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Biomedical subjects

A J Morris

Publications and source records attributed to A J Morris.

At least 127 records · Page 7Linked to original sources
PubMed1997

Characterization of two alternately spliced forms of phospholipase D1. Activation of the purified enzymes by phosphatidylinositol 4,5-bisphosphate, ADP-ribosylation factor, and Rho family monomeric GTP-binding proteins and protein kinase C-alpha.

We previously reported the cloning of a cDNA encoding human phosphatidylcholine-specific phospholipase D1 (PLD1), an ADP-ribosylation factor (ARF)-activated phosphatidylcholine-specific phospholipase D (Hammond, S. M., Tsung, S., Autschuller, Y., Rudge, S. A., Rose, K., Engebrecht, J., Morris, A. J., and Frohman, M. A. (1995) J. Biol. Chem. 270, 29640-29643). We have now identified an evolutionarily conserved shorter splice variant of PLD1 lacking 38 amino acids (residues 585-624) that arises from regulated splicing of an alternate exon. Both forms of PLD1 (PLD1a and 1b) have been expressed in Sf9 cells using baculovirus vectors and purified to homogeneity by detergent extraction and immunoaffinity chromatography. PLD1a and 1b have very similar properties. PLD1a and 1b activity is Mg2+dependent but insensitive to changes in free Ca2+ concentration. Phosphatidylinositol 4,5-bisphosphate and phosphatidylinositol 3,4,5-trisphosphate activate PLD1a and 1b but a range of other acidic phospholipids are ineffective. PLD1a and 1b are highly responsive to activation by GTP-gammaS-liganded ADP-ribosylation factor-1 (ARF-1) and can also be activated to a lesser extent by three purified RHO family monomeric GTP-binding proteins, RHO A, RAC-1, and CDC42. Activation of PLD1a and 1b by the RHO family monomeric GTP-binding proteins is GTP-dependent and synergistic with ARF-1. Purified protein kinase C-alpha activates PLD1a and 1b in a manner that is stimulated by phorbol esters and does not require ATP. Activation of PLD1a and 1b by protein kinase C-alpha is synergistic with ARF and with the RHO family monomeric GTP-binding proteins, suggesting that these three classes of regulators interact with different sites on the enzyme.

Adenosine Diphosphate Ribose↗
PubMed1997

Bioprocess supervision: neural networks and knowledge based systems.

Supervision of highly non-linear and time variant bioprocesses is of considerable importance to bioindustries as a means of achieving improved productivity and reduced process variability. Artificial intelligence methodologies, including artificial neural networks and knowledge based systems, can contribute significantly to the achievement of these objectives.

Biosensing Techniques↗
PubMed1997

Comparison of selection based on phenotype, selection index and best linear unbiased prediction using data from a closed broiler line.

1. Selection based on three methods of estimating breeding values, Best Linear Unbiased Prediction (BLUP), selection index (SI), and phenotype (SP) were compared for three traits, juvenile body weight (JW), percentage breast meat yield (BM) and hen-day rate of egg production (EP) using records provided by a commercial broiler breeding company. 2. Product moment correlations were calculated between breeding values estimated by each method and averaged across sexes. A mean correlation of 0.69 was obtained between selection on SP and BLUP for JW. Mean correlations of 0.88 and 0.68 and 0.87 were obtained between SI and BLUP for the traits JW, EP and BM, respectively. 3. A mean estimated genetic response of 77.7% was obtained with SP for JW relative to BLUP in the absence of restrictions on the selection of close relatives. Estimated genetic responses of 90.7%, 66.9% and 88.4% were obtained by SI relative to BLUP for JW, EP and BM, respectively. 4. Applying restrictions on the selection of close relatives resulted in slight decreases in estimated responses but not in the respective ranking of the selection methods. 5. The results indicate that BLUP could provide commercial breeders with increased selection responses compared to index selection, in particular for traits of low heritability and where relatively few animals possess performance records.

Animals↗
PubMed1997

Assessment of the yield of anaerobic blood cultures.

Because of the declining incidence of anaerobic bacteremia, the predictable sites of anaerobic infection and the increasing importance of aerobic isolates (eg; yeasts), the practice of routinely culturing half the volume of blood collected anaerobically has been questioned. We have assessed the yield of routine anaerobic blood cultures in our clinical setting. Blood culture isolates from November 1994 through October 1995 at Auckland (AKH) and Green Lane/National Women's Hospitals (GL/NWH) were recorded. The medical records of patients with anaerobic bacteremia were examined. For the three month period April to June 1996, all positive blood cultures were analysed with respect to which bottle (aerobic or anaerobic or both) was positive. For the period November 1994 to October 1995, 5.6% and 5.3% of blood cultures at AKH and GLH respectively were positive. At AKH and GLH anaerobes constituted 0.16% and 0.19% of all blood cultures and 3.1% and 3.5% of all positive blood cultures respectively. Twenty-one of 25 (84%) significant anaerobes were from patients in whom anaerobic infection was predictable. More isolates were recovered from aerobic than anaerobic bottles, 178 versus 71, p < 0.001. Aerobic culture also recovered more pathogens (76 versus 38, p < 0.001 more yeasts (10 versus 0) and more Pseudomonas spp. (10 versus 1) than did anaerobic culture. Only obligate anaerobes were isolated more frequently in anaerobic bottles (5 versus 0, p = 0.03). Most instances of anaerobic bacteremia occurred in patients where anaerobes could be expected. We conclude that routine use of two aerobic bottles with clinically directed use of anaerobic blood culture bottle is an appropriate and effective approach in our setting.

Bacteremia↗
PubMed1997

Bartonella henselae bacteraemia in domestic cats from Auckland.

Bartonella henselae causes most cases of cat scratch disease, a self-limited localised lymphadenopathy illness of humans. Bartonella henselae also causes disseminated cutaneous and visceral disease in immunocompromised people. Cat blood (1-5 ml) collected from cats in the Auckland area was processed and plated on to 5% sheep blood brain heart infusion agar and incubated at 35 degrees C in 5% CO2 for 14 days. Bartonella henselae was identified by colony morphology, Gram's stain, twitching motility, biochemical tests and molecular methods. Eight of 48 cats (17%) had Bartonella bacteraemia. Species-specific probes and biochemical profiles identified all isolates as B. henselae. Infected cats pose a risk to humans they lick, scratch or bite. People should be made aware of the risk cats pose.

Journal Article↗
PubMed1997

Candidal mediastinitis: an emerging clinical entity.

Candidal mediastinitis is rare. We report nine cases encountered at our institutions since 1985; seven cases were diagnosed since 1993. All cases followed thoracic surgery, with a median time from surgery to disease onset of 11 days (range, 6-100 days). All patients received prior antibiotic therapy. Common clinical manifestations were chest wall erythema in 4 cases (44%), drainage in 5 (56%), fever in 4 (44%), and sternal instability in 4 (44%). Failure to obtain appropriate intraoperative specimens for cultures and the dismissal of cultures positive for Candida as contaminants delayed diagnosis in three cases (33%). Mediastinitis was complicated by contiguous or hematogenous spread in seven cases (78%); five patients (56%) had two or more complications. The mortality rate was 56%. Optimal therapy remains undefined, but on the basis of our experience, aggressive surgical debridement combined with antifungal therapy for at least 6 weeks is recommended. Prompt recognition and institution of therapy appear to be the keys to improving prognosis.

Aged↗
PubMed1997

Transfusion transmitted Yersinia enterocolitica infection in New Zealand.

AIMS: To update and summarise cases of transfusion-transmitted Yersinia enterocolitica infection in New Zealand and to evaluate critically suggested methods to reduce this rare but frequently fatal complication of blood transfusion. METHODS: Case reports of four recent transfusion-transmitted Y. enterocolitica infections in New Zealand are given and previous reports reviewed. Literature review and evaluation of proposed methods to decrease the incidence of transfusing yersinia contaminated blood. RESULTS: There have been eight cases of transfusion-transmitted Y. enterocolitica infection in New Zealand in the past five years. Four of the five deaths have been directly caused by the transfusion. This gives a transfusion incidence rate of one:65,000 and a fatality rate of one:104,000 units transfused. This fatality rate is more than 80 times higher than that reported in the United States. CONCLUSIONS: Why the incidence of transfusion-transmitted yersinia is so high is not clear, since we do not store blood as long as many other countries, particularly the United States. In Auckland, however, the cases came at a time when the number of yersinia isolates from the community is reported to be rising. Many suggestions for the prevention of this problem have been put forward reflecting the fact that there is as yet no perfect solution. Those which are easy to implement and cheap to perform are largely already in place and investigation is continuing into the other alternatives.

Adolescent↗
PubMed1997

Comparison of PCR, culture, and histopathology for diagnosis of tuberculous pericarditis.

Nucleic acid amplification techniques for the diagnosis of tuberculosis (TB) are rapidly being developed. Scant work, however, has focused on pericardial TB. Using cryopreserved specimens from a prior study of pericarditis, we compared PCR to culture and histopathology for the diagnosis of tuberculous pericarditis in 36 specimens of pericardial fluid and 19 specimens of pericardial tissue from 20 patients. Fluid and tissue were cultured on Lowenstein-Jensen and Middlebrook solid media and in BACTEC radiometric broth. Tissue specimens were stained with hematoxylin-eosin, Ziehl-Neelsen, auramine O, and Kinyoun stains and were examined for granuloma formation and acid-fast bacilli. PCR was performed with both fluid and tissue with IS6110-based primers specific for the Mycobacterium tuberculosis complex by published methods. Sixteen of the 20 patients had tuberculous pericarditis and 4 patients had other diagnoses. TB was correctly diagnosed by culture in 15 (93%) patients, by PCR in 13 (81%) patients, and by histology in 13 of 15 (87%) patients. PCR gave one false-positive result for a patient with Staphylococcus aureus pericarditis. Considering the individual specimens as the unit of analysis, M. tuberculosis was identified by culture in 30 of 43 specimens (70%) from patients with tuberculous pericarditis and by PCR in 14 of 28 specimens (50%) from patients with tuberculous pericarditis (P > 0.15). The sensitivity of PCR was higher with tissue specimens (12 of 15; 80%) than with fluid specimens (2 of 13; 15%; P = 0.002). In conclusion, the overall accuracy of PCR approached the results of conventional methods, although PCR was much faster. Therefore, PCR merits further development in this regard. The sensitivity of PCR with pericardial fluid was poor, and false-positive results with PCR remain a concern.

Adolescent↗
PubMed1997

Nosocomial acquisition of Burkholderia gladioli in patients with cystic fibrosis.

Burkholderia gladioli has been reported as colonizing the airways of patients with cystic fibrosis (CF) but has not previously been associated with adverse outcome. We describe six patients with CF in whom the same strain of B. gladioli, on the basis of ribotyping and biochemical characteristics, was grown in their sputum. Acquisition of this organism was followed by a fatal outcome in all six patients; one had a rapid decline in respiratory status and another developed fulminant B. gladioli bacteremia. Evidence suggests that patient-to-patient transmission of the organism occurred, and supports nosocomial infection in the ward and/or outpatient clinic despite general and stringent infection-control measures. This is the first report of adverse clinical outcome following sputum colonization with B. gladioli, and the first to demonstrate person-to-person transmission.

Bacteremia↗
PubMed1997

The small guanosine triphosphate-binding protein Rab4 is involved in insulin-induced GLUT4 translocation and actin filament rearrangement in 3T3-L1 cells.

Insulin's stimulation of glucose transport involves the translocation of vesicles containing the glucose transporter GLUT4 to the plasma membrane. Small GTP-binding proteins have been implicated in the regulation of vesicular traffic. We studied the effects of microinjection of wild-type Rab4 glutathione S-transferase fusion protein (WT Rab4), a GTP-binding defective mutant (Rab4 N121I), a guanosine triphosphatase-defective mutant (Rab4 Q67L), and a Rab4 antibody on insulin-induced GLUT4 translocation in 3T3-L1 adipocytes. Microinjection of Rab4 N121I and Rab4 antibodies had no effect on basal GLUT4 staining, but inhibited insulin-induced GLUT4 translocation by 50% compared with that in control IgG-injected cells. WT Rab4 and Rab4 Q67L microinjection had no effect on either basal or insulin-induced GLUT4 translocation. Premixing and coinjection of the Rab4 antibody with WT Rab4 almost completely abolished its inhibitory effect on insulin-induced GLUT4 translocation. In contrast, microinjection of an antibody directed against the highly conserved region of Rab3 proteins had no effect on insulin-induced GLUT4. These results point to a direct role of Rab4 in insulin-induced GLUT4 translocation, and that this effect is dependent on nucleotide binding to the protein. We also studied the effect of microinjection of the same proteins on insulin-induced actin filament rearrangement (membrane ruffling) in the same cell line. Microinjection of Rab4 N121I and Rab4 antibodies inhibited insulin-induced membrane ruffling by 40%, whereas WT Rab4 or a Rab3 antibody injection had no effect on cytoskeletal rearrangement. In summary, 1) Rab4 is a necessary component of the insulin/GLUT4 translocation signaling pathway; 2) the function of Rab4 in this pathway requires GTP binding; 3) Rab4 also participates in the process of insulin-induced membrane ruffling; and 4) Rab3 proteins do not seem to be involved in these processes.

3T3 Cells↗
PubMed1996

Evidence for an insulin receptor substrate 1 independent insulin signaling pathway that mediates insulin-responsive glucose transporter (GLUT4) translocation.

Interaction of the activated insulin receptor (IR) with its substrate, insulin receptor substrate 1 (IRS-1), via the phosphotyrosine binding domain of IRS-1 and the NPXY motif centered at phosphotyrosine 960 of the IR, is important for IRS-1 phosphorylation. We investigated the role of this interaction in the insulin signaling pathway that stimulates glucose transport. Utilizing microinjection of competitive inhibitory reagents in 3T3-L1 adipocytes, we have found that disruption of the IR/IRS-1 interaction has no effect upon translocation of the insulin-responsive glucose transporter (GLUT4). The activity of these reagents was demonstrated by their ability to block insulin stimulation of two distinct insulin bioeffects, membrane ruffling and mitogenesis, in 3T3-L1 adipocytes and insulin-responsive rat 1 fibroblasts. These data suggest that phosphorylated IRS-1 is not an essential component of the metabolic insulin signaling pathway that leads to GLUT4 translocation, yet it appears to be required for other insulin bioeffects.

3T3 Cells↗
PubMed1996

Rho is only ARF the story. Phospholipid signalling.

The GTP-binding proteins ARF and Rho control a number of important cellular processes, such as protein traffic and cell morphology; there is increasing evidence that phospholipase D is a key mediator of these ARF/Rho-regulated events.

ADP-Ribosylation Factors↗