Utilization of L-cell nucleoside triphosphates by Chlamydia psittaci for ribonucleic acid synthesis. Long-term, 32-P-labeled L cells were infected with the obligately intracellular parasite Chlamydia psittaci (strain 6 BC). At 20 h postinfection, [3-H]uridine was added, and the infected cells were sampled at intervals for incorporation of the labels into the uridine triphosphate (UTP) and cytidine triphosphate (CTP) pools of the host L cell and the uridine monophosphate (UMP) and cytidine monophosphate (CMP) in 16S ribosomal ribonucleic acid (RNA) of the parasite. The specific activity of the nucleotides was calculated from the ratio of 3-H to 32-P counts in the nucleotides. The rate of approach to equilibrium labeling of UTP and CTP in L-cell pools and UMP and CMP in 16S RNA from the exogenous uridine label was determined from the increase in the ratios of the specific activities of CTP to UTP and CMP to UMP with time. The rate of approach to equilibrium CMP:UMP labeling of the 16S RNA of C. psittaci was consistent with the rate predicted from the kinetics of labeling of the CTP and UTP pools of the host L cell. In analogous experiments, the rate of approach to equilibrium guanosine monophosphate:adenosine monophosphate labeling of 16S RNA from an exogenous [14-C]adenine label was consistent with the rate predicted from the kinetics of labeling of the purine nucleoside triphosphate pool of the host cell. These results support the concept that members of the genus Chlamydia owe their obligate intracellular mode of reproduction to a requirement for energy intermediates which is fulfilled by the host cell. In addition, evidence was obtained that the total acid-soluble purine nucleoside triphosphate pool of L cells accurately represents the precursors of L-cell 18S ribosomal RNA.


Structural similarity of the membrane envelopes of rhizobial bacteroids and the host plasma membrane as revealed by freeze-fracturing. The freeze-fracture technique was used to study the host plasma membrane and the membrane envelope of bacteroids in rhizobial root nodules of three host-rhizobium combinations. In all three combinations studied, the membrane envelopes of bacteroids are structurally similar to their host plasma membrane. However, the membrane appears to be reversed, because the number and arrangement of particles in the outer fractured face (face A, concave) and in the inner fractured face (face B, convex) of the host plasma membrane are seen, respectively, in the inner fractured face (face B, convex) and in the outer fractured face (face A, concave) of the membrane envelope of the bacteroids at an early stage. This reversion of the membrane surface is consistent with the hypothesis that the membrane envelopes of bacteroids are derived from the host plasma membrane during endocytotic engulfment.


Plasmalogen composition of Anaeroplasma. The polar lipids of Anaeroplasma contained 33.1 percent alk-1'-enyl glyceryl ether (plasmalogen) form. Phosphatidylglycerol was the major polar lipid (55.2 percent) and contained nearly all of the plasmalogen. The alk-1'-enyl glyceryl ether form accounted for 58.3 percent of the phosphatidylglycerol.


Crystallization and preliminary x-ray investigation of soybean beta-amylase. Beta-Amylase [1, 4-alpha-D-glucan maltohydrolase, EC 3.2.1.2] has been purified from defatted soybean meal by fractional precipation with ammonium sulfate, ion-exchange chromatography on CM- and DEAE-Sephadex and gel filtration chromatography on Sephadex G-100.  Two different components of beta-amylase were crystallized from ammonium sulfate solutions, and the homogeneity of each preparation was confirmed by sedimentation and disc electrophoretic analyses.  Both components of soybean beta-amylase formed large single crystals (trigonal crystal system) from 40--50 per cent saturated ammonium sulfate solution buffered at pH 5.4 on dialyzing concentrated protein solution in the apparatus of Zeppezauer et al.  Preliminary X-ray diffraction data gave a hexagonal lattice with unit cell dimensions a=86.1 A and c=144.4 A.  The space group corresponds to P3121 or P3221, and one asymmetric unit contains one molecule of beta-amylase, assuming a crystal density of 1.25 g/ml and a molecular weight of the enzyme of 60,000 daltons.  In this case, the crystal has a volume of 2.53 A-3 per atomic mass unit, and the percentage of protein in the crystal is about 52.


Utilization of ribonucleic acid and deoxyoligomer primers for polyadenylic acid synthesis by adenosine triphosphate: polynucleotidylexotransferase from maize. The ATP:polynucleotidylexotransferase isolated and purified from maize seedlings catalyzes the synthesis of polyadenylic acid by the sequential addition of 80 to 200 AMP moieties from ATP to the 3'-hydroxyl terminus of either ribo- or deoxyoligomers. Copurification of the RNA and DNA-primed activities, identical metal cofactor and reaction requirements for either primer and identical heat inactivation curves with either primer strongly suggest that both primers are utilized by the same enzyme.


The N, O-diacetylmuramidase of chalaropsis species. IV. Tryptic peptides. Lysozyme Ch was hydrolyzed with trypsin in 2 M urea and the resulting peptides were separated by a combination of gel filtration and ion exchange chromatography. Ten peptides and free lysine were produced by tryptic action. The enzyme has 5 arginine and 4 lysine residues per molecule and one of the peptides arose from a chymotryptic-like cleavage of a tyrosyl-seryl bond near the amino-terminal end of the enzyme. The entire molecule is accounted for by the tryptic peptides, which have been ordered withing the peptides obtained by cyanogen bromide cleavage of the molecule.


The N, O-diacetylmuramidase of Chalaropsis species. V. The complete amino acid sequence. The complete amino acid sequence of lysozyme Ch has been established by a combination of automated and manual Edman degradation and carboxypeptidase digestion.(see article)There is a single disulfide bond in the center of the molecule. The enzyme has 211 residues with a calculated molecular weight of 22,415. Lysozyme Ch has an amino acid sequence that is totally different from all other lysozymes whose sequences are known.


Mechanism by which antibodies inhibit hapten-malate dehydrogenase conjugates. An enzyme immunoassay for morphine. Antimorphine antibodies inhibit the activity of morphine conjugates of mitochondrial malate dehydrogenase. Conjugation of malate dehydrogenase through tyrosine and amino groups resulted in only moderate losses of enzyme activity. On conjugation through disulfide bonds the enzyme activity first increased but dropped sharply with increasing substitution. Only the former conjugates were inhibited by excess antibodies. The degree of inhibition (up to 86%) was directly related to the number of morphine residues bonded directly to amino groups. The maximum number of antibody binding sites that bind to enzyme was nearly equal to the number of haptens provided there were 16 or less haptens/enzyme. However up to 26 haptens/enzyme became completely bound by antibody on long incubation. Inhibition of enzyme activity was detectably reduced by 2 times 10 minus 9 M morphine or 2 times 10 minus 10 M codeine, thus providing a sensitive assay for these drugs. The data suggest that enzyme inhibition occurs by conformational freezing of the enzyme when antibody binds to a morphine residue attached to one specific amino group.


Reversible inactivation of pancreatic deoxyribonuclease A by sodium dodecyl sulfate. Removal of COOH-terminal residues from the denatured protein by carboxypeptidase A. In the course of experiments on the role of the COOH-terminal residues in pancreatic deoxyribonuclease, we undertook to ascertain whether the presence of sodium dodecyl sulfate would render the normally unavailable terminus susceptible to hydrolysis by carboxypeptidase A. When DNase A is dissolved in 0.005% sodium dodecyl sulfate the protein becomes enzymically inactive when assayed against DNA in the same sodium dodecyl sulfate concentration. The loss of activity caused by treatment with sodium dodecyl sulfate for 1 hour at 45 degrees can be fully restored if the detergent-containing solution is diluted 10-fold into 6 M guanidinium chloride and then 10-fold into a pH 7.0 buffer, 10 mM in CaCl2, prior to a 100-fold dilution for assay. The presence of Ca2+ is essential for the refolding process. If the same degree of dilution is made into sodium dodecyl sulfate-free buffer without the guanidinium chloride step, there is very little reversal of the inactivation. An almost complete loss of regenerable activity is caused by 1 hour of digestion by carboxypeptidase at 45 degrees in the presence of 0.03% sodium dodecyl sulfate. Although up to 6 amino acid residues can be removed from the COOH terminus, the loss of activity can be correlated with the removal of either 1 or 2 amino acid residues (-Leu-Thr) from the COOH-terminal sequence. Thus, DNase A is one of the several enzymes in which residues at the COOH terminus are essential to the active conformation. If the enzyme minus 2 to 6 terminal residues was mixed with a 15-residue COOH-terminal peptide (obtained by cyanogen bromide cleavage), only about 2% activity could be regenerated.


Sequential phsophorylation of histone subfractions in the Chinese hamster cell cycle. Ion exchange chromatography and preparative electrophoresis were used to examine the phosphorylation of histone f1 and f3 subfractions in synchronized Chinese hamster cells (line CHO). Three discrete f1 phosphorylation events were demonstrated to occur in sequence during the cell cycle. The first event (f1G1) commenced in G1 2 hours prior to entry of cells into S phase; the second event (f1s) commenced simultaneously with initiation of DNA synthesis; and the third event (f1M) commenced when cells entered mitosis. F1M phosphorylation occurred simultaneously with the phosphorylation of histone f3 (which is not phosphorylated during G1, S, or G2). Fractionation of f1 and f3 revealed no differences in these sequential phosphorylation patterns among the various f1 and f3 subfractions, indicating that these phosphorylations are general biochemical events of the cell cycle. Phosphorylated (f1G1) was found to accumulate in cells as they traversed THEIR CELL CYCLE. F1s was phosphorylated to twice the extent of f1G1, but f1s did not accumulate in the cells as they passed through interphase. F1M was phosphorylated to about 4 times the extent of the first phosphorylated form (f1G1). A model of the relationship of histone phosphorylation to the cell cycle is presented which suggests that (a) f1G1 phosphorylation is involved with chromatin structural changes necessary for cell proliferation; (b) f1s phosphorylation is involved with DNA replication; (c) F1M and f3 phosphorylations are involved in chromosome condensation.


Biosynthesis of chondroitin sulfate proteoglycan. Subcellular distribution of glycosyltransferases in embryonic chick brain. The subcellular distributions of five glycoslytransferases involved in the biosynthesis of the chondroitin sulfate proteoglycan and of a sixth glycosyltransferase, presumably involved in glycoprotein biosynthesis, were examined in 13-day chick enbryo brain. Fractionation studies performed by the procedure of Gray and Whittaker (Gray, E.G., and Whittaker, V.P. (1962) J. Anat. (London) 96, 79-88) revealed that three of the six enzymes were directly associated with the membrane fraction of synaptosome-enriched preparations; varying amounts of the remaining glycosyltransferases were distributed between the 100,000 times g supernatant and the synaptosome-enriched fraction after differential and density gradient centrifugation of crude chick brain homogenates. The time of appearance of three of the glycosyltransferases was examined in chick embryo brain tissue at several stages of development. The brain content of each glycosyltransferase increased rapidly between day 7 and hatching at day 21. A sharp decline in each of the glycosyltransferase activities occurred at hatching.


Activation of the action potential Na+ ionophore of cultured neuroblastoma cells by veratridine and batrachotoxin. The activation of the action potential Na+ ionophore by veratridine and batrachotoxin is time- and concentration-dependent and completely reversible. Batrachotoxin acts more slowly than veratridine. The concentration dependence of activation at equilibrium suggests reversible interaction of each toxin with a single class of independent sites having dissociation constants at physiologic ion concentrations of 80 plus or minus 13 muM for veratridine and 0.4 plus or minus muM for batrachotoxin. The maximum velocity of Na+ uptake at 50 mM Na+ is 128 plus or minus 12 nmol/min/mg in the presence of batrachotoxin compared to 48 plus or minus 4 nmol/min/mg in the presence of veratridine. Treatment of cells with excess veratridine in addition to batrachotoxin inhibits batrachotoxin-dependent 22-Na+ uptake. The concentration dependence of this inhibition suggests that it reflects competitive displacement of batrachotoxin from its binding site by veratridine. The activation by veratridine and batrachotoxin is inhibited in a competitive manner by divalent cations. The inhibition by divalent cations exhibits significant ion specificity with Mn-2+ greater than Co-2+ greater than Ni-2+ greater than Ca-2+ greater than Mg-2+ greater than Sr-2+. The inhibition constants (KI) for Ca-2+ are 0.84 mM for veratridine-dependent 22-Na+ uptake and 1.2 mM for batrachotoxin-dependent 22-Na+ uptake. The activation by veratridine and batrachotoxin is inhibited in a noncompetitive manner by tetrodotoxin. The apparent KD for tetrodotoxin as 11 plus or minus 1 nM in the presence of 150 mM Na+ and approximately 8.5 nM in 50 mM Na+. Divalent cations do not affect the apparent KD for tetrodotoxin. A hypothesis is presented which suggests that batrachotoxin, veratridine, and divalent cations interact with an activation site associated with the action potential Na+ ionophore, whereas tetrodotoxin interacts with a physically and functionally independent site involved in the transport of monovalent cations by the ionophore.


Location of procollagen in chick corneal and tendon fibroblasts with ferritin-conjugated antibodies. Three distinct antiprocollagen preparations were characterized and used in immunocytochemical staining of chick embryo corneal and tendon cells. The several ferritin-conjugated antibody preparations permitted similar location of procollagen in the cisternae of the rough endoplasmic reticulum and in Golgi elements in both cell types. The ability to demonstrate and interpret specific ferritin staining was dependent on the extent of membrane breakage in each of those organelles, coupled with adequate retention of cell morphology. Corneal fibroblasts appeared to suffer more extensive intracellular membrane damage under controlled conditions of homogenization than tendon fibroblasts, facilitating the identification of procollagen in Golgi vacuoles of these cells. None of the labeled material appeared to by cytoplasmic in origin since ferritin was observed in the cytoplasm only in the vicinity of Golgi elements that were extensively broken. This study extends previous immunological evidence for the presence of procollagen in the Golgi complex and calls attention to the problems to be encountered in locating the antigen in small Golgi vesicles and lamellae.


Studies of murine erythroid cell development. Synthesis of heme and hemoglobin. Techniques of cell separation were used to isolate murine erythroid precursors at different states of maturation. Cells were studied before and after short-term incubation in the presence or absence of erythropoietin. Complementary results were obtained by direct examination of the cell fractions and by the short-term culture experiments. Indices of heme synthesis, including incorporation of 59Fe or [2-14C]glycine into heme and activity of delta-aminolevulinic acid synthetase, were already well developed in the least mature cells, chiefly pronormoblasts. Activity then rose moderately in the cell fractions consisting primarily of basophilic and polychromatophilic normoblasts, and fell off with further increases in cell maturity. On short-term culture in the presence of erythropoietin, activity declined with increasing cell maturation except in the least mature fraction where the original level of activity was maintained. By contrast, synthesis of labeled hemoglobin ([3H]leucine) was very low in the least mature cell fractions and rose progressively with increasing cell maturity. The rate of hemoglobin synthesis increase in cells at all stages of maturation when cultured in the presence of erythropoietin. Despite the different patterns observed for heme synthesis and hemoglobin synthesis, both synthetic activities were consistently higher in cells cultured with erythropoietin as compared to controls. These findings suggest that erythropoietin stimulates biochemical differentiation of erythroid precursors at various stages of maturation. They also demonstrate an asynchronism between heme synthesis and hemoglobin syhthesis; heme synthesis is already well developed in the least mature erythroid cells and begins to diminish as the capacity for hemoglobin synthesis continues to rise.


Studies on microperoxisomes. VII. Pigment epithelial cells and other cell types in the retina of rodents. The pigment epithelial cell of the retina actively participates in two aspects of lipid metabolism: (a) the fatty acid esterification of vitamin A and its storage and transport to the photoreceptors, and (b) the phagocytosis and degradation of the lipoprotein membrane disks shed from the photoreceptor cells. Study of the pigment epithelial cells of adult albino and pigmented rodents has revealed the abundance of an organelle, microperoxisomes, not previously known to exist in this cell type. The metabolism, transport, and storage of lipids are major functions of other cell types which possess large numbers of microperoxisomes associated with a highly developed smooth endoplasmic reticulum. Microperoxisomes were encountered, but relatively rarely, in Müller cells and vascular endothelial cells. A tubular system in photoreceptor terminals is reactive in the cytochemical procedure used to visualize microperoxisomes.


Transport and storage of serotonin by thrombin-treated platelets. Repeated thrombin treatment of washed platelets prepared from rabbits can decrease the serotonin content of the platelets by about 80%. When these platelets are deaggregated they reaccumulate serotonin but their storage capacity for serotonin is reduced by about 60%. If thrombin-pretreated platelets are allowed to equilibrate with a high concentration of serotonin (123 mu M), they release a smaller percentage of their total serotonin upon further thrombin treatment, in comparison with the percentage of serotonin released from control platelets equilibrated with the same concentration of serotonin calculations indicate that in thrombin-treated platelets reequilibrated with serotonin, two-thirds of the serotonin is in the granule compartment and one-third is in the extragranular compartment, presumably the cytoplasm. Analysis of the exchange of serotonin between the suspending fluid and the platelets showed that thrombin treatment does not alter the transport rate of serotonin across the platelet membrane and does not cause increased diffusion of serotonin from the platelets into the suspending fluid. The primary reason for the reduced serotonin accumulation by the thrombin-treated platelets appears to be loss of amine storage granules or of the storage capacity within the granules.


Nucleolar necklaces in chick embryo fibroblast cells. I. Formation of necklaces by dichlororibobenzimidazole and other adenosine analogues that decrease RNA synthesis and degrade preribosomes. A number of chemicals, mostly adenosine analogues, cause the nucleolus of the chick embryo fibroblast to lose material and unravel over a period of several hours into beaded strands termed nucleolar necklaces (NN). The results of analyses of the fibroblasts, treated with the NN-forming chemical dichlororibobenzimidazole (DRB), suggests that the following biochemical alterations occur: DRB almost completely prevents the increase in both messenger RNA (mRNA) and heterogeneous nuclear RNA. It interferes with ribosome synthesis by decreasing the rate of 45S ribosomal RNA (rRNA) accumulation by 50%, slowing the rate of 18S rRNA appearance by 50%, and causing an extensive degradation (80%) of the 32S and 28S rRNA-containing preribisomes. Most of this preribosome degration probably occurs at or before the 32S rRNA preribosome stage. The degradation of these preribosomes appears to be due to the formation of defective 45S rRNA preribosomes rather than to a direct DRB interference with preribosome processing enzyme action. DRB inhibits total cellular RNA synthesis in less than 15 min, suggesting a direct interference with RNA synthesis. DRB also inhibits the uptake of nucleosides into the cell. DRB in the concentrations used does not appear to directly interfere with the translation of mRNA (i.e., protein synthesis). Other NN-forming adenoside analogues and high concentrations of adenosine (2 mM) cause biochemical alterations similar to those produced by DRB. To explain the preribosome degradation, we propose the hypothesis that DRB inhibits the synthesis of mRNA; as a consequence, some of the preribosomal proteins that normally coat the 32S rRNA portion of the 45S precursor RNA become limiting, and this defective portion is then subject to degradation by nucleases.


Selection of mammalian cells resistant to a chloramphenicol analog. This study describes the selection and preliminary characterization of mammalian cells resistant to 100 mug Tevenel/ml. Tevenel, the sulfamoyl analog of chloramphenicol, is a specific inhibitor of mitochondrial protein synthesis. After growth in suspension culture for 5 days in 100 mug Tevenel/ml and subsequent plating in 100 mug Tevenel/ml, LMTK- cells yielded resistant clones. As a control, L cells treated identically yielded no clones. Three resistant clones were chosen for study. Each resistant cell line had an identical growth rate in the presence and absence of 100 mug Tevenel/ml. By plating efficiency analysis, the resistant cells were found to be cross-resistant to D-chloramphenicol. The change responsible for resistance was found to be stable for at least 100 generations in the absence of the drug. Protein synthesis by isolated mitochondria of resistant cells was found to be less inhibited by concentrations of both Tevenel and D-chloramphenicol up to 200 mug/ml than the protein synthesis by LMTK- mitochondria. This resistance in vitro was not changed by incubation of the mitochondria in 0.01% Triton X-100.


Synthesis of ribosomal RNA in synkaryons and heterokaryons formed between human and rodent cells. A study has been made of the ribosomal RNA and chromosome constitution of man-mouse hybrid cells. Previous work has shown that no human 28s rRNA is detectable in man-mouse synkaryons. In general human chromosomes are lost from such hybrids. With a recently developed method for distinguishing mouse from human chromosomes, an analysis of various man-mouse hybrid cell lines has been made. This indicates that not all the human chromosomes bearing nucleolar organizers are lost in the hybrid cells and such loss cannot alone explain the absence of human 28s rRNA. An examination of the 28s rRNA synthesized by heterokaryons formed from several different parent cells has revealed that both parental types of 28s rRNA are present in heterokaryons. The control of rRNA synthesis in hybrid cells is discussed.


Effects of cytochalasin B on the aggregation, electrophoretic mobility and surface morphology of chick neural retina cells. Over a concentration range of o-5-10 mug/cm-3, cytochalasin B caused a biphasic change in the electrophoretic mobility of disaggregated neural retina cells. An initial rise in anodal mobility at low concentrations of the drug was transformed into a reduction in the mobility below that of the control at a concentration of 10 mug/cm-3. The effect of cytochalasin B was found to be reversible by washing treated cells in cytochalasin B-free media. This was investigated at a concentration of cytochalasin at which the greatest difference existed between the mobilities of the control and experimental cell suspensions. Reaggregation of cell dispersions failed to show any significant difference in the rate of aggregation between cytochalasin B-treated cells and the control. Scanning electron microscopy of cells fixed while in suspension also showed little significant change in the surface morphology upon application of cytochalasin B. In high concentrations of the drug cells appeared somewhat smoother in outline, but no correlation was found between changes in surface morphology and the variations in cell electrophoretic mobility. It is concluded that the observed changes in electrophoretic mobility may be attributed to a binding of cytochalasin B to the cell membrane. This lends support to the hypothesis that the primary site of action of cytochalasin B may be the plasma membrane.


The quantitative requirements of human diploid cells (strain MRC-5) for amino acids, vitamins and serum. The uptakes of all essential amino acids, vitamins (except riboflavin), glucose and serum during growth of human diploid cells (MRC-5) were determined. The amino acid uptakes varied considerably with the conditions of culture. The glucose requirement is several times greater than that for mouse LS or human HeLa cells. These analytical results were used to modify the medium so as to ensure that an excess of all defined medium constituents was present and pH was not limiting during study of the serum requirements. It was then found that maximum cell populations were directly proportional to the serum concentration. Hence the growth was limited by the supply of an unknown growth factor in serum. The serum growth factor was not replaced by a mixture of over 60 vitamins, co-enzymes, hormones and other organic and inorganic compounds considered to be possible growth factors, although this mixture did not lower the growth rate and somewhat (22%) increased the yield from the serum growth factor. The unit of serum growth factor is precisely defined in terms of the amount in a standard batch of calf serum. This standard contains 10 units/ml whereas the other batch of serum used contained only 5 units/ml.


Twenty-four-hour prolactin profiles in normal and disease states: failure of thyroxine to modify prolactin secretion. In order to assess the role of thyroid hormone on physiologically and pharmacologically induced prolactin (PRL) secretion, serum PRL concentrations were measured in 4 normal women and 4 women with various endocrinopathies before, and 4 to 6 days following, the ingestion of L-thyroxine (T4). A single 1.5 to 3.0 mg dose of oral T4 produced approximately a 2-fold increase in serum T4. Exogenous T4 did not significantly alter the mean concentration, or the pattern of PRL secretion during a 24-h interval in either normal individuals or 3 patients with galactorrhea. The lactating patients had elevated basal PRL levels and a blunted secretory response to intramuscular chlorpromazine; however, neither fasting PRL nor the peak response to chlorpromazine was altered by T4. L-Dopa suppression of serum PRL was not significantly influenced by T4 in these patients. In conclusion, PRL secretion remained unaltered after the administration of thyroxine in doses sufficient to produce approximately a 2-fold increase in serum T4. This challenges the concept that T4 and TRH are important physiologic regulators of PRL secretion.


Influence of timing and intensity of musclar exercise on temporal patterns of plasma cortisol levels. Plasma concentrations of cortisol were measured in 9 subjects form 0800 h to 1800 h at 10-min intervals during resting periods and 5 min intervals during exercise and recovery. This was done to assess to effect of exercise on the patterns of plasma cortisol levels. At 1000 h, 90 min exercise at a moderate work level (55% of Vo2 max) produced a transient rise in plasma cortisol which averaged 11.9 pg/100 ml (SE plus or minus 1.2) and introduced subsequent suppression of meal-related increase, At a lower work load (25% of Vo2 max) a similar transient rise occurred, avering 11.7 pg/100 ml  (SE plus or minus 2.2); latency rate of concentration change and magnitude and not significatntly different from those at a moderate work load. A strenous work level was required to produce a more rapid response with an increased secretion rate. When 90 min exercise at a moderate work levels was preformed at 1300 h. (i.e. coinciding with postprandial cortisol concentration peak), the increase in plasma cortisol concentration (1.3 pg/100ml (SE plus or minus 0.3) was significantly lower than that induced by the same exercise performed in the morning. These results demonstrate that high plasma levels of cortisol diminish the subsequent stress response and that exercise-induced and meal-induced increases are not active.


Long-term effects of vasectomy on pituitary-gonadal function in man. Plasma concentrations of testosterone (T), follicle stimulating hormone (FSH), and luteinizing hormone (LH) and percent binding of T to plasma protein (TB) were studied in 16 normal fertile men and in 81 men who had undergone vasectomy from 1 to 5 yr earlier. No significant difference from the control values to T = 582 plus or minus 39.5 (SE) ng/100 ml; FSH = 15.09 plus or minus 2.18 (SE) mIU/ml; LH =10.03 plus or minus 1.13 (SE) mIU/ml and TB = 92.99 plus or minus 0.285 (SE) % binding was detected in any group of the vasectomized men in any of the parameters studied.


A simple specific assay for estriol in maternal urine. In medical centers with many high risk pregnancies, the ability to perform a large number of urinary estriol measurements is required. The analytical procedure should provide simplicity, specificity, and economy. A solid phase radioimmunoassay is presented which utilizes antibody generated against an estriol 16-glucuronide-bovine serum albumin conjugate. The assay allows analysis of the principal component of maternal urinary estriol without hydrolysis or purification, with adequate specificity and the capacity for processing large numbers of samples.


Isolation, assay, and secretion of individual human neurophysins. Human neurophysin was isolated from acetone-dried human posterior pituitaries and separated into two major neurophysin peptides by ion exchange chromatography and into four major peptides by preparative disk gel electrophoresis. Antisera raised in rabbits distinguished only two specific antigenic sites on the isolated neurophysin peptides. Individual sensitive and specific radioimmunoassays for two human neurophysins were developed. These assays were used to measure each neurophysin in unextracted human plasma. The two neurophysins are secreted independently in man. One assay measures a neurphysin that is specifically secreted in response to estrogen administration, estrogen-stimulated neurophysin (ESN). The other assay measures a neurophysin that is specifically secreted in response to cigarette smoking, nicotine-stimulated neurophysin (NSN). The mean ESN is 1.1 ng/ml plus or minus 0.7 SD in women and 1.0 ng/ml plus or minus 0.7 SD in men. The mean NSN is 0.9 ng/ml plus or minus 0.2 SD in women and 0.6 ng/ml plus or minus 0.3 SD in men. It is proposed that these may prove to be a specific human "oxytocinneurophysin," ESN, and a human "vasopressin-neurophysin," NSN.


Postprandial blood glucose and insulin in cows fed high grain. The effects of ration on postprandial serum glucose and insulin were detemined in 12 lactating Holstein cows. Six were fed a high grain ration of 15% hay and 85% concentrate (dry basis) and the other six a control ration of 55% corn silage, 10% hay and 35% concentrate. High grain feeding increased glucose and insulin at all hours postfeeding as compared to control cows. In the cows fed high grain, glucose increased from 63.3 to 72.2 mg/100 ml and insulin from 19.2 to 25.6 mu units/ml serum just before feeding to 3 h postfeeding. Values for 2, 3, and 4 h samples were greater than for other sampling time. Serum glucose was 55.5 mg/100 ml at 1 h in control cows which was above other sample. Serum insulin followed a pattern similar to glucose in controls but was not signigicantly different with time. The blood changes due to high grain feeding are probably related to low milk fat production.


Release of oxytocin in the cow during milking. Jugular vein oxytocin concentrations were determined at 15-s intervals on 10 Holstein cows during milking. Oxytocin ranged from .5 muU/ml plasma prior to udder washing to 652.7 muU/ml plasma at 1 min after application of teat cups. The results indicate a gradually increasing release of oxyctocin after stimulation and a maximal oxytocin release 1.5 min after start of udder washing and 1 min after teat cup application.


Immunity to Trichinella spiralis VI. The specificity of the immune response stimulated by the intestinal stage. Mice were immunized to the intestinal stage of T. spiralis by using infections terminated with methyridine before production of newborn larvae had commenced. The muscle larvae which encysted following a normal complete challenge infection were reduced by 87 and 95% in immunized mice. No statistically significant reduction in a challenge infection of intravenously injected parenteral larvae was produced (8% and 15% actual reduction). Previous work has shown that adult worms in a challenge infection are stunted and expelled earlier as well as having a reduced fecundity; it is concluded that the immunity generated by the intestinal stage is largely specific in its action to that phase in a challenge infection.


Polysaccharides as labels for antibodies in electron microscopy. Two polysaccharides, dextran 250 and dextran 70, were covalently linked to antibody molecules, antihuman immunoglobulin G and antihuman type O red blood cells. In electron microscope preparations exposed to lead citrate, polysaccharides, because they chelate lead, were quite dense. Polysaccharides served as a tag for the antibody molecules. Also, bacterial dextran 1355 was used to demonstrate antibody molecules on the surface of ascites tumor cells which are known to be producing a specific antibody to bacterial dextran 1355. The varying sized polysaccharide molecules that are readily available commercially, the high electron density of the polysaccharides after lead staining and a mild procedure for covalently linking polysaccharide to antibody make polysaccharides attractive as particulate labels for antibody in electronmicroscopy.


Agarose beads as matrices for proteins in cytophotometric investigations of immunohistoperoxidase procedures;. Quantitative aspects of direct immunohistoperoxidase procedures were studied in a model system consisting of agarose beads to which antigens or antibodies had been coupled. It could be proven that the final amount of reaction product resulting from the histoperoxidase reaction with 3,3-diaminobenzidine-tetra HCl in a bead was linearly related to the volume of the beads and to the staining time. This implies that protein-coupled agarose beads are a suitable model for the study of stoichiometric aspects of immunologic reactions in immunohistochemistry as well as in general immunologic methods when peroxidase is used as the protein marker.


Dehydrogenase activity of liver parenchyma in mice exposed to arsenic in drinking water. The effect of arsenic administered in drinking water in the form of sodium arsenite in concentrations of 5,50 and 250 mg As-3+/1 on dehydrogenase activity (DHA) of liver parenchyma homogenate of exposed mice was studied. After an exposure lasting 2, 4, 8, 16, 32, and 64 days, 8--10 exposed mice and 5--10 animals of the control series were studied at each interval. DHA was determined by means of tetrazolium chloride reduction to formazan whose quantity was assessed spectrophotometrically after extraction with butanol. At a concentration of 5 mg A3+/1 a moderate increase in DHA was observed which was replaced by a moderate decrease at the following intervals. At a concentration of 250 mg A3+/1 a deep decrease in DHA (p smaller than 0.05) was recorded which did not return to values compararable with those of the control series in spite of its fluctuating increase in the course of exposure. At a concentration of 50 mg A3+/1 in drinking water ensuring the supply of doses which induce an increase in the tolerance to arsenic a decrease in DHA was found only at the first exposure interval. DHA is thus substantially less altered in comparison with the metabolic consumption of oxygen (MCO) under the same conditions. This finding signifies that MCO is a more sensitive indicator of arsenic exposure than DHA in the used arrangement of the experiment.


[Epidemiology of Dientamoeba fragilis Jepps and Dobell, 1918. 3. Further studies on Enterobius transmission through eggs]. In two new attempts to infect human subjects with pin-worms that were infested, at that time, with protozoa Dientamoeba fragilis, presence of these protoza in the intestinal canal was observed in one case for more than 84 days, and in another case for more than 31 days after contamination. The duration of latent periods fixed for Enterobius vermicularis was almost identical (51 and 50 days). New attempts implement and confirm the results of the first successful attempts to transfer the infection (Ockert, 1972b) as well as the hypothesis of the epidemiological interrelation between both parasites (Burrows and Swerdlow 1956).


Mass occurrence of Yersinia enterocolitica in two establishments of collective care of children. Mass occurrence of intestinal disorders caused by Yersinia enterocolitica simultaneously in two groups of children of under-school age is reported. The question of the source of infection and the routes of transfer are discussed and the various forms pointed out which Y. e. infection may take in under-school children within the framework of an epidemic occurrence in a group of children. The fact is emphasized that the results of serological examination using the method of indirect haemagglutination may be an important guide for the orientation in the epidemiological situation and, in some cases, also in making etiological diagnosis when the proof of Y. e. by culture presents difficulties.


Protection of drinking water sources against modern pollutants. One of the main tasks of hygiene necessitating urgent solution is the regulation, within the framework of governmental planning, of sanitary protection against modern pollutants of sources and water works supplying the human collectivities. Pollution by pesticides, surfactants, aromatic polycyclic hydrocarbons, oil waste products, phenolics, radioactive isotopes etc., is requiring particular concern. Cases of penetration of these chemical substances, especially of pesticides, oil residues and other kinds of industrial waste into ground waters have been recently observed with increasing frequency. Pollution of underground waters occurred either through the storage on the surface ground or into soil of waste products containing these substances or through their discharge into the water stream. The presence of these substances in most water supply systems the lowered effectivity of water-works in the removal of these substances, continual increase in the number of industries producing these pollutants, their participation in the origin or increased incidence of some chronic diseases characterizing the development of modern society make necessary scientific substantiation and urgent solution of the problem.


Ultrastructure of rabbit lung intravenously infected with Mycobacterium simiae "Weiszfeiler" no 52. M. simiae "Weiszfeiler" strain No. 52 belongs to the second Runyon Group of Atypical Mycobacteria characterized by rough, eugonic, yellow-orange pellicle grown on Sauton medium incubated for 14 days at 37 degrees C. A suspension containing 10 mg/ml (equalling 5 times 10-7 viable units) injected intraveneously into rabbits provokes profound changes in the ultrastructure of the pulmonary tissue, manifested in activation of macrophages and pathologic alteration of pneumocytes. In histologic sections stained according to the Ziehl-Neelsen technique and examined in a light-microscope cytoplasmic acid-fast, highly dispersed granules are found in macrophages, resembling in morphological appearance the cellular phospholipids in sarcoid granuloma. Abundant production of interalveolar and interseptal fibrotic collagen tissue was observed in rabbits infected with M. Simiae a rather striking phenomenon in fresh experimental mycobacterial infection in the rabbit. The strain in question should therefore be considered not as Saprophyte but as a Mycobacterium Species with attenuated virulence, closely related to Prissic and Masson's M. scrophulaceum.


The mechanism of damage to neutrophils in allergic reactions in tuberculosis. Allergic vulnerability of neutrophils due to pulmonary antigen and tuberculin in tuberculosis is connected with the fact that the immunological conflict antigen-antibody takes place on the surface of leukocytes. These two effects have a similar mechanism. The tissue antigen or tuberculin adsorbed on the cell surface with the participation of the complement may react with the corresponding circulating antibodies, thus producing damage to neutrophils.


Human lymphocyte cytotoxicity against mumps virus-infected target cells. Requirement for non-T cells. Subpopulations of human lymphocytes were tested for their capacity to kill mumps virus-infected target cells in a 51-chromium release asaay. Using two different cell fractionation techniques, lymphocytes were fractionated into T cell-enriched (primarily T cells) and T cell-depleted (primarily B cells) subpopulations. Filtration of lymphocytes through columns coated with human immunoglobulin and rabbit anti-human-immunoglobulin (Ig-anti-Ig) rendered the resulting T-cell preparation inactive as effector cells against target cells carrying mumps virus antigens. In the second technique, lymphocytes were fractionated by centrifugation into two fractions according to their ability to form spontaneous rosettes with sheep erythrocytes (E). The E-rosette-forming population (primarily T cells) was shown to lack cytotoxic activity against mumps virus-infected target cells. This activity was present in the nonrosetting population. The results suggest that the effector cells involved in this cytotoxic system are of a non-T variety.


Evaluation of human oral organisms and pathogenic Streptococcus for production of IgA protease. IgA protease is a proteolytic enzyme found in whole human saliva and in dental plaque that cleaves both secretory and myeloma IgA of human origin to yield intact Fabalpha and Fcalpha fragments. To determine which bacteria are capable of producing this enzyme, we have examined a variety of strains normally found in the human oral cavity and a number of streptococci of known Lancefield group serotype. Streptococci of groups A, B, C, D, F, G, H, M, and N, Streptococcus mutans, Streptococcus sanguis, Streptococcus mitior, Streptococcus salivarius, Streptococcus faecalis, Veillonella, Lactobacillus, Actinomyces, Propionibacterium, Bacteroides, and Fusobacterium were grown in liquid medium, and fluids were examined for IgA protease activity. Only S. sanguis and clinically isolated group H streptococci elaborated IgA protease under the culture conditions used. Negative strains could not be stimulated to produce the enzyme when cultured in the presence of secretory IgA. Among the natural oral bacteria, capacity to produce IgA protease is restricted to certain species of Streptococcus, notably those of the group H serotype. Since secretory immunity is mediated by the IgA class of antibody, the presence of this enzyme at mucosal surfaces could modify the secretory immune function.


Characterization of two steroidal ketones and two isoprenoid alcohols in dairy products. Two steroidal ketones, delta-4-cholesten-3-one and delta-3,-5-cholestadiene-7-one, were isolated and identified for the first time in anhydrous milk fat and in nonfat dry milk. Together with these, two isoprenoid alcohols, phytol and dihydrophytol, were identified in anhydrous milk fat. Their identities were established on the basis of chromatographic and mass spectral data and confirmed by comparison with authentic materials.


Distribution of molecular species of sphingomyelins in different parts of bovine digestive tract. Sphingomyelins were isolated from mucosal layers of bovine rennet stomach, duodenum, jejunoileum, and colon ascendens. The ceramides obtained after phospholipase degradation were characterized by thin-layer chromatography, mass spectrometry, and gas-liquid chromatography. The main ceramide group from all regions consisted of dihydroxy long-chain bases and normal fatty acids. Sphingosine was the predominant base in all these fractions, and only in rennet stomach were smaller amounts of the C17 and C20 homologs present. Normal saturated C16, C18, C22, and C24 fatty acids were most abundant. In rennet stomach there was in addition a ceramide group having dihydroxy long-chain bases in combination with hydroxy fatty acids. Sphingosine was the predominant long-chain base and the fatty acids were 2-hydroxy C16, C22, C23, and C24. From jejunoileum three minor ceramide fractions were isolated; these consisted of phytosphingosine and normal fatty acids C22-C24), sphingosine and 2-hydroxy fatty acids (C16-C24), and phytosphingosine and 2-hydroxy fatty acids (C22-C24), respectively. No branched paraffin chains were found in significant amounts. Sphingomyelins with trihydroxy long-chain bases and 2-hydroxy fatty acids found in jejunoileum were also detected in bovine kidney and have not been demonstrated before. These sphingomyelins from both kidney and jejunoileum showed a preferential combination of trihydroxy bases and fatty acids with very long chains (C22-C24).


Humoral immunostimulation. IV. Role of complement. When L cells were treated with anti-L-cell antibody in medium containing heat-inactivated fetal calf serum, nucleoside uptake and cell growth were stimulated. The response was markedly increased when fresh, unheated sera from calves, guinea pigs, humans, mice, or rabbits were also present. The factors in unheated serum responsible for the enhancement of immunostimulation were studied. Using low concentrations of sera deficient in various complement (C) components and low concentrations of antibody no augmentation of immunostimulation was seen with Clr-deficient human serum, C2-deficient human serum, C2,4-deficient human serum, C4-deficient guinea pig serum, C3-C9-depleted guinea pig serum (by administration of cobra venom factor to animals), but stimulation was observed with C5-deficient human serum, C5-deficient mouse serum, and C6-deficient rabbit serum. When the concentration of anti-serum was raised, however, augmentation was observed with C4-deficient guinea pig serum. Thus, at low concentrations of antiserum enhancement appeared to occur through the classical C pathway, whereas at high concentrations of antibody either the classical or alternate C pathways appeared to be involved. Stimulation was specifically restored by purified C2 in C2-deficient serum and by C3 in C3-C9-deficient serum. Under the usual reaction conditions consumption of guinea pig C component C4 could be demonstrated which provided direct evidence for activation of the classical C pathway under conditions leading to immunostimulation. By immunofluorescence, cells treated with antibody and normal human serum had human C3 deposited at the cell surface. Taken together these observations suggest that C activated through C3 by either the classical or alternate pathways has the potential to enhance nucleoside incorporation into DNA and cell growth of cells exposed to limiting amounts of antibody. Although the mechanism of stimulation is unknown, it is likely to involve a direct effect of C3 at the level of the cell membrane.


Subcellular platelet factor VIII antigen and von Willebrand factor. Subcellular membrane and granule fractions derived from human platelets contain factor VIIII antigen and von Willebrand factor activity but not factor VII procoagulant activity. Circulating platelets constitute a significant reservoir of plasma factor VIII antigen, containing approximately 15% of the amount of factor VIII antigen present in platelet-poor plasma. The antibiotic ristocetin, which aggregates human platelets in the presence of von Willebrand factor, nonspecifically precipitates platelet membrane factor VIII antigen. Thus normal platelets contain surface-bound as well as internally stored von Willebrand factor, a protein synthesized by endothelial cells which is necessary for normal platelet function in vivo.


Cell cycle-associated changes in receptors for IgE during growth and differentiation of a rat basophilic leukemia cell line. The rat basophilic leukemia cell line (RBL-1) showed an inverse relationship between growth rate and expression of receptor activity for IgE. After prolonged exponential growth, the number of receptors per cell stabilized at 4-6 times 10-5. Cells in stationary cultures, which are arrested in the G1 phase of the cell cycle, continued to accumulate up to 0.9-1.7 times 10-6 receptors/cell with no increase in volume. Upon resuspension in fresh medium at low density, these cells were shown to lose up to 70% of the receptor activity within 4 h. Assessment of cultures synchronized by double thymidine block and cells fractionated by centrifugation of a Ficoll gradient indicated that the RBL-1 cells acquire receptors in the G1 phase of the cell cycle. No accumulation of active receptors occurred during the S and G2 phases, though the average cell volume increased. Cell division resulted in a drop in number of receptors per cell while the number of cell-bound receptors in the culture remained unchanged. This indicates that during mitosis receptors were simply distributed to daughter cells.


The mechanism of action of the C3b inactivator (conglutinogen-activating factor) on its naturally occurring substrate, the major fragment of the third component of complement (C3b). The fixation of the third component of complement (C3) results in many important biological phenomenon, among which are (a) immune adherence (1), (b) enhancement of phagocytosis (2,3), (c) the release of an anaphylatoxin which is a potent releaser of histamine (4), and (d) the feedback activation of the alternative pathway (5,6). The physiological mechanisms involving C3 fixation require the generation of a C3 convertase which may occur by two separate pathways. C3 convertase can be generated, in the form of C42, by the so-called classical pathway of activation or in the form C3b,B by the alternative or properdin pathway (7). In both cases, C3 is converted to C3b by cleavage of a small peptide, C3a. Normal human serum contains an inactivator of activated C3b. This C2b inactivator or conglutinogen-activating factor (KAF) has been shown to inhibit both immune hemolysis and the immune adherence properties of C3b and to cause cleavage of C3b in the fixed and fluid- phase stages (8-11). Although it is known that the C3b inactivator is not depleted during its reaction with C3b and that C3b treated with the C3b inactivator becomes extremely sensitive to proteolytic digestion by trypsin and "trypsin-like" enzymes (9), the exact molecular nature of the action of the C3b inactivator on C3b has not been studied. In an effort to delineate the products of this interaction, purified C3b and C3b inactivator were allowed to react for various specific lengths of time and the products of these reactions were then analyzed.


Binding of antigen by immunocytes. II. Effect of specific Ig on antigen binding by MOPC 315 cells. The effect of specific immunoglobulin (Ig) on specific binding of antigen to cells has been studied in a model system consisting of nurine myeloma cells (MOPC 315), MOPC 315 serum, and DNP conjugates. MOPC 315 serum, which has IgA specific for DNP, specifically inhibited the binding of DNP conjugates to these cells. Using this model it was found that cells have a marked advantage over free specific Ig in binding multivalent antigen molecules and retaining them in a bound state. Cells were able to specifically bind multivalent antigen in the presence of a large excess of free specific Igm the kinetics of antigen binding to cells was slow, and prolongation of time of incubation increased the amount of specific binding. Both antihapten and anticarrier Ig augmented nonspecific binding of multivalent but not of univalent hapten to control cells. Furthermore, antihapten Ig at low concentration increased antigen binding to specific cells.


Protein requirements of man: comparative nitrogen balance response within the submaintenance-to-maintenance range of intakes of wheat and beef proteins. Sixteen young male students participated in two studies designed to determine the nitrogen balance response to beef or stone-ground, whole wheat protein over the submaintenance-to-maintenance range of protein intake. The objective of the studies was to evaluate the relative capacities of these proteins to meet the minimum protein needs in young adult males. A modified Latin-square design was used to allocate subjects to the four 15-day metabolic balance diet periods in each study. The last 10 days were used for fecal nitrogen determination and the last 5 days for evaluation of urinary nitrogen excretion. The diet periods were separated by a 4-day break period and began with 1 day on a "protein-free" diet. The levels of protien (N times 6.25) intake tested were 0.2, 0.3, 0.4, and 0.5 g/kg body weight/day in the beef study. The results have been compared with those obtained in a previous similar study with egg protein. The regression of estimated "true" nitrogen balance (Y) (mg N/kg/day) on nitrogen intake (X) (mg N/kg/day) was: Y equals 0.51(plus or minus 0.08)X minus 41.9(plus or minus 4.6) for beef and Y equals 0.27(plus or minus 0.06)X minus 33.6(plus or minus 5.0) for whole wheat protein. The amounts of beef and wheat proteins estimated to be requried to support body nitrogen balance in 97.5% of the population supplied 96 and 178 mg N/kg/day, respectively. The relative protein value of beef and wheat proteins, in comparison with egg protein, was 78 plus or minus 12 and 41 plus or minus 10, respectively. It is concluded that the variations in dietary protein quality should be taken into account in assessing the protein adequacy of diets for individuals and population groups.


Diurnal and menstrual cycle variations in serum vitamin B-12 and folacin activity in baboons. Serum levels of vitamin B-12 and folacin were determined every 6 hours over a 24-hour period in five baboons and every other day throughout complete menstrual cycles in seven baboons. All baboons were normally cycling females exhibiting serum estrogen, progestin, and luteinizing hormone levels during the follicular, ovulatory and, luteal phases, respectively, of their menstrual cycles of 217.9 plus or minus 115.6, 420.3 plus or minus 113.9, 207.3 plus or minus 88.6 pg/ml; 1.71 plus or minus 0.84, 2.72 plus or minus 1.50, 5.92 plus or minus 4.38 ng/ml; 2.85 plus or minus 2.3, 38.4 plus or minus 20.5, 6.0 plus or minus 6.6 mg/ml. Serum vitamin B-12 levels did not vary significantly over a 24-hour period although higher mean levels were observed at 1200 and 1800 hours, whereas the lowest mean levels were observed at 2400 and 0600 hours. Folacin activity varied significantly (P smaller than 0.01) over a 24-hour period with the mean level at 1800 hours significantly higher (P smaller than 0.01) than that at all other times. The overall means and standard deviations for vitamin B-12 and folacin, respectively, during this study were 742.2 plus or minus 785.6 pg/ml and 14.48 plus or minus 6.74 ng/ml. Serum levels of vitamin B-12 and folacin did not vary significantly between the follicular or luteal phases or show any significant correlation with the day of the menstrual cycle when analyzed by linear regression procedures. The overall means and standard deviations for vitamin B-12 and folacin, respectively, during the menstrual cycle study were 438.5 plus or minus 137.9 pg/ml and 14.8 plus or minus 4.27 ng/ml.


Perinatal vitamin D metabolism. II. Serial 25-hydroxyvitamin D concentrations in sera of term and premature infants. Serial concentration values of 25-hydroxyvitamin D (25-OHD) were determined in the sera of term, premature, and twin infants. In infants born at term with normal concentrations of 25-OHD in cord blood, serial concentrations of 25-OHD remained normal; in infants born at term with low concentrations of 25-OHD in cord blood, serial concentrations of 25-OHD increased. In premature infants with normal concentrations of 25-OHD in cord blood, serial concentrations of 25-OHD decreased; in premature infants with low values of 25-OHD in cord blood, serial concentrations of 25-OHD did not increase until a postconceptual age of 36 to 38 weeks. The concentrations of 25-OHD in the sera of twin infant pairs followed parallel courses. Oral and intravenous supplementation of vitamin D did not significantly increase the concentrations of 25-OHD in serum of premature infants. These findings suggest that a decreased rate of 25-hydroxylation of vitamin D may be a factor impairing homeostasis of 25-OHD in premature infants.


The erythrocyte sedimentation rate in the newborn period. Determination of the erythrocyte sedimentation rate in capillary blood can be of value in identifying neonates with infection. The normal values for the method described here range from 1 mm/l hour at 12 hours of age to 17 mm/l hour at 14 days of age. Most noninfected neonates with moderate to severe RDS or with other serious illness had values well within the normal range. Infected patients had marked elevations, and the majority of values returned to normal with clinical improvement. Coombs-positive ABO hemolytic disease was also responsible for elevated values. In about half of the infected patients the rise was not seen until 24 to 48 hours after clinical symptoms first appeared. The ESR can be useful in the nursery as a preliminary step in the laboratory evaluation of the sick neonate. Serial determinations may be of aid in identifying the infected infant when the results of bacteriologic cultures are obscured by antibiotic therapy.


Cryopreservation of infective larvae of Dipetalonema viteae. infective larvae of Dipetalonema viteae produced infections in Mongolian jirds (Meriones unguiculatus) after storage of infected ticks (Ornithodoros tartakovskyi) in the presence of dimethyl sulfoxide (DMSO, 5%) for 7 or 595 days in liquid nitrogen (-196 C). Infectivity of these larvae was only partially impaired. Microfilaremias of test jirds were generally lower than those of control jirds given nonfrozen larvae; however, the majority of test jirds developed microfilarial counts suitable for use in infecting ticks. In contradistinction, larvae frozen free of the tick failed to retain infectivity. Apparently the tick, in conjunction with DMSO, protects the larvae during freezing and thawing.


Versatile unit for filling gelatin capsules with drugs or chemicals. A new, inexpensive device was developed for making individually prepared capsules that can be administered to experimental animals. The device was designed for accurate and rapid production of large numbers of various capsule sizes and drug dosages. During 12 months of usage, 21 capsules were prepared weekly for each of 44 dogs (924 total capsules) in approximately 2.5 work-hours/week. Each capsule contained a precise amount of drug to administer a specific dosage to each individual dog. No difficulties were encountered in the manufacture or utilization of this device, and it can be operated by untrained personnel.


[Angiographic study of congenital aortic stenosis (author's transl)]. Left ventriculography is of great interest in the diagnosis and prognosis of congenitalaortic stenosis. It permits one to distinguish valvular stenosis, which is the commonest type, from sub-valvular stenosis, of which there exist 5 main type; type I and II are the commonest, they have a better prognosis than types III and IV. Sub-sigmoid aortography shows the appearance of the sinus of Valsalva, thus whether there exists a tricuspid or bicuspid aortic valve. In all cases, one should seek stenosis at various levels of the aortic outlet and possible myocardiography of the left ventricle, the frequency of which is well known, together with the poor prognosis.


Caffeine-mediated changes of sex ratio in Chinese hamsters, Cricetulus griseus. Sixty 6- to 10-week-old male Chinese hamsters were assigned to two groups. In one, the animals were untreated and in the other, they were treated with 0.02 g caffeine/100 ml water orally for 60 days. The sex ratio of the resulting litters showed a significant (P smaller than 0.025) skewing towards females (61.4%) when compared to that of the controls (49.2%).


Motility of the oviduct and uterus of the cow during the oestrous cycle. Recordings of electrical activity of the oviduct and uterus were obtained during three oestrous cycles in cows fitted with an extra-cellular multi-electrode assembly. The stages of the cycle were identified by the appearance of the cervico-vaginal secretions and changes in the peripheral plasma level of progesterone were determined by radioimmunoassay. A gradual transition from local non-propagating electrical activity to propagating electrical activity with increase in the duration of contractions and then of their amplitude occurred 48 hr before the onset of oestrus. The transition coincided with a rapid decrease in progesterone level from 5 to 10 ng/ml to less than 0-1 to 0-4 ng/ml. This phenomenon was recorded from all uterine electrode sites, but was most marked at the uterotubal junction. Two days before oestrus, trains of potentials and bursts of activity became progressively grouped, apparently randomly, into prolonged phases in the distal portion of the oviduct and over the entire myometrium. During oestrus, the phases of activity became synchronized at these sites and both their amplitude and frequency reached a maximum. The strength but not the frequency of the phases diminished progressively 3 days after oestrus, followed by relative inactivity. The last remaining zone of activity was the uterotubal junction. During oestrus, the activities of the oviduct and the uterus were modified by oxytocin and adrenaline, the effect of the former being more marked on the uterus and that of the latter on the oviduct.


Luteinizing hormone, oestrogen and progesterone levels in peripheral serum of anoestrous and cyclic ewes as determined by radioimmunoassay. Jugular vein blood was collected daily from four mature ewes throughout anoestrus and the first oestrous cycle of the breeding season until 4 days after the second oestrus. The levels of oestrogen, progesterone and LH were determined by radioimmunoassay. There were fluctuations in the LH level throughout most of the observed anoestrous period with a mean plus or minus S.E. value of 2-3 plus or minus 0-9 ng/ml. High LH values of 20-0, 41-2 and 137-5 ng/ml were observed in three ewes on Day - 24 of anoestrus. A brief minor rise in progesterone level was also observed around this period. Progesterone levels were consistently low (0.11 plus or minus 0-01 ng/ml) before Day - 25 of anoestrus. A major rise occurred on Day - 12 of anoestrous and this was followed by patterns similar to those that have been previously reported for the oestrous cycle of the ewe. Random fluctuations of oestrogens deviating from a mean level of 4-40 plus or minus 0-1 pg/ml were observed during anoestrus and the mean level during the period from the first to the second oestrus was 5-2 plus or minus 0-3 pg/ml. A well-defined peak of 13-3 plus or minus 0-7 pg/ml was seen in all ewes on the day of the second oestrus. Results of the present study suggest that episodic releases of LH occur during anoestrus and periods of low luteal activity. The fluctuations in LH levels, as observed during the period of low luteal activity, i.e. before Day - 25 of anoestrus, were less pronounced during the periods of high luteal activity. The view that luteal activity precedes the first behavioural oestrus of the breeding season is supported.


Shifts in ATP synthesis during preimplantation stages of mouse embryos. The actual and potential activities of the cyochrome system were studied in cleavage-stage mouse embroys. Activities were determined by assaying embroys for total ATP and the rates of [32-P]ATP synthesis both before and after their incubation in medium supplemented either with an energy coupling site inhibitor (antimycin, amytal or cyanide) or with the FADH-linked substrate, succinate. The data indicate that there are three major shifts in the mode of ATP production during preimplantation stages: the first, between the two-cell and late four-cell stages; the second, between the eight-celland late morula stages; and the third, between the late morula and late blastocyst stages. These data are discussed in relation to studies on the energy metabolism of cleavage and blastocyst stage mouse embryos.


Effect of prostaglandin F2 alpha on the ultrastructure and function of sheep corpora lutea. Early morphological changes in the ultrastructure of CL of ewes treated with prostaglandin F2alpha were examined in relation to luteal function as judged by plasma progesterone concentration. The luteolytic effect of prostaglandin F2alpha was confirmed, but there was little synchrony between morphological and functional luteolysis. Significant changes included a decrease in the amount of smooth endoplasmic reticulum, a change in the shape of mitochondria and a decrease in the number of membrane-bound granules. There was also an accumulation of lipids.


A comparison of the biochemical activities of Bacteroides corrodens and Eikenella corrodens with those of certain other gramnegative bacteria. Tests carried out on strictly anaerobic and facultatively anaerobic strains of "corrodens" bacteria, showed that although these organisms are relatively inactive biochemically, differentiation can be made on the basis of tests that demonstrate reduction of nitrite, hydrolysis of urea and 1-naphthyl acetate, decarboxylation of lysine and ornithine, and sensitivity to certain selective agents included in culture medium. Plasma was found to be superior to serum in supporting the growth of all "corrodens" bacteria, and a combination of heated and unheated blood added to a nutrient base was shown to yield good growth. Comparative studies are reported with various species of Bacteroides, Haemophilus, Bordetella and related genera.


Temperature dependence of the energy-linked monosaccharide transport across the cell membrane of Rhodotorula gracilis. The temperature dependence of the active monosaccharide transport across the cell membrane of the yeast Rhodotorula gracilis has been studied between 0 and 55 degrees C with D-xylose as the transported substrate: (i) Between 0 and 10 degrees C there is virtually no transport. (ii) The initial velocity of transport increases exponentially from 15 to 30 degrees C (deltaE equal to 32 plus or minus 2 kcal/mol). (iii) At 30 degrees C a sharp "break" occurs in the Arrhenius plot and with increasing temperature the transport becomes inactivated, with a positive slope of the corresponding straight line ("deltaE equal to minus 15 kcal/mol"). (iv) In the temperature range of 50-55 degrees C, both the transport and the metabolic activity cease. In order to account for the abrupt changes of the membrane permeability, we attempted to ascribe them to phase transitions in the membrane structure: the first one, between 10 and 15 degrees C, to the crystalline: liquid-crystalline phase change; the second one, around 30 degrees C, to a change from highly ordered (low entropy) to less ordered (high entropy) membrane structure. Whereas the former phase transition is reversible, the latter appears to be irreversible. Arrhenius plots of the cell respiration exhibit a "break" at 30 degrees C, as well. However, at higher temperatures there is no thermal inactivation of the respiratory activity. The importance of a proper organization of the cell membrane constituents for the efficient transport function is discussed.


Association of race, age, menopausal status, and cerumen type with breast fluid secretion in nonlactating women, as determined by nepple aspiration. Biologic and physiologic factors associated with the availability of breast secretions by a nipple aspiration technique were investigated in 606 normal, non-lactating women. Data obtained on race, age, menstrual status, parity, contraceptive pill use, hormone use, and cerumen type indicated that availability of secretions was related to race, age, and menopause, and to genetic factors associated with apocrine bland function. Fluid aspirates were obtained most often from Caucasians (70.2% success), least often from Chinese (24.1% success). The percentage of successful aspirations declined in women over 50 years of age in all racial groups, but distinctly less so in Caucasians. Chinese and Japanese women with dry-type cerumen had a lower percentage of successful aspirations than those with wet type, which suggested that genetic factors may be associated with breast fluid secretion in nonlactating women.


Histopathology of breast lesions induced in BUF rats of varying ages by ingestion of N-4-(4'-fluorobiphenyl) acetamide. Inbred BUF male and female rats 4, 12, 24, and 52 weeks old ingested 0.04 percent N-4-(4'fluorobiphenyl)-acetamide in a semisynthetic diet for 36 weeks. They were killed 12 weeks later. Susceptibility to mammary carcinogenesis was related to the age and sex of the animals. Younger female rats developed more mammary tumors. Approximately half of these mammary tumors were well-differentiated adenocarcinomas; the remainder were either poorly differentiated or anaplastic adenocarcinomas.


Gas contaminant inhibition of mammary gland differentiation in vitro. A gas mixture of 95 percent 02/5 percent CO2 from several commercial sources was contaminated with a filterable substance (s) that inhibited mammary gland morphologic differentiation in vitro.


Gallium-67 citrate uptake by cultured tumor cells, stimulated by serum transferrin. The uptake of the tumor scanning agent 67-Ga citrate by cultured mouse tumor cells wqs greatly enhanced with small additons of certain sera to the culture medium. This stimulation of uptake was due to a macromolecular serum component, provisionally identified with transferrin. The response under optimum conditions was so great as to suggest that the 67-Ga uptake process depends entirely on the action of transferrin. The same mechanism probably operates in vivo and may help explain interesting features of the tracer's tissue distribution, particularly its tumor affinity.


Lymphocyte antibody interaction in cytotoxicity against human transitional cell carcinoma. The interaction of antibodies and lymphocytes in their immune reaction against human transitional cell carcinomas was studied using the in vitro microcytotoxicity assay. A non-complement dependent, IgG antibody was detected in the serum of occasional transitional cell carcinoma patients, which induced cytotoxicity against transitional cell carcinoma target cells by lymphocytes from donors with and without transitional cell carcinoma. The observation that lymphocytes from transitional cell carcinoma donors were more sensitive to activation by this  anti-transitional cell carcinoma, lymphocyte dependent antibody is compatible with the hypothesis that the surface of lymphocytes from some transitional cell carcinoma donors is coated in vivo with an anti-transitional cell carcinoma lymphocyte dependent antibody and that this antibody may be a significant factor in immunity to transitional cell carcinomas of the urinary tract.


Fricative discrimination in early infancy. In three separate experiments using controlled natural stimuli and a high-amplitude sucking paradigm, infants' ability to detect differences between /s/ and /v/, /s/ and /f/, and /s/ and /z/, respectively, was investigated. Evidence for discrimination was obtained for /s/ versus /v/ and /s/ versus /f/ but not for /s/ versus /z/. Implications for a theory of infant speech perception are discussed.


Speech fluency fluctuations during the menstrual cycle. This study sought to determine whether changes in disfluency characteristics accompany biological and emotional changes during the menstrual cycle. Four extemporaneous speech samples were collected from each of 12 women, one at ovulation (when the average woman experiences her greatest feelings of self-esteem and self-confidence) and one at premenstruation (when she experiences a significant increase in anxiety level) for two consecutive cycles. The mean total frequency of the women's speech disfluencies produced at premenstruation was significantly greater than the mean total of their speech disfluencies produced at ovulation. The disfluency type that accounted for much of the difference in total frequency of disfluency between the two cycle points was revision-incomplete phrase. The tendency for these nonstutterers to produce more speech disfluencies at premenstruation than at ovulation is similar to a tendency we have observed in stutterers.


Management of coarctation of the aorta during pregnancy. In this report, we shall describe the instantaneous electronic monitoring of the fetal heart rate at 24 weeks' gestation. The mother was undergoing resection of coarctation of the aorta. Because the fetal heart rate reflects by physiological and pharmacologic events, monitoring this rate enabled the anesthesiologists and surgeons to make therapeutic intervention sooner and on a more rational basis.


Structure of nuclear pre-mRNA. VI. "Reversed repeats" in animal DNA and their hybridization with double-stranded regions of pre-mRNA. About 2% of mouse DNA reassociates at extremely low Cot values (10-7-10-6 mole-liter-1-sec). This DNA fraction was isolated with the aid of nuclease S1 and purified by chromatography on hydroxyapatite. The study of this fraction suggests that it has a structure of hairpin type. The DNA of the hairpins can hybridize with sequences forming the double-stranded regions in pre-mRNA. Probably at least some of the DNA of the hairpins, described as "reversed repeating sequences" of DNA, is transcribed with the formation of structures of hairpin type in pre-mRNA.


Intragenome distribution of 5-methylcytosine in DNA of healthy and wilt-infected cotton plants (Gossypium hirsutum L.). Fractionation of DNA of healthy and wilt-infected cotton plants has been carried out according to the reassociation kinetics and the content of GC and 5-methylcytosine in the resulting fractions has been studied. The genome of cotton plant was found to be methylated quite unevenly. The GC rich (GC equals 64.7 mole%) fraction of highly reiterated sequences (Cot equals 0-3.7 times 10- minus 2) has a high content of 5-methylcytosine (5.8 mole%), whereas the methylation degree of the fraction of unique sequences (Cot larger than or equal to 487) is very low (the 5-methylcytosine content is about 0.5 mole%). In plants being infected with wilt, the 5-methylcytosine content in DNA or cotton leaves decreases two-fold; no changes in the structure and molecular population of DNA has been found. The sharp change in the 5-methylcytosine content in DNA of infected plants takes place at the expense of the decrease in the 5-methylcytosine content in fractions of highly reiterated sequences. The methylation degree of unique sequences (structural genes) remains unchanged.


A simple nonradioisotope technic for the determination of platelet life-span. Acetylsalicylic acid was shown both in vivo and in vitro to prevent the platelet lipid peroxidation normally induced by the aggregating agents thrombin and epinephrine, and the sulfhydryl inhibitor N-ethylmaleimide. After aspirin ingestion, there was a significant reduction (p smaller than 0.005) in platelet lipid peroxidation, with a gradual return to base-line values over a 10-day period. With these values, a normal platelet survival curve was constructed with a mean half-life of 4.4 days (range of 2.9 to 5.9 days). These values agree with the standard 51-Cr survivals in three patients with chronic idiopathic thrombocytopenic purpura. Half-lives of 1.0, 2.5, and 4.1 days by lipid peroxide technic compared with 1.9, 2.5, and 3.9 days by the concurrent use of 51-Cr. Thus, the technic may be used to measure platelet survival.


Sequelae of central-nervous-system enterovirus infections. The long-term effects of central-nervous-system enterovirus infections were examined in a controlled follow-up study of 19 children 2 1/2 to eight years of age who had been hospitalized with documented enterovirus infection 17 to 67 months before evaluation. Assessment included medical history, physical and neurologic examination, psychologic testing, and speech and hearing evaluation. Three children (16 per cent) had definite neurologic impairment, five (26 per cent) had possible impairment, and 11 (58 per cent) were free of detectable abnormalities. Children whose illness occurred during the first year of life, when compared to controls, were found to have significantly smaller mean head circumference (50.6 vs. 51.6 cm, P less than 0.033), significantly lower mean I.Q. (97 vs 115, P less than 0.007), and depressed languange and speech skills. Children whose illness occurred after the first year of life were not different from their controls. Children whith central-nervous-system enterovirus infection may have neurologic impairment when infection occurs in the first year of life.


Chorioamnionitis and colonization of the newborn infant with genital mycoplasmas. To study the role of Mycoplasma hominis and T-mycoplasmas (Ureaplasma urealyticum) in chorioamnionitis, we obtained culture from 249 puerperal women and their babies. The placentas were examined histologically. Infants whose placentas showed inflammation (chorioamnionitis) had cultures positive for T-mycoplasmas more frequently (37.5 per cent) than those with normal placentas (19.0 per cent) (P = 0.021). Colonization with M. hominis was found in 16.0 per cent of the babies and was not significantly associated with chorioamnionitis. Material colonization with mycoplasmas was more frequent (73.4 per cent) and was not correlated with placental inflammation. We conclude that a substantial proportion of cases of chorioamnionitis may be caused by prenatal infection with T-mycoplasmas. The fact that these organisms are not highly virulent could explain the frequent finding of inflammed placentas from otherwise normal pregnacies. No adverse clinical effects of the placental lesions or of mycoplasmal colonization could be detected in this small study.


Nonpuerperal lactation and normal prolactin regulation. Prolactin secretion was evaluated in 11 consecutive patients referred with nonpuerperal lactation who did not have clinical evidence of pituitary tumors. Six patients had normal fasting prolactin (PRL) levels, 13.8 plus or minus 1.8 ng per ml (Group I), and the other 5 women had elevated basal serum PRL concentrations, 182 plus or minus 72 ng per ml (Group II). All Group II patients had amenorrhea; however, 5 of 6 Group I patients had menstrual periods. The 24-h mean serum PRL concentrations were 12.8 plus or minus 1.2 (SEM), 13.3 plus or minus 0.7, and 165 plus or minus 62 ng per ml for the controls and Group I and II, respectively. A pattern of intermittent PRL discharge during the day characterized each group; however, a normal sleep related increase in serum PRL concentration was absent in the Group II patients. Chlorpromazine produced greater than two-fold increases in serum PRL concentrations in the controls and Group I patients; however, this response was absent in Group II. L-Dopa produced appropriate suppression of serum PRL concentrations in the normals and both patient groups. Normal serum growth hormone, thyroxine, and plasma cortisol characterized each group. Serum estrogen and/or LH and FSH were decreased in 3 of 5 Group II patients; however, the serum concentrations of these hormones were normal in 5 of 6 Group I patients. Short term L-dopa therapy was effective in suppressing lactation in 3 of 5 Group II patients, and it also decreased lactation in the 4 treated Group I patients without significantly altering 24-h mean serum PRL concentrations in the latter group. hypothalamic-pituitary dysfunction was present in certain patients with nonpuerperal lactation who had elevated 24-h mean PRL concentrations and no PRL release following chlorpromazine and sleep. Frequently, however, nonpuerperal lactation is associated with normal prolactin secretion.


Melatonin radioimmunoassay.

Melatonin radioimmunoassay. A radioimmunoassay for melatonin has been developed after the raising of anti-melatonin antibodies in rabbits. The radioimmunoassay is specific and the sensitivity range is greater than the tadpole bioassay.?Author


Opsoclonus: Its clinical value.

Opsoclonus: Its clinical value. Although the exact site of pathologic involvement and the pathogenesis remain uncertain, opsoclonus is an invaluable clinical sign that in it alone may herald an occult systemic malignancy. A thorough systematic evaluation should be undertaken to exclude an occult neoplasm in any patient who presents with opsoclonus unaccompanied by any other clinical sign.


Pharmacologic treatment and evaluation of permanent experimental spinal cord trauma. Permanent paralysis was induced in dogs by a 500 gram centimeter force injury on the spinal cord, and drug treatments were given 1 hour after injury and were continued for 3 days. Dogs were evaluated for 90 days. Isotonic saline or mannitol administration were ineffective in reversing the paralysis. In dogs receiving either dimethyl sulfoxide or dexamethasone, six of eight animals in the former and two of eight in the latter group regained partial or full recovery. The presence of somatosensory evoked responses taken before and at various intervals following trauma showed a good correlation in the prognostic recovery of each animal. It is concluded that dimethyl sulfoxide and dexamethasone can reverse a permanent experimental injury to the spinal cord when given within an hour after trauma.


Bacillus anthracic meningitis.

Bacillus anthracic meningitis. Bacillus anthracis is an uncommon cause of meniningitis. The characteristic hemorrhagic features in two patients who died despite antibiotic treatment are presented and discussed.


X-linked myotubular myopathy with fatal neonatal asphyxia. A second family with X-linked myotubular myopathy is described. The clinical picture includes decreased fetal movements; hydramnios, in at least three cases, probably resulting from insufficient swallowing in utero; and asphyxia at birth. In three autopsy cases many myotubes were found in the muscle tissue. In five definite female carriers, muscle biopsy revealed changes, including myotubes in four. This family probably is not related to the eariler described family with X-linked myotubular myopathy, from which it differs in its 100 percent fatal outcome in the neonatal period, as compared with 25 percent in the eariler described family. A most important finding, in both families, is the possibility of recognizing clinically healthy female carriers by muscle biopsy.


Combined neuroepithelial (colloid) cyst and xanthogranuloma (xanthoma) in the third ventricle. A case is reported of a combined neuroepithelial cyst and xanthogranuloma of the choroid plexus in the third ventricle of a 22-year-old woman. It is suggested that proliferated neuroepithelial cells lining the cyst enter the fibrous wall through the disrupted basal lamina, and then become xanthomatous cells. Disintegration of these foamy epithelial cells releases lipids and other materials into the cyst wall, provoking a response of macrophages and multinucleated giant cells of foreign-body type. A xanthogranuloma is then formed. The origin of "colloid" cysts is from neuroepithelium; these cysts arise from both ependyma and choroid plexus. Those cysts arising in or near the floor of the third ventricle may originate in stomodeal epithelium, but a distinction cannot be made from neuroepithelial cysts by presently available methods.


Study in cerebrovascular disease: brain scanning with technetium 99m pertechnetate; clinical correlations. A prospective brain scanning study using technetium 99m pertechnetate was performed in 84 patients with cerebral infarction. No correlation was found between vascular angiographic findings and the frequency of positive scans. A comparison with clinical motor and mental evaluation showed a good correlation between abnormal scans (58 percent) and poor initial test resutls. A half-life of 17 days characterized the time course of positive scans according to their density and abnormal radioisotopic uptake area.


Cerebrospinal fluid pulse pressure and brain compliance in adult cats. Cerebrospinal fluid (CSF) pulse pressure was inversely proportional to a brain compliance determined from artificial CSF pulsations generated by an external pump. Brain compliance was 0.71 cu mm/mm H2O at steady-state CSF mean pressure and decreased 50 percent for each 240 mm H2O increase in CSF mean pressure (mean results of duplicate experiments on 10 adult cats). Both CSF pulse pressure and the time course of transient changes in CSF mean pressure could be described in terms of a model of the CSF circulation consisting of a constant resistance to CSF absorption in parallel with a pressure-dependent brain compliance.


Does pseudotumor cerebri cause the empty sella syndrome? Eight patients had both the "primary empty sella syndrome," diagnosed by the finding of an air-filled sella turcica at pneumoencephalography, and pseudotumor cerebri, diagnosed by the finding of an elevated cerebrospinal fluid pressure in the presence of normal ventricular size and position on pneumonencephalography. All eight patients were obese women, and six were hypertensive. Six complained of headaches and menstrual irregularities, and two were asymptomatic. Three had visual symptoms and four had papilledema at the time of examination. These two clinical disorders appear to be frequently related, and when they are related, visual field defects and visual loss are more likely to occur than when either entity appears alone. Chronically increased intracranial pressure from pseudotumor cerebri may produce an empty sella if the diaphragma sella is incompetent and the subarachnoid space herniates into the sella turcica.


Bilateral dissecting aneurysms of the intracranial internal carotid arteries in an 8-year-old boy. Nontraumatic intracranial dissecting aneurysms have rarely been reported as the cause of acute infantile and childhood hemiplegia. The present case is unique because dissecting aneurysms occurred bilaterally in two clinically distinct episodes. A recent dissecting aneurysm of the right intracranial internal carotid artery was present with a healed dissecting aneurysm of left internal carotid artery in an 8-year-old boy.


Spontaneous infarction in pituitary tumors: neurologic and therapeutic aspects. In addition to progressive endocrine dysfunction and progressive visual loss, pituitary neoplasms may annouce their presence by the more catastrophic alternative of spontaneous tumor infarction. In two patients reported, illness due to the spontaneous infraction of pituitary tumors was heralded by sudden onset of focal headache associated with diplopia. Stupor, confusion, and evidence of increased intracranial pressure occurred without subarachnoid hemorrhage or massive extrasellar extension of tumor. One patient developed inappropriate antidiuretic hormone secretion with spontaneous infarction in a large but clinically silent chromophobe adenoma. In both patients, skull x-rays suggested a long-standing intrasella mass. Both underwent prompt treatment with endocrinologic replacement therapy and subsequent successful transsphenoidal removal of voluminous, infarcted, pituitary masses.


Unexpected, unexplained death in epileptic patients. Thirty-seven cases of unexpected, unexplained death in epileptic patients were recorded by the Allegheny County Coroner's Office during the years 1969 through 1973. In no case was there anatomic or chemical evidence at autopsy sufficient to explain death. All patients had a duration of epilepsy greater than a year. All but two had less than one seizure per month. Blood levels of anticonvulsants at autopsy revealed only three patients with therapeutic levels of the drugs. Almost 50 percent of the cases studied had no demonstrable anticonvulsant. It is suggested that inadequate levels of anticonvulsant drugs are a significant factor associated with unexpected, unexplained death in epileptic patients.


[Laser-trabecular puncture. Experimental studies]. From the presented evidence it may be seen that a sufficiently strong laser beam can produce performation openings in trabecular meshwork. We were using an exposure time of maximum 200 msec in order not to produce thermal reactions in surrounding tissues. The 1.6 W used by us was sufficient with the above exposure time to produce repeatedly openings in trabecular meshwork. Following this procedure perfusion of Schlemm's canal experiments let the used dye pass through artificially made openings, only slightly staining the trabecular meshwork.


Observations upon the role of the peripheral retina in the execution of eye movements. A variety of experimental situations are described concerning optokinetic nystagmus and tracking movements of the eyes, which have shown that in certain circumstances the peripheral retina can exert a powerful influence upon the resultant response. The teleological significance of the findings is discussed in the context of the mechanisms subserving ocular-following movements.


Malignant tumours of the oral cavity. A study of 177 cases. An unselected material of 177 patients with malignant tumours of the oral cavity is presented and classified according to the TNM system. A therapeutic classification based on the TNM system is proposed. The material is analyzed regarding the 5-year survival rate and the frequency of recurrences in different therapeutic groups.


Neurotological studies upon intoxication by organic mercury compounds. In 1965, a strange illness, which afterwards was diagnosed as the intoxication caused by organic mercury compounds, began to affect the inhabitants of the riverside areas of the Aganogawa River, Niigata, Japan. The authors have investigated the neurotological disturbances, which occurred in 144 cases intoxicated with organic mercurials. The results obtained are summarized as follows: (1) Recruitment (in SISI and other tests) was negative in all the cases except for 32 with inner ear deafness, and Békèsy's AUDIOMETRY (TTS) showed type II: unsettled wide amplitude in 17 cases and severe abnormal adaptation characterizing the type III pattern in nine cases. (2) Spontaneous and positional nystagmus were recorded with ENG in 94 of 144 cases. In 62 of 144 cases, pathological findings of horizontal optokinetic nystagmus were recorded, and those of vertical optokinetic nystagmus were frequently observed in 81 of the 118 cases tested (69 percent). Judging from the authors' observations, neurotological disturbances due to chronic mercurial intoxication may be considered mainly as lesions of the retrocochlea and of the oculomotor system in the brain stem and the cerebellum.


The development of the tympanic sinus.

The development of the tympanic sinus. On the examination of 200 embryonal petrous bones the factors affecting the morphology of the tympanic sinus under physiological conditions are discussed. The development of the tympanic sinus begins during the fourth month of embryonic life. In an embryo of 5 months the floor of the tympanic sinus developing between the cochlea and the vestibulum consists of lamellae resembling a wicker basket. Caudally, its floor reaches at first the lower part of the tympanum, for at this stage the styloid prominence and the pavimentum pyramidis are not yet fully developed. In the eighth and ninth month of foetal life the tympanic sinus is narrowed by the styloid prominence, from the floor and from the cranial side it is bordered by the fully developed ponticulus medialis. In postnatal life the development of the membranous bony substance of the promontory, subiculum promontorii and styloid prominence, each narrows the tympanic sinus in some degree. Under the influence of vascular factors highly developed trabeculae and lamellae are formed in them. The cavity is deeper in the dolichocephalics and is more shallow in brachycephalics.


Artificial stapes - a fourteen-year report. This paper is based on 3,500 cases of otosclerosis operated on during the past 14 years restoring the continuity of the ossicular chain by means of an artificial stapes. Eighty percent showed practically a closure of the air-bone gap. There was no hearing gain in 17 percent of the patients, many of which showed reversal to preoperative levels. Three percent of the patients suffered sensori-neural hearing loss and 1 percent cophosis. Necrosis of the incus occurred in 0.3 percent of the patients and perilymphatic fistula in 0.2 percent. Only one case of inflammatory granuloma was observed as a complication of stapedectomy.


Infections with Eimeria maxima and Eimeria acervulina in the fowl: effect of previous infection with the heterologous organism on oocyst production. Judged by oocyst production, infections with Eimeria acervulina in birds immunized with E. maxima were consistently higher than in birds which had not been immunized.  Oocyst production of E. maxima in birds previously infected with E. acervulina was greater, in three out of four experiments, than in control chickens, but some of the differences were slight.  The findings are discussed but no satisfactory explanation can be offered.  In general there was little or no difference between the oocyst production of the individual species when present as single or mixed infections.


Nosocomial influenza infection as a cause of intercurrent fevers in infants. All patients on an infants' ward manifesting intercurrent fevers were studied for viral and bacterial etiology during a community outbreak of influenza A.  During a one-month period, of 29 infants admitted to the ward, 17 were hospitalized for seven or more days.  Intercurrent fever complicated the course of 13 (76%) of these 17 infants.  Nosocomially acquired influenza A infection was found in 13 infants.  Two of these also contracted a dual infection with influenza B.  The fever lasted an average of 2.7 days with a peak of 38.2 to 39.8 C.  Initial white blood cell counts tended to by high and shifted to the left.  These infants appeared to be at high risk for developing lower respiratory tract disease.  Seven of the 12 had infiltrates on chest X-ray, and five subsequently developed a secondary bacterial pneumonia.  These infants tended to be young, five were under 6 months, and all but one had underlying cardiorespiratory disease.  They also appeared to have prolonge shedding of influenza virus from their nasal secretions.  Six of seven shed the virus for 7 or 21 days.


Cues for screening language disorders in preschool children. This article has described procedures and criteria a pediatrician may employ during office evaluation to identify children suspected of having a developmental language disability.  Disruptions in vocabulary comprehension, vocabulary production, sentence organization, use of word forms, and articulation patterns are important cues signaling the need for referral for a speech and language evaluation.  The 1970 NINDS estimate that no less than one out of every 170 children has a developmental disability affecting the development of language suggests that one or more such children with such a disability enter the pediatrician's office each week.  Many of the negative consequences of this problem may be prevented or greatly reduced by early detection and appropriate referral based upon the informaion described in this article.


[Obstructive cardiomyopathy. A new surgical method of treatment]. A case of treatment of obstructive cardiomyopathy by insertion of a disc mistral prosthesis (Cooley's technique) is reported. The result obtained was satisfactory from a clinical, haemodynamic and angiocardiographic standpoint. The mode of action of this technique is discussed.


[Effect of culture medium components on the biosynthesis of acid ribonuclease by Penicillium brevicompactum cultures]. The effect of different ingredients of the nutrient medium on the synthesis of acid ribonuclease by the culture Penicillium brevi-compactum was studied.  The synthesis of RNase reached maximum on the medium containing glucose, pepton, soya bean flour, ammonium sulphate.  An excessive content of phosphate in the medium decreased the RNase synthesis.  An optimal medium for the synthesis of the enzymes was developed and recommended.


[Formation of ergosterol by yeasts of Candida genus]. The formation of ergosterol by the Candida yeast was studied using glucose, n-paraffins, a mixture of organic acids and inedible sugars.  Yeast of different species and strains showed a similar content of ergosterol which made, as a rule, 0.4--0.5%.  The amount of ergosterol remained relatively stable with varying cultivation conditions and decreased 1.5--2 times with a temperature increase.


[Cultivation of the yeast Mycoderma vini on an ethanol-containing medium and evaluation of the nutritive value of isolated yeast proteins]. A stable process of the development of the yeast Mycoderma vini is possible during continuous cultivation on the nutrient medium containing 3% ethanol as the only carbon and energy source.  The maximum coefficient of ethanol consumption is 65%.  The total protein isolated from the yeast biomass is similar to casein in terms of the composition and susceptibility to gastrointestinal enzymes in vitro.  This protein shows a low content of nucleic acids.