Piperacillin, a new penicillin active against many bacteria resistant to other penicillins. The in vitro activity of piperacillin, a new semisynthetic piperazine penicillin derivative, was evaluated against 626 clinical isolates and compared with the activity of other beta-lactam antibiotics. At a concentration of 0.1 microgram/ml, piperacillin inhibited all streptococci except enterococci. Non-beta-lactamase-producing staphylococci were inhibited by 1.6 microgram or less per ml. Both beta-lactamase- and non-beta-lactamase-producing Haemophilus were inhibited by 0.1 microgram/ml. Piperacillin inhibited non-beta-lactamase-producing Escherichia coli, Salmonella, and Shigella at a concentration of 6.3 micrograms/ml, but 20% of strains of these species containing type III beta-lactamase were not inhibited by 100 micrograms/ml. Piperacillin at 25 micrograms/ml, inhibited 83% of Citrobacter, 58% of Klebsiella, 88% of Enterobacter, and 50% of indole-positive Proteus, Acinetobacter, and Providencia. At 25 micrograms/ml, piperacillin inhibited 95% of Pseudomonas aeruginosa and 78% of Bacteroides fragilis. The minimal inhibitory concentration of piperacillin against Pseudomonas was affected by increasing the inoculum size and by pH. Minimum bactericidal concentrations against Pseudomonas and Serratia often were eightfold greater than the minimum inhibitory concentrations. Piperacillin was equal in activity to ampicillin against enterococci. It was more active than carbenicillin against E. coli, Klebsiella, Enterobacter, and Bacteroides. It was the most active penicillin against Pseudomonas and inhibited many strains of Pseudomonas for which the MICs of carbenicillin were above 200 micrograms/ml. Piperacillin was hydrolyzed by many different beta-lactamases. Synergistic activity of piperacillin was demonstrated when it was combined with amikacin, gentamicin, and cefazolin against P. aeruginosa and members of the Enterobacteriaceae. No antagonism was observed when piperacillin was combined with aminoglycosides; however, antagonism was observed rarely against E. coli when piperacillin was combined with cefazolin.


In vitro study of clavulanic acid in combination with penicillin, amoxycillin, and carbenicillin. The activity of clavulanic acid alone and in combination with penicillin, amoxycillin, and carbenicillin was studied. Marked reductions in the minimum inhibitory concentrations (MICs) for a wide spectrum of beta-lactamase-producing clinical isolates were found. Of particular interest were the decreased MICs of penicillin for Bacteroides fragilis and beta-lactamase-producing strains of Neisseria gonorrhoea in the presence of the clavulanic acid. Beta-lactamase-producing strains of Escherichia coli, Klebsiella spp., and indole-negative Proteus also showed considerably increased susceptibility to amoxycillin in combination with clavulanic acid. Two beta-lactamase-producing strains of Pseudomonas aeruginosa remained resistant to carbenicillin in the presence of clavulanic acid.


Totally synthetic medium for susceptibility testing. The results of disk diffusion and plate dilution susceptibility testing of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Proteus mirabilis on media gelled with agar-agar or with a synthetic hydrogel were compared. Synthetic hydrogel can be combined with a totally defined synthetic amino acid medium to yield a reproducible, totally defined, synthetic solid medium without the antagonistic or booster effects of some components of agar. Such a medium could be used as a reference medium for susceptibility testing.


Penicillin-netilmicin synergism against Streptococcus faecalis. The combination of penicillin plus netilmicin was synergistic in vitro against 28 strains of Streptococcus faecalis and compared favorably with penicillin in combination with gentamicin. Similarly, penicillin plus netilmicin was as effective as penicillin plus gentamicin in the therapy of 67 rabbits with enterococcal endocarditis produced with a streptomycin-susceptible (S) or a streptomycin-resistant (R) strain of S. faecalis. After 5 days of infection, control rabbits had bacterial titers of 10(10) colony-forming units (CFU)/g of vegetation. Those treated with penicillin plus netilmicin had mean titers of 10(5.2) and 10(5.1) CFU/g for S and R strains, respectively, and those treated with penicillin plus gentamicin had mean valve titers of 10(5.8) CFU/g for both strains. After 10 days of therapy, mean valve titers with penicillin plus netilmicin were 10(3.8) and 10(4.7) CFU/g, and with penicillin plus gentamicin they were 10(4.5) and 10(5.4) CFU/g for S and R strains, respectively. Thus, if netilmicin proves to be less toxic than other aminoglycoside antibiotics, it may have potential usefulness in the therapy of enterococcal endocarditis.


Influence of R-plasmid RP1 of Pseudomonas aeruginosa on cell wall composition, drug resistance, and sensitivity to cold shock. R-plasmid RP1 was transferred to Pseudomonas aeruginosa cells, as indicated by their resistance to carbenicillin, ampicillin, cephaloridine, kanamycin, and tetracycline, and by the presence of a periplasmic beta-lactamase. The wild-type cells (RP1-) were lysed by ethylenediaminetetraacetic acid but not by ethylene-glycol-bis(2-aminoethyl ether)-N,N-tetraacetic acid, whereas cells carrying the plasmid (RP1+) were resistant to both these chelating agents. RP1+ and RP1- strains were both sensitive to the lytic action of polymyxin B and the lethal action of cold shock, but the effect was less marked in the RP1+ cultures. A proportion of the RP1+ cells surviving cold shock lost resistance to carbenicillin, tetracycline, and kanamycin. The chemical composition of whole cells and cell walls of RP1+ differed from that RP1- in the content of cation, phospholipid, and markers for lipopolysaccharide and peptidoglycan. Differences in cell wall composition, response to ethylenediaminetetraacetic acid and polymyxin B, and the effects of cold shock are all compatible with the hypothesis that RP1 confers changes in the cell envelope, probably in the outer membrane, of P. aeruginosa.


Susceptibility of cephalothin-resistant gram-negative bacilli to piperacillin, cefuroxime, and other selected antibiotics. The in vitro antibacterial activity of piperacillin and cefuroxime against 180 isolates of cephalothin-resistant Enterobacteriaceae and of piperacillin against 46 isolates of Pseudomonas aeruginosa was determined. Amikacin, gentamicin, carbenicillin, cefoxitin, and cefamandole were included for comparison. The activities of piperacillin and carbenicillin against Enterobacteriaceae were comparable. Piperacillin was appreciably more active against Pseudomonas than carbenicillin and was equivalent in activity to amikacin on a weight basis. The following beta-lactam agents were the most active against the indicated organisms (in parentheses): cefoxitin (indole-positive Proteus spp.), cefuroxime and cefoxitin, (Klebsiella spp.), piperacillin (Enterobacter spp.), cefuroxime and cefoxitin (E. coli), piperacillin and cefoxitin (Serratia spp.), and cefoxitin (Providencia spp.). Amikacin inhibited 98% of Enterobacteriaceae at clinically achievable serum levels.


Susceptibility of lipopolysaccharide-defective mutants of Pseudomonas aeruginosa strain PAO to dyes, detergents, and antibiotics. Lipopolysaccharide-defective mutants of Pseudomonas aeruginosa strain PAO have been isolated on the basis of their resistance to lipopolysaccharide-specific bacteriophages. These mutants have been differentiated by their agglutination in NaCl and acriflavine, phage sensitivity, and chemical analysis of the lipopolysaccharides. The susceptibility of the wild-type strain and four mutants to a series of twenty-six agents, including dyes, detergents, antibiotics, and lysozyme, was examined. The roughest mutant (AK-43) exhibited increased susceptibility to sodium deoxycholate, hexadecylpyridinium chloride, benzalkonium chloride, ampicillin, penicillin G, erythromycin, colymycin, and polymyxin B. The role of cell envelope fractions in antibiotic resistance in P. aeruginosa is discussed.


Penicillin and netilmicin in treatment of experimental enterococcal endocarditis. Successful therapy of enterococcal endocarditis requires the use of a combination of penicillin plus an aminoglycoside. The effectiveness of penicillin (Pen), streptomycin (Str), and netilmicin (Net), a new aminoglycoside, alone and in combination, were studied in vitro and in the treatment of left-sided enterococcal endocarditis in rabbits. In vitro Pen+Str or Net resulted in a more rapid and more complete bactericidal effect than Pen, Str, or Net alone against a Str-susceptible strain of enterococcus (strain 1). Against a highly Str-resistant strain (strain 2), Pen+Net showed an advantage over Pen, Str, or Net alone, or Pen+Str. Endocarditis was produced in rabbits with strain 1 or 2, and treatment was initiated 24 h later. Rabbits were treated for 48 h or 5 days with procaine Pen, Pen+Str, or Pen+Net. With strain 1, numbers of enterococci in the vegetations decreased more rapidly with Pen+Str or Pen+Net treatment than with Pen, Str, or Net alone. With strain 2, Pen+Net showed a clear advantage over Pen, Str, Net, or Pen+Str. Net in combination with Pen showed synergistic in vitro activity and was more effective than Pen alone in the treatment of enterococcal endocarditis in rabbits caused by both Str-susceptible and Str-resistant strains.


Effect of benzylpenicillin on the synthesis and structure of the cell envelope of Neisseria gonorrhoeae. The effect of benzylpenicillin on the synthesis and morphology of the cell envelope of Neisseria gonorrhoeae was examined. Penicillin immediately stopped murein synthesis; it also enhanced the rate of turnover of glucosamine, but not diaminopimelic acid, in the murein. In addition, penicillin greatly increased the shedding of lipid and lipopolysaccharide into the medium. In the electron microscope, protrusions of the cell membrane were evident, as well as apparent holes in the murein cell wall. All of these changes occurred while active synthesis was taking place, before the lysis of the cells. Lysis could be prevented by growing the cells at low pH and high concentrations of Mg2+; however, the effects of penicillin on murein synthesis and turnover and on the release of lipid were not affected.


Comparative in vitro activities of SCE-129, sulbenicillin, gentamicin, and dibekacin against Pseudomonas. Against sulbenicillin- and gentamicin-susceptible strains of Pseudomonas aeruginosa, SCE-129 was about 10 times more active than sulbenicillin and had a similar activity to gentamicin and dibekacin. Sulbenicillin-resistant strains of P. aeruginosa were moderately resistant to SCE-129, whether these strains were gentamicin-resistant or not. Gentamicin-resistant strains of P. aeruginosa were resistant to dibekacin but not to SCE-129. Against P. maltophilia, the minimum inhibitory concentration of SCE-129 resembled those of sulbenicillin, gentamicin, and dibekacin. Most strains of P. cepacia were moderately resistant to SCE-129 and sulbenicillin and highly resistant to gentamicin and dibekacin.


Ultrastructural morphometric investigations on human platelets after long-term ingestion of oral contraceptives. Platelets from healthy females, who were taking oral contraceptives over a long period of time, were investigated by ultrastructural morphometry. The size, shape, relative volumes, numerical and surface densities of the platelets and their organelles were measured. In 'long-term pill users' the platelets showed a significant increase in the area of section profile. Further changes as, for example, increased volume of platelet granules, reduced number of mitochondria, decreased volume and surface of the open canalicular system, and increased surface of the dense tubular system are to be noted.


The gonadotropin-resistant ovary syndrome in association with secondary amenorrhea. A young patient with secondary amenorrhea and primary infertility is described. After a gynecological-endocrinological exploration including laparoscopy, the diagnosis indicates secondary hypergonadotropic, hypo-estrogenic normo-androgenic amenorrhea. The anatomopathological examination of an ovarian biopsy revealed an intact follicular apparatus, thus disproving the suspected diagnosis of climacterium praecox. Since very high gonadotropin doses could not induce an ovulation, it was concluded that the rare combination of secondary amenorrhea and the gonadotropin-resistant ovary syndrome must be present. The pathogenesis of this syndrome is discussed.


Electromyographic assessment of central motor disorders. Experimental procedures are described that were designed to assess central motor disorders quantitatively. Initially delineated is a study of triggered ballistic movement performed in a reaction-time situation. The reciprocal triphasic EMG pattern recorded in an antagonistic muscle pair was clearly abnormal in Parkinsonian patients; increased duration, diminished synchronization of motor units, and a tendency for coactivation of agonist and antagonist muscles and for action tremor were observed. Transport time of elbow movement was prolonged, particularly in a choice reaction-time situation. Rapid, passive displacements of the forearm combined with excitability measurements of hindlimb motoneurons in monkeys revealed the existence of a transcortical loop that may contribute to increased muscle tone in Parkinsonian patients.


Correlation between movement of concanavalin A membrane receptors and cytolysis. A scanning electron microscopy study. The present study was undertaken to test whether cytolysis induced by Concanavalin A (Con A) requires lateral mobility of membranal lectin receptor sites into caps. Treatment of interphase murine mastocytoma cells with 10(-4) M colchicine promoted cap formation by Con A in about 30% of the cells, followed by cytolysis. Pretreatment of the cells with NaN3, low temperature, or glutaraldehyde decreased the degree of capping and, to the same extent, the degree of cytolysis. The addition of antibodies to cells bound with Con A increased the appearance of capping and cytolysis. A linear relationship with a high correlation coefficient exists between the degree of capping and cytolysis, suggesting that lateral mobility of membrane Con A receptors is required for cytolysis by the lectin. The process of cap formation by Con A up to the stage of cytolysis was followed by scanning electron microscopy.


Differentiation of food vacuolar membranes during endocytosis in Tetrahymena. The origin and differentiation of Tetrahymena pyriformis food vacuolar membranes has been studied by freeze-fracture electron microscopy. By measuring the temperature needed to induce the onset of lipid phase separation (as inferred by the appearance of particle-free regions in replicas) and calculating the changes in average intramembrane particle distribution, a distinct modification of the vacuolar membrane could be observed from the time of its formation from disk-shaped vesicles to its maturation before egestion of its indigestible contents. Whereas the nascent vacuolar membrane first showed signs of phase separation at 9 degrees C, this temperature rose to 14 degrees C in the completed vacuole and then, after lysosomal fusion, eventually declined to 12 degrees C. The average membrane particle density on the PF face increased from 761 +/- 219 to 1,625 +/- 350 per micron 2 during membrane differentiation. Like other membranes of the cell, the vacuolar membrane underwent adaptive changes in its physical properties in cells maintained for several hours at low temperature. This exposure to low temperature caused an equal effect in vacuoles formed before, during, or after the temperature shift-down. Normal changes in the properties of the vacuolar membrane may have some bearing on its programmed sequence of fusion reactions.


Instrumental cardiovascular conditioning: a review. This paper reviews experiments, hypotheses, and current controversies about instrumental cardiovascular conditioning. Demonstrations of such conditioning in curarized animals challenged a differentiation between instrumental and classical learning on the basis of their respective effector systems but did not prove direct operant learning by the autonomic nervous system. In humans, ethical prohibition of curarization and lack of adequate controls for respiration and muscle tension have resulted in incomplete understanding of the roles of voluntary, somatic mediators. Despite a variety of potential clinical applications of biofeedback, the available literature lacks studies of its efficacy compared to more standard modes of therapy. The physiological mechanisms and central neural pathways involved in instrumental cardiovascular conditioning remain almost totally unknown.


Treatment of postanoxic intention myoclonus with valproic acid. Valproic acid in therapeutic doses was used in the treatment of postanoxic intention myoclonus. Disappearance of the myoclonus occurred with marked improvement in the electroencephalogram. No significant side effects were noted. Hepatic function tests were monitored. Determination of valproic acid plasma levels was used to guide therapy. Levels above 55 micrograms were generally required. The patient remains free of myoclonus after four and one half months.


Vitamin E deficiency and seizures in animals and man. Study of 100 children with grand mal convulsive disorders and 100 medically healthy children of matching age showed significantly lower plasma levels of vitamin E in the former (means 632.2 +/- 17.3 and 822.5 +/- 21.8 micrograms/dl respectively; p less than 0.001). This finding accords with the ability to prevent seizures in rodents by giving alpha-tocopherol before exposing them to a convulsion-inducing environment.


Degenerative progressive myoclonic epilepsy electrokymographic observations. Clinical and pathological findings in two cases of degenerative progressive myoclonic epilepsy (PME) are described. The clinically difficult task of differentiating a "cerebellar" tremor from an action myoclonus is emphasized. Simultaneous electroencephalography and electrokymography was done, using capacity to ground transients for recording hand movements. This method was found useful in corroborating the cerebellar nature of the remaining disorder, after successful treatment of the myoclonic element with anticonvulsants.


Modification of erythrocyte physicochemical properties by millimolar concentrations of glutaraldehyde. The effects of low levels of glutaraldehyde uptake (less than 120 mumol/10(10) cells) on the physicochemical properties of human red blood cells (RBC) were investigated. Salient effects include: by different measures of cell deformability, the extent of glutaraldehyde uptake required to decrease cellular deformability was shown to range from approximately 8 to 30 mumol/10(10) cells; osmotically stressed red cells exhibit complete hemolysis when the level of glutaraldehyde uptake is less than 28 mumol/10(10) cells and no hemolysis when uptake is less than 70 mumol/10(10) cells with the extent of hemolysis decreasing in an approximately linear manner with glutaraldehyde uptake between these limits; glutaraldehyde uptake of up to 58 mumol/10(10) cells does not change the cells' density, mean cell volume or ability to retain potassium.


Kinetics of glutaraldehyde fixation of erythrocytes: size, deformability, form, osmotic and hemolytic properties. Red blood cells interact with glutaraldehyde (GA) in a complex kinetic pattern of events. At a given GA concentration in phosphate buffered saline (PBS), the sequence of cell 'volume' response, as measured by resistive pulse spectroscopy (RPS), includes: an immediate response to the overall solution osmolality; a constant volume, latent phase; a rapid swelling phase; an intermediate constant volume phase; and a shrinkage phase to a final steady state volume. The final volume depends on fixative solution osmolality; for GA concentrations between 0.05% and 0.25% w/v, fixative osmolalities of less than 355 mosM, including 'isotonic', or greater than 355 mosM, lead to final cell volumes greater or less than native, respectively. Cell-membrane deformability decreases continuously and monotonically with time, as assessed by RPS. The rate of fixation is a direct function of GA concentration, in accordance with a derived empirical expression. The measured kinetic responses are related to considerations of cell size, deformability, and form, and to mechanisms involved in abrupt osmotic hemolysis.


Effects of toxicants on the somatosensory system. A computerized system for an objective and accurate study of vibration sensitivity has been designed. Sensitivity can be assessed in human as well as nonhuman primates. Its usefulness in the study of peripheral nerve disorders induced by chemical exposure is emphasized.


Effects of hyaluronidase, trypsin, and EDTA on surface composition and topography during detachment of cells in culture. Cultured human embryo fibroblasts (HLM18) were labeled with [3H]glucosamine and Na35SO4, and then treated with testicular hyaluronidase, trypsin, or EDTA. Macromolecular material from the surface of these cells was characterized by DEAE-cellulose chromatography and cetylpyridinium chloride precipitation while the associated morphology of cell detachment was studied by phase contrast and scanning electron microscopy. Release of surface glycosaminoglycans by testicular hyaluronidase did not cause cell rounding or detachment. EDTA did not release cell-surface components, but caused cell contraction and detachment morphologically similar to that caused by trypsin. Large amounts of cell-surface glycoproteins and glycosaminoglycans were released by trypsin. From these observations it is concluded that hyaluronic acid is not a principal adhesive agent in the attachment of cells to a substrate. It is suggested that both EDTA and trypsin may have their primary effect upon the cytoskeleton.


Effect of puromycin on synthesis, processing, and nucleocytoplasmic translocation of rRNA in Tetrahymena pyriformis. 1. Treatment of Tetrahymena pyriformis with various concentrations of puromycin results in a more pronounced inhibition of [3H]uridine accumulation in stable RNA than of protein synthesis. 2. At a concentration of 500 micrograms/ml, which is almost completely inhibitory to [3H]uridine incorporation in vivo, puromycin has no influence on the incorporation of [3H]UTP into RNA in isolated macronuclei. Pretreatment of the cells with the antibiotic, however, reduces the activity of RNA polymerases in isolated nuclei to less than 30%. 3. In puromycin-treated cells a small amount of pre-rRNA is synthesized but not processed into cytoplasmic rRNAs. 4. Puromycin reduces the nucleocytoplasmic translocation of pre-existing RNA to about 25% of the control rate within 5 min, resulting in an accumulation of relatively stable rRNA precursor molecules in the macronucleus.


Evolution of lipoproteins deduced from protein sequence data. 1. Human serum apolipoprotein A-I contains a prominent 11-residue sequence periodicity. 2. Similar 11-residue segments occur in the other sequenced human apolipoproteins, C-I, C-III, and A-II. 3. Computer analyses of the sequences support the hypothesis that they evolved from a common ancestor. 4. An evolutionary history of these proteins is proposed. 5. The estimated rate of change of these proteins indicates that all four types will be found throughout the vertebrates and that related proteins will also be found in invertebrates.


The temperature dependence of State IV respiration, the calcium uptake system, and the activity of the calcium ionophore A23187 in mitochondria from endo- and ectothermic animals. 1. Arrhenius plots of State IV respiratory activity of liver mitochondria from both rainbow trout and rat were linear over the temperature range 5-35 degrees C. 2. Calcium uptake was monitored by stimulation of oxygen consumption and by calcium electrode recording, with quite comparable results. Rainbow trout gave the usual linear Arrhenius plot but this plot for rat mitochondria exhibited two well-defined inflections or discontinuities. 3. The temperature dependence of the activity of the ionophore A23187 was investigated by measuring the increase in oxygen uptake following a sub-maximal dose of this drug. Again a linear relation was found for rainbow trout, but in this case the rat curves showed only a single inflection point. 4. These results are discussed in relation to other work on the effects of lipid phase transitions on mitochondrial membrane-associated systems.


A progress report on the status of a new disease of American cats: cytauxzoonosis. A protozoon disease of domestic cats in the U.S.A. which resembled diseases of African ungulates caused by blood parasites of the family Theileridae and the genus Cytauxzoon was first described by scientists of the University of Missouri in 1976. The Plum Island Animal Disease Center of the U.S. Department of Agriculture (USDA) became involved in an effort to discover the relationship of the parasite to those of African species and to ascertain if the disease posed a threat to American livestock. More than 500 cats were infected and the disease studied experimentally by scientists of both institutions. Passage of the parasite was accomplished by parenteral administration of fresh or deep frozen blood or tissues from infected cats. With the exception of a single cat, immunized at PIADC, the experimental disease has terminated fatally about 20 days following exposure. Most cats died within 9-15 days after infection. Diagnosis of the disease is accomplished by study of blood smears, stained with Giemsa and by examination of sections from major visceral organs. Piroplasms are not always found in blood smears, but large masses of schizonts are always found in at least one of the following organs: lungs, spleen, liver or lymph nodes. Since the disease almost invariably has a fatal termination in cats, it is believed that the reservoir is in another species of animal. The new disease has now been reported from Missouri, Arkansas, Texas and Georgia, with new isolations of the parasite having been made in recent months.


Glycogen metabolism in the snail, Biomphalaria glabrata. 1. The distribution of glycogen in different tissues of the snail, Biomphalaria glabrata was determined; the cephalopedal region contained 15-17 mg, the mantle region 23-29 mg, the hepatopancreas 31-45 mg, and the ovotestis 50 mg of glycogen/g wet wt. 2. There was a significant decrease in the glycogen content in the cephalopedal region after incubating the snails with 5-hydroxytryptamine (4 x 10(-5) M) in vivo. 3. Enzymatic degradation of glycogen from different tissues revealed that the degree of branching was 9% of the total glucosyl residues, and the length of outer branches was about 40% of the total glucosyl residues. 4. There was an active form of glycogen phosphorylase in the cephalopedal region and the mantle region. Phosphorylase in the hepatopancreas was generally inactive, but could be activated by an endogenous phosphorylase kinase. 5. Glycogen synthetase of the snail tissues required glucose-6-phosphate to be active.


Glycogen synthase in the rat tapeworm, Hymenolepis diminuta--I. Enzyme activity during development and with crowding. 1. Activity of glycogen synthase (E.C. 2.4.1.11) in Hymenolepis diminuta (Cestoda: Cyclophyllidea) was investigated as a function of development and with crowding. 2. Synthase activity was low in the anterior and posterior ends of the worms and highest in the pregravid proglottids in the mid-portion of the strobila. 3. The enzyme activity increased during development of the cestode at least up to 15 days postinfection, but the increase in activity apparently was not due to conversion of the inactive to the active form. 4. Mature oncospheres also contained glycogen synthase, but the activity was lower than in strobilar tissues. 5. Synthase I activities and the proportion of total activity in the I form were generally higher in worms from high density (100 worm) infections than in those from low density (10 worm) infections.


Glycogen synthase in the rat tapeworm, Hymenolepis diminuta--II. Control of enzyme activity by glucose and glycogen. 1. The proportion of activity in the physiologically active I form of glycogen synthase in Hymenolepis diminuta (Cestoda) decreased in the worm when the rat host was fasted and was greatly increased in the cestode 1 hr after a 24 hr fasted rat was refed. 2. The increase in glycogen synthase I activity was due to glucose present in the host gut after feeding, not to other physiological changes in the rat intestine due to meal consumption. 3. Incubation of intact H. diminuta in vitro with glucose also resulted in the conversion of glycogen synthase D to I. 4. Glucose does not appear to affect the glycogen synthase complex directly, because neither the total synthase converted to I nor the rate of conversion was affected by glucose in a partially purified homogenate. 5. High concentrations of glycogen inhibited the synthase D to I conversion and high mol. wt glycogen was a more effective inhibitor than low mol. wt glycogen.


Hormonal regulation of trehalose metabolism in the blowfly Phormia Regina Meig.: effects of cardiacectomy and allatectomy at the subcellular level. 1. Kinetic properties of adult Phormia fat body glycogen synthetase were studied and compared to other animals. The KM for UDPG is 2.82 mM, decreasing to 0.58 mM in the presence of G-6-P. 2. The specific activity of fat body glycogen synthetase shows a reduction of 30% within 2 days after allatectomy. 3. Fat body T-6-P synthetase activity decreases to 70% of the control value after cardiacectomy. 4. Corpus cardiacum homogenate fails to induce higher T-6-P synthetase activity in cell-free preparations from cardiacectomized flies. 5. Interactions between corpus cardiacum and corpus allatum in regulating carbohydrate metabolism are discussed.


Apolipoproteins and lipoprotein families in chicken embryos and egg yolk. 1. Turkey anti-LP-A and anti-LP-B cross react strongly with whole adult chicken serum. 2. Similar reactions occur with whole chicken embryo sera (10-21 days of incubation) and egg yolk fluid. 3. Ultracentrifugation of sera from 14-day old chicken embryos, after reaction with anti-LP-B, reveal complete precipitation of the VLDL and LDL of embryo serum (indicating that adult-type LP-B is the major apolipoprotein of these density classes). 4. All of the chicken embryo adult-type apo-LP-A is in the HDL fraction. 5. However, egg yolk and hen serum VLDL contain apo-LP-A as well as apo-LP-B.


Immunochemical identity of the high and low molecular weight forms of Galapagos marine iguana hemoglobin. 1. Two forms of Galapagos marine iguana methemoglobin, with molecular weights of 140,000 and 70,000 daltons, were identified in iguana RBC lysates by Sephadex G-200 molecular sieve fractionation. 2. The 140,000 dalton ferric hemoglobin was isolated by DEAE-Sephadex A-50 ion-exchange chromatography and found to be pure by electrophoretic and immunological criteria. 3. Immunochemical analyses revealed the high and low molecular weight hemoglobins to be antigenically identical.


Frog liver glycogen synthase. In vitro and in vivo interconversions between I and D forms. 1. Frog liver has enzymatic systems able to interconvert glycogen synthase. 2. D to I conversion is achieved in vitro by incubation at 30 degrees C. ATP, ADP, inorganic phosphate and glycogen are inhibitors of this conversion, whereas glucose-6-P and Mg2+ stimulate it. 3. I to D conversion in vitro depends on ATP-Mg2+. Cyclic-AMP activates this conversion, while glucose-6-P inhibits it. 4. Injection of glucose, ribose, mannose, fructose, galactose, and cortisone into frogs increase liver percentage of I activity. 5. Glucagon and adrenaline decrease percentage of I activity.


The effects of concanavalin A, cerulenin, hydroxyurea and tunicamycin on the incorporation of amino acids and glucosamine in Tetrahymena pyriformis. 1. Pulse labelling of conjugating Tetrahymena pyriformis with glycine and glucosamine gave results consistent with a requirement for de novo synthesis of a lipid-like component which is involved in the addition of sugar to cell glycoprotein. 2. Conjugation of complementary strains of Tetrahymena can be inhibited without gross morphologic and metabolic changes in the cells. The observed inhibition of conjugation is reversible. 3. Concanavalin A stopped conjugation and stimulated both glycine and glucosamine incorporation. 4. Tunicamycin inhibited conjugation but had little influence on total glucosamine incorporation. Amino acid incorporation was slightly enhanced. 5. Hydroxyurea blocked conjugation, inhibited the incorporation of glucosamine and seemed to induce a transient increase in glycine incorporation. 6. Cerulenin inhibited conjugation but had little effect on the incorporation of glycine and glucosamine into the cell.


Synchronizing temperatures and rRNA metabolism in Tetrahymena pyriformis. 1. In Tetrahymena pyriformis pre-rRNA is synthesized and efficiently processed and translocated into the cytoplasm at both a supraoptimal (34 degrees C) and a suboptimal (8 degrees C) synchronizing temperature. 2. At the heat shock temperature (34 degrees C) no substantial differences in the kinetics of the main intracellular events of rRNA-metabolism compared to the optimal growth temperature (28 degrees C) were found. 3. The high temperature, however, induces a strong retardation of uptake of [3H]adenosine into the cells and a reduction of the cellular ATP pool size. 4. At the cold shock temperature (8 degrees C) the rates of transcription and nucleocytoplasmic transport of rRNA as well as of nucleotide pool equilibration are reduced to a similar extent (25-30% of the optimal rates at 28 degrees C). 5. The results are discussed and compared with the effects of sub- and supraoptimal temperatures on rRNA synthesis and processing in mammalian cells.


Purine nucleoside phosphorylase from bovine liver. 1. Purine nucleoside phosphorylase (purine nucleoside:orthophosphate ribosyltransferase, E.C. 2.4.2.1) from liver of cattle, Bos taurus, was purified to homogeneity. Some properties of the enzymes from three different bovine tissues were compared and discussed. 2. The enzyme has a molecular weight of 83,000, a sedimentation coefficient of 5.3 S, a Stokes' radius of 3.71 nm, a frictional ratio of 1.30 and a subunit molecular weight of 30,000. 3. Optimal pH for xanthosine degradation is around 5.5, whereas a broad pH activity profile for inosine degradation was observed between 5.0 and 7.5. Lineweaver-Burk plots curved downward at high concentrations of substrates, inosine, phosphate and arsenate.


Comparative specificities of trehalases from various species. 1. Using derivatives or non-symmetrical analogs of alpha,alpha-trehalose, we studied the catalytic specificities of trehalases from various species: Pseudomonas fluorescens, Melolontha vulgaris, porcine and human kidneys. 2. alpha,Beta-trehalose, beta,beta-trehalose, 6,6'dideoxy alpha,alpha-trehalose, alpha-D-xylopyranosyl alpha-D-xylopyranoside were shown to be neither substrates nor inhibitors. 3. 6'deoxy alpha,alpha-trehalose, alpha-D-glucopyranosyl alpha-D-xylopyranoside, alpha-D-allopyranosyl alpha-D-glucopyranoside and alpha-D-galactosyl alpha-D-glucopyranoside, which all possess an intact alpha-D-glucopyranosyl residue, were split by all these trehalases. 4. alpha-D-glucopyranosyl alpha-D-mannopyranoside, alpha,alpha-trehalosamine are competitive inhibitors. 5. These results show the importance of the primary alcohol group at C-6, of the equatorial configuration of the OH groups at C-2, C-3 and C-4 and of the modification of the structure at C-2 of the substrate for the catalytic activity.


Activity of adenosine deaminase and purine nucleoside phosphorylase in erythrocytes and lymphocytes of man, horse and cattle. 1. Activities of ADA and PNP were measured in erythrocytes and lymphocytes of man, horse and cattle. 2. In bovine hemolysates both enzyme activities are low when compared with activities in human hemolysates. In horse hemolysates both enzyme activities are virtually absent. 3. Enzyme activities are consistently lower (about 50%) in intact lymphocytes than in sonicated lymphocytes. This finding suggests that the uptake of nucleosides is rate-limiting for both enzymes in intact lymphocytes. 4. The activity of ADA in horse lymphocytes is comparable to that in lymphocytes of patients with severe combined immunodeficiency associated with ADA deficiency.


Partial characterization of pepsins and gastricsins and their zymogens from human and toad gastric mucosae. 1. Three zymogens have been isolated from human gastric mucosae and two from the stomachs of the toad Caudiverbera caudiverbera. 2. Human zymogens I and III were immunologically related and cross-reacted with antisera prepared against porcine pepsinogen. The third, (II), showed no cross-reactivity. 3. Human zymogens I and III and toad zymogen ZII gave rise to two human pepsins and to a pepsin-like enzyme, respectively. 4. Human zymogen II (gastricsinogen) and toad zymogen ZI gave rise to human gastricsin and to a gastricsin-like enzyme respectively. 5. The toad enzymes showed much greater stability at neutral and alkaline pH values than the human enzymes.


Plasma carnitine and cholesterol levels in monkeys and pigeons: species/breed differences and the influence of gender and diet. 1. Total plasma carnitine and cholesterol were studied in two breeds of pigeon and four species of sub-human primates. 2. The levels of plasma carnitine and cholesterol differed within species and between species and were influenced by diet and gender. 3. In the animals studied, pigeons had the highest levels of plasma carnitine (greater than 120 nM/ml) and Squirrel monkeys (Saimiri sciureus) had the lowest levels (ca. 10 nM/ml). 4. Linear regression analysis of plasma carnitine and cholesterol in the monkeys and pigeons indicated no strong correlation between these parameters.


Toad carbonic anhydrase: purification of the enzyme from erythrocytes of Bufo marinus and comparison with the enzyme activity in the urinary bladder. 1. Two forms of carbonic anhydrase, having isoelectric points of 6.1 and 5.8, were purified from erythrocytes of the toad, Bufo marinus, and the presence of a third form, pI = 5.4, was demonstrated. 2. Each of the two purified isozymes catalyzed the hydration of CO2 and the hydrolysis of nitrophenyl acetate esters at rates characteristic of Type C (or high-activity) forms of carbonic anhydrase. 3. Both forms of the erythrocyte enzyme have similar molecular weights (approx 29,000), amino acid composition, sensitivity to acetazolamide, and kinetic properties. 4. The epithelium of the toad's urinary bladder also was found to contain significant amounts of carbonic anhydrase, which appears by isoelectric focusing to be indistinguishable from the enzyme isolated from the erythrocyte.


Precipitation of animal plasma proteins by puff-adder venom. 1. Plasma and serum samples obtained from various animals never previously exposed to snakes or snake venom were diffused against different concentrations of puff-adder, Bitis arietans, venom using the double immunodiffusion technique. 2. Depending upon venom concentration, two precipitin arcs could be produced in the case of all plasma samples used. No serum samples showed any arcs except pigeon serum, where one precipitin line was observed. 3. By altering the concentration of snake venom between 1% and 10% when immunodiffusing against plasma a change in position of the precipitin lines was observed and also the disappearance of one or both of the two bands at higher concentrations. This indicates that the arcs observed are in all probability due to precipitation of plasma protein fractions. 4. Previous results indicated that one of the two bands observed when diffusing venom against plasma was due to the precipitation of fibrinogen. By diffusing snake venom against heparin we have now shown that the second band involves this molecule and is not due to another coagulation factor as was suggested previously.


Purification and partial characterization of high and low activity carbonic anhydrase isoenzymes from Malaclemys terrapin centrata. 1. High activity (CA C) and low activity (CA B) carbonic anhydrase isoenzymes have been purified from turtle erythrocytes. 2. The two isoenzymes differed in CO2 hydration specific activity by 36-fold. 3. The low activity isoenzyme contained one half-cystine residue, whereas the high activity isoenzyme contained four half-cystines and required a reducing environment to maintain activity. Both isoenzymes contained zinc. 4. Molecular weights of 28,500 and 30,400 daltons were established for the low and high activity isoenzymes respectively. 5. Both isoenzymes were inhibited by acetazolamide, but only the high activity isoenzyme was inhibited by parachloromercuribenzoate. 6. The low activity isoenzyme was present in the erythrocytes at about 8-10 times the concentration of the high activity isoenzyme. 7. The high activity isoenzyme cross-reacted with antibodies prepared against pure chicken carbonic anhydrase C.


Alaskan malamute chondrodysplasia--VIII. Incorporation of [14C]glucosamine and [3H]serine in hepatic metal-binding proteins of Canis familaris. 1. Hepatic proteins isolated from control kennel dogs bound small quantities of zinc and iron and the peptide fraction contained neither metal. 2. Zince loading of kennel dogs stimulated an hepatic uptake of five times more zinc and three times more iron than an equivalent copper load. The increase in metal concentration was noted in the 10,000 dalton protein. 3. Both the 12,000 and 10,000 dalton proteins isolated from kennel dogs contained more binding sites specific for zinc than for either copper or iron. All three proteins isolated from Alaskan Malamutes showed a smaller affinity for zinc than copper or iron. 4. Both copper and zinc loading stimulated an uptake of [14C]glucosamine and [3H]serine from the peptide fraction of control kennel dogs into the 10,000 dalton protein.


A comparative electrophoretic analysis of mammalian carbonic anhydrase isozymes: evidence for a third isozyme in red skeletal muscles. 1. Starch gel electrophoretic patterns of carbonic anhydrase (CA) isozymes were examined from tissue extracts of cats, sheep, rabbits and mice. 2. In addition to the widely distributed and extensively studied B and C isozymes, an additional isozyme (called CA-A) was observed. 3. Tissue distribution studies showed the A isozyme to be predominantly localized in red skeletal muscles, although this activity was also observed in white and "mixed" skeletal muscles of the cat, sheep and rabbit, as well as sheep lung and rabbit liver. 4. A, B and C isozymes of carbonic anhydrase from cat, sheep and mice exhibited independent variations in nett surface charge. In terms of decreasing anodal migration, the following results are reported: cat A greater than C greater than B; sheep C greater than B greater than A; and mouse B greater than C greater than A. 5. These results are consistent with the existence of 3 genetic loci encoding carbonic anhydrase in mammalian tissues.


Histones from gonads of the star-fish Asterias rubens. 1. Histones were isolated from gonads of the star-fish Asterias rubens and characterized by their amino acid composition and their electrophoretic migration. 2. Comparative studies with calf thymus homologous histones show the highly conservative structure of the histones H3 and H4, and the variability of the other histones namely H1 and H2B.


Gluconeogenesis in vertebrate livers.

Gluconeogenesis in vertebrate livers. 1. The hypothesis is advanced that it would be logical for a tissue (liver) to evolve as a gluconeogenic organ in order to recover the lactate produced as a result of rapid and sustained contraction of skeletal muscle. 2. Lactate was present in skeletal muscle of all animals examined and increased following electrical stimulation. It was also present in the blood. 3. Gluconeogenesis from lactate occurred in liver slices of all animals excepting amphibia. However, livers of these animals also contained much glycogen and are probably gluconeogenic. 4. Phosphoenolpyruvate carboxykinase was present in all animals investigated; pyruvate carboxylase was present in all animals excepting the toad.


Chondroitin sulfates of the epiphysial cartilages of different mammals. 1. The distribution chondroitin 4- and 6-sulfates in the epiphysial cartilages of several mammals are reported. 2. Chondroitin 6-sulfate is present in higher relative proportion in articular surfaces of young and adult epiphysial cartilages in most of the mammals studied. 3. Exception to this was found in some species of the order Rodentia in which chondroitin 4-sulfate was almost the only chondroitin present in young and adult cartilages. 4. These and other results suggest that chondroitin 4-sulfate may be an important component for the calcification process, whereas chondroitin 6-sulfate seems to be related to the integrity of the articular surfaces.


Glycolytic end products of the adult dog heartworm, Dirofilaria immitis. 1. Adult dog heartworms remained alive and motile for 24 hr without oxygen present and with only glucose available as a substrate. 2. Lactate accounted for 55% of the carbon from the 1-14C-glucose utilized in 1 hr and 14CO2 for 1.9%. 3. Only traces of 14C were found in glycogen and no net accumulation of acetate was demonstrated. 4. Dirofilaria immitis resembles Litomosoides carinii in the percent of utilized glucose appearing as lactate but is more akin to Brugia pahangi and Dipetalonema viteae in survival under anaerobic conditions and in negligible acetate production.


Aerobic and anaerobic metabolism of the crystalline lens of a poikilotherm; the toad Bufo marinus. 1. The O2 consumption and lactic acid production of the lens of the toad Bufo marinus was measured under various conditions. The energy consumption was very low compared to other amphibian tissues and about 20% of that previously described in the rat lens. About 80% was derived from oxidative metabolism, which is the converse of that seen in mammals. 2. Cyanide abolished O2 consumption and increased lactate production 20-30 times ("Pasteur effect"). 3. Under aerobic conditions about 40% of the energy requirements of the lens are related to the presence of Na in the bathing-media but this was not seen in the presence of CN. 4. Oxidative metabolism was predominant in the outer cortical regions of the lens while glycolysis persisted even in the central nucleus. The energetic requirements of this region were, however, only about 10% of those in the intact lens. 5. Lactate readily leaves the lens and passes into its bathing fluids, and at high rates of glycolysis this occurs more readily across the posterior surface. 6. The results are discussed in relation to the unique physiological needs, structure and situation of the lens.


Esterase-7, a common constituent of numerous mouse tissues. 1. Electrophoretic and staining techniques for the relatively specific demonstration of esterase-7 in mouse tissues are described. Esterase-7 was found to be a common constituent of most mouse tissues. 2. The electrophoretic polymorphisms determined by three alleles a, b and c were studied in nine mouse strains. 3. The banding pattern of esterase-7 is being described and the isoelectric points of the allelic bands are being estimated. 4. Recombination studies on a further 129 animals confirmed that the Es-7 locus is tightly linked to Es-2 and Es-11. 5. Esterase-7 precipitates with antiesterase-2 from the rabbit. 6. It differs from esterase-2 by its individual substrate preference and by its susceptibility to inhibition by acylating agents.


Comparative activities of glycogen phosphorylase and gamma-amylase in livers of carp (Cyprinus carpio) and goldfish (Carassius auratus). 1. The activity of glycogen phosphorylase in goldfish liver is fivefold greater than that in carp liver, suggesting that the enzyme may not be as important in regulating glycogenolysis in the latter species. 2. The activity of gamma-amylase is comparable in carp and goldfish liver. 3. The activity of hepatic gamma-amylase is approximately one-half that of glycogen phosphorylase in carp whereas in goldfish, the activity of gamma-amylase is less than one-sixth that of phosphorylase. Hepatic gamma-amylase may be an important glycogenolytic enzyme in carp but makes an insignificant contribution to glycogenolysis in goldfish.


Comparative properties of mammalian and insect carbonic anhydrases: effects of potassium and chloride on the rate of carbon dioxide hydration. 1. Carbonic anhydrase (E.C.4.2.1.1) catalysed CO2 hydration was studied with enzymes from mammalian and insect sources at CO2 concentrations of 7.6-30.8 mM. 2. At 0.01-0.15 M, potassium chloride (KCl) or choline chloride (ChCl) markedly inhibited all 8 mammalian enzymes studied. 3. Inhibition by KCl is always greater than that associated with ChCl. 4. KCl non-competitively inhibits and choline chloride competitively inhibits bovine carbonic anhydrase. 5. Carbonic anhydrase obtained from fat body, integumentary epithelium and midgut tissues of larval tobacco hornworms, Manduca sexta, is greatly stimulated by KCl and slightly inhibited by ChCl. 6. We propose that the effect of K+ on mammalian and insect carbonic anhydrases if fundamentally different.


Growth of school children with polychlorinated biphenyl poisoning or yusho. In 1968, a mass poisoning due to polychlorinated biphenyls or yusho occurred in Japan. It was reported that poor growth in school children with yusho was observed 1 year after the poisoning. This study was intended to ascertain whether or not the unfavorable effect of the polychlorinated biphenyl poisoning on growth among the school children with yusho is still appreciable. Data from the physical examinations made in 1970 and 1971 were collected and analyzed. The result obtained was that growth in yusho cases was suppressed temporarily and thereafter increments tended to be close to the average value in the control group.


Thyrotropin-releasing hormone and amphetamine: a comparison of pharmacological profiles in animals. 1. The pharmacological effects of TRH are compared to those produced by d-amphetamine in an attempt to elucidate the mechanisms underlying the activity of this endogenous peptide. 2. Although numerous amphetamine-like actions have been attributed to TRH, several differences have been noted between these compounds and are discussed. 3. At present, it is impossible to propose a single mechanism of action to explain the behavioral effects of TRH.


Effects of apomorphine and sodium Di-n-propylacetate on the aggressive behaviour of three strains of mice. 1. The effects of apomorphine and sodium Di-n-propylacetate (DPA, sodium valproate) on pain-induced aggressive behavior were investigated in three inbred strains of mice: BALB/c, C57B1/6 and DBA/2, which exhibited spontaneously low levels of aggression. 2. Apomorphine elicited aggressive behavior in the three strains, the range of effective doses being different for each strain of mice. 3. Di-n-propylacetate was effective in inhibiting apomorphine elicited aggression but the three strains exhibited a different sensitivity to this drug. 4. The effects of Di-n-propylacetate were not related to pain sensitivity, posture and locomotion. Only C57 strain exhibited a slight postural and locomotor impairment when injected with a higher dose of Di-n-propylacetate. 5. The results are discussed in terms of a genetic inference and of biological differences existing between these three strains.


Transplantation of pancreatic islet tissue and the control of diabetes mellitus. The major advances achieved in recent years in pancreatic islet cell transplantation in animals and man are presented and reviewed. In animals, the biochemical and structural changes present in experimentally induced diabetes mellitus can be reversed in part or whole following transplantation. Fetal or neonatal pancreatic islet cell preparations are probably the most appropriate tissue types to employ in the human situation. The optimal conditions for their maintenance to allow such an approach to the therapy of diabetes are under active investigation.


The effect of luteinizing hormone releasing hormone (LHRH) antiserum administration on gonadotropin secretion in the rhesus monkey. Single iv injections of a rabbit antiserum to synthetic LHRH promptly suppressed serum LH and FSH concentrations in ovariectomized rhesus monkeys. Gonadotropin levels remained depressed for 10-21 days, the approximate duration of enhanced LHRH binding activity in the circulation. Doses of LHRH antiserum sufficient to reduce tonic gonadotropin secretion did not modify the time course or magnitude of estrogen-induced gonadotropin surges. These negative findings, however, cannot be interpreted to signify that such surges are not caused by a release of LHRH.


Perfusion of rat testes and accessory sex organs: a new method. Rat testes and accessory sex organs were perfused in situ by recirculating an artificial medium through the hemicorpus preparation previously developed for studies of skeletal muscle. The advantages and limitations of this system for studying the male productive tract were examined. The electrolyte and gas composition of the perfusate remained constant and glucose levels did not fall below normal during 3 h of perfusion. Testicular water content, temperature, and ATP and GTP levels were normal at 90 min. The mean arterial pressure was 40 mm Hg and the flow rates, measured with microspheres, were normal to high to the caput epididymides, ventral prostate and seminal vesicles and approximately half normal to the testes in preparations from 90 day old rats perfused at 35 ml/min. Administration of vasodilators indicated the absence of significant vasoconstriction in the hemicorpus. There was appreciable testosterone metabolism by the preparation and in addition, there was absorption of testosterone by the plastic tubing of the perfusion apparatus. Testosterone levels in the perfusate rose for 90 min in response to hCG. There was a dose-response relationship between hCG (20-1000 mIU/ml of medium) and testosterone levels at 90 min. FSH, prolactin, insulin and vitamins had no significant effect on hCG-stimulated testosterone levels. This perfusion system should prove useful for studies of hormone action.


Effect of progesterone and its 5 alpha-reduced metabolites on gonadotropin levels in estrogen-primed ovariectomized rats. In an effort to determine whether the metabolic conversion of progestrone may be important in the feedback effects of this steroid, serum LH and FSH levels were measured after administration of progesterone, 5 alpha-dihydroprogesterone or 3 alpha-hydroxy-5 alpha-pregnan-20-one to estrogen-primed ovariectomized rats. A single injection of 2 or 4 mg progesterone, 4 mg 5 alpha-dihydroprogesterone, or 4 mg 3 alpha-hydroxy-5 alpha-pregnan-20-one 72 h (Day 3) after estrogen pretreatment induced a highly significant increase in serum LH and FSH 6 h later (1800 h). Although serum gonadotropin levels had begun to decrease 12 h after administration of the progestins, they were still significantly higher than control values and did not return to baseline levels until noon on Day 4. When either progesterone or 3 alpha-hydroxy-5 alpha-pregnan-20-one was administered at noon on Days 3 and 4, there was a significant reduction in LH levels 6 h after the second injection. In contrast, serum LH levels were slightly elevated 3 to 6 h (1500 to 1800 h) after the second injection of 5 alpha-dihydroprogesterone and did not decrease until 2100 h. There was no effect on FSH concentrations after a second injection of any of the progestins. Loss of uterine luminal fluid was observed within 24 h after a single injection of progesterone. Neither of the 5 alpha-reduced metabolites had an effect on uterine ballooning until after the second injection, and, even then, nonfluid-filled uteri were observed in only 20 to 30% of the animals. The results suggest that the conversion of progesterone to 5 alpha-dihydroprogesterone and 3 alpha-hydroxy-5 alpha-pregnan-20-one by neuroendocrine tissues may be necessary for the positive and negative feedback effects of progesterone on gonadotropin secretion. Thus, the diverse effects of progesterone may be due to progesterone per se (e.g., in the uterus) and/or its metabolites (e.g., in the hypothalamus and pituitary).


Stimulation of thyroxine-binding globulin synthesis by isolated rhesus monkey hepatocytes after in vivo beta-estradiol administration. The rate of in vitro production of thyroxine-binding globulin (TBG) was studied in hepatocytes isolated from 6 control rhesus monkeys (serum TBG: 19.6 +/- 0.5 micrograms/ml; mean +/- SE) and 6 monkeys treated for 4-5 weeks with beta-estradiol (E2) (serum TBG: 45.1 +/- 1.8 micrograms/ml). Incorporation of [3H]leucine into intracellular soluble and particle-bound TBG, and into secreted TBG was determined for incubation periods up to 9 h. TBG was purified by affinity chromatography and measured by specific immunoprecipitation. The absolute amount of [3H]TBG and the ratio of [3H]TBG to total labeled protein in the same fraction were 3-fold higher in the particulate fraction and in the incubation medium of hepatocytes isolated from E2-treated monkeys. In separate experiments, TBG accumulation in the medium was measured for periods up to 19 h by radioimmunoassay. A 2.4-fold increase was observed with hepatocytes from E2-treated monkeys (3.48 ng TBG/h/10(7) cells, compared to 1.46 in controls). Correction of the production rates for the number of cells surviving during the incubation, and assuming 10.2 x 10(9) cells per liver, gave TBG production rates of 250 micrograms/liver/day in hepatocytes from E2-treated monkeys and 104 micrograms/day in hepatocytes from control monkeys. These experiments demonstrate that estrogen increases in vitro synthesis and secretion of TBG by isolated hepatocytes. The observed 2.4 to 3-fold increase was similar to the 2.9-fold increase in TBG production measured in vivo by kinetic analysis of TBG metabolism.


Release of luteinizing hormone releasing hormone and thyrotropin releasing hormone from a synaptosome-enriched fraction of hypothalamic homogenates. A mitochondrial fraction prepared from homogenates of rat hypothalamic tissue was found by means of electron microscopy to be enriched with synaptosomes. The release of luteinizing hormone releasing hormone (LHRH) and thyrotropin releasing hormone (TRH) from this preparation was investigated. After incubation, the synaptosomes were re-isolated by ultrafiltration; and the concentration of LHRH and TRH in the ultrafiltrate was determined by radioimmunoassay. When the synaptosome-enriched preparation was incubated in 0.32M sucrose at 1 or 30 C, less than 10% of the total LHRH and TRH was recovered in the ultrafiltrate. The two hormones were released by depolarizing concentrations (60 mM) of K+ in a Ca++-dependent manner, and the stimulatory effect of K+ was essentially complete within 2 min. In the presence of 2 mM Ca++, the release of LHRH and TRH increased with increasing K+ concentrations in the range 30-120 mM. Prostaglandin E2 (PGE2), PGF2 alpha, and PGF2 beta had little if any effect on LHRH or TRH release. When the synaptosome-enriched fraction was incubated in Hanks' balanced salt solution, the release of LHRH and TRH was about 10 times greater than that seen in 0.32M sucrose. It is concluded that a synaptosome-enriched fraction from the hypothalamus contains readily releasable pools of LHRH and TRH which are mobilized rapidly by depolarizing concentrations of K+ in a Ca++-dependent manner.


Computed tomography of the spinal canal and cord. Selective cases have been presented to demonstrate the usefulness of computed tomography in disorders of the spine and spinal cord. The osseous structures are more readily demonstrated than the soft tissues. The cord is difficult and inconsistently identified. However, cystic lesions (syringomyelia) may, at times, be demonstrated. The transverse projection of the CT scan is an added benefit in demonstrating bony spinal lesions. The use of metrizamide with faster scanners should improve the usefulness of this technique in evaluating intraspinal lesions.


Cholecystokinin and secretin prevent the intestinal mucosal hypoplasia of total parenteral nutrition in the dog. Because the pancreas undergoes involutional changes during total parenteral nutrition (TPN) and because pancreatico-biliary secretions are trophic to the intestine, we studied jejunal and ileal structure and function and exocrine pancreatic function before and after 6 weeks of TPN in two groups of beagle dogs, one of which had TPN alone, the other having TPN plus daily stimulation of pancreatico-biliary secretions with intravenous infusions of cholecystokinin (CCK) and secretin. The injections of 1 U each per kg of body weight per day of CCK and secretin completely prevented the proximal and distal small bowel mucosal hypoplasia which developed in the TPN alone group. They also resulted in significant increases in in vivo galactose absorption (64 mM) per unit length of jejunum and ileum. However, there was no significant change in mucosal alpha-glucosidase and catalase activity or in in vitro mucosal uptake of 1 mM [14C]leucine when expressed per unit weight of intestinal mucosa. The capacity of the pancreas to respond to CCK and secretin was unaffected by excluding food from the intestine with 6 weeks of TPN in terms of pH, volume, and peak secretion rates of bicarbonate and protein, but maximum amylase output (units per 15 min per kg of body weight) fell significantly (P less than 0.05) from a mean of 1022 +/- 155 to 874 +/- 426 in TPN alone group and to 472 +/- 79 in the TPN dogs given CCK and secretin. These results show that daily CCK and secretin is trophic to the intestine of dogs nourished by TPN but do not indicate whether this trophic effect is attributable to CCK alone, secretin alone, the combination of the two hormones, or to the resultant stimulation of pancreatico-biliary secretions.


Thyrotropin-releasing hormone (TRH)-induced growth hormone (hGH) responses in cirrhotic men. The plasma growth hormone (hGH) responses to an intravenous challenge of 400 micrograms of thyrotropin-releasing hormone (TRH) were evaluated in 14 normal controls and in 29 chronic alcoholic men. The normal controls had either a minimal or no hGH response to TRH, having basal hGH levels of 0.9 +/- 0.2 ng per ml and peak hGH levels of 2.0 +/- 0.5 ng per ml. In contrast, the chronic alcoholic men had a basal hGH level of 2.8 +/- 0.4 ng per ml, 3 times the basal level of the normal controls (P less than 0.01). The peak hGH response of the alcoholic men was 7.4 +/- 1.5 ng per ml (P less than 0.01). The 29 alcoholic men could be divided into two groups based upon the presence or absence of cirrhosis as determined by liver biopsy. The 16 alcoholic men with cirrhosis had greater basal hGH levels (3.5 +/- 0.6 ng per ml) and peak hGH levels (9.5 +/- 2.3 ng per ml) than did the 13 alcoholic men without cirrhosis (basal hGH 2.1 +/- 0.6 ng per ml, peak hGH 4.9 +/- 1.5 ng/ml). Plasma estradiol levels were similar in the normal controls and in the alcoholic men. In contrast, plasma estrone was greater in the alcoholic men (32.2 +/- 3.5 pg per ml) than in the normal controls (18.9 +/- 1.8 pg per ml) (P less than 0.05). However, when the plasma estrone levels of alcoholic men with cirrhosis were compared to those of the alcoholic men without cirrhosis no difference existed. Thus it is difficult to ascribe the increased hGH responses of the cirrhotic alcoholic men when compared to those of the noncirrhotic alcoholic men as being a result of increased basal estrogen levels.


Neonatally thymectomized Lewis rats infected with Mycobacterium leprae. 2. Histopathologic and electron microscopic observations. We report the histologic and electron microscopic findings following intravenous inoculation of M. leprae into neonatally thymectomized Lewis rats, which were killed one to two years later. All organs appeared normal grossly. Histologic changes were confined to the footpads, snout, ears, tail, and testes, all of which were involved in every rat. The tissues were edematous and infiltrated by varying numbers of foamy macrophages. In the footpads muscle fibers were vacuolated, and small nerves showed degenerative changes. Large numbers of M. leprae were present in macrophages and striated muscle cells and smaller numbers in perineural cells and pericytes, as well as lying free in the tissues. Occasional intracellular bacilli were found throughout the reticuloendothelial system. Electron microscopy confirmed that the majority of organisms were within activated macrophages. Both intact and fragmented bacilli were contained within double-membrane bound vacuoles. Numerous M. leprae were lying free within the sarcoplasm of striated muscle cells. Virtually all of the extracellular organisms were degenerating.


In vitro activation of neutrophils by suspensions of Mycobacterium leprae. Activation, defined as an increase in the proportion of cells that reduce nitroblue-tetrazolium in vitro, is present in neutrophils from patients with reactional lepromatous leprosy but not in neutrophils from patients with non-reactional lepromatous leprosy. Neutrophils from patients with all forms of leprosy are equally well activated by endotoxin in vitro. We have now shown that in vitro activation induced by Mycobacterium leprae suspensions is of comparable magnitude in neutrophils from patients with all forms of leprosy (including lepromatous and reactional lepromatous leprosy). There is no intrinsic neutrophil anergy in patients with lepromatous leprosy vis-à-vis M. leprae as pertains to activation. Spontaneous activation in reactional lepromatous leprosy is likely due to an indirect mechanism, probably of immunologic nature, and not simply to the presence of circulating Mycobacterium leprae in the blood.


Defective blood mononuclear phagocyte function in patients with leprosy. Patients with lepromatous leprosy possess a defective lymphocyte function in vivo and in vitro that is less evident in the tuberculoid form. Data concerning their macrophage ability to digest Mycobacterium leprae are controversial. The purpose of this study was to determine whether monocytes from patients with either tuberculoid or lepromatous leprosy were altered in their enzyme systems, that is myeloperoxidase-dependent and myeloperoxidase-independent systems. The ability of adherent blood monocytes to ingest and kill Candida pseudotropicalis after 30 and 60 min of incubation with yeast cells was tested. Mononuclear phagocytic cells from patients with either principal form of leprosy functioned similarly to normal monocytes in phagocytosis while their fungicidal activity for C. pseudotropicalis was statistically significantly altered and was more evident in the lepromatous than in the tuberculoid type. The results indicate that peripheral blood monocytes from patients with leprosy possess an impaired enzymatic candidacidal activity.


Leprosy and hepatitis B virus markers: incidence of HBsAg and HBeAg in Somalian patients. Serum samples from 222 Somalian patients, 135 with the lepromatous form of leprosy and 87 with the tuberculoid form of the disease, were examined for the presence of the surface antigen (HBsAg), the "e" antigen (HBeAg), and their corresponding antibodies (anti-HBs and anti-e). HBsAg was present in 24.4% of the LL cases and in 11.5% of the TT patients while anti-HBs was found respectively in 46.6% and 58.6%. The e-antigen was not found in any case of leprosy; anti-e was detected in 8.1% of the LL patients and in 3.5% of the TT cases. The rate of HBV seropositivity (HBsAg plus anti-HBs) was the same in the LL patients (71.1%) and in the TT patients (70.1%) and that could reflect the conditions of life in their closed community. The analysis of results obtained in Somalia has shown the presence of a difference in the distribution of HBsAg among leprosy patients, with an increased antigenemia in the lepromatous form which was statistically significant (p less than 0.05). No differences, however, were found between the leprosy patients and healthy controls. These observations seem to indicate that patients with lepromatous leprosy do not have an increased susceptibility to infection by hepatitis B virus.


Electron microscopic observations of intracytoplasmic membrane systems and cell division in Mycobacterium lepraemurium. The fine structures and the interconnections between the intracytoplasmic membrane systems and cell division of murine leprosy bacilli in liver tissue from mice infected intraperitoneally with Mycobacterium lepraemurium, Hawaiian strain, were studied in ultrathin serial sections at the electron microscopic level. Intracellular membranous organelles (mesosomes) were seen as vesicular, tubular and/or lamellar structures. The formation of mesosomes appeared to be initiated by invagination and/or folding of the cytoplasmic membrane. A few dividing bacilli were observed. The lateral extension and the centripetal growth of the septal wall seemed to result from the original development of cytoplasmic membrane occurring at or near the leading edge of the nascent septum. After the septum formation was completely accomplished, the separation of two new daughter cells is assumed to occur. The mesosome was associated with the newly formed cytoplasmic membrane (septal wall). It was shown that both the cytoplasmic membrane and the mesosome played an important part in septum formation.


Histocompatibility antigens in patients with leprosy from Azarbaijan, Iran. The distribution of 24 histocompatibility antigens in 88 Azerbaijani patients with leprosy was determined and compared with those of 125 normal, ethnically matched individuals. HLA-BW35 was increased in frequency among the Kurdish patients as compared to the controls; HLA-A1, however, displayed decreased frequency in patients with the lepromatous form of the disease. Among the Turks, diminished frequency of HLA-BW15 was noted in the total patient population. None of these comparisons, however, reached statistical significance when corrected for the number of antigens tested. Across the two ethnic groups, differences in the frequencies of HLA antigens between the patients and the controls were only marginal.


The potential uses of the medication monitor in the treatment of leprosy. A medication monitor has been developed that utilizes radioactive material and photographic film to record the intervals at which patients take medication. In the author's opinion, this equipment represents the most efficient means that has so far become available for determining how regularly outpatients take medication. A monitor for the tuberculosis regimen of isoniazid and thiacetazone has been made at a sufficiently low cost that it is practical to use it in routine treatment programs. An inexpensive system for immediate development of the film that can be used in the most remote locations is also available. Undoubtedly, with appropriate engineering work, a monitor for leprosy regimens could be made. The device has been used with tuberculosis patients and revealed that many patients were grossly irregular in taking their medication. It has been used to oversee medication use by tuberculosis patients and to select those who require either extra attention to improve medication ingestion or completely supervised, directly administered programs. In the treatment of leprosy, it could be used to study new drug regimens, the causes of noncompliance, and for the routine supervision of patients.


Miconazole in coccidioidomycosis.

Miconazole in coccidioidomycosis. The epidemiology of Coccidioides immitis is described with respect to annual infection rates and differing ethnic susceptibilities to the infection. A trial of miconazole in infected mice produced 100% survival in conditions which normally kill 100% of animals, probably by drastically reducing the rate of fungal multiplication in the lung. The fungal endospore, the form of the dimorphic fungus found predominantly in host tissue is more sensitive to miconazole than is the mycelial form. Data are presented from 14 human patients, some moribund and comatose, all of whom had failed to respond to existing therapy. There was objective evidence of improvement in eight, and a subjective impression of improvement in the majority. One patient failed to respond and three have equivocal results.


Miconazole in systemic candidiasis.

Miconazole in systemic candidiasis. Based on our experience with our first patients, miconazole is now the drug of choice in cases of systemic and pulmonary candidal infection. Initially we prefer the parenteral route of administration which is continued for only one or two weeks after a negative finding in culture. The dosage is 1.2 g per day for a 70 kg patient. Thereafter the treatment is continued orally for another four weeks. Miconazole is well documented as a drug without serious side-effects, but the incidence of candidal infections in about 1% of our surgical patients does not justify its prophylactic administration. Like antibiotics, miconazole is given only when there is a clinical manifestation and a positive finding in culture.


Therapy for deep mycoses: an introduction. A classification of mycoses, depending upon the depth of penetration of the organism is discussed, as is the need to distinguish the true from the 'pseudo' mycoses. The development of antifungal agents is described from the first antibiotics to amphotericin B and 5-fluorocytosine. Attention is particularly focussed on the question of safety and effectiveness. Finally, it is stressed that successful antifungal treatment must depend upon determination of the exact pathological status of the patient and examples are given which relate to such clinical assessments.


Miconazole in systemic candidosis.

Miconazole in systemic candidosis. Results are reported on the effect of miconazole together with concurrent medication in the treatment of 7 patients with systemic candidiasis. Two patients were discharged from hospital in good condition. In two others, the clinical status normalized. One patient's status improved but he was subsequently readmitted. Two patients died. The excellent tolerance of miconazole by all patients was a particular feature of this trial.


Results of miconazole therapy in twenty-eight patients with paracoccidioidomycosis (South American blastomycosis). Results are presented of treatment with miconazole, orally and intravenously, in patients with paracoccidioidomycosis. Twenty-eight male patients aged from 34 to 66 years and exhibiting various clinical forms of the disease were studied. Twenty-five came from endemic areas in north east Argentina (Chaco, Formosa, Misiones, Corrientes and northern Santa Fe) and the remaining three from Paraguay. Twenty patients were engaged in agricultural work or at woodmills. single or multiple lesions were observed in 24 cases. Thirteen were suffering from infection of the larynx and in two of them a tracheotomy was necessary. Twenty-three showed pulmonary lesions on X-rays. Twelve had ganglionic lesions, eight had cutaneous lesions and one patient had osteoarthritis of the knee. One patient had hepatomegaly which was unrelated to chronic alcoholism. Fourteen patients had received previous treatments such as sulphonamides and amphotericin B (7 cases); sulphonamides (3), sulphonamides and the combination sulfamethoxazole + trimethoprim (3), and one patient had received all three medications. All patients had relapsed before starting miconazole therapy. Diagnosis was established by the presence of P. brasiliensis in all cases, recovered either from cutaneous or mucosal biopsy samples or from the sputum. Complement fixation tests were positive in all patients at the onset of the treatment and the immunodiffusion reactions showed precipitation bands in 27/28 patients. Skin tests with P. brasiliensis antigens proved to be positive in 18 cases and negative in 10. The erythrocyte sedimentation rate was markedly accelerated in 22 patients (greater than 20 mm in the first hour).(ABSTRACT TRUNCATED AT 250 WORDS)


Miconazole in oral candidiasis.

Miconazole in oral candidiasis. Twenty-four patients were treated with oral miconazole (250 mg) for a total of 35 episodes of oral candidiasis. Sixteen had various forms of leukaemia and all were massively predisposed to fungal infection because of granulocytopenia and treatment with prednisolone and antibiotics. Clinical cure was observed in all 35 of the treated episodes, with a mean treatment time of five days, cure being observed in two to three days. When patients violating the protocol were excluded, the mycological cure rate was 97%. In 21 episodes there was a recurrence less than one month after miconazole treatment, probably because of reinfection. No side-effects ascribable to miconazole were observed, even in the severely debilitated patients, and the orally administered drug appeared to be superior to other commercially available antimycotic preparations.


Use of miconazole for prevention of opportunistic fungal infection during treatment of haematological malignancies. It appears that miconazole is highly effective in reducing the incidence of systemic mycosis in patients suffering from malignant haemopathy and bone marrow failure. Three clinical infections (one stomatitis and two septicaemias due to candida) were easily cured. Autopsy findings never disclosed mycosis as the cause of death. The drug was completely atoxic and seems to offer major help in the treatment of malignant blood disease.


Treatment of South American blastomycosis (paracoccidioidomycosis) with miconazole by the oral route: an on-going study. Results are presented from 12 patients of both sexes, whose ages range from 34 to 65 years, observed over a period of 16 months and receiving miconazole treatment for South-American blastomycosis. Ten of them were being treated for the first time and the remaining 2 presented relapses after treatment with other chemotherapeutic agents. The blastomycotic lesions were localized in skin, mucosa, lung and lymphatics. The drug was administered orally, at a dosage of 1 g three times daily. One relapsing patient had his first daily oral dose replaced by an intravenous dose of 200 mg of miconazole diluted in 250 ml of glucose solution during the initial four weeks. The criterion for diagnosis was finding of the fungus in sputum and/or in the material collected through scrapings or biopsy of the lesions. All patients were treated in hospital until the cure was apparent, clinical examinations being conducted weekly and bi-weekly. Bi-weekly laboratory examinations were performed to evaluate the patient's tolerance to the drug. All treated patients presented complete healing of cutaneomucous lesions (mean time: 4 weeks) and involution of lymphatic and pulmonary features (mean time: 4 months). Two of them still presented palpable lymphatics after 5 months of treatment, in spite of the cure of mucosal and pulmonary lesions. Monitoring the progress of the cured patients was and is being carried out quarterly. Currently, this observation period ranges from three to 15 months. Nine patients are still returning and none have shown the reappearance of mycotic lesions. Diarrhoea was the only side-effect observed after a period of therapy, but it was easily controlled by specific medication (kaolin, pectin, &c.). In a single case the treatment was interrupted because of this symptom.(ABSTRACT TRUNCATED AT 250 WORDS)


Clinical and experimental evidence on miconazole for the treatment of systemic mycoses: a review. Intravenous treatment with miconazole brought about the recovery of 90% of patients with gastrointestinal or systemic candidosis. Miconazole given by the same route has also been found effective in the treatment of cryptococcosis, coccidioidomycosis, and paracoccidioidomycosis. Cryptococcal and coccidioidal meningitis have been cured by combined intravenous and intrathecal instillation, although treatment of aspergillosis has presented difficulty. Oral treatment was effective in curing dermatophyte skin infections and systemic mycoses caused by sensitive organisms such as paracoccidioides, blastomyces and histoplasma. The question of blood levels following oral and intravenous administration is discussed. Side effects of the drug were few, and included chills, dizziness, skin rash, itching and diarrhoea. Thus miconazole can safely be given to seriously ill patients. Its behaviour in the body is not influenced by renal insufficiency and no drug induced resistance has been reported.


Treatment of fungal infections with miconazole. The present study reports on 15 patients with confirmed or suspected secondary fungal infection, who were treated with intravenous miconazole and a daily dose of 0.4 to 28 g (mean 9.8 g). The mycological findings confirmed infection with Aspergillus fumigatus in 6 patients and severe infection with Candida albicans in 4. Clinically mycological cures were achieved in 4 patients and some improvement was found in 3 others. The excellent patient tolerance of the drug makes it seem a likely treatment not only for patients with mycologically confirmed infection but also for those whose condition exposes them to a high risk of fungal infection.


Miconazole treatment in candidal oesophagitis. Seven patients with candidal oesophagitis were treated for up to 8 weeks with an intravenous infusion of miconazole, usually 600 mg given over a period of 30-60 minutes with a frequency depending upon the individual patient. As well as oesophagoscopy, biopsy and culture to establish the clinical course of the disease, a series of immunological investigations were performed, to assess the involvement of the patients' immune mechanisms, together with a whole battery of laboratory investigations to assess their response to the drug. A subjective improvement was observed in all patients after periods varying from 3 to 10 days and objective improvements were observed by oesophagoscopy by 5 of the patients in periods of one to four weeks. The same procedure showed 3 patients to have become completely normal after periods of one to three months. Apart from one instance of nausea, no other side-effects and no abnormal laboratory results were seen. It is suggested that the results of treatment depend on the clinical status of the patient, the associated pathology and the underlying immunological disturbance.


Effect of penicillamine on the kidney.

Effect of penicillamine on the kidney. Twenty patients with rheumatoid arthritis developing proteinuria when being treated with penicillamine have been studied. In 5 the proteinuria was mild and resolved rapidly. Eleven of the remaining patients have undergone renal biopsy. One was found to have amyloidosis, and in the other 10 there was evidence of an immune complex type of injury, manifested by a granular immunoglobulin deposition within glomerular capillary walls associated with subepithelial deposits, found on electron microscopy. The proteinuria associated with this was mild (mean 2.6 g per 24 h) and occurred on average some eight months after commencing therapy (range 6 weeks to 60 months).


Penicillamine treatment of rheumatoid arthritis: relationship of proteinuria and autoantibodies to immune status. Thirty-seven patients were studied before and during treatment with respect to immune status, clinical response and development of adverse effects and autoantibodies. The baseline immune status was not predictive in terms of the above features, apart from the fact that the group of 7 patients developing proteinuria had a tendency to low or subnormal IgG levels. The most marked clinical improvement was recorded in the group who had augmented skin test responses sometime during the treatment period. These patients also had the largest falls in the IgG, IgA and rheumatoid factor. Antinuclear antibody persisted or increased in titre in 38% of patients, but was not associated with poor prognosis or liability to side-effects. Autoantibodies to striational or smooth muscle occurred in 20% of patients, and there was a much higher incidence of proteinuria in this group. We have previously suggested that penicillamine may act by depressing humoral function, leading to augmentation of cell-mediated immunity. Although the present findings suggest that penicillamine does cause humoral depression in some cases, it is not clear how the drug induces the side-effects described.


The effect of penicillamine on growth as height in juvenile chronic polyarthritis. When the disease state improves during penicillamine therapy, resumption of growth occurs; if activity persists despite therapy, the expected fall-off in growth due to active Still's disease continues. Penicillamine does not interfere with the growth spurt after withdrawal of corticosteroid therapy or with the maintained slow growth during alternate-day steroid therapy.


Low Dose D-penicillamine in cystinuria.

Low Dose D-penicillamine in cystinuria. (1) A single dose of D-penicillamine (not more than 750 mg) taken at 2200 h, together with a prescribed fluid intake of two to three litres during the waking hours, without extra drinks at night, is sufficient to keep the concentration of the urinary cystine below the saturating concentration of urine throughout the 24 h period. (2) This regime does not materially reduce the total 24 h excretion of cystine. (3) The effect of D-penicillamine is mainly seen in the urine excreted between 0200 and 0800 h. (4) This regime is provisionally recommended for the prevention of cystine stone recurrence but not for stone dissolution. Larger total amounts of D-penicillamine given in divided doses are still recommended for the latter purpose.


An outline of D-penicillamine metabolism. In a study using 14C-D-penicillamine in a normal subject only 50% of the D-penicillamine was absorbed from the gut. The major urinary metabolites are cysteine-penicillamine and penicillamine disulphide. S-methyl-D-penicillamine is the only biochemically transformed metabolite so far identified.


Penicillamine therapy in systemic sclerosis. Twenty-two patients with progressive systemic sclerosis were treated with D-penicillamine in doses ranging up to 1250 mg/day for periods varying between a few months and four years. Side-effects occurred in 7 patients, necessitating drug withdrawal in 2. Cutaneous benefit occurred in 15 patients, but owing to side-effects from the drug, relapses, and development, persistence or advancement of visceral complications, an overall good result only occurred in 5. Seven patients showed improvements in joint function, but only 3 were regarded as having an overall good result. Peripheral vascular disease and visceral involvement seemed not to be influenced by D-penicillamine and sometimes appeared or advanced during treatment. Six patients died from visceral manifestations of systemic sclerosis and one from another cause. D-penicillamine is of limited value for the cutaneous features of progressive systemic sclerosis, but probably of no value for the vascular and visceral manifestations of the disease.


Ultrastructural study of apatite crystal dissolution in human dentine and bone. The width and thickness of normal apatite monocrystals were determined in human dentine and alveolar bone on non-decalcified thin sections made perpendicular to the c axis. A mean width (L) of 364.50 A +/- 14.50 A and 562.10 A +/- 19 A was respectively found for dentine and bone monocrystals, whereas their mean thickness (E) was respectively 103.30 A +/- 2.70 A and 79.10 A +/- 3.10 A. The study of the ratio L.E.-1 for dentine and bone monocrystals showed a plate-like configuration which was more accentuated in bone crystals. Apatite crystal dissolution was studied during the carious process in dentine and bone resorption during advanced periodontal lesions. It could be shown that, in both conditions, one or several central core lesions developed along the c axis of the crystals. These central core lesions, extending laterally along (100) planes, progressively fused together, leading to the splitting up of the monocrystals into two thin plates by a complete destruction of the central part of the monocrystal.


Minor elements in bone mineral and their effects on its solubility. Mineral substance deposited in bone tissue when either strontium or fluoride are present, is both qualitatively and quantitatively different from that deposited in their absence. The presence of these ions causes not only heteroíonic substitutions in the crystalline lattice of the apatite phase, but also important modifications of the percentage of crystalline mineral, of crystal size and perfection, and of quantity of mineral per unit volume. These modifications affect the in vitro and in vivo demineralization of bone tissue.


Some ultrastructural aspects of biological apatite dissolution and possible role of dislocations. High resolution electron microscope techniques now make it possible to study the mineralization of calcified tissues at the crystal structure level. Dislocations play an important part in the course of crystal maturation and modifications. At least two origins of dislocations are known. The first is due to phenomena related to the incorporation of fluoride ions into the lattice. The second is due to mechanical stresses occurring between crystals during their maturation. Dislocations are the starting-points of acid dissolution which proceeds along dislocation-lines, sometimes inducing a splitting of the crystals. In the present study, dislocations have been visualized, either isolated in the crystal core and perhaps the two-dimensional surface representation of a screw-shaped dislocation, or forming nets of dislocations producing a spiral staircase on the sides faces of the crystals. There is evidence of a maturation cycle of the crystals, which may grow, split, coalesce or dissolve; liberated elements and small crystal fragments may allow development of near-by crystals. This remodelling explains the existence of a standard size as well as the perfect fitting of the crystals. These morphological data involve physico-chemical properties of biological apatites.


Prevention of denture plaque formation by an enzyme denture cleanser. It was the purpose of the study to test the efficacy of dissolvent tablets containing mutanase and protease in preventing formation of plaque on the fitting surface of complete dentures. The study group consisted of 60 denture wearers with denture stomatitis who were assigned randomly into an enzyme group, a placebo group, or a Steradent group. After denture treatment was completed the patients were instructed to immerse the new dentures for 15 min once daily for one month in the denture cleanser. The amount of denture plaque, the clinical condition of the palatal mucosa, and the concentration of yeasts in mucosal and denture smears were recorded while the patients used their original dentures and after the experimental period. The study was designed and carried out as a double-blind study. New denture plaque had formed in all patients; however in the enzyme group significantly reduced plaque scores were recorded as compared with the plaque scores recorded on the original dentures. The Steradent tablets or the placebo tablets had no apparent effect.


[The role of hypocalcemia on cell renewal and migration in the oral epithelium of the rat]. Calcium plays an important role in phenomena relating to permeability and regulation of the extracellular compartments. The diffusion anomalies of sulphur previously observed in the oral epithelium of the hypocalcemic rat could possibly be attributed to a reduction in diffusion of sulphur related to disturbances in cellular renewal. The injection of H3 thymidine was used to compare in parathyroidectomized and control rats the resultant differences in DNA synthesis and the migration of labelled cells. The labelling indices showed the absence of any relation between DNA synthesis and a calcium deficiency, whereas, on the contrary, hypocalcemia should produce modifications of the labelled cell distribution in the control and experimental rats. This difference indicates a reduction in cellular migration in case of hypocalcemia.


[A technic to prepare fixed ground sections for a quantitative microradiography study of the dentin]. The preparation of ground sections for the quantitative microradiographic study of dentine was carried out either by hand or by machine using a new technique of fixing the sections to the holder by a soluble adhesive. After setting up a method of control for the plano-parallelism of the sections, the times of preparation and the quality of the sections obtained with both methods were compared: engine grinding gave the best results in terms of both quantity and quality.


[The morphologic relation between the temporomandibular joint and other components of the masticatory apparatus in catarrhine simians and humans]. The articular temporo-mandibular structures and their osseous and dental environment were studied in some catarhinians Simians (Cynomorphs and Anthropomorphs) and some representatives of the human lineage. The study of measurable and descriptive characters showed a relative stability of the temporo-mandibular joint compared to the dynamism of the other components of the masticatory system. The functional specificity of the actual human temporo-mandibular joint seemed to be tightly associated to the main processes leading toward human evolution (facial regression, increase of the cranial capacity). On the other hand, this study casts some doubts regarding the utilization of Primates for experiments in this field.


Degenerative changes in the histology of the human submandibular salivary gland occurring with age. A histological investigation was made of 96 human submandibular salivary glands, obtained at necropsy and evenly representative of sex and adult age to 95 years. Age-dependent atrophy was observed in both acinar and ductal epithelia. With increasing age structural changes were also found in fibrous and elastic tissues as well as in the walls of arteries and veins. Ischaemia due to partial occlusion of arteries may be one reason for the age-dependent atrophy of parenchyma. The deteriorative changes developing with age in arterial walls and in acinar and ductal epithelia probably both contribute to the reduced functional efficiency of salivary glands which is known to occur in senescence.


Combined micromanipulation, culture and immunofluorescent techniques for isolation of the coccal organisms comprising the "corn cob" configuration of human dental plaque. The present study describes methods to 1) selectively isolate Corn Cob Configurations (CCC) from dental plaque by micromanipulation; 2) obtain pure cultures of the coccal constituent; 3) determine by immunofluorescent procedure which organisms originated from the CCC. Using a de Fonbrune micromanipulator, CCC specimens were isolated from supragingival plaque samples. The viability of one specimen thus obtained was established by observing growth on a slide culture. One set of CCC specimens was transferred to broth and incubated aerobically immediately upon collection. Another set was transferred to prereduced transport medium and later plated on blood agar for aerobic and anaerobic culturing. A total of 10 coccal strains were thus isolated. Antisera produced in rabbits against the 10 strains were used to localize these coccal organisms on plaque smears by using the indirect fluorescent antibody technique. Of the 10 antisera tested, 2 produced against streptococcal strains consistently gave a positive immunofluorescent reaction with the coccal component of CCC in the plaque smears; the corresponding streptococci were therefore considered to be CCC forming strains in vivo.


[Cathepsin D in the connective tissue and epithelium of inflammed human gingiva]. The activity of cathepsin D has been determined, as a function of gingival inflammation, in biopsies of human gingivae from 10 patients. The determinations have been performed both in the connective tissue and epithelium after their mechanical separation in cryostat sections. A positive and statistically significant correlation was found between the cathepsin D specific activity (as a function of either dry weight or DNA) and the degree of gingiva inflammation. These results support the hypothesis of a possible participation of lysosomal enzymes in the destruction of periodontal tissues during gingivitis and periodontitis.


Collagen resorption by fibroblasts in human gingiva. In the course of the extensive tissue remodeling occurring after flap surgery and during the subsequent gingival reattachment in man, numerous intracellular vacuoles were observed in fibroblasts of the lamina propria. These vacuoles contained one or several typical collagen fibrils and had either an electron-lucent matrix or contained an electron-dense material obscuring the fibrillar outlines and cross-striations.


[Ultrastructural demonstration of the phagocytic activity of periodontal ligament fibroblasts in the rat using enzymatic tracers]. The ultrastructural study of non-decalcified periodontal ligament of rats, labeled with horseradish peroxidase used as an enzymatic tracer for the mechanism of endocytosis, shows that there exists in the fibroblast, on the one hand, an endogeneous peroxidase revealed by the preparation and characteristic of certain cellular organelles: mitochondria, free ribosomes, ribosomes of the granular endoplasmic reticulum; and on the other hand an exogeneous peroxidase which demonstrates the ability of fibroblasts to phagocytose. This exogeneous peroxidase initially marks the collagen fibrils located in the extracellular medium, then at a later time he cytoplasmic membrane during its invagination, and finally the membrane of phagosomes. The intracellular collagen situated at the interior of phagosomes is also marked by the peroxidase and demonstrates the passage of the collagen from the extracellular medium to the intracellular medium.