Studies on acid lipase, and E 600-resistant acid esterase activities in human tissue homogenates. Detailed comparison of acid lipase and acid esterase activities of human spleen, liver and kidney homogenates has been carried out by means of the following substrates: 14C-tripalmitin, alpha-naphthyl acetate, alpha-naphthyl butyrate, alpha-naphthyl laurate, p-nitro-phenyl acetate, butyrate and laurate. In addition, homogenates of the three tissues were subjected to isoelectric focusing in polyacrylamide gels and histochemical staining with the above mentioned naphthyl substrates in the presence and absence of the organophosphate esterase inhibitor diethyl-p-nitrophenyl phosphate (E 600). These studies provide extensive support for the proposal that E 600-resistant acid naphthyl butyryl and lauryl esterase activities in human tissues derive largely from the enzyme acid lipase. The studies suggest that the most specific chromogenic substrate for this enzyme at a biochemical and histochemical level is alpha-napthyl laurate in the presence of E600 (3 X 10(-6) M).


Multiple forms with glucose 6-phosphate dehydrogenase activity in Musca domestica L. as revealed by electrophoresis on cellulose acetate gel. Single newly emerged males of Musca domestica, WHO strain, usually show five electrophoretic bands of glucose 6-phosphate dehydrogenase (G6PD) activity. Of these five molecular forms, designated with Roman numerals in order from the origin, we have considered the first three: these have been characterized with respect to their substrate and coenzyme specificity and to their sensitivity to some sulfhydryl inhibitors. The data show band III to be G6P specific, nicotinamide adenine dinucleotide phosphate dependent and to be a type I enzyme according to Kamada and Hori's classification. Bands I and II, on the other hand, show wide substrate specificity and low sensitivity to the sulfhydryl inhibitors assayed. In addition, in the absence of an exogenous substrate and in the presence of nicotinamide adenine dinucleotide as a coenzyme, fairly weak bands, which can be ascribed to the so called "nothing dehydrogenase" effect, are seen in the position I and II. Nevertheless, the data reported do not allow a clear definition of the enzymatic type corresponding to bands I and II of G6PD activity.


The in vitro conversion of a specific molecular form of glucose 6-phosphate dehydrogenase from Musca domestica L. A nicotinamide adenine dinucleotide phosphate dependent glucose 6-phosphate dehydrogenase (G6PD), belonging to type I of Kamada and Hori's classification, is present on the zymograms of newly emerged males of Musca domestica. It is capable of undergoing tryptic degradation and being thus transformed into a different active enzymatic form, with some of its catalytic properties unchanged, but with different electrophoretic mobility. We show in this paper that this specific G6PD form of gut origin in M. domestica is not a tissue-specific enzyme, but rather a product of hydrolytic degradation by gut proteinases which act during the process of homogenization. Besides, the G6PD of type I in the housefly is shown to be sensitive to the "storage effect" and to protection by mercaptoethanol, contrary to its hydrolytic gut form which is not sensitive to these processes. In this connection, we discuss the possible reasons for these differences in behavior.


Syngeneic GvH induced in popliteal lymph nodes by spleen cells of old C57BL/6 mice. The frequency of autoimmune processes seems to increase with age. We have studied here whether lymphoid cells of aged mice have the potential to express autoreactivity by the use of the in vivo graft-vs-host (GvH) assay. It was found that spleen cells from old (104 weeks) C57BL mice caused significant enlargement of the popliteal lymph node upon injection into the footpads of syngeneic young or old recipients. Histologically this enlargement presented characteristics of a GvH reaction. This effect, which was not abolished by irradiation of the hosts, was totally cancelled by in vitro irradiation or by anti-theta treatment of the donor cells. These results indicate that T cells from aged mice have the potential to manifest autoimmune reactivity.


Effect of prolonged occlusion on the microbial flora, pH, carbon dioxide and transepidermal water loss on human skin. The effects of prolonged occlusion on the normal microbial skin flora, pH, transepidermal water loss (TEWL) and carbon dioxide emission rate (CDER) were studied. The total average counts before occlusion were 1.8 X 10(2)/cm(2) and increased to 4.5 X 10(6) on day 5. The highest counts were noted on day 4(9.8 X 10(7)/cm(2)). The composition changed: controls comprised of 63% coagulase negative staphylococci, 6% micrococci, 17% diphtheroids and 6% bacilli. After 5 days of occlusion, the percent compositon was: 63% coagulase negative staphylococci, 11% micrococci, 4% diphtheroids, 19% lipophilic diphtheroids and less than 0.003% gram negative rods. The pH of the skin before occlusion was 4.38 and increased to 7.05 on day 5. After 5 days of occlusion TEWL increased from 0.56 mg/cm2/hr to 1.87 mg/cm2/hr and CO2 emmission increased from 25 nl/cm2/min to 118 nl/cm2/min.


Models for assessing the effect of toxicants on immunocompetence in mice. Part I: The effect of diphtheria, pertussis, and tetanus vaccine on antibody response to type III pneumococcal polysaccharide. This study characterizes the response of BALB/c mice to Type III pneumococcal polysaccharides (S3) administered alone or with a vaccine consisting of diphtheria toxoid, pertussis vaccine, and tetanus toxoid (DPT). The usefulness of this vaccine in toxicological protocols for determining immunocompetence is discussed. There were significant differences in antibody titers depending upon the route of injection and whether the S3 was administered in saline or mixed with DPT. Subcutaneous injections induced higher antibody titers to S3 than did intraperitoneal injections. The effect of DPT varied depending upon the manner of administration. Subcutaneous injections of DPT mixed with S3 increased the S3 antibody titers to a dose of 0.1 microgram S3, but had little or no effect at 0.25, 0.5, and 0.75 microgram. Antibody titers were suppressed only at the highest dose administered (1.0 microgram). On the other hand, when administered intraperitoneally DPT has a consistently suppressive effect on 5 day peak S3 antibody titers. Intraperitoneal injections of DPT appeared to delay the rate of antibody induction, since antibody titers at 14 and 21 days were higher in the DPT treated mice than in those animals receiving S3 alone.


Enteroendocrine APUD cells in the digestive tract of larval Barbus conchonius (Teleostei, Cyprinidae). The development of Barbus conchonius is described with special attention to the differentiation of the gut. Amine precursor uptake and decarboxylation (APUD) are present in enteroendocrine cells during development, whereas these processes are lacking in adult specimens. The first APUD cells originate on the fourth day of development in the anterior part of the gut and on the fifth day in the caudal areas. The APUD facility of the cells disappears within 2 days, and after the 6th day APUD cells can no longer be distinguished in the intestinal epithelium. The first APUD cells were obsserved when four types of enteroendocrine cells were recognized with the electron microscope. These enteroendocrine cells contain granules of different electron densities, and microtubules and cilia can be observed. Some enteroendocrine-like cells are found below the basement membrane of the intestinal epithelium, indicating a possible extra-endodermal origin. APUD cells, except melanoblasts, have not been found migrating from the neural crest in ventral direction. The origin of the enteroendocrine cells of B. conchonius is discussed.


The mouse gut T lymphocyte, a novel type of T cell. Nature, origin, and traffic in mice in normal and graft-versus-host conditions. Lymphocytes of the mouse intestinal mucosa, identified in tissue sections or purified suspensions of intraepithelial lymphocytes as T cells (gut T lymphocytes [GTL]), were studied in normal mice or in beige mice (the equivalent of the Chediak-Higashi syndrome in man, characterized by giant granules in various cell types, including mast cells). Mice were studied in normal or in germ-free conditions, or during a graft versus host (GVH) reaction resulting from the injection of parental thymocytes into lethally irradiated F1 mice, a condition leading to massive accumulation of T lymphocytes of donor origin in the host gut mucosa. In normal as well as in GVH conditions, a high percentage of the gut IE lymphocytes contain granules (up to 80% in the beige mouse). These granules have ultrastructural, hostochemical and other features resembling those of mast cell granules; in beige mice, up to 50% of them can be shown to contain histamine. Granulated T cells are also found in the lamina propria. It appears that the GTL may progressively lose their surface T antigens when the granules become more developed. Kinetics of [3H]TdR labeling of the GTL, transfer experiments with T cells of various origins, selective [3H]TdR labeling and selective irradiation of the Peyer's patches (PP), and effect of thoraic duct (TD) drainage led to the conclusion that GTL are the progeny of T cells stimulated to divide in the PP microenvironment, which endows them with a gut-homing tendency. From the PP, these cells follow a cycle, migrating to the TD and to the blood to colonize the whole intestinal mucosa, the majority of them as dividing cells undergoing a single round of traffic, with some probably able to recirculate and becoming a more long-lived variety. Antigenic stimulation within the PP is necessary for the emergence of GTL progenitors, but their gut-homing property is unrelated to the antigen as shown with fetal gut grafts, notably in GVH where grafts syngeneic to the host or donor become similarly infiltrated by GTL. On the basis of their properties and of further evidence to be reported elsewhere, it is proposed that GTL belong to a special class of T lymphocytes, related to the immune defenses of the mucosal systems in general, and capable of acting as progenitors of mucosal mast cells.


Facilitation of transmitter release at squid synapses. Facilitation is shown to decay as a compound exponential with two time constants (T1, T2) at both giant and non-giant synapses in squid stellate ganglia bathed in solutions having low extracellular calcium concentrations ([Ca++]o). Maximum values of facilitation (F1) were significantly larger, and T1 was significantly smaller in giant than non-giant synapses.  Decreases in [Ca++]o or increases in [Mn++]o had variable effects on T1 and F1, whereas decreases in temperature increased T1 but had insignificant effects on F1. The growth of facilitation during short trains of equal interval stimuli was adequately predicted by the linear summation model developed by Mallart and Martin (1967. J. Physiol. (Lond.). 193:676--694) for frog neuromuscular junctions. This result suggests that the underlying mechanisms of facilitation are similar in squid and other synapses which release many transmitter quanta.


Presynaptic potentials and facilitation of transmitter release in the squid giant synapse. Presynaptic potentials were studied during facilitation of transmitter release in the squid giant synapse. Changes in action potentials were found to cause some, but not all, of the facilitation during twin-pulse stimulation. During trains of action potentials, there were no progressive changes in presynaptic action potentials which could account for the growth of facilitation. Facilitation could still be detected in terminals which had undergone conditioning depolarization or hyperpolarization. Facilitation could be produced by small action potentials in low [Ca++]o and by small depolarizations in the presence of tetrodotoxin. Although the production of facilitation varied somewhat with presynaptic depolarization, nevertheless, approximately equal amounts of facilitation could be produced by depolarizations which caused the release of very different amounts of transmitter.


Some characteristics of Ureaplasma urealyticum. Urease activity in a simple buffer: effect of metal ions and sulphydryl inhibitors. Urealytic activity of the cytoplasmic fraction of Ureaplasma urealyticum prepared by digitonin lysis was assayed in a simple buffer system (HEPES plus EDTA) by measuring the release of 14CO2 from [14C]urea. The Km of this preparation agreed with our previous observations of the same activity measured in a more complex reaction mixture. The substrate concentration at which maximum velocity occurred was approximately 20 mM. The activity was sensitive to heavy metals and inhibitors which react with sulphydryl groups such as N-ethylmaleimide and p-chloromercuribenzoate. It was not inhibited by Ca2+ or Mg2+ or by the reaction products, ammonia and carbon dioxide.


Photosensitization of herpes simplex virus type 1 with neutral red. Commercial neutral red (NR) originally containing at least 8 components was purified by thin layer chromatography. Herpes simplex virus type 1 (HSV-1) treated in vitro with 30 microgram/ml of purified NR became sensitive to light inactivation within 2 min but rapidly lost this sensitivity upon dilution. Similarly, virus grown in the presence of NR lost its photosensitivity upon dilution of the virus stock. In both cases the kinetics of inactivation appeared to be multi-hit. Photoinactivation of intracellular virus was most effective when NR was applied between 6 and 12 h post-infection. The most efficient inactivation occurred when virus at pH 8.8 was irradiated by light at a wavelength of 470 nm.


Secondary depression in anxiety neurosis. Forty-four per cent of 112 patients with anxiety neurosis reported episodes of depression during the course of their illness compared with only 7 per cent of surgical controls. Although the majority developed in response to environmental circumstances and were of brief duration, they commonly led to psychiatric treatment or hospitalization in this group of patients. Patients who developed this complication were shown to have a more chronic and severe underlying illness.


Frightening dreams and dosage schedule of tricyclic and neuroleptic drugs. One previous study reported that psychiatric inpatients on bedtime-only doses of tricyclic or neuroleptic drugs reported more frequent frightening dreams than did those on divided daily doses. As a further test of this hypothesis, and in order to see whether differences in frequency of frightening dreams were a function of differences in total dream recall, we administered a questionnaire to outpatients in a Veterans Administration Hospital Mental Hygiene Clinic, asking about frequency of dream recall, frequency of frightening dream recall, and doses and times of any medications taken. Questionnaire reports of medications were checked with the medical record of the patient; for 48 patients on tricyclic or neuroleptic drugs the reports agreed, and the data of these patients were analyzed. Our findings corroborated the previous report of more frequent frightening dreams in patients on bedtime-only dosage schedules (p less than .01). In addition, we found no significant difference between the two groups with respect to frequency of dream recall; thus a difference in the affect of dreams, rather than a difference in quantity of dream recall, constituted the difference between the two groups. When a patient on bedtime doses of tricyclic or neuroleptic drugs has undesirable frightening dreams, the clinician should consider a change to divided daily doses.


Induced hypotension for orthognathic surgery. Achieving satisfactory hemostasis during orthognathic surgery may be difficult because of the extensive vascularity of facial structures. Hypotensive anesthetic techniques provide clear operative fields by altering regional tissue perfusion through the use of systemic vasodilators, ganglionic blocking agents, and positioning of the patient. Thorough monitoring during surgery, careful selection of patients, and close communication between the surgeon and anesthesiologist permit safe anesthesia, can decrease operating time, and usually obviate the need for transfusions.


Hemolymph of Anopheles stephensi from noninfected and Plasmodium berghei-infected mosquitoes. 1. Collection procedure and physical characteristics. Hemolymph was collected from adult female Anopheles stephensi by centrifugation of incised mosquitoes. Approximately 0.1 muliter was collected from each recently emerged mosquito, although smaller amounts were recovered with increasing age of the mosquito. Determinations were made of the pH, osmotic pressure, and specific gravity of this hemolymph at various times during the life of the adult mosquito. The values obtained were within the ranges found for other insects. Hemolymph collected from mosquitoes fed on hamsters infected with Plasmodium berghei had different values than hemolymph from mosquitoes fed on noninfected hamsters. This probably was due to differences between the quality of these 2 types of blood meals, rather than to the direct effects of the malaria parasite on the infected mosquito itself.


The effect of some formulation and process variables on the surface roughness of film-coated tablets. The effect of some formulation and process variables on the surface appearance of film-coated tablets has been examined by measuring the arithmetic mean roughness, Ra, values across the faces of tablets before and after they were coated with hydroxypropyl methyl-cellulose. For all tablet cores except those that were very porous, film coating resulted in an increasing surface roughness; for very porous cores a decrease was found. Tablets with rough surfaces were produced by coating with low molecular weight grades of the polymer; increasing the polymer molecular weight resulted in a smoother finish with a minimum roughness at intermediate molecular weight grades. Increasing the polymer concentration above 2% w/v caused an increase in roughness as did increasing film thickness to 140 micrometer. There was a minimum in roughness at film thickness of 20 micrometer. The addition of pigment in low concentrations (0--25% v/v) caused a marginal increase in surface roughness but at concentrations above the critical pigment volume concentration, the surfaces were very rough. The results illustrate the potential of the method in the optimization of film formulations and process conditions during product development.


Effects of temperature on the tensile strength of pharmaceutical powders. Measurements have been made of the tensile strengths of fatty acids, lactose and of an oxtetracycline and a paracetamol tablet formulation at temperatures between -20 and +90 degrees. The measurements were made on compressed tablets and on consolidated beds of powders, using a diametral compression tester and a split plate tensile tester which had been designed to operate over this range of temperature. The tensile strengths of all the materials increase with temperature and values are given for the activation energies of bonding between particles.


Lithium-6 stable isotope determination by atomic absorption spectroscopy and its application to pharmacokinetic studies in man. No useful radioisotope of lithium exists to assist in the study of its biochemical pharmacology. A simple method has been developed for the determination of the stable isotope 6Li by atomic absorption spectroscopy (AAS). The technique is applicable in any laboratory equipped with simple AAS apparatus. Analysis of total lithium content (E) by flame emission spectroscopy is followed by separate determination (A6 and A7) of atomic absorption by the sample of the light emitted by separate hollow cathode lamps manufactured from the isotopes 6Li and 7Li. The standard curve of absorption ratio (A6/A7) against isotopic ratio (6Li/7Li) at any concentration (E) is an exponential which may be solved using a simple programmable calculator. Application of this method to the study of the pharmacokinetics of 6Li administered to 4 normal volunteers previously loaded with 7Li suggests that the rate of appearance of lithium in blood is unaffected by the previous state of lithium loading.


Sustained release of sulphadiazine.

Sustained release of sulphadiazine. An implantable system was developed which released sulphadiazine in mice over an extended period of time efficacious against infective challenges by Plasmodium berghei. The most successful preparation was a copolymer of L(+)-lactic acid + (+/-)-lactic acid (90 and 10% by weight, respectively) with a molecular weight of 150 000, with which sulphadiazine was mixed at 33.3% of the total weight, in a formulation as beads of 1.5 mm diameter. This preparation released sulphadiazine at a nearly constant rate over three months as measured by the appearance in urine of mice of radioactivity from [35S] sulphadiazine in transplanted material. When implanted in mice, the beads gave effective protection against repetitive (weekly) infective challenges with P. berghei by implanted beads at dosages equivalent to 57 mg kg(-1) sulphadiazine and greater over 21 weeks.


Distribution of pellets in the gastrointestinal tract. The influence on transit time exerted by the density or diameter of pellets. The influence on transit time exerted by the density or diameter of pellets has been established in six ileostomy subjects. An increase in density from 1.0 to 1.6 significantly increased the average transit time of pellets in the small intestine. The average transit time for the light and heavy pellets being 7 and 25 h respectively. The diameter of pellets is of minor significance. The findings suggest the use of density as a means of modifying the period of absorption of controlled-release pellets.


Dopamine antagonistic effects of a series of analogues of oxiperomide and spiroxatrine measured behaviourally in the rodent. The activity spectra of oxiperomide, spiroxatrine and analogues were determined in two experimental models of abnormal peri-oral movements (induced by intrastriatal dopamine and subcutaneously administered 2-(NN-dipropyl)amino-5,6-dihydroxy-1,2,3,4-tetrahydronaphthalene (NN-diPr-5,6-diOHATN) in the guinea-pig), and in a stereotypy test (induced by subcutaneous apomorphine in the guinea-pig); the ability of the test compounds to induce catalepsy or catatonia in the rat was also determined. The parent compounds oxiperomide and spiroxatrine possessed optimal activity in all tests, although responses to the series of compounds allowed clear differentiation between an ability to antagonize the peri-oral movements (dopamine- or NN-di Pr-5,6-diOHATN induced) and an ability to antagonize apomorphine stereotypy. However, all compounds that antagonized the abnormal peri-oral movements also caused catalepsy/catatonia. The results are considered in terms of the selection of suitable agent(s) for the treatment of peri-oral dyskinesias.


Potentiation of neostigmine and pyridostigmine by 4-aminopyridine in the rat. The interaction between 4-aminopyridine and neostigmine or pyridostigmine was studied in vivo in the rat sciatic nerve-anterior tibialis preparation using the constant infusion of pancuronium technique. The ED50 (dose of drug which produced a 50% antagonism) of neostigmine, pyridostigmine and 4-aminopyridine were 18, 49 and 440 microgram kg(-1) respectively. The addition of 100 microgram kg(-1) of 4-aminopyridine, which produced no antagonism by itself, decreased the neostigmine ED50 to 7.4 microgram kg(-1). The addition of 200 microgram kg(-1) of 4-aminopyridine, which produced a 30% antagonism by itself, decreased the ED50 of pyridostigmine to 11 microgram kg(-1). We conclude that both neostigmine and pyridostigmine interact with 4-aminopyrine synergistically.


The effects of a hydroxyl group on some chemical and biological properties of n-pentyl ammonium salts. The effects of a hydroxyl group on the activity and affinity of compounds related to n-pentyltrimethylammonoum have been studied on the guinea-pig ileum, frog rectus, acetylcholinesterase and on partitioning (Rm) and size (phiov, Vm). The hydroxyl group lowered affinity in all tests, confirming the importance of hydrophobic forces in binding to receptors. Activity on the ileum was lowered appreciably but on the rectus only slightly. The effects on Rm did not indicate any interactions between hydroxyl, onium group and water but the apparent size of the hydroxyl group (deltaphiov) depends on the nature of the onium group.


Autoxidation of polysorbates.

Autoxidation of polysorbates. Aqueous solutions of polysorbate 20 undergo autoxidation on storage, with the peroxide number increasing and subsequently decreasing again, the acidity increasing continuously, the pH and surface tension falling and tending to level off, and the cloud point dropping sharply until turbidity begins at room temperature. The changes are accelerated by light, elevation of temperature, and a copper sulfate catalyst. At the same time, hydrolysis occurs, liberating lauric acid. Analysis of the alterations in these properties leads to the conclusion that hydrolysis has the major influence near room temperature and that oxyethylene undergoes chain shortening at temperatures above 40 degrees. However, evidence of degradation is detectable even in previously unopened commercial samples of polysorbates 20, 40, and 60, warranting attention to the stability of and standards for these surfactants as compared with the solid alkyl ether type of nonionic surfactant.


GI absorption of beta-lactam antibiotics II: deviation from pH--partition hypothesis in penicillin absorption through in situ and in vitro lipoidal barriers. The absorption of propicillin from the rat stomach and small intestine in situ was examined as a function of recirculating solution pH. The in vitro interphase transport from an aqueous buffer of various pH values to the octanol phase was also studied for several penicillins by the use of a two-phase rolling cell. The rate--pH profiles obtained from both in situ and in vitro experiments deviated significantly from the dissociation curves. The degrees of the shifts were approximately 2 pH units for the in situ intestinal absorption of propicillin and in vitro transport of propicillin and cloxacillin, approximately 1.5 pH units for the in vitro transport of penicillin V, and 0.8 pH unit for the in situ stomach absorption of propicillin. These discrepancies from the classical pH--partition hypothesis can be interpreted by the permeation through the lipoidal barrier of the undissociated species of penicillins transported through the aqueous diffusion layer adjacent to the lipoidal surface. All in situ and in vitro experiments tend to support this theory.


Differential pulse polarographic determination of clorazepate monopotassium and dipotassium. Polarographic investigation of clorazepate monopotassium and dipotassium showed two cathodic waves at about -1.28 and -1.66 v. The cathodic wave associated with clorazepate monopotassium or dipotassium at about -1.66 v was a pH-independent, diffusion-controlled wave. This wave was used to develop a specific stability-indicating procedure for clorazepate monopotassium and dipotassium in the presence of their degradation products, namely, nordiazepam, 2-amino-5-chlorobenzophenone, and glycine. The method involves a 10(-2) M LiOH-10(-1) M LiCl extraction of the active ingredient from the formulation, filtration, dilution with the same supporting electrolyte, and then use of the standard addition technique for drug quantitation in capsules. Typical fomulation excipients did not interfere with the analysis. Accuracy and precision of the procedure were 99.55 +/- 0.68%.


Tautomerism of singly protonated chloroquine and quinacrine. Differential absorptiometric pH titrations of the antimalarials chloroquine and quinacrine, employing the respective uncharged species at pH 12.5 in the reference cell, show that protolytic dissociation from the heterocyclic ring nitrogen atoms occurs over pH 6--12. This finding indicates that the singly charged cations of the drugs exist measurably in two tautomeric forms. The tautomeric equilibrium constants were calculated from the titration data. The existence of these tautomeric equilibria may have significance in the pharmacodynamics of chloroquine and quinacrine.


13C-NMR spectra of strychnos alkaloids: brucine and strychnine. The natural abundance 13C-NMR spectra of brucine and strychnine were obtained using the pulse Fourier transform technique. The chemical shifts of various carbon resonances were assigned on the basis of substituent effects on benzene shifts, intesities of signals, multiplicites generated in single-frequency off-resonance-decoupled spectra, and comparisons with the chemical shifts of structurally related compounds.


Loss of hydrophobic amine from solution by adsorption onto container surfaces. The apparent loss of the hydrophobic amine drug alpha-[(dibutylamino)methyl]-6,8-dichloro-2-(3',4'-dichlorophenyl)-4-quinoline-methanol monohydrochloride from solution due to its adsorption onto the surface of its storage container was studied. The drug appeared to be adsorbed only as the free base. Therefore, any perturbations to the solution phase that will help solubilize the drug and thus lower its chemical potential will minimize adsorption. Multilayer drug adsorption to the container surfaces appeared to take place, with some evidence of a highly organized system in the adsorbed phase. Adsorption was minimized when the heterogenous polar functionalities on glass surfaces were covered by a layer of silicone or methacrylate polymer, which yielded less reactive, more hydrophobic surfaces. Loss was also minimized when the environment was kept acidic (pH less than or equal to 4,8), the drug was dissolved in a proton-donating solvent (e.g., chloroform), and an ion-pairing agent (e.g., trichloroacetate) was present to solubilize further the monocationic form of the drug in organic media.


Role of membrane-bound calcium in taste reception of the frog. 1. The frog gustatory responses to various salt stimuli and distilled water were greatly enhanced after the tongue was treated with an alkaline solution containing salts of low concentration. The incubation of the alkali-treated tongue in solution of pH 6.0 containing Ca2+ restored reversibly the behaviour of the gustatory receptor to that before the treatment, while Mg2+ had no ability to do this. 2. The responses to salt stimuli and distilled water were greatly decreased after the tongue was incubated in solutions of pH 5.3 containing Ca2+. 3. On piece of tongue incubated in a solution of pH 5.3 containing 45Ca released a larger amount of 45Ca by the alkali treatment than another piece incubated in pH 7.0. It was concluded that removal of Ca2+ from the gustatory receptor membrane by the alkali treatment led to enhancement of the responses and binding of extra Ca2+ to the membrane by the incubation in acidic CaCl2 solution led to suppression of the responses. We emphasized that a conformational change of the receptor domains plays an important role in the transduction process of the gustatory response.


Evidence for anionic cation transport of lithium, sodium and potassium across the human erythrocyte membrane induced by divalent anions. 1. The passive net transport of Li+ and Na+ across the human red cell membrane was accelerated by the divalent anions carbonate, sulphite, oxalate, phosphite and malonate. Phthalate, maleate, sulphate and succinate were found additionally to stimulate downhill transport of K+. Marked differences in anion efficacy and selectivity were observed. 2. The effects of these 'carbonate type' anions were reversible and fully blocked by SITS, dipyridamole and other inhibitors of anion transfer. 3. Cation transport acceleration induced by the monovalent anions salicylate, benzoate, thiocyanate and 2,4-dinitrophenol were inhibited by dipyridamole, but not affected by SITS. A great number of mono- and polyvalent anions were without detectable influence on Li+ transport. 4. Li+ net uptake induced by oxalate exhibited a pH dependence similar to that reported for halide self exchange. 5. Transport acceleration by carbonate type anions displayed a linear, 1:1 dependence on the concentrations of both the anion and the cation and was symmetric with respect to the two sides of the membrane. 6. It is concluded that the divalent carbonate type anions form singly charged, negative 1:1 ion pairs with the respective alkali metal cations, the ion pairs traversing the red cell membrane via the anion exchange pathway. This concept of anionic formation of some of the ion pairs considered. The relative efficacies and cation selectivities of polyvalent anions can largely be explained on the basis of electrostatic interactions governing ion pair formation. However, the chelating properties, structural flexibility, polarizability of the anions and the accessibility of the ion pairs to the anion exchange pathway need also be considered. 7. An exchange of NaCO-3 ion pairs for internal HCO-3 or Cl- is discussed as a possible mode of cellular pH regulation.


Displacement of activator thresholds in cardiac muscle by protons and calcium ions. 1. The Na current threshold in sheep cardiac Purkinje fibres and in frog atrium is shifted in a positive direction by protons and Ca2+ ions. The titration curves for Purkinje fibres are consistent with a surface potential of -18 mV at pH 7.4 and 1.8 mM-Ca. 2. In Purkinje fibres, the pacemaker K current activation curve, s infinity, is shifted in a positive direction by Ca2+ ions. The results are consistent with a surface potential of -16 mV in normal physiological solutions. 3. The results on s inifinity during pH changes are unexpected. As also shown by Van Bogaert, Vereecke & Carmeliet (1975) the voltage shifts are usually in the opposite direction to that expected from titration of external surface negative charges. 4. Acid solutions reduce the magnitude of iK2 when fully activated. Alkalinity has little effect on iK2. 5. Acidification and alkalinization are both capable of arresting spontaneous activity in Purkinje fibres. The effects of acidity are usually irreversible. The effects of alkalinity are reversible.


The interactions of protons, calcium and potassium ions on cardiac Purkinje fibres. 1. In sheep Purkinje fibres, acidosis shifts the steady-state current-voltage relation in an inward direction over a wide range of negative potentials. The reversal potential for iK2 is shifted in a positive direction. These changes are consistent with K+ accumulation in the extracellular spaces of the Purkinje fibre. 2. These effects of acidosis are completely prevented by increasing [K]0 from the normal range (2.7--5.4 mM) to 10.8 mM. 3. Increasing [Ca]0 from 2 to 6 mM also produces an inward shift in the steady-state current-voltage relation and a positive shift of the iK2 reversal potential. 4. In 10.8 mM-[K]0 these effects are absent even when [Ca]0 is increased to 20 mM. 5. One possible explanation is that K+ ions may protect the Na pump from inhibition by protons and Ca2+ ions. Alternative explanations are also discussed.


Comparison of adenylate cyclase activity and in vitro secretion in the parotid and sublingual glands of the mouse. 1. Adenylate cyclase (EC 4.6.1.1) activity has been determined in the parotid and sublingual glands of the mouse. Optimal activity of the enzyme was obtained at a Mg2+-concentration of 8 mM at pH 8.2, using AMP-PNP as the substrate. 2. Cyclic AMP degradation during the adenylate cyclase assay was relatively high in both the homogenate and the 40,000 g pellet-fraction of the glands. Theophylline was effective in inhibiting this degradation only in the parotid hemogenate, whereas isobutylmethylxanthine inhibited the cyclic AMP degradation in both salivary glands. Using the latter phosphodiesterase inhibitor, we observed a higher adenylate cyclase activity in the sublingual glands than in the parotid glands. 3. Various receptor-selective sympathetic and parasympathetic agonists and antagonists have been tested for their capacity to influence the adenylate cyclase activity and the glycoprotein secretion in the parotid and sublingual glands of the mouse, in vitro. (a) The parotid glycoprotein secretion was increased by beta-adrenergic agonists, which stimulate adenylate cyclase, and by cholinergic muscarinic drugs, which do not activate this enzyme. The adrenergic alpha-agonist phenylephrine appeared to be involved neither in the glycoprotein secretion nor in the direct regulation of the adenylate cyclase activity. (b) The sublingual protein and mucin secretion was increased by cholinergic muscarinic agents. The over-all protein secretion was stimulated also by phenylephrine, but this effect could be blocked by propranolol. The adenylate cyclase activity in membrane preparations was not stimulated by these secretogogues.


Adrenergic and 'non-adrenergic' components in the contractile response of the vas deferens to a single indirect stimulus. 1. The mechanical response of the longitudinal smooth muscle of the rat vas deferens to stimulation of its motor nerves by a single pulse has been examined. The motor nerves were stimulated in vivo via the spinal outflows in the pithed rat or in vitro by field stimulation. 2. The contraction in the whole vas consisted of two components, an initial, rapid, brief contraction reaching a maximum at 300 msec and a second, slower and more prolonged contraction reaching its maximum at 600 msec. When the isolated vas was divided into prostatic and epididymal halves the contribution of these two components varied. The initial rapid component was more prominent in the prostatic half and the slower, second component more prominent in the epididymal half. Lowering the bath temperature caused, in both halves, these two components to merge into a single, slow, prolonged response. Both components were more rapid and briefer than the equivalent response of rat anococcygeus. 3. The second, slow component was abolished by alpha-adrenoceptor antagonist drugs, potentiated and prolonged by drugs which inhibit the neuronal uptake of noradrenaline and absent from tissues taken from rats pre-treated with reserpine, suggesting that the neurotransmitter for this component is noradrenaline. 4. These experiments were extended to the mouse or guinea-pig vas deferens. Both showed the same two component mechanical response as the rat vas and in both the second, slow component was preferentially inhibited by alpha-adrenoceptor antagonists and potentiated by drugs blocking noradrenaline uptake. 5. Drugs known to reduce the response to repetitive nerve stimulation of the vas were examined for their effect on the response to a single stimulus. Lysergic acid diethylamide preferentially inhibited the second, slow phase of contraction whereas apomorphine preferentially inhibited the first rapid phase. Guanethidine inhibited responses but any differential effects could not be analysed due to its stimulant properties. 6. These results show that there are two components even to the response to a single stimulus. The second of these appears to be adrenergic while the transmitter responsible for the first remains to be determined.


Clinical evaluation of oral Candida in cancer chemotherapy patients. No overt clinical oral candidiasis was noted upon either direct oral examination or review of medical records. No significant change in percent of Candida species as compared to other populations is seen in this study. Varying percents of individual species do differ from other reports. Alimentary and digestive tract tumors represented over 56% of the primary malignancies. There was a significant correlation between decrease in salivary pH and increase in percent of Candida species growth. Most significantly, however, the preliminary data in this study might eventually contribute to continuing investigation of Candida species in human cancer patients.


2-Pyrrolidinylideneureas, a new class of central nervous system agents. A series of N-aryl-N'-(1-methyl-2-pyrrolidinylidene)ureas was prepared and screened for pharmacological activity. Congeners possessing either phenyl or phenyl substituted with 4-nitro, 3-bromo, 3-chloro, 3-fluoro, and 3-methyl groups were found to demonstrate anxiolytic activity. 2,6-Disubstitution of the phenyl ring with methyl, chloro, and bromo imparted potent muscle-relaxant properties which appear to be centrally mediated. A significant separation of the anxiolytic and muscle-relaxant properties from other CNS activities, e.g., anticonvulsant, sedative, and hypnotic, was achieved.


2-Methoxyphenylethanolamines, potential beta-adrenergic blocking agents. The effect of the introduction of a 2-methoxy substituent on the beta-adrenergic antagonistic properties of a series of 3- and 4-substituted phenylethanolamines (1) was studied. Both the series of bromo- and methyl-substituted compounds behaved similarly, indicating that electronic forces are not significant in determining beta-adrenergic antagonist activity. When compared with the corresponding phenylethanolamines without a 2-methoxy substitutent, the 2-methoxy-4-substituted derivatives (3a and 3d) had enhanced potency and selectivity but the 2,3- (3b and 3e) and the 2,5-disubstitution patterns (3c and 3f) showed a loss of activity. The inconsistent changes in activity prevented any firm conclusions being made about the effect of the ether oxygen and the beta-adrenoceptor antagonistic activity of phenoxypropanolamines.


Derivatives of 1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4]benzodiazepine-5,11(10H)-dione as anxiolytic agents. A study of the pharmacological properties of pyrrolo[2,1-c][1,4]benzodiazepine derivatives led to the choice of (+)-1,2,3,11a-tetrahydro-10-methyl-5H-pyrrolol[2,1-c][1,4]benzodiazepine-5,11)10H)-dione as a candidate for anxiolytic evaluation in a limited clinical trial in man. Metabolism studies in laboratory animals have pointed to rapid hydroxylation, possibly in the 3 and 11a positions. A series of compouds containing methyl groups in one or more of these positions has been prepared in an effort to block metabolism and thereby obtain more active or longer acting compounds. All of these derivatives were less active than the parent compound.


Psychotropic agents. 3. 4-(4-Substituted piperidinyl)-1-(4-fluorophenyl)-1-butanones with potent neuroleptic activity. A series of 1-(4-fluorophenyl)-4-(1-piperidinyl)-1-butanones substituted with benzimidazole, benzotriazole, or quinoxaline at the 4 position of the piperidine ring was synthesized and subjected to neuroleptic tests. Neuroleptic activities of several compounds were comparable to those of haloperidol. In particular, 4-[4-(2,3-dihydro-2-thioxo-1H-benzimidazol-1-yl)-1-piperidinyl]-1-(4-fluorophenyl)-1-butanone (10) was characterized by having a potent neuroleptic activity with less liability to the extrapyramidal side effect.


Synthesis of spiro[isobenzofuran-1(3H),4'-piperidines] as potential central nervous system agents. 4. Central nervous system depressants. The synthesis of 1'-[3-(4-fluorobenzyoyl)propyl]-3-phenylspiro[isobenzofuran-1(3H),4'-piperidine] (2a) and eight halo and methoxy analogues is described. The compounds were generally more potent per os than chlorpromazine in the Sidman avoidance paradigm in rats and less potent than haloperido. 1'-[3-(4-Fluorobenzoyl)propyl]-3-(4-fluorophenyl)spiro[isobenzofuran-1(3H),4'-piperidine] (2e) approached the per os potency of haloperidol in this test and was shown to be active in inhibiting monkey avoidance also. Compound 2e was much less active than haloperidol in antagonizing apomorphine-induced emesis in dogs, apomorphine-induced stereotypy in rats, and amphetamine-induced circling in lesioned rats. This lack of nonselective, dopamine-receptor blocking effects makes 2e attrative as a potential neuroleptic.


Synthesis and beta-adrenoceptor activity of the erythro and threo isomers of substituted alpha-hydroxytrimetoquinol. The synthesis and pharmacological activity of erythro and threo isomers of 1-(3',4',5'-trimethoxy-alpha-hydroxy-benzyl)-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline, 2 and 3, are reported. The structural assignments of 2 and 3 are based upon NMR spectra of the 6,7-dibenzyl precursors, 6 and 10, and of the synthetic derivatives of 13alpha- and 13beta-hydroxy-2,3-(dibenzyloxy)-9,10,11-trimethoxytetrahydroprotoberberine, 8 and 12, respectively. The erythro isomer 2 was a more potent beta-adrenoceptor stimulant than the threo isomer 3 in guinea pig atrial, guinea pig tracheal, and rat adipocyte preparations. The differential activity of these compounds on lipolysis was favorably correlated to changes in the stimulation of adenylate cyclase activity and cAMP accumulation in rat adipocytes.


Neuroleptics related to butaclamol. An investigation of the effects of chlorine substituents on the aromatic rings. The synthesis of analogues of the antipsychotic drug butaclamol bearing chloro substituents on the benzene rings is described. On the basis of a perceived topographical similarity of a putative chlorophenylethylamine pharmacophore present in these analogues and in VUFB-10032 and doclothepin, agents related to octoclothepin which do not induce catalepsy, they have been tested for "noncataleptic" neuroleptic activity. None of the butaclamol analogues exhibit this type of activity. Depending on the position of the chlorine, the analogues either retained butaclamol-like activity or were inactive.


(2-exo-3-endo)-2-Aryltropane-3-carboxylic esters, a new class of narcotic antagonists. Tropanes 4 bearing an exo aromatic group on carbon-2, an endo-carbomethoxy group on carbon-3, and either a methyl group or a hydrogen on the nitrogen were found to be narcotic antagonists which were devoid of demonstrable analgesic activity. The activity resided in the 1S enantiomer. Compound 4 (R = m-hydroxyphenyl) showed an AD50 of 0.37 mg/kg sc and 1.8 mg/kg po (rats) as an antagonist in the Harris-Pierson modification of the D'Amour-Smith test. The tropane esters for this study were prepared by a Grignard reaction which gave essentially complete 1,4-addition in the absence of copper salts. Nearly equal quantities of esters epimeric at carbon-3 were formed.


(exo, exo)-2-Aryltropane-3-carboxylic esters, hypoglycemic agents with accompanying analgesic activity. (exo, exo)-2-Aryltropane-3-carboxylic esters of types 6, 7, and 10 lower circulating blood glucose levels by 60--80%. This activity is accompanied by an analgesic activity roughly equal to that of codeine. Both of these activities reside in the 1R enantiomer and extensive structure-activity studies failed to separate them. The specific opioid antagonist nalorphine blocks the analgesic activity but does not diminish the hypoglycemic action. Conformational integrity afforded by the ethylene bridge is neccessary for the observed activities.


Pharmacology of some metabolites of triazolam, alprazolam, and diazepam prepared by a simple, one-step oxidation of benzodiazepines. A simple, one-step chemical oxidation of triazolam (7) to its 4-hydroxy analogue, 7a, has been developed and applied to other triazolo- and imidazobenzodiazepines. The reaction may be used to convert diazepam to temazepam. 4-Hydroxytriazolo[4,3-a][1,4]benzodiazepines have low central nervous system sedative and anticonvulsant activity in sharp contrast to metabolites of diazepam which remain active. While 10, an alpha-hydroxy metabolite of triazolam, retains much of the activity of 7, 10a, and alpha,4-dihydroxy metabolite of triazolam, is 250 times less potent than 7 on the nicotine-antagonism assay and over 300 times less potent on the traction assay.


Stereoelectronic factors in the binding of substrate analogues and inhibitors to purine nucleoside phosphorylase isolated from human erythrocytes. Several aspects of the stereoelectronic requirements of substrates of human erythrocytic purine nucleoside phosphorylase (E.C. 2.4.2.1) were elucidated providing the following information: (a) the N1 position cannot have a nonhydrogen substituent; (b) the 5'-OH group must be present for catalytic activity to be exhibited but is not an essential functional group for inhibitory action to be observed; (c) on the C8 position groups larger than -NH2 or -Br cannot be accommodated; (d) the syn-glycosyl conformation (i.e., 8-bromoguanosine) is acceptable but may not be an absolute requirement for phosphorolysis; (e) among nucleic base inhibitors methylation at N3, N7, or N9 vastly decreases the inhibitory properties as does a nitrogen in lieu of C-H in the 8 position. The results clearly indicate that this enzyme differs in its stereoelectronic requirements from the Escherichia coli enzyme.


Nondepressant beta-adrenergic blocking agents. 1. Substituted 3-amino-1-(5,6,7,8-tetrahydro-1-naphthoxy)-2-propanols. A series of 3-amino-1-(5,6,7,8-tetrahydronaphthoxy)-2-propanols was synthesized and investigated for beta-adrenergic blocking activity and direct myocardial depressant action. The cis- and trans-diols 12--15 were found to retain the beta-blocking potency of propranolol but to lack its myocardial depressant action. Compound 15 (nadolol) is currently undergoing extensive clinical evaluation as a potential antianginal, antiarrhythmic, and antihypertensive agent.


Structure-activity study of beta-adrenergic agents using the SIMCA method of pattern recognition. The SIMCA method of pattern recognition (PaRC) was used to analyze structure-activity data for a series of phenethylamine agonists and antagonists of the beta-adrenergic receptor. On the basis of physicochemical substituent parameters the SIMCA method classified correctly 100% of the agonists and 88% of the antagonists. In addition, parameters derived from the class models were correlated with the biological activities of the agonists and antagonists, respectively. Test compounds not included in the initial data analysis were classified and their activities estimated. The applicability of pattern recognition in structure-activity studies in general is discussed.


Synthesis and biological evaluation of some 2-amino-4-aryl-3H-1,5-benzodiazepine analogues of clozapine. 2-Amino-4-aryl-3H-1,5-benzodiazepines were prepared and evaluated for potential neuroleptic activity. Compound 60 showed some activity in the four assays; however, the activity was not consistently observed among other members of the series. The data reflected that the structural modifications led to a decrease in activity relative to clozapine. It was apparent that the 1,5-benzodiazepine portion of clozapine is not responsible for its antipsychotic activity.


Cardioselectivity as a function of molecular structure in beta-adrenoceptor blocking agents of the 1-(para-substituted aryloxy)-3-(isopropylamino)propan-2-ol type. The relationship between molecular structure and cardioselectivity is described in the 1-(para-substituted aryl-oxy)-3-(isoprophylamino)propan-2-ol type of beta-adrenoceptor blocking agents. Cardioselectivity in the aforementioned series requires that the aromatic substitution in the position para to the amino alcohol side chain will have a minimal linear length of 5.0 A. Highest cardioselectivity is obtained when this para substituent is a rigid group coplanar with the aromatic ring. This may result from steric hindrance for binding at the beta2-adrenoceptor subtype which does not occur in the beta1 subtype. Evidence in favor of this suggestion was obtained by the finding that the trans isomer of 1-[4-(1-propenyl)-2-methoxyphenoxy]-3-(isopropylamino)propan-2-ol is cardioselective (beta1/beta2 = 25), whereas the cis isomer is beta2 selective (beta1/beta2 = 0.1).


HCN: a plausible source of purines, pyrimidines and amino acids on the primitive earth. Dilute (0.1 M) solutions of HCN condense to oligomers at pH 9.2. Hydrolysis of these oligomers yields 4,5-dihydroxypyrimidine, orotic acid, 5-hydroxyuracil, adenine, 4-aminoimidazole-5-carboxamide and amino acids. These results, together with the earlier data, demonstrate that the three main classes of nitrogen-containing biomolecules, purines, pyrimidines and amino acids may have originated from HCN on the primitive earth. The observation of orotic acid and 4-aminoimidazole-5-carboxyamide suggests that the contemporary biosynthetic pathways for nucleotides may have evolved from the compounds released on hydrolysis of HCN oligomers.


Renin profiling for diagnosis and treatment of hypertension. Recent research shows that the renin-angiotensin-aldosterone axis either maintains or causes some or all of the high blood pressure of most patients and demonstrates anew that renin-sodium profiling defines this involvement. Performed with a serum potassium measurement, this now reliable test is useful for primary screening and then, in conjunction with renal vein renin studies or an aldosterone profile, for diagnosis or exclusion of surgically curable renovascular or adrenocortical hypertensions. For the remaining majority with essential hypertension, renin profiling exposes the relative participation of either vasoconstriction or volume factors, thereby guiding simpler, more specific, and predictably effective antirenin or antivolume treatments. Renin profiling identifies those in whom treatment should begin with a beta-blocker as opposed to a diuretic while not infrequently also providing baseline information about severity and prognosis in individual patients.


Enzymatic decarboxylation of L-threo-3,4-dihydroxyphenylserine in rat heart. Decarboxylation of L-threo-3,4-dihydroxyphenylserine (L-threo-DOPS) by the higher speed supernatant of the rat heart homogenate and the regional distribution of L-threo-DOPS decarboxylase activity were examined. Decarboxylation was demonstrated to occur specifically with L-isomer but not with D-isomer. Addition of pyrogallol was necessary for maximal recovery of norepinephrine. The optimal condition for decarboxylation of L-threo-DOPS by the rat heart enzyme was similar to conditions required with the enzymes from brain and kidney. Under the optimal conditions, Km and Vmax for L-threo-DOPS were 2.1 mM and 6.4 nmoles/mg protein/15 min, respectively. Decarboxylation of L-threo-DOPS  was markedly inhibited by D-threo-DOPS and D-DOPA. The L-aromatic amino acid decarboxylase activity was highest in the right auricle followed by the atrial body, the left auricle, the right ventricle and the left ventricle.


Gastric cancer in Colombia. IV. Nitrite and other ions in gastric contents of residents from a high-risk region. Samples of gastric contents from 2 groups of patients from a region of high risk for gastric cancer were analyzed for pH, nitrite, nitrate, thiocyanate, and chloride. In each group, the patients could be divided into 2 subgroups: those with a gastric pH of less than 5 and those with a gastric pH of greater than 5. Above pH 5, nitrite was correlated with nitrate. The pH greater than 5 subgroups had significantly higher (P less than 0.01) nitrite content (20- to 100-fold). Some high- and low-nitrite samples were also analyzed for macro and trace metal ions, but differences were not significant. This is the first report in which patients with diagnosed gastric pathology related to a precancerous state were shown to have high levels of a putative carcinogen precursor. The results were compatible with our original hypothesis of intragastric nitrite formation by bacterial reduction of nitrate and concomitant synthesis of carcinogenic N-nitroso compounds.


Fulminant pneumococcal bacteremia in an asplenic chronic hemodialysis patient. A chronic hemodialysis patient, previously splenectomized because of trauma in conjunction with gastrectomy, developed bacteremia with type 18 Streptococcus pneumoniae and died within 13 hours of onset of symptoms. Characteristics of this illness were severe hypoglycemia, pneumococci visible on peripheral blood smear, disseminated intravascular coagulation, neutropenia, and in vitro hemolysis. Splenectomy should be considered with caution in uremic patients and in renal transplant recipients because of the increased risk of fulminant bacteremia. Polyvalent pneumococcal vaccine may be helpful in preventing this syndrome in such asplenic patients.


[Use of functional loads in assessing the state of the humoral systems in arterial hypertension]. Some methodical approaches in examination of patients for the "hormonal profile" in 50 healthy individuals and in 100 patients with hypertensive disease an attempt is made to standardize the conditions for conducting the tests and the appraisal of their results. On the grounds of the study of the prostaglandin content, activity of blood plasma renin, and excretion of kallikrein in the urine in different types of loads it is concluded that it is necessary to choose an adequate load for each humoral system; when analysing the results of the load tests one must take into account the initial condition of the humoral system under study since the reaction of the system to the load depends on it.


[State of the sympathetic-adrenal system and renin activity in the blood in children with primary arterial hypertension in the process of beta-blockader treatment]. The condition of the sympatheticoadrenal system and blood renin activity were studied in healthy children and in children suffering from primary arterial hypertension and treated with beta-blocking agents (Obzidan, Visken). A hypotensive effect was noted in the group of patients suffering from primary arterial hypertension with a high blood renin level and increased excretion of catecholamines in the urine.


[General hemodynamics and kidney function in hypertension in pregnant women]. The indices of general hemodynamics and renal function were studied in 362 pregnant women (healthy and those suffering from hypertensive disease, glomerulonephritis or nephropathy) and in 20 healthy but not pregnant women. It was found that the hypokinetic type of circulation developed in glomerulonephritis and nephropathy in pregnant women and that the hyperkinetic type changes to eu- and hypokinetic types in pregnant women suffering from hypertensive disease. In all variants of hypertension in pregnancy, the renal blood flow and glomerular filtration reduce, while the resistance to the blood flow in the vessels of the kidneys increases. The role of the sympathetico-adrenal and the renin-aldosterone systems in the changes in hemodynamics and renal function in healthy and sick pregnant women was studied. An opinion is voiced that increased sensitivity of the vascular receptors to vasoactive substances is very important.


Semen preservation in Macaca fascicularis. Semen was collected from adult male Macaca fascicularis using a rectal probe for electro-ejaculation. The effect on sperm motility of varying semen extender egg yolk concentration, pH, glycerol concentration, and equilibration times of sperm with glycerol was examined. No significant difference was observed between motilities at extender egg yolk concentrations of 10% to 40%. Progressive motility was significantly greater at pH 7.2 and 8.0 than at 5.8, 6.5, and 8.7 (p less than 0.05). Glycerol concentrations of 7% and 10% yielded optimum progressive motility after freezing. A 1-minute equilibration of semen in extender containing glycerol resulted in greater sperm motility after freezing than did equilibration for 25 or 45 minutes.


Beta-adrenergic receptors: regulatory role of agonists. Direct radioligand binding studies have been used to probe the molecular mechanisms whereby agonist catecholamines regulate the function of beta-adrenergic receptors in a model system, the frog erythrocyte. The unique characteristics of agonist as opposed to antagonist action are first, the ability to stimulate the adenylate cyclase through the receptor and second, the ability to desensitize the system by alterations induced in beta-adrenergic receptors. These properties of agonist are not shared by antagonist despite the high affinity and specificity of antagonist binding to the beta-adrenergic receptors. Agonist and antagonist receptor complexes may be distinguished in a variety of ways including differences in their sensitivity to regulatory guanine nucleotides and also by gel chromatography on AcA 34 Ultragel. The agonist receptor complex appears to elute from the columns with an apparently increased size. A "dynamic receptor affinity model" of beta-adrenergic receptor action is proposed which features several distinct conformational states of the receptor. Agonists have much higher affinity for the physiologically active or coupled state of the receptor, whereas antagonists have equal affinity for both. In addition, a third "desensitized" state of the receptor is also postulated to exist.


Indomethacin stimulation of lipid peroxidation and chemiluminescense in rat liver microsomes. Peroxidation of endogenous lipid by liver microsomes, coupled with oxidation of reduced nicotinamide adenine dinucleotide phosphate (NADPH) and measured as thiobarbituric acid reactive materials, is markedly stimulated in the presence of indomethacin [1-(p-chlorobenzyl)-5-methoxy-2-methyl-3-indole acetic acid] (0.1--1.0 mM). Concurrently, indomethacin enhances the lipolysis of membrane phospholipid containing arachidonic acid but has no effect on the rate of O2 uptake in these samples. The system generates a rapidly developed chemiliminescense (CL), the intensity and rate of development of which are related to indomethacin concentration. The microsomal CL generated in the presence of indomethacin is distinct from the previously reported CL in that the time required for maximum intensity development is a matter of seconds (20--180) rather than hours. The enhanced CL is believed to be due to an energy transfer reaction whereby a high energy species transfers energy to the indomethacin molecule, which, in turn, decays via chemiluminescense. An enhanced chemiluminescense was also observed when indomethacin was added to a lipoxidase system and superoxide generating system (axanthine oxidase). Based on inhibitor studies, the rapidly developed chemiluminescense of the microsomal system requires cytochrome P-450 in addition to NADPH and coordinated iron ions. The results indicate that the CL is related to neither hydroxyl free radical nor superoxide anion formation.


Effect of catecholamines and beta-blockers on linoleic acid desaturation activity. The effect of catecholamines and adrenergic blocking agents on the oxidative desaturation of linoleic acid in rat liver microsomes was studied. Epinephrine (1 mg/kg/body weight) produced a significant decrease on the conversion of [1-14C]linoleic acid to gamma-linolenic acid. The effect of epinephrine was blocked by single injections of the beta blockers propranolol (10 mg/kg body weight) or dichloroisoproterenol 30 min before the hormone treatment. Isoproterenol (100 microgram/kg body weight) produced a significant decrease on the activity of the linoleyl-CoA desaturase. The effect of the catecholamines was postulated to be mediated through beta receptors by an enhancement of the intracellular levels of cyclic AMP.


[The efficacy of a maintenance dose of 100 mg doxycycline (Vibramycin) daily in bacterial infections of chronic bronchitis (author's transl)]. The efficacy and tolerance of a daily maintenance dose of 100 mg doxycycline (Vibramycin) was investigated in ambulant patients with chronic obstructive pulmonary disease (COPD) and bacterial infection by an open study with clinical, bacteriological and cytological control. The dosage was well tolerated and effective in the elimination of H. influenzae and pneumococci. A higher maintenance dose seems not to be necessary for this type of infections.


[Treatment of chronic bronchitis with Supristol and a cotrimoxazole preparation--comparative bacteriological and clinical findings, results of a double-blind study (author's transl)]. In a randomized double-blind study on the treatment of chronic bronchitis, Supristol (co-trifamole) was compared with co-trimoxazole. 163 patients participated in the clinical trial that was performed at three different hospitals in accordance with a uniform trial plan. The microbiological investigations were done centrally at a hygiene institute for all three hospitals. The bacteriological results from the 61 patients whose antibiogram revealed sensitive pathogens before starting the therapy demonstrate the equal effectivenesss of the two preparations. With Supristol there was an elimination rate of 95.3%, and with co-trimoxazole of 93.2%. The clinical parameters do not show any difference in effectiveness either.


Fatty acid biosynthesis in yeast.

Fatty acid biosynthesis in yeast. Fatty acid synthetase and acetyl CoA carboxylase mutants have been used to study several aspects of fatty acid biosynthesis in yeast: the contribution of the various enzymes of fatty acid biosynthesis and modification to the overall cellular fatty acid composition, the mechanism of fatty acyl chain elongation in yeast, the molecular structure and the reaction mechanism of the fatty acid synthetase complex and the genetic control of the biosynthesis of this multi-enzyme system. Genetic and biochemical evidence suggest an alpha6beta6 molecular structure of this complex, where alpha and beta are multifunctional proteins comprising, respectively, 3 and 5 of the various fatty acid synthetase component functions. The two subunits alpha and beta are synthesized on two different, unliked genes, fas 2 and fas 1. The biosynthesis of both is coordinated. The various component enzyme activities reside in distinct domains on the multifunctional chains. While most domains appear to be functionally independent, the three acyl transferases exhibit extensive mutual interactions. It is suggested that the biosynthesis of a multifunctional protein is favoured on the grounds of kinetics and regulation as compared with the formation of a complex of the corresponding individual enzymes.


[A peculiar type of chronic persisting hepatitis (author's transl)]. Six cases of persisting hepatitis with a duration of 3 to 9 years are reported. During this time 2--3 acute inflammatory episodes occurred with transaminase levels raised to more than 500 U/l. Icterus did not occur during these attacks. The remaining laboratory parameters which are usually normal in persisting hepatitis (gamma-GT, bromsulfalein) were pathological during the acute attack and later became normal. As the disease progressed after overcoming the attack, the transaminases were only slightly raised in all cases. Laparoscopy and histological examinations of between three and nine punctates gave an unchanged picture of chronic persisting hepatitis. During the acute attack the inflammatory activities in the liver punctates increased, there were also increased single cell necroses. It is suggested that this peculiar type of chronic persisting hepatitis shall be called "acute recurrent persistent hepatitis.".


[Separation of potassium and calcium channels in the nerve cell soma membrane]. Calcium inward and potassium outward currents were studied on internally dialysed isolated neurons of the snail Helix pomatia. Different sensitivity of the corresponding channels to changes in external pH was found. This difference was used for separation of their activation regions on the potential axis so that the characteristics of the inward and outward currents could be studied with minimal overlap. It is shown that the outward current channels possess a definite permeability to Tris ions (PTris :PK=0.05). This explains the impossibility to switch off this current by substituting Tris for internal potassium. The channels for the inward calcium current inactivate slowly with a first order kinetic; their instantaneous current-voltage characteristic reveals considerable Goldman-type rectification. The selectivity of the calcium channels to other bivallent cations is Ba:Sr:Ca:Mg=2.8:2.6:1.0:0.2.


[A case of Churg-Strauss granulomatous angiitis: an autonomous clinical entity or a variant of panarteritis nodosa?]. A fatal case of Churg and Strauss granulomatous angiitis in a 21-yr-old woman is described. Its unusual features included marked personal and familial dysreaction, sudden onset, bronchopulmonitis with distinct eosinophilia and antibiotic- and cortisone-resistant pericarditis, and a terminal picture of generalised septico-pyaemia. The histological findings included necrotising systemic vasculitis with granulomas, some of them extravascular, in various organs. Differential diagnosis and the relation between this form and panarteritis nodosa are discussed.


Self-poisoning in Auckland reconsidered. Data is presented describing 345 self-poisoners presenting at the accident and emergency department and 211 referred to the psychiatric liaison service at Auckland Hospital during the 12 month period between August 1976 and July 1977. Both groups show the age and sex distribution commonly seen in self-poisoning, with a preponderance of young females. Referral rates increased with age and with the implication of anti-depressants, neuroleptics and anticonvulsants. Only 20 percent of the referred patients had a serious psychiatric disorder. Personality disorder, depressive neurosis and transient situational disturbance accounted for 80 percent. The benzodiazepine group of anxiolytics was implicated in 40 percent of incidents and the use of this class of medication is discussed in relation to the problem of self-poisoning. It is suggested that if all Auckland doctors made a decision not to prescribe oral benzodiazepines, the incidence of self-poisoning in the city could be reduced to a half to two thirds of its present proportions.


Continuous fetal tissue pH measurement in labor. Fifty-one women in labor had continuous monitoring of fetal scalp tissue pH, fetal heart rate by ECG, and uterine contractions. A miniature pH electrode secured by a double spiral fetal ECG electrode was used for measurement of fetal pH every 15 seconds. The results were correlated with fetal scalp blood pH values obtained simultaneously. Fetal scalp sampling is intermittent, requires repeated scalp incisions, is subject to errors due to air mixing and coagulation of the blood sample, and is uncomfortable for the parturient. Placement of the tissue pH electrode allows continuous data recording with the minimum discomfort to the patient and the least number of fetal scalp incisions. Clinical use of the tissue pH electrode might be a practical alternative to fetal scalp samples, if the data obtained accurately reflect fetal status.


Bupivacaine paracervical block. Effects on the fetus and neonate. To evaluate effects on the fetus and neonate, 53 paracervical blocks were administered to 38 low-risk parturients using a controlled superficial injection of 0.25% bupivacaine. Continuous fetal heart rate monitoring revealed no instance of bradycardia or late deceleration pattern. Apgar scores, cord arterial and venous pH values, and neurobehavioral evaluations of the neonates were similar to those observed in other low-risk patients. The effectiveness of pain relief of the blocks was less than expected or reported by other authors. Although we observed no harmful effects on the fetus or neonate from the superficial paracervical injection of bupivacaine, we did not find this combination of drug and technique to give dependable, effective pain relief in labor.


Neonatal acid-base balance in spontaneous and instrumental vaginal deliveries. The acid-base balances of 63 neonates delivered either spontaneously or by the vacuum extractor or low forceps were compared. The outcome was similar in neonates delivered by vacuum extractor or forceps. However, significant differences were noted in the pH and base deficit of infants born by instrumental versus spontaneous delivery. These differences were no longer present when groups with similar duration in the second stage of labor were compared. We conclude that the use of instruments for outlet vaginal delivery carries no additional risk for the fetus.


The effect of nicotine on fetal breathing movements in conscious pregnant ewes. Nicotine (0.14--0.25 mg/kg), injected intravenously or intraarterially into conscious pregnant ewes, caused a decrease in fetal PaO2 within 5 minutes, persisting for up to 30 minutes. There was a significant fall in the incidence of fetal breathing movements. These changes did not occur if the ewe was treated with an alpha-blocking agent (phentolamine) or if the nicotine was infused for 30 minutes at 0.27 to 0.85 mg/minute. Nicotine crossed the placenta; fetal concentrations equaled those in the ewe 5 minutes after the injection and remained at or above maternal levels for 1 hour. Nicotine given directly to the fetus (0.005--0.03 mg/kg estimated fetal weight) stimulated fetal breathing movements in a dose-related manner. We suggest that the maternal injection of nicotine results in a fall of uterine blood flow by a sympathomimetic action, leading to transient fetal hypoxemia and a reduction of fetal breathing movements and that a similar phenomenon may occur when a pregnant woman smokes cigarettes.


Inhibition of gluconeogenesis in isolated rat kidney tubules by branched chain alpha-ketoacids. Isolated rat kidney tubules served as a model to investigate the direct effects of branched chain aminoacids, their alpha-ketoderivatives, and of the homolog straight chain aliphatic alpha-ketoacids on renal gluconeogenesis. It is demonstrated that the alpha-ketoderivatives, rather than the branched chain aminoacids themselves, are potent inhibitors of renal gluconeogenesis from precursors, entering the glucogenic pathway on all levels below and above triose phosphate. This inhibitory action is not specific for the branched chain alpha-ketoacids, since it is also observed in the presence of the homolog straight chain aliphatic alpha-ketoacids. The suppression of renal gluconeogenesis by alpha-ketoacids can not be explained by a direct inhibition of gluconeogenic reactions, by inhibition of cellular respiration, or by interference with the stimulatory action of Ca++, cAMP, and L-lysine on renal gluconeogenesis. Although the point of inhibitory attack of alpha-ketoacids in renal gluconeogenesis could not be localized, an impairment of the kidney to respond to metabolic acidosis with an increase of gluconeogenesis was observed, since the pH optimum of renal gluconeogenesis was shifted from pH 6.8 to pH 7.7 in the presence of alpha-ketoisovaleric acid.


Pneumococcal polysaccharide immunization in infants and children. By using indirect hemagglutination, the antibody responses of normal infants and children to an octavalent pneumococcal vaccine that contained pneumococcal polysaccharide types 1, 3, 6, 7, 14, 18, 19, and 23 were evaluated. By 2 years of age, there was a significant rise in hemagglutination titers to all the polysaccharide types, except type 19. By 6 to 8 months of age, five of the eight types of pneumococcal polysaccharides tested resulted in up to 60% responders and, by 2 years, a significant number responded to all pneumococcal polysaccharide types in the vaccine. Pneumococcal polysaccharide type 3 resulted in a significant antibody response as early as 3 months of age, whereas type 19 never resulted in a significant antibody response. Except for type 3, it seemed that when the other pneumococcal polysaccharides tested produced an antibody response, the degree of resonse did not subsequently change significantly with increasing age. The relationship of antibody response to age for pneumococcal polysaccharides is similar to that found for other polysaccharide vaccines. Based on the results of our study, we would recommend immunization with pneumococcal vaccine at 6 months of age with repeat immunization at 2 years of age, especially in high-risk children.


Salicylate and gastric mucosal damage.

Salicylate and gastric mucosal damage. The mechanism(s) of salicylate-induced damage of the gastric mucosa is complex. The presence of acid in the lumen is essential for the occurrence of such injury. Although the absorption of salicylate is greater in the presence of acid, salicylate can selectively increase cation permeability both in the absence and presence of acid. Recent studies suggest that this permits increased diffusion of luminal acid into the tissue, which leads to major permeability changes of the mucosa. Salicylate also affects metabolic processes of the gastric mucosa, which appear to be independent of the increased diffusion of acid into the tissue also caused by salicylate. The release of histamine, which has been shown to occur in vivo as a result of mucosal damage, does not appear to intensify existing mucosal injury in an isolated system.


Phosphate transport in the proximal convolution of the rat kidney. III. Effect of extracellular and intracellular pH. Inorganic phosphate (Pi) transport was evaluated using the standing droplet method with simultaneous microperfusion of the peritubular capillaries. To evaluate rather small differences in Pi transport and to eliminate the influence of tubular heterogeneity, the technique of crossed paired samples was applied. 1. In chronic PTX rats changing the luminal or both luminal and peritubular pH by varying the HCO-3-concentration between 4 and 50 mmol/l at constant 5% CO2 had no influence on Pi transport. 2. If, however, bicarbonate was omitted from the perfusate and 2 mmol/l phosphate (pH 7.4) was the only buffer, Pi transport was decreased from the control. It was, however, further reduced when the perfusates were gased with 5% CO2 i.e. the starting pH was 5.6. 3. When the solutions contained HEPES buffer (25 mmol/l), Pi transport at pH 8 was much larger than at pH 6.0. 4. Raising the CO2 pressure from 35 to 70 mm Hg did not change the Pi transport when both perfusates had a HCO-3-concentration of 25 mmol/l. It reduced, however, the Pi transport, when the luminal perfusate had only 4 mmol/l bicarbonate. 5. Lowering the CO2 pressure from 38 to 7.6 mm Hg did hardly change the Pi transport when the luminal perfusate contained 4 mmol/l bicarbonate. It lowered, however, the Pi transport significantly when the luminal perfusate had 2k mmol/l bicarbonate. 6. Acetazolamide, 10-4M, lowered the Pi transport when the luminal perfusate contained 4 or 25 mmol/l bicarbonate. At 4 mmol/l luminal HCO-3, raising the pCO2 to 228 mmol/l depressed Pi transport even more. At 25 mmol/l luminal bicarbonate, raising the pCO2 from 38 to 114 mm Hg reversed the acetazolamide inhibition of the Pi transport almost completely. The data indicate that luminal acidosis and intracellular alkalosis inhibits the transtubular Pi transport. A shift of the intracellular pH to a more alkaline value seems to be responsible for the inhibition of Pi transport by acetazolamide, while omission of buffer from the perfusate inhibits Pi transport by effecting an acidic luminal pH.


[Interaction of labetalol with alpha- and beta-adrenergic receptors : biochemical study (author's transl)]. The interaction of a new antihypertensive agent, labetalol was demonstrated in subcellular preparations enriched in alpha- and beta-adrenergic receptors. This result give direct confirmation of pharmacological data observed in isolated organs and in vivo. It illustrates the value of biochemical studies of receptors in order to define the mechanism of action of drugs.


A comparison of the circular dichroism spectra of synthetic DNA sequences of the homopurine . homopyrimidine and mixed purine- pyrimidine types. We have obtained the ultraviolet circular dichroism spectra of two repeating trinucleotide DNAs, poly [d(A-G-G).d(C-C-T)] and poly[d(A-A-G).d(C-T-T)], that have all purines on one strand and all pyrimidines on the other. These spectra, together with spectra of other synthetic polymers, can be combined to give 3 first-neighbor calculations of the spectrum of poly[d(A).d(T)] and 2 first-neighbor calculations of the spectrum of poly [d(G).d(C)]. The results show (1) that first-neighbor calculations utilizing only spectra of homopurine.homopyrimidine DNA sequences are no more accurate than are similar calculations that involve spectra of mixed purine-pyrimidine sequences, demonstrating that double-stranded homopurine.homopyrimidine sequences do not obviously belong to a special class of secondary conformations, and (2) that the wavelength region above 250 nm in the CD spectra of synthetic DNAs is least predictable from first-neighbor equations, probably because this region is especially sensitive to sequence-dependent conformational differences.


Fluorescent complexes of DNA with DAPI 4',6-diamidine-2-phenyl indole.2HCl or DCI 4',6-dicarboxyamide-2-phenyl indole. 4',6-Dioarboxyamide-2-phenyl indole (DCI), a non-ionic structural analogue of 4',6-diamidine-2-phenyl indole.2HCl (DAPI), was synthesized in order to verify the hypothesis of intercalation of both dyes into the DNA double helix. The influence of pH, viscosity, and different concentrations of SDS (sodium dodecylsulphate) or NaCl on the optical and fluorescent properties and the changes in thermal transition of both dye complexes with DNA confirm the affinity of the dyes to the double helix as well as their stabilizing influence on the secondary DNA structure. The results of binding studies, carried out by fluorescent methods have shown that the dyes are strongly bound to DNA, though the number of binding sites is small. According to the experimental data, the fluorescent properties of DAPI and DCI complexes with DNA are connected with the intercalating binding mechanism of these dyes. On the other hand, the eventual ionic or hydrogen bonds of dyes outside the DNA helix do not change noticeably their fluorescent properties.


Phosphorus-31 NMR studies of E. coli ribosomes. Phosphorus-31 nuclear magnetic resonance spectra, relaxation times and nuclear Overhauser (NOE) enhancement have been measured for E. coli ribosomes, subunits and rRNA. NOE and T1 experiments reveal that the phosphorus relaxation in this organelle is largely dipolar in origin. Moreover these results imply the presence of internal motion within the RNA chain with a correlation time of about 3-5 x 10(-9) sec. In all cases the predominant resonance is centered at about -1.5 ppm (relative to 85% H3PO4) as expected for a phosphodiester linkage where there is a large degree of double helix. The linewidth narrows by about a factor of four when the ribosomal proteins are removed indicating a substantial immobilization of the RNA when it is assembled into the ribosome. In addition to the phosphodiester resonance, ribosomes also reveal one or two narrower resonances shifted to low field by 1-4 ppm. Based on the observation that these resonances show a pH dependent chemical shift, we assign them to phosphate monoesters i.e. terminal 3' or 5' phosphate groups. These terminal phosphates are due to short oligomers of RNA derived from the terminus of the chain.


Nucleosome conformation: pH and organic solvent effects. Monomer nucleosomes (nu 1) from chicken erythrocyte nuclei were examined in aqueous buffers (8 greater than pH greater than 3) and in solvent mixtures (i.e., water and ethanol, ethylene glycol, dioxane, dimethyl sulfoxide, 2-methyl-2,4-pentanediol, polyethylene glycol, sucrose, or urea). Circular dichroism, laser Raman spectroscopy of nu 1, and the fluorescence of nu 1 labeled with N-(3-pyrene) maleimide on thiol groups of H3 histone were employed to detect conformational transitions in nu 1. The results of pH studies were as follows: 5.5 greater than pH greater than 4.8, suppression of DNA ellipticity and no change of histone alpha-helix; 4.6 greater than pH greater than 4.2  an irreversible increase in the B character of DNA, a slight loss of histone alpha-helix, and a parallel loss of pyrene excimer fluorescence; 4 greater than pH, aggregation of nu 1 and protonation of the DNA bases C and A. Results obtained in the studies of nu 1 in solvent mixtures included the following: sharp conformational transitions that variously involved an increase in the B character of DNA, a slight loss of histone alpha-helix, and a loss of pyrene excimer. Different solvents required different concentrations to effect these conformational changes.


Studies on proliferation kinetics in liver and spleen during early graft-vs-host reaction in mice. Autoradiographic and liquid scintillation counting studies were made during the early graft-versus-host-reaction in mice. At the height of GVHR on the 10th day after transplantation, hepatosplenomegaly correlated with the radioactive disintegrations (DPM). This coincided with a high proliferation rate of the blasts that appeared in the spleen, and of the portal infiltrations and Kupffer cells in the liver. Convenient control groups were used to show that these changes are a specific reaction in GVHR.


A case of multiple endocrine adenomatosis with primary amenorrhoea. A well documented sporadic case of multiple endocrine adenomatosis (MEA) type I, with the pituitary tumour presenting as a prolactinoma, is described in a 28-year-old female. Primary amenorrhoea, resulting from hyperprolactinaemia, was the first symptom of the polyglandular neoplasia. A gastrinoma was removed from the head of the pancreas and latent hyperparathyroidism appeared to be present. Treatment with bromocriptine was poorly tolerated; neurosurgical intervention was refused by the patient. The possibility that a serum prolactin determination may be useful in detecting pituitary involvement in MEA deserves consideration.


Endocrinological control of growth.

Endocrinological control of growth. Many endocrinological factors control cellular growth of different tissues (cell multiplication and cell volume) and skeletal growth. The role of neuro-transmitters and of hypothalamic releasing and inhibiting factors of growth hormone secretion will be reviewed. The importance of the somatomedins on cartilage growth will be stressed. Thyroid hormones, androgens, and oestrogens have important stimulating actions on skeletal growth and maturation. Conversely, glucocorticoids have an important inhibitory effect on growth. The precise roles of these hormone factors in the regulation of growth hormone secretion, somatomedin production and tissue growth, particularly the cartilage, remain to be completely elucidated.


[The role respiratory acidosis in the development of acute pancreatitis (author's transl)]. The effects of respiratory acidosis on the pancreas were studied in 25 Wistar rats. Ligation of the duct combined wiht respiratory acidosis resulted in the development of acute pancreatitis. In explantation of the results it is suggested that exhaustion of the buffering capacity causes a dramatic fall in pH  of the pancreatic juice. The clinical relevance of findings is discussed.


[Preparation of extracellular ribonuclease form Actinomyces rimosus 994]. By sequential acid treatment, gel filtration and KM-cellulose sorption a 18--20-fold purified preparation of ribonuclease with a yield of 50--60% was obtained from the culture liquid filtrate of Actinomyces rimosus 994. The preparation had a high specific activity of 450,000--600,000 units/mg protein, contained 85--98% protein, insignificant amounts of carbohydrates and hydroxytetracycline, and no quantities of DNase, phosphomonoesterases, phosphodiesterase or proteases. In RNA degradation (preparation of the total yeast RNA of the Sigma Co.) optimal results were obtained at 50 degrees C and pH 7.0--7.2 in phosphate buffer and 7.6--8.0 IN Tris-HCl buffer. The preparation was stable at high temperatures (80--100 degrees) in the wide pH range and during storage in the lyophilized form and in buffer solutions. RNase effect was inhibited by zinc, copper, iron and cobalt cations and activated by beta-mercaptoethanol, citrate and EDTA. Protamine sulphate and urea in low concentrations (0.01% and 1--4 M, respectively) accelerated and in high concentrations (1% and 8 M, respectively) terminated the enzyme reaction. With respect to many properties RNase from Act. rimosus 994 was similar to extracellular RNases, produced by other actinomycetes and fungi.


[Trypsin immobilization on a mineral matrix]. Trypsin immobilization on the mineral matrix--silochrome was studied. The effect of the matrix electrochemical nature on the process was examined. pH-optima for trypsin binding with different silochromes and pH-optimum of action of the immobilized enzyme on casein were determined. The effect on the trypsin-silochrome binding of different supplements--inhibitors (benzamidine), stabilizers (Ca2+) and substrates (casein, benzoyl argininamide hydrochloride) was demonstrated.


[Properties of immobilized acid proteinase from Aspergillus awamori]. pH-optimum, thermal stability and storage stability of immobilized acid proteinase from Aspergillus awamori obtained by covalent binding on silochrome through glutaraldehyde were studied. Acid proteinase immobilization was found to shift towards the acid range by a unit. Thermal stability of the immobilized preparation was lower than that of the native enzyme.


Stoichiometry of vectorial H+ movements coupled to electron transport and to ATP synthesis in mitochondria. In order to verify more directly our earlier measurements showing that, on the average, close to four vectorial H(+) are rejected per pair of electrons passing each of the three energy-conserving sites of the mitochondrial electron transport chain, direct tests of the H(+)/2e(-) ratio for sites 2 and 3 were carried out in the presence of permeant charge-compensating cations. Site 2 was examined by utilizing succinate as electron donor and ferricyanide as electron acceptor from mitochondrial cytochrome c; the directly measured H(+)/2e(-) ratio was close to 4. Energy-conserving site 3 was isolated for study with ferrocyanide or ascorbate plus tetramethylphenylenediamine as electron donors to cytochrome c and with oxygen as electron acceptor. The directly measured H(+)/2e(-) ratio for site 3 was close to 4. The H(+)/ATP ratio (number of vectorial H(+) ejected per ATP hydrolyzed) was determined with a new method in which the steady-state rates of both H(+) ejection and ATP hydrolysis were measured in the presence of K(+) + valinomycin. The H(+)/ATP ratio was found to approach 3.0. A proton cycle for oxidative phosphorylation is proposed, in which four electrochemical H(+) equivalents are ejected per pair of electrons passing each energy-conserving site; three of the H(+) equivalents pass inward to derive ATP synthesis from ADP and phosphate and the fourth H(+) is used to bring about the energy-requiring electrogenic expulsion of ATP(4-) in exchange for extramitochondrial ADP(3-), via the H(+)/H(2)PO(4) (-) symporter.


[Effect of carbon composition of the fermented medium on the synthesis of volatile acids by the yeast Saccharomyces carlsbergensis 776]. The effect of glucose, maltose and sucrose on the synthesis of volatile oils during fermentation of model carbohydrate solutions (6, 8 and 11%) by the yeast Saccharomyces carlsbergensis 776 was studied. The composition and concentration of carbohydrates affected the build-up of volatile fatty acids during fermentation. The accumulation of biomass and volatile fraction of fatty acids reached maximum on the medium containing 11% glucose. There was a certain correlation between the biomass synthesis and accumulation of volatile fatty acids, i.e. with an increase in the biomass the content of volatile fatty acids in the medium increased. During fermentation of disaccharide solutions cell multiplication diminished and the fermentation process accelerated. Ethanol, residual sugar and acidity of fermented solutions increased with the initial concentration of carbohydrates. The pH value of the fermented must remained essentially unaltered independent of the sugar amount used.


Evidence that the gamma-glutamyl cycle functions in vivo using intracellular glutathione: effects of amino acids and selective inhibition of enzymes. The function of the gamma-glutamyl cycle was explored in in vivo studies in which amino acids and specific inhibitors of cycle enzymes (gamma-glutamyl transpeptidase, gamma-glutamyl cyclotransferase, gamma-glutamylcysteine synthetase, and 5-oxoprolinase) were administered to mice. The findings, which show that the gamma-glutamyl cycle functions in vivo, support the conclusion that gamma-glutamyl amino acids formed by gamma-glutamyl transpeptidase from externally supplied amino acids and intracellular glutathione are translocated into the cell and thus indicate that there is a significant physiological connection between the metabolism of glutathione and the transport of amino acids.


Synthesis of some alkoxybenzamide derivatives as smooth muscle relaxant agents. The preparation of 16 new N-substituted p-aminobenzamide derivatives which contain diethylaminoethyl-N-methylpiperazine, piperidine and morpholine moieties were described. Preliminary pharmacological testing of representative compounds showed that some of the prepared compounds possess anticholinergic and antihistaminic activities.


Dehydration of 2-hydroxy(ethoxy)-2-aryl-N.N-penta(tetra) methylenemorpholinium bromides. Dehydration of 2-hydroxy(ethoxy)-2-aryl-N.N-penta(tetra) methylenemorpholinium bromide was achieved either by heating (175 degrees C) the compound with catalytic amounts of hydrobromic acid, or by refluxing the compound in a glacial acetic acid-hydrogen bromide mixture or by treating it with phosphorus tribromide at room temperature. In the acetic acid-hydrogenbromide method, the product was accompanied by N-bromoethyl(carbethoxy)-N-phenacylmorpholinium bromide, while strong heating (210 degrees C) of the title compounds or reflux in xylene yielded N-2-bromoethylpiperidine hydrobromide. Under the latter conditions, N-methyl-N-phenacylpiperidinium bromide gave similarly N-methylpiperidine hydrobromide. A preliminary biologic activity study in the synthesized compounds has also been conducted.


Aspects of application of cytochrome P-450 and related systems in substrate hydroxylation. Extrapolating the recent progress in the near future the extensive utilization of cofactor-dependent enzymes (enzymes of the 3rd generation) for solving economic or medical problems will be restricted by the difficulties of cofactor regeneration. Real possibilities exist in analytical systems, for instance enzyme electrodes. In the present paper a special case of overcoming the cofactor regeneration in P-450 catalyzed substrate hydroxylation is demonstrated: The peroxide-dependent reaction gives the same products as obtained under physiological conditions; that is why in an electro-enzyme-reactor producing hydrogen peroxide by cathodic oxygen reduction a considerable simplification of the multi-enzyme complex is possible by omitting electron transfer proteins. At present the main problem is the instability of the terminal oxidase. Attempts are being made to solve these problems by immobilizing the protein or substituting P-450 by other hemoproteins or iron porphyrin derivatives.


Polarographic estimation of antazoline hydrochloride. The dc-polarographic investigation of antazoline hydrochloride in aqueous acidic media is described. Using potassium chloride-hydrochloric acid mixture as the supporting electrolyte (pH = 3.25), antazoline hydrochloride was electrochemically reduced at the dropping mercury electrode, with the production of two waves with E1/2 values of --1.35 and --1.65 V respectively. As revealed from the study of the effect of mercury column height, pH of the medium and concentration of the depolarizer, the polarographic reduction of the antazolinium cation is preceded by a catalytic H-wave. The diffusion-controlled nature of the electrode process permitted the quantitative determination of antazoline hydrochloride in concentrations down to 1.0 . 10(-4)M. Application of the presented procedure to the analysis of different dosage forms of the compound studied proved successful and compared favourably with official estimations of anatazoline salts. In view of its simplicity, accuracy and sensitivity, the presented polarographic method can be recommended for routine analysis of antazoline formulations.


A model of the pH-dependence of the number of oxygen-linked chloride binding sites in hemoglobin. A model is presented of the pH-dependence of the number of oxygen-linked chloride binding sites established by nuclear magnetic resonance quadrupole-relaxation studies on various mutant and chemically modified hemoglobins. The predictions of the model are in good qualitative agreement with the measured pH-dependences of the linewidth of the 35Cl- NMR signal. The obtained agreement implies that more chloride is bound to oxygenated than to deoxygenated hemoglobin.


Stability of horse muscle acylphosphatase to heat and to urea. The thermal stability of horse muscle acylphosphatase was investigated by measuring the inactivation constants at various pH and temperature values, and by differential spectra technique. This enzyme has high thermal stability in an acidic environment but is inactivated in an alkaline medium. It was found that the enzyme can be protected against such inactivation at pH 8.0 by increasing its concentration and the ionic strength of the solution. The effect of high urea concentrations on stability was also measured. It was found that spectral changes at 230 nm are related to urea inactivation of the enzyme, and that the enzymatic activity can be instantly and almost completely restored by dilution of the urea.