Physiology and chemistry of cerebrospinal fluid, aqueous humor and endolymph in Squalus acanthias. By means of the appropriate isotopes injected into the spiny dogfish, Squalus acanthias, the transfer of all major ions into cerebrospinal fluid (CSF), aqueous humor (A) and endolymph (E) was studied. In addition, the effect of raising pCO2 in sea-water upon HCO3- concentration of these fluids was measured. In the several types of experiments, acetazolamide or methazolamide was used to inhibit completely carbonic anhydrase. The rates of fluid formation and ion transfer in CSF and A were fairly close, but those for E were far slower. The general pattern of ion transport in the three fluids were the same, Na+ (or Na+ + K+ in E) entry greater than Cl - entry, and the difference was HCO3-. The greater rate constants for HCO3-, increase in its entry rate by elevation of pCO2, and inhibition of its appearance by the sulfonamides, show that this is a special case  of transport; the ion is formed in secretory cells from gaseous CO2 + OH-. Secretory cells at sites of formation of all the fluids contain both carbonic anhydrase and Na+-K+-ATP-ase, which subserve HCO3- formation and Na+ (or K+) transport. Comparison of these results with studies in mammals show that the vertebrate pattern for secretion of these three fluids is well established in the elasmobranch.


Characterization of gill (Na + K)-activated adenosine triphosphatase from chinook salmon, Oncorhynchus tshawytscha. (Na+K)-activated ATPase activity from gills of yearling spring chinook was examined using a new rapid assay method. Characterization of the enzyme activity was performed. Optimal activity was obtained at pH 7.2 in the presence of 240 mM NaCl, 120 mM KCl, 20 mM MgCl2 and 10 mM Na2ATP. Maximal inhibition of the enzyme was observed in the presence of 0.5 mM ouabain. Differential centrifugation indicated that 75% of the enzymatic activity was sedimented at 1000 x g. Only 8% of the activity was found in the microsomal pellet. Treatment with 0.1% sodium deoxycholate liberated activity from the 1000 x g pellet and elevated the activity. This treatment caused a loss of 20% of the original activity of the preparation. Statistical analysis of the sampling procedure for gill (Na+K)-activated ATPase activity indicated that there was small variation in the technique itself when compared to variation between the individual gill arches and between individual fish. Results indicate that for meaningful comparisons of groups of fish, the sampling of the gill arches must be standardized and a large number of individual fish must be sampled.


Correlation of structure and active transport in the teleost nephron. In the present study we have extended our investigations concerning the correlation between ultrastructure and active transport in the isolated flounder nephron. The composition of the fish nephron is defined in ultrastructural terms and its behavior when incubated in vitro under short term and long term culture conditions is described. Using the in vitro system originally described by Forster, a variety of inhibitors and conditions which modify cell structure and function were tested. Ultrastructure was correlated with chlorphenol red dye transport. In general, conditions altering active transport also markedly altered cellular ultrastructure. The principal alterations consisted of membrane changes involving various organelles--most importantly the plasma membrane and the mitochondria. Conditions associated with irreversible cell injury could be rapidly produced by interference either with mitochondrial ATP synthesis or with the integrity of the plasma membrane. Both of these rapidly lead to irreversible events which are preceded by reversible structural changes. Organelle changes progress in a rather well-defined sequence of reversible and irreversible stages which are defined. One difference between the two types of interactions is the presence of intramitochondrial calcification which does not occur with direct modification of the mitochondrial electron transport system. The concept of utilizing long term explant organ cultures of fish nephrons for environmental studies is introduced.


Characteristics of the chloride conductance in muscle fibers of the rat diaphragm. In muscle fibers from the rat diaphragm, 85% of the resting membrane ion conductance is attributable to Cl-. At 37 degree C and pH 7.0, GCl averages 2.11 mmho/cm2 while residual conductance largely due to K+ averages 0.34 mmho/cm2. The resting GCl exhibits a biphasic temperature dependence with a Q10 of 1.6 between 6 degree C and 25 degree C and a Q10 of nearly 1 between 25 degree C and 40 degree C. Decreasing external pH reversibly reduced GCl; the apparent pK for groups mediating this decrease is 5.5. Increasing pH up to 10.0 had no effect on GCl. Anion conductance sequence and permeability sequence were both determined to be Cl-greater than Br-greater than or equal to I-greater than CH3SO4-. Lowering the pH below 5.5 reduced the magnitude of the measured conductance to all anions but did not alter the conductance sequence. The permeability sequence was likewise unchanged at low pH. Experiments with varying molar ratios of Cl- and I- indicated a marked interaction between these ions in their transmembrane movement. Similar but less striking interaction was seen between Cl- and Br-. Current-voltage relationships for GCl measured at early time-points in the presence of Rb+ were linear, but showed marked rectification with longer hyperpolarizing pulses (greater than 50ms) due to a slow time-and voltage-dependent change in membrane conductance to Cl-. This nonlinear behavior appeared to depend on the concentration of Cl- present but cannot be attributed to tubular ion accumulation. Tubular disruption with glycerol lowers apparent GCl but not GK, suggesting that the transverse tubule (T-tubule) system is permeable to Cl- in this species. Quantitative estimates indicate that up to 80% of GCl may be associated with the T tubules.


The relationship between anion exchange and net anion flow across the human red blood cell membrane. The conductive (net) anion permeability of human red blood cells was determined from net KCl or K2SO4 effluxes into low K+ media at high valinomycin concentrations, conditions under which the salt efflux is limited primarily by the net anion permeability. Disulfonic stilbenes, inhibitors of anion exchange, also inhibited KCl or K2SO4 efflux under these conditions, but were less effective at lower valinomycin concentrations where K+ permeability is the primary limiting factor. Various concentrations of 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS) had similar inhibitory effects on net and exchange sulfate fluxes, both of which were almost completely DIDS sensitive. In the case of Cl-, a high correlation was also found between inhibition of net and exchange fluxes, but in this case about 35% of the net flux was insensitive to DIDS. The net and exchange transport processes differed strikingly in their anion selectivity. Net chloride permeability was only four times as high as net sulfate permeability, whereas chloride exchange is over 10,000 times faster than sulfate exchange. Net OH-permeability, determined by an analogous method, was over four orders of magnitude larger than that of Cl-, but was also sensitive to DIDS. These data and others are discussed in terms of the possibility that a common element may be involved in both net and exchange anion transport.


Extrasynaptic receptors on cell bodies of neurons in central nervous system of the leech. 1. A systematic study has been made of the sensitivity of identified sensory and motoneurons in the leech central nervous system to chemical transmitter substances. 2. The following substances elicited responses from the cell bodies of individual neurons: acetylcholine, 5-hydroxytryptamine, gamma-aminobutyric acid, glutamic acid, glycine, dopamine, and norepinephrine. Since the cell bodies of leech neurons are free of synapses, the receptors that give rise to these responses are extrasynaptic. 3. Sensory and motoneurons of different function had characteristic complements of extrasynaptic receptors. For example, mechanosensory cells responding to light touch, to pressure, and to noxious stimuli could be distinguished by their responses to iontophoretically applied compounds. For one of these modalities (nociceptive), neurons with different receptive fields but otherwise similar properties had markedly distinct extrasynaptic receptors. The possible significance of extrasynaptic receptors is discussed.


Synthesis of acetylcholine by excitatory motoneurons in central nervous system of the leech. 1. A study was made of the synthesis of acetylcholine (ACh) and other transmitters by the cell bodies of functionally identified neurons in leech segmental ganglia. 2. Choline acetyltransferase, the synthetic enzyme for ACh, was detected in excitatory motoneurons but not in mechanosensory cells or Retzius cells. The ability of motoneurons to synthesize ACh was also demonstrated by their accumulation of [3H]ACh following incubation of segmental ganglia with [3H]choline. [3H]ACh was not detected in the other cell types. When eserine was included in [3H]choline incubations, the amount of [3H]ACh in motoneurons increased severalfold and small amounts of [3H]ACh (1% that in motor cells) appeared in extracts of sensory and Retzius cells. 3. In addition to [3H]ACh segmental ganglia synthesized [3H]5-HT, [3H]gamma-aminobutyric acid, [3H]dopamine, and [3H]octopamine from exogenous, labeled precursors. None of these labeled transmitters was detected in identified neurons except [3H]5-HT, which was found in Retzius cells. 4. These results provide biochemical evidence that excitatory motoneurons in the leech are cholinergic, but leave open the identity of the sensory transmitter(s).


Filterability of erythrocytes from vitamin E-deficient lead-poisoned rats. The time required for red blood cells (RBC) from vitamin E-deficient lead-poisoned (-E + Pb) rats to pass through polycarbonate filters after incubation in vitro was much greater than that of RBC from vitamin E-supplemented non-poisoned rats. Vitamin E deficiency per se (i.e., in non-poisoned rats) often increased filtration times, but in all such experiments the RBC from -E + Pb groups had even longer filtration times. Administration of lead to rats supplemented with vitamin E had little effect on the filtration rate of RBC. N,N'-diphenyl-p-phenylenediamine (DPPD) prevented the increased filtration times characteristic of RBC from -E + Pb rats, but replacement of the lard in the vitamin E-deficient basal diet by more highly polyunsaturated fats did not exacerbate the increased filtration times of RBC from -E + Pb rats. The increased filtration time of RBC from -E + Pb rats appeared to be related to the extent of RBC lipid peroxidation. Decreasing the pH of the RBC incubation medium from 7.4 to 6.6, an acidity typical of the spleen, markedly increased the filtration times of RBC from -E + Pb rats. Addition of lead in vitro increased filtration times of RBC from both vitamin E-deficient and supplemented non-poisoned rats, but filtration times tended to be longer in the deficient group. These results suggest that vitamin E deficiency and lead toxicity act synergistically to alter the deformability of the RBC thereby rendering it vulnerable to sequestration in the spleen.


Utilization of riboflavin homologues by D-amino acid oxidase and xanthine oxidase. D-Amino acid oxidase and xanthine oxidase, two enzymes possessing ionically bound flavin coenzymes have been studied with their flavin coenzymes derived from either 7-ethyl-8-methyl-flavin or 7-methyl-8-ethyl-flavin, vitamin-like homologues of riboflavin. 7-Ethyl-8-methyl-flavin caused a significant reduction of both D-amino acid oxidase and xanthine oxidase in the liver, but not in the kidney. 7-Methyl-8-ethyl-flavin caused a significant reduction of D-amino acid oxidase in both the liver and kidney, a significant reduction of xanthine oxidase in the liver, but a large and significant increase of the latter enzyme in the kidney. An improved procedure for the assay of xanthine oxidase has been described.


Beneficial effect of methionine and threonine supplements on tyrosine toxicity in rats. The growth retardation and external pathological lesions that occur in rats fed a 10% casein diet containing 5% tyrosine could be alleviated by the supplementation of extra protein (20% casein) or 0.66% methionine plus 0.90% threonine (equivalent to the contents in a 20% casein, respectively). Liver tyrosine aminotransferase activity is elevated by ingestion of excess tyrosine, but lowered by the supplementation of extra casein of methionine plus threonine. In rats fed a high tyrosine diet supplemented with methionine plus threonine, liver p-hydroxyphenylpyruvate hydroxylase activity was not higher than that of the 10% casein group, but liver homogentisate oxidase activity increased significantly. When excess tyrosine was included in the 10% casein diet, free tyrosine concentrations in plasma, liver, muscle and brain were extremely elevated, but when the high tyrosine diet was supplemented with extra casein or methionine plus threonine, their plasma and tissues tyrosine concentrations lowered significantly. A large increase in total phenols, p-hydroxyphenylpyruvate and free tyrosine excretions in urine was produced in animals fed the high tyrosine-low protein diet, but these were lowered by the supplementation of methionine and threonine to the diet.


Blood and brain concentrations of imipramine, clomipramine and their monomethylated metabolites after oral and intramuscular administration in rats. Imipramine and clomipramine were administered to rats by the oral and intramuscular routes as single and multiple doses. The concentrations of both drugs and their active demethylated metabolites desipramine and desmethylclomipramine were measured in blood plasma, blood cells and brain. The concentrations of the metabolites were higher and the concentrations of the parent substances lower after oral than after parenteral administration, both in blood and in brain. In brain imipramine, despiramine and clomipramine during continuous treatment exceeded their plasma concentrations by six to ten times. The corresponding figure for desmethylclomipramine was 1-7. The extent of accumulation of the investigated substances in the brain was independent of the route of administration.


Papaver bracteatum Lindley: thebaine content in relation to plant development. Four thebaine-rich varieties of P. bracteatum have been grown in the open over two seasons and the thebaine distribution in aerial parts examined to determine the most suitable source material for commercial production. The leaves contained only 0-1 to 0-15%; the capsules 0-5 to 3-0% and the bled latex 28 to 53%. The maximum for the latter occurred about 3-4 weeks after petal opening and during the day, at about 15,00 h. A product 'bractium' prepared exactly as opium from P. somniferum contained up to 55% thebaine and calculations from the 1974 results gave theoretical yields up to 58 k of thebaine per hectare. However this is a very labour intensive method; furthermore bled latex only represents about 46% of the total thebaine of the capsule. In addition the pedicels contain significant amounts of thebaine, so that fruiting tops may be recommended as source material. In the capsule the thebaine content reaches a peak 3 to 4 weeks after petal opening and again two weeks later. At this fully ripe stage there is a theoretical yield of 50 kg per hectare. Two further advantages accrue from collection at this time: the ripe seeds can probably be used for similar purposes as poppy seed; and the pericarps at this stage contain no 'bound thebaine' (i;e., thebaine insoluble in MeOH; NH4OH but soluble in acetic acid--in unripe capsules bound thebaine represents 18 to 36% of the total thebaine). There is some evidence that, as this perennial plant increases in age, the capacity for thebaine production seems to continue increasing. Storage of raw material, even in ideal conditions, led to a loss of thebaine of 12 to 20% in one year.


Dimensional changes of compacts after compression. A non-contact optical technique has been used to measure changes in the heights and diameters of compacts prepared from sodium chloride, spray dried lactose, two samples of methylcellulose powder and two spray-dried lactose-maize starch granulations. Both sodium chloride and spray dried lactose exhibited relatively small dimensional changes whilst the methylcellulose powders showed up to 30% axial and 3% radial expansion. The results are discussed in terms of the inherent properties of the materials. When the lactose was granulated with maize starch, the ratio of axial; radial strain recovery was reduced from 5-9:1 to approximately 1:1, suggesting an improved distribution of forces during compression of the granulations,


Moisture and gelatin effects on the interparticle attractive forces and the compression behaviour of oxytetracycline formulations. The tensile strengths of compacts and/or tablets of the individual components and of granules prepared from an oxytetracycline formulation have been measured using the diametral compression test. Employing the theory of tensile strength, proposed by Cheng, it has been shown that increases in both moisture and gelatin contents of compacts and tablets increase the range of the attractive forces that operate between the granules. By studying the effects of moisture and gelatin on the compressional behaviour of the granules, it has been possible to classify them into different types. Fragmentation of granules occurs at packing fractions between 0-745 and 0-835, depending on their gelatin content.


The properties of tablets containing microcrystalline cellulose. Tablets have been prepared from mixtures of microcrystalline cellulose (Avicel) and spray dried lactose. Tests on these showed that a maximum value of dissolution rate occurred as the percentage of the cellulose increased. This maximum was at 4% w/w for mixtures containing Avicel PH 101 and coincided with the point of maximum liquid penetration rate. With grade PH 105 this maximum was at 2% w/w and corresponded to the optimum balance between the opposing factors of disintegration and liquid penetration rate. Addition of up to 2% w/w magnesium stearate to the formulation containing 4% w/w PH 101 grade had little effect on pore structure, but decreased the dissolution rate by retarding water penetration. Similar concentrations of Carbowax 4000 caused no such decrease.


The influence of temperature on the sorption of benzocaine by nylon 6 from aqueous cosolvents. The influence of temperature on the sorption of benzocaine by nylon 6 powder from aqueous cosolvents has been assessed. At about 30 degrees discontinuities appear in the Van't Hoff plots for sorption from water and aqueous PEG 400, which may be attributed to transitional changes in the polymer structure. No such discontinuities are apparent for sorption from aqueous ethanol over the temperature range 15-60 degrees, which is indicative of plasticizaton by this cosolvent.


The identification and quantitation of the major metabolites of ethylamphetamine, produced by rabbit liver microsomal preparations. Amphetamine (I), 1-phenylpropan-2-ol (III), phenylacetone (V), N-hydroxyethylamphetamine (VII), and alpha-methyl-N-(1'-phenylprop-2'-yl)nitrone (VIII) were obtained as metabolic products from the incubation of ethylamphetamine (II) with fortified male rabbit liver 9000 g supernatant fractions. These metabolites were identified by comparing their t.l.c., g,l.c. and g.l.c.-m.s. behaviour to that of reference compounds. A method for the quantitative analysis of the metabolites in mixtures using g.l.c. techniques is described.


Effect of oxotremorine and sodium pentobarbitone on the pharmacokinetics of intravenous tracer doses or radioactive choline. The present study explains for the previous findings that oxotremorine increases and sodium pentobarbitone decreases the initial brain uptake of intravenously injected radioactive choline (3H-Ch). The effects are explained by haemodynamic changes since a corresponding increase and decrease in the plasma concentrations of 3H-Ch were found. This is important to know when 3H-Ch is used for estimation of aceytlcholine turnover in the brain. The drugs did not affect the distribution of radioactivity (3H) between plasma and erythrocytes.


The fate of prostaglandin A1-5,6-3H in the rat. The plasma concentrations, tissue distribution and excretion of prostaglandin A1 (PGA1) and related metabolites have been determined in rats, following the intravenous injection of a single dose of PGA1-5,6-3H. Urinary and faecal excretion accounted for averages of 25 and 43% of the administered dose of PGA1, respectively. Oxidative cleavage of the carboxyl side chain of PGA1 appeared to be a major metabolic pathway in the rat. PGA1-5,6-3H was deemed unsuitable for metabolism studies in man, in view of the significant loss of tritium label from the prostaglandin;


Possible subdivisions among alpha-adrenoreceptors in various isolated tissues. The ratio (expressed in log10 units) of the equieffective concentrations of (+)- and (-)-noradrenaline has been measured in a variety of isolated tissues in the presence of cocaine (1 x 10(-5) M), tropolone (3 x 10(-5) M) and (+/-)-propranolol (5 x 10(-7) to 5 x 10(-5) M). The values obtained fall into 3 distinct and statistically different groups. Firstly, a high group comprising (mean +/- s.e.) mouse vasdeferens (2-78 +/- 0-04), rabbit duodenum (2-91 +/- 0-07) and ileum (2-86 +/- 0-05). Secondly a middle group comprising rabbit vas deferens (2-54 +/- 0-04), bladder neck muscle (2-56 +/- 0-07) and spleen 2-50 +/- 0-02), guinea-pig vas deferens (2-55 +/- 0-10) and bladder neck muscle (2-48 +/- 0-13) and rat deferens (2-40 +/- 0-08) and thirdlya low group comprising the bladder detrusor muscle from both the rabbit (2-08 +/- 0-08) and the guinea-pig (2-07 +/- 0-04). Under the same conditions measurement of pA2 values for phentolamine and piperoxan against noradrenaline gave the following values in rat vase deferens (8-22 +/- 0-07 and 6-72 +/- 0-03 respectively) and mouse vas deferens (8-31 +/- 0-05 and 6-53 +/- 0-07 respectively). The results are discussed in relation to other findings conderning the nature of the alpha-adrenoreceptor in these tissues. In spite of the absence of any significant difference between the potency of the alpha-adrenoreceptor blocking agents in the two species it is suggested that alpha-adernoreceptors may not belong to a single homogenous population but may vary in their characteristics from tissue to tissue.


Effect of butaclamol, a new neuroleptic, on serotoninergic mechanisms. Butaclamol (1.0-0.1 mg kg-1, i.p.) and spiroperidol (1-0-0-5 mg kg-1, i.p.) but not (-)-butaclamol (15 mg kg-1, i.p.), blocked the hyperactivity induced in rats by tranylcypromine-L-tryptophan pretreatment. Neither butaclamol nor spiroperidol altered the accumulation of brain 5-HT following parglyine or the decline of brain 5-HT following inhibition with the tryptophan hydroxylase inhibitor alpha-propyldopacetamide thus indicating that butaclamol and spiroperidol do not affect either the synthesis or the turnover of brain 5-HT. It is concluded that the antagonism of the tranylcypromise-L-tryptophan-induced hyperactivity by butaclamol and spiroperidol is due to their blockade of dopaminergic receptors rather than an action on neuronal serotoninergic mechanisms.


Effect of lofepramine and other antidepressants on the uptake of 5-hydroxytryptamine and noradrenaline into rat brain monoaminergic neurons. Lofepramine, (N-methyl-N-[4-chlorobenzoylmethyl]-3-[10,11-dihydro-5H-dibenz(b,f)-azepin-5-yl]-propylamine hydrochloride), is a new antidepressant with low toxicity and no peripheral anticholinergic activity. Its effect on 5-hydroxytryptamine (5-HT) and noradrenaline uptake into rat brain monoaminergic neurons was studied and compared with that of other antidepressants, particularly with that of imipramine and desipramine. Lofepramine inhibited both 5-HT and noradrenaline uptake into synaptosomal fractions in vitro but was 4 times more potent in inhibiting noradrenaline than 5-HT uptake, indicating the effect resembles that of desipramine. Noradrenaline uptake was also preferentially inhibited in synaptosomes from brain of rats treated previously with lofepramine or desipramine (i.p.). Pretreatment with SKF 525A (i.p.) did not diminish the effect of lofepramine, but rather potentiated it. Therefore it is suggested that the formation of desipramine is not necessary for lofepramine to exhibit, the effect on amine uptake in vivo. Both lofepramine and desipramine inhibited intraventricular noradrenaline uptake into synaptosomes without any effect on 5-HT uptake. These results suggest that lofepramine is qualitatively similar to desipramine with respect to preferential inhibition of noradrenaline uptake into central noradrenergic neurons.


Hyperpyrexic interaction between debrisoquine and pethidine in rabbits. Pethidine injection into rabbits treated with debrisoquine either acutely or chronically resulted in severe interaction and fatal hyperpyrexia. Pretreatment of rabbits with p-chlorophenylalanine, chlorpromazine, or crypoheptadine protected them against the interaction, while alpha-methyl-p-tyrosine was ineffective. In addition the administration of debrisoquine into 5-HTP pretreated rabbits produced a severe interaction and hyperpyrexia. The hepatic N-demethylation of pethidine was significantly inhibited by debrisoquine pretreatment both in vivo and in vitro. The debrisoquine-pethidine interaction could be due to 5-HT potentiation or prevention of uptake. Alternatively it could be due to inhibition of biotransformation of pethidine by debrisoquine. However, neither mechanism by itself alone could be held responsible as the sole explanation of the interaction.


Synthesis and pharmacological properties of a series of antidopaminergic piperidyl benzamides. The synthesis and pharmacological screening for anti-apomorphine, stomach emptying and local anaesthetic activities of some new piperidylbenzamides is described. One of these, N-(1'-benzyl-4'-piperidyl)-2-methoxy-4-amino-5-chlorobenzamide (clebopride) is more potent than metoclopramide in tests related to blockade of cerebral dopamine receptors.


Conformational analysis of dopamine by the INDO molecular orbital method. The results of INDO calculations on dopamine are reported. A conformational energy map and an isodistance map for the key distances N-OH1, N-OH2 in dopamine as functions of the two main torsion angles tau1 and tau2 were constructed. In addition to the three known minima of dopamine corresponding to the trans and gauche forms, two new minima were found. The key distances of the rigid analogues of dopamine, apomorphine, isoapomorphine, 2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene and isoquinoline were plotted on the isodistance map of dopamine. By taking the corresponding tau values as coordinates on the energy map, conformations of dopamine, resembling the rigid analogues, could be found. When a conformation is close to a local minimum it is assumed that this conformation is energetically favourable. The possible relation between the energy minima and the biological action of dopamine is discussed. An explanation is suggested for the lack of dopaminergic activity of isoapomorphine.


Structure and rheology of cetomacrogol creams: the influence of alcohol chain length and homologue composition. Liquid paraffin-in-water emulsions prepared with cetomacrogol 1000 and alcohols cetostearyl (A), cetyl (B), steryl (C) and myristyl (D) were examined by microscopical, particle size analytical and rheological (continuous shear, small strain creep, and oscillation) techniques at 25 degrees as they aged over 30 days. The particle sizes of the emulsions were similar and did not increase significantly with age. Thus the rheological stabilities were not correlated with particle size distributors, but rather with viscoelastic networks formed in the continuous phases when the non-ionic mixed emulsifiers interacted with water. The rheological properties of emulsions B and D differed from those of emulsion C. Emulsion A, of mixed homologue composition, showed some properties similar to each of the pure alcohol emulsions. Emulsions B and D were semi-solid immediately after preparation whereas emulsion C was so mobile initially that small strain data were not derived. On ageing, the consistencies of B and D changed slightly initially, and then remained essentially constant. In contrast, the consistency of emulsion C increased on ageing, especially over the first few days when there was a change from mobile liquid to semisolid. Emulsion A was a semisolid initially but like emulsion C increased in consistency especially over the first 24 h. Continuous shear data indicated that this emulsion was the most resistant to structure breakdown. Microscopical examination supported the view that the networks formed in emulsion A were the most extensive and that stearyl alcohol networks in C formed comparatively slowly. Although the cetomacrogol/pure alcohol networks were diffuse and sometimes crystallized, they did not rapidly disintegrate on storage as did the ionic surfactant/pure alcohol networks examined previously.


Pharmaceutical suspensions: micro electrophoretic properties. The microelectrophoretic properties of the drugs griseofulvin, betamethasone, nalidixic acid and thiabendazole in aqueous dispersion have been examined and the zeta potentials calculated from the measured mobilities. Variation in magnitude of particle charge with pH of dispersion is reported and related to the chemical structure and the surface characteristics. The effect of adding anionic (sodium dodecyl sulphate) cationic (dodecyl trimethyl bromide) and non-ionic (polyoxyethylene glycol monoethers of hexadecanol) surface-active agents, and mixtures of these ionic and non-ionic species, on the electrophoretic properties of the drug dispersions has been measured. The results reported agree with those found previously for a model polystyrene latex suspension system under the same conditions.


Dissolution from tablets prepared using ethyl cellulose microcapsules. Microcapsules containing sodium phenobartitone cores in ethyl cellulose have been used to prepare tablets at from 3-9 to 358-9 MPa compression pressures. The tensile strength of these tablets is related linearly to the core: wall ratio and to the microcapsule size. Dissolution of the drug from the microcapsules, is also related to the core:wall ratio and microcapsule size, but except at low compression pressures is almost independent of the pressure used during preparation. The tablet matrix remains intact during the dissolution and the equations developed by Schwartz, Simonelli & Higuchi (1968) are followed. Large microcapsules 1:2 core: wall ratio produce friable tablets with rapid release of contents.


Cognitive processes mediating behavioral change. The present experiment was designed to test the theory that psychological procedures achieve changes in behavior by altering the level and strength of self-efficacy. In this formulation, perceived self-efficacy. In this formulation, perceived self-efficacy influences level of performance by enhancing intensity and persistence of effort. Adult phobics were administered treatments based upon either performance mastery experiences, vicarious experiences., or they received no treatment. Their efficacy expectations and approach behavior toward threats differing on a similarity dimension were measured before and after treatment. In accord with our prediction, the mastery-based treatment produced higher, stronger, and more generalized expectations of personal efficacy than did the treatment relying solely upon vicarious experiences. Results of a microanalysis further confirm the hypothesized relationship between self-efficacy and behavioral change. Self-efficacy was a uniformly accurate predictor of performance on tasks of varying difficulty with different threats regardless of whether the changes in self-efficacy were produced through enactive mastery or by vicarious experience alone.


Oxidative reactions of hydroxylated chlorpromazine metabolites. The oxidation pathways of two hydroxylated chlorpromazine metabolites were investigated using modern electrochemical techniques. Upon oxidation, the 7-hydroxy derivative of chlorpromazine rapidly reacts to form the 7,8-dihydroxy derivative and a substituted quinone. The oxidation potentials for both compounds were determined in the pH 3-8 range. The importance of these redox reactions and potentials to the pharmacology of the materials is discussed.


Electrochemical analysis of the cephalosporin cefamandole nafate. The polarographic assays for cefamandole sodium and its formyl ester, cefamandole nafate, are described. Controlled potential coulometry is used as an absolute method for the assignment of purity of these compounds without the need for a reference material. The precision, accuracy, and selectivity of these assays were better than for the microbiological autoturbidimetric and automated iodometric assays. NMR, TLC, GC, and polarography are used to detect and quantitate likely impurities and degradation products.


Tyrosine hydroxylase: delayed activation in central noradrenergic neurons and induction in adrenal medulla elicited by stimulation of central cholinergic receptors. The centrally active muscarinic agonist, oxotremorine, elicited an up to 2-fold dose-dependent (0.25-1.5 mg/kg) increase in the activity of tyrosine hydroxylase (TH) in the rat nucleus locus coeruleus (LC) and adrenal medulla. The response occurred in LC after 24 to 48 hours and in adrenal medulla by 4 to 8 hours, peaked in LC at 72 hours and adrenal medulla at 16 to 24 hours and persisted up to 2 weeks in both tissues. In brain the effect appeared confined to cell bodies of noradrenergic neurons. The activity of dopamine beta-hydroxylase increased in adrenal medulla (40%) but not in brain. Immunotitration with anti-TH serum demonstrated that the increase of TH activity in LC is due to increased catalytic activity (activation), whereas in adrenal medulla it is due to a transynaptically mediated accumulation of enzyme protein (induction). Physostigmine (1.0 mg/kg), pilocarpine (25-50 mg/kg) and nicotine (10 mg/kg) increased TH activity in LC and adrenal. We conclude that stimulation of central cholinergic receptors of the muscarinic type results in a delayed and protracted activaiton of TH but not of dopamine beta-hydroxylase in cell bodies of central noradrenergic neurons, and reflexly, to transynaptic induction of TH and dopamine beta-hydroxylase in the adrenal medulla.


Regulation of catecholamine biosynthesis in a transplantable rat pheochromocytoma. Cells prepared from a transplantable rat pheochromocytoma synthesize norepinephrine from 14C-tyrosine, at a rate of 9.4 +/- 0.5 pml/min/mg of protein, in vitro. Incubation of the cells in a medium containing 56 mM K+ results in a 2- to 6-fold increase in norepinephrine synthesis. This increase in norepinephrine synthesis is dependent upon the presence of Ca++ in the incubation medium. Stimulation of the cells by 56 mM K+ increases the conversion of tyrosine to dopa in the presence of brocresine (an inhibitor of aromatic L-amino acid decarboxylase), and has no effect on the conversion of 3H-dopa to norepinephrine. Cells can be depleted of up to 70% of their catecholamine stores by prior incubation in 56 mM K+. Norepinephrine synthesis in catecholamine-depleted cells incubated under control conditions in only slightly (20-40%) greater than it is in nondepleted cells. However, 56 mM K+ PRODUCES A SIMILAR INCREASE IN NOREPINEPHRINE SYNTHESIS IN DEPLETED CELLS AS IT DOES IN NONDEPLETED CELLS. Inhibition of amine oxidase (flavin containing) by preincubaiton with pargyline does not greatly affect catecholamine synthesis. Incubation of the cells in 56 mMK+ results in an increase in tyrosine 3-monooxygenase activity. These results indicate that the depletion of catecholamine stores plays only a minor role in the increase in norepinephrine synthesis caused by the stimulation of chromaffin cells and suggest that the activation of tyrosine 3-monooxygenase plays a more important role in this phenomenon.


Differential inhibition of lipolysis in human adipose tissue by adrenergic beta receptor blocking drugs. The effects of various adrenergic beta receptor agonists and antagonists on lipolysis (measured as glycerol release) in human adipose tissue in vitro were studied. Of the agonists investigated, the potency rank order was isoproteronol greater than norepinephrine greater than salbutamol. Adrenergic beta receptor blocking drugs inhibited catecholamineinduced lipolysis  competitively. Propranolol was the overall most effective compound, followed by metoprolol, alprenolol and practolol, whereas butoxamine and H35/25 were weak inhibitors. The results indicate that the adrenergic reciptor mediating lipolysis in human adipose tissue is of type beta-1. Basal and theophylline-induced lipolysis was reduced when higher concentrations of these drug were used.


Alterations in brain cyclic guanosine 3':5'-monophosphate levels after acute and chronic treatment with ethanol. A previous finding from our laboratory, that a single dose of ethanol depletes cerebellar cyclic guanosine 3':5'-monophosphate (cGMP), has now been extended to an investigation of the effects of acute and chronic ethanol treatment of cGMP levels in six areas of the rat brain. Rats were either gavaged with a single dose of ethanol (6g/kg) or rendered ethanol-dependent with 11 to 15 g/kg/day in 3 to 5 fractions over a 4-day period. A single dose of ethanol depleted cGMP levels in only five areas of the brain studied. In the two areas in which a time course of the response was determined, the caudate nucleus and cerebral cortex, cGMP depletion was maximal 1 hour after ethanol administration when blood ethanol concentrations were highest. cGMP levels returned to control values as blood ethanol was eliminated. Inethanol-dependent animals still intoxicated, cGMP was reduced but not to the same magnitude in the cerebellum and brain stem when compared with the response obtained after a single dose at equivalent blood ethanol concentrations. During the ethanol withdrawal syndrome cGMP levels had returned to control vlaues. The data suggest that cBMP depletion may play a role in ethanol-induced intoxication and that tolerance to this effect develops concurrently with behavioral tolerance.


Effect of palytoxin on membrane and potential and current of frog myelinated fibers. Palytoxin is a highly toxic compound isolated form several zoanthid Palythoa species. The effects of palytoxin on the nodal membrane of frog myelinated fiber have been studied under current clamp and under voltage clamp conditions. Under current clamp conditions, palytoxin (0.1 microng/ml, 3 x 10(-8)M) induces a depolarization which is not reversed by washing. The resting potential reaches a value of -35 mV after 10 minutes. During the same period, the evoked action potential shows a gradual decline and finally disapears after about 30 minutes. The membrane depolarization is suppressed by removal of Na ions from the external medium, but only slightly diminished when tetrodotoxin (10(-6)M) is subsequently added to the external medium. When the potential of the nodal membrane is maintained at -70 mV, palytoxin (0.1 microng/ml) causes the appearance of an inward current that increases in magnitude during 30 minutes before attaining a steady-state value. The kinetics of development of that current is modified in the presence of tetrodotoxin or saxitoxin. Voltage clamp analysis shows that palytoxin causes an increase of the resting sodium permeability that is accompanied by a shift of the voltage dependence of the transient sodium permeability in the direction of membrane hyperpolarization. The shift in the voltage dependence of the transient permeability is accompanied by a decrease of the peak transient permeability. A similar shift in the potential dependence of the sodium inactivation is observed. During and after the application of palytoxin, the internal sodium concentration increases. The steady-state (potassium) conductance is also decreased at the same time as the leak current is increasing.


Active form of ketamine in squid giant axons. The active form of ketamine has been studied with internally perfused squid qiant axons. The drug was applied internally, and decreases in peak transient and steady-state conductances as measured by voltage clamp technique were taken as an index of activity. When the total internal ketamine concentration was maintained constant, the suppression peak transient and steady-state conductances decreased with an increase in internal pH from 7.0 TO 8.4. When the concentration of the internally present charged form of ketamine was kept constant, the suppression of the peak transient conductance remained almost constant at internal pH values of 7.0, 7.3 and 7.7, but increased at pH 8.4. However, the suppression of the stead-state conductance became more prominent as the pH was raised from 7.0 to 8.4. With a constant internal concentration of the uncharged form, the suppression of both conductances decreased as the pH was raised from 7.0 to 8.4. Computation of dissociation constants to suppretamine is more potent than the charged form at all internal pH values examined. These data also show that the potency to suppress the peak transient conductance by the charged and uncharged forms of ketamine decreased as the intertance of the charged form increased, and that of the uncharged form decreased considerably with increase in the internal pH.


Uptake of uric acid by separated renal tubules of the rabbit. I. Characteristics of transport. A rapid filtration procedure was used to determine rates of uric acid uptake by a preparation of separated renal cortical tubules of the rabbit. The rate of uric acid uptake was temperature dependent and showed saturation kinetics with a K of 3.2 mM. The uptake was 50% lower when measured under nitrogen as compared with uptake under oxygen. The uptake rate increased with increasing sodium concentration and decreased with increasing potassium concentration. Uptake was stimulated by citrate, succinate and pyruvate, and inhibited by alpha-ketoglutarate. Parathyroid hormone and 10(-4) M adenosine 3':5'-monophosphate increased the uric acid uptake rate when preincubated with the tubules for 130 minutes before the addition of uric acid. Uric acid uptake in this preparation appears to occur by some form of carrier-mediated active transport.


Further evaluation of the discriminative effects of morphine in the rat. Rats were trained in a two-choice discrete trial avoidance paradigm to discriminate between saline and 3.0 mg/kg of morphine. Behavior was considered to be under stimulus ocntrol when the rats completed at least 90% of the trials in a 20-trial session on the morphine-appropriate choice lever after receiving morphine and when they completed at least 90% of the trials on the saline-appropriate choice lever after receiving saline. The discriminative effects of morphine, measured by responding on the morphine-appropriate lever, were then evaluated by determining the dose-response characteristics of representative narcotic analgesics, analgesics with mixed agonist and narcotic antagonist properties and nonopioid psychoactive drugs. Eight narcotic analgesics each produced dose-related responding on the morphine-appropriate lever. The relative potency for producing discriminative effects equivalent to those produced by 3.0 mg/kg of morphine ranged form etonitazene = 1000 x morphine to propoxyphene = 0.0175 x morphine. Of the narcotic antagonist analgesics tested, butorphanol and nalmexone produced discriminative effects equivalent to those of the morphine training dose whereas nalorphine, levallorphan, oxilorphan, nalbuphine and ketocyclazocine did not. The nonopioid psychoactive drugs, mescaline, ketamine, physostigmine and scopolamine, also failed to produce discriminative effect equivalent to those produced by 3.0 mg/kg of morphine. These results confirm and extend our previous findings that of those drugs which have also been evaluated in man, discriminative effects equivalent to the training dose of morphine are produced uniquely by narcotic analgesics and narcotic antagonists which produce morphine-like subjective effects. These results are compatible with the hypothesis that the properties of morphine which enable it to function as a discriminative stimulus in the rat are analogous to those responsible for producing subjective effects in man.


Discriminative effects of morphine in the squirrel monkey. Squirrel monkeys were trained in a two-choice discrete trial avoidance task to discriminate between intramuscular injections of saline and 3.0 mg/kg of morphine. Morphine (0.1-10 mg/kg) produced a dose-related increase in the number of trials completed on the morphine-appropriate lever. The stimulus control produced by the discriminative effects of morphine met the following criteria for classification as a specific narcotic effect: 1) morphine-like stimulus control was produced by all other narcotic analgesics tested (fentanyl, oxymorphone, levorphanol, methadone and meperidine); 2) in so doing, these drugs spanned a 900-fold potency range relative to morphine; 3) stimulus control was blocked by the specific narcotic antagonist naloxone; and 4) stereospecificity was a requirement for stimulus control--levorphanol produced stimulus control equivalent to 3.0 mg/kg of mrophine but its optical isomer dextrorphan did not. The time course of the stimulus control produced by 3.0 mg/kg of morphine showed that the animals continued to respond on the morphine-appropriate lever up to 14 hours after morphine administration. In contrast, monkeys administered 0.01 mg/kg of fentanyl responded on the morphine lever for only as lone as 1/2 hour after fentanyl administration. Naloxone, d-amphetamine and pentobarbital all failed to substitute for morphine, Thus, this study has extended previous observations of the discriminative properties of morphine in rats by demonstrating that qualitatively similar data are produced in a second species, the squirrel monkey.


Control of hepatic and intestinal blood flow: effect of isovolaemic haemodilution on blood flow and oxygen uptake in the intact liver and intestines. 1. Limited isovolaemic haemodilution was produced in cats by addition of dextran 75-Ringer solution to an extracorporeal blood reservoir connected in series with the cat. Total hepatic venous outflow was neasured using a hepatic venous long-circuit and hepatic arterial flow was measured with an electromagnetic flow probe. Oxygen uptake was monitored in the guts and liver. Na-pentobarbitone anaesthesia was used. 2. Following reduction of the haematocrit (from 31 to 22) the oxygen uptake of the gut segment and liver were maintained. Gut conductance increased to 125% of control while the oxygen extraction ratio increased to only 109%. The hepatic arterial conductance did not change in spite of a greatly reduced (to 68%) oxygen delivery. Hepatic extraction increased to 140% of control. 3. The hepatic artery did not dilate to maintain constant oxygen supply to the liver thus confirming our previous observation that blood flow is not coupled to hepatic metabolism. 4. Oxygen extraction in the gut correlated well with changes in portal blood flow but not with changes in vascular conductance, arterial blood pressure or oxygen delivery. 5. The blood flow of the gut (vascular beds draining into the portal vein in the splenectomized preparation) was controlled in a manner that prevented changes in portal venous PO2 in spite of a reduction in oxygen content. Local PO2 and perhaps pH, are suggested as the factors controlling gut blood flow following haemodilution. 6. Changes in portal blood flow correlated with changes in portal vascular (intrahepatic) conductance such that increased portal flow produced an increased portal conductance thereby maintaining portal venous pressure constant.


The measurement of distortion: theoretical considerations. When very find measurements are recorded for the purpose of establishing distortion values, from the standpoint of accuracy, specific procedures must be developed. In the procedural system defined here, a potential major source of error is indicated and the means of overcoming this error established. From the standpoint of modern electronic technology, the use of an analogue-to-digital interface with a digital computer is considered extremely advantageous and allows the utilization of technical help to carry out the preliminary measurements. Under these circumstances, major error sources, such as reader error and calculation error, are eliminated. Further, multiple readings on each point can be made without significant sacrifice of time. This latter statement can help define the reading accuracy on each point and allows an immediate further set of readings on any point that has a large variation in the initially established set of readings. The applicability of this approach to any system in which an accurate set of readings is required to define distortion or some other parameter is limited only by the imagination of the investigator. Measuring points can be established at any desired location, and the number required is a function of the mathematical analysis being attempted. The use of a digital computer to compute the final values allows an immediate remeasurement of the initial values if necessary, since the computer response time is of the order of seconds for the calculations carried out. This is very important if changes in the experimental setup are likely to occur with time. In general, a computerized scheme such as is proposed here allows the investigator to concentrate on the experiment rather than the calculation of results. Also, the use of technical help to carry out delicate measuring procedures can allow the investigator more time to analyze the final results.


Cathepsin D activity in bovine articular cartilage, synovial membrane and fluid: degradation of cartilage proteoglycans from same joint. Cathepsin D type proteases were extracted from articular cartilage, synovial membrane, and synovial fluid from normal, adult bovine knee joints. A sensitive enzyme assay made it possible to measure protease activity in the different tissues from individual joints. Highest activity was found in the synovial membrane, while cell free synovial fluids contained comparatively low activity. The degrading effect on articular cartilage proteoglycans (PGC and PGS), isolated from the same joints, was demonstrated by gelfiltration on Sepharose columns and by viscometry. Gelfiltration profiles of incubation mixtures indicated a proteolytic effect on PGC and on PGS), at pH 3.5, in concentrations of enzyme and proteoglycans found in cartilage tissue. No effect at neutral pH was obtained despite a 100-fold increase of enzyme concentration. These findings were supported by viscometry data. The degrading effect of enzymes from all sources was completely inhibited by pepstatin.


3-Halo-5,7-dimethylpyrazolo [1,5-a]pyrimidines, a nonbenzodiazepinoid class of antianxiety agents devoid of potentiation of central nervous system depressant effects of ethanol or barbiturates. Forty derivatives (1-40) of pyrazolo[1,5-a]pyrimidine were synthesized and evaluated for antianxiety properties via gross behavioral observations in rats. Five of these compounds, including 5,7-dimethylpyrazolo[1,5-a]pyrimidine (6) and the 3-fluoro (7), 3-chloro (8), 3-bromo (9), and 3-iodo (10) derivatives, were selected for advanced evaluation. Although 6 and 7 had marginal activity, 8-10 had an anxiolytic effect in animals comparable to the clinically useful benzodiazepines, diazepam, and chlorodiazepoxide. Comparison with chlorpromazine indicated that 6-10 are probably not antipsychotic agents. These compounds also lacked activity in anticonvulsant and analgesic tests. Acute toxicity data (mouse, ip and po) indicated that 8-10 had excellent therapeutic ratios, although 10 was more poorly absorbed than 8 and 9. Further demonstration of anxiolytic efficacy was obtained by comparing the effects of 8 and 9 with the benzodiazepines in modifying provoked aggression in monkeys, rats (muricide), and fighting mice. The most remarkable observation, however, was that 8 and 9 had no effect, at the anxiolytic threshold, in potentiating the CNS depressant effects of ethanol or sodium barbital (po) in treated mice. In contrast, diazepam and chlorodiazepoxide potentiated this drug interaction effect at minimal anxiolytic doses.


Synthesis and adrenergic beta-blocking activity of some 1,3-benzodioxole derivatives. A series of 1,3-benzodioxole derivatives was synthesized. We found four compounds (2,3,10 and 11 in Table IV) to have about the same order of beta-blocking activity as that of sotalol. In addition, it is of interest that some of the compounds (2-4) were found to have hypotensive activites, although they were about one-tenth of that of hydralazine. Sotalol did not produce any change in blood pressure, and propranolol raised the blood pressure.


Beta-adrenoceptor studies. 2. Effects of alkyl substitution on beta-adrenoceptor blocking, antiarrhythmic, and local anesthetic activities of 1,1'-(o-phenylenedioxy)bis(3-isopropylamino-2-propanol). A series of bis(2-hydroxy-3-isopropylaminopropyl) ethers of nuclear-substituted catechols (1-7) has been synthesized and examined in vitro for beta-adrenoceptor blocking activity, antagonism of ouabain-induced arrhythmias, and local anesthetic activity. Both tracheal and right atrial beta-adrenoceptor blocking activity are markedly decreased by alkyl substitution in position 3 of parent catechol diether 1. Substitution in position 4 still lowers the affinity to cardiac arrhythmias and local anesthetic activity increases with introduction of alkyl substituents in the 3 as well as in the 4 position. In contrast with biological activities, the partition coefficient 1-octanol-phosphate buffer, pH 7.40, of 1 did not change significantly by 3- and 4-methyl substitution. Stepwise multiple regression analyses were performed using log P or pi values in combination with pKa(m), E8, or sigma. With cardiac beta-adrenoceptor blocking activity the optimal equation contained E8 and pi parameters, tracheal activity appeared to depend mainly on the E8 parameter, whereas for antiarrhythmic and local anesthetic activities the lipophilicity of the substituents appeared to be the determinant factor.


Synthesis and hypoglycemic activity of S-acyl derivatives of 3-mercaptopicolinic acid. A series of S-alkanoyl and benzoyl derivatives of 3-mercaptopicolinic acid (3-MPA) was prepared and studied for hypoglycemic activity. Three alkanoyl derivatives (propionyl, pivaloyl, and 1-adamantanecarbonyl, 19-21) were prepared with increasing bulk around the thio ester bond. The benzoyl derivatives contained aromatic substituents chosen from a sigma-pi cluster chart so that the esters prepared had a wide range of electronic and solubility properties. In general, compounds with substituents which increased lipid solubility [p-chlorobenzoyl (4), p-trifluoromethylbenzoyl (6), and pivaloyl (20)] had the greatest potency at a dose of 300 mg/kg. Hydrolysis rates, measured at pH 6 and 8, indicated that in vivo breakdown to 3-MPA probably did not account for the observed hypoglycemic activity of the esters. 4, 6, and 20 were less potent than 3-MPA in comparative dose range studies.


Relative concentrations of zwitterionic and uncharged species in catecholamines and the effect of N-substituents. The relative concentrations of zwitterionic and uncharged species for the series of N-substituted catecholamines (I, R1 = R2 = OH; R = H, Me, Et, i-Pr, t-Bu) are derived from the pKa data published in 1962 by Sinistri and Villa. The concentration ratios, represented by the tautomeric equilibrium constant Kt, show a definite trend and are respectively 1.8, 4.3, 4.7, 4.7, and 7.1. These values suggest that any mechanism of action involving proton transfer, which might transform the zwitterion into the uncharged form, would be most favorable for norepinephrine and least favorable fo the t-Bu derivative.


Interviewing skills: a comprehensive approach to teaching and evaluation. Evaluation procedures utilizing standardized interviews scored by means of a formal objective rating system which make it possible to measure the interviewing skills of health professional students have been developed in recent years. These procedures have been used in the Child Health Associate Program at the University of Colorado Medical School to evaluate the ability of a group of students to achieve the objectives of a practice-oriented interviewing course. Results from the standardized interviews indicate that students taking the course gathered an average of 76 percent of all available data and used 86 percent of the process skills defined as necessary for positive interview interaction. A previous group of child health associate students who did not take the course gathered an average of 47 percent of all available data and used an average of 62 percent of the necessary process skills.


Evaluation of a child health associate program. The staff of the University of Colorado Child Health Associate Program critically reviewed the effectiveness of the program's structure and content during an intensive two-day seminar. The review was conducted through workshops involving participants representing students, graduates, faculty, employers, funding agencies, university administrators, and educational consultants. The agenda for the evaluation included workshops on specific topics such as basic and clinical sciences, psychosocial skills, and proficiency testing. Information obtained provided extremely valuable data which were used to improve the program.


A protonmotive force as the source of energy for galactoside transport in energy depleted Escherichia coli. An artificially produced electrochemical potential difference for protons (portonmotive force) provided the energy for the transport of galactosides in Escherichia coli cells which were depleted of their endogenous energy reserves. The driving force for the entry of protons was provided by either a transmembrane pH gradient or a membrane potential. The pH gradient across the membrane was created by acidifying the external medium. The membrane potential (inside negative) was established by the outward diffusion of potassium (in the presence of valinomycin) or by the inward diffusion of the permeant thiocyanate ion. The magnitude of the electrochemical potential difference for protons agreed well with magnitude of the chemical potential difference of the lactose analog, thiomethylgalactoside. The observations are consistent with the view that the carrier-mediated entry of each galactoside molecule is accompanied by the entry of one proton.


Studies on the cation permeability of human red cell ghosts. Characterization and biological significance of two membrane sites with high affinities for Ca. Net K movements in reconstituted human red cell ghosts and the resealing of ghosts to cations after osmotic hemolysis of red cells have been studied as functions of the free Ca ion concentration. The Ca-dependent specific increase in K permeability was shown to be mediated by a site close to the internal surface of the membrane with an apparent dissociation constant ap pH 7.2 for Ca (K'p1) of 3-5 X 10(-7) M, for Sr of 7 X 10(-6) M. Ba and Mg did not increase the K-permeability of the membrane but inhibited the Ca-mediated permeability changes. K'D1 decreased in a nonlinear fashion when the pH was increased from 6.0 to 8.5. Two different pK' values of this membrane site were found at pH 8.3 and 6.3. The Ca-activated net K efflux into a K-free medium was almost completely inhibited by an increase in intracellular Na from 4 to 70mM. Extracellular K antagonized this Na effect. Changes in the extracellular Na (0.1-140 mM) or K (0.1-6 mM) concentrations had little effect and did not change K'p1. The Ca-stimulated recovery of a low cation permeability in ghost cells appeared to be mediated by a second membrane site which was accessible to divalent cations only during the process of hemolysis in media of low ionic strength. The apparent dissociation constant for Ca at this site (K'p2) varied between 6 X 10(-7) and 4 X 10(-6) M at pH 7.2 Mg, Sr, and Ba could replace Ca functionally. The selectivity sequence was Ca greater than Sr greater than Ba greater than Mg. K'p2 was independent on the pH value in the range between 6.0 and 8.0 Hill coefficients of 2 were observed for the interaction of Ca with both  membrane sites suggesting that more than one Ca ion is bound per site. The Hill cofficients were affected neither by the ion composition nor by the Ph values of the intra-and extracellular media. It is concluded that two different pathways for the permeation of cations across the membrane are controlled by membrane sites with high affinities for Ca: One specific for K, one unspecific with respect to cations. The K-specific "channel" has properties similar to the K channel in excitable tissues.


Enhancement of 5-iododeoxyuridine-induced endogenous C-type virus activation by polycyclic hydrocarbons: apparent lack of parallelism between enhancement and carcinogenicity. When mouse MLg cells were treated with 3-methylcholanthrene or 7,12-dimethylbenz[alpha]anthracene in the presence of microsomal enzymes and NADPH after 5-iododeoxyuridine (IUDR) treatment, the induction rate of the endogenous C-type virus was increased fivefold to sixfold in comparison with the culture treated with IUDR only. In this reaction, both the microsomal enzymes and NADPH were indispensable. 7,8-Benzoflavone, an inhibitor of the metabolism of hydrocarbons in hamster embryo cultures, inhibited the reaction. For detecting the enhancing activity, the concentration of IUDR for the pretreatment, the concentration of the test products, and the duration of the treatment with the products were important factors. In screening 30 polycyclic hydrocarbons, we were unable to detect a correlation between the in vivo carcinogenicity in the skin and the enhancing activity in the conditions tested.


Lack of influence of hypophysectomy on estrogen-induced DNA synthesis in Leydig cells of BALB/c mice. In mice of strains susceptible to Leydig cell tumor induction, treatment with estrogens induced a "spurt" of DNA synthesis within the first few days. This synthetic activity generally subsided, until areas of Leydig cell hyperplasia developed several months later. Autoradiographic and quantitative biochemical studies indicated that in BALB/c mice this initial DNA synthetic activity occurred in the absence of the hypophysis and apparently was the result of effects of estrogen directly on Leydig cells. Although hypophysectomy inhibited sperm maturation, [3H]thymidine incorporation into spermatogonia was reduced only slightly 2 weeks after surgery, as was the induced DNA spurt in the interstitial tissues.


Effect of hyperthermia and environmental acidity on the proteolytic activity in murine ascites tumor cells. The influence of hyperthermia and environmental pH on proteolytic activity was studied in murine ascites tumor cells in vitro. PNJ ascites tumor cells were incubated with [125I]cytochrome c at 42.5 or 37 degrees C in a modified Krebs-Ringer buffer adjusted to pH 7.2 or 6.4. Incubation at normal temperature at pH 6.4 and 7.2 or at 42.5 degrees C and pH 7.2 resulted in identical protein digestion. However, hyperthermic incubation at pH 6.4 resulted in a significant increased activity. This was also observed after only 1 hour of hyperthermic incubation followed by subsequent incubation in an acidic environment at normal temperature. The increased proteolytic activity following hyperthermic treatment under acidic conditions may support the hypothesis that increased lysosomal activity is of primary importance in the hyperthermic tumor-cell destruction in vivo.


Nature of the delayed graft-versus-host reactivity of fetal liver cell transplants in mice. Experiments were designed to determine which actual differences in the cellular composition between fetal liver and bone marrow account for the distinct types of graft-versus-host (GvH) disease. The assay of reactive lymphocytes (by in vitro mitogenic stimulation) in fetal liver transplants in mice, the purification of hemopoietic stem cells (HSC) of the transplants, and the quantitation of HSC numbers in the grafts traced the basis for the distinctly weak type of GvH disease after fetal liver cell grafts. It was found that transplantation of purified HSC concentrates did not modify the severity of GvH mortality. The moderate character of the delayed GvH disease was shown to depend on the presence of an HSC population in fetal liver with different qualities and not on numerical differences between the HSC in fetal liver and bone marrow. Data collected also demonstrated that when GvH disease occurred in the recipients of transplants of fetal liver, it shared the characteristic histologic features of the bone marrow GvH syndrome. The recovery of mitogen responsiveness of spleen cells may have been delayed in fetal liver allotransplantation as compared to syngeneic grafting. By supportive infusion of lymphoid cells, it was suggested that the immunodeficiency coinciding with GvH disease represented a secondary manifestation of the disease rather than a primary impairment in lymphoid differentiation.


Characterization of group H streptococcal temperate bacteriophage phi 227. phi 227, a temperate phage from a group H streptococcus (Streptococcus sanguis), was propagated vegetatively in group H strain Wicky 4-EryR, and its characteristics were determined. A procedure dependent on multiplicity of infection, incubation time, and treatment of crude lysates with diatomaceous earth was found to optimize phage yield, resulting in titers of 1 X 10(10) to 2 X 10(10) PFU/ml. Without prior treatment with diatomaceous earth, subsequent purification procedures (methanol, ammonium sulfate, polyethylene glycol) gave recoveries of less than 1% of crude lysate titers. Adsorption of phi227 to host cells was relatively unaffected by the medium, but calcium (not substituted by magnesium) was required for formation of infectious centers. The phage receptor was present on purified cell walls, resisted trypsin and heat, and was removed ty hydrochloric acid, trichloracetic acid, and hot formamide: however, formamide-extracted material failed to inactivate phage, and the nature of the receptor is unknown. Single-step growth experiments showed a latent period of 39 min and a burst size of 100 PFU/infectious center; results were unaffected by omission of supplemental Ca2+, by supplementation with Mg2, addition of glucose, or changes of pH between 6.35 and 8.0; but increased temperature (40 to 43 degrees C) shortened the latent period and decreased the burst size. The latent period was prolonged in genetically competent host cells and in chemically defined medium; and in the latter, the burst size was smaller. Phage replication was sensitive to those metabolic inhibitors which inhibited the host streptococcus: these included rifampin, fluorodeoxyuridine, hydroxyurea, dihydrostreptomycin, and 6-P-hydroxyphenylazouracil. The data suggest that phi227 does not code for a rifampin-resistant RNA polymerase. However, in a rifampin-resistant host strain, phage replication and lysogen formation were both decreased suggesting that altered host core polymerase had less affinity for (some) promotors on the phi227 template. In transfection, a Ca2+-dependent stabilization step that was inhibited by Mg2+ was demonstrated; transformation was not affected by either Ca2+ or Mg2+, and the site and nature of the stabilization are unknown. More than one molecule of DNA was required for plaque formation. Biophysical characterization showed a type B phage of buoyant density (CsCl) 1.50, containing five proteins and 54.8% DNA. The duplex linear DNA had a molecular weight (calculated from contour length) of 23.2 X 10(6) and a guanine plus cytosine content (calculated from melting point) of 42.3 mol%. Similar characterizations of streptococcal phages, including biophysical data, have not been previously available.


Characterization of components released by alkali disruption of simian virus 40. Treatment of simian virus 40 (SV40) particles at pH 9.8 in the presence of 1 mM dithiothreitol for 5 min at 37 degrees C disrupted the virions into a 60S DNA-protein complex and DNA-free 7S protein particles. The DNA-protein complex contained approximately equal amounts of DNA and protein, and appeared by electron microscopy to be relaxed circular structures with an average of 21 beads joined by short, thin bridges. The major protein components in the complex were host cell histones, but SV40-specific proteins VP3 and VP2 were also present. The 7S protein particles were almost exclusively VP1 and, in negatively stained samples, resembled the capsomer structures of intact virions.


Temperature-sensitive defect of vesicular stomatitis virus in complementation group II. The prototype member of the complementation group II temperature-sensitive (ts) mutants of vesicular stomatitis virus, ts II 052, has been investigated. In ts II 052-infected HeLa cells at the restrictive temperature (39.5 degrees C), reduced viral RNA synthesis was observed by comparison with infections conducted at the permissive temperature (30 degrees C). It was found that for an infection conducted at 39.5 degrees C, no 38S RNA or intracytoplasmic nucleocapsids were present. For nucleocapsids isolated from ts II 052 purified virions or from ts II 052-infected cells at 30 degrees C, the RNA was sensitive to pancreatic RNase after an exposure at 39.5 degrees C in contrast to the resistance observed for wild-type virus. The nucleocapsid stability of wild-type virus when heated to 63 degrees C or submitted to varying pH was not found in nucleocapsids extracted from ts II 052 purified virions. The data suggest that for ts II 052 there is an altered relationship between the viral 38S RNA and the nucleocapsid protein(s) by comparison with wild-type virus. Such results argue for the complementation group II gene product being N protein, so that the ts defect in ts II 052 represents an altered N protein.


Receptor interaction between eastern equine encephalitis virus and chicken embryo fibroblasts. The attachment of eastern equine encephalitis virus to chicken embryo fibroblasts was studied at 0 degrees C. The binding specifically responsible for initiating infection was studied in the initial experiments by employing plaque-forming ability as the measured response. Results from these initial studies were closely paralleled in studies of binding of radiolabeled virus under the same conditions. Binding that had occurred at the pH optimum, pH 6.5, could be reversed only at higher pH. The observed pH dependence of virus attachment suggested the interaction of at least two ionizable species in the initial binding of virus to cell, and that one to three attachments must occur between virus and cell prior to infection.


Priapism: evolution of management in 48 patients in a 22-year series. The choice of an effective method to treat priapism is challenging because precise causes in the majority of patients have not been well defined. A review of 48 patients treated during a 22-year period shows evolution of a regimen of management that has yielded a high percentage of success. Idiopathic priapism and sickle cell disease accounted for 81 per cent of the subjects. An evaluation should include a medication history, a search for specific diseases, as well as a thorough physical examination to detect possible etiologic factors. The explanation for the frequent association of fever deserves further investigation. Initial therapy consisting of aspiration and irrigation, and intermittent pneumatic cuff compression should be undertaken for a trial period of 12 to 36 hours, repeating the aspiration 2 or 3 times if necessary. The failure of priapism to resolve after such treatment is an indication for a shunt operation. Patients with known etiology should be treated specifically for the primary disease and usually more conservatively for priapism. Resolution occurred in all patients and approximately 50 per cent regained sexual potency.


Venographic localization of the non-palpable undescended testis in children. In cases of bilateral non-palpable undescended testes in which human chorionic gonadotropin stimulation has shown the presence of testicular tissue and in cases of unilateral non-palpable undescended testes selective transfemoral gonadal venography with a modified Seldinger technique has been used for the preoperative localization of the non-palpable testis. since the undescended testis may be located anywhere along the course of its embryologic descent, that is from the level of the renal fossa to its exit from the inguinal canal, preoperative localization will aid in the surgical management. Gonadal venography has proved to be accurate and safe, and has aided in the determination of the extent of surgical exploration in 9 children with 12 non-palpable undescended testes (6 right and 6 left).


The effects of glucose infusion on myocardial performance during acute hypoxia. The effects of hypoxia with or without glucose infusion on the cardiac contractility, blood pressure, electrocardiogram, blood electrolytes (sodium and potassium), glucose, pH, Po2, and Pco2 in anesthetized dogs were studied. Hypoxia was induced by ventilating the dogs with reduced oxygen (10%) in the inspired air. Hypoxia produced a decrease in the cardiac contractility and blood pressure, and an increase in the heart rate and central venous pressure. It produced a decrease in the blood pH, Po2, and Pco2, and an increase in the blood glucose and potassium. Glucose infusion during hypoxia delayed the rate of decrease in the contractility and blood pressure significantly. The time for decrease in the contracility to 45 to 50% was increased by 67%. Glucose infusion prevented the loss of potassium from the cell. Glucose infusion however, was unable to correct acidosis. These results indicate that glucose infusion during hypoxia might prevent or delay the deterioration of myocardial function.


Effects of beta-adrenergic blocking agents on the blood pressure, plasma renin activity and hemodynamics of hypertensive patients. Changes in blood pressure, plasma renin activity, and hemodynamic components were studied in 23 patients with essential hypertension treated with oral pindolol or propranolol. These beta-adrenergic blocking agents effectively lowered the blood pressure in the majority of the patients. Although plasma renin activity was not significantly changed, the higher was the pretreatment level, the more it tended to be decreased. Systemic vascular resistence was significantly decreased, while changes in cardiac index and circulating blood volume were variable. Pindolol showed less effect in reducing the heart rate than propranolol. The antihypertensive effect of these drugs had no correlation with the change in plasma renin activity or in any one of hemodynamic components.


[Metabolism of tetrachloroethylene in guinea pigs (author's transl)]. Tetrachloroethylene oxide was chemically prepared from tetrachloroethylene, and the metabolites of the oxide in guinea pigs were analyzed by gaschromatography and Fujiwara reaction. The results obtained are as follows: 1) Trichloroacetic acid appeared in gaschromatogram after injection of tetrachloroethylene oxide, but trichloroethanol did not. 2) The metabolites analyzed by Fujiwara reaction after injection of tetrachloroethylene oxide were composed of large proportion of trichloroacetic acid and small proportion of trichloroethanol. 3) The ratio of trichloroacetic acid to trichloroethanol in the urine in case of tetrachloroethylene oxide was relatively similar to that of tetrachloroethylene. 4) The effects of pH (2.0 and 10.0) and temperature (4 degrees C and 37 degrees C) on the urinary metabolites suggest that the substance equivalent to trichloroethanol by Fujiwara reaction in metabolites may not be a real one. 5) Toxicity of tetrachloroethylene oxide seems to be much higher as compared with that of tetrachloroethylene in consideration of the maximum allowable dose in guinea pigs. 6) It is supposed from our experiments that tetrachloroethylene oxide is an intermediary metabolite of tetrachloroethylene.


Serum gamma-glutamyl transpeptidase as a diagnostic aid in the periodic health examination. Serum gamma-glutamyl transpeptidase is one of the enzymes in diagnosis of liver diseases, since a new colorimetric method was devised by Orlowski, M. et al. Forty mU/ml of serum gamma-glutamyl transpeptidase activity is said to be the upper limit at clinical level. When this value is set up as a screening level in the periodic health examination, about 35% of the subjects including daily drinkers can be evaluated as abnormal. In the present study, the upper limits of serum gamma-glutamyl transpeptidase activity were 102 mU/ml for 147 normal subjects including daily drinkers and 49 mU/ml in 70 non-drinkers selected from the subjects. Therefore, we propose that the standards for screening the abnormal from the normal in the periodic health examination should be 50 mU/ml for non-drinkers and 100 mU/ml for drinkers.


[Studies on the adsorption removal of ammonia gas. 1) Adsorption of ammonia gas on several kinds of activated carbons (author's transl)]. The present study was designed to secure some fundamental informations on the adsorption removal of ammonia gas by the static method. In order to find out the suitable activated carbons for the adsorption removal of ammonia gas, amounts of ammonia gas adsorbed on twelve kinds of activated carbons were measured at 30 degrees C and up to 70,000 ppm of ammonia gas. The relations between the amounts of ammonia gas adsorbed on the activated carbons and the physical properties of them were discussed through the results of specific surface area, pore volume, mean pore radius, scanning electron micrograph, pH, and amount of base. The results were as follows: 1) Among the twelve kinds of activated carbons, the activated carbons No. 2, No. 3, and No. 6 adsorbed larger amount of ammonia gas than the others. 2) Adsorption of ammonia gas on the activated carbons seemed to be mainly physical as judged from the values of heat of adsorption. 3) The adsorption capacity of the used activated carbons can be recovered to the original capacity by some treatment. 4) The period to reach adsorption equilibrium was about 5 minutes. 5) It may be concluded that adsorption of ammonia gas on the activated carbons was decided mainly by the surface properties  (pH and amount of base) of the activated carbons rather than their porous structures.


Effects of bufetolol and propranolol on active and passive membrane properties of dog papillary muscle. Effects of bufetolol and propranolol, adrenergic beta-receptor blocking and anti-arrhythmic drugs, on active and passive membrane properties of the dog papillary muscle were investigated with microelectrode and sucrose-gap methods. Bufetolol (10(-5) to 10(-4) g/ml) and propranolol (10(-6) g/ml) significantly decreased the maximum rate of rise of the action potential. The maximum responsive frequency to driving stimulus was decreased in the presence of bufetolol (3 X 10(-5) g/ml) and propranolol (10(-5 g/ml), whereas the effective refractory period was not affected. The critical threshold potential was shifted to more positive potential in the presence of the drugs. The passive membrane property, the space constant (lambda), the time constant (tau) and the current-voltage relations of the muscle membrane were not significantly altered by the drugs. It is concluded that bufetolol and propranolol suppress the excitability of the muscle membrane and this action may be ascribed to the decrease in the sodium conductance (gNa) and to the rise of gNa onset potential without alteration in the passive membrane property.


Effects of L-glutamine on acetylsalycylic acid induced gastric lesions and acid back diffusion in dogs. Effects of L-glutamine on acetylsalicylic acid (ASA)-induced gastric mucosal lesions were studied in mongrel dogs. It was confirmed that when oral ASA at 1.0 or 2.0 g per dog is given in two divided doses, there is severe and consistent dose-dependent mucosal damage in the glandular portion of the stomach in fasted dogs. However, when L-glutamine 2.0 or 4.0 g per dog in two divided doses is given concomitantly with ASA 2.0 g per dog orally, the gastric irritation is significantly inhibited. Instillation of 20 mM of ASA in 100 mM HCl solution into the Heidenhain pouch of Beagle dogs produced a significant loss of H+ from the pouch and a gain of Na+ in the lumen compared with ASA-free controls. When L-glutamine (100 mM) was given concomitantly with ASA (20 mM) into the pouch, changes of electrolyte fluxes in response to ASA alone were significantly suppressed. However, 50 mM of L-glutamine had no appreciable effect on acid back diffusion caused by ASA 20 mM. The amino acid itself had little effect on the ionic movement in the pouch. Gross bleeding from the pouch treated with ASA was never observed with the concomitant dosing of ASA and L-glutamine 50 or 100 mM.


Complement-independent nephrotoxic nephritis in the guinea pig. Immunologic mechanisms of proteinuria were investigated in guinea pigs (GP) injected with sheep antiserum (NTS) to GP glomerular basement membrane (GBM). Linear deposition of sheep gamma 1 and gamma 2 IgG led to a prompt but transient (36 hr) increase in albumin excretion from control values of 0.026 +/- 0.013 mg/hr to maximal values of 26+/-12.1 mg/rh at six hours without detectable histologic or electron microscopic changes except for decreased staining for glomerular polyanion and epithelial cell foot process fusion. GBM permeability to anionic ferritin was not increased during proteinuria. Anti-GBM antibody deposits did not fix GP C3 or C4 in vivo or in vitro. NTS-induced proteinuria was the same in guinea pigs that were normal, greater than 95% depleted of C3 through C9, genetically deficient in C4, and depleted of circulating polymorphonuclear leukocytes (PMN). Prior administration of antihistamines, steroids, azathioprine, colchicine, indomethacin, heparin, aprotinin (Trasylol), and niridazole also failed to reduced proteinuria. Initial proteinuria subsided by 36 hr, did not recur despite linear deposition of GP gemma 1 and gemma 2 after day seven, and antibody to GMB-bound sheep globlin. In the GP nephrotoxic nephritis model, anti-GBM antibody deposits apparently mediate increased permeability to albumin by a currently undefined mechanism which is independent of complement, PMN, and other know mediators of inflammation.


[Chloroquine keratopathy as an example of drug-induced phospholipidosis (contribution to the pathogenesis of cornea verticillata) (author's transl)]. Chronic treatment with certain drugs induces morphological alterations in the eye which are histologically and electron microscopically identical with those found in hereditary lipidoses (cornea verticillata, e.g. in M. Fabry). Hereditary storage diseases are the consequence of enzyme defects, the mechanism underlying the side effect of certain drugs, however, is quite different. "Amphiphilic" drugs from completely different pharmacological groups, like chloroquine, amiodarone, chlorpromazine form complexes with cellular phospholipids which cannot be metabolised by lysosomal phospholipases. Thus in all tissues with high phospholipid content or turnover typical intracellular deposites with lamellary or crystalloid structure may occur (myelin figures). Such deposites were observed in different parts of the eye and are known e.g. from the cornea as "cornea verticillata".


Carbon tetrachloride-induced changes in mixed function oxidases and microsomal cytochromes in the rat lung. The effects of a single exposure, by gastric intubation or inhalation, to carbon tetrachloride (CCL4) on rat lungs were assessed. By 1 to 7 days, focal areas of alveolar collapse, septal edema, and modification of type II pneumonocytes were observed. By 24 hours after exposure to the toxin, there were no identifiable changes in surfactant levels or distribution. Microsomes obtained from the lungs and prepared for analysis revealed marked decreases in cytochrome P-450 content and P-450-related N-demethylation of dimethylaniline. Only a transient reduction of cytochrome b5 occurred, with a rebound exceeding control values during the period of pulmonary healing. Whether the lung acted as an excretory route (following intubation) or as an absorption path (after inhalation) made little difference. Carbon tetrachloride had no effect on in vitro microsome composition and function unless supplemented with a reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) generating system. Under these circumstances, there was a reduction in both cytochromes b5 and P-450. Our data indicate that a considerable chemical modification of the pulmonary tissues had taken place, with no accompanying easily recognized changes in cellular structure. Furthermore, evidence for the in vitro destruction of pulmonary microsomal cytochromes P-450 and b5, unrelated to peroxidation, is indicated by these findings.


Acid hydrolases. Assay of activity and latency in the varied mixed cell populations of canine gastric mucosa. The specific enzymic properties, membrane or particle binding capacities, and the total activities of certain acid hydrolases, including cathepsin D, acid phosphatase, arylsulfatase, and five acid glycosidases have been compared in normal canine antral and fundic mucosae and in liver. The two major regions of the gastric mucosa, whose cell populations are comparable in type but have very distinct functions, also differ in many properties of their lysosomal enzymes. These differences necessitate several major modification in their method of assay. Using optimal conditions, the activities of most of these enzymes were found to differ: levels in the antrum, in spite of its high water and mucin-glycoprotein content, were significantly greater, suggesting that the high lysosomal hydrolytic activity may be associated with the rapid autophagic processes of normal turnover of its surface epithelial and mucous neck cells. Lysosomal membrane stability or latency is also greater in the antrum; this may account, in part at least, for antral resistance to erosions brought about by stress.


An evaluation of the immune state in leprosy. An evaluation of the immune state in leprosy was done by the application of a system of graft-versus-host reaction. Peripheral blood lymphocytes obtained from patients with different forms of leprosy and from normal healthy individuals were injected intravenously into the irradiated mice. The rate of blast transformation of the donor cells was measured by the radio-active thymidine uptake. The number of cells labelled with tritiated-thymidine was much higher in the normal individuals and patients with tuberculoid leprosy than in the patients with lepromatous leprosy with the borderline group placed in between the two. However, following successful treatment with DDS, an increased responsiveness and active DNA synthesis could be observed in the previously less responsive lepromatous lymphocytes.


[Intestinal autoflora of the test subjects in a 6-month biological engineering experiment]. During a 6-month bioengineering experiment the intestinal microflora of four test subjects was examined. Changes in the composition of different groups of intestinal microflora (bifidobacteria, lactic-acid bacteria, sporogenous anaerobes, proteus, etc) were found. In spite of the unstable pattern of the intestinal microflora and its tendency for simplification, the total number of microorganisms in 1 g of feces remained relatively unchanged in all the test subjects.


[Sensitivity of animals to central nervous system stimulants in hypokinesia]. The experiments were carried out on 1150 non-inbred white male rats weighing 200+/-50 g. The animals were housed in small cages for 1, 5, 10, 15, 30, 45 and 60 days. Control rats remained normally active. The experimental and control animals were given a typical diet. On the above days the rats were injected intraperitoneally with central nervous stimulants--caffeine, phenamine and strychnine. Changes in the animal sensitivity to the stimulants were measured with respect to the alterating of LD16, LD50 and LD54 in test animals as compared with the controls and in regards to the emergence and duration of behavioural reactions: adynamics (caffeine), stereotype behavior (phenamine) and convulsions (strychnine). The greatest changes in the animal sensitivity were noted in response to phenamine. A significant increase in the sensitivity to caffeine was found on the 5, 15 and 45th experimental days and to strychnine only on the 5 and 45th days. Convulsions in response to strychnine were recorded in experimental animals earlier than in the controls and their duration was dependent on the doses injected. Adynamics in response to caffeine developed in experimental rats later than in the controls (on the 15th day) and its duration changed cyclically. Stereotype behavior in response to phenamine showed cyclic pattern and its duration in experimental rats was shorter than in the controls.


Immunohistochemical studies on the localization and distribution of monoamine neuron systems in the rat brain II. Tyrosine hydroxylase in the telencephalon. Extensive plexuses of TH-positive nerve terminals were found in many parts of the telencephalon, mainly confined to the subcortical and limbic cortical structures. Of special interest were the distinct networks of varying densities in the amygdaloid cortex, the entorhinal cortex, the prepiriform cortex, the anterior cingulate cortex and the (pre-)frontal cortex. Their distribution is identical with the patterns observed in recent studies on cortical dopamine nerve terminals using certain modifications of the Falck-Hillarp technique. The extremely dense TH innervations patterns of the caudate nucleus, nucleus accumbens, tuberculum olfactorium and the less dense basket-like innervation of the lateral septal nuclei could also be demonstrated. TH-positive cell bodies in a periglomerular position could be observed in the olfactory bulb. A few TH-positive cell bodies were observed in the area around the anterior commissure and in the cingulate cortex. In one area, the hippocampal formation, TH-positive dotlike structures were located in the position of the mossy fibres. In all probability they do not belong to monoamine neurons but may contain a cross-reacting protein. In general, the distribution and density of TH-positive terminals agrees well with extensive regional, biochemical studies on TH activity performed by other groups. Minor discrepancies are discussed. As stated in a parallel study on the distribution of TH in the mes- and diencephalon these findings indicate that TH activity is closely related to the amount of enzyme protein. The TH enzyme levels seem to be much higher in the DA than in the NA nerve terminals of the forebrain which would explain the preferential demonstration of DA terminals in the forebrain using TH antiserum and the high and low TH enzyme activity in DA and NA rich regions, respectively.


pH-dependency in diffusion of some weakly acidic drugs, warfarin, sulfaethidole, and barbital, into organic phases. The diffusion of three weakly acidic drugs (warfarin, sulfaethidole, and barbital) in cyclohexane-buffer and n-octanol-buffer systems was studied by two different methods; shaking and model cell type of experimentation. At low pH values the drugs moved generally more readily into the organic phases than at higher pH. Moreover, when the shaking method was used the drugs moved readily to the n-octanol phase even at higher pH levels. Lecithin improved the diffusion at low more than at high pH levels. Thus, the results varied according to the organic phase selected and the method used.  Poor correlation was found between some results of this study and previously reported results of in situ absorption from the rat gastrointestinal tract of the same drugs. The results do not support the assumption that phospholipids may have a role in the absorption of ionized moieties.


Involvement of cytochrome b5 in the oxidative desaturation of linoleic acid to gamma-linolenic acid in rat liver microsomes. The effects of antibodies against microsomal electron-transport components on the in vitro activity of delta6-desaturation of linoleic acid to gamma-linolenic acid have been studied in intact microsomal membranes of rat liver. Reduced nicotinamide adenine dinucleotide (NADH) or reduced nicotinamide adenine dinucleotide phosphate (NADPH) (0.87 mM) served as electron donors, and effectively prompted the delta6-desaturase activities with yields of about 1.1 to 1.3 nmol per mg of protein in 10 min. Of the two antibodies studied under the same in vitro conditions, i.e., rabbit antisera preparations against rat liver microsomal hydrophilic parts of cytochrome b5 and NADPH-cytochrome c reductase, only the antibody against cytochrome b5 demonstrated a marked ability to inhibit the delta6-desaturase activity. This evidence supports a participation of cytochrome b5 in the delta6-desaturation of linoleic acid and suggests a pathway analogous to the delta9-desaturation of stearyl-CoA.


Glycerokinase in human adipose tissue.

Glycerokinase in human adipose tissue. The presence of glycerokinase has been demonstrated in human omental and subcutaneous adipose tissue. The enzyme reaction showed a linear time course for 5 min at 30 C and pH optima at pH 7.6 and 9.0. Saturation of the enzyme was observed at 1.8 mM adenosine triphosphate (ATP) and the double reciprocal plot of activity vs. ATP concentration was nonlinear giving two apparent Km values of 0.094 and 0.518 mM. The apparent Km for glycerol, 0.112 mM, was obtained from a linear double reciprocal plot, and the enzyme was saturated at about 0.4 mM glycerol. The activity of glycerokinase in human adipose tissue excised under general anaesthesia was low and was unrelated to adipose cell size or the degree of obesity of the subject from whom the fat was obtained.


Some respiratory and metabolic effects of exercise in moderately obese men. The effects of varying levels of exercise on oxygen uptake, CO2 production, blood pressure, arterial blood gasses, and arterial concentrations of glucose, insulin, and growth hormone were examined in ten normal weight and ten moderately overweight young men. At comparable external work loads with a bicycle ergometer, the lean men required less oxygen than the obese men. When oxygen uptakes were matched during exercise on a treadmill, the lean men were walking on a steeper grade or at a higher rate than the obese men. The efficiency of exercise as assessed by the relation between oxygen uptake and work did not differ between the two groups. Blood pressure rose more in the obese during exercise than in the lean. The fall in lactate and rise in bicarbonate was of greater magnitude during cycle ergometry than during treadmill exercise. Obese and lean men, however, showed similar changes. With each level of exercise, there was a fall in arterial insulin levels, but the concentrations in the blood of overweight men always remained significantly above that of the normal men. Growth hormones tended to be higher in the normal weight men, but the differences were usually not significant, and there was no significant rise with exercise in either group until the highest levels of work were achieved. Glucose concentrations tended to be higher in the obese men, but fell to constant levels in both groups during exercise. Blood pressure rose to a greater extent in the overweight men during exercise.


Paradoxical reaction to a new minor tranquilizer. This case report demonstrates that paradoxical reactions to the benzodiazepine class of minor tranquilizer can occur, and that lorazepam, a new derivative, is not free from this effect. It also lends support to the hypothesis that such reactions may be precipitated by a frustrating stimulus, and this has clinical relevance in that such precipitants may well be similar to those situations producing anxiety for which the drug had been prescribed.


Enzyme transformations.

Enzyme transformations. The paper briefly discusses the current theories on the processes underlying the transformation of the catalytic activity of a number of enzymes. The phenomenon of transformation, which is induced by mild chemical effects acting on a protein, is of interest both from the point of view of the investigation of its molecular mechanisms and from the point of view of the study of several pathological states under which this phenomenon is observed. Gorkin's own data are presented.


Interaction of transfer RNA with 50S ribosomal subunits of Escherichia coli in absence of templates. The equilibrium constant of a complex of tRNA with the 50S ribosomal subunit was measured in the absence of a template. It was shown that the stability of the complex increases with an increase in the concentration of Mg2+, it decreases with an increase in the concentration of univalent ions, and does not depend on the pH of the medium in the range of 7.0-8.2. Removal of the 3'-terminal nucleoside of tRNA weakens the association approximately 40-fold; the subsequent successive splitting off of another three nucleotides has little effect on the association constant. In 90% 2H2O the stability of the complex increases approximately four-fold, which points to the large contribution of the hydrogen bonds to the free energy of the interaction. The tetranucleotide TphiCG competes slightly with tRNA for sites on the 50S subparticles; this means that the TphiC segment of tRNA does not play an important role in the formation of the complex under investigation.


Presence of a proteinase in polyribosomes of rat liver. The presence of a proteinase in the composition of the ribosomal proteins of rat liver is demonstrated. The enzyme possesses optimal activity in the zone of pH 7.0-7.2. Soybean trypsin inhibitor and 1-chloro-4-phenyl-3-tosylamido-2-butanone inhibit the enzyme by 50-60%.