A comparison of the effect of benzopyrones and other drugs with anti-inflammatory properties on acid and neutral protease activity levels in various tissues after thermal injury. Generally, the benzopyrones enhanced acid protease activity levels in the oedema fluid and the extracellular compartment of the skin. This is the region where thermal injury has its greatest impact. The proteolysis induced by the drugs in this region represents a means of rapidly reducing some of the derangements which the thermal injury has caused. Levamisole also enhanced acid protease activity levels in the serum and extracellular compartment of the skin 6 hours after thermal injury, while Reparil had the same effect at 24 hours. Generally the benzopyrones had little or no effect on neutral protease levels, while levamisole and Reparil caused their depression. The later effects could possibly be attributed to serum deactivation or to inhibition of their release. The enzyme enhancing activity of these drugs has been shown to correlate remarkably well with their oedema reducing ability. Generally, those which increased enzyme activity levels the most were the most effective in reducing the oedema. The cells upon which the drugs exert their effects in thermal oedema mainly seem to be the macrophages; the fibroblasts seem to be of secondary importance. This is to be contrasted with their action in the initial stages of lymphoedema where they are believed to stimulate the neutrophils. The net result of the proteolysis is many small fragments which can rapidly leave the injured tissue thus releasing the oedema fluid.


Altered nuclear deoxyribonucleic acid alpha-polymerases in senescent cultured chick embryo fibroblasts. DNA alpha-polymerase has been partially purified from nuclei of cultured chic, fibroblasts and separated on phosphocellulose columns into two distinct activities designated DNA polymerases alpha(a) and alpha(b), respectively. The enzyme preparations were devoid of activities of DNA beta,gamma-polymerases terminal deoxyribonucleoside transferase, DNase, DNA-dependent RNA polymerase, and phosphatase. DNA polymerases alpha(a) and alpha(b) both having molecular weights of 160 000, constitute 35-50 and 65-50%, respectively, of the activity of alpha-polymerase in the nucleus. These enzymes differ in their requirements for maximal activity, their relative ability to copy oligo(dG)-poly(dC), their response to ribonucleoside triphosphates, and their kinetics of heat inactivation. When the properties of alpha polymerases derived from early or late passage cultures have been compared, no difference could be detected as a function of cell age in the specific activities of the polymerases in crude cell extracts, their chromatographic behavior on diethylaminoethylcellulose and phosphocellulose columns, and their relative abilities to utilize single deoxyribonucleoside triphosphates with activated DNA template. On the other hand, both enzymes become partially heat labile in aging cells. Also, the activity of DNA polymerase alpha(a) from young cells was stimulated by 2--10 mM adenosine or cytidine triphosphates, whereas the same enzyme from old cultures was inhibited by these agents. Conversely, these ribonucleoside triphosphates inhibited the activity of polymerase alpha(b) in young cells but slightly stimulated this enzyme derived from senescent fibroblasts. In addition, the relative ability of DNA polymerase alpha(a) to copy oligo(dG)-poly(dC) decreased in aged cells, whereas that of DNA polymerase alpha(b) increased. We have also observed significant differences in the effects of potassium chloride and N-ethylmaleimide on the activity of DNA polymerase alpha(a) from old cells as compared to young cells. These age-related alterations in the properties of the two avian DNA polymerases may reflect structural or conformational changes in these enzymes.


gamma-Carboxyglutamic acid distribution. The distribution of the vitamin K-dependent amino acid, gamma-carboxyglutamic acid was examined in proteins from a variety of sources. Proteins examined include purified rat and bovine coagulation proteins, barium citrate-adsorbing proteins from trout plasma, lamprey plasma, earthworm hemolymph, army worm hemolymph, lobster hemolymph, E. coli B/5, soybean leaf, the protein lysate from the hemolymph cell of the horseshoe crab and parathyroid extract. Other purified proteins examined included human alpha-1-antitrypsin, pepsinogen, S-100, fetuin, tropomyosin-troponin and complement protein C-3. Of these, only the blood-cotting proteins and the vertebrate plasma samples were shown to contain gamma-carboxyglutamic acid.


Mammalian transcobalamin II metabolism. The immunological and the biological cross-reactivity of mammalain transcobalamin II. Assay of the reactivity between the chicken anti-rabbit transcobalamin II antiserum and the sera of 19 vertebrate species was carried out by both immuno-diffusion and Sephadex G-200 gel filtration column chromatography. Mammalian transcobalamin II cross-reacted with the antiserum whereas the serum vitamin B-12 binders of the bird, amphibian reptile and fish did not. The biological activity of the purified rabbit transcobalamin II was assessed using reticulocytes or erythrocytes of human, rabbit, guinea pig and rat. The purified rabbit transcobalamin II promoted the uptake of vitamin B-12 by the cells but showed a great variation in its activity. It is suggested that the rabbit transcobalamin II is immunologically and biologically similar to the serum transcobalamin II of the mammalian species studied.


Receptor distribution and the mechanism of enhanced erythrocyte agglutination by soybean agglutinin. We have examined the role of receptor clustering in intact erythrocyte membranes exhibiting enhanced lectin-mediated cell agglutination by analyzing freeze-fracture and freeze-etch images of human erythrocytes labeled with ferritin-conjugated soybean agglutinin. We find that trypsinization and fixation of intact erythrocytes, in either order, causes no alteration of the random distribution of ferritin-conjugated soybean agglutinin on the surfaces of these cells as compared to their distribution on the surfaces of fixed erythrocytes and untreated erythrocyte ghosts. Furthermore, clustering of the intramembranous particles in the membrane of intact erythrocytes was not found with any of the cells described above. We conclude that clustering of the soybean agglutinin receptors is not a major factor involved in the enhanced agglutination of intact trypsinized erythrocytes. Caution is necessary in transferring information obtained with erythrocyte ghosts, where clustering can be induced, to intact erythrocytes.


A new assay for endocytosis in erythrocyte ghosts based on loss of acetylcholinesterase activity. A new method for assaying endocytosis in erythrocyte ghosts is presented. The method involves measuring the percentage loss of acetylcholinesterase activity which occurs when vacuoles form, making the acetylcholinesterase on the vacuole surface inaccessible. This method is compared to other methods of measuring endocytosis in this system, including phase contrast microscope estimation of vesiculation, stereological analysis of electron micrographs to determine vesiculation and loss of sialic acid accessible to neuraminidase due to endocytosis. Comparison of the percentage loss of acetylcholinesterase activity with the electron micrographic and sialic acid methods showed that all three methods gave a quantitative measure of the percentage of total membrane area taken in as vesicles. Since the acetylcholinesterase method was fast, easy, inexpensive, and quantitative, it was the preferred method for assay of endocytosis. The inhibition of endocytosis by Ca2+ was observed with this method; the success of this experiment demonstrated the applicability of the method to the study of inhibitors of endocytosis.


Small amount of concanavalin A modifies radiation-induced alteration in cell-surface charge depending on its binding condition. Cell electrophoretic mobility of cultured melanoma cells or rat erythrocytes decreased with time after X-irradiation. Addition of tetravalent concanavalin A or divalent succinyl-concanavalin A before (not after) irradiation, completely blocked the mobility reduction in greater concentrations than 5 mug/l. At 5 mug/l only 3.7 - 10(3) concanavalin A molecules bound to receptors per cell, while 4.18 - 10(7) molecules/cell bound at saturating concentrations. Preincubation with concanavalin A at 37 degrees C was effective even when the cells were treated with alpha-methylmannoside immediately after irradiation. At low temperature, however, concanavalin A was not effective despite a sufficient amount of bound 125I-labelled concanavalin A. Treatment with alpha-methylmannoside following the binding of concanavalin A at 37 degrees C before irradiation inhibited the concanavalin A effect depending on temperature. The residual amount of bound lectin could not account for the temperature dependence. The amount of sialic acid (the main charged substance) was not altered by X-irradiation with or without the lectin. Divalent succinyl-concanavalin A was also effective in blocking the radiation effect on electrophoretic mobility. These results seem to suggest that binding of a very small amount of concanavalin A without causing cell agglutination or clustering of its receptors, induces some alteration in the conformation of receptor glycoprotein, which blocks the internalization of acidic sugar residues by subsequent irradiation.


An electrophysiological study on the effects of tryptophan and cortisol on schizophrenic and other mentally ill patient groups and on normal subjects. Direct current potentials measured on the scalp suprajacent to the midline prefrontal cortex appear to monitor the metabolic activity, via CO2 production, of this portion of the cerebrum. Changes in the frontal potential 90 min after oral administration of 2.0 g L-tryptophan and of 50 mg cortisol were studied in groups of subjects defined by age, sex, psychiatric diagnosis, and medication. Twelve normal males, 10 nonschizophrenic male psychiatric inpatients, and 6 normal females showed a significant decrease in their frontal voltages, most marked in the females, after tryptophan loading. A nonsignificant voltage increase was produced by subsequent administration of cortisol. Tryptophan-loading had an opposite, voltage-increasing effect, on the 30 schizophrenic subjects tested. This abnormal response was greatest in the male and postmenopausal schizophrenic subjects. Besides this sex effect, the abnormality increased with age up to a point, and was decreased by antipsychotic medication and cortisol. An explanation in terms of an abnormality in the relative hydroxylation of indoles in schizophrenic subjects is proposed.


Effect of midbrain raphe lesion on avoidance learning in aggressive mice. Lesion of the midbrain ventral raphe nucleus greatly improves the avoidance-learning efficiency of normal and isolated-aggressive Albino Swiss mice. Since the lesion of this brain area selectivity lowers the forebrain concentration of 5-hydroxytryptamine (5-HT), implications of the role of brain 5-HT in avoidance-learning acquisition are discussed.


Coping with stress: parallelism between the effects of septal lesions on growth hormone and corticosterone levels. In order to characterize the effects of a septal lesion on plasma growth hormone levels, blood samples were taken from normal and sham-operated rats and rats with a bilateral septal lesion under conditions of rest and following stressful stimulation. Nonlesioned control rats evidenced a significant base line diurnal rhythm in plasma growth hormone levels which was unaffected by a septal lesion. In response to stress, plasma growth hormone levels evidenced a significant drop. Minimum levels were reached 15 min after stimulation and remained depressed during 1 hr of observation after stimulation. Rats with a septal lesion evidenced the same pattern of growth hormone response to stressful stimulation as nonlesioned rats, however the magnitude of response was potentiated in septal rats. The present data were compared to the previously published corticosterone data from the same animals. A septal lesion has similar effects on boting 24-hour rhythm of either of these hormones but potentiated the response of both corticosterone and growth hormone to stress even though the direction of response is opposite. The results are interpreted as suggesting the septum is an element in some central coping mechanism which is involved when an organism reacts to environmental demands. Preliminary data further suggest that the same central coping mechanism is involved in both the behavioral and hormonal responses to stress.


Characteristics of unlimited access to self-administered stimulant infusions in dogs. When drug-naive dogs were given unlimited access to response-contingent intravenous infusions of either d-amphetamine, phenmetrazine, or methylphenidate, a regular cycle of drug intake interspersed with periods of voluntary abstinence was seen. During the drug self-administration phases there was a marked increase in locomotor behavior and stereotypy along with a decrease in body weight; the rest periods were characterized by minimal activity. These results are similar to those observed when humans engage in high-dose intravenous abuse of psychomotor stimulants.


[Adaptation in a detector system and differential sensitivity]. It is shown, that if during adaptation the sensitivity of single detectors of the models (1) is constantly decreasing proportionally to the level of their excitation, then the differential sensitivity of the whole  system to the stimulus near the adapting stimulus increase. The condition is deduced under which the perceived value of stimulus parameter is invarable during adaptation. Calculated from these conditions invariable values of line inclination, hue of the patch, are in agreement with experimentally obtained data.


[Spontaneous immuno-adherence in viral hepatitis]. Sera from patients with hepatitis, according to clinical and biochemical data, often give a spontaneous immune adherence (S.I.A.) with O human red blood cells. In 58% of the cases the test is positive and is associated with Australia (HBs) antigen in 6.7 to 9.5% of the patients. The titer cas reach 1/8192 and seems to follow the clinical evolution. S.I.A. also occurs at low titre (less than or equal to 1/32) in 18 to 30% of the cirrhosis tested, in 17 to 22% of various diseases and in 0 to 16% of different neoplasia. In spite of the fact that S.I.A. is not specific in its principle, it may be useful as a complementary test for detection and monitoring of viral hepatitis.


Growth kinetics of a yeast grown on glucose or hexadecane. The kinetics of growth of a Torulopsis sp. was investigated in a continuous culture with glucose or hexadecane as the carbon source; growth was limited by either carbon or nitrogen. The relationship between the concentration of the limited substrate and the steady-state growth rate of the organism was examined and tested against various models of growth. No existing model was found to describe the growth accurately and a new model has been proposed: (see article). It is postulated that this behavior would result from a simple first order reaction between the reactants of the rate-limiting enzymic reaction of the organism's metabolism.


Feather protein as a support for immobilizing enzymes. Feather meal protein was prepared in granular form and used as a support for lactase using glutaraldehyde as the crosslinking agent. The support gave a high retention of activity and in column operation it yielded apparent half-lives from 50 to 100 days. Because of its gel-like consistency (water content of about 90%), there is some diffusional restricting of activity as indicated by the kinetic data of soluble and immoblized enzymes.


Application of dynamic calorimetry for monitoring fermentation processes. The rate of heat evolution (kcal/liter-hr) in mycelial fermentations for novobiocin and cellulase production with media containing noncellular solids was measured by an in situ dynamic calorimetric procedure. Thermal data so obtained have proved significant both in monitoring cell concentration during the trophophase (growth phase) and in serving as a physiological variable in the fermentation process. The validity of this technique has been demonstrated by closing the overall material and energy balances. The maintenance energy in a batch fermentation can be calculated by integrating heat evolution data. This integration method is applicable to a fermentation lacking a precise cell growth curve. The maintenance coefficient, obtained for the novobiocin fermentation by Streptomyces niveus, is equal to 0.028 g glucose equivalent/g cell-hr. The production of novobiocin in the idiophase (production phase) also correlates well with the amount of energy catabolized for maintenance and this results in an observed conversion yield of glucose to novobiocin of 11.8 mg of novobiocin produced per gram of glucose catabolized. A new physiological variable, kilocalories of heat evolved per millimole of oxygen consumed, has been proposed to monitor the state of cells during the fermentation. This method may provide a simple way to monitor on-line shifts in the efficiency of cell respiration and changes in growth yields during a microbial process.


Observations on the pharmacology of cholinoceptive neurones in the rat brain stem. The pharmacology of spontaneously active cholinoceptive neurones in the brain stem of rats anaesthetized with urethane has been investigated using microiontophoresis to administer muscarinic and nicotinic agonists and antagonists. 2. Acetylcholine (ACh) excited most cells but occasionally depressed their activity. Muscarine, and the muscarinic agonists methacholine and bethanechol produced prolonged excitation or inhibition of cells whereas nicotine produced prolonged excitations but no inhibitions. 3 Atropine selectively antagonized ACh excitations and both excitation and inhibition of neuronal activity produced by muscarine and muscarinic agonists, but not the excitations produced by nicotine, glutamate or DL-homocysteic acid. 4 Dihydro-beta-erythroidine (DHBE) and tubocurarine antagonized both ACh and nicotine excitations but not those induced by glutamate or DL-homocysteic acid. Inhibitions by ACh or muscarine were not affected. 5 It is concluded that excitations of cholinoceptive neurones in the rat brain stem may be mediated by activation of both muscarinic and nicotinic receptors whereas inhibitions are mediated by activation of a muscarinic receptor.


Hand preference patterns in psychiatric patients. A brief survey, using a standardized questionnaire, has been made of hand preference patterns in a series of patients admitted to hospital with functional psychiatric disorders. A small but significant shift towards left hand preference has been observed, more marked in the young patients and virtually confined to the males. Psychotic patients show such a shift more clearly than neurotic or personality disordered patients. Simple genetic factors do not appear to be responsible for these sinistral tendencies. The findings are discussed in relation to possible acquired abnormalities of functional brain organization.


[Chemical and enzymatic degradation of Helminthosporium spiciferum cell walls. Protective role of pigments]. Cell walls of the fungus Helminthosporium spiciferum, prepared by mechanical grinding of the cells, were treated chemically and with helicase. The purified cell walls and the fractions were analyzed chemically and micromorphologically to localize the major cell-wall constituents and to determine the protective role of the pigments against degradative processes. The encrusting of the external area by the pigments associated with chitin formation leads to an increased resistance of this cell-wall area against the degradative processes.


Conjugation in the yeast Guilliermondella selenospora Nadson et Krassilnikov. Conjugation in Guilliermondella selenospora took place via conjugation tubes from cells in different hyphae. Afterwards, a septum was formed in the channel connecting the two cells which turned into asci or formed buds which became asci. Conjugation between adjacent hyphal cells was also found. Cell contact without fusion with denticles and stalks, which occurs in G. selenospora, was compared with conjugation.


The effect of light on acid-soluble polysaccharide accumulation in Sclerotium rolfsii Sacc. A light-stimulated increase in beta-1,3 glucan accumulation was observed for Sclerotium rolfsii Sacc. The acid-soluble polysaccharide accumulated in large quantities in 'white' light- and blue light-grown cultures. This polysaccharide also accumulated in both dark- and red light-grown cultures as well. However, the quantities were significantly lower when compared to the 'white' light- and blue light-grown cultures. A greater quantity of polysaccharide accumulated in red light-grown cultures than in dark-grown cultures.


Some comparative observations on the relative contributions of alternate pathways in the metabolism of glucose by Candida utilis. Candida utilis was grown in batch, chemostat, and continuously synchronised (phased) culture on a nitrogen-limited glucose mineral salts medium: phosphorus- and carbon-limited phased cultures were also used. The 14CO2 evolved from [G-1-14c]and [G-6-14c] was used, as a simple C1/C6 ratio, to observe the relative changes in EMP and HMP contributions during growth of the cultures. The ratio varied during the cell cycle, and changed with growth rate, and with nutrient limitation. The changes generally indicated that the HMP predominated, most notably in the early part of the batch-growth sequence and early in the cell cycle. The overall results reflected the relative merits of the different cultivation techniques for examining microbial metabolism: the advantage of a greater resolution by the synchronised method, based upon the unit performance rather than the randomised mean performance of the cell population...


The ultrastructure of the Spilocaea state of Venturia inaequalis in vivo. There are indications that the fungus enzymatically degrades the cuticle and epidermal cell wall. The epidermal cells and to a lesser degree the palisade mesophyll cells beneath a sporulating lesion (susceptible reaction) are killed or seriously disrupted. Various stages of conidiogenesis, including development of the pprimary conidium, were observed. A conidium is delimited by a two-layered transverse septum. Before condium secession, a new two-layered inner wall is laid down around the entire conidiogenous cell adjacent to the plasmalemma. The apical region of the new inner wall proliferates beyond the annellation scar left by the seceded conidium and eventually produces another conidium.


A technique for the differential isolation of hop mosaic virus and hop latent virus. Two rod-shaped viruses of similar length occurring in hops were isolated from a mixed infection. One (hop mosaic virus) infected Nicotiana clevelandii, produced mosaic symptoms in mosaic-sensitive hop cultivars, and did not infect Chenopodium murale. The other (hop latent virus) infected C. murale but not N. clevelandii and did not produce mosaic symptoms in hops. Neither N. clevelandii nor C. murale developed distinct symptoms after infection with the respective virus. Both viruses produced a systemic chlorotic flecking in Cluster hop seedling clones.


Phagocytic and chlamydiae-inhibiting activities of stimulated and nonstimulated periotneal mouse macrophages. Phagocytic and chamydiacidal properties of nonstimulated and stimulated mouse mononuclear cells for two Chlamydia psittaci 6BC strains were investigated. It was determined that macrophages kept in monolayer culture (i.e. stimulated phagocytes) developed much more efficient chlamydiacidal ability than did cells kept in suspension directly after harvest (i.e. nonstimulated phagocytes). A thousandfold decrease of chlamydial infectivity was observed 60 min after induction of phagocytosis in stimulated macrophages, irrespective of the strain offered. In contrast, the infectivity of both chlamydial strains remained essentially unchanged up to 1 h after phagcoytosis by nonstimulated phagocytes. Electron-microscopic examination indicated that chlamydiae were disintegrated within phagolysosomes of stimulated phagocytes within minutes after phagocytosis, although ultrastructurally altered chlamydiae not enclosed in vacuoles were also observed in the cytoplasm of several mononuclear phagocytes at different times after phagocytosis.


The oxidation of terminal D-galactofuranose residues of a galactan and a glycoprotein by a D-galactose oxidase preparation from Dactylium dendroides. A galactan, isolated from the unicellular organism Prototheca zopfii, and a glycoprotein from a hyphal cell-wall fraction of the fungus Pithomyces chartarum have been oxidised by a D-galactose oxidase preparation from Dactylium dendroides. The oxidised polymers were subsequently reduced with sodium borotritide. The site of oxidation was identified as C-6 of non-reducing D-galactofuranosyl residues in both polymers.


UDP-N-acetyl-D-glucosamine-asparagine sequon N-acetyl-beta-D-glucosaminyl-transferase-activity in human serum. Serum contains a sugar transferase which is able to catalyse the glycosylation in vitro of the asparagine residue present in the sequence Asn.Leu.Thr in bovine pancreatic ribonuclease. UDP-2-Acetamido-2-deoxy-D-glucose (UDP-N-acetyl-D-glucosamine) acts as a donor, although the mechanism of the transfer is unexplored. Spermidine and Mn2+, as well as CDP-choline, can act as activators for the reaction. Monoglycosylated ribonuclease (ribonuclease-GlcNAc) has been separated (23% yield) from unreacted ribonuclease A by affinity chromatography on a column of wheat-germ agglutinin bound to Sepharose, and characterised. A possible reason for the presence of the enzyme in serum is suggested.


Isolation and some properties of DNA coding for tRNA1met from Xenopus laevis. DNA containing the reiterated genes for tRNA1met has been partially purified from Xenopus laevis by centrifugation in actinomycin C1-CsCl and Ag+-Cs2SO4 gradients. These gradients separate the tRNA1met genes from those coding for tRNA2met and tRNAval, thus confirming our earlier suggestion that these genes are not intermingled with each other (Clarkson, Birnstiel, and Purdom, 1973a). The gradients also demonstrate the existence of a minor 5S DNA fraction which appears to differ from that previously isolated by Brown, Wensink, and Jordon (1971). When the enriched tDNA1met is digested to completion with either of the restriction endoncucleases EcoRl or Hpa l, the tRNA1met genes are predominantly found within DNA fragments that are about 3100 base pairs long. A partial digestion with EcoRl shows that these fragments arise from the regular spacing of the enzyme restriction sites. The 3100 base pair EcoRl fragments are cleaved by Hpa l into fragments to two size classes, one of which is about 2200 base pairs long and contains the tRNA1met genes. The shorter fragments are about 700 base pairs long, and they appear to contain genes coding for at least one other kind of tRNA species. X. laevis tDNA1met thus comprises tandemly repeated DNA whose component parts show little if any length heterogeneity.


A novel temperature-sensitive mammalian cell line exhibiting defective prophase progression. A temperature-sensitive mammalian cell line has been isolated which grows and divides normally at the permissive temperature of 33 degrees C. When incubated at 39 degrees C, the nonpermissive temperature, interphase cells continue to enter a prophase-like state. Chromatin-like material condenses and coalesces into dark-staining clumps rather than into discernible chromosomes. Disappearance of the nuclear boundary is observed, but re-formation of the boundary around the clumps fails to occur. In corporation of labeled precursors reveals a decrease in protein synthesis which is accompanied by a slower decrease in DNA synthesis. Approximately 0.2% of the mutant cells revert in their capability of growth and cell division at 39 degrees C. These "revertants" are found to contain a higher number of chromosomes. The isolation of this mutant is based on the initial observation that the cells become rounded at the nonpermissive temperature. The cell-rounding process characteristic of mitotic cells should serve as a useful marker in the isolation of mitotic mutants.


Translation of Xenopus liver messenger RNA in Xenopus oocytes: vitellogenin synthesis and conversion to yolk platelet proteins. Xenopus liver vitellogenin and albumin mRNAs injected into Xenopus oocytes are correctly translated, as shown by specific immunoprecipitation and co-electrophoresis with purified Xenopus vitellogenin (molecular weight 210,000 daltons) and albumin (molecular weight 72,000 daltons). Vitellogenin made in oocytes under the direction of injected liver mRNA is unstable compared to other proteins made on injected messengers (such as albumin and globin) and endogenous oocyte proteins (including actin), the half-life of newly made vitellogenin being about 8 hr. Pulse-chase experiments with 35S-methionine show vitellogenin to be a precursor to yolk platelet lipovitellin (molecular weight 120,000 daltons), while 3H-serine labeling demonstrates conversion to phosvitin (molecular weight 34,000 daltons). In contrast, injected 3H-serine 35S-methionine-labeled Xenopus vitellogenin protein is not converted to yolk platelet proteins and is degraded rather slowly (half-life, 23, 29 hr). Phosphorylation of serine residues in phosvitin can be detected in oocytes injected with 32PO4 or gamma-32P-ATP; thus exogenously derived yolk platelet protein is further modified, or turned over, once it is within the oocyte. Moreover, vitellogenin made in oocytes programed with liver mRNA is phosphorylated. Thus phosphorylation, assembly into yolk platelets, and cleavage are events that do not require vitellogenin supplied by the normal pathoways involved in yolk formation (synthesis and post-translational modification in the liver, transport in the serum, and follicle cell-dependent pinocytosis). Vitellogenin mRNA sediments at about 29S in a sucrose-SDS gradient, while albumin messenger peaks at 16S; both species contain poly(A). These liver mRNAs are functionally stable in oocytes for at least 5 days. Vitellogenin-forming activity, relative to albumin, actin, or total endogenous activity, increases with time, and the final rate of 2-2.5 times the initial rate is only reached 3 days after injection. The potentiation effect probably stems from an increase in the efficiency of translation of vitellogenin mRNA. The availability of homologous mRNAs now permits injected messenger to be used as a valide probe of oocyte function; the biological activity of mRNA from a non-ovarian Xenopus tissue proves that some at least of the translational systems within the Xenopus oocyte are not cell type-specific. Moreover, the whole cell system is eminently suitable for assaying putative translational (and possibly transcriptional) control elements from frog liver.


Stimulation of DNA synthesis by ouabain and concanavalin A in cultures of embryonic neural retina cells. Ouabain and concanavalin A, agents which bind to specific sites in the cell membrane, stimulate DNA synthesis and cell replication in monolayer cultures of neural retina cells from late chick embryos. The results suggest a relationship between control of retina cell replication and properties of the cell membrane. The experiments involved measurements of 3H-thymidine incorporation in primary monolayer cultures (24-48h) of retina cells from embryos of different ages. Stimulation by ouabain was greatest in cells from 14-day embryos, and its magnitude was similar to that elicited in these cell cultures by concanavalin A. Simultaneous treatment of 14-day retina cells with both agents resulted in a greater than additive stimulation of DNA synthesis. Our results demonstrated that, although during normal embryogenesis cell replication in the neural retina has virtually ceased by day 14 of development, some cells retained a capacity for mitogenesis when exposed to conditions such as provided in these experiments. By autoradiography the responding cells were identified as large epithelioid retina cells (LER cells). Under optimal conditions of simultaneous treatment with ouabain and Con A about 20% of the LER cells showed stimulation of DNA synthesis. The nature of LER cells and other aspects of our findings are discussed.


Subpulmonary obstruction in congenitally corrected transposition of the great arteries due to ventricular membranous septal aneurysms. The clinical, hemodynamic, and angiographic observations, as well as the surgical approach used for repair in three patients with congenitally corrected transposition of the great arteries and ventricular membranous septal aneurysms, are presented. In two of the three patients the membranous septal aneurysm caused subpulmonary obstruction, with 94 and 125 mm Hg systolic gradients. In each patient the aneurysm was demonstrated by angiocardiography, which also showed differences in size and shape with cardiac systole and diastole. Review of the previously described reports indicates that patients with congenitally corrected transposition often display various forms of pulmonary outflow obstruction and when a ventricular membranous septal aneurysm exists, a significant subpulmonary obstruction is present in most patients. The unique anatomic relationship between the pulmonary artery and a ventricular membranous septal aneurysm in patients with transposition of the great arteries with and without atrioventricular discordance explains why subpulmonary obstruction sometimes develops.


Legislation relevant to perinatal care.

Legislation relevant to perinatal care. The legislative process is one route to follow in the attempt to change and improve perinatal care. Payment by the State Crippled Children's Service for medical care of certain costly high-risk neonatal conditions, only to qualified specialists and in centers meeting acceptable standards has had a snowball effect on upgrading neonatal care in this state. Not only has a large network of neonatal care centers and infant transport systems been developed, but there has been a rush especially on the part of nurses, to get special training in neonatal care. This has included not only the care of the sick neonate, but a look at newborn evaluation and resuscitation in the delivery and newborn areas. It is expected that this same center development and education and training process will now be extended to obstetrical care, as there is renewed interest in special care for high-risk mothers because of Assembly Bill 1326. The new hospital perinatal regulations mandate improvement of care in community hospitals where the majority of deliveries take place. The emphasis is on a larger and better educated staff, more concern with patients rights, and provision of a more humanistic family centered care as well as continual evaluation of maternal and neonatal outcome. The greatest limitation has been lack of Health Department staff to provide adequate consultation and surveillance of these services for compliance with the new laws. There has been an approximate 10 per cent reduction in the number of hospitals with maternity services- from 416 in 1968 to 369 today. While much of this consolidation may have been due to the fall in birth rate, these regulations have also contributed to the process. Most important of all, these laws have kept perinatal care constantly in the consciousness of California health care providers and consumers.


Indirect evidence of calcitonin secretion in man. 1. The effect of calcitonin, a large amount of calcium given orally, pentagastrin and glucagon on plasma 47Ca radioactivity curves in subjects pretreated with 47Ca was examined. 2. A sudden increase of plasma radioactivity after intravenous administration of calcitonin and pentagastrin and after the oral calcium load was observed in normal subjects; the intravenous infusion of glucagon was less effective. 3. Two thyroparathyroidectomized patients who responded to the calcitonin infusion did not respond to the oral calcium load. 4. These data may be considered to offer indirect evidence of endogenous calcitonin secretion in man.


Subpulmonic stenosis as a result of noncalcific constrictive pericarditis. The clinical, electrocardiographic, and radiologic features in a patient with a pericardial band which produced an unusual form of infundibular pulmonary stenosis are presented. The findings are unique in that the band was not calcified and, therefore, not visualized roentgenographically and developed without a previous history of pericarditis or pericardial surgery. The diagnostic value of cineangiography is stressed and illustrated.


[New arteriovenous fistula for extracorporal hemodialysis using a cattle-artery heterologous graft]. 17 patients are reported in whom an arterio-venous shunt has been established by means of an arterial graft (bovine origin). The indications were: need of maintenance hemodialysis under absence of functioning shunt and lack of peripheral vessels suitable for construction of a Cimino fistula. 14 shunts developed function without any complication. In 2 patients thrombosis of the graft occured. Both of them could be re-established in function by thrombectomy. 2 cases necessitated removal of the transplant. The question as to an immunological rejection remains open up to now. The advantages of the new shunt are: immediate readiness for hemodialysis, sufficiently long distance for puncture, easy and painless punctures and the possibility of access to deeper vessels.


Ultrastructure of segmentation mitoses in cleaving newt eggs after blocking of centrospheres by quinoline. In quinoline treated blastomeres of Triturus helveticus Raz. and Pleurodeles waltlii Michah., microtubules disappear and around the centrioles markedly enlarged dense bodies accompanied by striated bodies are accumulated, from prophase beyond metaphase. It is concluded that these bodies in untreated cells play a role in the formation of the spindle microtubules. - During metaphase the chromosome are smooth-surfaced and show a pronounced tendency to stickiness. During ana-telophase they become surrounded by nuclear membranes and form caryomeres. - Qinoline does not interfere with centriole replication, but prevents the separation if diplosomes that have been already formed, if it acts before that separation. - Some centrioles exhibit different degrees of ultrastructural disarrangement.


Affinity chromatography of collagen glycosyltransferases on collagen linked to agarose. Denatured citrate-soluble collagen was coupled to agarose by the cyanogen bromide activation technique, and columns prepared from this material were studied for affinity chromatography of collagen glycosyltransferases. Both collagen glycosyltransferases became bound to the column, the degree of binding and the capacity of the column being higher with the glucosyltransferase than with the galactosyltransferase than with the galactosyltransferase. The addition of Mn2+ enhanced the binding, especially with the glucosyltransferase. The enzymes were eluted from the column with small peptides prepared from collagen, and they were separated from the peptides by gel filtration. With this procedure a collagen glucosyltransferase purification of about 5000-fold and a collagen galactosyltransferase purification of about 1000-fold was obtained from chick embryo extract by relatively simple steps.


[Preparation and partial physico-chemical characterization of sheep-serum transferrin]. Sheep-serum transferrin shows marked polymorphism and more than 20 alleles have been identified although only 4 or 5 of these have a frequency higher than 1%. Each of the alleles has two bands in starch-gel electrophoresis, corresponding to a major and a minor fraciton. This paper describes the isolation and partial characterisation of the two fractions from the transferrin of sheep homozygous for Tf B. The purification consisted of: (a) precipitation by ammonium sulphate, (b) chromatography on CM-cellulose and, finally (c) chromatography on DEAE-Dephadex. The purification procedure had no effect on the electrophoretic mobility of the two fractions and they appeared homogeneous by starch-gel and polyamide-gel electrophoresis and by immuno-electrophoresis. The amino-acid compositions of both fractions were very similar and the sequences of the first eight amino-acid residues: Ser-Pro-Glu-Lys-Thr-Val-Arg-Trp- were identical for both bands. These results, and the fact that the two fractions are found in all genetic variants and always have the same relative mobility strongly suggest that the differences do not lie in the polypeptide chain. From the results of hydrolysis by neuraminidase and assay of sialic acid, the major and minor fractions most probably contain two and three sialic acid residues (exclusively N-acetyleuraminic acid) respectively, thus explaining the different electrophoretic mobilities. The sialic-acid content has been calculated on the basis of a molecular weight of 77500 as determined both by low-speed equilibrium ultracentrifugation and by Archibald's method...


Kinetics of refolding and reactivation of rabbit-muscle aldolase after acid dissociation. Tetrameric rabbit muscle aldolase has been dissociated to the monomer at pH 2.3 and fully reassociated and reactivated at pH 7.6 Kinetics of reactivation and refolding were followed by slow and fast kinetic techniques after neutralization of the acidic enzyme solutions by dilution or rapid mixing. Rate constants (k), reaction orders (n), and activation evergies (E) were calculated from measurements on the time, concentration, and temperature dependence of the reactions. The experimental results prove reactivation at high enzyme concentration (c greater than 4mug/ml) to obey first-order kinetics; at lower concentrations a transition to a higher reaction order is observed. Because of concentration-dependent deactivation at low enzyme concentrations reactivation measurements could not be extended below c approximately o.5 mug/ml. In the accessible concentration range incomplete separation of reassociation and  transconformation processes as well as intrinsic residual activity of the isolated subunits lead to an average value of n = 1.40 +/- 0.18. Renaturation as monitored by protein fluorescnce is a multi-step process composed of a fast increase in flurescence emission (first-order rate constant k approximately  15 s-1) and a slow concentration-dependent decrease which parallels the recovery of enzyme activity (n = 1.46 +/- 0.12). The activation energy of both processes is of the order of E = 12-16 Kcal/mol (50-67 kJ/mol). Reassociation is a prerequisite of full catalytic function and native fluorescence.


The kinetics of hydrolysis of some extended N-aminoacyl-l-lysine methyl esters. 1. The action of two active forms of bovine trypsin (alpha and beta-trypsin) on a series of specific methyl ester substrates of general formula: N-acetyl-(glycyl)n-L-lysine methyl ester (n = 0, 1, 2) and N2-benzoyl-L-arginine ethyl ester have been investigated. With the L-lysine methyl esters the catalytic rate constant for hydrolysis (kcat) was found to be significantly lower for alpha-trypsin than for beta-trypsin, whereas with N2-benzoyl-L-arginine ethyl ester there was no significant difference for the two enzymes. 2. By measurement of the kinetic constants (kcat and Km) in the presence of a nucleophile, which competes with water in the deacylation process, it has been shown that, in common with the specific ester substrates of trypsin, the rate-determining step for the extended L-lysine methyl esters is decaylation of the enzyme. 3. It has been found that by extending the aminoacyl group of N-acetyl-L-lysine methyl ester by one glycine residue (n = 1), a greatly enhanced deacylation rate constant is observed for both alpha and beta-trypsin. The higher rate constants were maintained at the higher levels by the addition of a further glycine residue (n = 2). These results have been interpreted in terms of the 'induced fit' hypothesis the substrates binding to an enzyme subsite adjacent to the active site. 4. The beta-trypsin-catalysed hydrolysis of the L-lysine substrates was investigated over a range of temperature (15--35 degrees C). The Arrhenius law was obeyed, within experimental error, by all three substrates allowing the estimation of the thermodynamic function of activation (delta S not equal to and deltaH note equal to) for the deacylation reactions. The significantly higher values of deltaS not equal to and deltaH not equal to obtained for the two extended substrates are interpreted in terms of additional hydrogen bonding between the longer aminoacyl chains and the enzyme molecule. The results are compared with those for non-extended specific substrates, which have a possible hydrophobic interaction with the enzyme surface.


Wireless endoscopy.

Wireless endoscopy. A completely new and different principle of 'wireless' endoscope is described. The 'contact endoscope' consists of two parts, a rod which is the optical element and a light recovery jacket. This endoscope utilizes the surrounding light, either daylight or artificial light. It allows endoscopic photography and cinematography with a magnificant of nearly five. For urethrocystoscopy it represents a safe introductory rod (mandrel) introduced under visual control. It can be used in kidney surgery for finding stones in calyxes.


Differential attenuation of some effects of haloperidol in rats given scopolamine. In a series of pharmacological tests the ability of 0.6 mg/kg of the anticholinergic scopolamine to decrease the effectiveness of the neuroleptic heloperidol varied widely. Most severely attenuated was production of catalepsy followed in order of decreasing interference by inhibition of amphetamine-induced rotation, inhibition of amphetamine- or apomorphine-induced stereotyped behavior, inhibition of conditioned avoidance responding and lastly attenuation of the haloperidol-induced increase in striatal HVA. By use of such a relatively low dose of scopolamine the behavioral effects of heloperidol were dissociated from effects on dopamine turnover in the striatum. If behavioral tests in animals can be related to the clinical effects of neuroleptic drugs, those effects of haloperidol severely reduced by scopolamine may be related to extrapyramidal effects.


Plasma free and total tryptophan during the oestrus cycle, in ovariectomized and in male rats. Plasma free tryptophan was higher during prooestrus and early oestrus than at metoestrus or in ovariectomized or male rats. In contrast, total tryptophan was higher in ovariectomized and male rats than at any time in cycling females.


[Clinico-pathological study of chronic thyroiditis (author's transl)]. Although chronic thyroiditis has been recently noticed as an autoimmune disease, the histological classification based on clinico-pathological correlation is still vague at present. The specimens subjected to this study were taken by exploratory excision from 63 patients who were clinically diagnosed as chronic thyroiditis. Histological observation was done regarding the progress of pathologic changes in the thyroid gland. Lymphoid type (14 cases): lymph-follicle formation with scattered infiltration of lymphocytes was the predominant feature in this group. No definite alteration of the thyroi- epithelium was observed. Some of the so-called focal thyroiditis may be included in the early stage of this group. Some of them may develop into tremendous proliferation of lymphoid tissue in its late stage. Oxyphilic cell type (34 cases): oxyphilic alteration of the epithelium was predominant, but lymphocytic infiltrations observed in varying degrees was not an essential feature of this group. Fibrosis may possibly become more prominent with destruction of thyroid follicles in the late stage. Hyperplastic type (9 cases): most follicles were lined by cuboidal or tall epithelium with papillary infolding and contained thin colloid and some cell aggregates. This type of thyroiditis also may result in a proliferation of interlobular fibrous tissue in its more advanced stage. Fibrous type (6 cases): this type of thyroiditis is believed to be a late stage of the other three types. The original type of thyroiditis, however, was not clear because of advanced fibrosis. Evidences of the progress of the histological changes were observed in 12 out of 40 patients in whom the specimens were taken from both lobes and in two others whose specimens were taken twice at different stages. Clinical correlation of this classification was studied. The lymphoid type was found in 14 out of 53 female patients, but in none of 10 males. In age distribution the hyperplastic type was observed to be more common than the other types among younger people. The clinical signs of hypothyroidism were recognized most frequently in the oxyphilic cell type but not in the hyperplastic type. Thyroid function test, however, demonstrated no definite differences among the four types. Relationships were discussed in this paper between this author's classification and that of Woolner--also relationships to Hashimoto's thyroiditis and Riedel's struma.


[Rheumatoid factor in diabetes mellitus (author's transl)]. This study was undertaken to try to determine the incidence of rheumatoid factor in patients with diabetes mellitus by the Hyland RA slide test and the RAHA test (the sensitized sheep red cell agglutination test). Sera from 507 patients with diabetes mellitus and 2073 controls were investigated for the presence of rheumatiod factors. RA test was positive in 8.7 per cent and RAHA test in 6.1 per cent in sera from patients with diabetes mellitus compared to 4.1 percent and 3.6 per cent respectively in healthy subjects.


[Studies of binding of testosterone-estradiol binding globulin for estradiol-17beta (author's transl)]. A fundamental study of testosterone-estradiol binding globulin (TeBG) activity for estradiol-17beta was investigated by equilibrium dialysis. In addition, the interaction of TeBG activity for estradiol and estradiol metabolism during a menstrual cycle and in hyperthyroidism was attempted to elucidate. Estradiol in plasma and 24 hrs urine was measured by radioimmunoassay. TeBG binding capacity (TBC) was measured by ammonium sulfate method and associate constant (Ka) of TeBG for estradiol was estimated by equilibrium dialysis. 1. In several pooled plasmas, the mean Ka of TeBG for estradoil was 1.25 X 10(8) L/M at 4 degrees C and 0.78 X 10(8) L/M at room temperature. The Ka decreased with increasing temperature and could not be determined with a sufficient accuracy at 37 degrees C. The mean TBC was 4.4 X 10(-8) M/L. 2. TBC and Ka of TeBG for estradiol during the normal menstrual cycle of a woman were fairly unchanged. The unconjugated estradiol value in urine showed the same biphasic pattern with a midcycle peak as that of the unconjugated estradiol value in urine. 3. A normal man was induced hyperthyroidism with a administration of triiodothyronine. The treatment of triiodothyronine caused a remarkable increase of TBC, but no changes of Ka of TeBG for estradiol and unconjugated estradiol values in plasma and urine. From these results it was suggested that the rate of metabolism of estradiol increased in hyperthyroidism.


[Preovulatory changes of steroidogenesis in isolated rabbit follicles (author's transl)]. In an attempt to investigate the effect of ovulating hormone on the steroidogenesis of mature follicles in the course of ovulation, transitory changes of steroidogenesis in isolated rabbit follicles have been studied at several intervals after injection of an ovulatory dose of human chorionic gonadotropin (hCG). Five to ten follicles of approximately 1-2 mm in diameter were isolated from ovaries of a mature rabbit (2.5-3.0 kg) under streomicroscope, before and at the 3rd, 6th, 9th and 12th hours after intravenous injection of of 100 IU/kg of hCG. Follicles were incubated with 100 muCi of acetate-1-14C in 2 ml of Krebs-Ringer bicarbonate buffer (pH 7.4) at 37 degrees C for 3 hours under 95% oxygen plus 5% carbon dioxide. Each incubation was terminated by quick freezing and stored forzen at -20 degrees C until eighty follicles had been collected for each time period before commencement of analysis. Incorporation of radioactive acetate into pregnenolone, 17-hydroxypregnenolone, progesterone, 17-hydroxyprogesterone., 20 alpha-dihydroxyprogesterone, dehydroepiandrosterone, androstenedione, testosterone, estrone and estradiol-17beta were analysed by the reverse dilution technique and identified in radiochemically pure form by recrystallization to constant specific activities. The steroidogenic activity of the follicles was evaluated by overall as well as fractionated incorporations. A peak in the overall incorporation of 14C- acetate into the ten steroids at the 3rd hour after hCG injection, followed by gradual decrease up to the 9th hour was observed. The incorporation decreased markedly to a minimum level at the 12th hour after hCG injection, which was below the level of preinjection control. Comparable quantitative fluctuations were found with the fractionated incorporation of 14C-acetate into the C21 and C18 steroids in the time sequence following hCG injection. However, the fractionated incorporation into C19 steroids reached to a maximum at the 6th hour after hCG injection. 5istribution patterns of incorporation among the individual steroids were varied at each interval of time. In the non-injected control, mature follicles synthesized predominantly estradiol-17beta, testosterone and androstenedione. Divergent steroids were formed from radioactive acetate at the 3rd hour after hCG injection. These included porgestogen, androgen and estrogen, but pregnenolone and 17hydroxyprogesterone were the two principal steroids produced. There was no essential difference in the steroidogenic patterns between the 6th and 9th hour, the major products being C21 and C19 steroids such as pregnenolone, 17hydroxyprogesterone, dehydroipiandrosterone and testosterone. The three androgens were the major steroids formed at the 12th hour after hCG injection. Thus the chages in the steroidogenic profile of the follicle was obvious in the course of ovulation. The basis of qualitative changes in follicular steroidogenesis during the process of ovulation have been discussed in connection with an accompanying effect of an ovulatory dose of hCG.


[Thyroglobulin and microsomal antibodies in diabetes mellitus in childhood (author's transl)]. The purpose of our study was to determine the incidence of thyroid antibodies in diabetes mellitus in childhood and to discuss the correlation between thyroid antibodies and insulin antibodies. The sera of 50 children with diabetes mellitus and 437 children as disease controls were tested by thyroglobulin and microsome-coated tanned red cell hemagglutination test (Fuji Zoki Co. Tokyo). One of the 50 children with diabetes mellitus (2%) was positive with antithyroglobulin antibodies and eleven (22%) were positive with antimicrosomal antibodies compared to 0.4% and 1.1% respectively in 437 disease controls. To clarify the association of insulin antibodies and thyroid antibodies in diabetes mellitus in childhood, insulin antibodies were demonstrated by using a modified method described by Wright. One of the 33 children with negative insulin antibody (2.8%) was positive with antithyroglobulin antibodies and eight (24%) were positive antimicrosomal antibodies. No evident correlation was observed between antithyroid antibodies and insulin antibodies.


[Further studies on serum calcium and inorganic phosphorus in postmenopause (author's transl)]. The author previously reported that the serum calcium (Ca) and inorganic phosphorus (P) levels were gradually decreased with advance in age in regularly menstruating women, but that they were rapidly increased soon after the menopause. In this experiment the changes of the serum Ca and P levels in postmenopausal women were studied more precisely in 410 peri- or post-menopausal subjects and 534 controls with regular menses. They were classified into 12 groups according to age or period since last menses. The clavicular cortical thickness was also measured as an index of bone resorption. Blood samples were taken between 9-11 A.M. after an overnight fast and the separated sera were applied to a Technicon Autoanalyzer SMA 12/60 and analyzed. The mean levels of Ca and P in serum were both increased following the menopause and reached a plateau in Groups F-3 (12-24 months since last menses) and G (2-5 years since last menses). In Group G (5-10 years since last menses) both the levels were slightly declined. Then a dissociation was observed in Groups I-1 and I-2. This was a successive decrease of Ca and a re-increase of P. This dissociation might be explained partly by the physiologic fall of parathyroid function. Furthermore the clavicular cortical thickness was measured with a micrometer in the middle of the clavicle on the chest X-ray film of the same subjects and the clavicular score was computed as Cortical width X 100/Whole width. The score was gradually but steadily reduced with the lapse of time after the menopause and in Group I-2 it was lowered to approximately 45% of that in the premenopausal groups. From these results it is certain that the decline of estrogen secretion led to the elevation of the serum Ca and P levels and subsequently to the reduction in the clavicular score.


[Microdetermination of TSH in dried blood spot--its use in the mass-screening for congenital or juvenile primary hypothyroidism (author's transl)]. Blood TSH (Thyroid Stimulating Hormone) was successfully measured by radioimmunoassay in a dried blood spot on filter paper which is obtained in newborn screening for metabolic disorders. By this method, the minimal detectable level of blood TSH was about 10 muU/ml, which is the approximate upper limit of normal values of blood TSH. Good correlation was found between the TSH values obtained from this specimen and from liquid serum samples from the same subjects. The duplication of assay of a single sample was not necessary. A screening program using the method described here was initiated and a 4 year old infant was found to have primary hypothyroidism, later verified by other testing methods. Since the technique is simple and adequately sensitive for the detection of hypothyroidism, it could be a valuable method for use in mass-screening of newborns for congenital hypothyroidism.


Food availability as a determinant of the rats' circadian rhythm in maternal behavior. Thre present study was designed to determine whether environmental factors other than light would alter the dialy periodicity in the rat's maternal behavior and whether such a change in the dam's behavior pattern would in turn alter the pattern of her sucklings' milk intake. Experiment 1 determined the daily pattern of litter weight gain (which presumably reflects milk intake) displayed by young reared by dams fed and watered either ad lib (Group C), only during the day (Group D), or only at night (Group N). Experiment 2 compared nocturnal and diurnal frequency of maternal behavior in Groups C, D, and N. The pattern of both litter weight gain and maternal behavior was observed to be diurnal in Groups C and N and nocturnal in Group D.


Maternal hyperphenylalaninemia: an experimental model in rats. Experimental maternal hyperphenylalaninemia produced in pregnant F344 rats by the combined use of p-Chloro-DL-phenylalanine and L-phenylalanine reduced fetal birth weight in comparison to saline-injected controls. Offspring who experiences hyperphenylalaninemia in utero died within 5 days after birth. Fetal plasma phenylalanine levels were several times higher than maternal plasma phenylalanine levels, indicating that the placenta actively concentrates maternal phenylalanine. Fetal brain phenylalanine levels rose in direct proportion to elevations in fetal plasma phenylanaline, whereas maternal brain phenylalanine levels remained low during maternal plasma phenylalanine elevation; the contrast suggests that the maternal brain is better able than the fetal brain to screen itself against high circulating plasma phenylalanine levels.


The effects of prenatal psychological stress on the sexual behavior and reactivity of male rats. Male offspring of prenatally stressed rats showed low levels of copulatory behavior during a series of brief tests with estrous females but successfully impregnated female cagemates during long term tests of breeding effectiveness. Sexual performance on the short term tests was inversely related to open field activity with the offspring of prenatally handled mothers exhibiting more sexual behavior and less open field activity. The results support the notion that prenatal stress influences both the sexual behavior and emotional reactivity of male offspring. Deficits observed during short term tests of breeding effectiveness appear to be reversible under optimal conditions.


A longitudinal analysis of maternal aggression in Rockland-Swiss albino mice. Female Rockland-Swiss albino mice were treated for aggression during 6 successive pregnancies and lactation periods or until they ceased to re-mate. Fighting behavior was limited only to the lactation phase of each reproductive cycle. Although most animals exhibited fighting during every lactation period they completed, some exhibited fighting in some lactation periods but not in others. The intensity of aggression, as measured by the time spent fighting, was highest during the beginning of each lactation period and declined by the end of each period. Moreover, the intensity of aggression increased across the first 3 lactation periods and then declined on later lactation periods to a point below initial levels. Finally, the fighting behavior exhibited by multiparous animals was not due simply to previous fighting experience in that some multiparous mice exhibited postpartum aggression on the 6th lactation period in the absence of fighting experience during earlier lactation periods.


Hypothermia causes adult-like retention deficits of prior learning in infant mice. Nine-day-old S-W mice receiving deep body hypothermia or hyperthermia immediately after escape training were retested 1 or 24 hr later. Results indicated that hypothermia impaired 24-hr retention but had no effect upon 1-hr memory. Hyperthermia had no effect, with the mice demonstrating retention of the escape response at both retest intervals. In Experiment 2, administration of hypothermia or hyperthermia 23 hr after original training had no effect upon memory nor did either treatment produce motoric deficits upon retest 1 hr following thermal exposure. Experiment 3 indicated that hypothermia administered immediately after training produced retention deficits directly related to amount of body temperature reduction following hypothermia. These data are similar to those obtained with adult mice and suggest that memory processes occurring in 9-day-old mice may represent the onset of functioning of processes underlying adult long-term memory.


Genetic models for the analysis of data from the families of identical twins. Genetic models are described which exploit the unique relationships that exist within the families of identical twins to obtain weighted least squares estimates of additive, dominance and epistatic components of genetic variance as well as estimates of the contributions of X-linked genes, maternal effects and three sources of environmental variation. Since all of the relationships required to achieve a resolution of these variance components are contained within each family unit, the model would appear to be superior to previous approaches to the analysis of quantitative traits in man.


[Reproducibility of emesis by orally administrated copper sulfate in cats]. Adult cats were administered oral threshold doses of copper sulfate every week. As the cats vomited in 70 out out 80 cases, the reproducibility was 88%. Peripheral vomiting threshold dose was 40 mg/head or less, while the threshold dose for oral copper sulfate emesis after T4 transection and vagotomy was more than 160 mg/head. The following method is thus proposed for application in evaluating an antimetic for oral copper sulfate. Adult cats are to be given the emetic once a week. The threshold dose should be determined in three dose levels; 10, 20, 40 mg/head. The cats with a threshold of more than 40 mg or latency of less than 5 min or of more than 45 min are to be excluded. Inhibition of emesis or a considerable prolongation of latency is the sign of an antimetic action. A positive action of an antiemetic must be followed by another test with the threshold dose of copper sulfate alone. If the cat does not respond to the threshold dose after 2 administrations, the case must be excluded. It is considered positive when 3 cases are inhibited among 4 or more than 50% among more than 5.


[Effectiveness of several antiemetics in vomiting induced by orally administrated copper sulfate in cats and dogs]. Long marketed antiemetics, which are still used in medicine or veterinary medicine and whose sites of actions were either assumed to be peripheral or unknown, were subjected to evaluation as antiemetics, against oral copper sulfate emesis. In cats, ethyl aminobenzoate 0.5 g/head, cerium oxalate 0.1 g/head, pinelliae tuber 0.5 g/head and gelatin 2.0 g/head were not effective. In dogs, ethyl aminobenzoate 0.5 g/head, pinelliae tuber 2.0 g/head and hange-syasintoo 2.0 g/head were not effective. Cerium oxalate 0.1 g/head was found to have a slight antiemetic action.


[Central effects of clonazepam].

[Central effects of clonazepam]. The effects of oral administration of clonazepam, a new benzodiazepine derivative (F. Hoffmann-La Roche), on the central nervous system were compared with those of diazepam and several anticonvulsants in mice and rats. 1) Clonazepam exhibited a moderate inhibitory effect on the locomotor activity observed with open-field situation in mice and no effect in rats, while it inhibited markedly the rearing behavior in both animals, the duration of action being approximately six hours. 2) Clonazepam potentiated the methamphetamine-induced hyper-locomotor activity in mice whereas trimethadione had no effect. 3) Clonazepam inhibited with a moderate potency the conditioned avoidance response and response to a fixed-ratio (FR 20) schedule of food reinforcement in rats, the potency being a little weaker than that of diazepam. 4) The muscle relaxant effect of clonazepam determined by the traction test was slightly more potent as compared with that of diazepam. Thiopental hypnosis was markedly potentiated after clonazepam. 5) The clonic (CL), tonic-flexor (TF) and extensor convulsions (TE) induced by pentetrazol were strongly inhibited after clonazepam in mice, anticonvulsant potency against CL and TE of clonazepam being approximately 23 and 21 times stronger than that of diazepam, 3333 and 3846 times that of trimethadione, and over 3047 and 178 times that of phenytoin, respectively. Clonazepam reduced markedly CL and TE elicited by bemegride with about 12 to 14 times stronger potency than diazepam. On the contrary, the anticonvulsant effect of clonazepam against TE of maximal electroshock seizure evoked by supramaximal current was weak, the potency being 0.71 times weaker than that of phenacemide, 0.14 times than phenytoin and 0.24 times than phenobarbital. By the combined administration of clonazepam with other anticonvulsants such as trimethadione and phenytoin against pentetrazol convulsion, and phenacemide, phenytoin and phenobarbital against maximal electroshock seizure, the antagonistic effect of these anticonvulsants was potentiated by 4 to 5 times. 6) The acute toxicity (LD50) of clonazepam was weak but that of phenacemide or phenytoin was potentiated to a certain degree by combined administration with clonazepam. The results suggest that clonazepam has a psychopharmacological profile similar to that of benzodiazepines with a particularly potent anticonvulsant effect on pentetrazol and bemegride convulsions, and the anticonvulsant effect is synergic with that of other anticonvulsants.


[Effect of ursodesoxycholic acid and cholic acid on intestinal absorption of cholesterol]. A study was carried out to investigate the effect of ursodesoxycholic acid and cholic acid on intestinal absorption of cholesterol-4-14C administered p.o. to lymph-fistula rats. The results indicated that within 24 hours, the labeled cholesterol detected in thoracic duct lymph of the control group was 21.0% of the administered cholesterol-4-14C, whereas the group treated with cholic acid (250 mg/kg p.o.) was found to have an increased value of 30.6% absorbed cholesterol. In comparison, the group treated with the same dose of ursodesoxycholic acid showed a decreased value of 12.1%, indicating an inhibitory effect on cholesterol absorption by the compound. This differential effect of the two compounds was also observed in a time-course study.


[Conditioning of emotional behavior originating at the hypothalamus. (2) Effects of drugs on conflict-induced behavior models]. We attempted induction of the conflict behavior by the locus pattern, thumping and conditioning under hypothalamic stimulation in rabbits. Flight was classified as continuous and non-continuous. Thumping was elicited post stimulus. Conditioning responses were obtained during the autonomic-somatic responses, continuous run and non-continuous run. All drugs were administrated orally. Dosage of each drug used in this experiment was calculated by determining the dosage which caused the same degree of muscle relaxation and sedative action in mice (see, Fig. 1) or general behavior in rabbits. Diazepam 5 mg/kg, chlordiazepoxide 10 mg/kg and nitrazepam 1 mg/kg caused non-continuous flight in the smooth run, but chlorpromazine 5 mg/kg induced continuous run with snarling. Methamphethamine 5 mg/kg induced continuous run and autonomic-somatic tonus pattern, and phenobarbital-Na 30 mg/kg depressed the animal to the stage of flight. Diazepam, chlordiazepoxide and phenobarbital-Na inhibited thumping, while chlorpromazine, nitrazepam and methamphethamine increased the incidence of thumping. Chlorpromazine transformed the non-continuous run into autonomic-somatic responses of conditioning. Diazepam transformed the non-continuous run into a continuous one. Phenobarbital-Na transformed the non-continuous run into autonomic-somatic responses. Methamphethamine transformed the non-continuous run into a withdrawal response. It would thus appear that the non-continuous flight, thumping and the non-continuous run conditioning are forms of conflict in behavior which can be applied to study of emotional response with psychotropic drugs.


[Inhibition of harmaline induced tremor by L-threo-3, 4-dihydroxyphenylserine, an L-norepinephrine precursor]. Effect of 3, 4-dihydroxyphenylserine (DOPS), a norepinephrine precurosr, on harmaline tremor was investigated in mice to elucidate the role of norepinephrine in the genesis of tremor. 1) Spontaneous motor activity was inhibited by L-threo-DOPS (200 mg/kg i.p.). 2) Tremor induced by harmaline (5 and 7 mg/kg i.p.) was enhanced by alpha-methyl-p-tyrosone (200 mg/kg i.p.). 3) The development and duration of tremor induced by harmaline (10 mg/kg i.p.) were inhibited significantly in a dose dependent manner by L-threo-DOPS (50, 70, 100, 150 and 200 mg/kg i.p.), but neither by D-threo-DOPS (200 mg/kg i.p.) nor DL-erythro-DOPS (200 mg/kg i.p.). 4) L-threo-DOPS (200 mg/kg i.-.) had no effect on the development of tremor induced by tremorine (5 and 10 mg/kg i.p.), while lacrimation and diarrhea caused by tremorine was markedly inhibited. 5) Administration of harmaline (10 mg/kg i.p.) produced an increase in brain 5-hydroxytryptamine content but not in that of norepinephrine. Administration of L-threo-DOPS (100 mg/kg i.p.) increased the norepinephrine content but not the 5-hydroxytryptamine content in the brain. Inhibition of harmaline tremor induced by L-threo-DOPS is attributed to the L-norepinephrine converted from L-threo-DOPS and the involvement of a noradrenergic mechanism in harmaline tremor has to be considered.


[Catecholamine metabolism. Physiopathology and pathobiochemistry]. Metabolic studies on isolated rat organs showed that vanillyl mandelic acid (VMA) is dehydrogenated by liver homogenate and suspension of pseudomonas fluorescens. In the 100000 g supernatant fraction of homogenated rat kidneys enzymatic activity was sufficient for kinetic studies and the examination of substrate specificity, pH dependence and changes with age and sex of rats. Norepinephrine secretion and resorption may be influenced by antiarrhythmic drugs in vivo: after lidocaine, there was no change of the tyramine dependend norepinephrine depletion of rat hearts, Verapamil inhibited the uptake of norepinephrine by rat hearts. Immobilisation of the animals led to a significant decrease in the myocardial levels for norepinephrine and epinephrine, which was antagonised by recreation of 24 hours. Adumbran prevented the catecholamine depletion, so that the influence of the limbic system may be important for the mediation of sympathetic stimuli. Emotional stress was shown to lead to an elevated plasma level of norepinephrine, cortisole and free fatty acids. In patients undergoing surgery, the application of Valium and Fortral inhibited these stress reactions. After physical stress, which did not lead to significant changes of the catecholamine level in plasma of healthy volunteers, there was an elevated norepinephrine plasma level in patients with cardiac failure, which could be prevented by different antiarrhythmic drugs. The excretion of norepinephrine and VMA of these patients was also increased. It was shown, that sinus rate rose according to norepinephrine levels in plasma in this group, which was contrary to the behaviour of healthy volunteers, showing neither a norepinephrine overflow nor increased heart rate. In patients with atrial fibrillation, AV-conduction also increased with elevated norepinephrine levels in plasma. There was, however, no direct correlation. Determination of the cardiac AV-difference of plasma-norepinephrine levels showed a cardiac catecholamine overflow, which could be inhibited by specific and nonspecific antiarrhythmic drugs (Verapamil, Practolol). In studies of catecholamine and glucocorticoid metabolism, no significant difference of catecholamine and VMA could be determined in patients with normal blood pressure and arterial hypertension. There was, however, a tendency to increased values in patients of the last group. In postural hypotension, plasma levels and urinary excretion of catecholamines were decreased and showed no variation after physical stress in orthostasis and after nicotine. In patients with tumors of the adrenal cortex and medulla, a close relation of the disturbance in catecholamine and corticoid metabolism was revealed.


[The role of ketone bodies in nonshivering thermogenesis in cold-adapted rats (author's transl)]. In spite of the ill fame of useless products of metabolism that only accumulate in the blood under abnormal conditions, ketone bodies (KB) have now regained the fame as an important fuel of respiration for the brain and muscles in prolonged starvation in humans and other animals. In cold-adapted animals, ketosis has been well documented, but little is known about the physiological role of KB in cold adaptation. In this study, role of KB in nonshivering thermogenesis was investigated in warm- and cold-adapted rats with or without norepinephrine (NE) loads, a decisive lipid metabolizing hormone in cold adaptation. First, levels of blood KB and plasma free fatty acid (FFA) and FFA turnover rate were examined along with rectal temperature under continuous infusions of various doses of NE. Good dose-response relationship of rectal temperature was obtained with increasing doses of NE in cold-adapted rats, but not in warm-adapted ones. The levels of blood KB and plasma FFA in cold-adapted rats remained almost unaltered during the observation period of 30 minutes. Infusion of NE, however, greatly enhanced FFA turnover rate in cold-adapted rats as compared with controls. Next, fractional turnover rate and half-life of beta-OH-butyrate were obtained from decay curves of endogenous beta-OH-butyrate, using functionally eviscerated rats in which no KB were produced. Calculated turnover rate of KB was also found significantly increased in cold-adapted rats. Finally, the effect of NE on the production of KB was examined using the liver in situ perfusion technique of Mortimore with minor modifications. Viability of the liver preparation was guaranteed by sufficiently high rates of bile production as well as oxygen consumption. The production rate of KB in the preparation under palmitate loads of physiological medium concentration was significantly higher in cold-adapted rats than in warm-adapted controls. However, no significant effect of NE loaded simultaneously was observed in either group. Quantitative analysis shows that the turnover rate of KB in vivo essentially equals the production rate in the perfused liver when no exogenous NE is added. In contrast NE infusion the turnover rate in vivo was almost doubled that of the perfused liver. All these results indicate KB are used as an important energy source with the significancy of FFA in nonshivering thermogenesis. Moreover, it may be surmised that production of KB in vivo, particulary in the cold-adapted state, is effected by other unknown factors than NE.


[Copper metabolism under the disturbed circulation of the liver (Eck's fistula) in dogs. The relation between copper content and histologic changes in the brain (author's transl)]. The relation between copper content and histologic changes in the brain under disturbed copper metabolism was studied. In this experiment, three groups of mongrel dogs were used, i.e. (1) Copper administration only, (2) Copper administration with Eck's fistula, (3) Normal control. The copper content in each brain tissue was measured by the atomic absorption spectrophotometry and routine histologic preparations with some special stains were examined. Following results were obtained. (1) In the group of longstanding copper administration, there were no remarkable histologic changes in the brain, and the significant increase of copper content was not demonstrated in the cortex, medulla and basal ganglia. (2) The group of copper administration with Eck's fistula was divided into two subgroups by the presence or absence of icterus. (a) Without Icterus: Diffuse gliosis and type II Alzheimer's neuroglia were observed and copper content in the brain increased up to 1.8 fold of that of normal control. (b) With Icterus: The histologic changes which were observed in the non-icteric group were more severe especially in the brain stem containing vacuolar degeneration, shrinkage and ghost appearance of the nerve cells, hemorrhages and demyelination, but there were no marked histologic changes in cortex and medulla. The copper content of the brian increased up to 2 fold or more of that of normal control. Copper was not demonstrated histochemically in all groups. The copper content in the cortex and basal ganglia was almost the same but that of medulla was half as much as that of the formers in every group of this experiment. The difference of the copper content between cortex which had no remarkable histologic changes, and basal ganglia which had aforementioned severe changes, was not proved, and also the difference between the groups with icterus and without icterus was not demonstrated. The copper content did not always correlate with the severity of the histologic changes. From these results, it can be suggested that copper does not play a significant role in pathogenesis of specific histologic changes of the brain.


Renal proliferative arteriopathies and associated glomerular changes: a light and electron microscopic study. This presentation reports the light and electron microscopic findings relating to the vascular and glomerular changes in the kidney in a series of 25 patients having malignant hypertension, the hemolytic-uremic syndrome, scleroderma, or toxemia of pregnancy. The pathologic changes were generally similar in each of the diseases studied, the changes being related more to the severity and duration of injury than to the specific disease. Vascular narrowing was due mainly to intimal thickening, and by light microscopy the lesions were categorized as onionskin, mucinous, or fibrous with or without associated elastosis. Intimal erythrocyte extravasation, fibrinoid necrosis, and luminal thrombosis were also seen. Electron microscopy provided additional morphologic information: Myointimal cells were found to be the cellular component in each type of intimal thickening; it was possible to distinguish collagen from large intimal accumulations of basement membrane material; mucinous intimal material was characterized ultrastructurally; and fibrinoid necrosis was identified as a lesion inconstantly associated with cellular necrosis and consisting mainly of fibrinoid material and small deposits of fibrin. It seems likely that there is a common pathogenesis for intimal thickening in a variety of diseases and that this involves endothelial cell damage and increased permeability, leakage of serum and erythrocytes into the intima, and a healing reaction of the vessel wall was developing from migration of smooth muscle cells into the intima with subsequent myointimal cell proliferation and fibrogenesis. A common glomerular change in all diseases studied was a striking accumulation of electron lucent material between the endothelium and the lamina densa of the basement membrane. This lesion was interpreted as a manifestation of acute ischemia.


Primary gastric choriocarcinoma: case report with review of world literature. A case is herin reported of a primary gastric choriocarcinoma occurring in an elderly woman associated with very high titers of circulating chorionic gonadotropin. Unlike the majority of previously reported cases, in which adenocarcinomatous components were present, the tumor in this case was purely trophoblastic. The clinicopathologic findings of the previously reported cases are summarized, along with the various theories of histogenesis.


Familial occurrence of gonadal tumors in XY females with breast development. Three sisters with XY gonadal dysgenesis are presented. All of them have primary amenorrhea and breast development, and in two, a gonadal tumor, seminoma (dysgerminoma) was found. The danger of gonadal neoplasm in XY females with normal female build is emphasized.


Spermatogenesis in an infertile XYY man. Testicular histology and meiosis has been studied in an XYY male patient identified at an infertility clinic. This man was found to have an XYY set chromosome complement in 15% of spermatogonial metaphases. There was no evidence of 2 Y chromosomes at diakinesis but there appeared to be a slight excess of sperm with a fluorescent Y body.


A case of true hermaphroditism with 45X/46XY mosaicism. The authors describe a case of true hermaphroditism of mainly female phenotype, ambiguous genitalia, and ovotestis. The cytogenetic revealed 45X/46XY mosaicism and an absence of Barr bodies.


Conditions affecting primary cell cultures of functional adult rat hepatocytes. 1. The effect of insulin. The conditions for obtaining representative, primary adult rat hepatocyte cultures were explored. The methods applied included enzymatic liver perfusion which was nondestructive to hepatocytes, the prevention of aggregation of dissociated cells and the selective attachment of viable cells. These procedures yielded a recovery of 50% of the liver cells which gave rise to cultures representing 14% of the total liver cells. The cultures were composed of homogeneous epithelial-like cells cytologically similar to hepatocytes and possessed a number of liver-specific enzymes. There was virtually no cell division initially and most cells died between 24 and 48 hr. Insulin enhanced the attachment of the liver cells, altered their morphology, but did not prolong cell survival.


Further studies on the intracellular behavior of Torulopsis glabrata. The growth of Torulopsis glabrata was inhibited in glucose-peptone broth containing 10 to 20% normal human serum. Addition of iron to the medium diminshed the fungistatic effect. The intracellular growth of T. glabrata was remarkably restricted within mouse macrophages maintained in vitro, but this growth restriction was not caused by the limitation of iron imposed by the serum in the medium. The intracellular growth of T. glabrata within a very small percentage of the macrophages was not obviously related to the failure of lysosomal fusion to the phagosomes in those cells. The studies did not permit definite conclusions regarding the viability of the inhibited yeasts, but results suggested that a large portion of them survived. Potentially misleading artifacts of the technique for assessment of the intracellular behavior of the fungus were detected and are discussed.


Uptake of diphtheria toxin and its fragment A moiety by mammalian cells in culture. Evidence suggesting that diphtheria toxin reaches the cytoplasm of susceptible mammalian cells by two independent mechanisms is presented. A schematic model describing the two processes of toxin entry into the cell is developed. One process of toxin uptake considered to by physiologically significant is passage of the protein toxin through the plasma membrane. This most likely happens by binding of fragment B to receptors on the membrane and by subsequent toxin-membrane interaction so that ultimately fragment A, the enzymatically active moiety, is transported tothe cell interior. This process, which ultimately leads to cessation of protein synthesis and cell death, involves a comparatively small number of toxin molecules. A second mechanism of toxin uptake is by classical pinocytosis. The majority of toxin taken into the cell is accomplished by this process. The fate of toxin taken into HEp-2 cells via pinocytosis is proteolysis by lysosomal enzymes. Thus, such vesicle-bound toxin is ordinarily not expressed biologically. Evidence suggesting that ammonium chloride provides total protection to diphtheria toxin-susceptible cells by preventing entry of toxin by the specific receptor-associated process is also provided; data showing that the ammonium salt immobilizes bound toxin on the plasma membrane of HEp-2 cells are presented. Finally, it is suggested that actively endocytic cells such as guinea pig macrophages interact with toxin in a significantly different manner than do nonphagocytic cells.


Levels of prostaglandins in the small intestine of rats during primary and secondary infection with Nippostrongylus brasiliensis. The level of PGE is increased 10-fold in intestinal tissues during primary infection of the rat with Nippostrongylus brasiliensis. Peak levels (ca. 7,000 pg/cm) were assayed in the jejunal site of infection on day 7 of infection and similar levels were recorded in 'post-infection' (ileal) segments at this time. The level of PGE in 'post-infection' segments showed further increase to 12,000 pg/cm on day 10. The level of PGE also increased in 'pre-infection' (duodenal) but this was delayed by 4-5 days. The level of PGF also increased during primary infection (from about 100 to 950 pg/cm) but this occurred after expulsion. Increase in the level of PGE occurred earlier (at 3-4 days) during secondary challenge given 19 days after primary infection, but the PGE levels followed the primary response when challenge was given 10 weeks after primary infection. It is suggested that PGE plays a dual role in parasite immunity. (1) PGE may directly affect metabolism of the parasite. In this event it is also suggested that protective antibodies cause the release of PG. (2) Elevated levels of PGE act indirectly by affecting gastrointestinal function which alters the microenvironment at the site of infection. The duodenal migration of parasites may be due to this effect of PGE.


Stress incontinence of urine.

Stress incontinence of urine. Urethrocystographic, clinical and surgical aspects have been studied in 50 premenopausal women with Green's type II stress incontinence. Thirty patients were surgically treated by a retropublic (Marshall-Marchetti-Krantz) procedure and 20 by a vaginal (Kelly-Kennedy) method. The radiology and surgical procedures were performed by the author, always using the same sequence and standardization. The relative position of the vesical angle and the urethral inclination were studied. Twelve months after operation the retropublic technic showed topographic modifications that were statistically more significant that those for the vaginal procedure. No difference was noticed in clinical results for the Kelly-Kennedy and Marshall-MArchetti-Krantz procedures (85% and 80% of safety respectively).


Some comments on the urethral syndrome.

Some comments on the urethral syndrome. In three-quarters of the patients with urethral syndrome, chronic urethritis was found endoscopically. Of the remainder some precentage could be said to have a psychosomatic disorder. Therefore, the patients with unexplained urethral syndrome were unexpectedly few. Although urethral syndrome attacks adult females of all ages, the incidence was highest in the middle age range. Against this, chronic bacterial cystitis was spread evenly over the entire age range. The urethra was rather narrow in women with urethritis, but of normal size in those without urethritis, even if they had urethral syndrome. A satisfactory result was obtained through the topical administration of steroid solution, even where general treatments, including the use of antibiotics, have failed.


An experimental implantable urinary sphincter. An artificial urethral sphincter was implanted in 10 dogs and observed for periods up to 18 months. No necrosis or strictures of the urethra developed. The plastic part of the device failed in 10 to 18 months.


Electron microscopic localization of lactate dehydrogenase in osteoclasts of chick embryo tibia. Lactate dehydrogenase (LDH) was localized in osteoclasts of fixed and unfixed 19-day chick embryo tibias using a copper ferrocyanide capture reaction and osmiophilic polymer generation. This study revealed that: (I) LDH activity in fixed, briefly rinsed osteoclasts was associated principally with limiting membranes of cytoplasmic vacuoles and vesicles and with the plasma membrane; (2) LDH activity in unfixed osteoclasts was associated only with mitochondria; and (3) some mitochondria were stained in fixed tissue given a long rinse. These results indicate that: cytoplasmic LDH diffused out of unfixed tissue; mitochondrial LDH was inactivated by formaldehyde in fixed tissue; and formaldehyde-inhibited mitochondrial LDH can be reactivated by a long rinse. Although the vesicles that stained for LDH activity were found in all parts of the cell, they were concentrated near the ruffled border, and there is evidence that they contained material from the bone surface. These results suggest that the LDH associated with cytoplasmic vesicles of the osteoclasts may be important in processing of material resorbed from the bone surface and that osteoclasts mitochondria may utilize lactate from the bone fluid for energy production.


[Diagnosis and treatment of conductive deafness due to middle-ear effusion in childhood (author's transl)]. Results of follow-up examinations of 132 ears with serous or mucous middle ear effusions indicate that this disease is common in childhood and is also found in adults. In childhood adenoid growth as well as repeated catarrhal infection of the paranasal sinuses and the weakness of the levator palati play an important role in the development of middle ear effusions and deafness. With the aid of the operation microscope we were able to recognize this characteristic disease through the tympanic membrane. We perform regularly adenoidectomy together with paracentesis in this condition. In cases which had undergon several adenoidectomies we limit our treatment in the first instance to paracentesis. If cure is not so achieved we insert an inert tube into the tympanic membrane after the paracentesis. In persistent and difficult cases of glue ear radiation of the eustachion tube and antrotomy may be necessary.


Failure of colostral immunoglobulin transfer in calves dying from infectious disease. Serum IgG1 concentrations of calves less than 3 weeks old and dying from infectious disease were significantly lower (P less than 0.01) than those of clinically normal calves. Fifty percent of the dead calves had serum IgG1 concentrations that were more than 2 standard deviations below the normal mean, and an additional 35% had IgG1 concentrations that were more than 1 standard deviation below the normal mean. Low IgG1 concentrations were attributed to failures in passive transfer of colostral immunoglobulin. The few calves dying of noninfectious causes generally had normal serum immunoglobulin concentrations. The results of this study emphasize the importance of adequate colostral intake and absorption to the neonatal calf. In view of the large numbers of calves that die from neonatal infection each year, it may be assumed that failure in passive transfer, as reflected by low serum immunoglobulin concentrations, is one of the most important factors influencing neonatal calf mortality.


Rapid digestion and flameless atomic absorption spectroscopy of mercury in fish. A rapid method is described for the determination of total mercury in fish samples. The sample is digested with nitric acid-sulfuric acid-potassium permanganate, and then reduced and aerated for measurement by flameless atomic absorption spectrophotometry. The average recoveries of organic and inorganic mercury added to fish were 93 and 95%, respectively. The uniformity of mercury levels in shark tissue has also been investigated.


Identification of cholesterol as a possible interference in the analysis of fish tissue for industrial chemical residues. The presence of cholesterol in cleaned up fish tissue extracts has been established by gas-liquid chromatography and confirmed by mass spectrometry. It was found both in extracts cleaned up by gel permeation chromatography and in those extracts prepared by the AOAC method for nonfatty foods. It is not normally observed in routine residue analyses because selective gas chromatographic detectors such as electron capture are commonly used.


Thin layer chromatographic determination of sterigmatocystin in cereal grains and soybeans. A method has been developed for determining sterigmatocystin in yellow and white corn, barley, oats, rye, sorghum, brown and wild rice, and soybeans. A partition column packed with activated magnesium silicate was used for cleanup. Averaged recoveries are 104% in white corn, 114% in rye, 100% in oats, 134% in brown rice, 96% in barley, 105% in sorghum and wild rice, and 92% in soybeans. The limit of sensitivity is 50 mug/kg for any of these commodities.


Distribution of the low molecular weight form of eukaryotic elongation factor 1 in various tissues. Since it was shown that the low molecular weight form of the eukaryotic polypeptide chain elongation factor 1 (EF-1alpha) is highly unstable but could be stabilized by the addition of 25% (v/v) glycerol to all the buffer solutions (Nagata, S., Iwasaki, K., & Kaziro, Y. (1976) Arch. Biochem, Biophys, 172, 168-177), its distribution in several tissues has been investigated under conditions where it is stabilized. The results obtained indicate that EF-1alpha is present as a predominant species in all the tissues examined, such as pig liver, rat liver, rabbit reticulocytes, and Artemia salina cysts. Furthermore, it is suggested that the high molecular weight form of the elongation factor 1 contains a new elongation factor, EF-1beta, in addition to EF-1alpha, while the low molecular weight form contains little EF-1beta.


Kinetic competence of a phosphoryl enzyme intermediate in the glucose-1,6-p2 synthase-catalyzed reaction. Purification, properties, and kinetic studies. Glucose-1,6-P2 synthase of beef brain which catalyzes the formation of glucose-1,6-P2 and glycerate-3-P from glycerate-1,3-P2 and glucose-1-P has been purified 700-fold with an overall recovery of 19%. The purification procedure involves an ammonium sulfate fractionation of the crude extract, DE52 and hydroxylapatite column chromatography and isoelectric focusing. The isolated enzyme appears to be homogeneous by sodium dodecyl sulfate gel electrophoresis. Its molecular weight is estimated to be about 70,000 by gel filtration on Sephadex G-200 which agrees with the value obtained by sodium dodecyl sulfate gel electrophoresis. A phosphoryl enzyme intermediate in the catalytic reaction is indicated by the following evidence: glycerate-1,3-P2[1-32P] labels the enzyme. The label is removed by acceptor substrates such as glucose-1-P. Using a rapid quenching device at 23 degrees and pH 8.0, the first order rate constant for phosphorylation of the enzyme is 20 s-1, compared with an overall rate with the best acceptor, glucose-1-P, of 19 s-1. Dephosphorylation by glucose-1-P is at 37 s-1. Mg2+ is required for both phosphoryl transfers and the overall reaction. In the complete reaction the fraction of enzyme that is phosphorylated depends on the concentrations of glycerate-1,3-P2 and the concentration and nature of the acceptor in a way that could be predicted from the steady state parameters, the Km values, and the kinetic constants observed for the single turnover. Reciprocal plots of initial rates as a function of both substrate concentrations are families of parallel lines. The 32P-labeled phosphoryl enzyme intermediate was found to be acid-stable and somewhat alkaline-labile. Phosphoserine was identified from the partial acid hydrolysate of a protease digest of [32P] phosphoryl enzyme by two-dimensional thin layer chromatography.


Glucose metabolism of mammalian erythrocytes. Under physiological conditions, erythrocytes of the horse metabolized 638 +/- 37 (+/-SE) nmoles glucose/ml cells/hr at 37 degrees C compared to 942 +/- 31 for the cat, 1,329 +/- 44 for the dog, and 1,485 +/- 43 for man. On an absolute basis, pentose phosphate metabolism was similar between species, with species differences in erythrocyte glucose tulization attributable to differences in Embden-Meyerhof pathway metabolism. By examining pentose phosphate pathway recycling, it appears that some functional compartmentation exists within erythrocytes.