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Biomedical subjects

R J Harris

Publications and source records attributed to R J Harris.

At least 19 recordsLinked to original sources
PubMed2006

Predicting response to recombinant factor VIIa in non-haemophiliac patients with severe haemorrhage.

BACKGROUND: Despite increasing use of recombinant factor VIIa (rFVIIa) in non-haemophiliac patients, it is unclear when rFVIIa might be effective. METHODS: A single centre review of consecutive non-haemophiliac patients receiving rFVIIa for the management of severe haemorrhage. Treatments with rFVIIa were at a dose of 90 mug kg(-1) repeated at three hourly intervals at the clinicians' discretion. RESULTS: Eighteen patients received rFVIIa. Six patients survived to discharge and 12 patients died in hospital. The median (range) Sequential Organ Failure Assessment (SOFA) score at the time of administration of rFVIIa for the group that survived was 8.0 (5-12) compared with the group that died 12.0 (7.0-14.0) (P=0.03). One of the patients who survived (17%) had organ failure at the time of rFVIIa administration compared with 11 of those who died (92%) (P=0.004). Fifteen patients survived long enough to consider a second dose of rFVIIa, one patient who survived to discharge needed more than one dose (1/6, 17%), compared with seven of those who later died in hospital (7/9, 78%) (P=0.04). The survivors had a significant reduction in blood product requirements after rFVIIa, while patients who died did not. Neither the prothrombin time nor the activated partial thromboplastin time before or after rFVIIa predicted survival. CONCLUSIONS: High SOFA score and failure to respond to one adequate dose of rFVIIa appear to identify patients with poor prognosis. These observations may help in determining when rFVIIa treatment is likely to be futile.

Adolescent↗
PubMed2005

Zero-range model of traffic flow.

A multicluster model of traffic flow is studied, in which the motion of cars is described by a stochastic master equation. Assuming that the escape rate from a cluster depends only on the cluster size, the dynamics of the model is directly mapped to the mathematically well-studied zero-range process. Knowledge of the asymptotic behavior of the transition rates for large clusters allows us to apply an established criterion for phase separation in one-dimensional driven systems. The distribution over cluster sizes in our zero-range model is given by a one-step master equation in one dimension. It provides an approximate mean-field dynamics, which, however, leads to the exact stationary state. Based on this equation, we have calculated the critical density at which phase separation takes place. We have shown that within a certain range of densities above the critical value a metastable homogeneous state exists before coarsening sets in. Within this approach we have estimated the critical cluster size and the mean nucleation time for a condensate in a large system. The metastablity in the zero-range process is reflected in a metastable branch of the fundamental flux-density diagram of traffic flow. Our work thus provides a possible analytical description of traffic jam formation as well as important insight into condensation in the zero-range process.

Journal Article↗
PubMed2005

Autocrine VEGF mediates the antiapoptotic effect of CD154 on CLL cells.

CD154 is an important regulator of chronic lymphocytic leukaemia (CLL)-cell survival. In CLL, high serum levels of VEGF are a feature of advanced disease, and we and others have previously shown that CLL cells produce and secrete this growth factor. Since CD154 stimulates VEGF production in other cell types, and VEGF is known to promote cell survival, we examined whether the cytoprotection of CLL cells by CD154 involves VEGF. We report for the first time that treatment of CLL cells with CD154 results in increased VEGF production and demonstrate involvement of NF-kappaB in this process. Moreover, we show that CD154-induced CLL-cell survival is reduced by anti-VEGF-neutralising antibody and by inhibiting VEGF receptor (VEGFR) signalling with SU5416. However, addition of exogenous VEGF alone or blocking secreted autocrine VEGF had little or no effect on CLL-cell survival. We therefore conclude that CLL-cell cytoprotection in the presence of CD154 requires combined signalling by both CD40 and VEGFR. This combined signalling and resulting cytoprotection were shown to involve NF-kappaB activation and increased survivin production. In conclusion, our findings identify autocrine VEGF as an important mediator of the antiapoptotic effect of CD40 ligation, and thus provide new insights into CLL-cell rescue by CD154 in lymphoreticular tissues.

Apoptosis↗
PubMed2005

Long-term persistence of Coxiella burnetii after acute primary Q fever.

BACKGROUND: Long-term persistence of C. burnetii in infected animals was established in the 1950s and 60s, but the implications for human Q fever are not fully explored. AIM: To compare the prevalence of markers of infection in a cohort of Q fever patients in Australia (up to 5 years after infection) with those in the 1989 Birmingham cohort (12 years after infection). DESIGN: Case follow-up study. METHODS: C. burnetii was tested for by: (i) antibodies to Phase 1 and 2 antigens in the three immunoglobulin classes; (ii) detection of DNA in bone marrow and peripheral blood mononuclear cells by PCR assays directed against several different targets in the genome; and (iii) attempts to isolate coxiellas in cell culture or mice from PCR-positive samples. Amplicon specificity was verified by fluorometric probing and by sequencing. Cross-contamination was excluded by extensive use of non-template controls, and in particular by the use of certain IS1111a target sequences. RESULTS: Irrespective of clinical state, both groups remained seropositive, principally exhibiting medium levels of IgG antibody against C. burnetii Phase 2 antigen. C. burnetii genomic DNA was detected by PCR in 65% of bone marrow aspirates from Australian patients and approximately 88% of Birmingham patients. No coxiella were isolated from PCR positive samples. DISCUSSION: We propose a provisional model for persistence. In Q fever without sequelae, the process is largely confined to the bone marrow. In Q fever fatigue syndrome (QFS), it is modulated by the patient's immunogenetic background to give higher levels of coxiella genomes in bone marrow and increased shedding into the peripheral blood. In Q fever endocarditis, late pregnancy, or during iatrogenic or other immunosuppression, the multiplication cycle is prolonged, and a potential source of live organisms.

Animals↗
PubMed2005

Heterogeneity of recombinant antibodies: linking structure to function.

Structural heterogeneity of recombinant IgG1 antibodies derives from variations in conserved as well as unique structural features. Common sources of heterogeneity include Fc glycosylation, partial heavy chain C-terminal Lys processing, Fc methionine oxidation, hinge-region cleavage, and the glycation of Lys residues. Aspartate residues that are isomerized to iso-aspartate were detected by cation exchange or hydrophobic interaction chromatography for trastuzumab and omalizumab, respectively. Unpaired cysteines were detected in omalizumab using Ellman's reagent, with the thiol-containing Fab resolved using hydrophobic interaction chromatography after papain digestion. Structural variations that cause chromatographic resolution may indicate the presence of a form with reduced potency.

Animals↗
PubMed2004

Disordered asymmetric simple exclusion process: mean-field treatment.

We provide two complementary approaches to the treatment of disorder in a fundamental nonequilibrium model, the asymmetric simple exclusion process. First, a mean-field steady-state mapping is generalized to the disordered case, where it provides a mapping of probability distributions and demonstrates how disorder results in a new flat regime in the steady-state current-density plot for periodic boundary conditions. This effect was earlier observed by Phys. Rev. E 58, 1911 (1998)] but we provide a treatment for more general distributions of disorder, including both numerical results and analytic expressions for the width 2 Delta(C) of the flat section. We then apply an argument based on moving shock fronts [Europhys. Lett. 48, 257 (1999)]] to show how this leads to an increase in the high-current region of the phase diagram for open boundary conditions. Second, we show how equivalent results can be obtained easily by taking the continuum limit of the problem and then using a disordered version of the well-known Cole-Hopf mapping to linearize the equation. Within this approach we show that adding disorder induces a localization transformation (verified by numerical scaling), and Delta(C) maps to an inverse localization length, helping to give a physical interpretation to the problem.

Journal Article↗
PubMed2003

Variation in immune response genes and chronic Q fever. Concepts: preliminary test with post-Q fever fatigue syndrome.

Acute primary Q fever is followed by various chronic sequelae. These include subacute Q fever endocarditis, granulomatous reactions in various organs or a prolonged debilitating post-infection fatigue syndrome (QFS). The causative organism, Coxiella burnetii, persists after an initial infection. The differing chronic outcomes may reflect variations within cytokine and accessory immune control genes which affect regulation of the level of persistence. As a preliminary test of the concept we have genotyped QFS patients and controls for gene variants spanning 15 genes and also examined HLA-B and DR frequencies. QFS patients exhibited a significantly increased frequency of HLA-DR-11 compared with controls and also significant differences in allelic variant frequencies within the NRAMP, and IFNgamma genes. These results indicate a possible genetic role in the expression of overt chronic Q fever. Further studies will be undertaken to increase sample sizes, to survey other forms of chronic Q fever and to examine Q fever patients who have recovered without sequelae.

Cation Transport Proteins↗
PubMed2001

Modification of epidermal growth factor-like repeats with O-fucose. Molecular cloning and expression of a novel GDP-fucose protein O-fucosyltransferase.

The O-fucose modification is found on epidermal growth factor-like repeats of a number of cell surface and secreted proteins. O-Fucose glycans play important roles in ligand-induced receptor signaling. For example, elongation of O-fucose on Notch by the beta1,3-N-acetylglucosaminyltransferase Fringe modulates the ability of Notch to respond to its ligands. The enzyme that adds O-fucose to epidermal growth factor-like repeats, GDP-fucose protein O-fucosyltransferase (O-FucT-1), was purified previously from Chinese hamster ovary (CHO) cells. Here we report the isolation of a cDNA that encodes human O-FucT-1. A probe deduced from N-terminal sequence analysis of purified CHO O-FucT-1 was used to screen a human heart cDNA library and expressed sequence tag and genomic data bases. The cDNA contains an open reading frame encoding a protein of 388 amino acids with a predicted N-terminal transmembrane sequence typical of a type II membrane orientation. Likewise, the mouse homolog obtained from an expressed sequence tag and 5'-rapid amplification of cDNA ends of a mouse liver cDNA library encodes a type II transmembrane protein of 393 amino acids with 90.4% identity to human O-FucT-1. Homologs were also found in Drosophila and Caenorhabditis elegans with 41.2 and 29.4% identity to human O-FucT-1, respectively. The human gene (POFUT1) is on chromosome 20 between PLAGL2 and KIF3B, near the centromere at 20p11. The mouse gene (Pofut1) maps near Plagl2 on a homologous region of mouse chromosome 2. POFUT1 gene transcripts were expressed in all tissues examined, consistent with the widespread localization of the modification. Expression of a soluble form of human O-FucT-1 in insect cells yielded a protein of the predicted molecular weight with O-FucT-1 kinetic and enzymatic properties similar to those of O-FucT-1 purified from CHO cells. The identification of the gene encoding protein O-fucosyltransferase I now makes possible mutational strategies to examine the functions of the unusual O-fucose post-translational modification.

Amino Acid Sequence↗
PubMed2001

Sexually transmitted Q fever.

We report the sexual transmission of Coxiella burnetii from a man with occupationally acquired Q fever to his wife. Fifteen days after coitus, his wife also developed serologically proven acute Q fever. C. burnetii DNA sequences were detected by polymerase chain reaction (PCR) performed on semen samples obtained from the husband at 4 and 15 months after the onset of acute Q fever, but PCR results were variable at 23 months, indicating the presence of few organisms.

Antibodies, Bacterial↗
PubMed2001

Identification of multiple sources of charge heterogeneity in a recombinant antibody.

Seven forms of a therapeutic recombinant antibody that binds to the her2/neu gene product were resolved by cation-exchange chromatography. Structural differences were assigned by peptide mapping and HIC after papain digestion. Deamidation of light chain asparagine 30 to aspartate in one or both light chains is responsible for two acidic forms. A low potency form is due to isomerization of heavy chain aspartate 102; the Asp102 succinimide is also present in a basic peak fraction. Forms with both Asn30 deamidation and Asp102 isomerization modifications were isolated. Deamidation of heavy chain Asn55 to isoaspartate was also detected. Isoelectric focusing in a polyacrylamide gel was used to verify the assignments. All modifications were found in complementarity determining regions.

Amino Acid Sequence↗
PubMed2001

Speed of collapse of the non-ventilated lung during single-lung ventilation for thoracoscopic surgery: the effect of transient increases in pleural pressure on the venting of gas from the non-ventilated lung.

A study of 10 anaesthetised patients placed in the lateral position for thoracoscopic surgery assessed whether transient increases in pleural pressure on the side of the non-ventilated lung might increase the speed at which gas vents from that lung. The transient increases in pleural pressure were generated by the mediastinal displacement that occurs with each inspiratory phase of positive pressure ventilation of the dependent lung. When combined with a unidirectional valve allowing gas to flow out of the non-ventilated lung, and a second valve allowing ambient airflow into, but not out of, the thoracic cavity via an initial thoracoscopy access site, this mediastinal displacement could conceivably serve to 'pump' gas out of the non-ventilated lung. Using the four different combinations of valve inclusion or omission, the volume of gas that vented from the non-ventilated lung into a measuring spirometer was recorded during a 120-s measurement sequence. It was found that the speed of venting was not increased by the transient increases in pleural pressure, and that in all but one of a total of 34 measurement sequences, venting had ceased by the end of the sequence. Gas venting was a mean (SD) of 85.5 (11.9)% complete in 25 s (five breaths), and 96.6 (6.1)% complete in 60 s. This prompt partial lung collapse very likely reflected the passive elastic recoil of the lung, while the failure of transient increases in pleural pressure to result in ongoing venting of gas was probably a consequence of airways closure as the lung collapsed. It is concluded that techniques that aim to speed lung collapse by increasing pleural pressure are unlikely to be effective.

Adolescent↗
PubMed2001

The inhibition of platelet aggregation and blood coagulation by Micropechis ikaheka venom.

Uncoagulable blood and life-threatening bleeding can result from the action of some snake venom toxins on haemostatic components of blood and vessel walls. Although envenoming by Micropechis ikaheka primarily affects neurones and muscle cells causing post-synaptic neuromuscular blockade and rhabdomyolysis, disturbances of haemostasis also occur. Therefore, the present study explored the effects of M. ikaheka venom on platelets and endothelium, which are important components of the haemostatic mechanism. The venom inhibited platelet aggregation in response to ADP and collagen, and also delayed clotting dependent on platelet activation or endothelial cell tissue factor expression. Some of these effects were reduced by the incubation of venom with a phospholipase A2 (PLA2) inhibitor and could be reproduced by a 17 kDa venom fraction containing a PLA2. In addition, an 11 kDa fraction containing a long-chain neurotoxin reduced ADP-induced aggregation. The venom was also found to reduce endothelial cell adherence to vitronectin-, fibronectin- and collagen-coated surfaces. These results suggest that, by inhibiting procoagulant activities of platelets and endothelial cells, a 17 kDa PLA2 plays an important role in the anticoagulant action of M. ikaheka venom.

Anticoagulants↗
PubMed2001

Organization of the genes involved in dimethylglycine and sarcosine degradation in Arthrobacter spp.: implications for glycine betaine catabolism.

The nucleotide sequences of two cloned DNA fragments containing the structural genes of heterotetrameric sarcosine oxidase (soxBDAG) and dimethylglycine dehydrogenase (dmg) from Arthrobater spp. 1-IN and Arthrobacter globiformis, respectively, have been determined. Open reading frames were identified in the soxBDAG operon corresponding to the four subunits of heterotetrameric sarcosine oxidase by comparison with the N-terminal amino-acid sequences and the subunit relative molecular masses of the purified enzyme. Alignment of the deduced sarcosine oxidase amino-acid sequence with amino-acid sequences of functionally related proteins indicated that the arthrobacterial enzyme is highly homologous to sarcosine oxidase from Corynebacterium P-1. Deletion and expression analysis, and alignment of the deduced amino-acid sequence of the dmg gene, showed that dmg encodes a novel dimethylglycine oxidase, which is related to eukaryotic dimethylglycine dehydrogenase, and contains nucleotide-binding, flavinylation and folate-binding motifs. The recombinant dimethylglycine oxidase was purified to homogeneity and characterized. The DNA located upstream and downstream of both the soxBDAG and dmg genes is predicted to encode enzymes involved in the tetrahydrofolate-dependent assimilation of methyl groups. Based on the sequence analysis reported herein, pathways are proposed for glycine betaine catabolism in Arthrobacter species, which involve the identified folate-dependent enzymes.

Amino Acid Sequence↗
PubMed2001

Clinical evaluation of 3 techniques to augment keratinized tissue without root coverage.

BACKGROUND: The importance of keratinized tissue is a controversial subject. However, in some situations most clinicians would agree that surgical procedures to increase the amount of keratinized tissue without root coverage are indicated. In this study, 3 surgical procedures were compared in their ability to increase the width of keratinized tissue. They are: the epithelized autogenous masticatory mucosa graft (free gingival graft), autogenous predominately connective tissue graft (connective tissue graft), and acellular dermal matrix. METHODS: Forty-five patients referred for treatment of areas with inadequate keratinized tissue were randomly assigned into 1 of 3 groups of 15 each. Each group was treated with 1 of the 3 surgical procedures to increase the width of keratinized tissue. The width of keratinized tissue pre- and postsurgery was evaluated. RESULTS: All 3 groups started with a similar width of keratinized tissue. All of the surgical procedures resulted in a statistically significant increase in the width of keratinized tissue: free gingival graft, 4.1 mm; connective tissue graft, 3.6 mm; and acellular dermal matrix, 4.1 mm. CONCLUSION: A statistically significant increase in the amount of keratinized tissue was obtained with all 3 surgical procedures evaluated.

Adolescent↗
PubMed2001

Root coverage of a palatal recession defect: a case report.

BACKGROUND: Obtaining predictable root coverage has become an important part of periodontal therapy. However, little has been reported concerning the treatment of palatal recession defects. This is a case report which demonstrates a technique to obtain root coverage of a palatal recession defect. METHODS: The technique involved elevating a partial-thickness palatal flap and placing a connective tissue graft beneath the flap and over the recession defect (subepithelial graft). The portion of the connective tissue graft placed over the recession defect was not covered by the flap. RESULTS: Clinically significant amounts of root coverage were obtained (84.6%), although the connective tissue graft was not covered by the flap. CONCLUSION: Root coverage of a palatal recession defect can be obtained with a subepithelial graft.

Adult↗
PubMed2001

The effect of lexical, pragmatic, and morphological violations on reading time and deviance ratings of English and German sentences.

Effects on reading time and deviance ratings of word-choice violations were studied at the surface code, propositional textbase, and situation model levels of representation in German and English sentences. Lexical violations (propositional textbase level) such as The housewife massaged the bread dough were rated as more deviant from normal language than were pragmatic violations (situation model level) such as The police officer shot the parking violator, although the pragmatic violations took longer to comprehend. The addition of a morphological (surface code) violation through the wrong form of an article (e.g., a ugly car) decreased reading time but only in cases where that material was deeply processed and where the morphology carried substantial syntactic information (e.g., in German, the definite article carries case, gender, and number information). The results thus confirmed the operation of comprehension strategies at multiple levels of representation and identified some cross-linguistic generality in comprehension and some language differences.

Adolescent↗