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Biomedical subjects

S Thompson

Publications and source records attributed to S Thompson.

At least 271 records · Page 15Linked to original sources

Analysis of FMRF-amide effects on Aplysia bursting neurons.

The peptide L-phenylalanyl-L-methionyl-L-arginyl-L-phenylalaninamide (FMRF-amide) was pressure-applied onto the somata of bursting neurons L4 and L6 in the Aplysia abdominal ganglion. FMRF-amide causes a biphasic response, first depolarizing and then hyperpolarizing the neuron. In voltage-clamp experiments, FMRF-amide induces an inward current that begins 100-200 msec after applying the peptide and peaks in 2-10 sec. This is followed by an outward current that begins with a latency of 2-5 sec and peaks in 15-65 sec. The entire response lasts 1-5 min. Experiments were done to separate the two currents induced by FMRF-amide on the basis of ion selectivity and kinetics and to determine their I(V) relationships. The currents were studied using a method to quickly measure I(V) curves. The inward current is caused by a conductance increase and has a reversal potential of approximately +18 mV. This current depends on the concentration of extracellular Na ions but not Ca, Cl, or K ions and is insensitive to tetrodotoxin, hexamethonium, and curare. The outward current is caused by a conductance increase and has a reversal potential of approximately -61 mV, which is similar to the reversal potential of the fast, transient K current (IA) in the same cells. This current is sensitive to changes in the external K ion concentration but not to changes in Cl, Ca, or Na concentration. The outward current is partially blocked by 1 mM 4-aminopyridine but not TEA or curare. Neither current is significantly voltage dependent within the range from -70 to -40 mV.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Slow outward tail currents in molluscan bursting pacemaker neurons: two components differing in temperature sensitivity.

Bursting pacemaker neurons of Tritonia and Aplysia were studied in voltage-clamp experiments to determine the ionic requirements of the slow outward tail current. This current is important for terminating bursts and hyperpolarizing the neuron during the interval between bursts and therefore contributes to the timing of pacemaker activity. A method for rapid extracellular perfusion of the neuron soma with solutions of different ionic composition was used to study the ionic dependence of the tail current. Measurements of the current-voltage relationship were made at different times during the decay of the tail current to determine the reversal potential in different ionic solutions. These experiments showed that 2 or more slow outward currents contribute to the tail current. Temperature had a large effect on these currents. At cold temperatures (10-15 degrees C), the tail current was predominantly a K current. At warm temperatures (20-23 degrees C), both a K current and a K-insensitive outward current were seen. Bursting pacemaker activity occurred throughout this temperature range. Both the K current and the K-insensitive current required Ca influx for activation. These findings help to reconcile conflicting reports in the literature concerning the ionic dependence of the slow outward tail current and suggest a mechanism for temperature compensation of bursting activity in these pacemaker cells.

Action Potentials↗

Ethoxy-, pentoxy- and benzyloxyphenoxazones and homologues: a series of substrates to distinguish between different induced cytochromes P-450.

The individual members of a homologous series of phenoxazone ethers related to ethoxyresorufin were O-dealkylated, and the parent compound phenoxazone was ring-hydroxylated, each at different rates with hepatic microsomes of untreated rats. A structure-activity relationship (SAR) was plotted, relating the rate of O-dealkylation to the length and type of the ether side-chain. Phenobarbitone (PB), 3-methylcholanthrene (MC), Aroclor 1254 (ARO), isosafrole (ISO) and SKF-525A each induced preferentially the O-dealkylation of different members of the homologous series, resulting in the appearance of 5 different SAR plots, which characterized and differentiated between the 5 different inducers. beta-Napthoflavone (BNF) had a similar effect to MC, whereas pregnenolone 16 alpha-carbonitrile treatment caused no large change in the metabolism of any of the substrates tested. For characterizing the effects of the different inducers it was largely sufficient to compare the O-dealkylations of just 4 of the ethers: methoxy-, ethoxy-, pentoxy- and benzyloxyphenoxazone. Very high degrees of induction were seen. MC and ARO each induced preferentially the O-dealkylation of ethoxyphenoxazone (51- and 61-fold respectively). PB and SKF-525A each induced preferentially the O-dealkylation of pentoxyphenoxazone (283- and 324-fold respectively). ISO induced preferentially the O-dealkylation of benzyloxyphenoxazone (43-fold). For any particular induced type of microsomes the substrate with the fastest metabolism was not necessarily the substrate whose metabolism was induced the most, so that in order to characterize each of the 5 different inducers (PB, MC/BNF, ARO, ISO, SKF) it was necessary to compare both the degrees of induction and the specific activities of the reactions. Experiments with purified cyt. P-450 isozymes showed that ethoxyphenoxazone and pentoxyphenoxazone were highly selective substrates for the major isozymes induced by MC and PB respectively, whilst benzyloxyphenoxazone was a good substrate for both isozymes. Experiments using the organic inhibitors metyrapone and alpha-naphthoflavone and inhibitory antibodies against individual cyt. P-450 isozymes indicated that similar substrate selectivities occurred with the monooxygenase system in the microsomal membrane. It is suggested that the use of some or all of these homologous phenoxazone ethers will provide both a simple routine test for the characterization of several types of inducing agents and a powerful tool for investigating the biochemical basis for cyt. P-450 isozyme substrate selectivity.

Animals↗

Measurement of gastric emptying rates by radioactive isotope scanning and epigastric impedance.

When liquids of low electrical conductivity enter the stomach the impedance of the epigastric region to a 4 mA, 100 kHz current increases. There follows a decline, which logically represents gastric emptying. This method of measuring gastric emptying was compared against scintigraphy in six volunteers, and similar results were obtained. Impedance monitoring is entirely noninvasive, inexpensive, simple, and quick. The method merits further exploration.

Adult↗

Measurement of the diene conjugated form of linoleic acid in plasma by high performance liquid chromatography: a questionable non-invasive assay of free radical activity?

It has been previously reported that the main diene-conjugated fatty acid in human plasma is a non-oxygen containing linoleic acid isomer (PL-9, 11-LA'). It has also been proposed that this isomer can be used as a specific marker of free radical-mediated lipid peroxidation in humans. Here we report that the in vitro induction of lipid peroxidation in human and rat blood with either UV irradiation or phenylhydrazine failed to increase the plasma levels of this isomer. The induction of lipid peroxidation in vivo in rats pretreated with either phenylhydrazine or bromotrichloromethane also failed to increase the plasma levels of this isomer. These findings demonstrate that PL-9, 11-LA' cannot be used as an in vivo marker of free radical-mediated lipid peroxidation in rats and casts doubts on its validity as a specific marker in humans.

Animals↗

Treating gonococcal infections resistant to penicillin in Bangkok: comparison of cefuroxime and spectinomycin.

Gonococcal organisms have become resistant to antimicrobials throughout the world. Such resistance is common in Thailand, where 40% of gonococci produce penicillinase (PPNG strains) and over half the remainder have MICs of penicillin greater than or equal to 1 mg/l. To evaluate the effectiveness of cefuroxime against such resistant organisms, a controlled clinical trial comparing spectinomycin and cefuroxime was conducted at Bangrak Hospital, Bangkok, in 1982-3. Of 472 patients who were randomly assigned to treatment, 365 (77%) yielded positive cultures before treatment and returned for follow up evaluation three to 13 days after treatment. Of the 365 patients, 359 (98%) were cured, and no difference between the two treatment regimens was found either by the sex of the patient or by the presence of PPNG strains. The MIC of cefuroxime against all organisms was less than or equal to 1 mg/l. In vitro susceptibilities of gonococci in Bangkok have not changed appreciably during the past two years. Regimens of cefuroxime and spectinomycin are highly effective even for the relatively resistant gonococci in Bangkok. The pharmacokinetics, in vitro susceptibilities, and effectiveness of cefuroxime encourage evaluation of lower doses of the drug.

Adolescent↗

Synthesis of apolipoproteins, alphafoetoprotein, albumin, and transferrin by the human foetal yolk sack and other foetal organs.

Fragments of human foetal organs and blood at 5-11 weeks of postfertilization development were cultured in radioactive protein precursors. The secreted products were characterized by immunoprecipitation, and by measuring the mobility of the immunoprecipitates on polyacrylamide gels. It was found that secondary human yolk sacks secreted apolipoproteins A1 and B. The work of previous authors on the synthesis of other serum products by this organ and by the foetal liver and by the foetal intestines was confirmed. Within the yolk sack, the endoderm, the blood cells, and the outside epithelium reacted with antibodies against apolipoprotein A1 and transferrin. By metabolic labelling of umbilical cord blood, it was found that blood did not secrete apolipoproteins A1 and B. Blood cells could therefore not be a source of these secreted products.

Albumins↗

A monoclonal antibody which detects a 125 kd glycoprotein on embryonal carcinoma cells and is mitogenic for murine spleen cells.

The isolation and characterisation of a monoclonal antibody (2H9), selected for reactivity against murine teratocarcinoma stem cells, is described. This antibody recognizes one major glycoprotein on the surface of embryonal carcinoma (EC) cells but detects multiple glycoprotein species on various murine or human derived cell types. The binding of the antibody to cells is abolished by treatment with periodate or neuraminidase. These results suggest that 2H9 antibody detects a carbohydrate determinant; this is apparently distinct from other anti-EC reactive monoclonal antibodies and a variety of lectins. These antibodies are mitogenic for a population of lymphocytes in murine spleen. The antibodies recognize numerous cell surface glycoproteins on the surface of the spleen cells.

Animals↗

Rapid recovery from K current inactivation on membrane hyperpolarization in molluscan neurons.

Recovery from K current inactivation was studied in molluscan neurons using two-microelectrode and internal perfusion voltage clamps. Experiments were designed to study the voltage-dependent delayed outward current (IK) without contamination from other K currents. The amount of recovery from inactivation and the rate of recovery increase dramatically when the membrane potential is made more negative. The time course of recovery at the resting potential, -40 mV, is well fit by a single exponential with a time constant of 24.5 s (n = 7). At more negative voltages, the time course is best fit by the sum of two exponentials with time constants at -90 mV of 1.7 and 9.8 s (n = 7). In unclamped cells, a short hyperpolarization can cause rapid recovery from inactivation that results in a shortening of the action potential duration. We conclude that there are two inactivated states of the channel and that the time constants for recovery from both states are voltage dependent. The results are discussed in terms of the multistate model for K channel gating that was developed by R. N. Aldrich (1981, Biophys. J., 36:519-532).

Action Potentials↗

Cloned human teratoma cells differentiate into neuron-like cells and other cell types in retinoic acid.

Single cell clones were isolated from the human teratoma line, Tera-2. The cells of three of these clones were studied. The progressively growing cells were shown to be tumorigenic, and they were characterized by the lack of expression of beta 2-microglobulin and HLA-A,B,C determinants on the cell surface. The majority of the cells expressed Thy-1 antigen and a 90 X 10(3) molecular weight protein recognized by the monoclonal antibody F10.44.2; between a third and half of the cells expressed the sugar specificities detected by the anti-SSEA-1 monoclonal antibody. In response to 5 X 10(-5) M-retinoic acid applied to cells in monolayer culture, the cells differentiated into a population of flat static cells arrested in the G1 phase of the cell cycle. A substantial proportion of these differentiated cells expressed beta 2-microglobulin and 43 X 10(3) molecular weight HLA-A,B,C polypeptides, Thy-1, SSEA-1 sugar determinants, and the 90 X 10(3) Mr protein recognized by F10.44.2. The apparent molecular weight of fibronectin secreted by the cells decreased by about 5 X 10(3) Mr to 235 X 10(3) Mr after differentiation. The progressively growing cells lacked reactivity with reagents that mark cells in the nervous system. Following aggregation and retinoic acid treatment, neuron-like cells were formed. These cells reacted with reagents that also react with human neurons in culture: they reacted with tetanus toxin, the anti-neurofilament antibodies BF10 and RT97, the anti-ganglioside, GQ1c antibody F12 A2B5, and anti-Thy-1. The progressively growing cells of these Tera-2 clones are therefore capable of forming at least two types of cell: the flat cells in monolayer cultures and the neuron-like cells. None of the cell populations reacted with the monoclonal antibody against SSEA-3 and these cloned cells are therefore distinct from previous isolates from Tera-2.

Animals↗

Facilitating visual discrimination learning of moderately and severely mentally retarded children through illumination of stimuli.

The effectiveness of illuminating the positive stimulus after a correct response in visual discrimination training of moderately and severely mentally retarded children was investigated. An economical unit for illumination of stimuli was designed for purposes of the present investigation and for possible use in nonspeech communication training of mentally retarded children. In Experiment 1, the illumination of black stimuli on white backgrounds failed to facilitate discrimination learning. In Experiment 2, however, illumination of white stimuli on black backgrounds facilitated discrimination learning. The findings in the second experiment support the use of changes in illumination as an attention-influencing feature of nonspeech communication keyboards for retarded children.

Attention↗