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Biomedical subjects

T Palmer

Publications and source records attributed to T Palmer.

At least 55 records · Page 3Linked to original sources

Local-level and global-level form characteristics in apparent-motion correspondence.

This study addressed the "correspondence" problem of apparent-motion (AM) perception in which parts of a scene must be matched with counterparts separated in time and space. Given evidence that AM correspondence can be mediated by two distinct processes--one based on a low-level motion-detection mechanism (the Reichardt process), the other involving the tracking of objects by visual attention (the attention-based process)--the present study explored how these processes interact in the perception of apparent motion between hierarchically structured figures. In three experiments, hierarchical figures were presented in a competition motion display so that, across frames, figures were identical at either the local or the global level. In experiment 1 it was shown that AM occurred between locally identical figures. Furthermore, with the Reichardt AM component eliminated in experiments 3 and 4, no preference was obtained for either level. While evidence from previous studies that form extraction for hierarchically structured figures proceeds from the global to the local levels, the present results indicate the irrelevance of such a global precedence in AM correspondence. In addition, it is suggested that Reichardt AM correspondence between local elements constrains attention-based AM correspondence between global figures so that both components move in the same direction. It is argued that this constraining process represents an elegant means of achieving AM correspondence between objects undergoing complex transformation.

Adult↗

The mob locus of Escherichia coli K12 required for molybdenum cofactor biosynthesis is expressed at very low levels.

The mob locus of Escherichia coli encodes functions which catalyse the synthesis of active molybdenum cofactor, molybdopterin guanine dinucleotide, from molybdopterin and GTP. Reporter translational lac fusion mutations in the mobA gene have been constructed using lambda placMu9 mutagenesis. The mob locus is expressed at very low levels under both aerobic and anaerobic growth conditions. Neither additions to the growth media (nitrate, tungstate or molybdate) nor secondary mutations at the moa, mob, mod, moe or mog loci affected the level of expression. Two transcription initiation sites and their associated promoter regions have been identified upstream of mobA. Both of the promoter regions show a poor match to the -35 and -10 consensus sequences for sigma 70 promoters. A 2.2 kb chromosomal DNA fragment which complemented all available mob mutants has been sequenced. Two ORFs were identified, arranged as a single transcription unit. The encoded polypeptides have predicted molecular masses of 21642 Da and 19362 Da, respectively. The DNA has been subcloned into a T7 overexpression system and the predicted products identified. The mobA gene encodes protein FA, which has been purified to homogeneity and brings about the activation of inactive molybdoenzymes in cell extracts of mob mutants. The mobB gene encodes a polypeptide with a putative nucleotide binding site. All available mob mutations which have been selected for by their ability to grow anaerobically in the presence of chlorate are located in the mobA gene.

Amino Acid Sequence↗

Purification of a Candida albicans germ tube specific antigen.

In a previous work, Marot-Leblond et al. identified a Candida albicans germ tube-specific antigen by the use of a monoclonal antibody (mAb 3D9.3). In the present report, we used a two-step procedure to obtain a purified preparation of this antigen from a Zymolyase extract of Candida albicans germ tubes. The extract was first fractionated by gel filtration chromatography. The immunoreactive fractions were pooled, and the 3D9.3 antigen was further purified by hydrophobic interaction chromatography using a Phenyl-superose column. Analysis by SDS-PAGE, immunoblotting and Concanavalin A staining, revealed a single, polydisperse band ranging from 110 to 170 kDa. The antigen was purified 126-fold by protein content and 16.4-fold by carbohydrate content. Recovery of the antigen was 6.8% following the two-step purification.

Animals↗

Isolation of protein FA, a product of the mob locus required for molybdenum cofactor biosynthesis in Escherichia coli.

The mob mutants in Escherichia coli are pleiotropically defective in all molybdoenzyme activities. They synthesise molybdopterin, the unique core of the molybdenum cofactor, but are unable to attach the GMP moiety to molybdopterin to form molybdopterin guanine dinucleotide, the functional molybdenum cofactor in Escherichia coli. A partially purified preparation termed protein FA (protein factor d'association), is able to restore molybdoenzyme activities to broken cell preparations of mob mutants. A fragment of DNA capable of complementing mob mutants has been isolated from an E. coli genomic library. Strains carrying this DNA in a multicopy plasmid, express 30-fold more protein FA activity than the wild-type bacterium. Protein FA has been purified to homogeneity by a combination of ion-exchange, affinity and gel-filtration chromatography. Protein FA consists of a single polypeptide of molecular mass 22 kDa and is monomeric in solution. N-terminal amino acid sequencing confirmed that protein FA is a product of the first gene at the mob locus. The purified protein FA was required in stoichiometric rather than catalytic amounts in the process that leads to the activation of the precursor of the molybdoenzyme nitrate reductase, which is consistent with the requirement of a further component in the activation.

Amino Acid Sequence↗

Inhibition of proton-translocating transhydrogenase from photosynthetic bacteria by N,N'-dicyclohexylcarbodiimide.

The effects of N,N'-dicyclohexylcarbodiimide [(cHxN)2C] on the proton-translocating enzyme, NAD(P) H(+)-transhydrogenase (H(+)-Thase), from two species of phototrophic bacteria have been investigated. The polypeptides of H(+)-Thase from Rhodobacter capsulatus are membrane-associated, requiring detergent to maintain solubility. The enzyme from Rhodospirillum rubrum, however, has a water soluble polypeptide (Ths) and a membrane-associated component (Thm) which, separately, have no activity but which can be fully reconstituted to give a functional complex. Two observations suggest that (cHxN)2C inhibited H(+)-Thase from both species by modification either close to or at the NADP(H)-binding site on the enzyme: (a) the presence of NADP+ or NADPH caused increased inhibition by (cHxN)2C and (b) after treatment of the purified enzyme from Rb. capsulatus with (cHxN)2C, the release of NADP+ became rate-limiting, as evidenced by a stimulated rate of NADPH-dependent reduction of acetylpyridine adenine dinucleotide by NADH. Experiments in which Ths and Thm from R. rubrum were separately treated with (cHxN)2C then reconstituted with the complementary, untreated component revealed that the NADP(H)-enhanced modification by (cHxN)2C was confined to Thm. In contrast to some experiments with mitochondrial H(+)-Thase [Wakabayashi, S. & Hatefi, Y. (1987) Biochem. Int. 15, 667-675], there was no protective effect of either NAD+ or NADH on the inhibition by (cHxN)2C of enzyme from photosynthetic bacteria. However, amino acid sequence analysis of proteolytic fragments of Ths revealed that the NAD(H)-protectable, (cHxN)2C-reactive glutamate residue in mitochondrial H(+)-Thase might be replaced by glutamine in R. rubrum.

Amino Acid Sequence↗

The relation between the soluble factor associated with H(+)-transhydrogenase of Rhodospirillum rubrum and the enzyme from mitochondria and Escherichia coli.

Although in mitochondria, Escherichia coli and Rhodobacter capsulatus the H(+)-transhydrogenases are intrinsic membrane proteins, in Rhodospirillum rubrum a water-soluble component (Ths) and a membrane-bound component are together required for activity. Ths was selectively removed from chromatophore membranes of Rhs. rubrum and was purified to homogeneity by precipitation with (NH4)2SO4 and ion-exchange, affinity dye and gel exclusion chromatography. The latter indicated an Mr of approx. 74,000 under non-denaturing conditions but analysis of the pure protein by SDS-PAGE revealed a single polypeptide, Mr 43,000. Antibodies against this polypeptide inhibited transhydrogenase activity of chromatophores and decreased the capacity of Ths to restore activity to depleted membranes. They reacted with a polypeptide of Mr 43,000 in crude cell extract, chromatophore membranes and chromatophore washings but not with transhydrogenase polypeptides from the membranes of E. coli, Rb. capsulatus or animal mitochondria. The N-terminal amino acid sequence of the 43,000 polypeptide was strongly homologous with the reported N-terminal regions of mitochondrial transhydrogenase and the alpha subunit of the E. coli protein. The break between the alpha and beta polypeptides of E. coli transhydrogenase is such that both components are membrane-associated. In contrast, these results suggest that in the Rhs. rubrum enzyme Ths has been formed by a break closer to the N-terminus, thus avoiding the putative trans-membrane helical segments and yielding a relatively hydrophilic subunit, which is water-soluble. There is a predicted similarity between Ths and the reported sequence of alanine dehydrogenase from Bacillus but Ths did not have any alanine dehydrogenase activity.

Amino Acid Sequence↗

Nicotinamide nucleotide transhydrogenase from Rhodobacter capsulatus; the H+/H- ratio and the activation state of the enzyme during reduction of acetyl pyridine adenine dinucleotide.

Chromatophores from Rhodobacter capsulatus were incubated in the dark with NADPH and acetylpyridineadenine dinucleotide (AcPdAD+) in the presence of different concentrations of myxothiazol. The transhydrogenase activity was monitored until an appropriate mass action ratio, [AcPdAD+][NADPH]/[AcPdADH][NADP+], was reached. The sample was then illuminated and the initial rate of either AcPdAD+ reduction by NADPH or AcPdADH oxidation by NADP+ was recorded. The ratio of H+ translocated per H- equivalent transferred by transhydrogenase was calculated from the value of the membrane potential (delta pH = 0) at which illumination caused no net reaction in either direction. The mean value for the H+/H- ratio was 0.55. At greater values of [AcPdAD+][NADPH]/[AcPdADH][NADP+] than were employed in the above experiments and over a wider range of concentrations of myxothiazol, it was found that incremental increases in the membrane potential always gave rise to a decrease, never an increase in the rate of AcPdAD+ reduction. In contrast to the H(+)-ATP synthase, there is no evidence of any activation/deactivation of H(+)-transhydrogenase by the protonmotive force.

Membrane Potentials↗

The detection of human papillomaviruses in cervical biopsies by immunohistochemistry and in situ hybridization.

The presence of human papillomavirus (HPV) types 6, 16 and 18 in cervical biopsies can be detected by an immunoperoxidase technique using type-restricted monoclonal antibodies raised against fusion proteins representing the L1 major capsid proteins of these three HPV types. In a retrospective study (n = 54) we have used these antibodies and biotinylated DNA probes of HPV 6, 16 and 18 to detect and type HPV in formalin-fixed material from the cervix. The biopsies were classified histologically into normals, wart infections without dysplasia, cervical intraepithelial neoplasia (CIN) and squamous cell carcinomas. Antibody staining showed that 22% of all CIN was positive for HPV 16 and 40% of cervical warts were positive for HPV 6, 16 and 18. There was no HPV capsid protein detected in the normals and squamous cell carcinomas using these antibodies, whereas 25% of the tumours were positive for HPV 16 by in situ hybridization. Sections of cervical warts and CIN positive for HPV types by in situ hybridization were also positive by antibody staining which suggests that both techniques are detecting replicating virus. We feel these two techniques complement each other in detection and typing of HPV in cervical biopsies from patients with active disease.

Biopsy↗

Alterations in G-protein expression and the hormonal regulation of adenylate cyclase in the adipocytes of obese (fa/fa) Zucker rats.

Attenuated maximal activations by forskolin, Mn+. NaF or guanosine 5'-[gamma-thio]triphosphate (GTP[S]) were noted for adenylate cyclase activity in adipocytes from obese (fa/fa) Zucker rats compared with their lean (Fa/Fa) littermates. GTP[S] achieved half-maximal activation of adenylate cyclase at some 10-fold lower concentrations in membranes from lean animals compared with those from obese. Levels of the 42 and 45 kDa forms of Gs were some 40-50% lower in membranes from obese animals, and levels of Gi-1 and Gi-3 were some 62-65% lower. No differences in levels of Gi-2 alpha-subunits or G-protein beta-subunits were observed. Gi function, as assessed by inhibiting forskolin-stimulated adenylate cyclase, achieved by prostaglandin E1, nicotinate and phenylisopropyladenosine, was similar in membranes from both lean and obese animals. Levels of beta-adrenoceptors were some 50% lower in membranes from obese animals. It is suggested that the attenuated activation of adenylate cyclase by stimulatory ligands in membranes from obese animals may be caused by decreases in both Gs and receptors, and that this may contribute to the attenuated lipolytic response seen in adipocytes from such animals.

Adenylyl Cyclases↗

Purification and properties of the H(+)-nicotinamide nucleotide transhydrogenase from Rhodobacter capsulatus.

1. H(+)-transhydrogenase from Rhodobacter capsulatus is an integral membrane protein which, unlike the enzyme from Rhodospirillum rubrum, does not require the presence of a water-soluble component for activity. 2. The enzyme from Rb. capsulatus was solubilised in Triton X-100 and subjected to ion-exchange, hydroxyapatite and then gel-exclusion column chromatography. SDS/PAGE of the purified enzyme revealed the presence of two polypeptides with apparent Mr 53,000 and 48,000. Other minor components which were stained on the electrophoresis gels or which were revealed on Western blots exposed to antibodies raised to total membrane proteins, were probably contaminants. 3. Antibodies raised to the 53-kDa and 48-kDa polypeptides cross-reacted with equivalent polypeptides in Western blots of solubilised membranes from Rb. capsulatus, Rhodobacter sphaeroides and Rhs. rubrum. The significance of this finding is discussed in the context of the hypothesis [Fisher, R.R. & Earle, S.R. (1982) The pyridine nucleotide coenzymes, pp. 279-324, Academic Press, New York] that the soluble component associated with H(+)-transhydrogenase from Rhs. rubrum is an integral part of the catalytic machinery. Antibodies against the 48-kDa and 53-kDa polypeptides of the Rb. capsulatus enzyme cross-reacted with equivalent polypeptides in solubilised membranes of Escherichia coli. 4. The dependence of the rate of H- transfer by purified H(+)-transhydrogenase on the nucleotide substrate concentrations under steady-state conditions, the effects of inhibition by nucleotide products and the inhibition by 2'-AMP and by 5'-AMP suggest that the reaction proceeds by the random addition of substrates to the enzyme with the formation of a ternary complex. 5. In conflict with this conclusion, the reduction of acetylpyridine adenine dinucleotide (AcPdAD+) by NADH in the absence of NADP+ by bacterial membranes was earlier taken as evidence for the existence of a reduced enzyme intermediate [Fisher, R.R. & Earle, S.R. (1982) The pyridine nucleotide coenzymes, pp. 279-324, Academic Press, New York]. However, it is shown here that although chromatophore membranes of Rb. capsulatus catalysed the reduction of AcPdAD+ by NADH, the reaction was not associated with the purified H(+)-transhydrogenase. Moreover, in contrast with the true transhydrogenase reaction, the reconstitution of AcPdAD+ reduction by NADH (in the absence of NADP+) in washed membranes of Rhs. rubrum with partially purified transhydrogenase factor, was only additive.

Bacterial Chromatophores↗

Genetically acquired diabetes: adipocyte guanine nucleotide regulatory protein expression and adenylate cyclase regulation.

Adipocyte membranes from diabetic (db/db) animals showed marked elevations in the levels of alpha-subunits for Gi-1 which were almost twice those found in membranes from their normal, lean littermates. In contrast, no apparent differences were noted for levels of the alpha-subunits of Gi-2 and Gi-3, the 42 and 45 kDa forms of Gs and for G-protein beta-subunits. Adenylate cyclase specific activity was similar in membranes from both normal and diabetic animals under basal conditions and also when stimulated by optimal concentrations of either NaF or forskolin. In contrast, the ability of isoprenaline, glucagon and secretin to stimulate adenylate cyclase activity was greater in membranes from normal animals compared with membranes from diabetic animals. Receptor-mediated inhibition of adenylate cyclase, as assessed using PGE1 and nicotinate, was similar using membranes from both sources, but PIA (phenylisopropyladenosine) was a slightly more effective inhibitor in membranes from diabetic animals. A doubling in the expression of Gi-1 thus appears to have little discernible effect upon the inhibitory regulation of adenylate cyclase.

Adenylyl Cyclases↗

Metastatic carcinoma in the neck: a clinical, radiological, scintigraphic and pathological study.

This study was undertaken to compare clinical evaluation of the neck with 99mTc(v) DMSA planar scintigraphy and computerized tomography (CT) in patients with head and neck carcinoma. Twenty-six patients were studied and in all but one the neck was previously untreated. A total of 31 neck dissections were performed and the specimens examined histopathologically. CT was approximately as accurate (71%) as clinical examination (68%) and more accurate than 99mTc(v) DMSA planar scintigraphy (48%) in predicting which necks contained metastatic carcinoma. Overall, 13% of necks had their staging correctly changed by 99mTc(v) DMSA scintigraphy compared with 10% for CT. Although scintigraphy upstaged 13% of clinically N0 necks compared to 6% for CT, it was less sensitive and specific than either clinical examination or CT. 99mTc(v) DMSA planar scintigraphy has no role to play in the investigation of patients with metastatic carcinoma to include the clinically N0 neck.

Adenocarcinoma↗

A rapid burst preceding the steady-state rate of H(+)-transhydrogenase during illumination of chromatophores of Rhodobacter capsulatus. Implications for the mechanism of interaction between protonmotive force and enzyme.

At the onset of illumination of chromatophores there was a burst (t1/2 approx. 5 ms) in the rate of the H(+)-transhydrogenase reaction before establishment of the steady-state rate. The burst was suppressed at high pH with a pKa of approx. 8.5. The burst and the steady-state rate were inhibited by either (i) a combination of myxothiazol and carbonylcyanide-p-trifluoromethoxyphenylhydrazone, or (ii) NAD+, or (iii) dicyclohexylcarbodiimide. The results support a model in which substrate binding to H(+)-transhydrogenase is relatively fast. A subsequent slow step is accelerated by the protonmotive force and a third step, possibly product release, is rate-limiting in steady-state turnover during illumination.

Hydrogen-Ion Concentration↗

Cerebral asymmetry in visual attention.

The traditional view of cerebral lateralization of various cognitive functions has been challenged by results from recent experimental and clinical studies. Evidence has been gathered to suggest that a seemingly unitized cognitive function can be further broken down into various processing subcomponents which are distributed across the two hemispheres. For instance, according to such a more complex conceptualization of cerebral lateralization, language is seen not as a unitary ability, but rather as a collection of syntactic, semantic, and prosodic components, with each lateralized in particular manners. In much the same way, the present study attempts to examine the cerebral lateralization patterns of the seemingly unitary visual perception process. In a visual half-field experiment, 20 normal subjects were asked to make same/different judgments to laterally presented arrays of stimuli of the same type as previously studied by Treisman and her colleagues in experiments attempting to separate the preattentive and attentive stages of visual perception. Hemispheric differences were obtained only in tasks requiring attentive processing (i.e., Treisman's glueing). Results indicate a local attentional strategy for the left hemisphere and a global attentional strategy for the right hemisphere.

Adult↗

Thermal effects of stationary "hot tip" laser coronary probes: an in vitro assessment.

Percutaneous laser thermal probe angioplasty requires sufficient laser probe flexibility to access the coronary tree. This may entail a loss of axial strength and the resultant slow advancement may lead to unwanted heating of the normal coronary artery proximal to the lesion. To assess the lateral thermal effects of stationary coronary laser probes, laser thermal energy (50-150 J) was delivered to 25 coronary artery segments (diameter 1.9-4.0 mm) in a perfused cadaver heart preparation using a 1.7 mm tip probe. Adherence to the vessel wall occurred in 19 segments, endothelial charring in 8 segments, and perforation in 3 segments. Endothelial charring was seen in 8 of 13 nonperfused segments but in 0 of 12 segments perfused at 60 ml/minute (P less than 0.01). In all three perforations the vessel to probe diameter ratio was less than 1.6:1, perfusion was absent, and traction to dislodge the adherent probe was necessary. Lateral wall damage is a complication of stationary laser probes: smaller-tipped probes which are advanced rapidly at the time of energy delivery may enhance the safety margins of coronary laser thermal probe angioplasty.

Angioplasty, Balloon↗

Ultrastructural morphometric analysis of papillary neoplasms: biological and diagnostic relevance.

Ten papillary adenocarcinomas of thyroid origin (P-Thy), ten papillary adenocarcinomas of ovarian origin (P-Ov), and eight papillary neoplasms of non-thyroid/non-ovarian origin (P-Other) were morphometrically compared using 19 distinct quantitative nuclear and nucleolar parameters as a database for diagnosis. The selected cases consisted of 16 primary and 12 metastatic neoplasms. It was determined that the P-Thy group had a significantly smaller nucleolar area (NuA) and nucleolar perimeter (NuP), and smaller SDs of nuclear area (NA), NuA, and NuP compared with the P-Ov and P-Other groups (P less than .05). The P-Ov group had a significantly smaller SD of NA compared with the P-Other group (P less than .05). The P-Ov group exhibited the greatest variability among the papillary neoplasms. Linear regression analysis indicated that in the P-Thy group alone there was a significant correlation between mean nuclear form factor (4 pi A/P2) and mean NuA (r = -.82; P less than .01), and mean NP and mean NuA (r = +.77; P less than .01). Linear regression analysis also indicated that in the P-Ov group alone, there was a significant correlation between mean NA and mean NuA (r = +.75; P less than .02). Morphometric domains were established using statistically significant sets of variables that distinguished between the groups. The application of three-dimensional computerized cluster analysis techniques indicated that the P-Thy group consistently had the smallest morphometric domains. It was concluded that ultrastructural morphometric analysis of papillary neoplasms has diagnostic potential and reveals interesting biological relationships among distinct nuclear features in the different groups of neoplasms.

Adenocarcinoma, Papillary↗