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J Thompson

Publications and source records attributed to J Thompson.

At least 325 records · Page 18Linked to original sources
PubMed1995

2 A crystal structure of an extracellular fragment of human CD40 ligand.

BACKGROUND: The CD40 ligand (CD40L) is a member of the tumor necrosis factor (TNF) family of proteins and is transiently expressed on the surface of activated T cells. The binding of CD40L to CD40, which is expressed on the surface of B cells, provides a critical and unique pathway of cellular activation resulting in antibody isotype switching, regulation of apoptosis, and B cell proliferation and differentiation. Naturally occurring mutations of CD40L result in the clinical hyper-IgM syndrome, characterized by an inability to produce immunoglobulins of the IgG, IgA and IgE isotypes. RESULTS: We have determined the crystal structure of a soluble extracellular fragment of human CD40L to 2 A resolution and with an R factor of 21.8%. Although the molecule forms a trimer similar to that found for other members of the TNF family, such as TNF alpha and lymphotoxin-alpha, and exhibits a similar overall fold, there are considerable differences in several loops including those predicted to be involved in CD40 binding. CONCLUSIONS: The structure suggests that most of the hyper-IgM syndrome mutations affect the folding and stability of the molecule rather than the CD40-binding site directly. Despite the fact that the hyper-IgM syndrome mutations are dispersed in the primary sequence, a large fraction of them are clustered in space in the vicinity of a surface loop, close to the predicted CD40-binding site.

Amino Acid Sequence↗
PubMed1995

Cloning of a new cation ATPase from Plasmodium falciparum: conservation of critical amino acids involved in calcium binding in mammalian organellar Ca(2+)-ATPases.

In order to study molecules that may be involved in pH gradient formation in Plasmodium, we have identified a novel cation-translocating ATPase (P-type ATPase) gene from P. falciparum (Pf). We report the full-length nucleotide and deduced amino acid (aa) sequences of this gene that we called PfATPase4. The PfATPase4 protein shares features with the different members of eukaryotic P-type ATPases, such as a similar transmembrane (TM) organization and aa identity in functionally important regions. Interestingly, the PfATPase4 protein possesses conserved aa involved in calcium binding in mammalian organellar Ca(2+)-ATPases.

Adenosine Triphosphatases↗
PubMed1995

Cloning, expression, sequence analysis, and site-directed mutagenesis of the Tn5306-encoded N5-(carboxyethyl)ornithine synthase from Lactococcus lactis K1.

The gene (ceo) encoding N5-(carboxyethyl)ornithine synthase (EC 1.5.1.24) has been isolated from the sucrose-nisin transposon Tn5306 of Lactococcus lactis K1, sequenced, and expressed at high level in Escherichia coli. The cloned enzyme has allowed the synthesis of the novel N omega-carboxypropyl amino acids N5-(1-carboxypropyl)-L-ornithine and N6-(1-carboxypropyl)-L-lysine. Comparison of the deduced amino acid sequence of N5-(1-carboxyethyl)-L-ornithine synthase (M(r) = 35,323) to the functionally analogous octopine and nopaline synthases from crown gall tumors showed surprisingly little similarity. However, N5-(1-carboxyethyl)-L-ornithine synthase and yeast saccharopine dehydrogenase exhibit homology at their N and C termini, which suggests that these two proteins constitute a distinct branch of the amino acid dehydrogenase superfamily. A centrally located 9-amino acid segment (GSGNVAQGA) in N5-(1-carboxyethyl)-L-ornithine synthase is virtually identical with a sequence present in the beta alpha beta-fold of the nucleotide binding domain of several microbial NADPH-dependent glutamate dehydrogenases. A much longer sequence of approximately 80 residues has significant similarity to alanine dehydrogenase. Substitution of arginine 15 of N5-(1-carboxyethyl)-L-ornithine synthase by lysine resulted in loss of enzyme activity.

Alanine Dehydrogenase↗
PubMed1995

The cea10 gene encodes a secreted member of the murine carcinoembryonic antigen family and is expressed in the placenta, gastrointestinal tract and bone marrow.

Although members of the carcinoembryonic antigen (CEA) family have been shown to convey cell adhesion in vitro, their location in some tissues contradicts such a function. As a basis for investigating their in vivo functions, we are characterizing the mouse CEA family. This paper describes the structure and expression of a new murine family member, cea10. Two full-length cDNA clones were isolated from a mouse colon library, whose deduced protein sequence comprises two immunoglobulin variable-like N-domains, directly followed by a short C-terminal domain indicating that this molecule is secreted. Although this domain organization suggests a closer relationship to the murine pregnancy-specific glycoproteins (PSG), which form a subgroup within the CEA family, sequence comparisons place Cea10 within the CEA subgroup. Overlapping cosmid clones containing the complete cea10 locus were mapped and the exons determined. No A2-like exon, characteristic for all other members of the murine CEA family, could be found. Sequences of the promoter and the first exon showed remarkably high similarity to the corresponding regions of bgp1 and bgp2, two other members of the murine CEA subgroup. Consensus sequences for two transcription factors (USF and an AP-2-like factor) that bind to the human BGP gene promoter were also present in the cea10 promoter and possibly convey expression of these genes in epithelial cells. RNase protection assays revealed transcriptional activity of cea10 in the colon and early placenta (10.5-12.5-day embryos) and to a lower extent in the small intestine, cecum, stomach, salivary glands and bone marrow. As some other CEA family members are deregulated in tumors, we quantified the expression levels of Cea10 transcripts in colonic mucosa and in adenomatous polyps of Min/+ mice. No differences in the steady-state levels of Cea10 mRNA could be found, suggesting that the Cea10 protein does not play a role in early tumor development. Taken together, Cea10 combines characteristic features of both CEA and PSG subgroup members in its structure and expression pattern.

Amino Acid Sequence↗
PubMed1995

beta-Cystathionase from Bordetella avium. Role(s) of lysine 214 and cysteine residues in activity and cytotoxicity.

beta-Cystathionase (EC 4.4.1.8) from Bordetella avium is a pyridoxal 5'-phosphate (PLP)-dependent enzyme that catalyzes the hydrolysis of L-cystine to yield pyruvic acid, NH3, and thiocysteine. The latter compound is highly toxic toward MC3T3-E1 osteogenic cells, rat osteosarcoma cells, and other cell lines maintained in tissue culture (Gentry-Weeks, C. R., Keith, J. M., and Thompson, J. (1993) J. Biol. Chem. 268, 7298-7314). Site-directed mutagenesis has established that lysine 214 of the sequence TKYVGGHSD, is primarily responsible for internal aldimine binding of PLP in the holoenzyme. Translation of the DNA sequence of the beta-cystathionase gene (metC) from B. avium, reveals 4 cysteine residues/enzyme subunit (M(r) = 42,600), and spectrophotometric analysis with 4,4'-dithiodipyridine showed that there were no disulfide linkages in the native protein. beta-Cystathionase is inhibited by sulfhydryl-reactive agents, including N-ethylmaleimide (NEM). To elucidate the mechanism of NEM inhibition, each of the 4 cysteine residues at positions 88, 117, 279, and 309 was individually replaced by alanine or glycine. The mutant proteins C88A, C117G, C279G, and C309A were purified to homogeneity, and each was assayed for enzyme activity, PLP-binding, NEM sensitivity, and susceptibility to chymotrypsin digestion. The activities of mutant proteins C88A and C279G were comparable with that of the native enzyme, and since both forms were inhibited by NEM, neither cysteine 88 nor 279 are prerequisite for enzyme activity. By elimination, cysteine residues 117 and 309 must be the targets for alkylation, and resultant inactivation of beta-cystathionase, by the -SH reactive agent. Substitution of cysteine 117 and 309 with glycine and alanine, respectively, yielded the inactive proteins C117G and C309A. PLP was not detectable in these proteins, and their absorption spectra lacked the peak (at 420 nm) that is characteristic of internal PLP-Schiff base formation. Edman degradation revealed that C117G (M(r) approximately 36,000) also lacked the first 63 amino acids comprising the N terminus of the native protein. The beta-cystathionase mutants C117G and C309A showed enhanced susceptibility to chymotrypsin digestion. Cysteine residues 117 and 309 may reside in conformationally sensitive environments, and in the native enzyme these amino acids most probably serve a structural function. Toxicity assays performed with the various mutant proteins obtained by site-directed mutagenesis established that only catalytically active forms of beta-cystathionase were were cytotoxic for tissue culture cells.

3T3 Cells↗
PubMed1995

Lymphocyte predominant Hodgkin's disease: a clinicopathologic comparative study of histologic and immunophenotypic subtypes.

PURPOSE: To compare the clinicopathologic features of the histologic and immunophenotypic subgroups of lymphocyte predominant Hodgkin's disease. METHODS AND MATERIALS: A retrospective review of 64 patients with lymphocyte predominant Hodgkin's disease treated at the Peter MacCallum Cancer Institute, Melbourne, was performed. Nodular and diffuse histological subtypes were confirmed by review of hematoxylin and eosin paraffin sections. Immunophenotyping with monoclonal antibodies L26 (B-cell origin) and Leu M1 (Hodgkin's phenotype) were available in 36 patients. RESULTS: The estimated freedom from progression and estimated overall survival at 10 years was 74% standard error (SE 5.8%) and 85% (SE 5.2%), non-Hodgkin's respectively. There were no significant differences in freedom from progression or overall survival when nodular and diffuse histology were compared. Similarly the presence of B-cell markers did not influence prognosis. There was only one case of secondary non-Hodgkin's lymphoma. CONCLUSION: Our results are consistent with major reported series displaying no differences between any of the subgroups of lymphocyte predominant Hodgkin's disease.

Adolescent↗
PubMed1995

Possible concentration-dependent suppression of immune response by verapamil.

Calcium channel blockers inhibit in vitro the lymphocyte response to mitogens and the generation of cytotoxic T-cell and natural killer cell activity. We report on a patient taking verapamil hydrochloride who experienced repeated, prolonged viral infections. His lymphocyte response to phytohemagglutinin, pokeweed mitogen, and tetanus antigen were suppressed; the mononuclear response to soluble tetanus was markedly diminished; and there was no response to influenza vaccination. On lowering the verapamil dose, or changing therapy to clonidine hydrochloride, all the above-mentioned functions returned to normal and a response to influenza vaccination was observed. We speculate that the prolonged viral infection observed in this patient may have been caused by suppression of the immune response by verapamil and we encourage further boad-based studies.

Aged↗
PubMed1995

Human intraperitoneal fibrinolytic response to elective surgery.

Intra-abdominal adhesions develop in over 90 per cent of patients undergoing laparotomy. Peritoneal fibrinolysis is believed to be important in the pathophysiology of adhesion formation. This study investigated the fibrinolytic response of postoperative peritoneal fluid in 12 patients undergoing elective laparotomy. There was a significant reduction in the plasminogen activating activity to undetectable levels at 24 h, which was sustained at 48 h (P < 0.05). While there was an early reduction in the concentration of tissue plasminogen activator (median 40.0, 28.2, 16.3 and 31.9 ng/ml at 2, 6, 24 and 48 h respectively; P < 0.05), the abolition of functional fibrinolytic activity appeared to be secondary to a marked increase in the concentration of plasminogen activator inhibitor (PAI) 1 (median 86, 196, 800 and 730 ng/ml at 2, 6, 24 and 48 h respectively; P < 0.05) and PAI-2 (median less than 6, 12, 155 and 245 ng/ml at 2, 6, 24 and 48 h respectively; P < 0.05). This reduction in the plasminogen activating activity of peritoneal fluid may favour the formation of permanent fibrous adhesions following surgery.

Adult↗
PubMed1995

Phorbol ester-induced priming of superoxide generation by phosphatidic acid-stimulated neutrophils and granule-free neutrophil cytoplasts.

This study was undertaken to examine the mechanisms involved in polymorphonuclear leukocyte superoxide release stimulated by exogenous phosphatidic acid (PA). Unlike the immediate burst of superoxide release affected by membrane-permeable dioctanoylglycerol (DiC8-DAG), dioctanoyl phosphatidic acid (DiC8-PA) induced superoxide release after a lag period of 5-20 min. This period was considerably reduced or eliminated when cells were primed by substimulatory levels of phorbol myristate acetate (PMA). Granule-depleted neutrophil cytoplasts also responded to DiC8-PA with a burst of superoxide generation. Activation of the cytoplast superoxide generating system in response to DiC8-PA was also significantly faster after cells had been preexposed to substimulatory levels of PMA, indicating that at least a portion of the priming mechanism was independent of PMA-induced degranulation. To further examine the potential mechanism of PMA priming of responses to PA, we evaluated the activity of neutrophil ecto-phosphatidic acid phosphohydrolase (ecto-PA phosphohydrolase), which generates diacylglycerol from exogenous PA. PMA priming had no discernable effect on the activity of this enzyme. In addition, propranolol, an inhibitor of PA phosphohydrolase, did not selectively inhibit PMA priming of neutrophil responses to DiC8-PA, indicating that priming did not result from acceleration of DiC8-PA hydrolysis. We therefore investigated the possibility that activation of protein kinase C was the basis of the primed response. Several semiselective protein kinase C inhibitors (calphostin C, H-7, and acylmethylglycerol) inhibited DiC8-DAG- and DiC8-PA-induced superoxide release as well as PMA-primed responses to approximately the same extent. These results are consistent with the hypothesis that neutrophil responses to phosphatidate are mediated by diglyceride generated by the action of ecto-PA phosphohydrolase. PMA priming does not result from increased catalytic activity of ecto-PA phosphohydrolase but rather seems to result from potentiation of an intermediate involved in the cells' response to multiple stimuli.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1995

Post-exercise gastric emptying of carbohydrate solutions determined using the 13C acetate breath test.

In an attempt to measure gastric emptying of carbohydrate solutions after exercise, we used the 13C acetate breath test to differentiate the gastric emptying of three approximately isoenergetic carbohydrate solutions (i.e. glucose, glucose polymer and sucrose) from each other and from water. On four separate occasions, six post-absorptive subjects walked on an inclined treadmill at 70% maximum oxygen uptake for 1 h and were then given 330 ml of one of the solutions in which 150 mg of sodium 1-[13C] acetate had been dissolved. Breath samples were collected at regular (2-30 min) intervals over the next 3.5 h for analysis of expired 13CO2 by isotope ratio mass spectrometry. When water was given, all subjects reached peak breath enrichment after 30 min, and had a mean (SE) gastric emptying time of 33.2 (1.6) min. Peak breath enrichment occurred later for sucrose and glucose polymer at 54.3 (3.1) min and 59.0 (2.1) min respectively (P < 0.01), and for glucose this was even later, at 62.3 (1.0) min (P < 0.05). Calculated gastric emptying times for sucrose and glucose polymer were almost identical [66.5 (2.5) and 69.8 (2.9) min respectively], whereas that for glucose was significantly slower [76.8 (3.2) min; P < 0.02], probably reflecting the effects of increased osmolality. The gastric emptying of all carbohydrates were significantly longer than for water (P < 0.01). These results show that in the post-exercise state the 13C acetate breath test can be used to differentiate the gastric emptying rates of water and carbohydrate solutions of different properties.

Acetates↗
PubMed1995

A human ubiquitin conjugating enzyme, L-UBC, maps in the Alzheimer's disease locus on chromosome 14q24.3.

We have identified a novel ubiquitin conjugating enzyme gene, L-UBC, which maps to human Chromosome (Chr) 14q24.3. This is also the location of the major early onset familial Alzheimer's disease gene (FAD3). L-UBC encodes a protein that demonstrates homology to the yeast ubiquitin conjugating enzyme, UBC-4, and human UbcH5. Their functions are to ubiquitinate specific proteins targeted for degradation. The protein also exhibits very strong homology to a rabbit protein, E2-F1, which mediates p53 degradation driven by papilloma virus E6 protein in vitro. The accumulation of specific proteins that have undergone aberrant processing in neurofibrillary tangles and amyloid plaques is the classic pathological feature in brains of Alzheimer's disease patients. Abnormal ubiquitination has previously been suggested to play a role in the etiology of Alzheimer's disease. This gene therefore represents a plausible candidate gene for FAD3.

Alzheimer Disease↗
PubMed1995

The Kings Cross fire: psychological reactions.

The psychological reactions of 50 survivors of the King's Cross fire, which hit London's underground railway system on the 18th day of November 1987, are described. Results are presented for the Impact of Event scale, the General Health Questionnaire (28-item version), the Eysenck Personality Questionnaire and a King's Cross Event Schedule. These are investigated in relation to (a) validity measures, (b) relationships between exposure and personality, and (c) spontaneous de-briefing. Two thirds of the sample had significant levels of psychological distress (meeting the "caseness" criterion on the GHQ). Both personality (neuroticism and L-scale) and degree of trauma exposure were related to subsequent reaction. Spontaneous debriefing was associated with subjective benefit. Transportation disasters present particular problems in relation to research and service delivery.

Adaptation, Psychological↗
PubMed1995

Sexual-stage-specific RNA expression of a new Plasmodium falciparum gene detected by in situ hybridisation.

A gene, Pf77, transcribed in the sexual stages of Plasmodium falciparum was isolated from a genomic expression library with a polyclonal antibody raised to gametocyte proteins. The entire coding region was obtained from a series of overlapping genomic and cDNA clones (from gametocyte RNA). A single open reading frame is present with no introns and no tandem repeat sequences. A Pf77 probe hybridised to a single transcript present in RNA prepared from purified gametocytes and could not be detected in RNA prepared from asexual blood stages. In situ hybridisation studies confirmed that the expression of Pf77 mRNA is sexual-stage-specific and in addition, showed that Pf77 mRNA is present only in female gametocytes during the vertebrate stages of the parasite's development.

Amino Acid Sequence↗