc deficient. When oral zinc replacement was given, they both had a significant increase in growth rate which continued for at least 2 years, and subsequent growth hormone tests were normal.

This rare syndrome is characterized by short stature, sparse, fine scalp hair, a pear-shaped nose, long philtrum, normal intelligence, and cone-shaped epi physes of the phalanges with deformities of the fingers.

Electron micrographs of hepatocytes from hypophysectomized rats when compared with those of normal rats showed increased number or crowding of mitochondria per unit cytoplasmic volume with decreased endoplasmic reticulum, decreased mitochondrial matrix density, and the presence of partitioned mitochondria. After hypophysectomy, the recovery of mitochondrial proteins per gram of wet liver, by centrifugation, was increased. The mean half-life value for the turnover of total mitochondrial proteins was doubled when measured by injection of either l -[4,5- 3 H]leucine or l -[guanidino- 14 C]arginine. These changes were reversed towards normal by subsequent treatment with growth hormone. These and other results suggest a role for GH in the maintenance of a normal population of properly functioning mitochondria in liver.

Premature infants have a lower selenium concentration in serum than full-term infants and children. The selenium concentration goes down quickly in infants treated for respiratory distress syndrome without supplementation. One premature infant with bronchopulmonary dysplasia had persistently low concentrations of selenium. Vitamin E supplements did not affect the serum selenium concentration in healthy premature infants. Supplementation with 3 µg/kg of selenium in parenteral fluids prevented the fall in the concentration seen in other infants not supplemented. Premature infants and especially those treated withoxygen may warrant selenium supplementation to the parenteral nutrition solution. Vitamin E supplements alone are apparently not sufficient to prevent selenium deficiency and potential oxygen toxicity.

Zinc levels were measured in hair and serum of boys with constitutional growth delay and familial short status and in several boys before and after oral administration of methyltestosterone. These results show the following: (1) zinc levels in boys beyond stage 3 of genital development are significantly higher than in stage 1 and 2; (2) there is a linear relationship between zinc levels and serum testosterone concentration (up to 250 ng/dL); and (3) methyltestosterone administration raised the zinc concentration in serum and hair, especially in boys with constitutional growth delay. Therefore, increased endogenous production or exogenous supply of testosterone are associated with increased zinc levels. We speculate that the relative testosterone deficiency and hypogonadotropism seen in constitutional growth delay may result in decreased zinc levels, which in turn could cause a further delay in the appearance of secondary sexual characteristics and greater growth retardation.

The effects of hypophysectomy and subsequent administration of growth hormone, thyroxine, insulin, and testosterone were examined in rat liver for the relationship between the thermotropic effects on State 3 respiration (ADP induced) and fatty acid composition of the phospholipid fraction of intact mitochondria as well as of inner membrane vesicles. The Arrhenius profile for energy-linked (succinate) State 3 respiration of mitochondria from hypophysectomized rats lacked the discontinuity at 23.5 °C seen with mitochondria from normal rats. After injections of the hormones the discontinuity representing the transition temperature from gel to liquid crystalline state of lipids occurred at different temperatures: 18.5 °C for growth hormone, 26.0 °C for thyroxine, 19.5 °C for growth hormone + thyroxine, 27.6 °C for insulin, and 25.3 °C for testosterone. The energy of activation between 37.5 and 23.5 °C was 1.9 times greater for hypophysectomy than for controls. Growth hormone was the most effective in restoring the energy of activation to normal, above as well as below transition temperature. The effect of thyroxine appears to be due to a larger stimulation of the State 4 respiration than that of growth hormone, insulin, or testosterone, especially at higher temperatures. Phospholipids extracted from intact mitochondria or inner membrane vesicles of hypophysectomized rats contained less arachidonic acid (20:4) and more linoleic acid (18:2) than those of normal rats. In addition, the contents of some of the minor fatty acids were also changed. Calculated unsaturation index showed an 18.8 and 14.9% depletion in unsaturation in whole mitochondria and inner membranes, respectively. Among the different hormones used to treat the hypophysectomized rats, growth hormone was the most effective in restoring the transition temperature and fatty acid composition to normal levels and increasing the gain in body weight. Although the other hormones increased total unsaturation index to some extent, some of the individual fatty acids were affected differently. Good correlation exists between the unsaturation index of mitochondrial fatty acids and transition temperature of State 3 respiration. These results strongly suggest a role for the hormones, particularly growth hormone, in the control of mitochondrial membrane fluidity of hypophysectomized rat liver, through fatty acid composition of phospholipids.

Glucose-6-phosphatase activity and associated inorganic pyrophosphatase and pyrophosphate-glucose phosphotransferase activities of human liver microsomes was increased more than two-fold when the pH of the microsomal suspension was adjusted to 9.5 by Al2O3 powder and stirred overnight. Nearly 68% of glucose-6-phosphatase activity did not sediment at 105,000 × g after this procedure. The lipid content, pH optimum, Km, and antigenic characteristics of Al2O3 treated and intact microsomes were similar, but the enzymatic activity of treated fractions was more stable to heat (30°) and the RNA content was reduced by 50%. Electron microscopy indicated that the treated microsomes were essentially free of ribosomes and fragmented. The treatment appears to have dissociated the microsomal membrane by removing ribosomes, thus exposing more enzymatic active sites.

CAT-SCRATCH fever is usually a nonbacterial regional lymphadenitis that occurs one or two weeks after a scratch by a cat. The course is variable and may last for a few weeks to months. The etiology...

Interest in human nutrition has been gaining rapidly in recent years. From the youngest child and oldest man, from the earliest grade school student to the most sophisticated graduate student, from the politician to the scientist, from the pulpit and at the autopsy table, we are all being bombarded with modern suggestions, theories, and recommendations that will improve human nutrition and prolong good health and life. While the food industry continues to advise the public, scientists lament the lack of nutrition education in medical schools. Parenteral Nutrition At the Maimonides Medical Center Symposium on Nutrition of the Fetus, Infant, and Child, 1973 (Brooklyn, NY, May 1973), Gilbert Forbes, MD (University of Rochester) proposed the goddess "Nutrix" for our consideration. Worship of this goddess by scientists, by millions of Americans, and by advertisers has resulted in a religion based upon faith rather than facts. How we eat and what we order

To understand the early molecular events that lead to some of the effets of growth hormone on liver, a dose of 3H-HGH (18 × 106 cpm, 60 μg) was give i.v. to hypophysectomized rats. After the injection, livers were homogenized and fractionated by centrifugation. Specific activity (cpm/mg protein) and total radioactivity expressed as percent of that in the homogenate were estimated (table). 3H-HGH in the homogenate was estimated by precipitation with TCA, and with Na2SO4 as 3H-HGH antibody complex with and without unlabeled hormone. Up tp 10 min most homogenate was antigenically and 3H-HGH in the homogenate was antigenically similar to the injected dose. In contrast, when 3H-HGH was added in vitro before homogenization most radioactivity (77%) appeared in the cytoplasm and the rest distributed almost eqully among the other fractions. These results indicate (1) the early rapid uptake by the particulate fractions may be of functional significance and the important initial binding sites may be mitochondria and microsomes; (2) the movement of 3H-GH in liver cell may be from the microsomal and mitochondrial fractions into cytoplasm; (3) after 10 min 3H-HGH is degraded or deacetylated.

17 children between 5 and 15 years of age received mazindol for one year (2 mg daily). Their height was carefully evaluated by 2 physicians at 6 month intervals, and they were observed for the following year without any therapy. In each case, the growth rate slowed during mazindol administration (0.6 in/yr) compared to the previous year (1.5 in/yr). Assays of growth hormone during mazindol administration demonstrated reduced responses to insulin hypoglycemia and arginine (2 patients) and exercise (3 patients). It seems likely that mazindol reduces growth by increasing nor-epinephrine in the ventromedial nucleus of the hypothalamus, and it is interesting that increasing dopamine in that nucleus stimulates growth hormone release.

Perinatal stress may induce a surge of hormonal responses which includes the thyroid hormones. Information is inadequate with regard to the surge of thyroid hormones at the time of delivery as it relates to the type of CS. We studied 33 motherinfant pairs who had elective (E) or primary (P) CS. The infants studied had (x) gestational age of 38.0 wk. All thyroid hormone measurements made on venous cord blood obtained immediately after delivery on two CS groups follow (X±SEM): The following is comparative serum TSH concentrations (X±SEM) of subgroups of infants who underwent CS delivery with and without preceeding labor: The data suggest that labor may suppress TSH and the events surrounding the primary CS stimulates this surge at the time of delivery. The mechanism operative is not clear and needs further study.

A pituitary evaluation was carried out in a 12-year-old girl who had early puberty, short stature, optic nerve hypoplasia, and agenesis of the corpus callosum. Her growth hormone (GH) response to insulin-induced hypoglycemia and arginine infusion was blunted. Basal and stimulated levels of prolactin were elevated. The association of GH deficiency and hyperprolactinemia suggests a disruption of the dopaminergic modulation of these hormones. The facial features seen in our patient, such as hypertelorism, V-shaped frontal hairline, and cleft nose and upper lip, are enough to justify the diagnosis of median cleft face (MCF) syndrome. However, the optic nerve hypoplasia and the GH deficiency are characteristics of septo-optic dysplasia, and, to our knowledge, they have never been described in patients with MCF syndrome. Our case fulfills the diagnostic criteria of both, representing a link between both ends of this spectrum.

Zn deficiency has been reported in infants with BD and in children born with achondroplasia. 304 families were studied comparing parental age, Zn nutritional status and infant weight at the time of birth. There is a statistically significant correlation (p < .01) between infant hair Zn and maternal age at time of birth. The decrease in infant hair Zn seen with teenage (14-20) and elder mothers (31-40) may be related to the incidence of increased BD which occurs in these groups. Infant hair Zn was lowest in the heaviest infants and in a group with BD made up of porencephaly, anencephaly and microcephaly as the presenting sign (N=4; Zn=134±10). The heavier children may represent the infants of potential diabetic mothers (who have more BD). Placental Zn was 18-19 ug/g and did not correlate well with maternal or infant hair Zn. Finally, diet did not explain parental Zn status since paired deficiencies did not occur. Therefore, an association between Zn levels, high-risk age groups and those children with BD is apparent.

Serum was collected from hypophysectomized rat (HyR) 30 min. after an I.V. injection of hGH. The serum was used to prepare an antiserum (AS) in a rabbit. AS was adsorbed with lyophilized HyR serum (5 mg/ml) and centrifuged to obtain adsorbed antiserum (AAS). Immunoelectrophoresis (pH 9.5) using AS produced a number of lines with normal, HyR serum and Hy + hormone-treated serum but no line with hGH or insulin. AAS did not react with HyR serum but produced a single identical line (at beta globulin) with normal or Hy + GH treated serum. This precipitin line which was also present when whole AS was used may correspond to somatomedin produced in response to hGH. A similar and identical precipitin line was obtained with HyR + bGH treated serum but not with normal human serum. These results suggest that (1) hGH and bGH produce immunologically similar somatomedin(s) in the rat, (2) human somatomedin is immunologically different from rat somatomedin. This laboratory has already shown differences in the kinetics of bGH and hGH uptake by human liver slices.

Growth hormone deficiency has been reported in 10-20% of Russell-Silver dwarfs. Two siblings with all the classical features of Russell-Silver Syndrome were low in growth hormone (insulinarginine stimulation test), and blood and hair zinc (Zn). They received 50 mg elemental Zn orally each day for 2 months and 50 mg weekly for 4 more months. Growth hormone and Zn determinations were repeated at 6 months: It appears there is a direct relationship between the growth hormone response and Zn nutritional status. We have data indicating that growth hormone therapy increases hair Zn and decreases urinary Zn in children with growth hormone deficiency. These two children provide new data indicating that there are patients whose growth hormone production is affected by their Zn nutritional status.

Concentration of zinc in hair and serum of children with short stature (familial or constitutional) was measured by atomic absorption spectrophotometry. Some of them were treated with methyltestosterone (MT), 10 mg. daily. Our Zn values were similar to those reported for normal children. Nevertheless, children receiving MT had higher Zn levels than the children without medication. Since Zn deficiency is associated with growth retardation, these results raise the question of whether increased Zn retention may be one of the mechanisms by which androgens accelerate growth. We were unable to find any significant difference between the Zn levels from children with familial short stature and those from children with constitutional growth delay.

Concentration of zinc (Zn) in hair, urine and serum were measured by atomic absorption spectrophotometry in one 14-month old girl with typical features of Fetal Alcohol Syndrome (FAS) (peculiar facies, congenital malformations, pre and post natal growth deficiency). She was born to an alcoholic woman who continued drinking heavily throughout her pregnancy. Zinc level was low in hair (66 μg/g) and urine (266.8 μg/g of creatinine) but normal in serum(95 μg%), reflecting probable chronic depletion of Zn. (Normal values are: hair 193 ± 18 μg/g, serum 75-160 μg%, urine 400-600 μg/g.) It has been shown that the offspring of Zn-deficient rats have marked growth retardation and high incidence of congenital malformation, some of which are similar to those seen in FAS. Alcoholic patients have been found to lose increased amounts of zinc in urine. Therefore, it is tempting to speculate that congenital Zn deficiency plays an important role in the pathogenesis of this syndrome.

Since there is a relationship between prostaglandin metabolism, Se and glutathione peroxidase (GSH-PX), and diabetics are reported to have low levels of Se and prostacyclin, we have wondered whether some of the problems which occur in IDM (cardiomyopathy and persistent fetal circulation) might be related to Se deficiency. We studied Se in whole blood of IDM (within 1-2 days after birth) and their mothers. The IDM had a birth weight (mean±SD) of 3857±1075 gm, gestational age of 37.8±1.5 wks, 1 and 5 minute Apgar scores of 7.3±2.7 and 8.8±1.7 respectively. Correlation between the maternal and infants' blood Se concentration in IDM group was not significant. Comparison between IDM, normal full term newborn (NB) infants, and the mother of IDM follows: This preliminary study did not find unusually low Se in whole blood of IDM or their mothers, but because of the potential importance of Se in IDM, a larger study is still indicated.

We have shown in laboratory animals that GH stimulates several mitochondrial processes like protein synthesis, bulk-protein turnover, levels of eytochromes and enzymes, and respiratory activity. We have presently studied respiratory activity (RA) in white cells (WC) and thymidine uptake in LY of children before and after GH therapy. RA (ngm of O2/mg protein or/μg DNA) was measured in WC. prepared by layering whole blood on metrizoate-dextran medium, with a Clark-Oxygen electrode in Krebs-Ringer solution containing glucose. There is significant correlation (r =0.8) between percent increase in respiration due to GH therapy for 1 month and height Rain after 4-8 months of therapy in 16 children. This correlation may have significance in the early prediction of growth-promoting effect during therapy. LY, isolated by layering whole blood over Lymphoprep, showed an increase in WBC, LY count and rosette forming LY in 11-12 children but showed a decrease in 5-6 children. There is a significant decrease in thymidine uptake by unstimulated LY in all 12 children. Therefore, stimulation index (uptake with stimulation by phytohemagglutinim/uptake without stimulation) increased. These results suggest that GH therapy may reduce basal rate of DNA synthesis in LY.

The oral administration of Clonidine, (C) a selective Alpha-Adrenergic agonist has been shown to increase growth hormone(GH) secretion in children and has been proposed as one of the reliable tests of GH reserve. The effects of chronic administra tion of C on growth have not yet been reported. We have now evaluated the effects of C administration on the rate of growth (RG) and somatomedin C (SM-C) levels in normal prepubertal children with short stature. Ten children (4-9y) with normal RG and normal GH reserve as assessed by the C test and SM-C determination were the subjects of this study. C was administer ed orally to these children as a single dose (0.15mg/m2/body surface area) every night during a 6-month period. Height, weight and blood pressure were measured. Plasma SM-C was determined before and one week after the last C dose. No side effects were observed during the 6-month period. The administration of C during that period produced a significant increase in the RG and SM-C levels. We conclude that C-administration could be used as an alternative treatment modality for normal children with short stature

• A mildly obese 15-year-old boy had short stature with rounded facies and short, stubby hands and toes. He had the fully expressed syndrome of pseudohypoparathyroidism but was the only member of his family who had all the somatic characteristics of this disease. The serum parathyroid hormone level was substantially elevated. Urinary excretion of cyclic adenosine monophosphate and phosphate failed to increase following intravenous infusion of parathyroid hormone. However, he did not have hypocalcemia. The present entity is probably a transient form of pseudohypoparathyroidism with partial responsiveness of skeletal adenyl cyclase to parathyroid hormone. ( Am J Dis Child 129:1092-1095, 1975)

504 EARLY PUBERTY IN KLINEFELTER SYNDROME DUE TO A CHORIONIC GONADOTROPIN (hCG)-PRODUCING TUMO