The binding of lectins to components of plasma membranes from porcine submaxillary lymph node lymphocytes. By sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis the plasma membranes from porcine lymphocytes contain at least 30--35 glycopolypeptides and one or more glycolipids to which one or more of 12 purified lectins bind. The specificities of binding generally followed the same pattern as those of the reaction of the lectin with intact pig lymphocytes. Some lectins (e.g., the isolectin pair, Agaricus bisporus lectins A and B and a group consisting of the Lens culinaris A and B isolectins and the closely related Pisum sativum lectins) bind to almost identical populations of plasma membrane components and compete with each other for all their binding sites. Others (e.g., Concanavalin A and the Lens culinaris-Pisum sativum group and a group consisting of phytohemagglutinin-L, Ricinus communis lectin-60 and Ricinus communis lectin-120 bind in a cross reactive manner to some common binding moieties but, in addition, to certain nonshared ones. Still others (e.g., soybean agglutinin, peanut agglutinin and wheat germ agglutinin) do not share any common binding moieties with the other lectins. The amount of lectin binding and the number of membrane components to which a lectin binds is directly related to the Ka of binding of the lectin to the intact lymphocyte. Those with high Ka (Cocanavalin A Lens culinaris lectins, Pisum sativum lectins, phytohemagglutinin-L), bind to 20-30 different components giving very complex binding patterns while those with lower Ka (Agaricus bisporus lectins, wheat germ agglutinin, peanut agglutinin, and soybean agglutinin) bind to 8--13 components with easily distinguishable patterns. Soybean agglutinin binds almost exclusively to a glycolipid fraction while for the others one or more glycopolypeptides served as the major lectin-binding molecule. The Ricinus lectins, two lymphocyte toxins, bind to essentially every plasma membrane component to which the mitogen phytohemagglutinin-L binds, in fact competing for most of those plasma membrane moieties which bind phytohemagglutinin-L.


Separation of transcriptively active and inactive chromatin. Agarose gel chromatography. Sheared chromatin fractionated by currently accepted methods of agarose gel exclusion chromatography, undergoes a limited and non-specific aggregation resulting from the high ionic strength and divalent cation concentration of the column elution buffer. Such aggregation causes the artifactual appearance of radioactively labeled, newly synthesized RNA within the column exclusion volume, erroneously suggesting an enrichment for actively transcribed chromatin. Claims for the efficacy of agarose gel exclusion as a method for separating template-active and -inactive chromatin are based largely on assays for active chromatin which rely on localization of specific molecular complexes of chromatin and nascent RNA. Under the conditions employed, the present studies invalidate this assay and thus cast considerable doubt on the agarose gel exclusion method itself.


Sex-related differences in diurnal activities and development of hepatic microsomal 3-hydroxy-3-methylglutaryl coenzyme A reductase and cholesterol 7alpha-hydroxylase. The activities of hepatic microsomal 3-hydroxy-3-methylglutaryl CoA reductase and cholesterol 7alpha-hydroxylase were consistently higher (up to 3-fold) in female compared to male rats fed 2% cholestyramine for 8 h daily. In all animals studied, enzymic activities were highest 6 h after feeding began. However, 85% of the rise in cholesterol 7alpha-hydroxylase activity occurred in the 6 h before and 89% of the rise in 3-hydroxy-3-methylglutaryl CoA reductase activity occurred in the 6 h after feeding started. Sex-related differences in both enzymic activities first became apparent at the time of sexual maturity. Enzymic activities before weaning were generally low and a late-suckling (13--20 days) rise in cholesterol 7alpha-hydroxylase was not accompanied by a rise in 3-hydroxy-3-methylglutaryl CoA reductase. For all of these studies we assayed cholesterol 7alpha-hydroxylase at two concentrations of exogenous cholesterol to obviate problems relating to size of the cholesterol pool.


Changes in the phospholipid molecular species composition of soybean hypocotyl and cotyledon after dedifferentiation. The effect of dedifferentiation on the molecular species composition of soybean phospholipids was studied by using hypocotyl, cotyledon and the suspension culture cells established from those organs. Three major phospholipids (phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol) and phosphatidylmonomethylethanolamine were composed of twelve molecular species. Major species were 1-palmitoyl-2-linoleoyl, 1-obeoyl-2-linoleoyl, 1-palmitoyl-2-linolenoyl and 1-linoleoyl-2-linoleoyl species. Different proportions of the molecular species were found among the three major phospholipids, but phosphatidylmonomethylethanolamine was composed of the same proportions of the molecular species as those of phosphatidylethanolamine. After dedifferentiation, the 1-palmitoyl-2-linoleoyl species increased in the cell established from hypocotyl. In the cells established from cotyledon, the 1-palmitoyl-2-linolenoyl species increased dramatically. In both cells, the 1-palmitoyl-2-linolenoyl species increased in response to increase in the 2,4-dichlorophenoxyacetic acid concentrations and the progress of cell growth.


Synthesis of lipoprotein lipase in cultured avian granulosa cells. Avian granulosa cells cultured as a homogeneous parenchymal population contain lipolytic activity. This activity is stimulated 2--5-fold by serum, inhibited 90% by 1 M NaCl and inhibited 80% by specific anti-lipoprotein lipase immunoglobulins. 85% of the activity binds to heparin-Sepharose 4B, and 70% of bound activity is eluted with 1.5 M NaCl. Thus, the lipolytic activity of cultured granulosa cells is lipoprotein lipase. Granulosa cells were shown to synthesize lipoprotein lipase in culture by incorporating [3H]leucine into the enzyme protein, as measured with an immunoadsorption technique. Finally, colchicine was shown to increase intracellular lipolytic activity, suggesting an inhibition of secretion of this enzyme by cultured granulosa cells.


The monomeric casein composition of different size bovine casein micelles. The fractionation by size of casein micelles from bovine skim milk was performed by chromatography on controlled-pore glass granules (CPG-10/3000). Acid precipitation of the fractionated proteins in combination with polyacrylamide gel electrophoresis gave no indication for monomeric caseins in the whey fractions. A factor besides low temperature appears necessary for the dissociation of, for example, beta-casein from casein micelles. The casein composition was studied by DEAE-cellulose chromatography. In bulk skim milk the alphas-, beta- and kappa-caseins were shown to occur in the following relative amounts: 52, 33 and 15%, respectively. The distribution varies with the size of the micelle. In large and medium size micelles the alphas1-casein content is almost constant; beta-casein and kappa-casein appear to be complementary so that the kappa-casein content increases with the decrease in the size of the micelle. In small micelles the relative beta-casein content is about 50%, alphas1-casein is only about 33%. We suggest that beta-casein plays a special role as initiator of micelle formation, and that alphas1-casein stabilizes the structure of the larger micelles.


Serum albumin domain secondary structure prediction. A new method has been used to predict probability profiles for helix, beta-sheet and bend structures along the entire sequence and derive an averaged profile for the three homologous domains. The results are correlated with the disulphide bridge pattern, the distribution of hydrophobic sites and points where albumin is cleaved by enzymes.


Flexibility of bovine pancreatic trypsin inhibitor. The native conformation of a protein may be expressed in terms of the dihedral angles, phi's and psi's for the backbone, and kappa's for the side chains, for a given geometry (bond lengths and bond angles). We have developed a method to obtain the dihedral angles for a low-energy structure of a protein, starting with the X-ray structure; it is applied here to examine the degree of flexibility of bovine pancreatic trypsin inhibitor. Minimization of the total energy of the inhibitor (including nonbonded, electrostatic, torsional, hydrogen bonding, and disulfide loop energies) yields a conformation having a total energy of -221 kcal/mol and a root mean square deviation between all atoms of the computed and experimental structures of 0.63 A. The optimal conformation is not unique, however, there being at least two other conformations of low-energy (-222 and -220 kcal/mol), which resemble the experimental one (root mean square deviations of 0.66 and 0.64 A, respectively). These three conformations are located in different positions in phi, psi space, i.e., with a total deviation of 81 degrees, 100 degrees and 55 degrees from each other (with a root mean square deviation of several degrees per dihedral angle from each other). The nonbonded energies of the backbones, calculated along lines in phi, psi space connecting these three conformations, are all negative, without any intervening energy barriers (on an energy contour map in the phi, psi plane). Side chains were attached at several representative positions in this plane, and the total energy was minimized by varying the kappa's. The energies were of approximately the same magnitude as the previous ones, indicating that the conformation of low energy is flexible to some extent in a restricted region of phi, psi space. Interestingly, the difference delta phi i+1 in phi i+1 for the (i + 1)th residue from one conformation to another is approximately the same as -delta psi i for the ith residue; i.e., the plane of the peptide group between the ith and (i + 1)th residues re-orient without significant changes in the positions of the other atoms. The flexibility of the orientations of the planes of the peptide groups is probably coupled in a cooperative manner to the flexibility of the positions of the backbone and side-chain atoms.


Cardiac and forearm plethysmographic responses to high intensity auditory stimulation. This paper reports an investigation of forearm blood flow and cardiac responses to high intensity auditory stimulation. Blood was assessed in terms of forearm girth (FG) using a strain gauge, and since this technique had not been used previously, a preliminary study was conducted to validate the measure. In Experiment 1 (N = 24), subjects performed either a fast- or slow-paced mental arithmetic task. The data indicated that the strain gauge technique differentiated periods of rest from arithmetic stress and produced results comparable with those obtained using limb volume plethysmography. In Experiment 2 (N = 24), subjects received eight presentations of either a 60 dB or a 110 dB white noise stimulus at randomly ordered intervals of 35, 40, 45 and 50 sec; stimulus rise time was 50 msec and the duration 1 sec. Both groups displayed short-latency (i.e. within 10 beats poststimulus) cardiac accelerative responses which habituated over trials. In addition, the 110dB group displayed a long-latency (19.9 sec) accelerative response of approximately 25 beats per min and this was accompanied by an increase in FG. These responses occurred only following the first stimulus presentation, and analysis of the EKG T-wave amplitude suggested that the cardiac response was mediated sympathetically. These results are discussed in terms of conceptions of the startle and defence responses in man and the fight/flight reaction in animals.


Maintenance requirements for bacteria growing on C1-compounds. The maintenance coefficient, ms (mmol substrate/g cell dry wt hr), of two distinct groups of C1-utilizing bacteria has been determined by growing the organisms in an aerobic continuous culture limited by different C1 growth substrates. For growth on methanol, ms = 2.5 +/- 0.3 for Pseudomonas C; 3.9 +/- 0.7 for Ps. methylotropha (these bacteria utilize methanol via the ribulose monophosphate pathway of formaldehyde fixation); 1.5 +/- 0.2 for Pseudomonas 1, and 2.3 +/- 0.4 for Pseudomonas 135 (the latter bacteria utilize C1-compounds via the serine pathway). For growth on formaldehyde, ms = 1.5 +/- 0.3 for Pseudomonas 1 and 2.7 +/- 0.7 for Pseudomonas 135, whereas on formate the values for ms are 1.0 +/- 0.2 and 4.4 +/- 1.3; respectively. Although the maintenance coefficients did not differ systematically between the two groups of bacteria, the maintenance requirements per generation of the serine pathway bacteria were considerably higher (8.7 vs. 3.9) owing to their slower growth rate. The maximum molar yield values, YMmax (g cell dry wt/mol substrate utilized), corrected for the maintenance energy of bacteria which utilize C1-compounds via the ribulose monophosphate pathway averaged 19.1 when grown on methanol, while the values for bacteria which use the serine pathway averaged 13.5. On formaldehyde an average value of 11.5 is obtained and on formate the average value was 7.4 in the serine pathway bacteria.


[Transplacental effect of urethane on DNA synthesis in mouse embryonal lung tissue during organotypical cultivation]. Kinetics of DNA synthesis was estimated by autoradiography in the organ cultures of murine embryonic lung tissue. Strain A mice were taken for this experiment (intact and exposed to transplacental action of urethane--30 mg/g to a pregnant mouse, subcutaneously, once). The explants were examined 1,7, 15 and 21 days after the cultivation. Transplacental urethane inhibited DNA synthesis the first 24 hours. The label index became approximately 3 times less. The next two days it was restored and then stimulated. The maximal label index was noted in the experimental explants on the 7th cultivation day, i.e. on the 8th-10th day after the transplacental action of urethane on the lung tissue in utero. By the 15th cultivation day the label index dropped but was higher than in the intact explants; by the 21st day DNA synthesis in the intact cultures ceased completely, whereas in the experimental animals--it continued.


[Immune response in rats and mice with adjuvant disease]. There were phasic changes of immune response in rats and mice with adjuvant disease: an increase of antibody production on the 7th day after the injection of complete Freund's adjuvant and its depression on the 21st day; there was also observed suppression of normal antibody production to O- and Vi-antigens of typhoid bacilli.


Antidiuretic effect of morphine in the rat: tolerance and physical dependence. 1 Injection of rats with morphine or methadone, before they received a water load equivalent to 5% of their body weight, produced a dose-dependent antidiuretic effect. Following the antidiuresis, urine was eliminated with kinetics similar to control untreated rats. 2 The antidiuretic effect of morphine or methadone was blocked by naloxone administered before the opiate, or reversed when given after the opiate. 3 Rats implanted with morphine pellets developed a marked degree of tolerance to the antidiuretic effect of morphine. Tolerance was also obtained on injection of three daily doses of morphine or methadone over two days. 4 Withdrawal symptoms were precipitated by naloxone in rats implanted with pellets of morphine; under these conditions the animals showed a marked reduction in urine production as compared to naive rats.


The effect on the bladder pressure of sudden entry of fluid into the posterior urethra. The posterior urethra was stimulated by the rapid injection of saline against the urethral wall. The effect was studied in 50 female patients attending a urodynamic clinic. The injection was made via a double lumen catheter, and bladder, urethral and rectal pressures were observed during and after the ingress of fluid. The results indicate that sudden entry of fluid into the posterior urethra does not initiate a detrusor contraction even in those patients in whom the injection of fluid is painful. The relevance of these results in relation to the unstable bladder and urodynamic tests is discussed.


Variations in serum dopamine beta-hydroxylase in normal subjects and chronic alcoholics. Serum dopamine beta-hydroxylase (DBH) activity varies greatly between individuals but is usually relatively constant within individuals. DBH activity was determined in 20 normal subjects and 6 chronic alcoholics during alcohol ingestion and withdrawal, under controlled and standardized conditions. For all subjects mean random DBH was 423 +/- 249 (mean +/- SD) nmol phenylethanolamine/h per millilitre serum. Between-day serum DBH values vary more than within-day values (21.1% vs 15.1%). Cold-pressor testing or sudden standing does not increase mean DBH; however, some individuals show a significant increase which cannot be elicited on repeat testing. Mean DBH activity did not vary significantly over 24 h. Clinically useful correlations between single random DBH and blood pressure or 24-h urine catechols should not be expected.


Microbes associated with the epithelium of the oesophagus and stomach of guinea pigs (Cavia porcellus). Histological examination of oesophagus and stomach specimens from 28 guinea pigs (Cavia porcellus) revealed microbes associated with the epithelial surface of these organs. Layers of Gram-positive cocci were associated with oesophageal tissue in all the animals. Yeasts (Torulop-sid pintolopesii) were associated with stomach tissue in most animals 1 week of age or older.


Metastatic adenocarcinoma to the anterior uvea and elevated CEA levels. A 47-year-old man developed chronic unilateral iridocyclitis with secondary glaucoma, and elevated plasma CEA levels. The eye was enucleated seven months later, and histopathologic examination revealed a metastatic mucin producing CEA positive adenocarcinoma. The patient died one year after the initial onset of symptoms. The primary site of the tumor was not proven, but most likely was the gastrointestinal tract.


The immunochemistry of lambda-type carrageenans from certain red algae. An antibody preparation directed against a structural feature associated with 6-sulphate groups was used to probe structural relations among certain lambda-type carrageenans. Immunochemical and chemical differences are described between the KC1-soluble carrageenans from tetrasporic algal plants of Gigartina corymbifera, Gigartina sp. from San Francisco Bay, Petrocelis middendorfii, Iridaea cordata, Rhodoglossum californicum, and Chondrus crispus. The differences in immunochemical reactivity of the Gigartina and Petrocelis carrageenans relative to the homologous antigen (Chondrus crispus lambda-carrageenan) are attributed to the lower content of 6-sulphate groups on the 4-linked residues in the former carrageenans. Both the immunochemical and chemical data suggest that the Gigartina and Petrocelis carrageenans are largely xi-like in structure but do contain lambda-like features. The i.r. spectrum of the Petrocelis carrageenan differs from that of the Gigartina carrageenans. The carrageenans from I. cordata and R. californicum differ to a lesser degree from Chondrus crispus lambda-type carrageenan. These differences cannot be accounted for by differences in the levels of 6-sulphate groups. Some other structural feature, as yet unidentified, is responsible for the discrepancy in the immunochemical reactivity of these carrageenans to the anti-lambda-carrageenan.


A study of the specificity of Bandeiraea simplicifolia lectin I by competitive-binding assay with blood-group substances and with blood-group A and B active and other oligosaccharides. The specificity of Bandeiraea simplicifolia lectin I (BS I) has been studied by competitive-binding assays (CBA) using tritium-labeled human B and hog A substances. Blood-group B substances isolated from horse gastric mucosae and from human ovarian-cyst fluids were much better inhibitors of binding of tritiated blood-group B substance to insoluble BS I-Sepharose 2B than were human blood-group A substances from saliva and ovarian-cyst fluid. A and B active blood-group substances showed the same range of potency in inhibiting binding of tritium-labeled hog A substance to BS I-Sepharose 2B. CBA with BS I-Sepharose 2B, labeled human blood-group B substance, and human blood-group A and B active aligosaccharides separated the haptens into two groups differing in slope. Group 1, containing methyl alpha-D-GalNAcp, D-GalNAcp, and an A active pentasaccharide ARL 0.52, with 3, 19, and 25 nmol respectively needed for 50% inhibition of binding, has a lower slope than group 2, which contains alpha-D-GalNAcp-(1 leads to 3)-2-acetamido-2-deoxy-D-galactitol and p-nitrophenyl alpha-D-GalNAcp, with 3 nmol of each required for 50% inhibition of binding, as well as ten glycosides with terminal, nonreducing, alpha-linked D-Galp. The most potent inhibitors of this group were p-nitrophenyl alpha-D-Galp, alpha-D-Galp-(1 leads to 3)-D-Galp, alpha-D-Galp-(1 leads to 6)-D-Glcp, and methyl alpha-D-Galp, with 5, 7.4, 9.6, and 11 nmol respectively needed to inhibit binding by 50%. The difference in slopes was explainable in terms of a recent finding that BS I exists as a mixture of five isolectins composed of two subunits having different specificities; subunit A is most specific for alpha-linked, terminal, nonreducing D-GalNAcp, but it also reacts with alpha-linked, terminal, nonreducing D-Galp, whereas subunit B tends to be more specific for terminal, nonreducing, alpha-linked D-Galp.


Microtubules in mouse embryo fibroblasts extracted with Triton X-100. Treatment of mouse embryo fibroblasts with 1% Triton X-100 at 37 degrees C in the presence of 4M glycerol and 1 mM EGTA results in the extraction of about 80% cellular proteins. Indirect immunofluorescent staining with monospecific antibodies against tubulin showed that extracted cultures contained a well developed system of cytoplasmic microtubules, indistinguishable from a system of control non-extracted cells. Microtubules in extracted cells were sensitive to Ca2+ ions, and to cold or prolonged incubation in a glycerol-free buffer. Sodium dodecylsulphate-polyacrilamide gel electrophoresis revealed proteins co-electrophoresed with tubulin and actin in Triton-treated cultures. Electron microscopy demonstrated the presence of both microtubules and microfilament bundles in the extracted cells, but complete dissolution of plasma and intracellular membranes.


Iron transport in NRK cells synchronized in G(1) by picolinic acid. NRK cells treated with picolinic acid are arrested in G(1)(G0). After removal of the agent a wave of transferrin-dependent iron uptake was observed only in early G(1). Therefore, iron transport is a primary event in the reinitiation of cell growth and may be connected with subsequent metabolic steps leading to initiation of DNA synthesis.


Stimulation by glucocorticoids of myoblast growth at low cell densities. Proliferation of rat myoblasts was stimulated by addition of 10(-7) M dexamethasone to cells plated at 25 cells/cm(2). In the presence of fetal calf serum and chick embryo extract, physiological levels of glucocorticoids caused a 3- to 7-fold increase in colony formation. In mass cultures, proliferation of myoblasts was also stimulated by dexamethasone, but fibroblast growth was inhibited. These opposite effects of glucocorticoids on myoblast and fibroblast growth suggest their use in cloning muscle cells from mixed cell populations.


Supranucleosomal structure of chromatin. Rat liver chromatin was moderately digested by micrococcal nuclease and analysed by centrifugation in isokinetic sucrose gradients and electron microscopy. Two classes of particles sedimenting with about 33S and 60S were characterized. Kinetics of their appearance and disappearance during progressive digestion suggests that they represent monomers and dimers cleaved from a higher order (supranucleosomal) structure of chromatin. Biochemical and electron microscopical results suggest that the monomers and dimers contain eight and sixteen nucleosomes, respectively, which are densely packed into 23 nm (monomer) and 29 nm (dimer) globules.


Relationship between the results of bronchial, nasal and conjunctival provocation tests in patients with asthma. Skin tests and bronchial, nasal and conjunctival provocation tests with pollen and animal dander allergens were performed in thirty patients with atopic asthma. In vivo test results were compared only when the same batch of allergen had been used. A nasal reaction was mostly elicied at a lower concentration of allergen than was needed to elicit a bronchial reaction (P less than 0.01). Positive nasal reactions were often obtained when the corresponding bronchial tests were negative. The conjunctiva reacted to lower concentrations than the bronchi in a third of the instances of testing, with most of these being tests with mugwort allergen (P less than 0.05). A positive skin test in cases with a negative bronchial challenge test was often accompained by a positive nasal test and in some cases by a positive conjunctival test. It is concluded that nasal or conjunctival provocation tests do not replace bronchial challenge tests. In an asthmatic patient who gives no reaction to bronchial challenge with a particular allergen, a positive skin test may reflect a nasal allergy.


Treatment of infantile atopic dermatitis with a strict elimination diet. Of twenty-one infants with atopic dermatitis, twenty were treated with a strict elimination diet for a period of up to 6 weeks. Seven infants healed and twelve improved. The infants who healed were less than 6 months old and had had a short duration of dermatitis. For one infant the skin condition was unchanged. Another infant was breast-fed throughout the period with dermatitis. Blood eosinophilia and/or elevated serum IgE commonly found on admission decreased significantly (P less than 0.01) during the diet period. On challenge with cows' milk, twelve infants were considered as intolerant. At the age of 2 years the dermatitis had cleared in all but four children. Of these four children, two were still cows' milk intolerant. Another two infants were also cows' milk intolerant, but without dermatitis.


Aerobiological studies based in Derby. II. Simultaneous pollen and spore sampling at eight sites within a 60 km radius. In order to determine whether a sampling site in the centre of Derby, where air sampling had been carried out for five summers, really provided valid aerobiological data for the surrounding area, eight identical volumetric spore traps were operated simultaneously during the summer of 1969 at various sites up to 56 km from Derby. For most pollen and spore types, total numbers and seasonal pattern at all the sampling sites were found to be similar, although a few extreme and unexpected variations were noted. Figures varied somewhat in different types of site, and local effects are emphasized.


Evaluation of a direct radioimmunoassay for carcinoembryonic antigen. 1. A radioimmunoassay for CEA using commercially available reagents (CEA-Roche), which avoids the perchloric acid and dialysis steps used in the CEA-Roche assay, has been devised. 2. Comparison of results on 746 plasma samples using our assay with those obtained using the CEA-Roche assay revealed a highly significant correlation coefficient (0.72; p less than 0.00001). 3. A further comparison of results on 373 plasma samples using our assay, but including a preincubation step with goat serum, with those obtained using the CEA-Roche assay also revealed a highly significant correlation coefficient (0.89; 0.00001). The goat serum preincubation step eliminated most of the discordant results in which our assay value was higher than CEA-Roche assay, presumably because of anti-goat antibodies in the patient plasma sample.


Experimental optimization of the detection limit of one-step solid-phase radioimmunoassay. The minimal detection limit and the conditions of maximal sensitivity of a one-step solid-phase inhibition radioimmunoassay for human immunoglobulin A have been determined by application of statistical methods of experimental optimization. The choice of the optimal combination of qualitative variables, such as the origin of the antibody and the nature of the solid phase, was made by the study of a covariable under non-optimal conditions of the quantitative variables, such as the amount of antibody. The covariable was the avidity of the antibody, which is expected to have a large influence on the sensitivity. Only the difference in avidity between two immunosorbents with cellulose or Sepharose as solid-phase material proved to be statistically significant, and further study was done with cellulose. The experimental optimization of the sensitivity as a function of five quantitative variables yielded a reduction of the detection limit by a factor 5.6 (from 23.5 to 4.2 ng IgA). The variables determining the amount of insolubilized antibody in the assay had the largest influence on the value of the detection limit. The conditions of optimal sensitivity did agree with the predictions by a physical model of radioimmunoassay. The results are discussed in relation to the assay parameters such as the amount and the avidity of the insolubilized antibody and the initial percentage of binding, and in relation with theoretical optimization of the sensitivity.


A simple technique for the measurement of ouabain-sensitive sodium transport in red cells. A new, simple and non-radioactive technique is described for the measurement of ouabain-sensitive sodium transport in red cells, which is an alternative to the more complex standard technique using radioactive sodium. The new technique is based on the observation that when excess ouabain is added to blood, the sodium content of the red cells rises, due to complete inhibition of ouabain-sensitive sodium efflux while influx remains unchanged. The rate of increase of red cell sodium is therefore a measure of ouabain-sensitive sodium efflux (Eos). This estimate of Eos, when expressed as a fraction of the original red cell sodium concentration, gives a measure of the ouabain-sensitive, efflux rate constant (ERCos). In healthy persons, ERCos obtained with this new technique agrees closely with that obtained with the standard radioactive technique. In hypokalaemic patients the ERCos estimated by the new technique was less than that obtained with the standard technique, but this difference disappeared as the patients became normokalaemic. We suggest that this difference arises because the new technique measures only net ouabain-sensitive sodium transport whereas the standard technique also measures the ouabain-sensitive Na : Na exchange which is present in patients with hypokalaemia.


Preparation of 125iodine-labelled methotrexate and its use in a magnetisable particle solid-phase radioimmunoassay. Two techniques for the iodination of methotrexate are describes, involving covalent linkage to the drug of 125I-labelled N-succinimidyl-3-(4-hydroxyphenyl)propionate (Bolton and Hunter reagent), or to 125I-labelled histamine. A rapid highly specific radioimmunoassay for methotrexate was developed, employing a specific antiserum covalently linked to magnetisable particles, and 125I-labelled methotrexate as tracer. Incubation times for the assays were 60 and 10 min for the Bolton and Hunter reagent-linked methotrexate and 125I-labelled histamine-linked methotrexate respectively. Separation of bound from free antigen was achieved by a rapid magnetic separation system. Results obtained for serum samples correlated closely with those using an enzymatic (dihydrofolate reductase) competitive protein binding assay for methotrexate. A major advantage of the assay is its potential for processing large numbers of samples rapidly, making it highly suitable for routine clinical use.


Immunology of the lower respiratory tract. III. Concentrations of antigen and of antibody-forming cells in pulmonary and systemic lymphoid tissues of dogs after intrapulmonary or intravenous administration of sheep erythrocytes. The distribution of antibody-forming cells to sheep erythrocytes among canine pulmonary and systemic lymphoid tissues differs distinctively depending on whether antigen is administered by the intrapulmonary or the intravenous (i.v.) route. After local (intrapulmonary) immunization, antibody-forming cells are restricted to the lung and regional lymph nodes; after i.v., they are widespread. To test the hypothesis that the tissue distribution of antigen is an important determinant of the resultant distribution of antibody-forming cells, dogs were immunized with radioiodinated sheep erythrocytes, either intrapulmonary (i.p.) or i.v. After 4 days, the concentrations of tissue-bound radioactivity and of antibody-forming cells in various lymphoid tissues were compared. The distribution of tissue-bound radioactivity among lymphoid tissues was clearly determined and different depending on the route of immunization. After i.p. administration, radioactivity was bound to lung and hilar lymph nodes; after i.v., it was found in the lung, liver, spleen and occasionally hilar nodes. Antibody-forming cells appeared in those lymphoid preparations which contained tissue-bound radioactivity. The exception was that locally applied antigen elicited antibody-forming cells in the lung poorly, despite the fact that abundant antigen remained localized to the lung. Notably, i.v. immunization resulted in the appearance of both antibody forming cells and cell-bound radioactivity in the lung. It is concluded that the distribution of antigen among tissues is a major determinant of the distinctive patterns of appearance of antibody-forming cells after different routes of immunization.


Osteogenesis imperfecta in one of twins. Case report. The is a report of a 5-year-old girl with osteogenesis imperfecta, an uncommon abnormality considered to encompass several distinct disorders and biochemically classified as a connective tissue disease. Affected individuals, in the more severe form of the disease, often die in utero or shortly thereafter secondary to birth trauma. The patient was one of dizygotic twins discordant for osteogenesis imperfecta tarda inherited as an autosomal dominant trait.


Remote cerebral hemisphere symptoms from surgically treated patients with posterior fossa brain tumors; vascular factors: a basis for a theory concerning space sickness. Three case histories of patients with large tumors in the posterior fossa who were operated on in a sitting position subsequently developed 1 or more symptoms referable to the temporoparietooccipital regions of the brain 24 to 48 hours postoperatively. Initially, it was believed that such symptoms were due to a stimulation of the association pathways causing firing of remote association areas (See Ch. 4). Subsequent studies of the rotation of blood vessels of the brain in the developing embryo and a review of the anatomical location of the arteries supplying the temporoparietooccipital region led to the conclusion that some compromise of the posterior cerebral artery was responsible for the symptoms. The symptomatology in these brain tumor patients was not unlike that seen in the cosmonauts and astronauts in space flight, designated as "motion sickness" in the space literature. A suggestion was made as to clarification of the definitions. This author advocated that the term "motion sickness" be confined to those symptoms of dizziness, nausea, and vomiting, due to involvement of the peripheral end organ, the inner ear. "Space sickness" might include these symptoms but also might have the addition of disorientation or the inversion of image in space and formed or unformed hallucinations. These relate to the temporoparietooccipital area, the midtemporal, and the occipital regions. In such instances, there must be central involvement or a stimulation of this interpretive cortex of the brain. The remote symptoms from the supratentorial cotex were believed to be due to hypoxia related to the posterior cerebral artery compromise, resulting in delayed "luxury perfusion" and the development of local lactic acidosis. Transaxial transmission of force with an uncal tentorial herniation causing compression of the posterior cerebral artery was suggested as a mechanism responsible for the vascular compression.


Diurnal rhythms of aminopyrine metabolism: failure to sleep deprivation to affect them. After a single oral dose of aminopyrine (9 mg/kg), mean salivary aminopyrine half-lives (t 1/2s) and metabolic clearance rates in 12 normal male volunteers exhibited diurnal variations. Salivary aminopyrine t 1/2s were approximately 50% longer at 8 P.M. (2.1 +/- 0.7 hr) than at 8 A.M. (1.4 +/- 0.3 hr). Mean aminopyrine metabolic clearance rates decreased 20% from 8 A.M. (418.2 +/- 152.0 ml/min) to 8 P.M. (335.3 +/- 107.6 ml/min). There were large interindividual variations in the magnitude of these diurnal changes in aminopyrine t 1/2 and metabolic clearance rates. There were nonsignificant changes in mean aminopyrine apparent volumes of distribution (aVd) which increased only slightly from 53.1 +/- 20.6 L at 8 A.M. to 59.7 +/- 26.5 L at 8 P.M. There were diurnal variations in plasma 11-hydroxycorticosteroids (11-OHCS), mean values of which decreased 60% from 16.2 +/- 4.2 microgram/100 ml at 8 A.M. to 6.5 +/- 1.9 microgram/100 ml at 8 P.M. Sleeplessness for 24 or 48 hr under the conditions of this experiment failed to affect diurnal rhythms in aminopyrine metabolism or plasma 11-OHCS concentrations.


Spontaneous activities of female versus male newborns. This study explored potential sex differences in all activity spontaneously emitted by newborns. 2 groups of full-term infants (14 uncircumcised males, 15 females), equally distributed along several physical and demographic variables, served as subjects. Each newborn was observed for a total of 8 hours during the 2 days following birth. Observational procedures were stringently controlled and interobserver reliability ranged from .84 to .98. Results showed higher levels of wakefulness, facial grimacing, and low-intensity motor activity for males. Of marginal significance, both low-intensity and high-intensity oral activity was higher for males. In addition, a marginally significant interaction effect (sex X interfeeding interval X day) was found for high-intensity oral activity (crying). Independent of sex, high-intensity activity increased in the evening and over an interfeeding period, and low-intensity oral activity increased over days. The pattern of results indicates that even uncircumcised males may be slightly more irritable than females, and thus more active. Potential implications of these findings are considered.


[Idiopathic thrombocytopenic purpura in Hodgkin's disease--an early sign of a recurrence (author's transl)]. Idiopathic thrombocytopenic purpura (ITP) in the course of Hodgkin's disease is generally judged to be a paraneoplastic immune phenomenon. This concept finds support in the observation of a patient who developed simultaneously a recurrence of ITP and a protein anomaly in the form of an alpha-chain protein. Combined chemotherapy brought about remission of the Hodgkin's disease at the same time as normalisation of the platelet values and disappearance of the alpha-chain protein. There is no necessary correlation between tumour activity and ITP. But if ITP is observed, the most effective means would seem to be rigorous treatment of the Hodgkin's disease.


Effect of conformation of hCG-beta on generation of hCG-specific antibody. Most antisera generated to isolated highly purified beta subunits of human glycoprotein hormones are not sufficiently sensitive to detect physiologic blood levels of the native hormone. In the dissociated state, beta subunits assume a conformation different from that in the native hormone. Since antisera to alpha subunits have essentially no cross-reactivity between species, highly purified hCG-beta was combined with bTSH-alpha. That hybrid served as immunogen to assess whether sensitive, specific hCG antisera would more likely result than using hCG-beta alone. Of five animals immunized, three developed sufficiently sensitive and specific antisera. The results of these studies strongly suggests that human glycoprotein beta subunits combined with non-human alpha subunit are more likely to yield specific, sensitive antisera than when either isolated beta subunit or the native human glycoprotein hormone, containing common alpha determinants, serves as immunogen.


Virilizing effect of testosterone and its metabolites on the urovaginal septum of female fetal rats. A single injection of 50 microgram testosterone was given to fetal rats on day 17, 18, 19 or 20 of gestation. On day 21, the fetuses were removed from the mother under maternal ether anesthesia, and the length of the urovaginal septum was measured microscopically in female fetuses in order to assess the virilizing effect of testosterone. In fetuses treated with testosterone on day 17, the length of the urovaginal septum was comparable to that of oil-treated littermate controls. In fetuses treated on day 18, the length was significantly abridged compared with controls. In fetuses treated on day 19, the abridgment of the urovaginal septum was most marked. In fetuses treated on day 20, the length of the septum was again comparable to that of controls. The observations suggest that day 19 is the critical day for the virilizing effect of testosterone. Various amounts of testosterone and its metabolites including dihydrotestosterone, androstane-3 beta, 17beta-diol and androstane-3 alpha, 17beta-diol were injected into 19-day-old female fetuses, in order to test the dose relation to the virilizing effects of these steroids in terms of abridgment of the urovaginal septum. As a consequence, it was found that testosterone was the most effective for virilization.


The reactivity of a functional tyrosyl residue in carboxypeptidase B. Nitration of the cadmium enzyme. Cadmium-carboxypeptidase B was nitrated with tetranitromethane. The enzyme polymerized extensively during nitration. In the monomer nitrated Cd-carboxypeptidase B, 70% of the activity of Cd-carboxypeptidase B was retained. In order to identify the tyrosyl residues nitrated, the enzyme was digested with chymotrypsin and subtilisin and the nitrotyrosyl peptides were purified by affinity chromatography on antityrosyl-antibody-Sepharose conjugate followed by two-dimensional thin-layer chromatography. The major nitropeptides, representing 65% of the nitrotyrosyl label, were compatible with the segment of the sequence containing Tyr-240 and Tyr-259. Only 10% of the nitrotyrosyl label was found in the segment of Tyr-248. This indicates that the state of Tyr-248 in Cd-carboxypeptidase B differs from that in zinc-carboxypeptidase B where it shows chemical hyperreactivity due to its proximity to the metal ion.


Formation of 2-deoxyglucose-containing lipid-linked oligosaccharides. Interference with glycosylation of glycoproteins. Crude membrane preparations from chick embryo cells catalyse the formation of dolichyl-di-N-acetylchitobiosyl diphosphate [Dol-PP-(GlcNAc)2] from uridine diphosphate N-acetylglucosamine (UDP-GlcNAc). The formation of this glycolipid was stimulated by exogenous dolichyl phosphate and inhibited by tunicamycin. Adding GDP-mannose to the cell-free system containing Dol-PP-(GlcNAc)2 by preincubation led to the formation of a lipid-linked oligosaccharide, containing 8--9 sugar residues. The formation of lipid-linked oligosaccharides was inhibited by GDP-2-deoxy-D-glucose (GDP-dGlc): in this case Dol-PP-(Glc-NAc)2-dGlc accumulated. Subsequent additions of mannosyl residues to this trisaccharide-lipid to form lipid-linked oligosaccharides were not possible. Concomitantly the glycosylation of proteins was blocked. Partially inhibitory conditions were obtained by adding both GDP-dGlc and GDP-Man with an excess of GDP-dGlc. Glycosylation of proteins was observed but the glycopeptides did not contain 2-deoxyglucosyl residues. Also in these cases 2-deoxyglucose-containing glycolipids accumulated. The main glycolipid formed under these conditions was Dol-PP-(GlcNAc)2-Man-dGlc. Lipid-linked oligosaccharides containing 2-deoxyglucose were formed under these conditions, although in small amounts, but were not transferred to protein. So the molecular basis of the inhibitory action of 2-deoxyglucose on glycosylation of protein is the incorporation of 2-deoxyglucosyl residues during early phases of the biosynthesis of the lipid-linked oligosaccharides.


Monozygous twins discordant for amyotrophic lateral sclerosis. A pair of identical female twins were discordant for amyotrophic lateral sclerosis. The affected twin was not breast-fed; was bitten by a poisonous snake, and was operated on for struma. She had more infections than her sister but no fractures. Her plasma insulin response to glucose loading was also higher. The twins lived separated in their early childhood.


Effects of prostaglandin E2 applied locally on intravesical and intraurethral pressures in women. In 21 women undergoing simultaneous urethrocystometry because of dysuria, urge, and difficulties to empty the bladder, the effects of prostaglandin E2 (PGE2) applied locally in the bladder and urethra were investigated. In all 9 patients receiving PGE2 intravesically, the bladder pressure increased. Simultaneously, there was a signficant decrease in maximum urethral pressure (p less than 0.05), and in urethral closure pressure (p less than 0.005). Residual urine decreased in the 4 patients, in whom it exceeded 50 ml before administration. All 12 patients receiving PGE2 intraurethrally showed a decrease in maximum urethral pressure). There was a significant increase in bladder pressure (p less than 0.05); urethral closure pressure decreased in all patients. Bladder capacity decreased significantly (p less than 0.01). Residual urine exceeding 50 ml was found in 6 patients; it decreased in 4 after PGE2 administration. It is concluded that PGE2 applied intraurethrally can decrease the intraurethral pressure and increase the bladder pressure without side effects. By these actions, the drug might be useful for facilitating bladder emptying in patients with acute retention of urine.


"Behavioural despair" in rats and mice: strain differences and the effects of imipramine. Rats and mice when forced to swim in a restricted space will rapidly cease attempts to escape and become immobile. Previous experiments have shown that immobility was selectively reduced by antidepressant agents. The present experiments show that important differences exist between strains in both the amount of immobility observed and the effects of imipramine. Strain differences should therefore be taken into account in attempts to replicate results from one laboratory to another.


On the etiologic role of ureaplasma urealyticum (T-mycoplasma) infection in infertility. Two hundred consecutive infertility patients and sixty-seven controls subjects were studied for the incidence of infection with Ureaplasma urealyticum (T-mycoplasma). On the basis of a complete infertility investigation, the infertility patients were subdivided into those with explained infertility and those with unexplained infertility. Of the patients with unexplained infertility, 55% had a positive culture for T-mycoplasma as compared with a 32% incidence of positive cultures in the control population. The differences were statistically significant. The 6-month pregnancy rate following successful antibiotic treatment in patients with unexplained infertility was 42%. The 6-month pregnancy rate in a comparable group of patients with unexplained infertility, seen during a 3-year period prior to mycoplasma culture and treatment, was 32%. The difference in pregnancy rates between the two groups was not statistically significant. No correlation was found between a poor postcoital test and the presence of T-mycoplasma infection nor between T-mycoplasma infection and poor cervical mucus. The role of T-mycoplasma infection in infertility was neither proven nor disproven by this study.


Ureaplasma urealyticum (T-mycoplasma) in vaginal fluid and cervical mucus from fertile and infertile women. Ureaplasma urealyticum (T-mycoplasma) was isolated more frequently and in heavier growth from cervical mucus (49%) than from vaginal fluid (34%). It was isolated in 24% of vaginal fluid samples and in 35% of cervical mucus samples from fertile women, and in 29% of vaginal fluid samples and in 47% of cervical mucus samples from infertile women. The incidence of infection was high following abortion or total hysterectomy and during pregnancy or oral contraceptive use. T-mycoplasma was also isolated from the vaginal fluid and cervical mucus of a woman with tubo-ovarian abscess, but was not present in women with Trichomonas vaginalis infection. U. urealyticum did not alter the physiophysiologic characteristics of vaginal fluid and cervical mucus or the sperm penetration and sperm viability in cervical mucus. Treatment with tetracycline eradicated the organism in 88% of the infected women. Pregnancies were recorded during a 6-month follow-up in 1 of 19 infertile women who were treated with tetracycline.


Traumatic hypomenorrhea-amenorrhea (Asherman's syndrome). Traumatic damage to the endometrium (corporeal adhesions) and/or to the cervical internal os (cervical stenosis or atresia) may cause hypomenorrhea-amenorrhea, the only sign of Asherman's syndrome. We demonstrated a close correlation between severity of menstrual insufficiency and extent of corporeal adhesions. Minor adhesions in the lower part of the uterine cavity, however, may cause severe hypomenorrhea, frequently complicated by dysmenorrhea. Secondary amenorrhea may be due either to complete obliteration of the uterine cavity or to stenosis or atresia of the internal os, as proved by radiologic studies. When amenorrhea is due to stenosis or atresia of the internal os, the ovarian cycle continues but the endometrium becomes refractory to horomonal stimuli, and hematometra does not occur. Simple cervical dilatation restores menstruation within 4 to 5 weeks. The menstrual insufficiency of Asherman's syndrome, therefore, may be explained by two pathophysiology mechanisms: (1) reduction of the endometrial bleeding area and (2) trophic changes and unresponsiveness of the endometrium, perhaps caused by a visceral reflex originating in the area of the internal os.


Cytochalasin B stabilises the sub-cortical actin bundles of Chara against a solution of low ionic strength. The properties of the sub-cortical actin bundles in the perfused Chara cell model are altered by concentrations of cytochalasin B (CB) which inhibit streaming. This is demonstrated by treating the bundles with intracellularly introduced CB after first using ATP to strip away their associated motile organelles. Such CB-treated bundles are resistant to a solution of low ionic strength which, as judged by light and electron microscopy, extracts untreated bundles. It is therefore concluded that a site of action for CB lies within the actin bundles and that, while the alteration detected may not itself be of direct physiological significance, it may nonetheless indicate the site at which CB inhibits streaming in both the cell model and the intact cell.


The changes in ultrastructure during fertilization of the colourless flagellate Polytoma papillatum with special reference to the configural changes of their mitochondria. Changes in the morphology of the mitochondrial inventory (= chondriome), the nucleus and the flagellar apparatus during the generative (sexual) life cycle of Polytoma papillatum were examined by means of the serial sectioning technique. At the onset of copulation gametes do not differ obviously from interphase cells of the vegetative (asexual) life cycle, in that, both primarily contain one basket-shaped mitochondrion. The quadriflagellate and binucleate zygote exhibits a chondriome which consists of one large highly reticulated basket at the periphery of the zygote and 33 smaller mitochondrial units. Therefore, the basket clearly results from fusion of the two gamete chondriomes. The smaller mitochondrial fragments are either spherical to ovoid or elongated and poorly branched; they tend to occupy more central regions of the zygote. After karyogamy the mitochondrial basket disintegrates into several fragments of various shapes and sizes. Most of the mitochondrial fragments are located at the periphery. At the onset of karyogamy the nuclei and the flagellar apparatuses do not differ significantly from those of the gametes and vegetative interphase cells. The diploid nucleus, however, is characterized by: 1. many spherical bodies (diameter: ca. 200 to 600 nm) which are found both in the nucleoplasm and in the nucleolus. The major part of these bodies consists of material whose ultrastructure resembles that of the "pars fibrosa" in the nucleolus; 2. three deep invaginations of the nuclear membrane, two of which extend to the nucleolus; 3. an increase of nucleoplasm-filled cavities in the nucleolar "pars granulosa". The four flagella are considerably shortened; the basal bodies bound to the flagella have lost their striated connection and the roots have nearly completely disappeared. The results are compared with those obtained from investigations in Chlamydomonas; their significance in extranuclear genetics and in the systematics of Volvocales is discussed.


Structure, synthesis and orientation of microfibrils. V. On the recovery of Oocystis solitaria from microtubule inhibitor treatment. Depending on the type of the inhibitor and its concentration one can experimentally induce two forms of aberrant microfibril orientations in O. solitaria cell walls through microtubule inhibitor application. The first form, designated "Intermediate", is characterized by the presence of cortical microtubules together with a spiral arrangement of microfibrils. The second form, designated "Parrallel", shows a wall with bundles of parallel oriented microfibrils without cortical microtubules. Taking colchicine as an example for a microtubule-inhibitor the "Parallel" form may be obtained with 10mM and the "Intermediate" with 5 to 1 mM solutions. Some microtubule-inhibitors such as methylbenzimidazole-2yl-carbamate (MBC) produce the "intermediate" form only. The recovery of normal microfibril orientation after inhibitor treatment is dependent on three factors: a) the developmental stage--young autospores just beginning to synthesize a wall are absolutely necessary; b) the application of inhibitors with the lowest effective concentration for c) the shortest possible time. Minimal concentrations for obtaining a "Full" effect range from 10 mM for colchicine to 1 micrometer for amiprophosmethyl (APM) with incubation periods from 3 to 9 hours. The return to the normal microfibril orientation has been achieved in all cases except after podophyllotoxin treatment. Since APM has been claimed to act selectively on tubulin synthesis in Chlamydomonas it was decided to compare the effects of this compound with cycloheximide (10 microgram/ml) on the recovery of microfibril orientation after colchicine treatment. In both cases no orientation recovery is possible although in the case of cycloheximide, synthesis of cellulose is drastically inhibited. This cycloheximide inhibition is fully reversible. During cycloheximide, but not APM, inhibition cortical microtubules return; however, due to the inhibition of cellulose synthesis itself, they cannot exert their orienting influence.


Morphological and autoradiographical characterization of membrane-like layers surrounding cultured fibroblasts. Two membrane-like layers of approximately 7 nM thickness were observed in ultrathin cross-sections of flat-embedded cultured fibroblasts. One "membrane", underlying the cell-monolayer, possibly established contacts between cells and substratum. The other "membrane" covered the whole cell culture. These "membranes" were at least partially synthesized by the cells themselves, as was shown by high resolution autoradiography. With regard to our biochemical findings, the autoradiographical results showed that components of these "membranes" consisted of sulfated proteoglycans and may be the morphological equivalent to the biochemically well-defined cell surface associated material, the so-called pericellular pool of glycosaminoglycans.


Hypovitaminemia A in idiopathic hemochromatosis and hepatic cirrhosis. Role of retinol-binding protein and zinc. Serum levels of vitamin A, its specific carrier protein retinol-binding protein (RBP), and zinc were determined in 34 cases of idiopathic hemochromatosis, 33 cases of alcoholic cirrhosis, 10 cases of non-alcoholic cirrhosis, and in 35 normal controls. In both alcoholic and non-alcoholic cirrhosis, vitamin A and RBP levels were very significantly reduced, whereas a significantly low zinc was observed only in the alcoholic cirrhosis group. In idiopathic hemochromatosis, vitamin A values were significantly lower compared to normals, whereas serum RBP levels were normal and serum zinc was very close to that of the controls. A significant correlation was found between vitamin A and RBP levels in the entire group of 112 patients. These results, (1) in alcoholic and non-alcoholic cirrhosis, confirm a dramatic vitamin A deficiency and the major role played by decreased RBP, but tend to deemphasize the possible role of zinc deficiency; (2) in idiopathic hemochromatosis, affirm a significant serum vitamin A deficiency supposedly by a different mechanism from that of alcoholic cirrhosis since in idiopathic hemochromatosis plasma RBP levels are normal. The role of this vitamin A disorder should be considered in the interpretation of clinical signs of idiopathic hemochromatosis such as ichthyosis and visual disorders.


[Echocardiography in obstructive hypertrophic myocardiopathy. Proposed index of severity of the obstruction]. An obstruction index to estimate the left ventricle outflow gradient from the echocardiogram in patients with obstructive cardiomyopathies is proposed. It is based on the measurements of the mean distance between septum and anterior mtral valve leaflet, the total time of narrowing and the time of minimal and maximal narrowing. In the first group of patients studied, it seems to correlate fairely with the severity of clinical picture and with the gradient found at catheterization.


Induction of granulomas in mice by Crohn's disease tissue. Normal and immunodeficient CBA and A2G strain mice were inoculated with crude (100 to 20 to 30 micrometer), cell-free (0.2 micrometer) filtrates of Crohn's or noninflammatory bowel disease tissue homogenates, which were either fresh or frozen to -70 degree C. Mice of each strain developed epithelioid and giant cell granulomas both locally at the site of injection and systemically in response to crude and cell-free filtrates of Crohn's tissues. Control mice did not develop such changes. The granulomas evolved slowly, predominantly between 9 and 27 months. The granuloma-inciting agent has been shown to be present in ileum, colon, and mesenteric lymph nodes of patients with Crohn's disease and it withstands freezing to -70 degree C. The use of Crohn's tissues common to this study and one in rabbits previously reported, suggests that the induction of granulomas by this agent is not strain- or species-specific, and is independent of the immune status of CBA mice.


Comparison of an intragastric method of estimating acid output with the pentagastrin test in normal and duodenal ulcer subjects. Using Fordtran's technique but substituting the meat extract Oxo for the steak meal we investigated gastric acid secretion in eight control subjects and nine patients with chronic duodenal ulcer. Intragastric titration was performed using a double lumen tube measuring the pH in the stomach every three minutes and adjusting it to 5.5 throughout the test by infusing 0.3-M sodium bicarbonate. On a separate day a pentagastrin test was performed using a conventional gastric aspiration technique. In the eight control subjects the mean acid output after pentagastrin was 13.7 +/- 2.1 (SEM) mmol/h, whereas the mean hourly acid output measured by intragastric titration was 20.1 +/- 3.1. The greater response to Oxo than to pentagastrin in the controls (deltaAO = + 46%) was significant (P less than 0.01). This is in contrast with our duodenal ulcer patients whose mean hourly acid outputs were 22.7 +/- 4.4 and 23.0 +/- 4.4 mmol/h in response to pentagastrin and Oxo respectively (r = 0.95). The findings, while clearly at variance with those of Fordtran and Walsh (1973), are more in keeping with the concept of increased endogenous secretory drive in duodenal ulcer patients compared to normal subjects.


Pharmacokinetics of pancreatic polypeptide in man. Pure bovine pancreatic polypeptide (PP) was infused into 23 healthy subjects at doses of 1, 3, and 5 pmol/kg/min over 60 minutes and plasma PP was measured by radioimmunoassay. During the infusions mean plasma levels of 203 +/- 34, 575 +/- 73, and 930 +/- 48 pmol/l respectively were achieved. Mean disappearance half time on stopping the infusion was 6.9 +/- 0.3 min (mean +/- SEM). The metabolic clearance rate was 5.1 +/- 0.2 ml/kg/min (mean +/- SEM) and the apparent volume of distribution was calculated to be 51 +/- 3 ml/kg (mean +/- SEM). This study provides for the first time pharmacokinetic data for PP in man.


A comparison between diabetics receiving a high or low daily insulin dosage. Two groups of 8 diabetics, similar in many aspects, who required high (greater than or equal to 128 units) or low (less than or equal to 32 units) total daily insulin dosage were compared during a 24 hour period. C-peptide secretion was detected in all but one of the low dose group, but only in two of the high dose group. No significant differences in serum free and total insulin concentrations were detected and the patterns of blood metabolites were also similar, apart from higher blood glycerol concentrations in the low dose group. No significant differences in the binding characteristics of the antiinsulin antibodies were detected.


[Limitations of mass transfusion].

[Limitations of mass transfusion]. As a result of blood conservation there is nearly no limit to transfusion therapy. This is shown by reviews of the University Clinic of Hamburg-Eppendorf. A still existent limitation in patients with irregular antibodies requires a supraregional cooperation the possibilities of which are discussed. In intensive care medicine an especially critical indication is necessary. Blood transfusion in hopeless cases (ut aliquid fiere) is as unjustifiable as artificial respiration is in patients having an EEG 0-line.


[Current status of granulocyte separation and transfusion]. Granulocyte substitution therapy is of definitive clinical value when correctly indicated and consequently performed. Comparing the different techniques for leucocyte separation, the combination of continuous flow centrifugation (CFC) and filtration (FL) permits the highest yield of granulocytes. As yet, all available methods imply a rather high degree of inconvenience and a considerable rate of side effects for the (normal) donor which cannot easily be justified from a medical point of view. On the other hand, CFC and especially FL have a great potential for further improvement. Hence, higher yields and less donor affection are expectable. Granulocyte procurement is a great challenge to existing transfusion centres requiring their future engagement and expansion in this new and important field.


Effect of magnesium deficiency and food restriction on the immune response in young mice. Similar batches of five-week-old C57 Bl/6 mice were given either a magnesium-deficient diet (4mg Mg/100 g), or a control diet (40 mg/100 g). Control diet intake was either ad libitum or reduced. After immunization with SRBC (sheep red blood cells), the immune response was studied by estimating the number of spleen AFC (antibody-forming cells) capable of lysing SRBC, and by a cytoadhesion test to determine the number of RFC (rosette forming cells). Limitation of the control diet slowed the growth rate in mice. Whenever food intake was reduced from 4g/day to 2.9 or 2.6g/day, the AFC response intensified but the RFC response remained similar. Food limitation might therefore mainly affect immature IgM producing cells with a high dividing rate. Magnesium deficiency produced a drastic fall in the primary and secondary immune responses, as measured by the number of spleen AFC. The number RFC was also much lower in the spleen of deficient animals. Consequently, the spleen immune system is deeply affected by this deficiency.


Receptor protein for dihydrotestosterone in nuclei of the female prostate of Praomys (Mastomys) natalensis. An investigation was carried out to identify and characterize the binding of androgens within the nuclei of the prostate from female Praomys (Mastomys) natalensis. After in vivo and in vitro labeling with tritiated androgens, the tissue was subjected to cell fractionation and the nuclear fraction was purified. Gel exclusion chromatography and sucrose density gradient centrifugation of the nuclear extract showed the presence of a steroid receptor complex having a sedimentation coefficient of 3 S. Subsequent analysis of the bound radioactivity revealed that more than 90 per cent of the recovered tritiated steroid was dihydrotestosterone. This binding was found to be temperature dependent and could not be shown in liver tissue. The behavior of this nuclear steroid receptor complex is compared with that of the male Mastomys and the rat. It is suggested that the female Mastomys could be used as an experimental model for studying hormone effects on the prostate.


Bladder and urethral responses to isolated sacral motor root stimulation. After complete intradural lumbosacral rhizotomy and cord transection in 40 cats, parasympathetic discharge was induced by electrical stimulation of the second and third sacral roots. Isolation of the stimulus to the preganglionic motor fiber resulted in bladder contraction and reciprocal urethral pressure loss. The threshold for the urethral response was lower than that of the bladder and the urethral response findings strongly suggest that urethral adrenoreceptor activity can be modified peripherally by a primary parasympathetic stimulus.


[Consulting office tissue banks (author's transl)]. The technicalities of obtaining graft tissues and the methods of preserving homograft ossicles and cartilage are described. The chemical and biological problems of tissue banks are discussed practically in relation to published experiences of others. The techniques for use of homograft ossicles in tympanoplasty surgery are described. The comparative ease of preparation and preservation of homograft ossicles and hyaline cartilage justifies the establishment of tissue banks in the practice of the private otologist.


Ozone depletion: implications for the veterinarian. Man has inadvertently modified the stratosphere. There is a good possibility that the ozone layer is being depleted by the use of jet aircraft (SST), chlorofluoromethane propellants, and nitrogen fertilizers. Under unpolluted conditions, the production of ozone equals its destruction. By man's intervention, however, the destruction may exceed the production. The potential outcome is increased intensity of solar ultraviolet (280-400 nm) radiation and penetration to the earth's surface of previously absorbed wavelengths below about 280 nm. The increased ultraviolet radiation would increase the likelihood of skin cancer in man and ocular squamous cell carcinoma in cattle. The climate also might be modified, possibly in an undesirable way.


Fetal alcohol damage and schizophrenia.

Fetal alcohol damage and schizophrenia. The case of a 15 year old girl with fetal alcohol damage and schizophrenia is presented. Because fetal exposure to alcohol may produce brain damage, itself associated with increased vulnerability for schizophrenia, and because chronic exposure to alcohol may lead to a symptomatic schizophrenic illness, prospective study of children with fetal alcohol damage is needed to determine whether they are at greater risk for the later development of schizophrenia.


Thermal adaptation in yeast: obligate psychrophiles are obligate aerobes, and obligate thermophiles are facultative anaerobes. The obligate psychrophilic yeasts Torulopsis psychrophila, T. austromarina, Leucosporidium frigidum, L. gelidum, and L. nivalis were obligate aerobes and were unable to grow anaerobically. In contrast, the obligate thermophilic yeasts T. bovina, T. pintolopesii, Candida slooffii, and Saccharomyces telluris were facultative anaerobes.


Enzymic racemization of allantoin.

Enzymic racemization of allantoin. Allantoin racemase was isolated from cells of Candida utilis, and purified by chromatography on columns of DEAE-cellulose and Sephadex G-100. Using this purified enzyme, the racemization of allantoin in deuterium oxide was investigated. Polarimetric and PMR spectroscopic analyses showed that racemization of allantoin by the enzyme proceeded in parrallel with release of the hydrogen atom (5-H) attached to the asymmetric carbon (C-5) of allantoin. Non-enzymic racemization of allantoin, which was examined for comparison, however, was accompanied by much less or almost no release of allantoin 5-H. This indicates that the mechanism of racemization by the enzyme differs from that of non-enzymic racemization.


Failure of the usual anti-Ig antibody method for quantitative measurement of low affinity antibodies. The usual anti-Ig antibody method, consisting of the precipitation of soluble antigen-antibody complexes by heterologous anti-Ig antibody, was applied for quantitative estimation of guinea pig IgG2 anti-ovalbumin and anti-2,4-dinitrophenyl (DNP) antibodies by measuring the maximum amounts of antibody-bound antigens. However, the amounts of antibodies estimated were less than those obtained by other methods: the precipitin reaction, the precipitation of antigen-antibody complexes with 50% saturated ammonium sulfate, and equilibrium dialysis. In particular, the anti-Ig antibody method greatly underestimated the amount of anti-DNP antibody with low affinity for epsilon-DNP-L-lysine. Thus, it was concluded that partial dissociation of the antigen-antibody complexes occurring upon precipitation with anti-Ig antibody made the anti-Ig antibody method unsuitable for quantitative determination of antibodies.


Resonance Raman spectra of riboflavin and its derivatives in the bound state with egg riboflavin binding proteins. The resonance Raman spectra of riboflavin (RF) and its derivatives, including 3-deuterated (3-D RF), 3-methyl (3-CH3 RF), 3-carboxymethyl (3-CH2COOH RF), and 7,8-dichlororiboflavins (7,8-Cl RF), in H2O and D2O were observed in the 700-1700 cm-1 region. The fluorescence problem of riboflavin was overcome by complex formation of riboflavin with riboflavin binding proteins. The observed frequencies of Raman lines of RF are in good agreement with those of glucose oxidase obtained by Spiro et al. by the resonance CARS method, although the present spectral range is extended to much lower frequency with a higher signal-to-noise ratio than that for glucose oxidase. The observed Raman lines were assigned to the individual ring modes of isoalloxazine on the basis of the Raman spectra of appropriate model compounds such as uracil, pyrazine, and o-xylene. The 1253 cm-1 line of RF was shifted to ca. 1300 cm-1 for 3-D RF, 3-CH3 RF, and 3-CH2COOH RF, and accordingly can be assigned to the CN stretching mode of Ring III. The 1632 cm-1 line of RF was shifted for 7,8-Cl RF and was assigned to a Ring I mode. No Raman line mainly due to C = O stretching mode was observed in the present resonance Raman spectra.


Effects of albumin on the phagocytosis of polystyrene spherules by rabbit polymorphonuclear leucocytes. Polystyrene latex spherules are rapidly phagocytized by polymorphonuclear leucocytes. Surface modifications of the latex spherules can lead to an inhibition of phagocytosis, and the purpose of this study was to analyse in particular inhibition by bovine and human serum albumin. It can be shown that albumin is bound much more strongly by the PSL particle surface than by the cell surface. Furthermore, a correlation was established between the extent to which the particle surface is covered and the extent of phagocytosis inhibition. In the binding of albumin to the particle surface ionic as well as hydrophobic interactions play a role. As a result, the particle acquires a negatively charged hydrophilic coating which effectively diminishes the number of effective cell-particle collisions leading to engulfment.


Genetic characterization of hydroxyurea-resistance in Chinese hamster ovary cells. Hydroxyurea is an excellent selective agent for obtaining drug-resistant mutants. At a frequency of approximately 1 X 10(-5) it was possible to select, in a single step, colonies that exhibited significant resistance to the cytotoxic effects of the drug. These hydroxyurea-resistant cell lines maintained their resistant phenotype after extensive cultivation in the absence of the drug. Reconstruction experiments indicated that the expression of hydroxyurea-resistance and the frequency of drug-resistant colonies was independent of cell densities up to 5 X 10(5) cells per 100-mm selection plate. Luria-Delbrück fluctuation analyses indicated that the appearance of hydroxyurea-resistant cells in wild type populations occurred spontaneously and at a rate of 4.8 X 10(-6) per cell per generation in the presence of 0.33 mM drug. Studies with the mutagen, ethyl methane sulfonate indicated that it was capable of increasing the frequency of hydroxyurea-resistant cells by a factor of approximately 10. Also, cell-cell hybridization experiments showed that hydroxyurea-resistance behaves as a dominant or codominant trait and that hydroxyurea-resistance was a useful new genetic marker for selection of somatic cell hybrids. Furthermore, similar to many other drug-resistant cell lines hydroxyurea-resistant cells were found to exhibit an altered sensitivity to a number of non-selective agents (guanazole, N-carbamoyloxyurea, formamidoxime, and hydroxyurethane). Except for guanazole these compounds are structurally very similar to hydroxyurea and may be expected to have similar modes of action. The results presented in this paper support the view that hydroxyurea-resistance is expressed as a normal genetic trait and is a useful genetic marker for somatic cell genetic studies.


Ribonucleotide reductase from wild type and hydroxyurea-resistant chinese hamster ovary cells. The kinetic properties of partially purified ribonucleotide reductase from Chinese hamster ovary cells have been investigated. Double reciprocal plots of velocity against substrate concentration were found to be linear for three the substrates tested, and yielded apparent Km values of 0.12 mM for CDP, 0.14 mM for ADP and 0.026 mM for GDP. Hydroxyurea, a potent inhibitor of ribonucleotide reduction, was tested against varying concentrations of ribonucleotide substrates and inhibited the enzyme activity in an uncompetitive fashion. Intercept replots were linear and exhibited Ki values for hydroxyurea of 0.08 mM for CDP reduction, 0.13 mM for ADP reduction and 0.07 mM for GDP reduction. Guanazole, another inhibitor of ribonucleotide reductase, interacted with the enzyme in a similar manner to hydroxyurea showing an uncompetitive pattern of inhibition with CDP reduction and yielding a Ki value of 0.57 mM. Partially purified ribonucleotide reductase from hydroxyurea-resistant cells was compared to enzyme activity from wild type cells. Significant differences were observed in the hydroxyurea Ki values with the three ribonucleotide substrates that were tested. Also, CDP reductase activity from the drug-resistant cells yielded a significantly higher Ki value for guanazole inhibition than the wild type activity. The properties of partially purified ribonucleotide reductase from a somatic cell hybrid constructed from wild type and hydroxyurea-resistant cells was also examined. The Ki value for hydroxyurea inhibition of CDP reductase was intermediate between the Ki values of the parental lines and indicated a codominant expression of hydroxyurea-resistance at the enzyme level. The most logical explanation for these results is that the mutant cells contain a structurally altered ribonucleotide reductase whose activity is less sensitive to inhibition by hydroxyurea or guanazole.


Dietary lipid metabolism in lactating dairy cows. Effects of feeding an oil seed supplement treated with formalin upon lipid patterns of blood and synthesis of milk fat were evaluated. Percentages and yields of fatty acids of milk fat with chain lengths between 6 and 16 carbons were decreased while percentages and yields of stearate and linoleate were increased when the lipid supplement was fed. Calculations in cows fed control and supplement, 60% and 80%, respectively, of fatty acids of milk were derived from lipids of blood were supported by arterial-venous differences. Comparisons of the fatty acid compositions of triacylglycerol of plasma and milk fat suggested that triacylglycerol may not be the sole source of linoleate transferred from blood to milk fat. A preliminary evaluation of supplement effects upon lipoprotein patterns of serum indicated two peaks in the low density lipoprotein class and that the increase in total cholesterol of blood caused by feeding lipid supplement is due to increases in cholesterol content of the low density and high density lipoprotein classes.


Metabolic control of the circulation. Effects of acetate and pyruvate. Chloralose-anesthetized dogs were infused intravenously with either Tris-acetate or Tris-pyruvate at 0.0375, 0.075, and 0.15 mmol/kg per min successively, each for 20 min. Acetate infusion increased cardiac output, left ventricular dP/dt and dP/dt/P, and coronary blood flow, while pyruvate infusion did not. Infusions of either substance increased arterial blood and skeletal muscle concentrations of citrate and malate, but only acetate infusion increased the tissue AMP content and decreased the ATP:AMP ratio. The increase in cardiac output produced by acetate was accompanied by an increase in total body oxygen consumption and a decrease in the difference between arterial and mixed venous blood oxygen. Myocardial oxygen consumption increased during acetate infusion, but the decrease in myocardial oxygen extraction and the increase in coronary sinus blood oxygen saturation suggest that an active coronary vasodilation which was not a result of the increased cardiac work, occurred. The concentration of hypoxanthine in the coronary sinus and the content of myocardial adenosine increased, which suggests that the increase in coronary blood flow was caused by the vasodilator action of adenosine released from the myocardium, and that adenosine production is not necessarily tied to PO(2). These systemic and coronary hemodynamic changes also occurred when acetate (0.075 mmol/kg per min) was infused into conscious dogs. Acetate infusion also increased blood flow to the gastrointestinal tract, kidneys, intercostal muscle, and diaphragm. These changes were not affected by propranolol pretreatment, but were abolished by pretreatment with fluoroacetate which reduced acetate oxidation. These results suggest that the circulatory stimulation produced by acetate was not caused by increases in tricarboxylic acid cycle intermediates. Instead, it was probably related to the increased cleavage of ATP to AMP that accompanies activation of acetate to acetyl CoA, and was not mediated via beta-adrenergic receptors. It is speculated that hemodynamic changes may occur in patients who undergo hemodialysis with acetate-containing dialysate. Hemodynamic changes of ethanol may also be brought about by acetate, which is one of the intermediates that accumulates during ethanol metabolism.


Influence of dietary protein concentration on milk production by dairy cattle during early lactation. In a 3 X 2 factorial experiment 75 Holstein cows in first, second, or third lactation were fed rations containing either 12.2% or 16.2% crude protein in total ration dry matter. On the average, 26% of dry matter intake was from corn silage, 22% from alfalfa-grass hay, and 52% from a grain mix. Protein was controlled by feeding a 13.7% crude protein grain mix with 1.4% urea for the 12% ration and a 19.8% crude protein grain mix with natural protein for the 16% ration. Average daily milk production (kg/day) for wk 2 through 12 of lactation for 12% and 16% rations by lactations were: first, 21.6 and 21.9; second, 25.7 and 31.5; and third, 27.5 and 34.0. Dry matter intakes by lactations were .42, 1.18, and 2.05 kg/day higher for cows fed the high protein compared to low protein rations. Milk composition was not influenced by protein treatment. The markedly different response to protein supplementation in milk production between heifers in first lactation and more mature cows is unexplained.


Abomasum emptying in young dairy calves fed milk replacers containing animal or vegetable fats. Young Holstein male calves, fitted with re-entrant duodenal fistulae, were fed 25% fat milk replacers containing either butter oil, lard, or corn oil in a 3 X 3 Latin square experiment. Proteins were supplied by skim milk powder. Patterns of changes with time after feeding were observed for the composition and pH of duodenal digesta, recovery rates of fresh and dry matter, rates of abomasum emptying for fresh and dry matter, gastric proteolysis, and diarrhea. The composition and pH of duodenal digesta changed markedly with time after feeding, but the only notable differences between milk replacers were for contents of protein nitrogen and total nitrogen. Recovery rates of fresh matter were higher than 100%, presumably due to salivary and gastric secretions. Dry matter was recovered at a rate averaging 100% suggesting that the method used to measure the flow rate and sample duodenal digesta was adequate. The highest flow rates of fresh matter were during the first 3 h after feeding. Differences were marked among the flow rates of constituents of dry matter. Gastric proteolysis was not influenced by the nature of dietary fats. Fecal dry matter, a measure of the severity of diarrhea, was markedly lower for the milk replacer filled with corn oil than for the other two milk replacers.


Hemangiosarcoma subsequent to radiotherapy for a hemangioma in infancy. A case is presented of a 31-year-old white man who developed a hemangiosarcoma in the very place where 30 years ago a hemangioma had been treated by radiotherapy. The rarity of such an occurrence is attested to by the fact that only two similar cases were found in the English medical literature.


Ultrastructural localization of acetylcholinesterase in cultured cells. III. DFP treated embryo muscle. Brief treatment with 10(-4)M diisopropylfluorophosphate (DEP) irreversibly inactivates acetylcholinesterase (E.C.3.1.1.7; acetylcholine hydrolase) (AChE) activity in 10 day old chick embryonic muscle cultures. Electron microscopic cytochemistry was employed to follow the distribution of new AChE during recovery from DEP treatment. In normal 10 day cultures of embryo pectoralis muscles AChE is localized in the nuclear envelope, perinuclear sarcoplasm, sarcotubular system, subsurface vesicles and bound outside the cells. Immediately after DFP treatment AChE activity is absent in large myotubes. Within 15 min, activity is randomly present in small amounts in the sarcotubular system and nuclear envelope. There is a dramatic increase in activitv in the nuclear envelope during the 1st hr of recovery, and connections between the nuclear envelope and sarcotubular system are often seen. The next few hr of recovery show increased AChE activity. By 4 hr activity approaches that of controls. Six to 8 hr after treatment, AChE activity can be detected spectrophotometrically in the medium and can be seen bound outside the cells with the electron microscope. The spatial and temporal patterns of AChE activity demonstrate that the recovery of AChE and its mobilization and release from DFP-treated cells are not governed solely by the levels attained by the enzyme in the cultured embryo muscle.


Thrombocytopenia in polycythemic mice.

Thrombocytopenia in polycythemic mice. Significant and sustained thrombocytopenia in mice maintained in a polycythemic state with weekly injections of washed packed RBCs prompted the examination of the relationship between the observed thrombocytopenia and the polycythemic state. Platelet production was determined by measurement of 75SeM incorporation into platelets. Platelet survival was studied with 51Cr-labeled platelets. Platelet sequestration was assessed by measuring the trapped platelet 51Cr-bound radioactivity in the spleens of these animals. Blood volume was calculated by measuring the dilution of injected 59Fe-tagged RBCs. These studies failed to reveal decreased platelet production, a shortening of the platelet survival, or significant splenic pooling as factors contributing to the observed thrombocytopenia. The blood volumes in the polycythemic group of animals were 8.9% +/- 0.15 (1 S.D.) body weight vs. 6.4% +/- 0.69 in the normals (p less than 0.001). Contrary to what has been reported for hypoxia-induced polycythemia, our results indicate that transfusion-induced polycythemia does not cause thrombocytopenia by decreasing platelet production. We suggest instead that the observed thrombocytopenia is mainly due to dilution of the circulating platelet mass by the expanded blood volume.


The extracorporeal bile duct: a new model for determination of bile flow and bile composition in the intact rat. A new model is described which allows measurement of bile flow and sampling of bile in the intact rat with a physiologically functioning sphincter of Oddi. A number of metabolic parameters have been followed to show that animals with such an "extracorporeal bile duct" (EBD) behave as intact controls. Especially, there was no difference in the increase in body weight or hepatic fatty acid and cholesterol synthesis between EBD animals and intact controls. The amount of bile salts circulating through the biliary tract amounted to 30.5+/-1.5mumol . 100 g body wt-1 . hr-1, when diurnal variations were averaged. Animals adapted to a restricted feeding regimen showed a significant increase of bile flow and of biliary bile salt and cholesterol excretion during feeding (10AM-3 PM); these parameters reached their maximum 3 hours after onset of food intake.


Lipase hydrolysis of mammalian long-chain 1,2-alkanediol diesters. Nonrandom distribution of fatty acids. Long-chain 1,2-alkanediol diesters were isolated from the total surface lipids of golden Syrian hamsters and Swiss albino mice. Hydrolysis of the diol diester waxes with exocellular lipase from Rhizopus arrhizus delemar or with purified porcine pancreatic lipase produced free fatty acids and 2-acyl diols in about 60--80% yield. Nonrandom distribution of the constituent fatty acids at positions 1 and 2 of the alkanediols was observed. In the diester waxes from the hamster, both straight-chain and branched-chain fatty acids of 14 to 20 carbon atoms predominated at position 1 and those of 22 to 26 carbon atoms at position 2. In contrast, the diester waxes of the mouse contained mainly fatty acids of less than 19 carbon atoms, both saturated and monounsaturated, at position 2 and those of greater chain length (20 to 24 carbon atoms) at position 1. The results of the lipase hydrolysis were confirmed by degradation of the diester waxes with Grignard reagent.


The preparative isolation of lecithin.

The preparative isolation of lecithin. Lecithin can be prepared on a relatively large scale, free of colored impurities, by a simple two-column procedure. Commercial crude egg lecithin is partially purified by a single-step passage through an alumina column. It is then purified by a two-step passage through a prepacked, commercial silica gel column. The lecithin is prepared in solvent-free form for weighing by lyophilization from cyclohexane. Toxic solvents (chloroform and methanol) are avoided by the use of ethanol, isopropanol, hexane, and water. The elutions are easily monitored by a flow cell in an ordinary spectrophotometer set at 215 nm. Study of the column parameters has made it possible to use heavy loads with a relatively small column and minimal solvent.


[Causes, diagnosis and treatment of uterine perforations by intrauterine devices]. Perforations of the uterus by intra-uterine devices (I.U.D.) are still at present serious complications. Their incidence is very varied and diverse because they are often asymptomatic. Pregnancy of a frequent circumstance under which they are discovered, as well as the more general way, which is the disappearance of the strings, which should always give rise to the suspicion that perforation has happened. There are two types to be distinguished from one another: Immediate perforation due to a faulty technique in insertion, secondary perforation due to gradual wearing down of the uterine wall. The factors predisposing to the occurrence of these accidents are numerous but notable among them are the date in the cycle when the insertion was done and above all the time in relationship to the last pregnancy, parity, the position of the uterus and the experience of the operator. Diagnosis of perforation is not always easy and may require not only radiological investigation without preliminary preparation but also hysterography, ultrasound examination and hysteroscopy. Withdrawal of a device that has perforated the uterus is advisable even if its migration has not given rise to any clinical symptoms. The technique for removing it varies often from case to cause between laparoscopy, laparotomy and culdotomy. Prevention of perforations should not only take into account the contra-indications to insertion but also knowledge of favourable factors, of a rigorous technique for insertion as well as clinical supervision in order to screen for secondary perforations.


Echocardiographic left atrial enlargement. Comparison of vectorcardiogram and electrocardiogram for detection. Standard electrocardiograms (ECG) and Frank vectorcardiograms (VCG) were obtained in 43 consecutive patients in sinus rhythm who had echocardiographic evidence of left atrial enlargement (left atrial internal dimension greater than 4.0 cm; x +/- 1SD = 4.7 +/- 0.5 cm).  High gain VCG P loop measurements for the study group were: maximal posterior magnitude, 0.11 +/- 0.03 mv; duration, 106 +/- 14 msec and ratio of maximal posterior to maximal anterior P vector magnitudes, 3.2 +/- 1.4. Thirty of 43 (70%) patients with echocardiographic determined left atrial enlargement had VCGs diagnostic of that condition. Utilizing New York Heart Association criteria for left atrial enlargement, 17 of 43 patients (40%) had ECGs which were diagnostic. Fifteen of 43 (35%) subjects manifested both ECG and VCG criteria for left atrial enlargement and only two patients had diagnostic ECGs and normal VCGs. It is concluded that analysis of high gain VCG P loops provides a 30% higher yield for the diagnosis of echocardiographically determined left atrial enlargement when compared with P wave examination on the standard ECG.


Effect of anaesthesia on ovarian follicular development and ovulation in the sheep subsequent to prostaglandin-induced luteolysis. Prostaglandin analogues were used to induce luteal regression simultaneously in a number of ewes, thereby synchronizing the final stages of follicular maturation in these animals. Some of the ewes were anaesthetized for 24 h immediately after the injection of prostaglandin (experiment 1), and others for 15 h, starting 24 h after the injection of prostaglandin (experiment 2). In both experiments administration of anaesthetic significantly delayed the onset of oestrus and the time of ovulation relative to prostaglandin-treated control animals. The results from assays of blood samples collected at regular intervals in experiment 1 indicated that the preovulatory peak in the concentration of LH and the periovulatory changes in the concentration of FSH were similarly delayed and that during anaesthesia the level of LH was significantly reduced. It is suggested that the reduced level of LH, which probably resulted from a reduction in the secretion of releasing factor due to anaesthesia, failed to support oestrogen production by the Graafian follicle(s), thereby delaying the occurrence of oestrus and ovulation.


Recurrent breeding seasons in pinealectomized or optic-nerve-sectioned ferrets. The breeding seasons of two groups of pinealectomized ferrets and one group of blind ferrets, with appropriate controls, were observed for up to 5 years after the operations. All animals showed recurrent breeding seasons. Both pinealectomized and blind animals became increasingly asynchronous after the first season, but showed no evidence of a free-running, endogenous circannual periodicity nor of synchronization to light or other environmental cues.


Activation of hypothalamic neuronal activity by the electrolytic deposition of iron into the preoptic area. Changes in brain activity after electrochemical stimulation of the preoptic area of pro-oestrous rats were studied by the measurement of the electro-encephalogram (EEG) of the frontal cortex and the recording of single neurones in the anterior hypothalamus. All rats were anaesthetized with urethane between 10.00 and 12.00 h to allow prolonged electrophysiological recording and to block the spontaneous surge of LH during the afternoon. Electrochemical stimulation was applied, between 12.00 and 14.00 h, as an anodal current through an implanted steel electrode; this caused the electrolytic deposition of iron and evoked the release of LH and ovulation. Electrochemical stimulation of the preoptic area changed the cortical EEG, either immediately or after a delay of a few minutes, from a labile pattern with alternate periods of arousal and slow-wave sleep, to a stage of continuous arousal which persisted for the remainder of the recording period (2--3 h). Conversely, the EEG pattern of the cortex was not disturbed by electrolytic lesions placed in the preoptic area through a platinum electrode. Electrochemical stimulation of the arcuate region of the hypothalamus, the lateral septal area, the medial amygdaloid complex and the anterior parts of the thalamus caused no obvious change in the EEG patterns. Ipsilateral anterior hypothalamic neurones, about 1 mm caudal to the focus of electrochemical stimulation, displayed an immediate decrease in electrical activity after application of the current. After 10--20 min however, the rates of discharge of action potentials in 9 out of the 16 neurones under consideration increased progressively from 0.5 to 15--25 action potentials/s and these rates were maintained until the recordings were lost after 90--230 min. No such acceleration in electrical activity was observed in neurones on the contralateral side. Iron deposited during electrochemical stimulation was precipitated as sulphide and stained by Timm's method. There was a central damaged area of radius 0.6 mm surrounded by an 'undamaged' area with considerable infiltration of iron, up to a distance of 1.7 mm from the electrode tip. Cells within the area of infiltration did not stain for iron 10 min after electrochemical stimulation, but after 30 min, neural elements in this peripheral zone were stained in a manner similar to the Golgi method. The concentrations of LH in the plasma remained unchanged in all rats for 10--15 min after electrochemical stimulation. Thereafter, the concentrations increased progressively and approximately in parallel to the changes in action potential activity until, after 2 h, the individual concentrations of 300--600 ng LH/ml were more than six times the values obtained before stimulation. Bilateral electrochemical stimulation resulted in appreciably higher concentrations of LH and produced values close to those observed during the pro-oestrous surge of the hormone...


Detrimental effect of visible light on meiosis of mammalian eggs in vitro. Short wavelength visible light (less than 470-480 nm) emmitted from ordinary light sources is detrimental to unfertilized hamster eggs in that prolonged exposure to the light disturbs the completion of normal meiosis after the eggs are penetrated by spermatozoa. The fluorescent light commonly used in modern laboratories is more harmful than the light from incandescent lamps. In experiments involving the handling of eggs in vitro, minimal exposure to the light or the use of appropriate filters (e.g., red cellophane sheets) is recommended.


Models of infantile undernutrition in rats: effects on milk. Although there are at least six commonly used methods for producing malnutrition in suckling rat pups, no attempt has been made to determine the quality of the nutrients available to the pups in each of these methods. In the present study, milk composition was determined for each of these methods and related to maternal nutritional status. These methods were: 1) large litters were fostered on the dams; 2) the dams were fed low protein diets; 3) the dams were fed a reduced amount of good quality diet; 4) and 5) the pups were removed from the dams and kept 4) with a non-lactating female or 5) in an incubator for half of the day; and 6) some of each dams' teats were destroyed. Maternal nutritional status was affected only in the cases where the dams' diet was deficient (methods 2 and 3), producing weight loss and lowered hematocrits in both cases and lowered serum protein when the low protein diet was fed. The low protein diet condition resulted in a lower nitrogen content and an elevated total fat content of the mild. It was concluded that previous consideration of all of the above methods as equivalent in terms of the quality of the milk produced is not justified.


Relationship of intravenous sodium bicarbonate infusions and cerebral intraventricular hemorrhage. The incidence of cerebral intraventricular hemorrhage was determined by computed tomography in 100 infants with birth weights less than or equal to 1,500 gm. A comparison of IVH with serum sodium concentrations and the amount of intravenous sodium bicarbonate administered did not reveal a significant relationship. Analysis of the method of infusion of sodium bicarbonate indicated that the rapid infusion of hyperosmolar (M to M/12) sodium bicarbonate is associated with a significantly increased incidence of IVH.