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Biomedical subjects

S Thompson

Publications and source records attributed to S Thompson.

At least 235 records · Page 13Linked to original sources

Genetic characterization of frameshift suppressors with new decoding properties.

Suppressor mutants that cause ribosomes to shift reading frame at specific and new sequences are described. Suppressors for trpE91, the only known suppressible -1 frameshift mutant, have been isolated in Escherichia coli and in Salmonella typhimurium. E. coli hopR acts on trpE91 within the 9-base-pair sequence GGA GUG UGA, is dominant, and is located at min 52 on the chromosome. Its Salmonella homolog maps at an equivalent position and arises as a rarer class in that organism as compared with E. coli. The Salmonella suppressor, hopE, believed to be in a duplicate copy of the same gene, maps at min 17. The +1 suppressor, sufT, acts at the nonmonotonous sequence CCGU, is dominant, and maps at min 59 on the Salmonella chromosome.

Base Sequence↗

Suppression of a -1 frameshift mutation by a recessive tRNA suppressor which causes doublet decoding.

sufS was found to suppress the only known suppressible-1 frameshift mutation, trpE91, at a site identified as GGA and mapped within the single gene of the only tRNA that can decode GGA in Escherichia coli. It mapped to the same gene in Salmonella typhimurium. sufS alleles were recessive, and dominant alleles could not be isolated. This is in contrast to all other tRNA structural gene mutations identified thus far that cause frameshift suppression. The recessiveness implies that all sufS alleles are poor competitors against their wild-type tRNA(Gly2) counterparts. The base G immediately 5' of the GGA suppression site influenced the level but was not critical for suppression by sufS601. From this result, it is inferred that sufS601 causes frameshifting by doublet decoding.

Alleles↗

Uterine junctional zone: MR study of water content and relaxation properties.

Weighted samples of uterine myometrium, junctional zone, and endometrium were excised for determination of T1 and T2 in a 20-MHz spectrometer and then dried for determination of water content. The remainder of each uterus, examined with special histopathologic stains, demonstrated no significant difference in the number of blood vessels, smooth muscle cells, fibroblasts, elastin, iron, collagen, mucin, polysaccharide, or amyloid. The junctional zone water content (79.28%) was significantly lower than that of endometrium (82.88%, P = .004) and myometrium (81.05%, P = .005). The T1 of the junctional zone (ie, 643) was significantly lower than that of endometrium (ie, 836; P = .004) and myometrium (ie, 709; P = .01). The T2 of the junctional zone (ie, 58) was significantly lower than that of endometrium (ie, 87; P = .003) and myometrium (ie, 67; P = .006). The low signal intensity of the junctional zone on T2-weighted MR images is accounted for by its lower water content and T2, while the brightness on T1-weighted images is due to its reduced T1.

Adult↗

Stimulus equivalence and rule following.

The present study examined the occurrence of a novel behavior pattern with respect to a novel configuration of stimuli enabled by the participation of those stimuli in equivalence classes. In Experiment 1, functional substitutabilities were established via equivalence between two independent sets of musical stimuli. Aspects of stimuli from the two sets were then compounded to produce novel stimulus configurations. Behavioral components enabled by each separate class combined to produce novel musical performances and accurate descriptions of them. In Experiment 2, the impact of experimenter-provided names for equivalence classes on the musical performances was investigated in naive subjects by establishing similar classes without experimenter-provided names. The results indicated few differences in the playing performances under these conditions. These experiments demonstrated a possible method for the analysis of rule following.

Journal Article↗

Phytohemagglutinin skin test responses to evaluate in vivo cellular immune function in rats.

It is often necessary to have a small animal model which permits the sequential evaluation of functional immune status over a period of time. We report here the in vivo, intradermal response to phytohemagglutinin which produces an area of induration that is histologically similar to a typical delayed cutaneous hypersensitivity response, and that provides fast, quantitative, reproducible results similar to those observed with standard but more laborious and variable in vitro tests of immune function. For small animal studies this has the advantage of permitting longitudinal evaluations over time without sacrificing the animal. Using phytohemagglutinin-microprotein (0.2 mg/0.1 ml), injected intradermally, a delayed cutaneous hypersensitivity-like response is induced which is maximal at 24 hr. When immune function was altered either by treatment with a chemical immunosuppressant (ethanol) or by hormonal manipulations (hypophysectomy and rat growth hormone), the delayed cutaneous hypersensitivity-like response (area of induration) correlated closely with both macrophage migration inhibitory factor changes (r = 0.98; P less than 0.001) and mixed lymphocyte reaction changes (r = 0.99; P less than 0.05). These observations suggest that this technique correlates well with standard in vitro measures of immune response and may thus permit an in vivo estimation of immune reactivity.

Animals↗

Clustered distribution and variability in kinetics of transient K channels in molluscan neuron cell bodies.

The spatial distribution of transient K current, IA, was studied using a combination of patch-clamp and whole-cell voltage-clamp techniques. The average IA current density in somatic patches is 0.64 times the current density in the entire axotomized cell body, a finding which suggests that the axon hillock or initial segment of the axon has a higher concentration of IA channels than much of soma. The highest density of active channels during the peak IA is 1/micron2 at a membrane voltage of -20 mV. There is no evidence for a gradient in the distribution of IA channels in the cell body, but the channels are not evenly distributed. The variability in the number of channels per patch for multiple patches on the same neuron is much higher than expected for a random distribution. Statistical analysis of the data yields a coefficient of dispersion of 8.1, a value indicating a high degree of clustering. The utility of this statistic for evaluating channel distributions is discussed. Several lines of evidence suggest that the upper limit for the area of IA channel clusters is approximately 250 micron2. Single-channel currents attributed to IA were recorded in the cell-attached configuration. The voltage dependence of channel opening and inactivation are the same as measured in whole-cell voltage-clamp experiments. The single-channel conductance is about 9 pS in normal saline. Patches 9-30 micron2 in areas that contain IA channels are often devoid of other K channel types, suggesting that IA channels can occur in isochannel clusters. IA inactivation follows an exponential time course in all of the neurons examined, but the time constant of inactivation ranges from 25 to 560 msec in different cells. The voltage dependence of activation and inactivation and the reversal potential of the current are approximately the same in all cells. When multiple patches on the same neuron are studied, it is found that IA inactivates exponentially with approximately the same time constant in each patch, regardless of patch area. The data suggest that each neuron expresses predominantly, and perhaps exclusively, a single type of IA channel with distinct kinetic properties. The wide range of IA inactivation time constants observed in different cell suggests that a large number of channel types are available for expression. Possible mechanisms for generating diversity in channel types are discussed.

Animals↗

Are herbal teas safe for infants and children?

The use of herbal teas is now widespread in our community--even among children. The effect of dosing children with these products is unknown. The authors feel that the potential hazards associated with herbal teas need to be exposed.

Beverages↗

A beneficial effect of trypsin on the purification of turnip mosaic virus (TuMV) and other potyviruses.

The effects of treatment with trypsin during the purification of turnip mosaic virus (TuMV), on virus yield, infectivity and integrity of virus coat protein were examined. Trypsin increased yield markedly, and at a low concentration, increased infectivity. These effects were probably due to reduced aggregation of virus particles. At higher concentrations of trypsin, there was some degradation of virus coat protein, and infectivity was reduced. Treatment with trypsin at the optimum concentration can significantly improve purification of TuMV; more limited experiments suggest that it can also be applied to other potyviruses.

Capsid↗

Presence of antibodies to the major anaerobically induced gonococcal outer membrane protein in sera from patients with gonococcal infections.

Anaerobically grown Neisseria gonorrhoeae induces and represses the synthesis of outer membrane proteins. One of the anaerobically induced proteins, Pan 1, reacted strongly on Western blots with sera from patients with uncomplicated gonococcal infection, pelvic inflammatory disease, and disseminated gonococcal infection, but not with normal human serum. The pattern of reactivity of the sera against Pan 1 from several gonococcal strains suggested that the protein was antigenically heterogeneous, containing both common and unique epitopes. Staphylococcus aureus V8 protease digestion of Pan 1 from four gonococcal strains revealed the presence of common peptides, with one strain also containing some unique peptides and lacking others. The class of the antibody reactive with gonococcal outer membrane antigens was examined; anti-Pan 1 antibody was found to be IgG or IgM, but not IgA. The IgM antibody present reacted predominantly with Pan 1. These data indicate that the Pan 1 protein is expressed in vivo and strongly suggest that N. gonorrhoeae can grow anaerobically in vivo.

Anaerobiosis↗

Fucosylated forms of alpha-1-antitrypsin that predict unresponsiveness to chemotherapy in ovarian cancer.

We have discovered modified fucosylation of alpha 1-antitrypsin (F-AT) in the sera of ovarian cancer patients. This was detected by SDS/electrophoresis and silver-staining after extracting the sera with the fucose-binding lectin, Lotus tetragonolobus, and was identified as alpha 1-antitrypsin by Western blotting. Initially, high F-AT levels appeared to be related to the recurrence of cancer, but later measurements showed that elevated levels were also present in patients who did not respond to therapy. Using an arbitrary grading system, the level of F-AT was assessed in pairs of sera from 29 ovarian cancer patients undergoing therapy; one specimen collected just after the start of therapy and the other on a later occasion. In 75% of the 15 non-responders, F-AT was higher when measured on a second occasion; whereas in 86% of the 14 responders the second measurement was either unchanged or lower, being frequently undetectable. F-AT levels were also low or undetectable in sera from healthy women. Eight responders were monitored for F-AT throughout cyclophosphamide chemotherapy. Despite a high tumour burden at the start of therapy, all patients had relatively low levels of F-AT and this was maintained throughout remission; the levels only becoming elevated with the recurrence of tumour growth. Increased F-AT expression did not appear to be particularly associated with the presence of liver metastases and frequently predated any clinical signs of a recurrence. The interesting characteristics of these molecules could make them useful in the management of ovarian cancer.

Adult↗

Dry reagent immunoassay for measuring phenobarbital in serum and plasma.

A reagent strip has been developed for measuring phenobarbital concentrations in serum or plasma. This strip utilizes the apoenzyme reactivation immunoassay system (ARIS) and is designed for use with the Ames Seralyzer reflectance photometer. The test takes 85 s and compares strip reactivities with a two-point calibration line that is stored in the instrument. Within-run precision coefficient of variation ranges from 2.4 to 4.1% over five concentrations while the between-run precision ranges from 2.3 to 4.9%. Results obtained with clinical serum samples correlated well (r greater than 0.99) with those obtained by a fluorescence polarization immunoassay (TDx). The strips crossreact with other barbituric acid derivatives that are not coadministered with phenobarbital. Crossreactivity to the metabolite p-hydroxyphenobarbital is sufficiently low as to not interfere in the usual specimens. Samples that are hemolyzed, highly icteric, or from uremic patients should be avoided. This assay is a rapid and convenient method for monitoring serum or plasma phenobarbital concentrations. It is particularly well suited for decentralized test sites, such as emergency rooms, urgent care centers, and physician's offices.

Apoenzymes↗

The selenium status of children with phenylketonuria: results of selenium supplementation.

The selenium status of children with phenylketonuria on a synthetic low phenylalanine diet was assessed. Correlation between blood selenium and red cell glutathione peroxidase was unsatisfactory (r = 0.65) due to the poor discrimination of red cell glutathione peroxidase with a low selenium diet. No symptoms of deficiency were observed. Supplementation with 50 micrograms per week of selenium as brewers yeast tablets over a period of 6 months significantly increased the blood selenium of the phenylketonuric children. Plasma Vitamin E levels were within normal limits. The supplementation effectively doubled their selenium intake to 15-17 micrograms per day, which is probably sufficient for this group with an adequate Vitamin E status, though considerably lower than the recommended minimum intake of 50 micrograms per day.

Adolescent↗

Infant feeding in Sydney: a survey of mothers who bottle feed.

Two hundred and seventy-four mothers who were bottle feeding their infants aged from 1 to 9 months, were interviewed. Particular attention was paid to mixing technique, storage of reconstituted formulae, the reasons for deciding to bottle feed and what influenced the choosing of a particular milk. Following the interview, a sample of milk from a previously prepared bottle was taken to measure osmolality and bacterial colony count. Eighty-two per cent of the mothers had initially attempted breast feeding. The commonest reason for changing to a bottle was that the mother felt she was not able to produce enough milk to satisfy the baby. Thirty per cent chose the milk they were using on the recommendation of the maternity hospital. Errors in reconstituting the formulae, compared with the manufacturer's instructions were made by 100 (30%) mothers. In 52 cases these were potentially serious errors, usually erring on the side of preparing an over-concentrated formula. This finding from the interviews was confirmed by osmolality analysis of milk samples. Twenty-two per cent of samples collected grew potential pathogens. There was no correlation between the presence of bacterial growth and the use of prewarmed formulae stored in insulated carriers. No correlation was found between triceps skinfold thickness, age of introduction of solids or method of reconstitution of formula.

Australia↗

Inward rectification in response to FMRFamide in Aplysia neuron L2: summation with transient K current.

The response of Aplysia abdominal ganglion neuron L2 to the molluscan neuroactive peptide Phe-Met-Arg-Phe-NH2 (FMRFamide) was studied in voltage-clamp experiments. In all of the experiments, focal application of the peptide to the soma activated an inward rectifier current and reduced the apparent amplitude of the transient K current, IA. In a few cells, Na and K currents were activated in addition to these effects. Voltage-jump experiments were performed to study the ionic dependence, kinetics, and voltage dependence of the inward rectifier. Inward rectification increased exponentially during hyperpolarizing pulses and recovered exponentially on return to the resting potential. The reversal potential was variable, but was near -40 mV at the beginning of experiments. Inward rectification was insensitive to changes in external Na, Ca, or K concentration, but lowering the external Cl concentration had complicated effects on current amplitude. When KCl microelectrodes were used, perfusion with low-Cl external saline increased the amplitude of the peptide-dependent inward rectifier and shifted its reversal potential to a more positive voltage. With KAc microelectrodes, perfusion with low-Cl saline reduced the amplitude of the current. Inward rectification increased when a KAc microelectrode was withdrawn and replaced with a low-resistance KCl electrode, even when there was no measurable change in reversal potential. These results suggest that the FMRFamide-dependent inward rectifier is a Cl current that, like the current described by Chesnoy-Marchais (1982, 1983), is modulated by intracellular Cl. FMRFamide reduced the apparent amplitude of IA without affecting the voltage dependence of IA activation or inactivation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Spatial distribution of Ca currents in molluscan neuron cell bodies and regional differences in the strength of inactivation.

The spatial distribution of Ca current in molluscan neuron cell bodies was studied using a large patch method in combination with 2-microelectrode voltage clamp. The method has a spatial resolution equal to about 0.1% of the cell body area. Ca current is not uniformly distributed. The current density varies between patches, changing by as much as a factor of 2.5 over a distance of 20 micron, and there is evidence that Ca current occurs in "hot spots" involving a few hundred channels. The current density increases in a moderately steep gradient from the soma cap, opposite the axon, toward the axon hillock. Ca currents in patches from different regions of the soma are qualitatively different. Currents near the soma cap do not inactivate or inactivate weakly during depolarization, while currents of equal density nearer the axon hillock exhibit pronounced inactivation. The strength of inactivation increases in parallel with the gradient in current density, but local differences in current density, or in the number of active Ca channels, do not explain the variability in inactivation. Inactivating and noninactivating Ca currents could not be distinguished on the basis of activation or deactivation kinetics, voltage dependence of activation, or sensitivity to hyperpolarizing conditioning pulses. Also, the amplitude of noninactivating current near the soma cap is reduced by intracellular Ca injection showing that, like the whole-cell current, Ca current in this region is subject to Ca-dependent inactivation. The data favor the hypothesis that these cells express only one type of Ca channel. Differences in the strength of inactivation may result from local differences in cytoplasmic Ca buffering, local modification of Ca channels in a way that changes their sensitivity to Ca-dependent inactivation, or local differences in the availability of cytoplasmic factors or enzymes that are necessary for inactivation.

Animals↗

Mutants of elongation factor Tu promote ribosomal frameshifting and nonsense readthrough.

This is the first report of ribosomal frameshifting promoted by mutants of the elongation factor Tu (EF-Tu). EF-Tu mutants can suppress both -1 and +1 frameshift mutations. The level of nonsense readthrough is also increased at some UGA (this paper) and UAG (Hughes, 1987) sites by these mutants. Suppression occurs when a mutant tuf allele is paired with a wild-type copy of the other tuf gene but is most efficient when both tuf genes are mutant. Frameshifting mediated by the tuf alleles studied, tufA8 and tufB103, is not general; indeed most frameshift mutations are not suppressed. Several possible mechanisms by which mutant EF-Tu may cause frameshifting are discussed.

Alleles↗