Particulate concentrations in pulmonary alveolar proteinosis. Quantitative birefringent particle counts per 10 high-power fields in lung tissue were correlated with age, sex, and occupational exposure in 37 cases of pulmonary alveolar proteinosis. Counts were significantly higher in pulmonary alveolar proteinosis cases in both regions of alveolar proteinosis (47 +/- 11) and perivascular and peribronchiolar areas (dust retention areas) (275 +/- 65) than in 13 controls (5 +/- 3 and 79 +/- 23, respectively). Of a total of 8619 particles, 4817 were less than 1 micron, and 3771 were 1 to 10 microns in diameter. Fifty-nine percent were round, 19% fibrous, and 22% irregular. When analyzed individually, 20 of 37 cases (78%) had alveolar particle counts significantly higher than controls, and 10 of 26 cases had dust-retention area counts greater than controls (P less than 0.050). Known or possible occupational exposure was ascertained in 13 cases. In the remaining 24 cases insufficient occupational information was available. The mean age of the pulmonary alveolar proteinosis patients was 33 +/- 4 years, and of the controls, 40 +/- 7 years, and there was a male-to-female ratio of approximately 3:1. We propose that many cases of pulmonary alveolar proteinosis will ultimately be shown to be associated with environmental exposures to fine dusts or fumes.


Selenium levels in human blood, urine, and hair in response to exposure via drinking water. Blood, hair, urine, and tap water samples were obtained from participants in a population exposed to varying amounts of selenium via water from home wells. Concentrations of selenium in urine and hair produced significant positive correlations with well-water selenium levels. Blood selenium with well-water selenium did not produce a significant correlation. Urine selenium correlation with hair selenium and urine selenium correlation with blood selenium were found to be significant. No significant correlation for hair selenium with blood selenium was found. We conclude that blood selenium alone is not an adequate monitor of selenium exposure or body burden at levels below chronic toxicity.


Modification of methylmercury neurotoxicity by vitamin E. The protective effect of vitamin E on the neurotoxicity of methylmercury was investigated by means of light and electron microscopy. Young male golden hamsters were exposed to 2.0 ppm methylmercury chloride with or without concurrent administration of vitamin E (2.0 ppm). No toxic symptoms were observed in the vitamin E-protected animals while all the animals given methylmercury alone developed severe symptoms of methylmercury poisoning. Light microscopy revealed no significant neural damage by mercury in those animals exposed to methylmercury/vitamin E while significant neuronal necrosis could be demonstrated in both the cerebellum and calcarine cortex of the methylmercury-treated animals. Besides some accumulation of lysosomes, electron microscopy also demonstrated remarkable intactness of the cellular organelles in the nerve cells without neuronal necrosis. It appears that vitamin E has a strong protective potential against the toxicity of methylmercury.


Asbestos cement dust inhalation by hamsters. Two groups of 96 male Syrian golden hamsters were exposed to respirable asbestos cement aerosol at concentrations of approximately 1 and approximately 10 micrograms/liter, respectively, 3 hours/day, 5 days/week. Average fiber counts ranged from 5 to about 120 fibers/cm3. Each group was randomly divided into six subgroups of 16 animals. The first subgroup was sacrificed after 3 months of exposure, the second after 6 months, and the third after 15 months. The fourth subgroup was withdrawn from exposure after 3 months, observed for an additional 3 months, and then sacrificed. The fifth and sixth subgroups were withdrawn after 3 and 6 months of exposure, respectively, and maintained for observation up to the 15-month exposure point of the third subgroup at which time all surviving animals were sacrificed. All other experimental procedures were similar to those delineated in a previous publication describing the development of an animal model, techniques, and an exposure system for asbestos cement dust inhalation (A. P. Wehner, G. E. Dagle, and W. C. Cannon, 1978, Environ. Res. 16, 393-407). The asbestos cement exposures had no significant effect on body weight and mortality of the animals. Higher aerosol concentration and longer exposure times increased the number of macrophages and ferruginous bodies found in the lungs of the exposed animals. Recovery periods had no effect on the incidence of macrophages and ferruginous bodies. The incidence of very slight to slight fibrosis in the animals sacrificed after 15 months of exposure shows a significant (P less than 0.01) trend when the untreated control group and the 1 and 10 microgram/liter dose level groups are compared, indicating a dose-response relationship. Development of minimal fibrosis continued in animals withdrawn from exposure. No primary carcinomas of the lung and respiratory tract and no mesotheliomas were found.


Lead in red blood cells and in plasma of pregnant women and their offspring. Lead levels were measured in red blood cells and plasma of 75 pregnant women and their infants at delivery. No significant differences were found by comparing maternal and fetal values and a statistically significant positive correlation was evidenced, higher for plasma levels. The results suggest that the diffusible form of maternal lead in plasma influences the lead transferred to the fetus more directly. The highly significant correlation found between the maternal and fetal ratio of lead in plasma to lead in erythrocytes seems to be in favor of inborn factors influencing the equilibrium between the two compartments in which lead is distributed.


Geophagic lead nephropathy: case report. An adult presenting with anemia and seizures was found to have lead poisoning. Chelation therapy undertaken before the source of exposure was known was accompanied by clinical improvement. Recurrence of an excessive body lead burden despite chelation led to the discovery of pica for lead-contaminated garden soil. Lead nephropathy progressed when the geophagia was resumed.


Effects of chronic ingestion of south Louisiana crude oil on mallard ducklings. South Louisiana crude oil was fed to duckling mallards (Anas platyrhynchos) in concentrations of 0.025, 0.25, 2.5, and 5.0% of the diet from hatching to 8 weeks of age to assess the effects of chronic oil ingestion during early development. Growth was depressed in birds receiving a diet containing 5% oil but there was no oil-related mortality. Diets containing 0.25, 2.5, and 5.0% oil impaired avoidance behavior of 6-day-old mallard ducklings when compared with controls or ducklings fed 0.025% oil, but had no effect on open-field behavior of 7-day-old ducklings. Liver hypertrophy and splenic atrophy were gross evidence of the pathological effects of oil in birds on the 2.5 and 5.0% oil diets. Biochemical lesions that occurred included elevation of plasma alanine aminotransferase and ornithine carbamyl transferase activity. Hepatocyte hypertrophy and bile duct proliferation in the liver were noted in birds fed the 2.5 and 5.0% oil diets and tubular inflammation and degeneration in the kidney were noted in birds fed the 5.0% oil diet.


Effects of methylmercury on mitotic mouse glioma cells. The effects of methylmercury on cultured mammalian cells were studied by light and electron microscopy. At 5 x 10(-6) M concentration methylmercury depressed the proliferation of mouse glioma cells almost completely and increased the mitotic indices. In the electron micrographs of the cells treated with 5 x 10(-6) M methylmercury for 4 hr, microtubules as mitotic spindle fiber were absent and chromosomes were scattered in a disorderly fashion in the cytoplasm. From these observations we concluded that exposure to methylmercury inhibits the cell mitosis by blocking the polymerization of tubulin to microtubules and results in the accumulation of the cells on the way of mitosis. Changes in behaviors of other mitotic apparatus and of fibrillar structures other than microtubules are also described.


Sodium and potassium fluxes across the dialyzed giant axon of Myxicola. Resting and stimulated fluxes of sodium and potassium across the giant axon of the marine annelid, Myxicola infundibulum, have been characterized using the technique of internal dialysis. In most respects the ion movements were found to be similar to those in squid axons. Sodium efflux and potassium influx were found to be active, cardiac glycoside-sensitive fluxes, with a variable coupling ratio. However, when [ATP]i was lowered to less than 20 microM by treatment with cyanide and continuous dialysis, or to less than 2 microM by dialysis with glucose following injection of hexokinase, Na efflux and K influx were unaltered. The maintained fluxes were not accounted for by an increased passive permeability of the axolemma, although 30-60% of the Na efflux appeared to be due to Na-Na exchange. An altered form of Na pump operation at low [ATP]i is a more likely explanation than an alternate energy source, or an ATP source proximate to the axolemma. The transient response of 22Na efflux to a change in [22Na]i was found to be much slower than in squid, tau = 360 sec. The efflux delay could only be accounted for by an extra-axonal diffusion barrier, which is probably the basement membrane surrounding the ventral nerve cord.


Ion transport and displacement currents with membrane-bound carriers: the theory for voltage-clamp currents, charge-pulse transients and admittance for symmetrical systems. The standard carrier model for ion transport by a one-to-one mechanism is developed to predict the time-dependent currents for systems that are symmetrical at zero applied potential. The complete solution for ions and carriers bearing any charge is derived by assuming that the concentration of ions in the membrane is low and either that the applied potential is small or that the applied potential affects equally all of the association and dissociation reactions between the ions and the carriers. The response to an abruptly applied potential is then given by the sum of a constant and two declining exponential terms. The time constants of these relaxations are described by the equations derived for neutral carriers by Stark, Ketterer, Benz and Lüger in 1971 (Biophys. J. 11:981). The sum of the amplitudes of the exponentials for small applied potentials obeys a relation like that first derived by Markin and Liberman in 1973 (Biofizika 18:453). For small applied potentials expressions are also provided for the voltage transients in charge-pulse experiments and for the membrane admittance.


Linear dichroism of rhodopsin in air-water interface films. Air-water interface films of purified cattle rhodopsin and defined phospholipids are formed by the osmotic lysis of reconstituted membrane vesicles. The interface films thus formed consist of a phospholipid monolayer containing vesicle membrane fragments. Rhodopsin molecules at the interface are restricted within the membrane fragments where they are spectrophotometrically intact and capable of undergoing photoregeneration and chemical regeneration. Multilayers of up to 8 layers can be built from these interface films. The visible absorption band of rhodopsin in these multilayers is linearly dichroic. Quantitative analysis of the linear dichroism reveals that the dipole moment of transition of the retinal chromophore in rhodopsin forms an angle of 15 degrees +/- 4 degrees with the plane of the membrane fragments in the interface film. This orientation of the chromophore relative to the plane of the membrane is essentially the same as that observed in the intact retina. Thus, the orientation of rhodopsin in the interface films is similar to that in the intact disc membranes.


Host immune responses after administration of inactivated Venezuelan equine encephalomyelitis virus vaccines--III. Kinetics for neutralizing antibody immunoglobulin class responses in donors and adoptively-immunized recipients. C57B16 mice were immunized with either live, attenuated TC-83 strain VEE virus vaccine or formalin-inactivated VEE vaccine combined with Bordetella pertussis. The kinetics of specific donor and adoptively-immunized recipient anti-VEE neutralizing antibody responses were studied. Donor mice immunized with either live or inactivated VEE virus vaccine combined with potent adjuvants develop specific anti-VEE IgM and IgG responses as early as 7 days post-immunization. Anti-VEE IgM antibody responses comprise the majority of anti-VEE neutralizing antibody at this early time period. By 14 to 21 days post-immunization, anti-VEE IgG responses predominated. When adoptively-immunized recipients were studied, the anti-VEE IgM to IgG predominance seen in donors early after administration was reversed, and for each time-period studied, recipients' serum anti-VEE antibody class responses consisted principally of IgG rather than IgM antibody. Since T-cells cooperation with B-cells is critical in the IgM-IgG antibody shift, these studies support the critical role T-cells exert in adoptive transfer in a murine model of experimental VEE infection. Furthermore, immunization with either live or inactivated VEE vaccine coupled to a potent adjuvant induce comparable donor and adoptively-immunized recipient anti-VEE antibody class responses.


Vitamin D2 in vertebrate evolution.

Vitamin D2 in vertebrate evolution. 1. The affinities of the specific vitamin D plasma transport proteins for 25-hydroxyergocalciferol and 25-hydroxycholecalciferol were studied in sixty three vertebrate species. 2. Fish, reptile, bird and monotreme plasma proteins bound 25-hydroxyergocalciferol considerably less efficiently than 25-hydroxycholecalciferol. 3. Vitamin D transport proteins from twenty-two placental mammals bound 25-hydroxyergocalciferol and 25-hydroxycholecalciferol with equal efficiency. 4. Proteins from nine mammals bound 25-hydroxycholecalciferol 10-30% more efficiently than 25-hydroxyergocalciferol.


Binding properties of serum vitamin D transport proteins in vertebrates for 24R, 25-dihydroxycholecalciferol and 24S, 25-dihydroxycholecalciferol in vitro. 1. The affinities of the specific vitamin D plasma transport proteins for 25-hydroxycholecalciferol, 24R, 25-dihydroxycholecalciferol and 24S, 25-dihydroxycholecalciferol were studied in 34 vertebrate species. 2. Fish plasma proteins bound 25-hydroxycholecalciferol, 24R, 25-dihydroxycholecalciferol and 24S, 25-dihydroxycholecalciferol with equal efficiency. 3. Vitamin D transport proteins in birds and a monotreme bound 25-hydroxycholecalciferol more efficiently than 24R, 25-dihydroxycholecalciferol; in one bird the two seco-steroids were bound with equal efficiency. 4. Transport proteins from marsupial and placental mammals bound 24R, 25-dihydroxycholecalciferol more efficiently than 24S, 25-dihydroxycholecalciferol. 5. Twelve mammal transport proteins bound 25-hydroxycholecalciferol and 24R, 25-dihydroxycholecalciferol with equal efficiency, however, in six mammals 25-hydroxycholecalciferol was more efficiently bound.


Phylogenetic and regional variations in brain gangliosides of tetrapods. 1. Ganglioside patterns were analyzed from four neural tissues (medulla, midbrain, forebrain and retina) in a representative from each of the four tetrapod classes. 2. Regional variations in ganglioside patterns were noted within some species, but differences were greater across phylogenetic lines. 3. These results suggest that evolutionary history plays a greater role than neural differentiation in the expression of brain ganglioside patterns.


Amphibian intestinal brush border membranes-I. Isolation from Rana catesbeiana tadpole. 1. Intestinal brush border membrane vesicles have been isolated form Rana catesbeiana tadpole. 2. Electron microscopy of brush border membrane vesicles demonstrates a fairly homogenous preparation of vesicles, some of them still containing electron dense material. 3. The dense vesicles probably comprise both microvillus core and membrane. 4. Negative staining of vesicles reveals the presence of knob-like structures (particles) covering the outer surface of the membrane. 5. The membranous fraction is characterized by a high specific activity of alkaline phosphatase, trehalase, glucoamylase, maltase and gamma-glutamyltranspeptidase.


Hexokinase III from Rana catesbeiana.

Hexokinase III from Rana catesbeiana. 1. Hexokinase III was partially purified from the liver of the American bullfrong, Rana catesbeiana, using DEAE-cellulose column chromatography. 2. It was inhibited by glucose concentrations above 5 x 10(-5) M (pH 5.9), 10(-4) M (pH 6.7) or 10(-3) M (pH 7.5). 3. There was virtually no inhibition by excess glucose at pH 8.7. 4. The maximum velocity of the reaction increased with increasing pH. 5. Galactose could not be utilized as a substrate. 6. Classical Michaelis-Menten kinetics were obtained with respect to ATP, with no evidence of allostery. 7. The apparent Michaelis constant for ATP was 0.23 +/- 0.013 mM in the presence of 0.2 mM glucose at pH 7.5.


Specificity of carbohydrate inhibition on leopard frog (Rana pipiens) erythrocyte agglutinating activity by concanavalin A and phytohemagglutinin. 1. Leopard frog (Rana pipiens) erythrocytes are agglutinated by the plant lectins concanavalin A and phytohemagglutinin. 2. Concanavalin A hemagglutination can be specifically inhibited by the sugars mannose, glucose, arabohexulose, alpha-methyl-D-mannopyranoside, N-acetyl-D-glucosamine, maltose, sucrose, trehalose and melezitose. 3. Sugars inhibiting concanavalin A hemagglutination are structurally composed of alpha-D-glucopyranosyl, alpha-D-mannopyranosyl or beta-D-fructofuranosyl residues with the same hydroxyl groups at carbons 3, 4 and 6. 4. Phytohemagglutinin hemagglutination can only be inhibited by the sugar N-acetyl-D-galactosamine. 5. Sugar inhibition of concanavalin A and phytohemagglutinin leopard frog erythrocyte agglutination is the same as that observed in many mammalian species.


Solubilization of frog brain and retina cholinesterase and studies of different molecular forms. 1. The cholinesterase (ChE) of frog brain and retina could be easily solubilized. About 10% of the brain and 20% of the retina ChE were found to be soluble in 0.05 M phosphate buffer. After treatment with 0.5% (v/v) Triton X-100, about 30% of the total ChE activity of the brain and only 10% for retina was left particle bound. NaCl by itself did not solubilize ChE. Use of higher NaCl concentrations in combination with Triton X-100 as well as higher detergent concentrations alone seemed to cause an inhibiting effect of the solubilized ChE from retina. 2. The solubilized ChE from brain as well as retina were electrofocused as one main activity peak, corresponding to isoelectric points of pH 6.1 and 6.0, respectively. A second molecular form at pH 5.9 was distinguishable for the brain, but not for retina ChE. 3. Sucrose gradient centrifugation indicated that the ChE solubilized from the brain and retina consists of two molecular forms exhibiting S values of 5.1 +/- 0.24, 10.9 +/- 0.33 and 6.1 +/- 0.30, 10.9 +/- 0.43, respectively. After solubilization by higher Triton X-100 concentrations the soluble extracts from brain and retina seemed to contain the activity of these forms in different proportions. 4. Polyacrylamide gel electrophoresis separated three molecular forms of the brain ChE. One of these forms was found to have a molecular weight of 394,000 +/- 20,000. The others were found to have an identical molecular weight of 550,000 +/- 10,000. Two molecular forms exhibiting molecular weights of 292,000 +/- 10,000 and 470,000 +/- 10,000, could be separated for retina.(ABSTRACT TRUNCATED AT 250 WORDS)


Folinic acid: pharmacokinetics and pharmacodynamics. In this paper I will attempt to focus attention on the uncertainties of folinic acid rescue therapy, as well as what is known.


Fracture-separation of the distal humeral epiphysis in young children. Fracture-separation of the distal humeral epiphysis in young children is frequently misdiagnosed as a fracture of the lateral humeral condyle or a dislocation of the elbow. The roentgenographic relationships between the ossification center of the capitulum humeri and the radius are important in distinguishing between the three conditions. Arthrographic studies in our patients showed that this is an extracapsular supracondylar fracture. In six fracture-separations treated either by open reduction and wire fixation or in overhead traction, the results were excellent.


Laval implant sutdies with Lucilia cuprina. Semi-field studies, using the larval implant technique, show that sheep may be protected against resistant larvae of Lucilia cuprina by thorough jetting with chlorfenvinphos, diazinon, dichlofenthion and fenthion ethyl, but not bromophos ethyl, for the normal duration of waves of bodystrike in New South Wales. The standard method of assessing implant data is to be preferred to analyses based on average protection due to the extra time required for the latter procedure.


Sampling rates required for digital recording of intracellular and extracellular cardiac potentials. Electrocardiograms and cardiac electrograms now frequently are measured for both clinical and experimental purposes by direct digital sampling, with no recording of the signal in analog form. This study examined the question of what sampling rates were required to measure accurately the continuous waveforms from the digital samples. Body surface waveforms and intracellular and extracellular waveforms measured directly from cardiac tissues were evaluated. Cardiac measurements included waveforms from the atrium, ventricle, atrioventricular transmission system and individual Purkinje strands. Sampling rates as high as 15,000 samples/sec were required to record accurately extracellular waveforms of the ventricular conduction system. Decreasing sampling rates were required as the recording site shifted through the ventricle to the body surface, where sampling rates as high as 1500 samples/sec were necessary.


Antacid and placebo produced similar pain relief in duodenal ulcer patients. The effectiveness of antacid and placebo in relieving single episodes of spontaneous duodenal ulcer pain were compared in two double blind, controlled, randomized trials. The trials compared the effects on ulcer pain of individual doses of a liquid antacid and placebo, rather than the effects of therapeutic regimens with antacid or placebo. Thirty patients were studied. There were no significant differences between antacid and placebo in time to onset, degree, or duration of pain relief. These results suggest that factors other than gastric acid neutralization are important in acute relief of spontaneous duodenal ulcer pain.


A randomized, double blind controlled trial of the efficacy of immune serum globulin for the prevention of post-transfusion hepatitis. A Veterans Administration cooperative study. A double blind, randomized, controlled trial has been conducted in 11 Veterans Administration hospitals during a 49-month period to compare the relative efficacies of immune serum globulin (ISG) and an albumin placebo for the prevention of post-transfusion hepatitis (PTH). A total of 2204 patients, of whom 1094 received ISG, participated in the study. The results indicate that ISG significantly reduced the incidence of icteric type non-B hepatitis only (inferred to be also type non-A hepatitis). Adverse reactions were rare, and the ISG did not significantly alter the incubation period or duration of the disease. The data suggest, however, that a similar reduction in type non-A, non-B hepatitis would have occurred had commercial blood been excluded from use. Analysis of the 241 patients who developed hepatitis indicates that type B hepatitis constituted less than 20% of the cases each year of the study. Furthermore, the efficacy of the ISG, manufactured in 1944, against apparent type non-A, non-B hepatitis suggests that this overlooked disease has existed from at least that time. Host- and transfusion-related factors that might have modified the development of PTH were examined. The use of commercial blood was observed to be the most important risk factor. It is concluded that the PTH incidence can be most effectively reduced by eliminating commercial donor blood, and continuing to screen volunteer donors for hepatitis B surface antigen (HBsAg) by sensitive procedures. Of prime importance is the need to define the agent(s) responsible for type non-A, non-B hepatitis.


Hepatic artery hypertrophy and sinusoidal hypertension in advanced schistosomiasis. In 5 patients with portal hypertension caused by schistosomiasis, the sinusoidal pressure (wedged hepatic pressure) varied from 20.7 to 35.4 mm Hg. While the catheter was in an occluded position within the hepatic vein and the patients were undergoing splenectomy, the main trunk of the hepatic artery was clamped. The sinusoidal pressures then fell to levels that varied from 3.7 to 7.4 mm Hg but returned to previous levels when  the clamping was released. Wedged hepatic venous pressure levels, which were  significantly greater than portal venous pressure values, decreased minimally after splenectomy. Portal venous pressure levels, however, fell to 63% of presplenectomy levels. In a control case with an enlarged spleen (cavernous hemangioma, but with a normal liver, the wedged hepatic pressure was 7.4 mm Hg and showed no alteration after clamping of hepatic artery. These data point out the importance of hepatic artery hypertrophy, that has already been demonstrated in other studies, in causing elevation of the wedged hepatic pressure in advanced hepatic schistosomiasis.


Effect of deoxycholic acid ingestion on bile acid metabolism and biliary lipid secretion in normal subjects. The effect of deoxycholate ingestion, 750 mg per day, on bile acid kinetics, biliary bile acid composition, and biliary lipid secretion was studied in 7 healthy volunteers. Bile acid kinetics were measured by isotope dilution, and hourly outputs of bile acid, cholesterol, and phospholipid were quantitated by a duodenal perfusion technique during a 24-hr period which included three liquid meals and an overnight fast. Biliary bile acid composition was assessed by coupled gas chromatography-mass spectrometry. After deoxycholic acid ingestion, biliary bile acids became composed of predominantly deoxycholyl conjugates, and deoxycholic acid pools increased 4-fold. Both chenodeoxycholic and cholic acid pools decreased, and daily synthesis of each of the primary bile acids was inhibited by 50%. Total bile acid pools did not change in any consistent manner. Daily bile acid secretion increased slightly during deoxycholic acid ingestion, and recycling frequency varied reciprocally with the total bile acid pool both before and during deoxycholic acid treatment. Deoxycholic acid ingestion caused no change in either the daily secretion of cholesterol or lecithin, or the cholesterol saturation of fasting-state bile, which remained unsaturated throughout the study. SGOT levels increased to 4 times the upper limits of normal in 2 of 7 subjects, but these levels promptly returned to normal when deoxycholate feeding was stopped. Serum cholesterol levels decreased in every subject (average 15%) during deoxycholic acid administration. No evidence for a direct role of deoxycholate in the pathogenesis of cholesterol cholelithiasis was obtained in these studies.


Pigment gallstones.

Pigment gallstones. Pigment gallstones are defined as any dark brown-to-black stone, consisting of calcium salts of bilirubin, phosphate, carbonate and other anions, and can be separated into carbonate- and noncarbonate-containing groups. Pigment stones predominate in the rural Orient, in cirrhosis, and in elderly United States patients undergoing cholecystectomy. Clinical associations include bile duct obstruction, stasis, and possibly hemolysis. Of pigment stones, 50% are radioopaque and account for two-thirds of all opaque stones. The concentrations of bile salts, phospholipids,, cholesterol, and total bilirubin in bile are similar to normal levels, but the concentration of unconjugated bilirubin is increased in the bile of some patients. Increased unconjugated bilirubin in bile may be caused by increased hydrolysis of excreted conjugated bilirubin. Unconjugated bilirubin is solubilized by bile salts, but the interaction is primarily nonmicellar. Ionized calcium and pH are important determinants of solubility. Sulfated glycoproteins, excreted in increased amounts in patients with cholelithiasis, may be the site of pigment stone precipitation because these compounds bind calcium salts tightly. E coli is frequently cultured from pigment stones in Japan but not in the United States; thus, bacterial beta-glucuronidase may be important in stone formation in Japan but probably not in the West. Stasis leads to increased calcium secretion and to increases in the concentration of sparingly soluble compounds that may then precipitate. Incomplete emptying of the gallbladder may result in the same concentration process. Unsaturated fats and chronic vagal stimulation cause pigment stone formation in animals. At present, surgery is the only treatment for pigment lithiasis.


Evidence that pancreatic proteases enhance vitamin B12 absorption by acting on curde preparations of hog gastric intrinsic factor and human gastric juice. Crude preparations of hog gastric intrinsic factor or their own previously collected gastric juices administered with labeled vitamin B12 did not enhance vitamin B12 absorption in patients with vitamin B12 malabsorption secondary to pancreatic insufficiency. However, when these sources of gastric intrinsic factor were incubated with three times crystallized preparations of insolubilized bovine trypsin or chymotrypsin, the proteolytic enzymes were removed by centrifugation, and the preparations of gastric intrinsic factor were readministered to these patients, the absorption of vitamin B12 was markedly enhanced. Studies of hog gastric intrinsic factor before and after exposure to proteolytic enzymes failed to show any difference on Sephadex chromatography or polyacrylamide gel electrophoresis or on its affinity for vitamin B12 or the ileal receptor in guinea pigs. These investigations demonstrate that: (1) gastric intrinsic factor as secreted by subjects with pancreatic insufficiency or obtained from hog pyloric mucosal extracts is ineffective in promoting vitamin B12 absorption in patients with pancreatic insufficiency, (2) incubation of crude preparations of gastric intrinsic factor with insolubilized pancreatic proteases modified these preparations of gastric intrinsic factor in an as yet undefined manner, allowing them to enhance vitamin B12 absorption, and (3) in vitro studies using gut sacs or brush border preparations do not reflect the abnormality in vitamin B12 absorption associated with pancreatic dysfunction.


Effect of graded amounts of acid instilled into the duodenum on pancreatic bicarbonate secretion and plasma secretin in duodenal ulcer patients and normal subjects. Pancreatic bicarbonate secretion and plasma secretin were measured in response to graded amounts of hydrochloric acid (0.94, 1.88, 3.75, and 7.5 mEq per 5 min) instilled directly into the duodenum in duodenal ulcer patients and in normal subjects. These graded amounts of acid produced graded increases in pancreatic bicarbonate in both groups. Mean bicarbonate secretion was significantly greater in the duodenal ulcer patients than in the normal subjects basally and after the lowest dose of hydrochloric acid. Mean (+/- SE) peak 30-min bicarbonate output (in milliequivalents) after duodenal acidification was 3.0 +/- 1.0 in the duodenal ulcer patients and 2.1 +/- 0.6 in the normal subjects (P less than 0.5). A significant (P less than 0.05) increase in plasma secretin occurred after duodenal acidification, but the increase in the duodenal ulcer and normal subjects was not significantly different (P less than 0.2). These results indicate that pancreatic bicarbonate secretion and increase in plasma secretin in response to graded amounts of duodenal acid are at least as great in patients with duodenal ulcer as in normal subjects.


Histological localization of two dipeptidases in the pig small intestine and liver, using immunofluorescence. Antibodies were raised in rabbits against glycylleucine dipeptidase (EC 3.4.13.2) and proline dipeptidase (EC 3.4.13.9) purified from the pig. IgG fractions were isolated and purified by affinity chromatography on Sepharose (dipeptidase) columns and then used in histological studies on the small intestine and the liver from pig, using the double layer immunofluorescence-staining technique. In the small intestine the dipeptidases were predominantly localized in the cytoplasm of the enterocytes. A reaction was also seen in the leukocyte cytoplasm in the stroma of the small intestine. The dipeptidases were present in the villi but not in the crypts. In the liver the dipeptidases were localized in the cytoplasm of the hepatocyte, but strong fluorescence was also obtained in the epithelium of the bile ducts. Glycylleucine dipeptidase and proline dipeptidase have been isolated from liver tissue and they were shown to be immunologically identical with the purified intestinal dipeptidases.


Antibody-forming potential of lymph nodes in aged mice, with special reference to the influence of adjuvant. The secondary antibody-forming potential of non-splenic lymphatic tissues during senescence was investigated in NMRI/Han mice, both at the cellular and humoral levels. The mean life span of conventionally reared NMRI/Han mice amounts to 19.86 months. After primary immunization of aged (20-month-old) NMRI mice with 4 X 10(8) sheep erythrocytes (SE) by the intraperitoneal (i.p.) route, the primary antibody-forming potential of both spleen and lymph nodes was significantly reduced, as compared to young adult (3-month-old) controls. In contrast, the anamnestic response elicited by an i.p. booster injection of 4 X 10(8) SE at the 44th day after primary immunization was not significantly diminished in comparison to the controls. When killed cells of Bordetella pertussis were found to be significantly increased in young adult as well as in aged mice. These data obtained at the cellular level were in accordance with corresponding serological findings. The impressive restitution of the antibody-forming potential evident after secondary antigenic stimulation was associated with a remarkable restitution of the lymph node morphology. This was particularly pronounced in the pronounced in the parathymic lymph nodes which represent the draining nodes for the peritoneal cavity. These findings indicate that the lymph nodes of the senescent individual also possess remarkable reserves in immunocompetence.


Possible molecular mechanisms of ageing. While the error theory of ageing has attracted most interest in recent times it cannot yet be regarded as being demonstrated. Posttranslational modifications of proteins genetic theory appears loical but has little in vivo evidence to prove it. Basic mechanisms of ageing probably involve the interaction of several processes.


Clinical applications of biofeedback: implications for psychiatry. The authors briefly describe biofeedback techniques such as EMG feedback, temperature feedback, and heart rate feedback, along with reports from the literature about their application to specific problems such as subvocalization while reading, Raynaud's disease, cardiac arrhythmias, and epilepsy. Many clinical applications of biofeedback are aimed at inducing relaxation, a state that has important psychotherapeutic potential. The authors suggest that biofeedback could be used to reduce a patient's general level of arousal or as an adjunct to behavior therapy or insight therapy. While there have been no reports in the literature of any harmful effects secondary to feedback training, the authors caution that some patients may respond negatively to an objectively measured state of relaxation.


Hypothalamic hamartoma: a source of luteinizing-hormone-releasing factor in precocious puberty. The presence of a hypothalamic hamartoma and precocious puberty in a 19-month-old boy provided an opportunity to study their relation. Excised tissue had the ultrastructural characteristics of an independent neuroendocrine unit -- i.e., neurons containing neurosecretory granules and blood vessels with fenestrated endothelium and double basement membranes. Immunofluorescence studies using specific antibody to luteinizing-hormone-releasing factor showed antigenicity to the factor in the hamartoma. The testicular-hypothalamic-pituitary axis was tested. Clomiphene unresponsiveness suggested a lack of maturation of central-nervous-system events characteristic of normal puberty. The negative feedback system between gonad and brain was intact but partially resistant to steroid suppression. These studies suggest that hypothalamic hamartomas may cause precocious puberty by autonomous production and release of luteinizing-hormone-releasing factor into vessels that communicate with the pituitary portal blood system.


Transperitoneal percutaneous retroperitoneal lymph node aspiration biopsy. Percutaneous aspiration biopsies of opacified retroperitoneal lymph nodes, and retroperitoneal, intraperitoneal and paraspinal masses were successfully accomplished in 14 of 17 patients. A 23-guage needle was utilized for the procedure which is performed under fluoroscopic guidance. Metastatic carcinoma, sarcoma and melanoma were readily identified by aspiration biopsy while the diagnosis of lymphoma, especially as to type, was more difficult. No significant complications have resulted from the passage of the needle through the peritoneal cavity.


Radiography of gynecomastia and other disorders of the male breast. Mammographic features of 22 pathologically proved cases reflecting disorders of the male breast are described. Two patterns of gynecomastia were observed: a dendritic pattern seen in association with breast enlargement for 6 months or more, and a more florid triangular pattern, seen in association with breast enlargement of recent onset. Male breast carcinoma may be distinguished from gynecomastia by its eccentric location, spiculation, and, in some cases, calcification or involvement of the skin and nipple. Benign conditions simulating carcinoma included a case of drug-induced gynecomastia and a case of inflamed inclusion cyst.


Automated diagnosis of congenital dislocation of the hip. A study of the automated diagnosis of luxatio coxae congenita (LCC) was performed to establish an automated screening system for hip radiographs. A specific parameter with high diagnostic accuracy was selected by the use of information theory. Pattern recognition programs were written to extract the parameter from digitized hip radiographs. Parameter measurement was performed on 10 radiographs and compared with manual measurements. Results showed good agreement with measurements obtained by specialists. This study confirms the possibilities of automated diagnosis of LCC.


The pear-shaped bladder.

The pear-shaped bladder. The tear-drop or pear-shaped bladder was originally described in cases of pelvic hematoma. It may also be seen, however, with a variety of other entities, including pelvic lipomatosis, inferior vena cava occlusion, lymphocysts, and enlarged pelvic lymph nodes. Pertinent radiographic findings of these conditions are reviewed.


The effect of respiratory carcinogenesis on systemic humoral and cell-mediated immunity of Syrian golden hamsters. The effect of intratracheal instillation of benzo(a)pyrene and its noncarcinogenic analog benzo(a)pyrene on the systemic humoral and cell-mediated immune response of Syrian golden hamsters was evaluated. Hamsters treated with the carcinogen had a transient suppression of the splenic plaque-forming cell response to sheep erythrocytes, compared with analog-treated controls. The numbers of direct (immunoglobulin M) and indirect (immunoglobulin G) plaque-forming cells were suppressed at the 9th week of treatment and then recovered to control levels. No suppression in the cell-mediated immune response, as assessed by the rejection of Chinese hamster skin grafts, was found.


Induction of gene mutations and gene conversions by vinyl chloride metabolites in yeast. Chloroethylene oxide and 2-chloroacetaldehyde, two metabolites of vinyl chloride, and 2-chloroethanol, a putative metabolic intermediate, were assayed for their genetic activity in the yeasts Schizosaccharomyces pombe and Saccharomyces cerevisiae. Chloroethylene oxide was found to be the most effective in inducing forward mutations in Sch. pombe and gene conversions in S. cerevisiae, increasing the mutation and conversion frequencies 340 and 50 times, respectively, over those of the controls. In either the presence or the absence of mouse liver microsomes, 2-chloroacetaldehyde showed only feeble genetic activity, and 2-chloroethanol was completely inactive in both yeast strains. In contrast to vinyl chloride, 2-chloroacetaldehyde did not induce forward mutations in Sch. pombe inthe host-mediated assay in mice. The results strongly support the hypothesis that chloroethylene oxide is one of the principal mutagenic agents formed from vinyl chloride in the presence of mouse liver enzymes.


Changes in distribution of human malignant melanoma membrane antigens in the presence of human antibody by immunofluorescence. Four viable human melanoma cell lines demonstrated full-surface fluorescence (FSF) after incubation at room temperature with antisera from three melanoma patients receiving autologous or homologous immunization with irradiated cultured melanoma cells and Bacillus Calmette-Guérin. By sequential immuno-fluorescent staining, it had been shown that FSF was followed by capping and extrusion of antigen-antibody complexes. One melanoma cell line was tested against its autologous antiserum. In the presence of excess antigen, FSF cells peaked to 25% of the total cell population at 30 min. Maximum capping (20%) was noted at 3 hr. When excess antibody was present, FSF cells were 30% at 2 min and capped cells peaked to 25% at 1.5 hr. Capping ceased at 4 degrees under both conditions. When the serum-treated cells were reincubated after fixation with methanol, 80 to 85% of the cells showed FSF at 2 min to 13.5 hr. Negative controls were preimmune sera and skin fibroblasts. These experimental data suggest the presence of embedded and protruding melanoma membrane antigens in the phospholipid layer. Methanol amy dissolve the lipid layer and expose the embedded antigens. The extrusion of melanoma antigen-human antibody complexes in vitro seems to be a possible mechanism of antigen shedding by melanoma cells in vivo.


Novel method for estimating the labeling index in clinical specimens with the use of immunoperoxidase-labeled. The labeling index determined by [3H]thymidine autoradiography in cells from clinical specimens was compared with the percentage of the cells showing nuclear reactivity to immunoperoxidase-labeled antinucleoside antibodies. This nuclear immunoreactivity is specific for denatured or single-stranded DNA's and is detectable almost exclusively during DNA synthesis. Results with the two methods showed excellent agreement. The new method allowed rapid accurate assessment of S phase in the tumor cells from freshly isolated aspirated specimens as well as frozen sections, suggesting general applicability to estimation of the labeling index without autoradiography.


The comparative efficacy of cephalexin and sulfisoxazole in acute urinary tract infection in children. A double blind, comparative study of the efficacy of cephalexin versus sulfisoxazole was conducted on 100 children with initial episodes of urinary tract infections. The overall bacteriologic and clinical cure rates were comparable for both antimicrobials. Children treated with cephalexin had a clinical cure rate of 86 per cent and a bacterial cure rate of 84 per cent, while those given sulfisoxazole were found to have rates of 82 and 92 per cent respectively. However, cephalexin was noted to have a rather high rate of failure in the therapy of Proteus micrabilis infections (4/8), casting some doubt on its use in urinary infections caused by the organism. Untoward effects associated with either medication were minimal.


Pulmonary function following severe acute respiratory failure and high levels of positive end-expiratory pressure. In an 18-month period, we treated 561 patients with mechanical ventilation. Fifty-four (10 percent) of these patients had acute respiratory failure, requiring treatment with positive end-expiratory pressure (PEEP) in excess of 20 mm Hg (range, 20 to 40 mm Hg). All patients were allowed to breathe spontaneously between volume-limited mechanical breaths delivered at a rate sufficient to maintain an arterial pH greater than or equal to 7.35. PEEP was applied until calculated pulmonary venous admixture was minimized. Forty-three (80 percent) of these 54 patients were alive and asymptomatic three months after dischage from the hospital, and tests of pulmonary function were performed on ten patients within one year after hospitalization. Abnormalities in pulmonary function appeared to be reversible, and pulmonary function gradually approached normal within one year. It appears that neither acute respiratory failure nor exposure to high airway pressures caused significant permanent pulmonary damage in the ten patients studied.


[Computed tomography of the pancreas (author's transl)]. The computed tomography gives direct visualisation of the pancreas in a transverse section. Form, size, and changed consistency of the organ can be diagnosed. Being a non-invasive technique it does not stress the patient, and can be applied to the severly ill with acute haemorrhagic pancreatitis. Other indications are chronic pancreatitis, pancreatic abscess, pseudo-cyst and cancer. The differential diagnosis of cancer, especially from chronic pancreatitis, may be difficult. Further methods of investigation such as arteriography or endoscopic retrograde pancreatography may also have to be used.


Sequential analysis of the releasing and fuel function of glucose in isolated perifused pancreatic islets. Isolated islets were continuously perifused with glucose to test their secretory capacity in a dynamic fashion, and were subsequently transferred to an incubation vial to measure their capacity for metabolizing glucose. Insulin release was measured by radioimmunoassay and metabolism of glucose by determining the rate of 3H2O formation from glucose tritiated on carbon atom 2 or 5 and by lactate accumulation. Insulin release was induced by glucose at a threshold of 5 mM, was half maximal at 8 mM, maximal at 15 mM and showed biphasic kinetics, which is consistent with published data. However, in contrast to most previous reports, utilization of glucose and lactate formation showed hyperbolic concentration dependency curves. Maximal and half maximal rates of glucose use were obtained at 30 and 8 mM, respectively, and lactate formation reached highest rates at 8 mM glucose. Physiological changes of glucose levels (from 5 to 10 mM) increased hormone release 4-fold (from 0.49 to 1.94 muU/islet/min) whereas glucose use was changed only slightly (from 52 to 75 pmol/islet/h), and lactate formation not at all. These data show that there is only limited association between metabolic and insulin releasing efficiency of glucose in pancreatic islets in vitro and also implicate and threshold phenomenon triggered by either a glucose metabolite or glucose itself.


Progesterone production by cultured preantral rat granulosa cells? Stimulation by androgens. Progesterone (P) production by isolated rat granulosa cells from preantral follicles was enhanced by addition of androgens to the tissue culture medium. Testosterone (T) at 10(-7), 10(-6), and 10(-4)M as well as 10(-6)M dihydrotestosterone (DHT) increased P production 400 to 700% over paired control cultures. Human chorionic gonadotropin (100 mIU/ml) and 17beta-estradiol (7.8 X 10(-10M) had no effect on P production. P was identified by both a specific radioimmunoassay and sephadex LH-20 column chromatography. The stimulatory influence of T and DHT on these preantral follicular cells is consistent with a direct role for androgens in granulosa cell differentiation.


Differential response of luteinizing hormone-releasing hormone in the basal hypothalamus and the preoptic area following anterior hypothalamic deafferentation and/or castration in male rats. Serum LH, FSH and LHRH concentrations and the LHRH content in the medial basal hypothalamus (MBH) and the preoptic area (POA) were measured by radioimmunoassay in male rats 33 days after anterior hypothalamic deafferentation (AHD) and/or castration. In castrate rats following AHD, there was a significant decrease in serum LH, FSH, and LHRH concentrations, whereas, in intact rats, serum LH was elevated in AHD over the sham AHD rats. Castration and AHD each caused a significant fall in the LHRH levels in the MBH; the decline was more pronounced in rats undergoing both castration and AHD. In contrast, deafferentation in intact and castrate rats resulted in the accumulation of LHRH activity in the POA. These studies support the suggestion that a) a substantial amount of LHRH normally found in the MBH of intact and castrate male rats originates in the rostral regions and, b) the LHRH-containing neural elements within the MBH have the competence to respond to a loss in the circulating testicular steroids.


Quantitation of human pancreatic beta-cell function by immunoassay of C-peptide in urine. Human proinsulin connecting peptide (C-peptide) was measured by immunoassay in urine from 25 normal subjects, 18 patients with diabetes mellitus, and 34 patients with various degrees of renal insufficiency. Assay validation studies showed that pancreatic C-peptide was quantitatively recovered when added to urine. Fractionation of urine by gel filtration indicated that most endogenous C-peptide eluted in fractions that corresponded to the C-peptide standard. In 34 nondiabetic subjects with normal kidney function or various renal diseases, C-peptide clearance was independent of creatinine clearance over a range of 6 to 190 ml./min. Urine C-peptide clearance (5.1 +/- 0.6 ml./min.) is greater than that of insulin (1.1 +/- 0.2 ml./min.), and the total quantity of C-peptide excreted in the urine per day represents 5 per cent of pancreatic secretion, as against only 0.1 per cent of secreted insulin. Healthy subjects excreted 36 +/- 4 mug. C-peptide per 24 hours, while this value in juvenile-onset diabetics was only 1.1 +/- 0.5 mug. Adult-onset diabetics excreted 24 +/- 7 mug./24 hr., the range overlapping the excretory rates of both normal subjects and juvenile-onset diabetics. Two insulin-requiring adult-onset diabetics showed significant beta-cell reserve during the course of acute infections. These results suggest that urine C-peptide provides a useful means of assessing beta-cell secretory capacity over a period of time and is especially advantageous when frequent blood sampling is not feasible.


Isolation and characterization of Escherichia coli K-12 F- mutants defective in conjugation with an I-type donor. Escherichia coli K-12 F- mutants defective in conjugation with an I-type donor (ConI-) were isolated and characterized. These mutants are specific in that they are conjugation proficient with other types of donor strains. They have an altered susceptibility to phages and detergents. Chemical analysis of the cell envelopes of mutant strains has shown that the lipopolysaccharide (LPS) is altered and that one major outer-membrane protein is absent. Conjugation experiments in which LPS from wild-type cells was added to a mating mixture, made up with wild-type donor and recipient cells, showed inhibition in transconjugant formation when an I-type donor, but not an F-type donor, was used. This strongly suggests that LPS of the recipient cell is directly involved in the ability to mate with an I-type donor but not with an F-type donor. The mutations are located in the 78- to 82-min region of the E. coli map, with one exception where the mutation maps near or in the galactose operon.


Biosynthesis of prodigiosin by non-proliferating wild-type Serratia marcescens and mutants deficient in catabolism of alanine, histidine, and proline. Mutants of Serratia marcescens Nima, designated as Aut, Hut, or Put, did not utilize L-alanine, L-histidine, or L-proline, respectively, as a sole carbon source but did utilize other amino acids or glycerol as carbon sources. The bacteria were permeable to alanine, histidine, and proline but lacked the enzymes responsible for degradation of these amino acids. The Aut mutant contained no L-alanine dehydrogenase activity, whereas the Hut and Put mutants contained only 7 and 4% of the histidase and proline oxidase activities, respectively, found in the wild-type strain. Rates of oxygen uptake and protein synthesis were significantly lower when the mutants were incubated in the presence of amino acids they could not degrade. Studies of L-[14C]alanine, L-[14C]histidine, and L-[14C]proline incorporation into prodigiosin synthesized by these mutants and the wild-type strain revealed that proline was incorporated intact, whereas all of alanine except the carboxyl group was incorporated into the pigment molecule. Histidine did not enter prodigiosin directly. These data suggested that the presence of unique biosynthetic pathways, independent of primary metabolism, leads to formation of prodigiosin from specific amino acids.


Effect of entry exclusion on mating aggregates and transconjugants. Mating aggregates during conjugation directed by an F-like R factor in Escherichia coli were measured as the number of Lac+-Lac- sectored colonies present in a mating mixture. There is a high degree of correlation between the concentration of transconjugants produced in a mating mixture and the concentration of mating aggregates observed at several different concentrations of donor and recipient cells. The mating aggregates are sex pilus specific as demonstrated by the ability of donor-specific ribonucleic acid phage MS-2 to decrease both mating aggregates and transconjugants in a mating mixture. During entry exclusion by either a derepressed or a repressed F-like R factor, isogenic to the superinfecting R factor except for a resistance determinant, the number of transconjugants was markedly reduced, but the number of mating aggregates was not decreased. Entry exclusion by F-Gal toward the donor HfrH resembled that of the F-like R factor in that there was a reduction in the number of recombinants but no significant decrease in mating aggregates. These results suggest that entry exclusion inhibits conjugation at a stage after the formation of mating aggregates.


Phospholipid composition and phenotypic correction of an envC division mutant of Escherichia coli. The cytoplasmic and outer membranes of a nonconditional chain-forming mutant, Escherichia coli PM61 envC, were separated by sucrose density gradient centrifugation. The phosphatidylglycerol/cardiolipin ratio in both membrane fractions was about one-third as high as in the parental strain P678. The increased level of cardiolipin in PM61 membranes is the result of an alteration of the polyglycerophosphatide cycle. It was found that the turnover rate of phosphatidylglycerol is more rapid in PM61 than in the parental strain but that its cardiolipin turnover is not significantly different. The envC mutation can be corrected phenotypically by increasing the osmolarity of the medium. In the presence of 0.6 M sucrose, the population of PM61 is composed of short rods, and the phosphatidylglycerol/cardiolipin ratio is shifted to that of the parent. The phosphatidylglycerol turns over more slowly, whereas the cardiolipin turns over more rapidly in both strains. Thus, the increase of external osmolarity acts on phospholipid metabolism as well as on an unknown step involved in the mechanism of cell division of the envC mutant.


Tetrahydrofolate-dependent biosynthesis of ribothymidine in transfer ribonucleic acids of Gram-positive bacteria. Trimethoprim, an inhibitor that prevents tetrahydrofolate-dependent transmethylation reactions inbacteria, was used in a comparative study to discriminate between two possible biosynthetic pathways, either the S-adenosylmethionine or the tetrahydrofolate-dependent formation of ribothymidine (rT) in transfer ribonucleic acids (tRNA's) of several strains of gram-positive and gram-negative microorganisms. rT-deficient tRNA's accumulate in trimethoprim-treated gram-positive Streptococcus faecium, Staphylococcus aureus, Corynebacterium bovis, Arthrobacter albidus, and all examined Bacillaceae, except Bacillus stearothermophilus. The rT-deficient rT-deficient tRNA's accept the methyl moiety from S-adenosylmethionine in vitro, with extracts from Escherichia coli (wild type) as a source of methylating enzymes; 90% of the incorporated methyl groups are present in rT. Trimethoprim does not inhibit the biosynthesis of rT in tRNA of gram-negative Enterobacteriaceae, Rhizobium lupini, and Pseudomonadaceae, suggesting that the rT-specific tRNA methyltransferases of these gram-negative strains use S-adenosylmethionine as coenzyme.


Transport and utilization of D-methionine and other methionine sources in Escherichia coli. The transport and utilization of D-methionine was investigated in several strains of Escherichia coli K-12. Wild-type cells exhibit a single transport system with a Km of 1.16 muM. This activity exhibits a specificity similar to that of the uptake of L-methionine. The activity toward the D-isomer and the high-affinity uptake of L-methionine are lost in strains mutant in metD, along with the ability to utilize D-methionine as methionine source. Both activities respond identically to gene dosage of metD and are both restored in revertants or transductants. However, although L-methionine is a potent inhibitor of D-methionine uptake, D-methionine has little or no effect on the uptake of the L-isomer. No mutants altered in the uptake of only one of the two isomers were found in a screening. Regulation of both activities was similar in their response to the internal methionine pool, and some evidence was suggestive of partial repressive control of these activities. The evidence is most consistent with the role of the metD product as a common step for two methionine-specific uptake systems, but other gene products may represent the initial substrate binding sites. This system also appears to be involved in the uptake of N-acetyl methionine and methionine sulfoxide and methionine sulfoximine. The uptake of the keto analogue of methionine, alpha-keto-gamma-methiol butyrate, appears to be mediated by a separate system specific for alpha-keto straight-chain acids 5- to 6-carbon units in length.


Role of a sugar-lipid intermediate in colanic acid synthesis by Escherichia coli. Membrane fractions from a lon strain of Escherichia coli but not a wild-type strain catalyze the incorporation of fucose from guanosine 5'-diphosphate-fucose into a lipid and into polymeric material. Both incorporation reactions specifically require only uridine 5'-diphosphate (UDP)-glucose. The sugar lipid was shown to be an intermediate in the synthesis of the polymer which was related to colanic acid. The sugar lipid had the structure (fucose3, glucose2)-glucose P-P-lipid. Its behavior on column and thin-layer chromatography, the rates of its hydrolysis in acid and base, and the response of its synthesis to inhibitors are all identical to the other sugar-lipid intermediates which have been shown to contain sugars attached to the C55-polyisoprenol, undecaprenol, by a pyrophosphate linkage. The membrane fractions from both the lon strain and the wild-type strain also catalyzed the incorporation of either glucose from UDP-glucose or galactose from UDP-galactose into a lipid fraction which was shown to contain the free sugar attached by a monophosphate linkage to an undecaprenol-like lipid. This lipid was isolated and its nuclear magnetic resonance spectra was identical to undecaprenol. The membrane fractions from both strains also incorporated glucose from UDP-glucose into glycogen and into a polymer that behaved like Escherichia coli lipopolysaccharide. Conditions were found where the incorporation of glucose could be directed specifically into each compound by adding the appropriate inhibitors.


Relation of cell growth and colicin tolerance to vitamin B12 uptake in Escherichia coli. The uptake of vitamin B12 was measured in cells of Escherichia coli whose growth had been inhibited by any of a variety of treatments. In all cases, the secondary, energy-dependent phase of B12 uptake was depressed in proportion to the decrease in growth rate, but uptake was constant in cells growing logarithmically at different rates. The depression of B12 uptake activity was independent of the site of cell metabolism affected by the inhibitor or by its effect on cell viability, and was both more rapid and of greater degree than the effects on the uptake of any of the six amino acids tested. The decline was not affected by inhibitors of either cell division or proteolysis and was manifested without any apparent decrease in the surface B12 binding activity. Transport activity was rapidly regained upon reversal of the inhibition of protein synthesis. Prompted by this response, the uptake of B12 was contrasted to the apparent uptake of the E colicins, which share the same outer membrane receptor. Sensitivity to colicin E1, measured by its inhibition of proline uptake, was not affected by growth inhibition by antibiotic treatment. Finally, there was no specific depression of B12 uptake in cells rendered colicin tolerant either by mutation or as a consequence of phage f1 infection.


Genetic inversion in the formation of an Hfr strain from a temperature-sensitive F' gal strain. An Hfr strain (PB15) that carries a duplicated copy of the galactose operon genes flanking the integrated sex factor is unusually stable since it does not show excision of the repeated deoxyribonucleic acid segment. The right-hand galactose operon is in the normal orientation. Deletion mutations that eliminate the right-hand galactose genes, the sex factor, and some of the left-hand operon have been isolated. Mutants believed to have their left-hand galactose operon inverted were able to be induced for galactose epimerase synthesis by D-fucose but did not show escape synthesis on induction of bacteriophage lambda. Ribonucleic acid specific for the galactose operon was isolated after induction of lysogenic strains presumed to carry the galactose operon in the normal and inverted orientation. Hybridization to the isolated left and right strands of lambdapgal showed that the noninformational strand of the left-hand galactose operon of the deletion mutant of PB15 was transcribed on escape induction. These results show that inversion has occurred.


Thymidine-requiring mutants of Salmonella typhimurium that are defective in deoxyuridine 5'-phosphate synthesis. In a Salmonella typhimurium strain made diploid for the thy region by introduction of the Escherichia coli episome, F'15, mutants resistant to trimethoprim in the presence of thymidine were selected. One was shown to be defective in deoxyuridine 5'-phosphate (dUMP) synthesis; it requires deoxyuridine or thymidine for growth and is sensitive to trimethoprim in the presence of deoxyuridine. Genetic studies showed that the mutant is mutated in two genes, dcd and dum, located at 70 and 18 min, respectively, on the Salmonella linkage map. The dcd gene cotransduces 95% with udk, the structural gene for uridine kinase. Both mutations are necessary to create a deoxyuridine requirement, providing evidence for the existence of two independent pathways for dUMP synthesis. Pool studies showed that a dum mutation by itself causes a small decrease in the deoxythymidine 5'-triphosphate (dTTP) pool of the cells, whereas a dcd mutation results in a much more marked decrease. The double mutant dcd dum, when incubated in the absence of deoxyuridine, contains barely detectable levels of dTTP. Enzyme analysis revealed that dcd encodes deoxycytidine 5'-triphosphate deaminase. The gene product of the dum gene has not yet been identified; it does not encode either subunit of ribonucleoside diphosphate reductase or deoxyuridine 5'-triphosphate pyrophosphatase. Mutants deleted for the dcd-udk region of the S. typhimurium chromosome were isolated.


New mini-ColE1 as a molecular cloning vehicle. A new mini-ColE1 plasmid, designated pAC105, was isolated. It has a molecular weight of 1.6 X 10(6) and carries information for its self-replication as well as information for conferring colicin E1 immunity upon its host. Furthermore, pAC105 undergoes replication in the presence of chloramphenicol even when a foreign deoxyribonucleic acid (pSC101) is inserted into its single EcoRI restriction site. Studies in minicell-producing strains demonstrate that pAC105 codes for only two or three polypeptides of low molecular weight. The advantages of using it as a molecular cloning vehicle are discussed.


Chemical measurement of steady-state levels of ten aminoacyl-transfer ribonucleic acid synthetases in Escherichia coli. Polypeptide chains of 10 aminoacyl-transfer ribonucleic acid synthetases (those for arginine, glutamine, glutamic acid, glycine, isoleucine, leucine, lysine, phenylalanine, threonine, and valine) have been identified in lysates of Escherichia coli resolved by the O'Farrell two-dimensional gel system. By labeling cells uniformly with [14C]glucose and by measuring the total amounts of these polypeptides by their radioactivity, estimations of the steady-state, molecular amounts of these enzymes were made and compared to the number of ribosomes and elongation factors in these cells. Portions of a reference culture grown on glucose and labeled with [14C]leucine or [35S]sulfate were mixed with four cultures grown in widely different media containing [3H]leucine or [3H]leucine plus [3H]isoleucine. From the isotope ratios of the total protein and of the spots containing the synthetase chains, the chemical amount of each synthetase relative to that of the reference culture was determined. The results, where comparable, show reasonable agreement with enzyme activity measurements. In general, these synthetases each exhibit a positive correlation with growth rate in unrestricted media, indicating a strong tendency for the levels of transfer ribonucleic acid, synthetases, elongation factors, and ribosomes to remain approximately, though not exactly, in balance at different growth rates.


Mitomycin C-induced expression of trpA of Salmonella typhimurium inserted into the plasmid ColE1. EcoRI endonuclease digestion of the deoxyribonucleic acid of a phi80 transducing phage carrying the entire tryptophan (trp) operon of Salmonella typhimurium (phi80 S.t.trpE-A) yielded a 4.3 X 10(6)-dalton fragment containing intact trpE, trpD, and trpC and a 3.35 X 10(6)-dalton fragment containing intact trpA. The trpA fragment inserted into EcoRI-cleaved plasmids ColE1 and CR1 was expressed regardless of its orientation of insertion. Mitomycin C, a compound that induces colicin E1 production in ColE1-containing bacteria, stimulated tryptophan synthetase alpha production in cells containing ColE1-TRPA plasmids with the trpA fragment inserted in one orientation but not the other. We conclude that in the inducible plasmids trpA can be expressed from the colicin E1 promoter.


Inhibition of TnA translocation by TnA.

Inhibition of TnA translocation by TnA. Plasmids already containing TnA showed decreased susceptibility to the translocation of a further TnA unit when compared with related plasmids that did not contain TnA. The translocation immunity imposed by TnA is exerted only on the plasmid of which it is part. It is suggested that this desensitization by a translocation unit is a general phenomenon that reduces the mutational effects of translocation.


Initiation of protein synthesis without formylation in a mutant of Escherichia coli that grows in the absence of tetrahydrofolate. Starting from a p-aminobenzoate-requiring strain of Escherichia coli (E. coli K-12 AB3292), we have isolated mutants that can grow in the absence of p-aminobenzoate (and thus tetrahydrofolate). The following lines of evidence suggest that at least one of these mutants is capable of initiating protein synthesis without formylation of methionyl-transfer ribonucleic acid (methionyl-tRNA(fMet)). (i) tRNA isolated (and charged in vivo with [(35)S]methionine) from this mutant grown in a p-aminobenzoate-free medium contained less than 0.4% of the total methionine charged to the tRNA as formylmethionine. However, when the mutant was grown in the presence of p-aminobenzoate, 40 to 50% of the total [(35)S]methionine was detected as formylmethionine. (ii) Extracts of the mutant grown in the absence of p-aminobenzoate contained no formyl-tetrahydrofolate, but such extracts did contain formylatable methionyl-tRNA and a functional transformylase. (iii) Tetrahydrofolate-free extracts of the mutant were capable of supporting protein synthesis with viral RNA (from f2) as messenger, but the resulting synthesized proteins contained no formylmethionine, and methionine residues were detected where formylmethionine residues are normally found. In the presence of formyl-tetrahydrofolate, use of a similar extract resulted in the detection of 30 to 40% of the total polypeptide methionine as formylmethionine. (iv) Initiation of protein synthesis in vitro occurred more readily with formyl-tetrahydrofolate-free extracts of the mutant than with similar extracts prepared from the parent strain. However, in the presence of formyl-tetrahydrofolate, initiation of protein synthesis proceeded equally well with both kinds of extracts. tRNA from this mutant and another spontaneously derived mutant was found to be partially deficient in the modified nucleoside ribothymidine (rT). Analysis of extracts showed that the mutants contained decreased levels of the methylase that results in the formation of ribothymidine. In vivo studies with an independently isolated rT(-) strain suggest that the lack of rT in tRNA facilitates the growth of E. coli under conditions where protein synthesis is forced to take place without formylation.


Size and turnover of polyadenylic acid-containing ribonucleic acids in a fragile mutant of Saccharomyces cerevisiae. Ribonucleic acid-containing polyadenylic acid [poly(A)+-RNA] was studied in lysates from an osmotic-sensitive mutant of Saccharomyces cerevisiae characterized by low nuclease activity. The poly(A)+-RNA fraction, analyzed by electrophoresis in polyacrylamide-formamide gels, constitutes a heterogeneous population of molecules, with molecular weights ranging from 0.2 X 10(6) to 3 X 10(6) and having an average of 1.2 X 10(6). The turnover rate of poly(A)+-RNA was determined by the decay of radioactivity after a cold uracil chase, and the observed half-life of 21 min corresponds to about 10% of the cell doubling time. Poly(A)+-RNA was analyzed by gel electrophoresis under denaturing and non-denaturing conditions. A correlation was established between the apparent secondary structure and the turnover rate of poly(A)+-RNA species.


Deoxyribonucleic acid sequence organization of a yeast plasmid. Two-micrometer deoxyribonucleic acid (DNA), a circular plasmid of Saccharomyces cerevisiae, contains two nontandem repeated sequences which are inverted with respect to one another. These repeated sequences together account for 21% of the molecule length. Restriction endonuclease analysis and electron microscopy demonstrated the existence of two forms of 2-mum DNA differing in the orientation of the interstitial segments bounded by the inverted repeated sequences. The two forms of 2-mum DNA could result from an intramolecular reciprocal recombination between inverted repeat elements. A map containing the restriction endonuclease sites and the units of the inverted repeat has been constructed.


Isolation, mapping, and examination of effects of TnA insertions in ColE1 plasmids. Twelve TnA insertions of ColE1 deoxyribonucleic acid have been isolated and mapped by electron microscopic studies of heteroduplex molecules. Insertions only blocking the production of active colicin clustered in one region of the map, whereas insertions only blocking the expression of deoxyribonucleic acid nicking activity associated with the plasmid "relaxation complex" clustered in another region of the map. The location of one insertion that blocks both the expression of colicin immunity and the production of active colicin suggests that the expression of both characteristics are coordinately controlled.


Fine-structure mapping of the rts, rplK, rplL, and rpoB genes of Escherichia coli. The specialized transducing phage lambdacI857S7drifd18 was used as a donor in a transductional mapping of four genes in the rif region of the Escherichia coli genome. The gene order was rts-2.9-rplL-0.8-rpoB, where the numbers indicate intermarker distances in kilobases. The possible orientation of these genes with respect to each other and to neighboring genes is discussed.


Number of mutations required to evolve a new lactase function in Escherichia coli. The frequency of mutation of the ebgAo allele to ebgA+ was compared with the frequency of mutation of strA+ to strA-. The observation that both spontaneous and ethyl methane sulfonate-induced mutations to ebgA+ occurred more frequently than mutations to strA- suggests that ebgA+ mutants arise as the result of single-point mutations.


Threonyl-transfer ribonucleic acid synthetase and the regulation of the threonine operon in Escherichia coli. Two threonine-requiring mutants with derepressed expression of the threonine operon were isolated from an Escherichia coli K-12 strain containing two copies of the thr operon. One of them carries a leaky mutation in ilvA (the structural gene for threonine deaminase), which creates an isoleucine limitation and therefore derepression of the thr operon. In the second mutant, the enzymes of the thr operon were not repressed by threonine plus isoleucine; the threonyl-transfer ribonucleic acid(tRNA) synthetase from this mutant shows an apparent Km for threonine 200-fold higher than that of the parental strain. The gene, called thrS, coding for threonyl-tRNA synthetase was located around 30 min on the E. coli map. The regulatory properties of this mutant imply the involvement of charged threonyl-tRNA or threonyl-tRNA synthetase in the regulation of the thr operon.


Fractionation of Saccharomyces cerevisiae cell populations by centrifugal elutriation. An exponential population of Saccharomyces cerevisiae cells was fractionated by centrifugal elutriation, using water as the elutriating liquid. Evidence that the population had been fractionated according to age in the cell cycle was obtained by examining the fractions for their size distribution, their microscopic appearance after Giemsa staining, and their ability to initiate synchronous growth.


Antinuclear antibody with distinct specificity for polymyositis. In the course of studying antinuclear antibodies in the rheumatic diseases, a new precipitin reaction (provisionally referred to as PM-1) was observed between calf thymus nuclear extract and polymyositis sera. Objectives of this study were to further define the immunologic nature of this reaction and to determine its specificity for polymyositis. Immunodiffusion studies using calf thymus nuclear extract revealed the PM-1 precipitin line in 17 of 28 patients with polymyositis. This reaction was not produced by sera of 460 patients with other diseases. Enzyme and heat treatments of the nuclear extract showed that PM-1 was distinct from native DNA, ribonucleoprotein, and Sm antigens. Fractionation of PM-1-positive serum by 30% ammonium sulphate and Sephadex G-200 chromatography revealed that the factor producing the PM-1 precipitin reaction was in a serum fraction which showed only IgG by immunoelectrphoresis against anti-whole human serum. Because of the apparent strong specificity, the PM-1 system may represent a marker antibody for polymyositis.


Properties of two nickel-chromium crown-and-bridge alloys for porcelain veneering. Composition, microstructure, mechanical properties, and alloy-porcelain bond strength of two proprietary nickel-chromium-based crown-and-bridge alloys were studied. The materials exhibited compositional as well as microstructural differences. Wiron-S was significantly stronger and harder than Microbond-NP. Microbond-NP yielded alloy-porcelain bond strength values comparable to those of precious alloy-porce-lain combinations.


Monocyte function in man.

Monocyte function in man. The monocyte-macrophage cell line is an important member of the host defense system. This report describes a series of assays that can be applied routinely in the evaluation of human monocyte function and gives information as to the activity of normal monocytes in these systems. Tests were chosen to assess various aspects of monocyte function that give some insight into the host defense status and the degree of "activation" of the monocyte. The assays outlined in this report are relatively simple to perform and include measurements of human monocyte chemotaxis, phagocytosis, fungal and bacterial killing, adhesion, and spreading.


Acute lymphoblastic leukemia (ALL) antigens detected with antisera to E rosette-froming and non-E rosette-forming ALL blasts. Based on the presence or absence of erythrocyte receptors(E) a T cell marker, acute lymphocytic leukemia (ALL), can be divided into E+ALL and E-ALL. We studied cell surface antigens on blasts from 12 children with untreated ALL: eight with E-ALL and four with E+ALL. Heterologous antisera were raised against thymus cells, E+ and E-ALL blasts, appropriately absorbed and tested by immunofluorescence and a radiolabeled antibody assay with normal and leukemic lymphoid cells. By both methods, anti-thymus and anti-E+ALL sera reacted with human thymocytes. Specific binding of anti-E+ALL serum to T antigens was indicated by the fact that a single absorption with thymocytes abolished its binding to allogenic thymocytes, and the reactivity of anti-E+ALL serum with thymus, blood and bone marrow lymphocytes was similar to that of anti-thymus serum. After exhaustive absorption with blood leukocytes, anti-E+ALL and E-ALL sera were negative against normal lymphocytes and bone marrow cells from children with ALL in remission. Anti-thymus and anti-E+ALL sera reacted with blasts from patients with E+ALL, but not with E-ALL. In contrast, anti-E+ALL serum reacted with 40 to 96% of blasts from all children with E-ALL, whereas of the four patients with E+ALL, two were negative and two had the lowest percentage of immunofluorescent cells (10 to 22%). These results were confirmed with the radiolabeled antibody assay. Patients with active E-ALL had cells bearing E-ALL antigen(s) in the peripheral blood and bone marrow, but the number of immunofluorescent cells was lower in blood. Cells reactive with anti-E-ALL serum did not react with thymus cells, blood lymphocytes, remission bone marrow cells, Raji cells, PWM and PHA-induced blasts and CLL cells bearing mIg (uk). These data suggest that the antigen detected on E-ALL blasts by anti-E-ALL serum is neither a HLA-related nor a cell differentiation antigen. Thus, by using antiserum to E+ALL blasts, we have confirmed the presence of a T cell-specific antigen(s) on E+ALL cells. This antiserum did not recognize other leukemia-associated antigens common to E+ and E-ALL. We have also demonstrated an antigen(s) which is regularly expressed on E-ALL blasts and is either not detectable or is present in a lower proportion of E+ALL blasts.


Activation of the classical and properdin pathways of complement by bacterial lipopolysaccharides (LPS). Bacterial lipopolysaccharides (LPS) have been demonstrated to activate both the classical and the properdin pathways of complement. The lipid A region of the LPS is responsible for classical pathway activation and the polysaccharide region responsible for properdin pathway activation. Classical pathway activation by lipid A does not depend upon antibody to the lipid A and properdin pathway activation proceeds by a lipid A-independent mechanism. The polysaccharide portion of the LPS molecule exerts a modifying influence on the potential anticomplementary activity of the lipid A.


Clinical study of a patient with lupus vulgaris before and after injection of dialyzable transfer factor. This report describes the clinical improvement and acquisition of tuberculin skin-test sensitivity by a tuberculin-negative, drug-resistant patient with lupus vulgaris after a single injection of dialyzable transfer factor (TFd) from a tuberculin-positive healthy donor. The patient's lymphocytes showed a slight response to tuberculin in the leukocyte migration inhibition test and in the lymphocyte transformation test before TFd injection. The acquisition of cellular immunity to tuberculin was demonstrated in vitro by enhanced tuberculin-induced blast transformation. A good correlation between skin test and in vitro tuberculin sensitivity and clinical improvement was seen during the three years that the patient was observed.


Separation of epidermis from dermis with sodium thiocyanate. After human skin is treated with 2 N sodium thiocyanate, epidermis is easily separated from dermis. The level of cleavage occurs at the lamina lucida of the basement membrane zone. Bullous pemphigoid antigen remains attached to the epidermis.


Immunopathology of cicatricial pemphigoid: studies of complement deposition. Immunopathologic investigations were conducted on the sera and oral mucosal tissue specimens of 23 patients with cicatricial pemphigoid. A linear, continuous basement membrane zone pattern was noted in 83% of oral mucosal biopsy specimens studied. This pattern is indistinguishable from the pattern noted in immunofluorescence studies of bullous pemphigoid, herpes gestationis, and some cases of desquamative gingivitis. Complement studies provided data supportive of classical pathway activation in cicatricial pemphigoid tissue. Deposition of IgA with Factor B, properdin, and C3 raised the possibility of alternative pathway activation, a question requiring further study. Circulating antibasement membrane zone antibodies were noted in the sera of two patients with cicatricial pemphigoid.


Alteration of effectiveness of antibiotics by anaerobiosis. In vitro antibiotic susceptibility tests of facultative organisms are routinely performed under aerobic conditions, despite the fact that many infections caused by these organisms occur in anaerobic areas, i.e., intra-abdominal absess. Experiments were performed aerobically and anaerobically to determine the susceptibility of E. coli, P. mirabilis, and K. pneumoniae to gentamicin, tobramycin, cephalothin, cefazolin, and cefamandole. Antibiotic sensitivities were determined by disc and agar dilution techniques in air and in anaerobic jars with CO2 absorbed. Tube dilution studies were performed in air and anaerobically and time-kill studies were done in aerobic and anaerobic broth. The amount of aminoglycoside required to inhibit bacterial growth was increased 4 to 20 times by anaerobiosis in 20 of 25 strains tested. Time-kill curves showed that bacterial killing by aminoglycosides was markedly impaired by anaerobiosis. Anaerobic conditions had no effect on the rate or extent of killing by cephalosporins. These data may have significance in determination of antibiotic susceptibility of facultative organisms under anaerobic tissue conditions. Antibiotic sensitivity testing done on these organisms in air may not reflect the actual state of antibiotic-bacterial interaction under conditions of the infection.


Verbal auditory hallucinations: mind, self, and society. Verbal auditory hallucinations of schizophrenic patients are usually sensed as coming from m-ltiple voices. The voices are usually not recognized by patients; they are anonymous. They are expressed mainly in the second person (as if directed to the patient by others), occasionally in the third person (as if two or more persons are overheard talking about the patient), very rarely in the first person. They are sensed as separate from the self and out of the patient's control. All of these characteristics can be said to describe how a person generalizes from his social experiences a societal evaluation of himself. In addition, the voices of schizophrenic patients are predominantly disparaging, call approbrious names, or are accusatory. Schizophrenics tend to come from and be of low socioeconomic status. The hallucinations appear to be related to the schizophrenic's sense of society's disparagement of him because of his low socioeconomic status and achievement.


Stereotaxic clipping of arterial aneurysms and arteriovenous malformations. In carefully selected cases of arterial aneurysms and deep-seated arteriovenous malformations (AVM), when direct attack may be dangerous or impossible, the authors advocate stereotaxic clipping. A special device and technique for its application are described. The instrument is introduced through a trephine opening and clipping is monitored by angiography. Successful results have been obtained in 10 operations performed on eight patients, three of whom had arterial aneurysms (two internal carotid and one anterior cerebral-anterior communicating) and five with AVM's.


Jugular--dural sinuses--jugular reflux in dynamic brain-flow imaging as a sign of unilateral innominate vein obstruction: case report. Unilateral innominate vein obstruction with patency of the superior vena cava was suspected when early jugular--sinuses--jugular reflux of tracer occurred during brain-flow imaging. Radiographic venography confirmed this pattern of venous obstruction.


The effect of the stimulant drugs, dextroamphetamine and methylphenidate, on secretion of growth hormone in hyperactive children. The stimulant effect of L-dopa (125 to 500 mg) was compared to dextroamphetamine and methylphenidate, 15, and 20 mg, respectively, on growth hormone secretion in 20 hyperactive children. All three stimulants were responsible for peak GH concentration in serum at 60 minutes after drug ingestion; there was no significant difference between the mean GH level at any time of sampling. Seven of the children were retested with L-dopa and dextroamphetamine after six to eight months of treatment with methylphenidate. After treatment, there was a tendency to higher zero time levels of GH, and to delayed and/or paradoxical response to dextroamphetamine. The findings indicate an acute and a probably long-term effect of dextroamphetamine and methylphenidate on the homeostasis of growth hormone. The possible long-term adverse effects of these drugs on the growth of children indicates the need for caution to the widespread use of these agents.


Influence of the heat treatment of human milk on some of its protective constituents. Human milk was subjected to heat treatments of graded severity and examined for its content of immunoglobulins, lactoferrin, lysozyme, vitamin B12-and folate-binder proteins, and lactoperoxidase. Holder pasteurization (62.5degrees C 30 minutes) reduced the IgA titer by 20%, and destroyed the small content of IgM and most of the lactoferrin. Lysozyme was stable to this treatment, but with an increase in temperature there was progressive destruction, to near 100% at 100degrees C. The same was broadly true of the capacity of milk to bind folic acid and potect it against bacterial uptake; with vitamin B12 the binder was more labile at 75degrees C than at 100degrees C. The milk contained no detectable lactoperoxidase.


Outbreak of meningitis in a newborn intensive care unit caused by a single Escherichia coli K1 serotype. Three cases of meningitis that occurred during a nine-day period in a newborn intensive care unit were caused by a single E. coli serotype 07:K1:H-. A single organism outbreak was suspected when the three spinal fluid isolates all possessed the same two unusual bacteriologic and biochemical characteristics: nonmotile and ornithine negative. Culture surveillance identified eight infants colonized with the same strain of E. coli; three of these infants are described. Clusters of cases of E. coli meningitis in newborn intensive care units should be evaluated and managed as potential outbreaks.


Hereditary deficiency of the third component of complement in a child with fever, skin rash, and arthralgias: response to transfusion of whole blood. A previously well 34-month-old male presenting with fever, skin rash, and arthralgias was found to lack C3 by immunochemical (undetectable) and hemolytic (1% normal) assays. No infectious agent could be demonstrated. Protein levels of Clq. C4, C5, properdin, and C3b-INA and hemolytic activities of complement components C1 to C9 except C3 were normal or elevated; total hemolytic complement activity was 13% of normal and was reconstituted by purified C3. Properdin factor B was 702 (normal 175 to 275) mug/ml, and was not cleaver upon addition of zymosan or cobra venom factor. The serum had normal immune adherence activity, but was deficient in ability to opsonize Candida albicans for uptake and Escherichia coli for killing by neurophils, generate neutrophil chemotactic factors and inhibit the growth of E. coli; these activities were restored by purified C3. A transfusion of 320 ml 1-hour-old normal whole blood on the fifty-second day resulted in transitory elevation of the C3 level to 25 mg/dl with a fall-off (approximately 2 1/2% per hour) to undetectable levels by 69 hours; it was followed by disappearance of the skin rash and arthralgias and return to normal of the previously elevated temperature and CRP levels. C3 levels in family members (seven of 24 half-normal), lack of anti-C3 activity, normal C3b-INA levels and a normal rate of catabolism of transfused C3 indicated that the deficiency was inherited with autosomal codominance and involved decreased synthesis of C3. Thus, this child is a unique individual with inherited C3 deficiency presenting with absence of repeated infections, whose symptoms of fever, skin rash, and arthralgia were abated by whole blood transfusion.


Treatment of high-renin hypertension with propranolol in children after renal transplantation. Ten children with hypertension poorly controlled with other drugs and high peripheral plasma renin activity after renal transplantation were treated with propranolol. The mean systolic pressure decreased from 139 to 127 mm Hg (p less than 0.05) and the mean diastolic pressure from 98 to 83 mm Hg (p less than 0.01). Eight children had an antihypertensive response; two did not respond. The maximum dose of propranolol in responders varied from 1.0 to 6.2 mg/kg/day and duration of treatment until response varied from four to 49 days. PRA, repeated in seven responders, decreased in all (p less than 0.01). There was no correlation between changes in PRA and blood pressure. Propranolol was well tolerated and was a valuable antihypertensive drug in these children.


An evaluation of terminal hinge position and neuromuscular position in edentulous patients. Part II. Duplicate mandibular dentures. The purpose of Part II of this study was to determine whether or not a denture fabricated to the neuromuscular position (NMP) could be as functional as one fabricated to the terminal hinge position (THP). Two mandibular dentures were fabricated for each of six edentulous subjects, one to the THP and one to the NMP, along with only one maxillary denture each. The subjects had no prior knowledge of the two mandibular dentures in order to determine the clinical significance of a difference in the two positions. These results were evaluated by clinical methods. It was concluded that the NMP is clinically acceptable for intercuspation of artificial teeth on complete dentures.


A theoretical analysis of the mechanics of the Thompson dowel semiprecision intracoronal retainer. An analysis has been made of the mechanics of the Thompson dowel retainer. The analysis locates the center of rotation during function and identifies the factors that affect its position. The degree to which the dowel should be relieved to permit unrestricted rotation has been established. The rationale of an occlusal bevel and the limits of its positioning have been analyzed. The importance of aligning the shelves of the dowel wells in two planes is emphasized and ways of handling the problem of disparity of shelf heights are given.


Anterior fixed partial dentures utilizing the acid-etch technique and a cast metal framework. A technique has been described which permits the fabrication and attachment of an anterior fixed partial denture without tooth preparation. The fixed partial denture is attached to the lingual surfaces of the abutment teeth utilizing a composite resin and acid-etched enamel. Conservation of tooth structure and minimal chair time and patient expense are the primary advantages of this technique.


A simplified method for constructing a core following endodontic treatment. This report described use of a core form. The principal objectives of the method are (1) the provision of a form of proper size and (2) to allow formation of the core portion of the dowel and core directly in the mouth in much less time than has heretofore been needed.


The pin-amalgam restoration. Part 1. A review. A review of the literature indicates that the use of pins to support amalgam or cast gold restorations in advanced caries in posterior teeth is a quick and relatively inexpensive method of restoring teeth otherwise doomed to extraction. However, the use of pins demands a careful and skillful dentist who understands the morphology of these teeth and the proper use of the materials if injury to the pulp, accidental perforations, crazing of the dentin, and splitting of teeth are to be prevented. Until more objective and controlled studies are completed, the use of cemented pins vs. self-threading pins remains a matter of empirical and personal preference. It is clear that success in the use of any pin or pins depends more on the dentist's skill and judgment than on the particular type of pin technique used. In order to more objectively evaluate the use of pin-amalgam restorations, Part 11 of this study will analyze the results obtained in the Clinic at Tufts University School of Dental Medicine.


Changes in physical properties of joined gypsum fragments. (1) The dimensional change in joined fragments of gypsum models was small. The highest percentage occurred with zinc phosphate cement. (2) The cyanoacrylate cements produced the strongest bonds. Mendent Formula VI and the epoxy cement were least effective. (3) The bond strength was generally greater after 24 hours. The exceptions were Mendent Formula VI and zinc phosphate cement. (4) Different gypsum products responded differently to bonding agents. Some are superior for plaster of Paris; others for artificial dental stone.


The use of wax and self-curing acrylic resin in border molding. A procedure for border molding of impression trays with border wax and cold-curing acrylic resin has been described. The use of modeling compound, the water bath, and the alcohol torch has been eliminated.


The surgical approach to urological complications in renal allotransplant recipients. Urinary extravasation or ureteral obstruction occurred in 22 patients who received 30 transplants in a series of 290 renal transplants. This incidence represent 10.3 per cent of the entire transplant experience at The Johns Hopkins Hospital and Baltimore City Hospitals from 1968 to the present time. Ureteroneocystostomy was used as the primary form of urinary tract reconstruction in all but 1 patient who had urinary complications. These 22 patients received 30 renal transplants: 6 from living related donors and 24 from cadaver sources. There were 15 instances of urinary extravasation and 14 instances of obstruction. All but 2 fistulas were diagnosed within 30 days of the original transplant. Obstruction occurred later, with 4 cases of ureterovesical obstruction being diagnosed 3 to 5 years after the transplant procedure. The ureterovesical junction or bladder was the site of complication in 17 of the 29 instances. Surgical management in these cases was highly individualized, with successful outcomes more commonly attained in those cases characterized by obstruction. Ureteral stents were used in all but 1 secondary procedure involving the ureter and these stents were not associated with an increased incidence of urinary tract infection. Death directly related to the urological complications occurred in 2 cases, 5 patients underwent transplant nephrectomy and 2 patients died of rejection and infection more than 6 months after the urinary fistulas were successfully managed. From the original series, there are 15 of the 22 patients who have stable renal function after secondary or tertiary urological procedures on the transplanted kidney. Four patients underwent surgical correction of hydronephrosis associated with infection or diminishing renal function more than 3 years after the transplantation and 3 of these had good results.


Fibrosis of a renal transplant ureter.

Fibrosis of a renal transplant ureter. Ureteral obstruction following renal transplantation, although not a common occurrence, is a serious complication because of the single functioning kidney. Obstruction may be caused by ureterovesical stenosis, retroperitoneal fibrosis or adhesions, clot formation, pelvic lymphoceles or kinking of the ureter. A case is presented in which there was progressive partial ureteral obstruction caused by fibrosis and stricture of the transplant ureter, which were probably owing to rejection episodes.


Gray scale nephrosonography: current status. Gray scale ultrasound evaluation of the kidney permits visualization of the texture of renal and hepatic parenchyma. This fact has led to more specific diagnoses in renal disease by non-invasive techniques. Current applications of gray scale ultrasound are presented. Ultrasound serves as a significant complementary technique to other modalities and is useful in some situations as a primary study.