The protonmotive force and beta-galactoside transport in Bacillus acidocaldarius. The acidophilic and thermophilic bacterium, Bacillus acidocaldarius maintains a cytoplasmic pH between 5.85 and 6.31 over a range of external pH from 2.0 to 4.5. Consistently, the pH optimum of beta-galactosidase, as assayed in cell extracts, is between pH 6.0 and 6.5. An electrical potential (delta-psi), interior positive, is also maintained across the membrane. A delta-psi of approximately 34 mV was calculated from determinations of thiocyanate uptake by cells at pH 3.5. Addition of the proton conductor carbonyl cyanide m-chlorophenylhydrazone increased the delta-psi. Treatment of cells with valinomycin (in the absence of external potassium ions) or high concentrations of thiocyanate, to abolish the delta psi, resulted in collapse of the transmembrane proton gradient (delta pH). Active transport of methylthio-beta, D-galactoside occurred optimally at pH 3.5. Transport of the galactoside was inhibited by various compounds which could dissipate the transmembrane delta pH and by respiratory inhibitors. A decrease in the delta pH and an increase in the delta psi occurred upon addition of methylthio-beta, D-galactoside to cells of B. acidocaldarius. Thus the transport of this solute appears to involve an electrogenic symport with protons. The transport system is most active at 50 degrees C and shows little activity at 25 degrees C, although the delta pH is the same at the two temperatures. Gramicidin inhibits methylthio-beta, D-galactoside transport equally effectively at 50 degrees C and 25 degrees C, while nigericin inhibits only after a lag at 25 degrees C.


The effect of pH on the cooperative behavior of aspartate transcarbamylase from Escherichia coli. Saturation curves of activity versus concentration were determined for aspartate transcarbamylase from Escherichia coli (EC 2.1.3.2) for the substrate L-aspartate at saturating carbamyl phosphate (4.8 mM) in buffered solution at pH values from 6.0 to 12.0. Hill coefficients were obtained from the sigmoidal curves. At pH values from 7.8 to 9.1, where substrate inhibition causes difficulties in the Hill approximation, our kinetic scheme includes substrate inhibition and residual activity in the abortive enzyme-substrate complex. The plot of Hill coefficient versus pH has pKalpha values of 7.4 and 9.8 at the half-maximum positions of the curve which has a plateau from pH 8.1 to 9.1. These pKalpha values may be associated with functional groups involved in the allosteric transition which activates the enzyme. A plot of [S]0.5 versus pH shows a pKalpha of 8.5, which may belong to a residue either at or near the aspartate binding site. At 50 mM aspartate concentration the pH-rate profile shows maxima at pH values of 8.8 and 10.0 (cf. Weitzman, P.D.J., and Wilson, I.B.(1966)J. Biol. Chem. 2418 5481-5488, who used 100 mM aspartate). However, when the pH-dependent substrate inhibition is included, the calculated Vmax--H curve is bell-shaped like that of the isolated catalytic subunit.


Indoleamine 2,3-dioxygenase. Purification and some properties. Indoleamine 2,3-dioxygenase was purified from rabbit small intestine to apparent homogeneity as judged by polyacrylamide gel electrophoresis and analytical ultracentrifugation. The native enzyme was a monomeric protein of a molecular weight of 41,000 +/- 1,000 with an s020,w value of 3.45 S. It had a relative abundance of hydrophobic amino acids such as valine, leucine, and isoleucine, and contained approximately 5% carbohydrate by weight. The estimated content of sugar residues per mol of enzyme was: galactose, 1.2; mannose, 2.6; N-acetylglucosamine, 5.2; and sialic acid, 0.8. One mole of enzyme had 0.8 mol of protoheme IX as a prosthetic group. However, copper was not detected in a significant amount and the ratio of copper to heme was less than 0.03. EPR spectra of the nitric oxide complex of the ferrous enzyme indicated that a nitrogen atom, possibly in an imidazole group, might be coordinated as the fifth ligand of the heme coenzyme. The anisotropic g values were gx = 2.08, gy = 1.98, and gz = 2.01. A single enzyme protein catalyzed the oxygenative ring cleavage of D- and L-tryptophan, D- and L-5-hydroxytryptophan, tryptamine, and serotonin. In addition, the purified enzyme had a peroxidase activity with guaiacol and potassium iodide as hydrogen donors, but not a catalase activity.


Isolation of plasma and nuclear membranes of thymocytes. I. Enzymatic composition and ultrastructure. The purpose of this work was to isolate thymocyte plasma membranes at high yield and purity to study specific surface molecules in their structural context. A procedure was developed in which 92-95% of the cells were disrupted by homogenization in a dense viscous medium, while nuclei remained intact. Differential centrifugation of the homogenate was avoided; instead, only a brief (2 h) centrifugation at equilibrium-density of membrane components was used. Five fractions were obtained, three by flotation. Membrane-bound enzymatic activities indicated a 60-80% yield of plasma membranes in the three floated membrane fractions, which comprised 1.6% of the homogenate protein. Enrichment factors for three ectoenzymes, alkaline phosphatase, gamma-glutamyltransferase, and ouabain-sensitive adenosine triphosphatase were respectively, 70-74, and 40-50 in the two lightest fractions. Nuclear membranes were then isolated from the remaining whole nuclei and were found to be enriched in esterase and NADH-cytochrome c reductase. Plasma membranes and light nuclear membranes appeared as pure unit-membrane vesicles in thin sections and freeze-etching electron microscopy. Some aggregation of intramembranous particles occurred in plasma membrane vesicles.


Localization of D-amino acid oxidase on the cell surface of human polymorphonuclear leukocytes. The ultrastructural localization of D-amino acid oxidase (DAO) was studied cytochemically by detecting sites of hydrogen peroxide production in human polymorphonuclear leukocytes (PMNs). Reaction product, which forms when cerous ions react with H2O2 to form an electron-dense precipitate, was demonstrated on the cell surface and within the phagosomes of phagocytically stimulated cells when D-amino acids were provided as substrate. Resting cells showed only slight activity. The competitive inhibitor D,L-2-hydroxybutyrate greatly reduced the D-amino acid-stimulated reaction while KCN did not. The cell surface reaction was abolished by nonpenetrating inhibitors of enzyme activity while that within the phagosome was not eliminated. Dense accumulations of reaction product were formed in cells which phagocytosed Staphylococcus aureus in the absence of exogenous substrate. No reaction product formed with Proteus vulgaris while an intermediate amount formed when Escherichia coli were phagocytosed. Variation in the amount of reaction product with the different bacteria correlated with the levels of D-amino acids in the bacterial cell walls which are available for the DAO of PMNs. An alternative approach utilizing ferricyanide as an electron acceptor was also used. This technique verified the results obtained with the cerium reaction, i.e., the DAO is located in the cell surface and is internalized during phagocytosis and is capable of H2O2 production within the phagosome. The present finding that DAO is localized on the cell surface further supports the concept that the plasma membrane is involved in peroxide formation in PMNs.


A simplified assay of furosemide in plasma and urine by high-pressure liquid chromatography. A simplified high-pressure liquid chromatograhic method for determination of furosemide in plasma and urine has been developed using a fluorometric detector directly coupled to the column effluent. The method includes an ether extraction from acidified biologic samples. The mobile phase used for chromatography on a reversed-phase column (C15 hydrocarbon permanently bonded to silica particles) is sufficiently acidic to induce fluorescence of furosemide. The methylester of furosemide is employed as an internal standard. The sensitivity is 0.1 and 0.25 microgram per ml plasma and urine, respectively. The applicability to pharmacokinetic studies of furosemide is shown.


High-voltage paper electrophoretic assay for guanyl cyclase. A simple, sensitive and rapid technique is described, permitting separation of cGMP from GMP, GDP and GTP by the use of unidirectional high-voltage paper electrophoresis. The recovery of labeled cGMP in the assay of guanyl cyclase, by this procedure is 85-90%; the blank values (no enzyme) are negligible.


Gentamicin-blood agar for isolation of Streptococcus pneumoniae from respiratory secretions. Previous studies have suggested that the yield of Streptococcus pneumoniae from respiratory secretions can be increased by using a 5% sheep blood agar plate supplemented with 5 microgram of gentamicin (GBA) per ml. We report our experience with 245 lower respiratory specimens in which this method was compared with 5% sheep blood agar (SBA) alone. Of 35 specimens with growth of S. pneumoniae on either plate, 21 were detected exclusively on SBA, whereas only 3 were detected on GBA alone (P less than 0.01). By subculturing representative alpha-hemolytic colonies from the final 169 specimens, the yield of S. pneumoniae was increased by 27% compared with the number of identifications that could be made directly from the primary culture. Minimal inhibitory concentrations of gentamicin for the last 25 isolates were greater than or equal to 8 microgram/ml. Our results do not substantiate the previous observations that S. pneumoniae from respiratory secretions gives an increased yield in cultures on GBA.


Diagnosis of pneumococcal pneumonia by antigen detection in sputum. Pneumococcal polysaccharide was detected by counterimmunoelectrophoresis in the sputum of 20 of 26 (77%) adults with community-acquired pneumonia and a positive sputum culture for Streptococcus pneumoniae. The test was negative in 29 pneumonia patients with negative sputum culture for S. pneumoniae. Pneumococcal antigen was also detected in the sputum of six of nine adults with chronic bronchitis and a positive sputum culture, but was not detected in expectorated respiratory secretions of 22 pneumococcal carriers with colds. Pneumococcal antigen could also be detected in sputum by immunodiffusion; antigen titers varied from 1:2 to 1:256. These results strongly suggest that the detection of pneumococcal antigen in respiratory tract secretions indicates infection caused by S. pneumoniae.


Superoxide generation by digitonin-stimulated guinea pig granulocytes. A basis for a continuous assay for monitoring superoxide production and for the study of the activation of the generating system. Stimulation of guinea pig granolocytes by digitonin results in superoxide (O-2) generation. A continuous assay shows that there is a lag between the addition of digitonin and the onset of O-2 production. The rate of activation of the O-2 generating system is dependent upon the concentration of digitonin and the temperature. The final linear rate of O-2 production is affected by the concentration of digitonin, temperature, pH, and the presence of exogenous reduced pyridine nucleotides. Thus, factors which alter either the activation process or the activity of the O-2 generating system can affect O-2 production by stimulated granolocytes.


Inhibition of the bicarbonate exit step in urinary acidification by a disulfonic stilbene. Acidification of the luminal solution by the isolated turtle bladder involves H(+) secretion by a pump at the luminal membrane. The OH(-) dissociated in this process reacts with CO(2) and forms HCO(3) (-) which moves passively out of the cell across the serosal cell membrane. In the present study, this exit step for HCO(3) (-) was inhibited by serosal addition of the disulfonic stilbene, SITS, an agent which is thought to bind to a transport protein at the serosal cell membrane. 90 min after serosal addition of 0.5 mM SITS, H(+) secretion decreased by > 80%. In contrast, luminal addition of SITS had no effect. During inhibition of H(+) secretion by serosal SITS, overall cell pH, measured by the 5, 5-dimethyl-2, 3-oxazolidinedione method, increased from 7.48+/-0.03 to 7.61+/-0.02. This increase of 0.13+/-0.02 pH U was associated with a much larger regional pH increase as judged from the decrement in the attainable pH gradient across the epithelium. After serosal SITS, this gradient was reduced from 2.88+/-0.06 to 2.09+/-0.11 pH U. In the absence of evidence for increased H(+) permeability or a change in the force of the H(+) pump, the gradient decrement of 0.79+/-0.08 U reflects a similar pH increment on the cytoplasmic side of the pump.SITS inhibits the exit of bicarbonate across the serosal cell membrane and, thereby, creates a compartment of high alkalinity in series with the pump. The increased electrochemical gradient across the active transport pathway is the primary factor in the inhibition of urinary acidification.


An androgen binding protein in the testicular cytosol of human testis. Comparison with human plasma testosterone-estrogen binding globulin. After removal of plasma contamination, an androgen binding protein (hABP) was detected in a 105,000-g supernate of human testicular homogenate. The physicochemical properties of hABP have been compared with a similar androgen binding protein in human plasma, testosterone-estrogen binding globulin (TeBG). hABP had high affinity (K(d) = 7.8 nM) and low capacity (0.27 pmol/mg protein) for 5alpha-dihydrotestosterone (DHT). Binding affinity of human TeBG for DHT was greater (K(d) = 0.66 nM, binding capacity 0.68 pmol/mg protein). On the basis of sedimentation rates and Einstein Stokes radii of hABP and TeBG, the mol wt of the two proteins were similar in the range of 87,000-92,000. The ligand specificities of hABP and TeBG were the same. The binding of [(3)H]DHT to hABP and TeBG were reversible processes at 0 degrees C. The half-lives for the dissociation of [(3)H]DHT from hABP and TeBG were 100-120 min and 67-70 min, respectively. Heat sensitivity of hABP and TeBG were similar. hABP had a sharp pH binding curve with an optimum at 8, whereas TeBG had a stable pH optimum between 6.5 and 9. hABP and TeBG were eluted from an ion exchange column at 100 mM and 80 mM sodium chloride concentrations, respectively. Concanavalin A and ricin Sepharose affinity chromatography showed that TeBG is bound to the columns nearly quantitatively, whereas hABP is bound to the columns only partially. Differences between hABP and TeBG, plus the reduction of plasma contamination as marked by albumin, suggest that the human mature testis contains an androgen binding protein separate from that circulating in plasma.


Relationship between phosphorylation of tyrosine aminotransferase and regulation of its synthesis by cyclic AMP and hormones. Tyrosine adminotransferase (EC 2.6.1.5) has been found to be phosphorylated in intact rat hepatoma cells in culture. Incorporation of [32p]i into the enzyme is rapid and is exclusively found as phosphoserine. Cycloheximide treatment reduced phosphorylation of the aminotransferase only slightly and in the presence of three different inducers of this enzyme, dexamethasone, insulin, and dibutyryl cyclic AMP, [32P]I incorporation was increased. It is concluded that [32p]i incorporation into this enzyme probably reflects turnover of phosphate groups associated with pre-existing enzyme molecules catalyzed by a cyclic AMP-independent protein kinase.


The stimulation by catecholamines of guanylate cyclase activity in a cell-free system. Cytosolic guanylate cylase activity in cell-free preparations of the rabbit renal cortex was increased 3- to 5-fold by catecholamines. The plasma membrane-bound enzyme was not activated, although hormone receptors were present. Stimulation was augmented by NaN3, which by itself had little effect on the soluble enzyme activity. With a partially purified enzyme, activity was enhanced by 0.1 muM 1-epinephrine and activated half-maximally by about 1 muM. In decreasing potency, epinephrine greater than isoproterenol greater than norepinephrine greater than dopamine greater than catechol. Phenylephrine and metanephrine did not stimulate. 1-Epinephrine-stimulation of the enzyme was reversed by dialysis and the deactivated enzyme was reactivatable by a second exposure to the catecholamine. Activation by catecholamines was not stereospecific. Epinephrine-stimulated guanylate cyclase activity in the crude cytosolic fraction was partially inhibited by alpha-adrenergic antagonists, but neither alpha- nor beta-blockers inhibited when the partially purified enzyme was used; thus, leaving open the question of a role for typical alpha- or beta-adrenergic mechanisms in this regulation of the soluble enzyme. Adrenochrome was the most potent activator of the partially purified guanylate cyclase, being approximately 10-times more effective than epinephrine. Epinephrine and adrenochrome activated in the presence of reducing agents, i.e., ascorbate, DTT and N2, although the enzyme in a more SH-reduced form and in an oxygen-deficient medium had a decreased sensitivity to both effectors. Epinephrine activated soluble guanylate cyclase in several tissues, including cerebrum, cerebellum, brain stem, lung, heart, liver, ductus deferens and colon. Although the precise mechanism by which low concentrations of catecholamines stimulated guanylate cyclase activity is unknown and the physiological significance of the activation remains to be established, these findings direct attention to an interesting interaction of catecholamines with the cytosolic enzyme system and stress the need for further studies.


gamma-Glutamyl transpeptidase in lactating mammary secretory tissue of cow and rat. Lactating cow and rat tissues were examined for gamma-glutamyl transpeptidase. The specificity of the enzyme for various amino acid acceptors was determined for mammary tissue. The relative specific activity of gamma-glutamyl transpeptidase was high in cow mammary tissue and lower in rat mammary tissue in relation to the activity in kidney of the respective mammals. However, activity per gram wet weight of mammary tissues appeared equivalent. gamma-Glutamyl transpeptidase activity towards specific amino acids agreed with the percent extractions of these amino acids by the mammary gland of the lactating cow in vivo. We believe this is the first evidence of a possible system of amino acid transport by mammary tissues.


Effect of endosulfan on drug metabolizing enzymes and lipid peroxidation in rat. The influence of endosulfan, a chlorinated hydrocarbon insecticide, on rat hepatic drug metabolizing enzymes and lipid peroxidation has been explored in the present study. Total liver weight was significantly increased after endosulfan administration at both dose levels (2.5 and 5.0 mg/kg body weight) without affecting body weight or 9000 x g supernatant protein content. There was a marked increase in the activity of aminopyrine-N-demethylase, aniline hydroxylase, tyrosine amino-transferase and spontaneous lipid peroxidation. In all cases the increase was dose dependent.


Onchocerca sweetae (Nematoda: Filarioidea): notes on the intermediate host. Microfilariae of Onchocerca sweetae are broadly distributed in the superficial layers of the dermis of the water buffalo (Bubalus bubalis). A total of 2855 insects representing 20 species were collected from O. sweetae-infected bait buffaloes. Only one species, Culicoides sp. "M", ingested microfilariae from buffalo skin. Larval development of O. sweetae was observed in the thorax of this species. Atotal of 829 insects, representing 7 species and including 749 parous Culicoides spp. were collected from light and Manitoba traps. Developing filarioid larvae were observed only in Culicoides sp. "M". It is concluded that Culicoides sp. "M" is a natural intermediate host of O. sweetae in the Northern Territory of Australia.


Ultrastructural localization of alkaline phosphatases in rat incisor odontoblasts. The localization of alkaline phosphatases in dentinogenically active rat incisor odontoblasts was studied by means of subcellular fractionation and electron microscopical histochemistry. Subcellular fractionation revealed the predominant phosphatase activity to be present in the microsome fraction and to a lesser extent in the mitochondrial fraction. Adenosine triphosphate degrading enzyme activity was determined in the presence or absence of (+/-)-6(m-bromophenyl)-5, 6-dihydroimidazo(le) (2,1-b) thiazole oxalate (R 8231). Before the histochemical study, the effects on phosphatase activities by aldehyde fixation were studied by biochemical assay. A method of fixation for optimal preservation of phosphatase activity is presented. Phosphatase electron microscopic histochemistry was performed by using ATP as a substrate and with or without addition of the inhibitor R 82319 Precipitates were seen in the membranes of vesicles present in the odontoblast process and the Golgi region. When there were signs of insufficient fixation, precipitates were also seen in the outer membranes of mitochondria. No phosphatase activity was seen in the cell membrane. ATP degrading enzyme activities mediated by nonspecific alkaline phosphatase (APase) and Ca2+ -adenosine triphosphatase thus have the same morphological localization. This close association is consistent with earlier biochemical studies.


Deposition of IgM and complement at the dermoepidermal junction in acute and chronic cutaneous graft-vs-host disease in man. The presence of cutaneous immunoglobulin and complement was investigated in 88 patients with and without graft-vs-host disease (GVHD) after transplantation of bone marrow from HLA identical siblings for the treatment of acute leukemia or aplastic anemia. For comparison, skin biopsies from the patients obtained before transplantation, from 58 healthy individuals (mostly marrow donors) and from four syngeneic marrow recipients were studied. A direct immunfluorescent staining technique was used. Dermo-epidermal IgM deposits were found in 11% of healthy individuals and patients before grafting but were present in 86% of patients with chronic and 39% of patients with acute GVHD. Patients with allogeneic grafts who never had GVHD or who had recovered from it and patients with syngeneic grafts showed findings not different from those in healthy individuals. Findings similar to those with IgM, although less striking, were made for C3, i.e., patients who had chronic or acute GVHD had a high incidence and intensity of C3 deposits at the dermo-epidermal junction. This observation raises the possibility that humoral immunity is involved in the development of GVHD.


Pharmacologic control of the hormonally modulated immune response. III. Prolongation of allogeneic skin graft rejection and prevention of runt disease by a combination of drugs acting on neuroendocrine functions. A recently developed pharmacologic means for suppressing acquired immunity by drugs acting on neuroendocrine regulation has been applied to transplantation immune reactions. A number of drugs have been tested singly and in combination for their capacity to suppress the immune response of mice grafted with allogeneic skin. Another model involved newborn F1 hybrid recipients inoculated with spleen cells from donors of parental strains that had been made specifically "unresponsive" by drug and alloantigen treatment. These procedures led to the identification of a combination of four drugs that induced a remarkable delay in allograft rejection and a prolonged unresponsiveness to alloantigens. This combination of drugs also abrogated the graft-vs-host-runting syndrome in newborn hybrid recipients.


Antibody response to capsular polysaccharide vaccine of Streptococcus pneumoniae in patients with nephrotic syndrome. Twenty-three normal subjects and 19 patients with nephrotic syndrome were vaccinated with tridecavalent pneumococcal capsular polysaccharide vaccine of Streptococcus pneumoniae. The antibody response of the nephrotic patients to pneumococcal capsular antigens was equal to that of the control subjects. These findings indicate that patients with nephrotic syndrome, despite hypogammaglobulinemia, can mount an adequate antibody response to pneumococcal polysaccharides and that there is no evidence of suppressor thymus-derived (T) cells of dysfunctioning bone marrow-derived (B) cells in these patients.


Vulnerability of keto bile acids to alkaline hydrolysis. Rigorous alkaline hydrolysis of the two primary (cholic and chenodeoxycholic) and of the two preponderant secondary (deoxycholic and lithocholic) bile acids found in bile led to excellent recoveries. Such was not the case with 11 different keto bile acid standards. Recoveries for a number of standards were unacceptably low and a variety of artefactural products were tentatively identified by gas-liquid chromatography. Keto bile acids bearing a keto gropu on C-3 were particularly vulnerable. In view of thee findings, quantitative and qualitative data reported on biological specimens submitted to saponification in ethanol, methanol, or even in water are of questionable significance.


Binding of [3H]ouabain to split frog skin: the role of the Na,K-ATPase in the generation of short circuit current. The binding of [3H]ouabain to the serosal side was studied in a chambered preparation of frog skin, free of connective tissue, while the short circuit (Isc) was concurrently monitored. Both ouabain binding and Isc inhibition proceeded as hyperbolic functions of time. A plot of the number of ouabain molecules bound vs. the corresponding values of Isc inhibition (percent) yielded a straight line, yet showed that one-third of the binding occurred before any inhibition of Isc. Upon separation of the skins into two groups based upon initial Isc(Isci)--high, greater than 20 microamperemeter/cm2 and low, less than 10 microamperemeter/cm2, we observed two distinct populations. The high Isci skins bound very little ouabain before inhibition of Isc whereas low Isci skins bound one-half of the total number of sites before exhibiting any inhibition of Isc. These observations strongly suggest that (a) the Na,K-ATPase is directly involved in the generation of Isc, and (b) at low Isc, inhibition of some pumps by ouabain causes a "recruitment" of other pumps to increase their turnover rate and maintain Isc relatively unaffected. In addition, the binding of ouabain also displayed various characteristics that were consistent with known properties of the Na,K-ATPase: (a) increased intracellular K/Na concentrations, whether achieved through the addition of amiloride or removal of Na from the outside medium, led to a significant decrease in ouabain binding rate relative to paired controls; and (b) ouabain binding, either with normal or decreased intracellular Na, was significantly reduced in the presence of elevated K in the serosal bathing medium. Finally, the number of ouabain molecules bound to the frog skins was not correlated with their initial Isc values, indicating that the spontaneous skin-to-skin variation in Isc was not related to the number of functional pump sites but, rather, to their turnover rate.


Preferences by deer mice for solutions of sugars, salts, and acids in Richter-type drinking tests. Deer mice (Peromyscus maniculatus) were tested for taste preferences in 48-hour, Richter-type drinking tests (sapid solution versus distilled water). The Ss, which were seven male and seven female wild adults, were individually housed within an environmental chamber. Test solutions were concentrations of five sugars and three salts (.005 M to 1.0 M) and of two acids (4.0 pH to 1.5 pH). The molar range was too limited to identify the most preferred concentration of glucose, but peak preferences for the other common sugars fell within the range. Preference was shown for hypotonic NaCl concentrations, but other salts and both acids were nonpreferred in all tests. Comparisons were made between these findings and results from Richter-type tests with other murine rodents.


Effect on the Cushing response of different rates of expansion of a supratentorial mass. The effects on the three components (respiration, blood pressure, and heart rate) of the Cushing response (CR) were studied in cats by the continuous expansion of a supratentorial balloon. The rate of expansion was varied over the range of 0.006 to 0.6 ml/min, during whic systemic arterial pressure, heart rate, respiratory rate, and blood gases were monitored. For the different rates the time the CR took to develop, and the balloon volume required for that development were measured. The final volume ("critical volume") for eliciting the CR was more or less constant over the full range of rates of infusion (balloon expansion), a fact that supports that Monro-Kellie doctrine. This constancy of critical volume (CCV) gives rise to a highly statistically significant relationship between the rate of infusion and the latency to the production of the CR, and it is described by a power curve. Thus the development of cerebral dysfunction under these experimental conditions is independent of the rate of expansion and only dependent upon this critical volume. Exceptions to this concept of a critical volume, at the extreme of rates of expansion of lesions in patients, are predicted.


Bacterial etiology of otitis media during the first six weeks of life. Tympanocentesis was performed on 70 infants who had otitis media during the first six weeks of life. The bacteria isolated from their middle-ear effusions were Streptococcus pneumoniae (13 patients), Neisseria catarrhalis (11 patients), Hemophilus influenzae (ten patients), Enterobacteriaceae (four patients), Staphylococcus aureus (four patients), streptococci (groups A and B) (three patients), and Pseudomonas aeruginosa (two patients). Thirty patients (42.9%) had middle-ear effusions which did not contain pathogenic bacteria. Twenty-seven infants were followed for at least 12 months and 12 (44.4%) of these infants had six or more episodes of otitis media during the observation period. Further studies will be needed to establish the significance of middle-ear disease at this age and the role of therapy in improving its outcome.


In vivo and in vitro excystation of Zygocotyle lunata (Trematoda) metacercariae and histochemical observations on the cyst. Encysted metacercariae of Zygocotyle lunata (Trematoda) excyst within 2 hr postexposure in the lower ileum of the domestic chick. Optimal in vitro excystation of this species occurs following pretreatment of the cyst for 15 min in 1% acidified pepsin, treatment in 0.02 M sodium dithionite (a reductant) for 1 to 2 min and then 2 hr treatment in an excystation medium containing 1% sodium glycocholate plus 1% trypsin in Earle's BSS adjusted to pH 8.8 with tris and maintained at 41 C. The cyst of this species is a dome-shaped hemisphere containing an inner and outer wall. The outer wall contains mainly acid mucopolysaccharides, whereas the inner wall is mainly proteinaceous. The cyst contains a ventral lid which only was visualized during excystation.


Studies with Brugia pahangi 17. The anthelmintic effects of diethylcarbamazine. Diethylcarbamazine (DEC) was active in vitro against infective larvae and microfilariae of Brugia pahangi but only at high concentrations. When fed to mosquitoes which were infected with B. pahangi it had little or no activity. In jirds it was inactive against B. pahangi microfilariae and adults when administered at 300 mg/kg for 5 days either by the intraperitoneal or oral route. In cats given 25 or 50 mg DEC/kg intraperitoneally on 3 or 5 occasions it was not microfilaricidal, but most of the adult worms died within 30 days of the end of treatment. Although most microfilariae disappeared from the blood of cats immediately (i.e., within an hour) after treatment, they reappeared within a few hours in the same numbers. Microfilarial levels were reduced after treatment but there was no precipitate decline as occurs in human B. malayi patients.


Tryptamine-induced drug effects insensitive to serotoninergic antagonists: evidence of specific tryptaminergic receptor stimulation? The drug effects of tryptamine and 5-hydroxytryptopham (5-HTP) in the rabbit were compared following monoamine oxidase inhibition and various drug pretreatments. Both agents evoked hyperthermia and behavioural excitation; tryptamine but not 5-HTP also produced forepaw clonic activity. Serotoninergic receptor blockers abolished the effects of 5-HTP but only weakly influenced tryptamine responses. Both tryptamine and 5-HTP effects were potentiated by fluoxetine. Methergoline, a putative tryptaminergic receptor blocker, antagonized tryptamine-induced hyperthermia and forepaw clonus but did not influence 5-HTP responses. It is postulated that while 5-HTP produces its effects through a serotoninergic mechanism, some of the responses to tryptamine result from activation of a specific tryptamine-sensitive mechanism.


Rectal absorption of homatropine [14C]methylbromide in the rat. Homatropine[14C]methylbromide (HMB-14C) was administered to rats by intramuscular injection, oral gavage and rectal suppository. Plasma concentration of 14C were measured over the subsequent 12 h. Peak plasma concentrations were higher and achieved more rapidly after rectal administration than by the other routes whether HMB-14C was administered in a water-soluble suppository base or in aqueous solution. Twelve h after the suppositories were inserted and retained 28% of the 14C had been excreted in the urine while 56% remained in the large intestine. Unlabelled HMB, given in rectal suppositories to anaesthetized rats, caused prompt blockade of the effects of vagal stimulation on pulse rate and of intravenous acetylcholine on blood pressure. These results confirm the rapid rectal absorption of the drug.


The mechanism of the hypothermic effect of amantadine in rats and mice. Amantadine (25--100 mg kg-1, i.p.) given to rats at an ambient temperature of 4 degrees, or mice at 21 degrees, caused a marked fall in rectal temperature. Prior administration of pimozide (1--2 mg kg-1, s.c.) did not block hypothermia due to amantadine in rats or mice; in contrast, hypothermia due to apomorphine (2 mg kg-1, i.p.) and piribedil (10--40 mg kg-1, i.p.) in rats was blocked by pimozide pretreatment. Amphetamine (5 mg kg-1, i.p.) given 2 h after reserpine (2 mg kg-1, i.p.) caused a reversal of the hypothermic effect of reserpine in mice, but a reversal was not obtained with amantadine (50 mg kg-1, i.p.). Direct injection of amantadine (4--8 mg kg-1) into the cerebral ventricles (i.c.v.) of mice caused marked hypothermia which was not blocked by pimozide, but intravenous injection of the same dose of amantadine did not cause hypothermia. Rimantadine, a congener of amantadine but without anti-parkinsonian activity, also caused pimozide insensitive hypothermia in mice at doses of 50 mg kg-1, intraperitoneally or 2--4 mg kg-1, intracerebroventricularly. The main conclusion drawn from these results is that in causing hypothermia amantadine acts in the cns but not on dopamine receptors.


Some pharmacological actions of acetylsecohemicholinium. The effects of the acetylated derivative of HC-3 (acetylsecohemicholinium; AcHC-3) have been studied at cholinergic nerve terminals and compared with the effects of the parent compound. AcHC-3 blocked neuromuscular transmission in nerve-muscle preparations; it was shown to be less effective than HC-3 in producing a pre-junctional block in the rat diaphragm but was more effective than HC-3 in eliciting a post-junctional blocking effect in the chick biventer muscle. On the frog rectus abdominis muscle AcHC-3 caused a substantial potentiation of the contractures elicited by acetylcholine but did not by itself cause a contracture of the muscle. AcHC-3 inhibited the synthesis of acetylcholine by cholinergic nerve ending particles and inhibited the uptake of [14C]choline into brain synaptosomal fractions to a similar extent to HC-3. AcHC-3 was shown to be a substrate for cholinesterase enzymes although the rate of hydrolysis was much less than the rate of hydrolysis of acetylcholine. It is concluded that AcHC-3 is effective in inhibiting cholinergic transmission and this action is exerted by the open chain (seco) compound and is not due to the hydrolysis of the AcHC-3 by cholinesterases to form the active HC-3 molecule.


A mammalian muscle with the pharmacological characteristics of slow tonic muscle. The guinea-pig isolated cremaster muscle gave tetrodotoxin-resistant dose-related contractures with acetylcholine, carbachol and the depolarizing neuromuscular blocking agents. The dose-response curve to suxamethonium in tetrodotoxin 2 X 10(-7)M could be shifted to the right with tubocurarine 10(-6)M. KCl, 0.1M, produced slow sustained contractures of the muscle. With the cremaster nerve-muscle preparation tetanic contractions at 20 Hz were maintained over several minutes. Tetrodotoxin eliminated the twitch response to single shock nerve stimulation but not the sustained increase in tension produced by suxamethonium. The results suggest either that there is a component of slow tonic muscle in the guinea-pig cremaster or that the cremaster consists of a type of focally innervated muscle which has pharmacological responses qualitatively different from those of most focally innervated muscles so far described.


Effect of carbenoxolone on lipolysis in rat adipose tissue. Carbenoxolone slightly but significantly decreased the release of FFA from rat epididymal fat pads. The antilipolytic action of carbenoxolone was not blocked by 10(-3)M 3-isobutyl-1-methylxanthine, a potent inhibitor of phosphodiesterase. The findings suggest that carbenoxolone exerts its antilipolytic activity by acting on adenylate cyclase, thereby decreasing cyclic AMP concentrations and the activity of the hormone-sensitive lipase in adipose tissue.


The inhibition by clorgyline of 5-hydroxytryptamine deamination by the rat liver. The inhibition of the monoamine oxidase activity in the rat liver by the substrate selective inhibitor clorgyline has been investigated with 5-hydroxytryptamine as substrate. The results obtained are consistent with a theoretical model whereby the inhibition of enzyme activity by clorgyline follows a reversible association phase leading to an irreversible 'suicide' reaction. The relative concentrations of enzyme and inhibitor are of the same order, and can account both for the failure of the reaction to go to completion, and for the differences in the apparent sensitivity of enzyme preparations to inhibition by clorgyline. The possible value of this type of inhibition as a means of assay for monoamine oxidase active centres is discussed.


Interaction of cortisol-21-palmitate with liposomes examined by differential scanning calorimetry. Liposomes have been suggested as carriers for corticosteroids in the local treatment of arthritis by intra-articular injection. The long chain 21-esters of cortisol such as the palmitate or octanoate are taken up and retained by liposomes in higher concentration than cortisol itself. Differential scanning calorimetry has been used to show that the cortisol ester is anchored in the liposome phospholipid bilayer by the acyl side chain. In addition, the limiting concentration of cortisol-21-palmitate which can be incorporated into dipalmitolyl phosphatidylcholine liposomes has been measured by observing changes in the DSC spectrum at different steroid concentrations. Steroid in excess of this concentration limit forms a separate phase which can be identified by nuclear magnetic resonance. For optimum effect, the treatment of arthritis with liposomes must be carried out with liposomes containing steroid below the limiting concentration.


The measurement of the adhesion of film coatings to tablet surfaces: the effect of tablet porosity, surface roughness and film thickness. The effect of tablet porosity, surface roughness and film thickness on the adhesion of hydroxypropyl methyl cellulose films to placebo tablet substrates have been studied using a specially designed tensile tester (Fisher & Rowe, 4976). There were direct relations between measured adhesion and tablet porosity and also surface roughness and tablet porosity. The effect of film thickness on the measured adhesion is complex with an initial decrease with thicknesses up to 35 micron and then a gradual increase with thicknesses up to 140 micron dut to differences in the strees distribution within the film during testing. A knowledge of these effects is necessary if results from various sources are to be compared. The findings illustrate the potential capability of the extrapolation of measured adhesion results to zero porosity and zero thickness values in order to obtain a measure of the true or intrinsic adhesion at any film/tablet interface without the confusing elements of tablet porosity, surface roughness and residual stresses in the film.


On methods of expressing dissolution rate data. The value of the Weibull, logarithmic-logistic, and logarithmic-normal plots in expressing dissolution rate data is considered for the combinations of zero and first-order release with sink and non-sink conditions, and for actual dissolution rate data of diazepam from tablets in a medium of pH2.


Correlation of physicochemical properties with absorption and metabolism of some tricyclic drugs. Octanol and dodecane partition coefficients, surface activity and adsorbability to activated charcoal were determined for six tricyclic psychotropic drugs with N-dimethylalkyl side chains. Surface activity correlated well with the partition coefficients, and all drugs obeyed the Langmuir adsorption isotherm. A correlation between the reciprocal of the death time of gold fish exposed to drugs and partition coefficients was observed. The extent to which the drugs were N-demethylated as measured by formaldehyde formed in rat liver homogenate incubations correlated with their adsorbability to activated charcoal but not with their ability to inhibit aniline-p-hydroxylase, nor was there a linear correspondence between N-DEMETHYLATION AND DRUG LIPOPHILICITY AS INDICATed by partition coefficients or surface activity.


Investigation of possible interactions between substance P and transmitter mechanisms in the substantia nigra and corpus striatum of the rat. The effect of substnace P (SP) on the uptake and release of radiolabelled dopamine (3H-DA), 5-hydroxytryptamine (3H-5-HT) and y-aminobutyric acid (3H-GABA) was studied in slices of rat substantia nigra and corpus triatum. SP, 10(-9) to 10(-5) m, failed to modify the uptakes of these compounds during incubations (10-90 min) with slices of either brain region. SP, 10(-6)M, had no apparent effect on the spontaneous output of any of these compounds in either substantia nigra or corpus striatum. In the corpus striatum, SP seemed to potentiate the potassium-stimulated outflow of 3H-DA and 3H-5-HT, but not 3H-GABA, while the realeases from substantia nigra were unaffected. Morphine (10(-3)M), but not met-enkephalin (5 X 10(-6)M), weakly antagonized K+- EVOKED RELEASE OF 3/-DA in the corpus striatum. These results are discussed with reference to the possible interaction of SP with transmitter mechanisms at presynaptic sites in the central nervous system.


Quisqualamine, a novel gamma-aminobutyric acid (GABA) related depressant amino acid. A new substnace, quisqualamine, the decarboxylated analogue of quisqualic acid, predictably depressed electrical activity of neurons of the frog and rat spinal cord in vitro and of the mouse spinal cord in vivo. In the in vitro preparations, the action of quisqualamine was associated with a prolonged depolarization of primary afferent terminals which was sensitive to blockade by picrotoxin and bicuculline and which was also depressed by strychnine. This suggests an interaction of quisqualamine with presynaptic receptors for both GABA and beta-alanine. Post-synaptic actions of quisqualamine, which were less marked than those at presynaptic sites, also appeared to be predominantly GABA-mimetic in vitro, though a sensitivity to the GABA-antagonist bicuculline could not be demonstrated in vitro.


Rat brain and serum lithium concentrations after acute injections of lithium carbonate and orotate. Eight hours after intraperitoneal injections of 1.0, 2.0, and 4.0m equiv Li kg-1, the serum and brain lithium concentrations of rats were significantly greater after lithium orotate than after lithium carbonate. While little serum lithium remained at 24 h after injection of 2.0 m equiv kg-1 lithium carbonate, two-thirds of the 2 h serum lithium concentration was present 24h after lithium orotate. Furthermore, the 24 h brain concentration of lithium after lithium orotate was approximately three times greater than that after lithium carbonate. These data suggest the possibility that lower doses of lithium orotate than lithium carbonate may achieve therapeutic brain lithium concentrations and relatively stable serum concentrations.


Corticosterone concentrations in mice during ethanol drinking and withdrawal. Consumption of an ethanol-containing diet by mice resulted in a significant increase in circulating concentrations of corticosterone which was maintained for 8 days. There were no changes in the concentrations of plasma corticosterone binding globulin. Ethanol withdrawal symptoms followed the removal of ethanol from the diet and circulating corticosterone concentrations were further increased. There was no correlation between blood ethanol and glucocorticoid concentrations during the chronic ethanol treatment. Stress related to ethanol consumption may be of greater importance than the circulating ethanol concentrations in producing the elevation in plasma glucocorticoids.


Antidotal effects of dimethyl sulphoxide against paracetamol-, bromobenzene-, and thioacetamide-induced hepatotoxicity. In mice the hepatotoxic effects of paracetamol (0.5-1.0 g kg-1, orally) as evidenced by increased serum enzyme activities of the aminotransferases and sorbitol dehydrogenase were dose-dependently inhibited by simultaneous treatment with dimethyl sulphoxide (DMSO 0,25-1.0 g kg-1, i.p.). DMSO was also active against bromobenzene- and thioacetamide-induced hepatotoxicity, but failed to protect mice against carbon tetrachloride-induced liver damage. Hepatic glutathione depletion in mice amounting to 94% after paracetamol (0.5 g kg-1, orally) and to 60% after bromobezene (0.25 ml kg-1, orally) was dose-dependently reduced by the simultaneous administration of DMSO(0.25--1.0 G KG-1, I.P.). This indicates less conjugation of the toxic metabolites of paracetamol and bromobenzene to liver glutathione (G-SH) in the presence of DMSO.


Rearrangement of N-benzyl-2-cyano-2-(hydroxyimino)acetamide. The reduction of N-benzyl-2-cyano-2-(hydroxyimino)acetamide resulted in the formation of N-benzyl-1,2-ethanediamine and N-benzyl-N'-methyl-1,2-ethanediamine in approximately an equimolar ratio. The formation of the two unexpected products is explained by the migration of a cyano group in a Beckmann-type rearrangement.


Interaction of tricyclic antipsychotic and antidepressant drugs with 1-anilino-8-naphthalenesulfonic acid. The binding of 1-anilino-8-naphthalenesulfonic acid to selected tricyclic antipsychotic and antidepressant drugs was studied by fluorescence spectroscopy. The acid exhibited an increase in fluorescence intensity accompanied by a hypsochromic shift of the emission lambdamax in the presence of these drugs. These fluorescence characteristics, in addition to those of acid-drug complexes after addition of potassium chloride or urea, suggested that binding was hydrophobic. The spectra also provided evidence regarding the importance of certain structural features of drugs in determining the nature of binding.


Solubility studies of silver sulfonamides. The solubilities of silver sulfapyridine, silver sulfamethazine, and silver sulfamethizole as a function of pH were determined in nitric acid-potassium nitrate, acetate, and sulfonic acid buffers. All silver sulfonamides showed an increase in solubility with increasing hydrogen-ion concentration, a behavior which closely paralleled the protonation of the p-amino function of the sulfonamide. A silver-ion selective electrode was used to measure silver-ion concentration in solution and the methods of known subtraction and known addition were used to measure total silver. Both silver sulfamethizole and silver sulfamethazine were ionized completely in solution. Silver sulfapyridine was ionized completely only in the more acidic pH 2-3 range. A comparison of the physical properties of the silver salts for which mortality studies were available revealed a unique set of properties for silver sulfadiazine.


Ketonic oxidation products of cyclobarbital. Chemical or biochemical oxidation of cyclobarbital yielded 5-(3-oxo-1-cyclohexen-1-yl)-5-ethyl -2,4,6,- (1H,3H,5H)-pyrimidinetrione, and photochemical oxidation gave 5-(6-oxo-1-cyclohexen-1-yl)-5-ethyl-2,4,6-(1H,3H,3H)-pyrimidinetrione. These compounds were isolated and purified by TLC, and their structures were determined by UV, IR, NMR, and mass spectral data; the crystalline structure was determined by X-ray diffraction.


Colorimetric determination of nadolol in tablets. Nadolol was extracted from tablet excipients with an acidic potassium chloride solution. The drug was oxidized with periodic acid, and the resulting aldehyde was reacted with 2,4-dinitrophenylhydrazine to form the corresponding hydrazone. Excess reagent was removed with a cupric chloride solution. The hydrazone was extracted into chloroform, and its absorbance was measured at the 352-nm maximum.


Possible ion-pair-mediated absorption of mixidine I: partitioning and lethality studies. Mixidine, a very soluble base which is completely ionized in all physiological fluids, was found to form ion-pairs as demonstrated by its ability to partition into 1-butanol from acidic solutions. A similar relationship was observed for the effect of acids on the absorption of intraduodenally and orally administered solutions of mixidine in rats (as determined by lethality). Studies also demonstrated that the pH-lethality effects were not specific for a particular counterion. Mixidine was more lethal when administered intraduodenally than when administered orally, and the counterions per se were not lethal in the doses used.


Possible ion-pair-mediated absorption of mixidine II: plasma levels and histology. Plasma intact 14C-mixidine levels in rats increased when the drug was administered intraduodenally with 1:3 and 1:5 molar ratios of 2-naphthalenesulfonic acid. Upon histological examination of the duodenums, similar doses of mixidine combined with 2-naphthalenesulfonic acid produced no dose-related lesions. These and previous observations demonstrate that mixidine absorption may be enhanced by ion-pair formation.


Simple GLC analysis of anticonvulsant drugs in commercial dosage forms. A simple, specific GLC procedure is described for the analysis of one sedative and six anticonvulsant drugs in pharmaceutical dosage forms. Sample aliquots of ethotoin, glutethimide, mephenytoin, methsuximide, and phensuximide were shaken with or extracted into ethyl acetate, diluted with the internal standard (diphenyl phthalate) solution, injected into a gas chromatograph, and eluted from a methylsilicon column. Primidone and phenytoin samples (extracted as the free acid) required derivatization with N,O-bis(trimethylsilyl)acetamide prior to chromatography. The same temperature programming conditions and flow rate settings were used for all seven drugs. The GLC results agreed well with those obtained using the pharmacopeial methods.


Effects of patulin on the kinetics of substrate and cationic ligand activation of adenosine triphosphatase in mouse brain. Patulin (4-hydroxy-4H-furo[3,2-c]pyran-2(6H)-one), a carcinogenic lactone produced as a major metabolite by several fungi, inhibited the Mg++-dependent Na+-K+ activated adenosine triphosphatase (ATPase) activity of mouse brain microsomal fractions with an estimated IC50 of 3.0 X 10(-4) M. Inhibition was concentration dependent. Hydrolysis of ATP was linear with both time and enzyme concentration either with or without patulin in reaction mixtures. Altered pH and activity curves for Na+-K+ ATPase demonstrated comparable inhibition by patulin in buffered acidic ranges through an optimum of 7.5, followed by a reduction of toxicity to this system at higher alkaline pH. Kinetic studies of cationic-substrate activation of Na+-K+ ATPase indicated noncompetitive inhibition with respect to ATP (at low affinity nucleotide-directed sites) and Na+ (in the presence of low, noninterfering concentrations of K+). Competitive inhibition with respect to activation of the Na+-k+-stimulated activity and K+-stimulated p-nitrophenyl phosphatase activity of the enzyme system was indicated by altered binding site parameters without change in apparent Vmax in the presence of patulin. Activity was partially restored by washing. Preincubation of patulin with dithiothreitol or glutathione protected the enzyme from inhibition. Results suggest that patulin exerted its effect on Na+-K+ ATPase either directly by interfering with K+ binding or indirectly by inducing a conformational change in the enzyme.


Beta adrenergic blockade, regional left ventricular blood flow and ST-segment elevation in canine experimental myocardial ischemia. The effects of dl-propranolol, d-propranolol, dl-pindolol and dl-practolol on regional myocardial blood flow (assessed by means of tracer microspheres) and on ST-segment elevation in ischemic and nonischemic areas of the canine left ventricle have been investigated. dl-Propranolol and dl-pindolol, but not dl-practolol and d-propranolol, induced blood flow redistribution from the epicardium to the endocardium both in ischemic and nonischemic areas. dl-Propranolol-induced redistribution was abolished by atrial pacing at the control heart rate value. These results indicate that the redistribution phenomenon only occurs if both a bradycardia-inducing beta1 adrenoreceptor blockade and a coronary vessels beta2 adrenoceptor blockade are simultaneously achieved. All four drugs significantly decreased ST-segment elevation in ischemic areas. Under atrial pacing, this effect was abolished with dl-practolol but only reduced with dl- and d-propranol, suggesting that, besides bradycardia, membrane stabilization might be involved in protection against ST-segment elevation in ischemic areas.


Liberation of cyanide from alpha-aminonitriles relative to amygdalin. A series of aminoitriles have been synthesized and studied whose nonenzymatic dissociation with release of cyanide may be varied by modest alteration of their molecular structure from that obtained with nonenzymatic dissociation of amygdalin to that obtained from enzymatic dissociation of amygdalin by substantial quantities of beta-glucosidase. The relationship between such alterations in molecular structure and nonenzymatic dissociation is discussed. A combination of the results of these studies and studies relating molecular structure to physical localization propensity in tumors has potential in the design of chemotherapeutic agents.


Inhibition of sympathetic nervous system by histamine:studies with H1- and H2-receptor antagonists. A study was designed to investigate the effect of histamine on sympathetic transmission to the myocardium in pentobarbital-anesthetized dogs. Intravenous infusion of histamine (1.0 and 2.0 microgram/kg/min) produced dose-related decreases in blood pressure and caused significant impairment of cardioacceleration observed during stimulation of the right postganglionic cardiac sympathetic nerve fibers. Positive chronotropic effects of intravenously administered norepinephrine as well as tyramine were not altered during histamine infusion. Blockade of neuronal reuptake with desipramine did not modify the inhibitory action of histamine on sympathetic nerve function. Prior administration of mepyramine (pyrilamine), a histamine type H1-receptor antagonist caused partial attenuation of the depressor action of histamine, but did not prevent histamine-induced inhibition of neurogenic function. Further treatment with metiamide, a histamine type H2-receptor antagonist caused nearly complete attenuation of the depressor response to histamine and also significantly antagonized the inhibitory action of histamine on sympathetic transmission to the myocardium. It is concluded that while the depressor action of histamine is due to the activation of both H1- as well as H2-receptors, histamine causes impairment of sympathetic nerve function to the myocardium by acting on H2-receptors which may be located on sympathetic nerve terminals.


The role of peripheral catecholamines in oxotremorine tremor in the rat and its antagonism by beta adrenoceptor blocking agents. Oxotremorine, 0.25 mg/kg, produces marked tremor in the rat, which is abolished by scopolamine, 0.5 mg/kg, and is substantially reduced in intensity and duration both by adrenalmedullectomy and by chemical sympathectomy with 6-hydroxydopamine. Oxotremorine increases plasma norepinephrine from 0.62 +/- 0.07 to 3.01 +/- 0.47 ng/ml and plasma epinephrine, from 0.82 +/- 0.14 to 3.42 +/- 0.48 ng/ml, in conscious unrestrained rats. l-Propranolol (0.5-2.5 mg/kg) reduces tremor, and at 2.5 mg/kg is more effective than either chemical sympathectomy or adrenal demedullation. d-Propranolol and sotalol are also active at 4 and 10 times the dose of l-propranolol, respectively. l-Propranolol does not prevent the rise in catecholamines induced by oxotremorine. It is suggested that stimulation of central muscarinic receptors causes tremor by a combination of two effects. There is an increase in cholinergic influence to motor efferents accompanied by an activation of the sympathoadrenal system to release catecholamines which augment tremor by stimulation of beta2 adrenoceptors.


Synthesis and pharmacological activity and some derivatives of 1-phenyl-1,2,3,4-tetrahydro-5H-1,4-benzodiazepin-5-one. 4-N-Alkylamino derivatives and corresponding ammonium quaternary salts of tetrahydro-1,4-benzodiazepin-5-one were synthesized and evaluated for psychotropic activity in mice by ip via. This study was also extended to some nitro and amino derivatives of tetrahydro-1,4-benzodiazepin-5-one. Compounds were devoid of tranquilizing activity and in comparison with two classical benzodiazepines, chlordiazepoxide and diazepam, they showed high toxicity and little or no effect on motor coordination, motor activity, and maximal electroshock. On some "in vitro" tests the compounds exhibited pharmacological properties when they were used at high concentrations.


A method for the isolation of Bacteroides melaninogenicus from the human mouth. Isolation procedures involving the direct plating of specimens and the use of selective, non-selective, transport and enrichment media were compared in respect of their value for the recovery of Bacteroides melaninogenicus from the gingival sulcus. A selective medium containing kanamycin and vancomycin enhanced the recovery rate of B. melaninogenicus. VMGII transport medium was convenient to use and gave as good a recovery rate as that obtained with plates directly inoculated in the clinic.


Amino acid activation with adenosine 5'-phosphorimidazolide. Amino acids are activated by reaction with adenosine 5'-phosphorimidazolide in aqueous imidazole buffers. If adenosine 5'-(O-methylphosphate), an analogue of the 3'-terminus of t-RNA is present, 2'(3')-O-aminoacyladenosine 5'-(O-methylphosphate) is formed. Fifteen percent of this compound accumulated at pH 5.8, but less was formed at higher pHs. The highest efficiency of utilization if ImpA attained in our experiments was about 24%. Analogous reactions occured with several other amino acids, including a number that have functional side-chains.


Intracellular electrolytes in erythrocytes during and after shock: relation to impaired consciousness. Of 32 patients in shock and catabolic state following shock with impaired consciousness 31 exhibited raised sodium content in their erythrocytes. On the assumption that the erythrocyte membrane acts the same as nerve cell membrane, the hypofunction of erythrocyte membrane may result in impaired consciousness. The hypofunction of erythrocyte membrane is assumed by its increased permeability in shock. A positive osmotic discrepancy between measured and calculated levels denotes altered membranous permeability. Subjectively, impaired consciousness was evaluated by clinical grading. Meanwhile, as a trial of quantification of conscious levels, we applied a new technique of analysis of power spectrum obtained by computer on the autocorrelogram of the EEG during intermittent photic stimulation. This new analytic method was useful in evaluating objective changes of cerebral function. There was a good correlation between raised sodium content in erythrocytes and depressed power spectrum. The degree of increased sodium in erythrocytes seems to correlate with patients' clinical prognosis.


Transfection of Streptococcus pneumoniae with bacteriophage DNA. It was possible to transfect Streptococcus pneumoniae with DNA obtained from a newly isolated bacteriophage, diplophage-4 (Dp-4). Optimal frequency of transfection (0.9%) required the use of a nuclease-defective mutant; with wild-type bacteria, the transfection frequency was about 100-fold lower. Transfection requires physiological conditions that appear to be similar to the competent state needed for genetic transformation (A. Tomasz, J. Bacteriol. 91:1050--1061, 1966).


Helper-independent transformation by unintegrated Harvey sarcoma virus DNA. We have studied the unintegrated infectious DNA of Harvey sarcoma virus (Ha-SV) and Moloney leukemia virus (Mo-MuLV). The source of infectious viral DNA was the Hirt supernatant fraction from cells acutely infected with Ha-SV and Mo-MuLV. To obtain a direct quantitative assay for infectious viral DNA, recipient mouse cells were first exposed to calcium phosphate-precipitated viral DNA and then treated with dimethyl sulfoxide. Infectivity was monitored by focus formation for Ha-SV and XC plaque formation for Mo-MuLV. The viral DNA titration pattern followed single-hit kinetics for both foci and plaques, indicating that a single molecule carried information for each function. Focus-forming and plaque-forming activity were present in different molecules, since these two biological activities could be separated from each other by agarose gel electrophoresis. The focus-forming molecule was linear DNA with a molecular weight of about 4 x 10(6) daltons. The focus-forming activity of the viral DNA was sensitive to EcoRI and resistant to XhoI restriction endonucleases, whereas the plaque-forming activity was resistant to EcoRI and sensitive to XhoI. The generation of helper-independent foci indicates that Ha-SV DNA can transform mouse cells in the absence of helper virus or its proteins.


Lymphangiography in patients with malignancy in a non-descended testicle. Lymphangiography was performed on 23 patients with malignancy in non-descended testicles, 14 of whom had seminomatous and 9 non-seminomatous tumors. Lymph node metastases were shown by lymphangiography in 8 patients: 3 in lumbar nodes, 1 in iliac nodes alone and 4 in lumbar and iliac lymph nodes. Microscopic metastases were shown in lumbar nodes at retroperitoneal lymph node dissection in 2 patients when the lymphangiograms were negative. Iliac lymph node metastases are rare in testicular tumors but may be seen in tumors of non-descended testicles, alone or in combination with lumbar metastatic disease. This information is extremely important for the radiologist as well as the clinician in the management of patients.


Screening urography in asymptomatic cryptorchid patients. Excretory urograms in 140 cryptorchid patients free of associated urologic symptoms are reviewed. Our findings are presented and compared to those of previously reported series. Our results do not indicate a specific indication for the routine use of excretory urography in asymptomatic male subjects with cryptorchidism.


Anorchism versus cryptorchidism: the importance of a diligent search for intra-abdominal testes. We reviewed 13 cases of testicular tumors arising in cryptorchid, intra-abdominal testes. All types of germinal neoplasms were found, seminoma being the most common. Of the 13 patients 5 had undergone limited exploration previously, with no testis being identified and, thus, were thought to have had unilateral anorchism. The importance of a thorough exploration of the peritoneal cavity is emphasized.


Prostatic utricle cysts (müllerian duct cysts). A series of 88 male patients with prostatic utricle cysts (müllerian duct) has been compiled by adding our 3 patients to 85 cases reported. Irritative lower urinary tract symptoms were the most common presenting complaint (42 per cent), while lower obstructive symptoms were noted in 29 per cent. Of the 20 clinical case reports in boys 25 per cent presented with epididymitis often in an undescended gonad. A cystic rectal mass found in half of the patients represents the most common presenting sign, while hypospadias occurred in a fourth. Unilateral renal dysgenesis or agenesis was associated in 10 per cent of the reports. While the suprapubic surgical approach to excision has been used most commonly it has proved unsatisfactory in two-thirds of the cases. The posterior approach was used successfully in 2 boys with preservation of erectile potency in both. The wide spectrum of histopathology of these cysts prevents clear characterization. A 3% incidence of malignancy in prostatic utricle cysts is noted.


Reconstruction of the urinary tract in prune belly uropathy. Ten patients with prune bell uropathy, including 7 neonates, were treated with extensive surgical tailoring of the upper urinary tracts using primarily the upper ureteral segment. Simultaneous transabdominal orchiopexy, particularly in the neonate, is described as a useful adjunct. Our early results in these patients justify this aggressive approach.


Treatment of the agitated patient with an organic brain disorder. Agitated patients with organic brain disorders represent relatively common diagnostic and management problems. Therapeutic failures usually result from a failure to understand the patient's disturbed behavior, the staff's emotional response to the patient's behavior, or neglect of the biological cause and ineffective use of medication. Effective management depends on continued monitoring of the patient's mental status and physical condition.


[Comparative examinations of gastric juice in healthy infants and in infants suffering from enteritis (author's transl)]. There are significant differences between the shapes of curves of gastric juice volume, acid quantity, pH levels, and peptic activity, in the three age groups of healthy infants under examination. Significant differences are also seen in all three age groups with regard to the quantity of acid calculated in relation to 1 ml gastric juice, as well as the relative peptic activity (volume activity). The amount of gastric juice is proportional to the bodyweight and body surface of the infants, whereas acid concentration and peptic volume activity show a relatively steeper increase.


[Possibility of using higher plants in a life-support system on the moon]. The paper discusses the possibility of repeated termination of plant vegetation by prolonged darkness approximating the lunar night. This may be helpful for the incorporation of higher plants into the life support system of lunar bases, the solar light being used for illumination. In this connection vegetables (beet Bordeaux, turnip Petrovskaya, carrot Chantanet, dill, radish Virovsky white) and wheat (variety Sonora) were cultivated during the lunar light-dark cycle (i. e. 15 day light: 15 day dark). The experiments demonstrated that traditional plant products can be obtained under the conditions of lunar photoperiod. Grain of wheat grown during the lunar photoperiod were tested as seed material for further cultivation under similar conditions.


The effect of phospholipase D on the function of fragmented sarcoplasmic reticulum. Incubations of fresh preparations of fragmented sarcoplasmic reticulum (FSR) were carried out at pH 5.7. This pH was necessary for hydrolysis of phospholipids by phospholipase D. The pH did not influence calcium uptake or the activity of calcium-stimulated ATPase of FSR. Treatment of FSR with phospholipase D caused hydrolysis of the membrane phospholipids. The phosphatidic acid produced remained bound to the membrane. Increasing phospholipid cleavage was paralleled by loss of calcium uptake, which was complete when about two-thirds of the membrane phospholipids were hydrolyzed.


Multiple forms of acid phosphatase activity in Gaucher's disease. Although the primary genetic defect in all individuals with Gaucher's disease is a deficiency in glucocerebrosidase activity, the finding of marked elevations in splenic and serum acid phosphatase activity is almost as consistent a finding. Gaucher spleen and serum contain at least two forms of acid phosphatase that can be readily separated by chromatography on columns containing the cation exchange resin Sulphopropyl Sephadex. The major species of acid phosphatase (designated SP-I) contained in Triton X-100 (1% v/v) extracts of Gaucher spleen accounts for 65%--95% of the total activity and has the following properties: (1) it does not bind to the cation exchange column; (2) it exhibitis a pH optimum of 4.5--5.0; (3) it is inhibited by sodium fluoride (15 mM), L(+)-tartaric acid (20 mM), and beta-mercaptoethanol (2.1 M), and (4) it is resistant to inhibition by sodium dithionite (10 mM). The minor acid phosphatase activity (designated SP-II) present in extracts of Gaucher spleen has properties similar to those of the major species of acid phosphatase activity contained in serum from patients with Gaucher's disease: (1) it binds firmly to cation exchange columns (eluted by 0.5 M sodium chloride); (2) it exhibits a pH optimum of 5.0--6.0; (3) it is inhibited by sodium fluoride and sodium dithionite; and (4) it is resistant to inhibition by beta-mercaptoethanol (2.1 M) and L(+)-tartaric acid (20 mM). In addition, a second form of acid phosphatase that is tartrate resistant was found to be elevated in Gaucher serum. This form of serum acid phosphatase did not bind to Sulphopropyl Sephadex, was found to be significantly resistant to beta-mercaptoethanol (2.1 M), and was only partially inhibited by sodium dithionite (10 mM). The findings reported here indicate that at least three distinct forms of acid phosphatase activity are elevated in Gaucher's disease. Furthermore, the minor acid phosphatase activity contained in spleen homogenates has properties very similar to those of the major acid phosphatase activity observed to be present in serum of patients with Gaucher's disease. These data indicate that simple spleen spillage cannot account for the increased levels of serum acid phosphatase in patients with Gaucher's disease.


[Thermostability of glutamine synthetase in Candida tropicalis feed yeasts in Mg-, Mn- and Co-activated systems]. The thermostability of glutamine synthetase of Candida tropicalis was studied in systems activated by Mg, Mn and Co. The enzyme was found to be very thermolabile. Its thermostability depended on the nature of a bivalent cation used in the reaction mixture. Metals stabilized the enzyme since preincubation with metal cations increased the thermostability of glutamine synthetase. Purification of the enzyme preparation increased the stabilizing action of bivalent cations (Mg2+, Mn2+, Co2+).


[Effect of the composition of the medium and the conditions of Aspergillus foetidus cultivation on the biosynthesis of glucoamylase]. The optimal conditions for biosynthesis of exocellular glucoamylase were found in the course of submerged cultivation of Aspergillus foetidus ATCC 14916: pH 4.5 at the beginning of cultivation, cultivation for four days, a temperature of 30 degrees C, and an aeration in the fermenter of 1.5 volumes of the air per 1 volume of the medium with a stirring of 280 rpm. The material can be inoculated either as spores (1 X 10(7) spores per 100 ml of the medium) or a material germinated from spores (5% by volume). The composition of the medium was improved by using waste products of the agricultural industry (wheat and rice bran, yellow maize, etc.). The maximum biosynthesis of glucoamylase under the optimum conditions of cultivation was observed on crushed yellow maize (10--15%): up to 6.8--8.0 units of GIA (g of glucose per hour) per 1 ml of the cultural filtrate which was 3.4--4.0 times higher than on the original medium and 5--10 times higher than for Endomycopsis bispora, Endomycopsis sp. 20--9, Aspergillus niger and other cultures producing glucoamylase.


[Cellulase formation by soil yeasts of the genus Trichosporon and by microscopic fungi]. The production of celulases by a soil yeast belonging to the genus Trichosporon and by microscopic fungi of various systematic groups was studied. The yeast synthesized relatively small amounts of cellulases in contrast to microscopic fungi. The active culture producing cellulases was the fungus Trichoderma longibrachiatum 7-26.


[Cultivation of feed yeasts on media with carbohydrates]. The different culture of yeasts oxidizing hydrocarbons and the accompanying microflora are, under certain physico-chemical conditions, in metabiotic relationships which provide stability of the system. These relationships were studied with yeast cultures belonging to the Candida genus and oxidizing hydrocarbons. The relationships between the hydrocarbon oxidizing yeast Candida guilliermondii and other species of the genus Candida were determined by a combination of active and passive antogonism, as the result of which strains of this species completely dominated in the cenosis of fermenters.


[Origin of adifferentiated variants of Streptomyces roseoflavus in submerged culture]. Adifferentiated Nocardia-like variants (Nv) of Streptomyces roseoflavus var. roseofungini, similar in their cultural characteristics with the variant 1-68 described elsewhere (Nikitina, 1968; Kalakoutskii a. Nikitina, 1976), can be obtained not only in the course of growth of the parent culture on a solid medium containing fructose in the form of secondary colonies, but also during submerged cultivation with stirring. Interruptions in aeration do not affect accumulation of the cells of such variants in the medium. A direct correlation has been established between the amount of biomass and the number of Nv colonies when submerged mycelium is inoculated into a solid medium.


[Effectiveness of strains of clover nodule bacteria adapted to the acid reaction of the soil]. The effectiveness and virulence of clover nodule bacterial strains of different origin was studied as they were adapted to acid pH (4.8) of soil in the course of seven years. In spite of their ecological adaptation (strains from acid and neutral soils), the majority of strains (10 among 20) grew well enough upon numerous transfers onto acid media under new conditions. However, only some adapted strains were more effective, than non-adapted ones on acid soil Storage of the strains on acid media for a prolonged time had a positive effect on their virulence, when clover was inoculated on calcareous soil.


Isoenzymes of human liver alpha-L-fucosidase: chemical relationship, kinetic studies, and immunochemical characterization. The chemical relationship of the seven forms of human liver alpha-L fucosidase has been studied by isoelectric focusing of neuraminidase- and sialytransferase-treated preparations of alpha-L-fucosidase. Neuraminidase treatment leads to a decrease in the activity of the more acidic forms (IV-VII) and a concimitant increase in the activity of the more neutral forms (I-II). Incubation of the neuraminidase-treated fucosidase (forms I-III) with radiolabelled cytidine monophosphate-N-3H-acetylneuraminic acid and an enriched preparation of sialytransferase devoid of fucosidase activity led to regeneration of the more acidic fucosidase isoenzymes (IV-VII) with the same isoelectric points and in nearly the same proportion as before neuraminidase treatment. These experiments suggest that the isoenzymes of human liver alpha-L-fucosidase are related, at least in part, by sialic acid residues. The seven isoenzymes of purified human liver alpha-L-fucosidase have been separated by preparative isoelectric focusing and characterized kinetically and immunochemically. Differences in Michaelis constants (Km's) and pH optimum curves were found for some of the isoenzymes. All seven isoenzymes were immunoprecipitated using the IgG fraction of anti-alpha-L-fucosidase antiserum suggesting that the presence of sialic acid residues does not affect the antigenicity of the forms of alpha-L-fucosidase.


[On the pharmacological treatment of acute spasmodic laryngitis (Pseudo-Croup) (author's transl)]. Three aspects of the pharmacological treatment of pseudo-croup are discussed in this article. Sedatives should be given only when close clinical supervision is guaranteed. Otherwise the apparent drowsiness observed in patients passing from stage III to stage IV of the disease may be masked or misinterpreted due to these sedatives. Emetics used occasionally in the USA, and also frequently administered in Europe to the turn of the present century, are no longer used for treatment in Germany. There are no controlled studies showing their effectiveness. In contrast to the common practice in Germany, glucocorticosteroids are less frequently used in the USA. Theoretical considerations suggest their usefulness, but to data there is still no convincing proof of their clinical effectiveness.


[Atrial tachycardia with atrio-ventricular block in infancy and childhood (author's transl)]. Atrial tachycardia with atrio-ventricular block is a very rare arrhythmia in infancy and childhood. We saw ten patients between 3 days and fifteen years old. Six of them had a congenital heart disease, in four cases we could'nt find any cardiac or extracardiac illness. In every case digitalis intoxication was excluded. Clinical findings, electrocardiographic features, therapy and prognosis are reported.


[Electrical conductivity of isoionic DNA solutions]. Conductance of the DNA isoionic solutions obtained from aqueous solutions of its sodium salt using a column with mixed bed ionexchangers has been investigated. It has been found that in Kohlrausch's plots equivalent conductance slightly decreased with polymer concentration. Comparison of the degree of ionization (alpha) calculated from conductance measurements with those calculated from experimental pH values of isoionic solutions at different polymer concentrations revealed that the mobility of dissociating counterations decreases significantly under the influence of the electrostatic potential of the macroion.


[Treatment of hypertension with a combination of pyridopyridazine derivative with a beta-receptor blocker. Studies on the hemodynamic course (author's transl)]. 22 pateints with a fixed, sometimes therapy-resistant (N = 18) hypertension were treated with a new antihypertensive agent, the pyridopyridazine derivative BQ 22-708 and hemodynamically investigated. 14 hypertensives received single oral doses of between 5 and 15 mg, 8 other patients also received additional treatment with a beta-receptor blocker (0.8 mg pindolol i.v.) or calcium antagonist (10 mg verapamil i.v.). In hypertension which is only poorly controllable or resistant, therapy with BQ 22-708 combined with beta-receptor offers a genuine alternative medication.


[Cardiac neurosis in everyday general medical practice (author's transl)]. From results of the study of 37 patients with cardiac neurosis, new conceptual relationships to explain this condition are developed. Cardiac neurosis is explained as a disturbance of the "psycho-neurovegetative self-regulation mechanism" (servomechanisms). The authors show how such a conception, which has been borrowed from the servomechanism technique, may favourably influence the diagnosis and therapy of this condition. It has been tried in that way to link the two opposites: organic and functional causes. Some therapeutic approaches adapted to the structure of everyday general medical practice are suggested.


Variable content of histaminase, L-dopa decarboxylase and calcitonin in small-cell carcinoma of the lung. Biologic and clinical implications. To ascertain whether the content of endocrine markers is constant in small-cell carcinoma of the lung, levels of three markers of medullary thyroid carcinoma were studied in this tumor. Histaminase was increased in six of six primary tumors (three to 14,000 times), L-dopa decarboxylase in four of six (six to 30 times), and calcitonin in one of one (eight times) over levels in adjacent lung. Marker levels in mediastinal metastases reflected those in primary tumors in four of five patients. However, in four of seven, multiple hepatic metastases contained low to absent levels despite simultaneously high values in chest lesions. Immunohistochemical studies of histaminase revealed that within each primary tumor different cells contained different amounts of the enzyme. Since marker content varied between tumor cells, between primary tumors and between metastases in individual patients we conclude that circulating levels of these three markers cannot be expected necessarily to mirror tumor burden in patients with small-cell lung tumors.


Current problems in the control of mosquitoes. Since the incidence of mosquito-borne disease was drastically reduced by the use of DDT in the 1950s, the problem has again worsened and no single method of eradication seems likely to achieve lasting success. It now seems that the best hope lies in the combination of various methods of chemical control and perhaps genetic control to rid the Third World of its most important disease sector.


[Value of brain metabolic studies in post-traumatic coma]. In 34 comatose patients in the acute phase, the mean hemispheric CBF is lowered as well as the CMRO2, with a quite good relation between these values and the coma level and prognostic so. The cerebral response to a PaCO2 range indicates a quite good relation with the coma level (the lowest value in the most severe comas). The cerebral autoregulation study, using Aramine induced hypertension, can separate the cases with a present autoregulation and the cases with a loss of autoregulation (the most severe and the poorest prognosis). In dynamic conditions (variation of the PaCO2 or Aramine induced hypertension), the change in CMRO2 is interesting : rather good prognosis among the patients with a normal metabolic autoregulation - poor prognosis among those whose metabolic autoregulation is lost.


[Alkaline urine in the aged].

[Alkaline urine in the aged]. Considering the frequent finding of alkalinity in the urines of elderly people of both sexes, we can propose many hypothesis among which the most probable seems a latent renal failure with secondary hyperparathyroidism and consequent alkalinization of the urines. We can presume that such a frequent alkalinity of the urines in old subjects has an important role in the frequent infectious of the urinary tract of elderly people.


[Hemodynamic effects of antihypertensive drugs]. The physiopathology of hypertension is not fully understood, though the haemodynamic pictures accompanying the various arterial forms have been established. Clinical employment of antihypertensive drugs should tend to correct these pictures and the extent to which this is true is examined in the light of the latest evidence with respect to the drugs most commonly used.


Urinary sulfate excretion in children with classic renal tubular acidosis. These studies quantitat the significantly elevated sulfate excretion in the urine of children with classic renal tubular acidosis as compared to six weight-matched controls (1.4 +/-0.5 vs. 0.7 +/- 0.2 mEq/kg/day;p less than 0.05). This sulfate loss may result in a subclinical sulfate deficiency which may contribute to a chondroitin sulfate metabolic disorder.,and thus contribute to the growth failure in children with real tubular acidosis. Futhermore, elevated urinary sulfate excretion may be an early diagnostic finding in infancy prior to the manifestation of renal acidification defects. It is not known whether the sulfate loss is related to a primary renal tubular defect or secondary to metabolic acidosis, although the increased sulfate excretion persisting after correction of metabolic acidosis would tend to suggest a primary defect. However, the normal plasma sulfate concentration and the relatively short period of study permit no definitive answer at this time.


Hydrogen ion production secondary to metabolism of sulfur-amino acids and organic acids. The rate of acid production in infants and young children was quoted to be 1-2 mEq/kg/day but in fact, the sole reference used in all these instances did not permit such quantitation. In order to evaluate the rate of endogenous acid production, we applied the quantitative technique of Lennon et al. in the following normal subjects, where column A represents hydrogen ion released from metabolism of sulfur-containing amion acids; column B represents hydrogen ion released from incomplete oxidation of organic acids; the sum A and B gives an estimation of endogenous net acid production (NAP).


The effect of atropine on the lower esophageal sphincter in late pregnancy. Intraluminal gastroesophageal pressure and pH studies have been performed on 8 nonpregnant women, 10 pregnant women with heartburn, and 10 pregnant women without heartburn. Each patient was tested under resting conditions and after intravenous injection of 0.6 mg atropine. In both groups of pregnant patients the intragastric pressure was found to be higher than that of the nonpregnant subjects. The stomach to lower esophageal sphincter pressure (LESP) gradient under resting conditions was least in the pregnant patients with heartburn. After the administration of atropine, a fall in the LESP occurred in all 3 groups of patients which was most profound in the nonpregnant subjects and in the pregnant patients without heartburn. These changes and the pH recordings of the lower esophagus indicate the adverse effect that atropine has on the competency of the LESP both in pregnancy and in the nonpregnant state. Atropine should therefore be used with caution as a premedicant and preferably combined with metoclopramide (Maxolon).


Deceleration/contraction ratios as an index of fetal health during labor. Internal FHR tracings of 259 patients with variable decelerations and 49 with late decelerations were analyzed for frequency and severity of the pattern in an attempt to correlate with fetal pH or Apgar score at 1 minute. A significant increase in the risk of fetal acidosis was associated with a frequency of greater than 30% variable decelerations and 20% late decelerations. It also increased with the increase of severity of the pattern. Variable decelerations are not good predictors of Apgar scores less than or equal to 6 at 1 minute, but late decelerations are predictive when they occur more than 20% of the time. The fetus with variable decelerations a normal pH can be observed through the first stage of labor with a high degree of confidence. However, when a 20% or more moderate-to-severe late deceleration/contraction ratio is present, intervention is necessary even if scalp blood pH is normal. The FHR deceleration/uterine contraction (UC) ratio is a simple bedside technique for assessing fetal outcome.


gamma-Glutamyl transpeptidase activity in sheep serum: normal values and an evaluation of its potential for detecting liver involvement in experimental lupinosis. A brief survey of the literature on gamma-glutamyl transpeptidase (gamma-GT) activity is included in this study. The levels of activity in the serum of normal Merino sheep (13,6-32,4 mI.U/ml) were ascertained as a preliminary to following the activity through the entire course of experimentally induced ovine lupinosis, a hepatotoxicosis caused by Phomopsis leptostromiformis (Kühn) Bubák. The response of the serum level of gamma-GT activity to the course of the disease was compared with that of glutamate oxaloacetic transaminase (GOT) and 2 liver function tests for the purpose of assessing its potential application in the study of this mycotoxicosis. Because the levels of activity of gamma-GT were more valuable for the early diagnosis of low grade acute intoxication and the detection of chronic liver involvement while those of GOT gave better information on the development of severe acute hepato-cellular damage, these 2 enzymes, considered together, were found to give the best information on the course of the toxicosis. The changes in gamma-GT activity during various stages of intoxication were also related to the histopathological lesions in the liver.


The metabolic profile of adult Fasciola hepatica obtained from rafoxanide-treated sheep. Sheep infected with adult Fasciola hepatica were drenched with rafoxanide. At 4, 8, 16 and 24 h after drenching the sheep were killed and the flukes removed, washed and rapidly frozen in liquid nitrogen. The content of key metabolites in the fermentation pathway were determined and compared with those in control F. hepatica, whose hosts were not treated with rafoxanide. Rafoxanide decreased glycogen, malate, NADH and ATP levels. The level of other metabolites in the pathway increased for the first 8-16 h after rafoxanide treatment. The marked decrease in ATP and glycogen, and the increase in total [NAD+]/[NADH] and [oxaloacetate]/[malate], together with the changed content of other metabolites, led to the conclusion that the mode of action of rafoxanide against F. hepatica in vivo is by uncoupling oxidative phosphorylation.


A basis for migraine therapy- the autonomic theory reappraised. The concept that migraine results from an initial vasoconstriction due to increased release of noradrenaline from the sympathetic nerves to cranial blood vessels has been reappraised in the light of recently acquired knowledge of the mechanisms of action of drugs used in the treatment of migraine, physiological and pharmacological evidence implicating noradrenaline, and the observations by others that several migraine variants may be associated with some degree of sympathetic overactivity. If the theory is correct, it suggests that both prophylaxis and management of the acute condition should be possible by means of selective alpha-adrenoceptor antagonism. The use of drugs with potentially dangerous vasoconstrictor properties appears to be unnecessary. The suggestion is made that the increased adrenergic activity might result from changes within the hypothalamus.


[Changes in the urinary excretion of gamma-glutamyltranspeptidase, leucine aminopeptidas and alkaline phosphatase in the combined action of ethylene glycol and high temperature]. Gamma-glutamyl transpeptidase (gamma--GTP), leucine aminopeptidase (LAP) and alkaline phosphatase (AP) excretion in rats is followed in dynamics (2, 8, 15, 30 and 90 days) upon isolated and combined treatment with ethylene glycol (EG) at dose 1/8 LD50 and temperature of the environment 35 degrees C. Under the effect of high temperature an increase in the excretion of enzymes in the early observation terms is noted. The independent application of the noxa causes a reduction in gamma--GTR and LAP excretion, and an increase in AP. The temperature factor attenuates the toxic effect of EG relative to the enzymes under study at the end of the observation period. Changes in gamma--GTP excretion are considered as the earliest and most sensitive sign of tubular lesions.


Mode of action of colicin Ia: effect of colicin on the Escherichia coli proton electrochemical gradient. By use of the technique of flow dialysis, the membrane potential (deltapsi) and pH gradient (deltapH) have been measured in colicin Ia-treated Escherichia coli K-12 cells and in membrane vesicles prepared from such cells. Although such cells and vesicles are able to generate a transmembrane deltapH at pH 5.5, they do not generate a transmembrane deltapsi. Glucose-6-phospate uptake by cells is shown to be stimulated at pH 5.5 and inhibited at pH 7.5 by colicin Ia treatment. On the other hand, proline uptake is demonstrated to be inhibited progressively at pH 5.5, 6.6, and 7.5 in colicin Ia-treated cells. These data provide strong evidence for a colicin Ia-induced membrane deplorization and indicate that the membrane becomes permeable to ion(s) other than protons after treatment with colicin Ia.


Proposed temperature-dependent conformational transition in D-amino acid oxidase: a differential scanning microcalorimetric study. A number of authors have reported observations on D-amino acid oxidase [D-amino acid: O2 oxidoreductase (deaminating), EC 1.4.3.3.] that they have interpreted in terms of a temperature-dependent conformational transition having a van't Hoff enthalpy amounting to more than 1 cal per g of protein (1 cal = 4.184J). No indication of this transition is obtained by using a differential scanning calorimeter having a sensitivity considerably in excess of that required to detect such a transition. The implications of this discrepancy are discussed.


Elimination of cooperativity in aspartate transcarbamylase by nitration of a single tyrosine residue. In a previous report [Landfear, S. M., Lipscomb, W. N. & Evans, D.R. (1978) J. Biol. Chem. 253, 3988--3996] we demonstrated that tetranitromethane can be employed to nitrate a limited number of tyrosine residues in aspartate transcarbamylase (carbamoylphosphate:L-aspartate carbamoyltransferase, EC 2.1.3.2); such modification eliminates cooperativity, feedback inhibition, and enzymatic activity, and reduces binding of the feedback inhibitor cytidine triphosphate. Cooperativity is lost more rapidly than other properties, and this loss correlates with the nitration of a single tyrosine residue. In this paper, we describe the saturation kinetics of hybrid species constructed from nitrated subunits of one type (either catalytic or regulatory) and native subunits of the other type. We conclude that the modification responsible for loss of cooperativity is on the catalytic subunit. The tryptic peptide containing this modification has been isolated and identified.


Heat-stable enterotoxin of Escherichia coli: in vitro effects on guanylate cyclase activity, cyclic GMP concentration, and ion transport in small intestine. A partially purified preparation of the heat-stable enterotoxin of Escherichia coli caused a rapid and persistent increase in electric potential difference and short-circuit current when added in vitro to the luminal surface of isolated rabbit ileal mucosa. As little as 1 ng/ml produced an easily detectable response. Under short-circuit condition, the enterotoxin abolished net Cl- absorption; this change was half that produced by theophylline, which stimulated net secretion. The enterotoxin did not change cyclic AMP concentration but caused large and persistent increases in cyclic GMP concentration. The electrical and nucleotide responses exhibited similar and unusually broad concentration-dependences and maximal effects could not be demonstrated. Theophylline elevated cyclic GMP concentration 3-fold both in the presence and absense of the enterotoxin, suggesting no effect of the toxin on cyclic GMP phosphodiesterase. Guanylate cyclase [GTP pyrophosphatelyase(cyclizing); EC 4.6.1.2] activity in a crude membrane fraction from intestinal epithelial cells was stimulated 7-fold by the enterotoxin. These results suggest that guanylate cyclase stimulation is the basis for the toxin's diarrheagenic effect.


Hemopoietic stem cell transplantation using mouse bone marrow and spleen cells fractionated by lectins. Mouse bone marrow and spleen cells were fractionated with the aid of soybean agglutinin and peanut agglutinin. A test for spleen colony-forming units in the isolated fractions showed that the hemopoietic stem cells are agglutinated by both of these lectins. The capacity of the agglutinated fractions to reconstitute lethally irradiated allogeneic mice was investigated. A sequential fractionation of splenocytes from SWR donors by soybean agglutinin and peanut agglutinin, or a single fractionation by soybean agglutinin of splenocytes from BALB/c donors, afforded a cell fraction that successfully reconstituted lethally irradiated (BALB/c X C57BL/6)F1 mice, without complications due to graft-versus-host reaction.


Beneficial and detrimental actions of histamine H1- and H2-receptor antagonists in circulatory shock. This study explores the use of both histamine H(1)- and H(2)-receptor antagonists in two different forms of circulatory shock and suggests that histamine may be involved in more than one way in the pathophysiology of circulatory shock. Various single doses of diphenhydramine, chlorpheniramine, promethazine, and burimamide were administered intravenously to Wistar rats subjected to hemorrhagic or bowel ischemia shock. Cumulative survival and mortality, as well as arterial blood pressures and microhematocrits, were monitored. Pretreatment of the animals with the three different H(1)-receptor antagonists exerted significant protection against both forms of shock. Rats pretreated with the H(2)-receptor antagonist, burimamide, demonstrated an exacerbated mortality after induction of shock. Animals pretreated with H(1)-receptor antagonists showed significantly higher mean arterial blood pressure, greater compensatory rebound of blood pressure after induction of shock, and greater responses to transfusion after hemorrhage than control, shocked animals. Similarly, rats pretreated with the H(1)-receptor blockers demonstrated significantly greater compensatory hemodilution which continued late in shock. In marked contrast, rats pretreated with burimamide exhibited opposite effects after hemorrhage and bowel ischemia, i.e., significant falls in blood pressure, lack of compensatory rebound and response to transfusion of shed blood, and a progressive hemoconcentration. This report clearly demonstrates beneficial actions of histamine H(1)-receptor antagonists and detrimental effects of H(2)-receptor antagonists on survival and other parameters in these forms of circulatory shock.