Biological controls in mixed trichostrongyle infections. The parasite reaction phenotype of 80 lambs was determined with natural infection on pasture. These lambs were then assigned to outcome groups and randomized into treatment groups. Developing some simple but unusual analytical techniques it was possible to demonstrate that mixed natural infections of trichostrongyles with the chief biomass component Haemonchus contortus cayugensis protected against superinfections with 35000 infective larvae but protected against superinfections with 1000 infective larvae per day only within the limits characteristic of the enclosing barn ecosystem. The limits of the barn ecosystem were less constraining on more diathetic sheep than the natural pasture infection. Internal evidence from the experiment suggests that the interaction of sheep X pasture X infective trichostrongylid larvae constrains the parasite biomass in the host in more diathetic animals at significantly lower levels. Anaphylactoid 'self-cure' did not occur in this experiment but something like premunition certainly did.


[Nitrogen metabolism of broilers fed 15-N labeled wheat.5. Skin, feathers and 15-N balance]. In the present study 17 four-week-old broilers received a ration supplemented with 15N labelled wheat for a period of 2 days. The 15N excess was 295mg. 3 birds each were killed 3, 6, 12, 60 and 108 hours after administration of the last 15N dose. The present communication provides data on the N and 15N content of the bone skin, feathers and data on the 15N balance. On an average, 11% of the amount of 15N administered were found in medullar bones. A fairly uniform pattern of 15N labelling was observed at the different test points of the skin while the time pattern of 15N frequencies in the feathers was non-directional. 15N' balance showed that at all the 6 test point the range of error for the recovery rates was less than 3%. The present study substantiated the suitability of the stable N isotope for use in N metabolism trials.


Active immunization against tetanus of children suffering with diabetes mellitus. The authors report on the anti-tatanus reimmunization of 76 diabetic children and adolescents hospitalized in the Diabetic Ward for Children in Varshetz. All patients had insulin-deficient diabetes and they were treatedwith insulin Lente. The diet was limited and physiological. Thirty-n-ne of the patients had diabetes in a state of compensation, while the remaining 37 patients were in a state of decompensation without acetonuria. The reimmunization was well tolerated and no strong general and local reactions were observed. The authors do not establish statistically significant differences during an overnight period in the blood sugar level and the degree of glucosuria before and after reimmunization. They conclude that the immunization does not disturb the metabolic processes. On the 20th day after the reimmunization 98.6% of the children showed a high antitoxic titre, which suggests that the capacity of diabetics to elaborate a good anti-tetanus immunity is unimpaired. It is recommended that patients with diabetes should be immunized against tetanus during a stage of stabilized diabetes metabolism.


Renal localization of 67Ga-citrate in renal amyloidosis: case reports. In scans taken 72 hr after intravenous administration of 5 mCi of 67Ga-citrate, both kidneys were clearly visible in two cases of histologically proven renal amyloidosis. Neither patient had clinical manifestations or laboratory evidence of concurrent inflammatory disease or tumor involving the kidneys. Increased renal concentration of lysosomal organelles and increased affinity of 67Ga for amyloid material contained in the organelles could explain the renal uptake of 67Ga in amyloidosis.


Diagnosis and epidural hematoma by brain scan and perfusion study: case report. By using the arterial and venous phases of an anterior cerebral perfusion study, which showed downward displacement of the sagittal sinus, and the finding of a "rim" on the delayed scans, the specific diagnosis of epidural hematoma was established.


[Transfusion of blood with HL-A type. A study with 2,380 donors (author's transl)]. Patients receiving repeated transfusions sooner or later develop an anti-HL-A immunization. To prevent or to delay this immunization by transfusing "deleucocyted" blood appears in the long run, to be a vague hope. So, it became essential to use blood from donors typed for the HL-A system. In this study we have examined what would happen if a pool of 2,380 HL-A type donors were available for 110 recipients. Twenty five per cent of the recipients will have blood available from more than 5 donors with the same HL-A types as themselves, and 71% of them can receive blood from more than 5 donors sharing at least one antigen at each locus (and with no antigen known to be different between them).


Factor structure of locus of control in children. The purpose of the present study was to identify the factor structure of the Children's Nowicki-Strickland Internal-External Control scale. To this end, the responses of Ss at the elementary (n = 449), junior high (n = 388) and high school (n = 389) were subjected to factor analysis procedures. Both general and specific factors were found. It was suggested that locus of control scales be prepared so as to yield a general as well as specific, score to allow the investigator to choose the appropriate measure of locus of control for his purposes.


Low frequency dynamic viscoelastic properties of human mitral valve tissue. The dynamic viscoelasticity of the anterior leaflet of human mitral valves was investigated by subjecting the ventricular aspect of membranous samples of the tissue to sinusoidal fluid pressures. The frequency range of the stressing function used was from 0.5 Hz to 5 Hz. The storage modulus of the tissue was found to be independent of the stressing frequency and an average value of 2.55 (SE = 0-12) X 10(8) dyn cm-2 was observed. Losses were small and the phase shift between stressing function and resulting strain and hence the loss modulus increased with frequency. For the frequency range investigated the phase shift varied from 0.04 to 0.06 rad and the loss modulus was of the order of 10(7) dyn cm-2. From the observed data it was concluded that any tissue substitute used in mitral valve replacement should be rather inextensible and have a low loss modulus. Also under normal physiological conditions, the mitral valve cannot bulge into the left atrium during peak ventricular systole and hence events in the cardiac cycle, such as the presence of the atrial pressure 'c' wave, that involve the distensibility of the valve need to be re-examined.


Use of levamisole in the treatment of parasitic tracheobronchitis in the dog. Of 36 dogs with a history of chronic coughing or dysponea examined by bronchoscopy, eight were found to have parasitic tracheobronchitis. Oral treatment with the anthelmintic levamisole at a rate of 7.5 mg per kg bodyweight was found to be successful in every case.


Seasonal changes in the plasma retinol-binding holoprotein concentration of sheep. 1. Seasonal variations in plasma retinol-binding holoprotein concentration have been found in two groups of entire ewes and in a control group of wethers (castrated rams). 2. In each group retinol-binding holoprotein level was minimal in summer and increased to peak values in September just before the autumn breeding season.


Complexity and specificity of polysomal poly(A+) RNA in mouse tissues. Base sequence complexities of polysomal poly(A+)RNA from mouse embryo, brain, and liver have been estimated by hybridization to homologous cDNA to be approximately 7 x 109, 1.5 x 1010, and 7 x109 daltons, respectively. By annealing each cDNA with a large excess of total mouse embryo DNA, the genes coding for the polysomal poly(A+) sequences were shown to be unique. Heterologous hybridization experiments showed that the high abundance class of poly(A+) sequences in one tissue is not identical with the high abundance class in other tissues. However, at least 55%, and possibly more, of the poly(A+) RNA in one tissue is present in the poly(A+) RNA of another tissue.


Crystalline 3-phospho-d-glycerate kinase from horse muscle. Phosphoglycerate kinase has been isolated in crystalline form from horse muscle. A convenient isolation procedure is described that yields homogeneous enzyme of specific activity 700 units/mg (30 degrees C). The enzyme is monomeric, and has a molecular weight 47 000. Of the eight cysteine residues in the protein, two react rapidly with Nbs21 with the concomitant loss of the catalytic activity. Since the isolation of phosphoglycerate kinase from yeast (Bücher, 1955) there have been several reports of purification methods yielding enzyme approaching molecular homogeneity, from rabbit muscle (Beisenherz et al., 1953; Czok and Bücher, 1960; Rao and Oesper, 1961; Avramov and Repin, 1965; and Scopes, 1969) and from chicken muscle (Gosselin-Rey, 1965). Crystalline material has been isolated from human erythrocytes (Hashimoto and Yoshikawa, 1962), and from yeast and rabbit muscle (Krietsch and Bücher, 1970). Cystallographic work on phosphoglycerate kinase from horse muscle by Blake et al. (1972) and Blake and Evans (1974) has prompted mechanistic interest in the enzyme, and we report here a simplified isolation procedure and some properties of the crystalline material from this source.


Inactivation of bovine liver 2-keto-4-hydroxyglutarate aldolase by cyanide in the presence of aldehydes. Kinetic data show that the irreversible inactivation of liver 2-keto-4-hydroxyglutarate aldolase observed when the enzyme is incubated with an aldehydic substrate (or substrate analogue) in the presence of cyanide is a biphasic process and can, under certain conditions, involve a direct interaction between the enzyme and cyanide. The kinetic data are consistent with a scheme consisting of three competing reactions: (1) irreversible addition of cyanide to the enzyme-substrate Schiff base intermediate, (2) reversible cyanohydrin formation between cyanide and the aldehydic substrate (or substrate analogue), and (3) an interaction of cyanide with the enzyme which is not substrate dependent. Approximately 0.4 mol of cyanide is associated with 1 mol (120 000 g) of enzyme when 2-keto-4-hydroxyglutarate aldolase is incubated with [14-C]-cyanide followed by exhaustive dialysis; an ionic attachment possibly at a carboxylate binding site, is suggested. Whereas native enzyme, not treated with cyanide, has ten Nbs2-titratable sulfhydryl groups, approximately one less such group reacts with Nbs2 when the aldolase is incubated with cyanide (in the absence of aldehydic substrate). It is suggested that the binding of cyanide results in a conformational change of the enzyme; conformational changes in the presence of cyanide are confirmed by circular dichroism spectra.


Biosynthesis of prostaglandin F2alpha from arachidonic acid and prostaglandin endoperoxides in the uterus. Formation of prostaglandin F2Alpha in the cow and guinea pig uterus microsomes was studied using 14C-labeled arachidonic acid and prostaglandin H2. The total conversion of arachidonic acid was of a low order and underwent fluctuations during the estrous cycle of the guinea pig, being highest towards the end of the cycle. Injections of beta-estradiol-3-benzoate also resulted in higher activity of the uterine prostaglandin synthetase. The uterine prostaglandin synthesizing system appeared to differ in several respects from that present in seminal vesicles, with regard to the proportions of the products formed and the effects of various agents, e.g. reduced glutathione. An inhibiting factor which supressed the fatty acid cyclo-oxygenase was found to be present in uterine preparations. Prostaglandin endoperoxide (prostaglandin H2) was very efficiently reduced to prostaglandin F2alpha by cow and guinea-pig uterus microsomes. Prostaglandin G2 also gave rise to prostaglandin F2alpha. Prostaglandin E2, on the other hand, was not reduced. Both the inhibiting factor and the endoperoxide reducing activity are likely to be parts of a highly specialized mechanism that modulates prostaglandin F2alpha formation in the uterus.


A high-resolution NMR study (1H, 13C, 31P) of the interaction of paramagnetic ions with phospholipids in aqueous dispersions. 1H-, 13C- and 31P-NMR spectra of egg-yolk phosphatidylcholine (PC), phosphatidylserine (PS) and phosphatidic acid (PA) and cosonicated mixtures of these phospholipids were obtained from ultrasonicated dispersions containing Pr3+, Eu3+, Gd3+ and Mn2+ ions. The differences in chemical shift values, deltaN, between the "inner" and "outer" resonance signals for the different nuclei of the polar head group of egg-yolk phosphatidyl choline provide information about the average distances of the paramagnetic ion within the polar groups of the phospholipid molecules. In the Pr(2H2O)3+n/egg-yolk phosphatidylcholine system the ions are nearest to the phosphate and -CH2CH2 group, respectively but relatively far from the N(CH3)3 group of the polar head group of the lipid. The integral analysis of the 1H-NMR spectra obtained from dispersions containing Pr3+ and Mn2+ ions enables us to calculate the number of the polar groups in both sides of the egg-yolk phosphatidylcholine bilayer, the size of lipid vesicle and to give some features of the arrangement of the phospholipid molecules in cosonicated egg-yolk phosphatidylcholine/phosphatidylserine vesicles. At p2H 8.3 in PC/PS mixtures an extreme asymmetry is observed with PS preferentially in the outer side of the membrane. This side contains approximately three times more PS than PC molecules. Some comments are made concerning the quantitative integral analysis of proton-noise decoupled 31P-NMR spectra as obtained from similar phospholipid mixtures by Michaelson et al. and Berden et al.


[Role of the adenyl cyclase system in achieving the immunogenesis-stimulating action of bacterial lipopolysaccharides--pyrogenal and endogenous serum pyrogen]. Experiments were conducted on rabbits immunized intraperitoneally with corpuscular typhoid vaccine; the number of antibody-forming cells in the spleen proved to increase after tha administration of bacterial lipopolysaccharide (LPS)--pyropeneal, and endogenous serum pyrogen (EPS) together with theopylline. The data obtained indicated that the adenylcyclase system played a certain role in the mechanism of the stimulating action of LPS and EPS.


[Correction of the immune response to sheep erythrocytes in mice of different strains using polyelectrolytes]. Inbred mice--C57BL, A, C57BR, C3H and CBA strains with a high, medium and low response to sheep red blood cells (SRBC) were injected with polymeres--poly-4-vinylpyridine (P-4-VP) and polyacrilic acid (PAA), and were immunized with SRBC; production of antibody-forming cells in their spleen was determined. It was shown that the administration of P-4-VP to C57BL mice was followed by a 5--fold increase of the immune response and injection of PAA--by a 4-fold increase. Injection of P-4-VP and PAA to CBA mice increase the immune response 2.5- and 2.8-fold, respectively. In the C57BR mice P-4-VP increased the immune response 4 times, and PAA--4.7 times. Treatment of C3H mice with P-4VP was followed by a 2-fold and with PAA by a 2.4-fold increase of the immune response. Injection of P-4-VP and PAA to CBA  mice had no effect on the immune response. Thus, with the increase of the immunological reactivity to sheep erythrocytes in the mentioned strains of mice there was observed a reduction of the enhancing effect of the polymeres to the immune response.


Factor XI activity and factor XI antigen in homozygous and heterozygous factor XI deficiency. A relatively potent antiserum against highly purified, unactivated human factor XI antigen was raised in a rabbit. This antiserum, after concentration, neutralized 50% of the factor XI clotting activity of a standard normal plasma at an antiserum dilution of 1/900. The antiserum was used in a neutralization-inhibition assay to study the relation between factor XI clotting activity and factor XI antigen in plasma from ten unrelated patients with homozygous factor XI deficiency and from 12 heterozygous family members of these patients. No evidence of factor XI antigen significantly in excess of factor XI activity was found in either group. All data to date have been consistent with the hypothesis that hereditary factor XI deficiency represents a genetic disorder resulting from the absence of factor XI molecule. Severity of bleeding in factor XI deficiency could not be correlated with the level of factor XI activity or factor XI antigen.


Diencephalic dopamine receptors and regulation of pituitary-testicular system. Injection of dopamine, DOPA or apomorphine into the third brain ventricle resulted in an increase in the plasma testosterone level. Pimozide, blocking dopamine receptors, as well as complete, long-posterior and posterior but not anterior deafferentation of the MBH abolished the stimulating action of dopamine. After direct injection of dopamine into the arcuate nucleus there was no stimulating effect on the pituitary-gonadal system. At the same time, local injections of dopamine and apomorphine into the medial part of the subthalamus or periventricular gray matter of the caudal thalamus were followed by a distinct increase in the plasms testosterone level. The conclusion is drawn that the brain dopaminergic neurons activating the pituitary-gonadal system in the male are localized outside the hypophyseotrophic area of hypothalamus in zona incerta and caudal part of the thalamus. After stimulation of diencephalic dopamine receptors, nervous impulses are transmitted to the MBH through the posterior hypothalamic pathways.


Free-operant avoidance behavior in rats with lateral septal lesions: Effect of shock intensity. The purpose of experiment 1 was to examine the relationship between shock intensity and normal rats' free-operant (Sidman) avoidance performance on a 3-component, multiple schedule. The results showed an inverted U-shaped relation between response rate and shock intensity, but no consistent relation between shock rate and shock intensity was found. Experiment 2 showed that lateral sptal lesions produced a bidirectional change in reactivity to electric shock. An increased reactivity was observed in the initial sessions starting on the 11th day after the surgery, while a reduced reactivity was observed in the final sessions when stable performance had been reacquired. These changes were not a function of the waning of general hyperreactivity with post-operative recovery: the septal lesions in this study did not produce any "sham rage". From 40 to 80% of the variance in response rates was accounted for by this bidirectional change in the reactivity to shock.


Interaction between radiation and drug damage in mammalian cells. II. The effect of actinomycin-D on the repair of the sublethal radiation damage in plateau phase cells. The effect of actinomycin-D (AMD) on radiation damage repair was studied in plateau phase V79 Chinese hamster cells. Sublethal radiation damage repair, as demonstrated by survival fluctuations following two x-ray exposures separted by time, was observed in our plateau phase cells. Plateau phase cells exposed to 0.01-0.04 mug/ml AMD (a nontoxic regimen to 8 hours) between x-ray exposures were less able to repair sublethal damage. If plateau phase cells were plated at low dilutions into fresh medium (conditions for resuming exponential growth) immediately after the first x-ray dose, and exposed to 0.01--0.04 mug/ml AMD until the second dose, inhibition of sublethal damage repair and additional cell killing were observed particularly at 0.04 mug/ml AMD. It is suggested that radiation-drug damage interactions should be studied in plateau phase cells and in cells resuming exponential growth after plateau phase (possibly analogous to "recruitment"), as well as in exponential phase cultures.


Immunodiagnosis and monitoring of gonadotrophin-producing metastases in the central nervous system. Measurements of human chorionic gonadotrophin (HCG) concentration in plasma and cerebrospinal fluid (CSF) have been made on patients with gonadotrophin-producing tumors. In the absence of brain metastases the spinal fluid concentration is, within wide limits, proportional to that in the plasma. In 73 patients with gestational choriocarcinoma the mean plasma/spinal fluid ratio was 286 with a lowest value of 64. In five patients with gonadotrophin-producing teratomas the mean ratio was 208 with a lowest value of 104. In 29/33 patients with brain metastases the plasma:CSF ratio was less than 60 at the time confirmatory evidence of brain metastases was obtained. One patient with a gonadotrophin-producing teratoma had a brain metastasis that apparently failed to produce HCG and this metastasis failed to show the histologic features of choriocarcinoma. Monitoring the plasma:CSF ratio provided evidence of brain metastases in 13/18 patients who were undergoing chemotherapy for extensive metastatic disease before confirmatory evidence was obtained by other methods. The lead-in time between assay diagnosis and diagnosis by other methods ranged from 1-20 weeks. Monitoring the plasma:CSF ratio provided a means of observing the response of cerebral metastases to therapy. In some patients the CSF HCG concentration exceeded the plasma concentration indicating that the higher CSF values in patients with CNS metastases cannot be attributed to impairment of the blood-brain barrier. Direct secretion of tumor products into CSF or indirect secretion into CSF via cerebral extracellular fluid evidently occurs. In contrast with radiographic and radionucleide detection methods, a chemical marker indicates the metabolic activity of tumor cells within the central nervous system and provides a basis for monitoring that activity.


Immunochemical detection of human enzymes in hybrid cells. Rabbit antisera have been produced against each of three purified human enzymes: a cytoplasmic form of NADP-linked isocitrate dehydrogenase (IDH, EC 1.1.1.42), phosphoglucose isomerase (PGI, EC 5.3.1.9), and hypoxanthine guanine phosphoribosyltransferase (HGPRT, EC 2.4.2.8), and they have been used for immunoprecipitation reactions to detect human-specific enzymes in various human-mouse somatic cell hybrids. Under optimal conditions, enzyme activity was eliminated from human cell lysate, but no reduction of enzyme activity was found in the mouse cell lysate. Differential enzyme precipitation by these human-specific antisera was observed in human-mouse hybrid cells. Analysis on starch gel electrophoresis revealed that not only the human homodimer, but also human-mouse heterodimer molecules, in cases of PGI and IDH, were precipitated. Thus this method is sensitive and allows quantitative determination of human-specific enzymes. The presence of a human-specific enzyme identified by this method correlated with the presence of a particular human chromosome permitting assignments of the human cytoplasmic forms of NADP-linked IDH, human PGI, and human HGPRT genes to chromosomes 2, 19, and X, respectively. These assignments are consistent with published data (Ruddle, 1973).


Partial purification of the ovalbumin gene. Cellulose-bound DNA complementary to ovalbumin mRNA was used in a continous hybridization system to isolate single-stranded DNA molecules containing the ovalbumin gene. Fragmented DNA segments containing the ovalbumin gene were enriched 300-350 fold in one cycle of purification. Two cycles of purification resulted in a DNA fraction which was enriched 2300 fold in the ovalbumin sequence. The method is suitable for purification of the ovalbumin sequence from both sheared DNA fragments, as well as larger molecular weight DNA containing more than twice the number of nucleotides necessary to code for ovalbumin mRNA. The chromatographic procedures were specific and reproducible. In addition, the recovery of ovalbumin DNA was essentially quantitative (80-100%), even when large amounts of starting DNA (70-75 mg) were used. This purification scheme should also be useful for the enrichment of other unique sequence gened form eucaryotic DNA.


Pseudoejection sound in hypertrophic subaortic stenosis: an echocardiographic correlative study. Phonoechocardiographic studies were performed in 23 patients with hypertrophic subaortic stenosis. In ten patients a distinct systolic sound was recorded, usually along the lower left sternal border. In eight of these cases the sound was low or medium frequency. Unlike the ejection click of valvular aortic stenosis, the sound began 40-100 msec after the upstroke of the indirect carotid pulse and occurred close to the initial peak of the carotid pulse. This sound, termed pseudoejection sound, was associated with systolic anterior movement (SAM) of the anterior mitral leaflet on the echocardiogram. In all six patients studied with simultaneous phonoechocardiograms, the pseudoejection sound coincided with the sudden halting of SAM of the anterior mitral leaflet. Following provocative maneuvers the sound became louder, and its timing, as well as the sharp halting of SAM of the mitral leaflet, occurred earlier in systole. The pseudoejection sound probably results either from impact of the anterior mitral leaflet against the interventricular septum or more likely from deceleration of blood flow in the left ventricular outflow tract. The echo data support the association of the pseudoejection sound with significant left aventricular outflow obstruction,


Use of a cholesterol-rich bovine lipoprotein to enhance cholesterol concentrations in the preparation of serum control materials. We describe a process for preparing a cholesterol-rich lipoprotein extract from bovine serum. The material is used to enhance cholestrol concentration up to 3.5 g/liter in a clear control material from human serum. The optical clarity of the enriched serum is only slightly affected by lyophilization and reconstitution. The lipoprotein additive neither produces any evidnet interference in the measurement of the commonly analyzed consitutents nor affects the stability of components after lyophilization and reconstitution.


Pigeon breeders disease. III. A study of a family exposed to doves. A family of five individuals exposed to doves in their home were studied to evaluate possible factors leading to sympotmatic hypersensitivity pneumonitis documented in one member of the family, while the other exposed members remained asymptomatic. Antbodies against pigeon serum were domenstrated by radioimmunoassay in all subjects, and significant lymphocyte reactivity against pigeon antigens was demonstrable in three of the five members. The presence of serum antibody activity or lymphocyte reactivity against avian antigens or both did not differentiate the sick from the well family subjects. However, the possibility that increased exposure to antigen may correlate with higher degrees of antibody and lymphocyte reactivity as well as clinical disease is suggested from observation made in this family. No correlation was found between HL-A type and either symptomatic disease or immunological reactivity in these subjects.


In vitro sensitivity of enteric bacteria to epicillin, chloramphenicol, ampicillin and furazolidone. Strains of Salmonella typhi (148) and Salmonella paratyphi A (27) isolated from the blood of patients with clinical features of enteric fever were tested in vitro for their sensitivity to epicillin, ampicillin, chloramphenicol and furazolidone. Results from both the disc and tube dilution methods showed that greater percentages of the two strains were sensitive to epicillin than to the other antibiotics.


Ultrastructure of cytoplasmic nucleolus-like bodies and nuclear RNP particles in late prophase of tipulid spermatocytes. Late prophase stages of Pales ferruginea (Tipulidae) spermatocytes were examined by means of conventional electron microscopic section technique, combined with cytochemical methods. The cytoplasm of cells in diakinesis contains nucleolus-like bodies (NLB) 1 mum in diameter which are formed in diplotene at the pores of the nuclear membrane. They are compound structures consisting of fibro-granular RNP material which is associated wth one or two electron-dense gobules. The RNP material has a hollow core which contains polyribosomes. The NLBs possibly indicate rRNA gene amplification. At diakinesis the nucleus contains numerous electron-dense RNP particles scattered throughout the chromatin-free karyoplasm, and associated with the condensed chromosomes. The diameter of the chromation associated particles is markedly higher (mean 630 A) than that of the free particles (mean 540 A). The RNP particles seem to be aggregates of 200 A subunits. They are regarded as transcription products of chromosomal genes.


Studies on the role and mode of operation of the very-lysine-rich histones in eukaryote chromatin. Nuclear-magnetic-resonance studies on nucleoprotein and histone phi 1-DNA complexes from marine invertebrate sperm. Proton magnetic resonance and other measurements have been carried out in order to study the behaviour of the lysine-rich histones phi 1 in the sperm chromatin of certain marine invertebrates. Well defined particles (12 S) have been obtained from this chromatin by nuclease treatment. Chromatin solubility as a function of ionic strength shows a relaxation at salt concentrations higher than in the case of calf thymus nucleoprotein. Nuclear magnetic resonance (NMR) studies show that the release of histone from DNA occurs both in chromatin and in the reconstituted complexes at practically the same ionic strength as solubility relaxation. The higher the arginine content of a given phi 1, the higher the ionic strength at which both effects take place. The NMR results demonstrate that arginine residues are bound more strongly than lysine residues. The data overall show that phi 1 histones play a role in the contraction mechanism of sperm chromatin similar to that of H 1 histone in calf thymus chromatin. The highly contracted state of sperm chromatin is directly related to the increased arginine content of the phi 1 histone.


Deleterious effects of copulation in Drosophila females as a function of growth temperature of both sexes. In Drosophila females, copulation always results in a reduction of longevity but the gravity of the phenomenon varies according to growth temperature. The harmful effect of adult males, or aggressiveness, is maximum when growth took place at a middle temperature. Female sensitivity, on the other hand, is at a maximum when larvae were reared at exteme, low or high, temperatures.


Characteristics of the nucleolini observed under the electron microscope. Observations with the electron microscope permitted us to ascertain that in molluscs and echnioderms oocytes and in malignat tumour cells, the nucleolini, already seen with the phonton microscope, correspond to the "clear fibrillar zones/. These present fibrils 40-60 A thick, spread throughout a very clear matrix. All around these zones there are other closely thickened and interlacing fibrils.


Effect of delta-THC on ethanol withdrawal in mice. Delta-THC (10 mg/kg, i.p.) administered to mice immediately after withdrawal from a 3-day exposure to ethanol vapor was found to intensify withdrawal reactions. No effect was seen when delta-THC was administered chronically during the exposure to ethanol.


Large translocation t(3q-;4p+) as probable cause for semisterility. A couple with one phenotypically normal son and two recorded abortions complained of apparent inability to have more children. The father, and also his son, proved to be carriers of the translocation t(3q-;4p+). Almost certainly, the length of the translocated segment, practically the whole 3q, would cause either translocation chromosome by itself to lead to prenatal death. Furthermore, the fertility of the carrier father may be considerably decreased if in a sufficient number of spermatocytes the meiotic orientation of the chain or, less probably, the ring formed by 3, 3q-,4p+, and 4 is such that the two 3-centromeres go to one pole and the two 4-centromeres to the other, not to mention an even more abnormal 1:3 segregation.


Atropine-induced inhibition of the enhanced CCK release observed in alcoholic dogs. As it has been previously shown, the pancreatic secretory response to an intraduodenal infusion of oleic acid is increased in animals accustomed to daily ethanol consumption compared to matched controls. This action has been verified in dogs provided with a Thomas cannula and consuming 2 g kg-1 ethanol or not, daily since 3 years. An intravenous infusion of 0.75 mg kg-1 h-1 of atropine suppresses the difference between alcoholic and non-alcoholic animals. Therefore, the increased release of CCK-PZ in response to meal, which is characteristic of chronic alcoholic animals, is under cholinergic control.


Chronic gastric ulcer in the rat produced by wounding at the fundo-antral junction. Standardized 8-mm mucosal wounds were made in the rat stomach at the rumeno-fundic junction, fundus, fundo-antral junction, and antrum, and examined at intervals up to 8 months. All wounds at the rumeno-fundic junction and fundus were healed within 4 weeks and at the antrum within 12 weeks. The majority of wounds at the fundo-antral junction remained chronic and at 8 months 41 of 50 rats still had ulcers 2 mm or greater in diameter. These findings suggest that the fundo-antral junction is an area which is susceptible to the development of chronic ulcers. This is a simple and reliable method of producing chronic gastric ulcers and does not alter the continuity or disturb the function of the gastrointestinal tract.


The effect of metiamide, an H2-receptor antagonist, in the prevention of experimental stress ulcers. Because of evidence that Metiamide, an H2-receptor antagonist, strongly inhibits gastric acid secretion, the present study was designed to test the hypothesis that Metiamide will prevent bile salt-induced stress ulcers during hemorrhagic shock. Forty dogs were bled and maintained for 4 1/2 hr at a mean blood pressure of 40 to 50 mm Hg. In group A, 10 dogs received 300 mg of Metiamide orally 45 min before bleeding and 10 dogs received normal saline. The pylorus was occluded before bleeding and 100 ml of 15 mM bile salt were instilled into the stomach and aspirated at the end of 4 1/2 hr. At the time the animal was killed after 48 hr, no ulcers were seen in the stomachs of dogs treated with Metiamide. Sixty per cent of the dogs in the untreated group developed multiple gross ulcers. In group B the effect of Metiamide on the disappearance rate of H+ ion was measured by instillation of 50 mM HCl + 15 mM bile acid. No difference was noted in the rate of H+ ion disappearance between Metiamide-treated and control dogs. Also, in 5 normotensive dogs the rate of H+ ion disappearance was measured before and after treatment with Metiamide, and the loss of H+ was identical for both periods. Metiamide was effective in preventing stress ulcer in this experimental model. The protective effect of Metiamide is probably due to its inhibitory effect of H+ ion secretion.


Factors in the etiology of restraint erosions in parabiotic rats. Rats joined in surgical parabiosis for 25 to 30 days were tested by restraining one member of the pair on a movable cart while allowing the second member to remain free to move about. These rats were compared with other rats joined in pseudoparabiosis for 25 to 30 days so that the behavioral aspects of parabiosis were reproduced but blood sharing did not occur. Pseudoparabiotic rats were also tested on the movable cart at the end of the recovery and adaptation period. Gastric erosions formed in the unrestrained members of both the parabiotic and pseudoparabiotic pairs with an incidence and severity that did not differ statistically. The experimental results fail to support the hypothesis that humoral factor(s) are critical in restraint ulcer pathogenesis in the rat.


Rostellar development in the larva of Multiceps endothoracicus (Kirschenblat, 1948). The bladder of the larval M. endothoracicus bears several anlages which bulge in the shape of cellular buds in to the bladder cavity. A canal is differentiated from the thickened tegument which invaginates into these buds. The position of the tegument covering the bottom of the cellular anlage is more or less horizontal. The subtegumental layer is distinct. The rostellar cone differentiates from an increased number of cells situated below the tegument which covers the bottom of the canal. The tegument of the cone is covered with fine hooklets which break off gradually into the lumen of the canal. After the division of the cone into a bulb and praebulb we observed that the latter becomes overgrown by a lateral extension of the adjoining canal wall). Larger hooklets with a differently staining base were found in the thickened fold separating the praebulb from the bulb. The blade of the definitive hook differentiates from the modified tegument of this area, and from the larger hooklets after the formation of the hook organ. The lumen of the canal, and later also the spiral canal, harbour leukocytes from the tissue of the host; these attack the superficial microtrix border seen with the electron microscope. The tegument of the canal consists of a microtrix border (depth 6--7 mum), of a thick zone of distal cytoplasm with oval and rod-shaped electron dense bodies. The cytoplasm traverses the basal fibrillar membrane between the sub tegumental muscles and joins the subtegumental cells.


Cytological and chemical changes in cell walls of Rhodotorula gracilis. III. Characteristics and life cycle of the yeast. The phenotype and genotype of six strains of the genus Rhodotorula Harrison and of one strain of the genus Cryptococcus Phaff et Fell, with anomalous thickening of cell walls were investigated. The present studies showed that the strains investigated represent different stages of the life cycles of the genus Rhodosporidium Banno. The anomalous thickening of the cell walls can be explained by extreme conditions resulting in the formation of surviving forms (teliospores, chlamydospores).


[Forensic significance of the fate of ingested ethanol (author's transl)]. Distribution and disappearance of ethylalcohol (ethanol) were studied using either human blood in vitro or blood sampling or corpse blood after premortal ethanol ingestion. 1) Ethanol is distributed among plasma and blood cells according to a partition law. The partition coefficient was found to vary depending upon ethanol concentration and corpse phenomenon post mortum. 2) Ratio of plasma ethanol concentration to blood cells increases below 0.5 mg/ml and decreases above 0.5 mg/ml of ethanol concentration in the whole blood. The ratio was found to be more smaller in the corpse blood. The concentration ratio in blood cells to whole blood varies in a similar fashion. 3) Accuracy of determination of ethanol levels in the corpse whole blood was not always satisfactory, partly because distribution of ethanol in the blood varies postmortum and partly because ratio of plasma to blood cells also varies. Some attempts were made using statistical manipulations of hematocrit values or ratios of plasma to whole blood, to correct the errors without recommendable success. 4) After deliberabe evaluation of errors, possibly incurred when the value post mortum is used instead of the premortal values, it is concluded that the postmortal value can be used as an indicator of premortal ebriety within the reasonable limit of errors. 5) No changes in the distribution of ethanol were observed within 24 hours when the blood is kept in ice box immediately after sampling, and also in the blood ethanol concentration for at least 20 days under similar conditions. In contrast, blood samples, if left at room temperature, deteriorate rapidly within 5 days.


[Studies on esterification of fatty acids in rat liver mitochondria (author's transl)]. Esterification of fatty acids in rat liver mitochondria was studied following portal or intraperitoneal administration of various 14C-fatty acids. Incorporation of the labelled fatty acid was most marked into phospholipids (especially, phosphatidylcholine and phosphatidylethanolamine) and triglycerides in intact mitochondria and the inner mitochondrial membranes. In contrast, in the outer membranes the injected fatty acids remained free without esterification. It is concluded that esterification of fatty acids occurs in the inner membranes of rat liver mitochondria.


[Chronological changes in leukemias in Hokkaido (author's transl)]. The various leukemias in Hokkaido for the past 20 years are analysed in its change of the mortality rate, the type of leukemia, and the clinical picture. The observation years are divided into 4 periods: I (1951-1957), II (1958-1962), III (1963-1967), IV (1968-1972). 1. The adjusted mortality rate of leukemia shows a rising tendency through these periods: I (1.7), II (3.0), III (3.5), IV (3.6). 2. The adjusted age-specific mortality rate of leukemia in period II, III, and IV, show a remarkable increase in all age brackets compared with period I, especially in older age brackets. The rate of the "60-69 years old" bracket gradually rises as I (3.1), II (4.8), III (5.9), IV (7.2). The rate of the "over 70 years old" bracket is I (1.3), II (1.8), III (5.0), IV (9.3). It is characteristic that these brackets has the highest mortality rate in period IV. 3. Some comparisons between period I and IV are described as follows: (1). Monocytic leukemia had decreased from 10.6% (I) to 1.4% (IV) and chronic myelogenous leukemia has slightly increased in period IV. (2). In acute type, the cases of aleukemic leukemia tends to increase in period IV and its initial symptoms changes into mild gradually. (3). In chronic type in period IV, splenomagaly is common in the initial symptom and while blood cell counts in these cases are above 10 x 10(4)/cmm.


Effect of long-chain acyl-CoA on the omega-oxidation of monocarboxylic acid. In vitro addition of stearoyl-CoA in 10,000 x g supernatant of rat livers, first enhanced the rate of omega-oxidation of free [1-14C] stearate, until a concentration of the acyl-CoA reached to the same level of that of the substrate. Further addition of stearoly-CoA decreased gradually the oxidation rate.


[A case of the A. Vertebralis sinistra direct arising from the arcus aorta (author's transl)]. A case of the left vertebral artery direct arising from the Arcus aorta, falling under C-type in ADACHI (1928), was encountered among the corpses dissected for practice in Hokkaido University, school of medicine. The Arcus aorta has four branches; Truncus brachiocephalicus, A. carotis communis sinistra, A. vertebralis sinistra, and A. subclavia sinistra. The A.vertebralis sinistra has passed through at first into Foramen transvwrsarium of the vertebra cervicales quintus (C5). The length of this artery between the origin of the Arcus aorta and the fifth cervicales was 120 mm. The other measurements of arteries were shown in table 1.


Effect of testosterone on long-term organ cultures of canine prostate. Organ cultures of rodent and human prostate glands have shown marked differences in their morphological response to testosterone. In this study, explants from 19 canine prostate glands were cultivated for a minimum of 9 days in Trowell's T-8 medium. Groups of explants were exposed to media containing from 0.05 to 100 mum testosterone. While the higher testosterone levels (50 and 100 mum) markedly decreased explant viability, explants cultivated at lower levels (0.05 to 5 mum) appeared similar to control explants in testosterone-free Trowell's T-8 medium. Atmospheric mixtures containing either 95% or 50% oxygen were equally effective. Shortly after the cultures were initiated, large amounts of secretory product were liberated into the lumen. After 9 or more days in vitro, glandular epithelium appeared cuboidal and never revealed the acid phosphatase-rich secretory granules seen in the preculture control. However, the epithelium exhibited an increase in alkaline phosphatase and lipid content following cultivation.


Interaction of Mycoplasma dispar and Mycoplasma agalactiae subsp. bovis with bovine alveolar macrophages and bovine lacteal polymorphonuclear leukocytes. Mycoplasma dispar and Mycoplasma agalactiae subsp. bovis survived or grew in cultures of bovine lacteal polymorphonuclear leukocytes or bovine alveolar macrophages. In the presence of specific bovine antibody, macrophages and polymorphonuclear leukocytes appeared to kill both species of mycoplasma. Specific rabbit antisera also promoted the killing of these mycoplasmas by bovine macrophages but had no demonstrable activity for bovine polymorphonuclear leukocytes. It is suggested that phagocytosis of these mycoplasmas by bovine cells occurs only in the presence of specific antibody. The experiments also indicate that differences exist between bovine polymorphonuclear leukocytes and macrophages with regard to their receptor sites for immunoglobulins.


Interaction of virulent Mycoplasma pneumoniae with hamster tracheal organ cultures. Exposure of hamster tracheal organ cultures to virulent Mycoplasma pneumoniae leads to alterations in ribonucleic acid (RNA) and protein biosynthesis and metabolism of the respiratory epithelium. An examination of the turnover rates of RNA and protein in infected tracheal organ cultures indicates that the rates of degradation of both prelabeled host cell RNA and protein are similar to those of uninfected controls. Infected tracheal organ cultures shifted to a nonpermissive medium within 24 h after infection and further incubated in the nonpermissive medium for 72 or 96 h behaved as normal uninfected cultures in terms of metabolic precursor uptake. Under these conditions, mycoplasmas remained attached to the respiratory epithelium. Cell membranes prepared from virulent mycoplasmas by several procedures neither attached to nor altered the metabolic activity of tracheal cultures. These data indicate that the intimate contact between virulent mycoplasmas and the respiratory epithelium does not alone account for the subsequent interruption of host cell metabolism but must be accompanied by continued multiplication and biochemical function of attached mycoplasmas.


Adhesive properties of Vibrio cholerae: adhesion to isolated rabbit brush border membranes and hemagglutinating activity. Adhesion of vibrios to the small intestine may occur (i) by association of the bacteria with secreted mucus gel or (ii) by adherence of the bacteria to the surface of epithelial cells. In the present study, vibrios readily adhered to isolated brush border membranes obtained from rabbit intestinal epithelial cells. Adhesion was temperature dependent and required the presence of divalent cations such as calcium. The agglutination of human O erythrocytes by Vibrio cholerae was observed also, and the hemagglutination test appeared to detect the same mechanism that was involved in the adhesion of vibrios to brush borders. When the bacteria were grown in broth they were adhesive and hemagglutinating, but vibrios grown on agar plates or suspended in buffer for 15 min at 37 C lacked these abilities, even though they retained undiminished motility. These two model systems differed, however, in that strontium promoted only adhesion to brush borders. The significance of this difference remains to be determined. Vibrios were observed to penetrate intestinal mucus gel and occasionally to become entrapped in it. However, there was no evidence that vibrios attached to mucus gel.


Adhesive properties of Vibrio cholerae: nature of the interaction with isolated rabbit brush border membranes and human erythrocytes. Nonmotile vibrio mutants lacked the ability to adhere to rabbit intestinal brush border membranes and to agglutinate human group O erythrocytes, but motile revertant vibrios isolated from such strains expressed adhesiveness equivalent to that of the original parent. Two possible explanations for the relation between vibrio motility and adhesion in these assays systems are (i) that the rate of adhesion depends on the rate of chance contact brought about by motility, and (ii) that the flagellum either acts as a carrier for the bacterial adhesin or may itself be the adhesin. The present study indicates, however, that the lack of adhesion by nonmotile vibrios did not depend on motility as such and did not involve greater rates of elution. Increasing the rate of contact between nonmotile vibrio mutants and brush border membranes by compaction did not restore the adhesive properties of the defective strains. Accordingly, we speculate that the flagellum may function in some indirect way that allows the expression of the adhesive properties, such as by acting as a carrier for a specific vibrio adhesin. Adhesion to brush borders and agglutination of human group O erythrocytes was specifically inhibited by L-fucose and various glycosides of L-fucose and to a lesser extent by D-mannose. Vibrios adhered specifically to agarose beads that carried covalently linked L-fucose on their surfaces. The results suggest that L-fucose-containing structures of eukaryotic cell surfaces may function as receptors for the vibrio adhesin and may therefore be an important determinant of host susceptibility.


Immediate toxicity of high multiplicities of Chlamydia psittaci for mouse fibroblasts (L cells). One hour after suspensions of mouse fibroblasts (L cells) were exposed to 500 to 1,000 L-cell 50% infectious doses of Chlamydia psittaci (6BC), the L cells failed to attach to and spread out on solid substrates, phagocytosed polystyrene latex spheres at reduced rates, incorporated less [14C]isoleucine into protein, and had smaller soluble pools of nucleoside triphosphates. The infected L cells began to die at 8 h and were all dead by 20 h. Lower multiplicities of infection took correspondingly longer to kill the L cells. These effects of high multiplicities of C. psittaci on L cells will be referred to collectively as immediate toxicity. Similar effects were obtained with other strains of C. psittaci and C. trachomatis and with other cell lines. Ultraviolet-inactivated C. psittaci retained the ability to cause immediate toxicity, but heat-inactivated chlamydiae did not. C.psittaci cells had to be ingested by L cells before they were immediately toxic but, once they were phagocytosed, they did not need to multiply or to synthesize macromolecules in order to cause immediate injury to their hosts. Immediate toxicity was not the result of depression of energy metabolism, changes in the levels of intracellular cyclic nucleotides, or release of hydrolases from lysosomes. It was suggested that a lesion is produced in the plasma membrane of the L cell every time it ingests a chlamydial cell, that each act of ingestion produces an independent lesion, and that their injurious effects are additive. Thus, the more ingestion lesions there are, the faster the host cell dies. It was also suggested that induced phagocytosis, inhibition of lysosomal fusion, and death of mice and of cells in culture may all depend on a single activity of C. psittaci.


Characteristics of streptolysin S hemolysis. The characteristics of hemolysis produced by streptolysin S (SLS) were investigated in rabbit erythrocytes. Treatment of erythrocytes with SLS at various temperatures prior to incubation at 37 C revealed an initial temperature-dependent interaction between toxin and the cells. No subsequent hemolysis occurred when erythrocytes were exposed to SLS at 0 to 10 C; exposure to toxin at temperatures above 10 C gradually increased the amount of hemolysis that occurred at 37 C. Very little binding of toxin to erythrocytes or their ghosts, as detected by a decrease of hemolytic activity from toxin preparations, could be demonstrated at any temperature. The release of hemoglobin after the temperature-dependent process occur at virtually the same rate at 0, 22, or 37 C. The loss of intracellular rubidium-86 (Rb) and hemoglobin from SLS-treated erythrocytes was studied. Rb+ release significantly preceded the escape of hemoglobin, suggesting that colloid-osmotic processes play a role in SLS hemolysis.


Induction of interferon in ovine and human lymphocyte cultures by mycoplasmas. Mycoplasma pneumoniae, Acholeplasma laidlawii, M. arthritidis, and M. pulmonis were shown to induce interferon in the lymphocyte fraction of ovine peripheral blood leukocytes, but not in the polymorphonuclear leukocyte fraction. Human peripheral blood lymphocytes produced significant levels of interferon in response to infection with M. pneumoniae and M. synoviae. The antiviral substance induced by the mycoplasmas in human lymphocytes was characterized as interferon by the usual criteria.


Topographical relationship between sucrase and leucine beta-naphthylamidase on the microvilli membrane of rabbit intestinal mucosal cells. The topographical relationship between sucrase [EC 3.2.1.26] and leucine beta-naphthylamidase (LNAase) on the microvilli membrane of rabbit small-intestinal mucosal cells was studied assuming that where enzymes with different antigenicities, A and B, are situated in close proximity on the surface of microvilli vesicles, the agglutination of vesicles by anti-A antibody is inhibited by the previous binding of monovalent fragments of anti-B antibody to enzyme B on the surface of vesicles. Like anti-sucrase antibody, anti-LNAase antibody quantitatively agglutinated microvilli vesicles. It inhibited the membrane-bound LNAase activity in the same manner as the detergent-solubilized activity. This inhibitory effect of anti-LNAase antibody was not interfered with by monovalent fragments of anti-sucrase antibody. However, the monovalent fragments inhibited vesicle agglutination by anti-LNAase antibody as well as by anti-sucrase antibody. These results indicate that LNAase is located on the outer surface of microvilli vesicles and suggest that LNAase and sucrase are situated in close proximity on the membrane surface of microvilli vesicles.


The subunit structure of myosin from skeletal muscle of the early chick embryo. The subunit composition and immunological properties of two types of myosins, the 3S and 6S myosin components, from skeletal muscle of early chick embryos were studied by SDS-acrylamide gel electrophoresis and immunodiffusion techniques. It was shown that the 6S myosin in the early embryonic stage was composed of two heavy chains and three kinds of light chains, as is well-known in the complete myosin molecule, having the same molecular weights and the same antigenicities as corresponding subunits of the myosin from adult chicken skeletal muscle. The heavy chain of 6S myosin was also reactive with the antibody against the heavy chain of cardiac myosin. The embryonic 3S myosin was shown to be composed of a heavy chain which was roughly the same in molecular weight but not the same in antigenicity as those of adult or embryonic 6S myosin. No light chains were detected either electrophoretically or immunologically in the 3S myosin component.


Relationship between the structures of S-acyl thiol compounds and their rates of hydrolysis by pancreatic lipase and hepatic carboxylic esterase. About 100 S-fatty acyl thiol compounds designed as substrates for pancreatic lipase [EC 3.1.1.3] were synthesized and tested for susceptibility to hydrolysis by hog pancreatic lipase and hog hepatic carboxylic esterase [EC 3.1.1.1] using 5,5'-dithiobis(2-nitrobenzoic acid) as a chromogenic reagent to determine the hydrolytic rates of their S-acyl bonds. In general, the hydrolytic rates of S-acyl bonds by the lipase were fast with thioglycerol type thiol moieties, slow with dithioethyleneglycol type or monothiol type, and negligible with thiopolyol type. As for the acyl moieties, the hydrolysis of the S-acyl bonds was fast with C3-5 acyl groups, followed by C6-8 acyl groups, and the rate decreased as the acyl chain length deviated from these values or branched. On the other hand, the hydrolysis of S-acyl bonds by the esterase occurred with all types of S-acyl esters except for esters of long S-acyl chains. Of all the compounds tested with the lipase, the rate of hydrolysis of S-acyl bond was maximum with 2,3-dimercaptopropan-1-ol tributyroate [I], high with 3-mercaptopropane-1,2-diol tributyroate [II], but negligible with the analogous compound, 1,3-dimercaptopropan-2-ol tributyroate. Compounds [I] and [II] may be practically useful as substrates for lipase assay in human serum samples pretreated with phenylmethylsulfonyl fluoride, a potent inhibitor against both serum arylesterase [EC 3.1.1.2] and hepatic esterase, which attack [I] and [II].


Chemical composition and properties of soybean beta-amylase. The molecular weight of soybean beta-amylase [EC 3.2.1.2] was determined to be 57,000 daltons by the sedimentation equilibrium method, and the enzyme was found to consist of 494 amino acid residues. No difference was found in molecular weight or composition between two components of beta-amylase separated by ion-exchange chromatography. The N-terminus of the enzyme was not detectable by the fluorodinitrobenzene method or phenylisothiocyanate method, and the C-terminus was determined to be alanine by the carboxypeptidase [EC 3.4.12.2] method. Five half-cystine residues were found in the form of cysteine; all the sulfhydryl groups could be titrated by p-chloromercuribenzoate after denaturation of the enzyme with guanidine hydrochloride, but only some in the native enzyme. The rates of mercaptide formation of these groups were dependent on pH and were different from each other, all being much lower than the rate for the free sulfhydryl group in mercaptoethanol. Differential titration experiments at different pH's and in the presence of maltose showed that mercaptide formation by only one sulfhydryl group caused loss of activity, and the reaction was accompanied by changes in the environment around aromatic side chains in the enzyme, which were detected by difference spectra and fluorescence emission spectra. These facts suggest that modification of the sulfhydryl groups causes a conformational change of the enzyme. Some preliminary crystallographic data for crystals formed at pH 4.0 were obtained, and inactivation by heavy metal salts was examined in relation to the preparation of isomorphous heavy atom derivatives.


Kinetics of conformational change of troponin C induced by calcium. The kinetics of conformational change of troponin C (TN-C) induced by binding and removal of calcium ions were studied by measuring the fluorescence of tyrosine by stopped-flow spectrofluorometry. When the concentration of free calcium ions in the solution [Ca2+] was increased rapidly from 4X10(-9)M to 1X10(-3)M at neutral pH, a first-order reaction with a rate constant of 13.7 sec-1, which was preceded by a much faster reaction, was observed. In contrast, when [Ca2+] was reduced from 2X10(-3)M to 4.4X10(-8)M, two first-order reactions with rate constants of 7.4 and 0.78 sec-1, preceded by a much faster reaction, were observed.


Phenylalanine ammonia-lyase. Induction and purification from yeast and clearance in mammals. Yeast phenylalanine ammonia-lyase (EC 4.3.1.5) catalyzes the deamination of L-phenylalanine to form trans-cinnamic acid and tyrosine to trans-coumaric acid. Maximal enzyme activity in Rhodotorula glutinis (2 units/g, wet weight, of yeast) was induced in late-log phase (12 to 14 hours) of growth in a culture medium containing 1.0% malt extract, 0.1% yeast extract, and 0.1% L-phenylalanine. A highly purified enzyme was obtained by fractionation with ammonium sulfate and sodium citrate followed by chromatography on DEAE-cellulose and Sephadex G-200. The active preparation yielded a major component on three different polyacrylamide gel electrophoretic systems. Antisera to phenylalanine ammonia-lyase was raised in rabbits and detected by double immunodiffusion. The antigen-antibody complex was enzymatically active in vitro. The biological half-life of the enzyme was approximately 21 hours in several mammalian species (mice without and with BW10232 adenocarcinoma and B16 melanoma, rats, and monkeys) after a single injection; however, upon repeated administration, phenylalanine ammonia-lyase had a much shorter biological half-life. The onset of rapid clearance occurred earlier in tumor-bearing than in nontumor-bearing mice indicating a direct or indirect influence by the tumor on the biological half-life of phenylalanine ammonia-lyase.


Comparison of candicidin, levorin and trichomycin by means of high-performance liquid chromatography. A high-performance liquid chromatographic method for the separation of individual components of polyene macrolide antibiotics has been developed. The three heptaene macrolide antibiotics, candicidin, levorin and trichomycin, have been investigated and compared. In all instances these compounds proved to be complex mixtures. It is demonstrated that candicidin and levorin are identical, whereas trichomycin differs in composition from these two substances.


Recurrent goiter, hyperthyroidism, galactorrhea and amenorrhea due to a thyrotropin and prolactin-producing pituitary tumor. A 22-year-old woman with recurrent goiter, hyperthyroidism, galactorrhea, and amenorrhea due to a pituitary tumor is described. She had been treated surgically twice for recurrent goiter with tracheal compression. Despite clinical signs of hyperthyroidism and slightly elevated plasma thyroid hormone levels (T4: 11 mug/dl; T3: 189 ng/dl), without thyroid hormone replacement therapy the basal TSH level was elevated up to 23 muU/ml and could not be suppressed by exogenous thyroid hormones: even when the serum thyroid hormone levels were raised into the thyrotoxic range (T4: 16.2 mug/dl T3: 392 ng/dl), the basal TSH fluctuated between 12 and 29 muU/ml. The basal PRL level was elevated up to 6000 muU/ml. The administration of TRH (200 mug iv) led only to small increments of TSH and PRL levels. Bromocriptin (5 mg p.o.) or l-dopa (0.5 g p.o.) suppressed TSH and PRL values significantly. After transsphenoidal hypophysectomy, TSH and PRL were below normal and the patient development panhypopituitarism. The adenoma showed two cell types which could be identified as lactotrophs and thyrotrophs by electronmicroscopy and immunofluorescence. From these data we conclude that the patient had a pituitary tumor with an overproduction of thyrotropin and prolactin.


Diural TSH variations in hypothyroidism. There is a circadian variation in serum TSH in euthyroid subjects. A similar diurnal variation has been demonstrated in patients with hypothyroidism. In the present study the 24-hour pattern of serum TSH was investigated in eight patients with hypothyroidism of varying severity and in five hypothyroid patients treated with thyroxine (T4). There was a circadian variation in serum TSH in patients with hypothyroidism of moderate degree, and in patients treated for severe hypothyrodism with thyroxine. The pattern was similar to that found in normal subjects, i.e., low TSH levels in the daytime and higher levels at night. In severely hypothyroid patients, no diurnal variation in serum TSH was observed. A practical consequence is that blood samples for TSH measurements in patients with moderately elevated TSH levels are best taken after 1100 h, when the low day levels are reached.


Microrespirometer chamber for determinations of viability in cell and organ cultures. The effects of chemical, physical, and infectious cytotoxic agents on primary and cultured cells were evaluated by measurements of oxygen uptake for various time periods. A newly developed respirometer used a Clark oxygen electrode in a 1.0-ml chamber, with provisions for constant mixing and for temperature control of both the sample and electrode chambers. The device was unique because the electrode and instrumentation were provided by a clinical blood-gas analyzer. Oxygen uptake by blank controls was negligible, whereas cells and tissue consumed oxygen at rates of approximately 1 to 5 mul/h in a dose- and temperature-dependent fashion. Cyanide, heat, and freeze-thaw lysis reduced the oxygen uptake to less than 0.6 mul/mg per h. Infection of trachea organ cultures with Mycoplasma pneumoniae significantly reduced relative ciliary activity, tetrazolium reduction capacity, and oxygen consumption in a coordinated fashion.


Renal handling of Zn-alpha2-glycoprotein as compared with that of albumin and the retinol-binding protein. An unusual electrophoretic pattern of the urine from a patient with malignant lymphoma was observed. One of the major proteins, identified Zn-alpha2-glycoprotein (Zn-alpha2), was isolated from the urine and partly characterized. The Stokes radius was found to be 3.24 nm and the molecular weight, determined by sodium dodecyl sulfate polyacrylamide electrophoresis, 42,000. The plasma level in healthy individuals was 39 +/- 7 (SD) mg/liter. In 12 of 25 healthy individuals, Zn-alpha2 was measurable in the urine and was found to be 1.0 +/- 1.1 mg/liter. In 23 patients with chronic glomerulonephritis (CGN), in 9 with proximal tubular dysfunction (PTD), in 23 with various renal diseases (VRD), and in 10 with malignant lymphoma, the plasma level and the urinary excretion were compared with those of albumin (mol wt 67,000) and of the retinol-binding protein (RBP, mol wt 21,000). A close correlation was found between the urine-to-plasma (U/P) ratios of Zn-alpha2 and albumin in the patients with CGN, whereas in the PTD patients the U/P ratios of Zn-alpha2 and RBP were correlated. No significant renal arteriovenous difference in Zn-alpha2 could be demonstrated. The Zn-alpha2 excretion was increased also in two patients with malignant lymphoma and proteinuria of a tubular pattern. The plasma Zn-alpha2 varied inversely with the glomerular filtration rate in the patients with renal disease, but was normal in those with malignant lymphoma. The results are consistent with the assumption of a sieving coefficient of Zn-alpha2, substantially exceeding that of albumin, but notably lower than that of smaller low-molecular-weight proteins. An increased excretion of Zn-alpha2 may be due to increased glomerular permeability as well as to defective proximal tubular reabsorption.


Effect of 1-deamino-5-D-arginine vasopressin (DDAVP) on plasma cortisol (hydrocortisone). The influence of 1-deamino-8-D-arginine vasopressin (DDAVP), the new antidiutetic polypeptide without any side effects on plasma cortisol, was investigated in 30 healthy persons. A dose of 4 mug DDAVP administered intravenously induced a rise in plasma cortisol (hydrocortisone) levels greater than 3.5 mug/100 ml in 12 out of 20 persons studied. In this group (group I), the average increase at 15 minutes was 6.92+/-1.74 mug/100 ml (P less than 0.005), while in the remaining eight persons (group II) plasma cortisol levels decreased according to the usual normal daily rhythm. DDAVP, 80 mug, administered intranasally had no demonstrable influence on physiologic plasma cortisol regulation. On the basis of the present findings with relatively low doses, pituitary responsiveness (ACTH release) might be expected to occur in a higher percentage of persons after giving high intravenous doses of DDAVP. Further efforts are necessary to develop a safe vasopressin test for clinical examination of adenohypophyseal function.


Interactions of amygdala lesions with effects of pilocarpine and d-amphetamine on mouse killing, feeding, and drinking in rats. Repeated injections of 7.5 mg/kg pilocarpine induced mouse killing in both amygdala-lesioned and sham-operated rats, but more injections were required in the lesioned animals. Killing was evoked least readily in rats that showed substantial weight loss after surgery and that had damage to more medial regions of the amygdala. d-Amphetamine (.75, 1.50, or 3.00 mg/kg), administered either before or after a killing test, inhibited pilocarpine-induced killing in both surgical groups. Amygdala lesions attenuated pilocarpine-facilitated drinking in sated animals but did not alter the inhibitory effects of either pilocarpine or d-amphetamine on feeding or drinking.


A copulatory system in the hypothalamus and tegmentum of the domestic chick. A tract in the chick is described that runs from the posterior nucleus paraventricularis magnocellularis (PVM) through the lateral hypothalamus to a small area of the anterior tegmentum, which is adjacent to the ventral tegmental area. Bilateral lesions of the posterior PVM or of the PVM tract or of its tegmental destination abolish the facilitation of copulation in male chicks by testosterone. The effect is specific in that the facilitation of attack is either unaffected or only slightly depressed by such lesions, while being itself specifically depressed by other hypothalamic lesions. No other hypothalamic tracts were found that were necessary for the facilitation of copulation. An equivalent copulatory system probably exists in the mammal, with a more diffuse distribution in the preoptic area but with similar connections back to the tegmentum.


Another hamster paradox: more males carry pups and fewer kill and cannibalize young than do females. The response of virgin male and female golden hamsters to young was studied. In contrast to most species, males are more likely to carry pups than are females. All males carried pups, but approximately 50% of females cannibalized the young. The females that did not cannibalize the pups carried them with less hesitation and after shorter latencies than did the males. The response of females to young was not correlated with the aggressiveness displayed toward adult males during separate tests. Tests with gonadectomized females indicated that the maintenance of pup-killing behavior is not dependent on concurrent gonadal hormones. Progesterone injections did not significantly increase pup killing in males that had previously carried young. Speculations on the adaptive significance of the male and female hamster's response to pups are presented.


Cellular studies of mammary tissue from cows hormonally induced into lactation: lactose and fatty acid synthesis. Temporal changes in ability of mammary gland to synthesize lactose and fatty acids were identified during the treatment of cows hormonally induced to lactate, and animal differences were compared to subsequent milk production. Hormonal treatment involved 17 beta-estradiol + progesterone on days 1 to 7 and dexamethasone on days 17 to 19. Mammary tissue obtained by biopsy on days 0, 8, 16, and 26 of treatment was examined for biosynthetic capacity by tissue slice incubations. In terms of peak daily milk yield, one cow was very successful (greater than 30 kg), two were intermediate (9 to 10 kg), and one cow was unsuccessful (less than 3 kg). Differences between cows in the capability to synthesize lactose and fatty acids were evident as early as day 8 and were further magnified by day 16. In particular, the tissue from the successful cow was undergoing lactogenesis by day 8 while this was not evident until the day 16 biopsy sample in the less successful cows. In contrast to the other cows, tissues from the unsuccessful animal regressed in its ability to synthesize lactose and fatty acids between day 16 and 26. Relative differences between animals in measurements of metabolic capacity were consistent with subsequent milk production.


Cellular studies of mammary tissue from cows hormonally induced into lactation: histology and ultrastructure. Cytological changes were investigated in mammary parenchyma of cows hormonally induced to lactate. Four cows were induced with injections of 17 beta-estradiol plus progesterone (days 1 to 7) and dexamethasone (days 17 to 19). Tissue biopsies were obtained on days 0, 8, 16, and 26; and milking commenced on day 18 or 21 of the treatment period. Mammary tissue of cow A, the most successful animal, showed marked cellular proliferation and differentiation by day 8. By day 16, the parenchyma of cow A had undergone lactogenesis and exhibited histological and ultrastructural characteristics associated with actively secreting mammary tissue. Mammary epithelia of cow D, the least successful animal, showed minimal development by day 8 and ultrastructurally resembled nondifferentiated mammary cells. Mammary parenchyma of cow D had not undergone differentiation and was involuting by day 16. Mammary tissues of cow B and C showed intermediate responses with only slight development by day 16 and only limited areas of secretory activity by day 26. Results suggest that animals successfully induced into lactation undergo critical periods of cellular proliferation and differentiation which unsuccessful animals do not experience. Cytological differences were first evident by day 8, further magnified by day 16 of the treatment period, and subsequently expressed in terms of milk production (peak milk yield equaled 30.5, 10.0, 9.2, and 2.4 kg/day for cows A, B, C, and D).


Plasma progestins, estradiol, and luteinizing hormone following prostaglandin F2 alpha injection. Dairy animals, ranging from days 8 to 13 of the estrous cycle, were fitted with indwelling jugular catheters 1 day prior to either intramuscular injection of prostaglandin F2alpha free acid (30 mg, n=4) or intrauterine deposition of prostaglandin F2alpha free acid (10 mg, n = 3). Blood samples were collected at 6, 4, 2, and 0 h prior to administration of prostaglandin F2alpha and at 1, 3, and every 2 h thereafter until ovulation. Progestins, estradiol, and luteinizing hormone in plasma were measured by radioimmunoassay. Hormonal changes and interrelationships within animals were evaluated by least squares analyses. Decreases in progestins of plasma within 24 h indicated prostaglandin F2alpha induced luteolysis in six of the seven animals. Estradiol increased linearly from time of injection to 52 h postinjection. Intervals from administration of prostaglandin to onset of estrus, peak of luteinizing hormone, and ovulation were 74.9 +/- 21, 78.8 +/- 21, and 99.5 +/- 19 h.


Variability of Alternaria alternata: biochemical and immunological characteristics of culture filtrates from seven isolates. Biochemical variability of culture filtrates from the common allergenic mold Alternaria alternata was studied. Differences between culture filtrates from 7 different isolates and between 4 batches of culture filtrate from the same isolate were observed, suggesting the unreliability of presently employed biochemical methods in the routine standardization of mold allergens and the possibility of difficulties in developing standard techniques for their purification. Dextranases, cellulases, and agarases, found in one culture filtrate, may be a further source of problems in laboratory techniques employing A. alternata. The proportions of nitrogen:carbohydrate:dialyzed dry weight were different in culture filtrates from each isolate. Polysaccharides in culture filtrates from 3 of the 7 isolated contained glucose, mannose, xylose, galactose, and a fifth, unidentified sugar. Polysaccharides from one isolate lacked xylose, and the fifth sugar was not demonstrable in 4 of the isolates. Despite the biochemical differences, extensive antigenic cross-reactivity between different isolates was found in precipitin studies and inhibition of antibody binding.


Biological strandard for electron microprobe analysis of intracellular sodium concentration. A method is described for the determination of the intracellular concentration of sodium in individual cells using the electron microprobe analyzer. This method gives an accuracy equal to that obtained by using flame photometry on tissues with large cell populations. Intracellular sodium was precipitated in the cell by a fixative containing pyroantimonate. Cartilaginous needles from shark fins which were equilibrated in saline solutions of differing concentrations were used as biological standards.


Regulation of the immune system by synthetic polynucleotides. VI. Amplification of the immune response in young and aging mice. Model systems for study of the action of adjuvants in immunodeficient states were developed in 10- to 14-day-old BALB/aj mice and aging BALB/aj mice (12 to 16 months). With sheep red blood cells as antigen polyadenylicpolyuridylic acid complexes (poly A:U) were found to be stimulatory in both the neonatal and aging mice. The effect of poly A:U was similar to that seen when 5 X 10(6) thymocytes from immunologically mature mice were given with antigen. Cell-free supernatant fluids induced by incubation of poly A:U with thymocytes likewise were capable of restoring the number of antibody-forming cells to normalcy in aging mice.


Receptor-blocking factor present in immune serum resembling auto-anti-idiotype antibody. Previous studies have shown that rabbit antibody-forming cells in the primary and secondary response possess cell-associated antigen-binding receptors. In the present study, we demonstrate that a factor appears in the serum of rabbits following immunization which inhibits the antigen binding of up to 60% of the receptor-bearing antibody-forming cells in both the primary and secondary response. These observations were made on lymph node cells from rabbits primed with either sheep red blood cells (SRBC)3 or 3-nitro-4-hydroxy-5-iodophenylacetic acid coupled to keyhole limpet hemocyanin (NIP-KLH). The inhibitory activity is not associated with anti-SRBC or anti-NIP antibody. In the primary response to SRBC, the antigen binding by day 6 antibody-forming cells is inhibited by the autologous days 7 to 10 inactivated and absorbed serum. In the secondary response to SRBC, the inhibitory factor peaks in the serum around day 10. Later, in both the primary and secondary immune response to SRBC, the inhibitory activity of the serum decreases rapidly. In the primary response to NIP-KLH, the inhibitory activity of the immune sera increased from day 7 through day 14. The receptor-inhibiting factor is antigen specific since the serum from SRBC-primed rabbits inhibits SRBC binding by anti-SRBC antibody-forming cells, but it does not inhibit NIP binding by anti-NIP antibody-forming cells. Similarly, serum from NIP-KLH-primed rabbits inhibits NIP binding by anti-NIP antibody-forming cells, but does not inhibit the SRBC receptor on the anti-SRBC antibody-forming cells. The inhibition is not due to the presence of antihapten or anti-SRBC antibody competing with receptor sites, since the immune sera from one SRBC-primed animal inhibit antigen binding of its own antibody-forming cells, but do not inhibit the antigen binding of antibody-forming cells from other SRBC-primed rabbits.


Enzymatic degradation of tumor cells damaged by antibody plus complement. Rapid degradation of ascites tumor cells damaged by the action of antibody plus complement was found to be accomplished by all proteolytic enzymes active at physiologic pH that were tested. For three types of murine ascites tumor cells (Ehrlich ascites, sarcoma-180, and L1210 leukemia), this rate of degradation at low trypsin concentrations was proportional to a high power of enzyme concentration. This suggests that the simultaneous action of two or more enzyme molecules at adjacent cell surface sites is necessary. Cell degradation was assayed by determination of cell volume distribution with a Coulter multi-channel particle size analyzer. The present study may offer clues to in vivo mechanisms of cell degradation.


Quantitative analysis of brain galactosylceramides by high performance liquid chromatography of their perbenzoyl derivatives. A high performance liquid chromatographic (HPLC) method for analysis of galactosylceramides as their benzyol derivates has been devised. Samples containing 10-150 nmoles of monohexoslceramides are benzoylated by heating for 60 min at 60 degree C in 0.5 ml of 10% (v/v) Benzoyl chloride in pyridine. The products are purified by solvent distribution and analyzed by SPLC. The benzoylated cerebrosides with nonhydroxy fatty acids are separated from those with hydroxy fatty acids on a Zipax column with 7% ethyl acetate in hexane as a solvent and UV absorption at 280 nm is recorded. This isocratic procedure can be applied directly to chloroform-methanol procedure can be applied directly to chloroform-methanol extracts of adult brain with a relative standard deviation of 3.0% for cerebrosides with nonhydroxy fatty acids and 4.0% for cerebrosides with hydroxy fatty acids. Sulfatides do not interfere in the assay and can be converted to cerebrosides after desulfation by mild acid methanolysis. Benzoylated glucosyl- and galactosylceramides can be separated on a MicroPak NH2 column with 1.5% 2-propanol in cyclopentane as the chromatographic solvent.


Recovery of pituitary secretion of gonadotrophins and prolactin during re-feeding after chronic restricted feeding in female rats. The effects on reproductive function of restriction to one half Of the normal food intake for 30 or 60 days and subsequent unrestricted feeding were investigated in adult female Sprague-Dawley rats. Restricted feeding resulted in the cessation of oestrous cycles within 9 to 28 days, associated with decreased pituitary, ovarian and uterine weights. Pituitary content and concentration of FSH were increased by restricted feeding and the levels of FSH in 60 day underfed rats were about twice as high as those of the controls at dioestrus. There was little difference in pituitary LH content between the underfed groups and the controls at the end of restricted feeding. Pituitary LH concentration was significantly higher in rats underfed for 60 days than in the control rats. Pituitary prolactin content was one half and one third of that in control rats in rats underfed for 30 and 60 days respectively. Pituitary prolactin concentration was also decreased by restricted feeding. At the end of restricted feeding, no differences in serum LH and prolactin levels were found between the groups, whereas the serum FSH level in 60 day underfed rats was higher than that in the controls at dioestrus. After re-feeding, normal oestrous cycles returned within 3-5 days in almost all rats, regardless of the length of the previous cessation of oestrous cycles. Ptuitary contents of FSH and LH in underfed rats decreased after re-feeding following the return of oestrous cycles. The rate of decrease was much greater in 60 day underfed rats than that in 30 day underfed rats. On the other hand, serum levels of FSH, LH and prolactin in these underfed rats were increased by re-feeding and the levels in the evening of the first pro-oestrus were higher than those in the morning of dioestrus and pro-oestrus. Serum levels of these hormones increased more in 60 day underfed rats than in the other groups at any stage. After re-feeding, pituitary, ovarian and uterine weights increased and the uterine epithelial layer was clearly repaired in both underfed groups, although not always to the control levels by the first oestrus. The end-bud system of the mammary gland which degenerated during restricted feeding was comparable to that of the controls at the first oestrus after re-feeding in 30 day underfed rats, but not in 60 day underfed rats.


Immunological studies on the incorporation of radioactive leucine into sea urchin embryonic proteins: extractability and turn-over of newly synthesized proteins. This study was undertaken in order to investigate the turn-over of newly synthesized sea urchin embryonic proteins, the distribution of these proteins in the cell and their changes in solubility with time. Refined immunological methods providing unique possibilities for studies of individual proteins were utilized. The proteins were obtained in two soluble fractions: a concentrate of proteins from the cytosol was obtained by homogenization in a hypotonic medium and another fraction constituting membrane-associated proteins was solubilized by detergent treatment of the residual pellet; 75% of the total protein content in early and 60% in later developmental stages was thus solubilized. The proteins solubilized by these mild methods retained their antigenicity and could be analyzed by immunodiffusion methods. The fate of newly synthesized embryonic proteins was studied by incorporation of a radioactive amino acid using two different incubation procedures. The results of these studies indicated that newly synthesized proteins changed their solubility properties with time. Proteins were thus transfered from the soluble to the insoluble fraction. The findings that more proteins became insoluble during development mentioned above also supports such an interpretation. The synthesis of individual proteins was studied by immunodiffusion and autoradiography. Most of the antigens in the two soluble fractions were identical. Experiments with absorbed antisera revealed that only two to four antigens were present exclusively in one of the two soluble fractions. Three groups of antigens differing in regard to their metabolic activities were distinguished. Some antigens were labeled after both procedures without substantially increasing in concentration during development. Thus, their labeling was probably sustained by turn-over. Other antigens were labeled only after one of the two incubation procedures indicating large variations in regard to synthesizing rates of individual proteins.


Abnormalities in somite segmentation following heat shock to Xenopus embryos. The typical abnormality induced by a 15 min shock at 37degreesC is a single discrete length along the somite file within which segmental boundaries are absent or irregular. The two sides of the same embryo present a similar but not necessarily identical appearance. Usually all the embryos in a treated batch show abnormalities of similar severity. Survival of treated embryos, the details of the visible malformations, and temporal aspects of the phenomenon have been studied. The results indicate a temperature sensitive period that traverses the neurula, from head to tail at about the same rate as the somites form, but some hours beforehand. The temperature sensitive process is not associated with cell determination and differentiation, and there are reasons for thinking that the specification of the normal somite number occurs independently. The results are discussed in relation to Cooke & Zeeman's model of a wave front interacting with an oscillator.


Evolution of the nucleolar apparatus during oogenesis in Acipenseridae. Evolution of the nucleoli has been followed during oogenesis in the Acipenserid fishes, Acipenser ruthenus (the sterlet) and A. guidenstadti (the sturgeon) using light and electron microscopes. In the ovaries of adults, the oogonial nuclei usually have a single nucleolus with an adjacent mass of paranucleolar fibrillar material. The cytoplasm of the oogonia contains two dense bodies peculiar only to gonocytes, one being electron dense and containing RNA and the otherbeing electron-lucent and lacking RNA...


Cultural factors influencing the utilization or production of acetate by Butyrivibrio fibrisolvens. Utilization of acetate by four strains of Butyrivibrio fibrisolvens was influenced by the composition of their growth medium. Growth-limiting glucose concentrations, low availability of CO2 and the presence of sodium lactate all stimulated acetate uptake by three strains. The type strain, D1, utilized acetate if the concentration of acetate added to the medium was at least 15 mumol ml-1. In batch culture, all strains produced acetate before entering a phase of acetate uptake. Continuous-culture studies with one strain showed that acetate uptake was dependent upon growth rate. The amount of n-butyrate produced in batch or continuous culture was closely linked to the amount of acetate taken up.


Ultrastructure of mumps virus replication in organotypic cultures of hamster choroid plexus. Organotypic cultures of newborn hamster choroid plexus were inoculated with equal titre doses of newly isolated or hamster adapted strains of mumps virus. The ultrastructure of virus replication in choroid epithelial cells of the cultures was compared. No qualitative differences were observed; however, the adapted strain produced significantly greater numbers of virions and earlier destruction of the cultures. These findings are consistent with previous in vivo observations of the ultrastructure of the replication of these strains in the newborn hamster central nervous system. This in vitro study leads further support to the hypothesis that differences in the in vivo biological effects of the virus strains are primarily the result of virus-cell rather than virus-host interactions.


Multicore disease in twins.

Multicore disease in twins. Multicore disease in identical twin boys presented in infancy as generalized weakness and torticollis. Motor milestones such as sitting, standing, and running were delayed, although by the age of 6 years marked improvement in muscle strength had occurred. Serum enzymes were normal. Muscle biopsy revealed multifocal areas of decreased oxidative enzyme activity. Ultrastructurally, these areas were characterized by myofilament disruption and Z-band streaming.


Electrophysiological technique for evaluating lesions of the conus medullaris and cauda equina. The authors describe the use of evoked electromyographic responses recorded in the anal sphincter induced by stimulation of the bladder wall and urethra in evaluating lesions of the conus medullaris and cauda equina in 110 patients. This reflex response took effect by way of the pelvic nerves and cauda equina to the sacral cord where the pudendal nerve nucleus was activated, resulting in a contraction of the external anal sphincter. Various lesions along this pathway have been shown to produce either increased latencies and depressed responses or complete loss of response depending on the extent of the lesion. The correlation of results of this technique with clinical, myelographic, and operative findings indicate it to be a useful clinical tool.


A study of the nycthemeral cycle of behavioural temperature regulation in man. 1. Four human subjects were rendered hyperthermic and hypothermic by immersion in warm and cool water, at 02.00, 08.00, 14.00 and 20.00 hr. Bath and oesophageal temperatures and pulse rate were recorded. Temperature preference was determined by operant behaviour and vote. The core temperature set-point for behavioural thermoregulation was estimated from the behavioural results. 2. The results are in accord with those of previous studies of the nyethemeral cycling of autonomic responsiveness to heat and cold with a heating up phase before noon and a cooling down phase during the early night. 3. Subjective sensations and behavioural responses were also found to follow a nycthemeral cycle with a minimum before noon and a maximum at 20.00 hr. 4. The core temperature set point was 0-7 degrees C higher after noon than before noon with a small phase advance from resting core temperature. This result suggests that the nycthemeral cyclic change in body temperature is due to a nycthemeral cyclic change in the set-point near to which body temperature is kept by both autonomic and behavioural thermoregulatory responses.


[Relationship between potential and impedance change associated with excitation of the nerve (author's transl)]. The action potential or the potential change and the impedance change associated with excitation of the sciatic nerve and electric nerve model (ENM, the equivalent circuit of the nerve fiber), were recorded by means of AC bridge supplied of AC of 13 kHz. Records were taken in excitation at the stimulated site by the square current of different duration, exponentially increasing current of different increasing gradient. Records were also taken in propagated excitation, in excitation elicited in refractory period and at the polarized site of the inactive nerve and ENM. By observation of these records, the following results were obtained.1. With respect to potential changes in nerve, two kinds of changes were found, namely, the one was in cause and effect relationship with the impedance change and the other was without relation to impedance change. 2. In the course of the action potential, both kinds of potential changes were found.


Oxytocin analogues with combined high smooth muscle and negligible antidiuretic activities. Investigation of position 7 in neurohypophyseal hormones. The proline residue in position 7 of oxytocin occupies one of the four corner positions in the two beta turns proposed for the preferred conformation of the pituitary hormone. It has been suggested that synthetic modifications of the residues in these corner positions will yield analogues in which one or more of the biological activities of the parent hormone is highly accentuated in terms of potency relative to other activities. In a continued effort to test this hypothesis the following analogues of oxytocin were prepared: [7-glycine]oxytocin, [1-beta-mercaptopropionic acid,7-glycine]oxytocin, [7-alanine]oxytocin, and [1-beta-mercaptopropionic acid,7-alanine]oxytocin. These peptides were found to possess the following specific activities, respectively: rat uterotonic, 65 +/- 2, 355 +/- 3, 22 +/- 1, 123 +/- 4; avian vasodepressor, 5.3 +/- 0.8, 17 +/- 0.4, 4.8 +/- 0.1, 9.8 +/- 0.5; rat antidiuretic, less than0.01, 0.062, 0.081 +/- 0.01, 0.17 +/- 0.01; rat pressor, 0.3, 0.5, 0.4, 0.5 unit/mg. Thus the analogues retain high uterotonic activity but exhibit strongly diminished renal and vascular activities relative to oxytocin. Especially noteworthy is [1-beta-mercaptopropionic acid,7-glycine]oxytocin with its high uterotonic activity but very low antidiuretic and pressor activities. The activity profile of this analogue combined with the fact that it is only slowly enzymatically degraded warrants further investigations of this peptide for clinical applications.


Microviscosity of togavirus membranes studied by fluorescence depolarization: influence of envelope proteins and the host cell. The microviscosities of the hydrophobic regions of the membranes of intact Semliki forest and Sindbis viruses grown on BHK-21 cells, of liposomes derived from the extracted viral lipids, and of protease-treated virions were measured by fluorescence depolorization using the fluorescence probe 1, 6-diphenyl-1,3,5-hexatriene. The intact virus membranes were found to have a higher microviscosity than did virus-derived liposomes, indicating the viral envelope proteins contribute to microviscosity. However, protease-treated virus, devoid of protruding spikes but with residual lipophilic peptide tails, was found to have a microviscosity more similar to that of the intact virus than to that of protein-free liposomes. Sindbis virus grown in BHK-21 cells at 37 C had a much higher microviscosity than did Sindbis virus grown on Aedes albopicuts cells at 22 C. Sindbis virus grwon in A. albopictus and BHK-21 cells also gave higher microviscosity values than did the intact host cells. These data indicate that both the virion proteins and the cellular lipids selected during viral growth and maturation contribute to the increased microviscosity of togavirus membranes.


Urethrosphincteric reconstruction for congenitally absent urethra. Three female patients with complete urinary incontinence owing to congenital absence of the urethra associated with complex congenital anomalies involving the caudal end of the urogenital sinus are reported. A sphincteric tube constructed from a flap of the anterior bladder wall was positioned in place of the missing urethra. Abundance of circularly oriented fibers in this neourethra provided sphincteric function sufficient to maintain continence and eliminate the need for urinary diversion. Details of the congenital anomalies and reconstructive techniques are discussed.


Attenuation of pathogenicity of fowl plague virus by recombination with other influenza A viruses nonpathogenic for fowl: nonexculsive dependence of pathogenicity on hemagglutinin and neuraminidase of the virus. A number of antigenic hybrids of influenza A viruses were produced possessing either the hemagglutinin or the neuraminidase of fowl plague virus and the corresponding antigen derived from another influenza A virus. Other recombinants were obtained carrying both surface antigens of fowl plague virus but differing from the parent in certain biological properties. None of the recombinants isolated were pathogenic for adult chickens. Most recombinants obtained after crosses between reciprocal recombinants carrying both fowl plague virus surface antigens were also apathogenic for chickens. Using the same parent recombinants for double infection some of the progeny "back-recombinants" were pathogenic, whereas others were not. From these results it is concluded that the surface components do not by themselves determine the pathogenicity of influenza A viruses.


Late recurrence of testicular tumor (choriocarcinoma). A case is presented in which a large choriocarcinoma was excised from the contralateral retroperitoneal area 15 years after a radical orchiectomy for seminoma with elements of embryonal carcinoma.


Continuing medical education and the Professional Standards Review Organizations. The history of continuing medical education (CME) activities in the United States originates in the early years of this century. CME programs have increased markedly in recent decades, but these activities have come to be viewed from many perspectives as inadequate. Recently, the Professional Standards Review Organization (PSRO's) have been proposed as a "solution" to the problems of CME. PSRO's may provide useful data to CME programs and could serve as mechanisms for evaluating these activities. Features of medical care problems which identify them as suitable subjects for CME activities using PSRO data are presented and discussed.


Lipid synthesis by perfused lung.

Lipid synthesis by perfused lung. An isolated lung ventilated with pulses of negative pressure and perfused through the pulmonary vasculature was utilized for the study of 3-sn-phosphatidylcholine synthesis. The perfusion fluid consisted of a Krebs-Ringer phosphate buffer with 6% bovine serum albumin, pH 7.4, and the appropriate substrate. The simultaneous incorporation of (1-14C) palmitate and (2-3H) glycerol and the simultaneous incorporation of (CH3-14C) choline and (CH3-3H) methionine were examined. From these experiments it is concluded: 1) lung tissue incorporates (2-3H) glycerol into 3-sn-phosphatidylcholine to a greater extent than any other lipid examined; 2) both choline and methionine contribute to the synthesis of 3-sn-phosphatidylcholine, and 50-70% of the label in its nitrogen base is derived from choline and 30-50% from methionine; and 3) a high PO2 appears to reduce the synthesis of 3-sn-phosphatidylcholine.