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J Brearley

Publications and source records attributed to J Brearley.

7 recordsLinked to original sources

Overexpression of auxin-binding protein enhances the sensitivity of guard cells to auxin.

To explore the role of auxin-binding protein (ABP1) in planta, a number of transgenic tobacco (Nicotiana tabacum) lines were generated. The wild-type KDEL endoplasmic reticulum targeting signal was mutated to HDEL, another common retention sequence in plants, and to KEQL or KDELGL to compromise its activity. The auxin-binding kinetics of these forms of ABP1 were found to be similar to those of ABP1 purified from maize (Zea mays). To test for a physiological response mediated by auxin, intact guard cells of the transgenic plants were impaled with double-barreled microelectrodes, and auxin-dependent changes in K(+) currents were recorded under voltage clamp. Exogenous auxin affected inwardly and outwardly rectifying K(+) currents in a dose-dependent manner. Auxin sensitivity was markedly enhanced in all plants overexpressing ABP1, irrespective of the form present. Immunogold electron microscopy was used to investigate the localization of ABP1 in the transgenic plants. All forms were detected in the endoplasmic reticulum and the KEQL and KDELGL forms passed further across the Golgi stacks than KDEL and HDEL forms. However, neither electron microscopy nor silver-enhanced immunogold epipolarization microscopy revealed differences in cell surface ABP1 abundance for any of the plants, including control plants, which indicated that overexpression of ABP1 alone was sufficient to confer increased sensitivity to added auxin. Jones et al. ([1998] Science 282: 1114-1117) found increased cell expansion in transgenic plants overexpressing wild-type ABP1. Single cell recordings extend this observation, with the demonstration that the auxin sensitivity of guard cell K(+) currents is mediated, at least in part, by ABP1.

Blotting, Northern↗

Tuberculosis surveillance in immigrants through health undertakings in Western Australia.

SETTING: Tuberculosis Control Program, Western Australia. OBJECTIVE: To assess the effectiveness and efficiency of tuberculosis (TB) surveillance in immigrants. DESIGN: Retrospective descriptive analysis of records of immigrants who first registered with the Tuberculosis Program from January 1994 to December 1995, having entered Western Australia on health undertakings (nonlegal contracts signed by those determined during premigration screening to require post-arrival evaluation). RESULTS: Of 1,344 immigrants on health undertakings for TB in the period, 587 (44%) had findings directly related to TB through pre-migration screening, 69 of whom required treatment for active disease. Another 443 (33%) had chest X-ray changes for which TB could not be excluded. Of the remaining 314 (23%), 172 had poor quality X-rays. Post-arrival assessment and follow-up of the whole group detected four of seven additional cases of active TB, 373 persons (28%) requiring on-going surveillance, 667 (50%) with non-TB conditions and 280 (21%) with normal chest X-rays. CONCLUSIONS: Migrant surveillance for TB in Australia is effective in detecting active disease and identifying a high-risk subgroup requiring further evaluation. Efficiency can be improved and high compliance achieved with simple administrative changes. Routine pre-migration Mantoux testing is inappropriate, and its inclusion in post-arrival assessments should be a national policy decision.

Adolescent↗

K+ channels of Cf-9 transgenic tobacco guard cells as targets for Cladosporium fulvum Avr9 elicitor-dependent signal transduction.

The Cf-9 gene encodes an extracytosolic leucine-rich repeat (LRR) protein that is membrane anchored near its C-terminus. The protein confers resistance in tomato to races of the fungus Cladosporium fulvum expressing the corresponding avirulence gene Avr9. In Nicotiana tabacum the Cf-9 transgene confers sensitivity to the Avr9 elicitor, and leads on elicitation to a subset of defence responses qualitatively similar to those normally seen in the tomato host. One of the earliest responses, both in the native and transgenic hosts, results in K+ salt loss from the infected tissues. However, the mechanism(s) underlying this solute flux and its control is poorly understood. We have explored the actions of Avr9 on Cf-9 transgenic Nicotiana using guard cells as a model. Much detail of guard cell ion channels and their regulation is already known. Measurements were carried out on intact guard cells in epidermal peels, and the currents carried by inward- (IK,in) and outward-rectifying (IK,out) K+ channels were characterized under voltage clamp. Exposures to Avr9-containing extracts resulted in a 2.5- to 3-fold stimulation of IK,out and almost complete suppression of IK,in within 3-5 min. The K+ channel responses were irreversible. They were specific for the Avr9 elicitor, were not observed in guard cells of Nicotiana lacking the Cf-9 transgene and, from kinetic analyses, could be ascribed to changes in channel gating. Both K+ channel responses were found to be saturable functions of Avr9 concentration and were completely blocked in the presence of 0.5 microM staurosporine and 100 microM H7, both broad-range protein kinase antagonists. These results demonstrate the ability of the Cf-9 transgene to couple Avr9 elicitation specifically to a concerted action on two discrete K+ channels and they indicate a role for protein phosphorylation in Avr9/Cf-9 signal transduction leading to transport control.

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

Preventive therapy for tuberculosis in Western Australia.

SETTING: State Tuberculosis Control Programme, Western Australia. OBJECTIVE: To ascertain baseline information, applicability and efficacy of preventive therapy for tuberculosis (TB) under indirectly supervised treatment in Western Australia. DESIGN: Retrospective analyses of records of persons with TB infection who were prescribed preventive therapy for the period 1993-1996 inclusive, using simple descriptive statistics. RESULTS: Preventive therapy was given to 411 persons after screening for TB due to on-arrival migrant surveillance (269), contact tracing (59), tuberculin surveys (59) and other reasons (24). Six-month isoniazid monotherapy (HMT) was prescribed for 403 and multidrug regimens for the remaining eight. Excepting 34 whose compliance was unknown, varying degrees of treatment non-adherence were found in 90 (24%) of the remaining 369 persons given HMT, including 36 (10%) with under five months of total medication. Minor adverse drug effects occurred in 32 (9%) subjects and contributed to the non-adherence in 23 of these. One person has since developed active TB. CONCLUSIONS: Preventive therapy in Western Australia conformed to generally accepted guidelines. Varying degrees of non-adherence to HMT occurred in 24% of persons, but 90% completed adequate therapy under indirect supervision. Non-adherence is significantly related to adverse drug effects.

Adolescent↗

Sensitivity to abscisic acid of guard-cell K+ channels is suppressed by abi1-1, a mutant Arabidopsis gene encoding a putative protein phosphatase.

Abscisic acid (ABA) modulates the activities of three major classes of ion channels--inward- and outward-rectifying K+ channels (IK,in and IK,out, respectively) and anion channels--at the guard-cell plasma membrane to achieve a net efflux of osmotica and stomatal closure. Disruption of ABA sensitivity in wilty abi1-1 mutants of Arabidopsis and evidence that this gene encodes a protein phosphatase suggest that protein (de)-phosphorylation contributes to guard-cell transport control by ABA. To pinpoint the role of ABI1, the abi1-1 dominant mutant allele was stably transformed into Nicotiana benthamiana and its influence on IK,in, IK,out, and the anion channels was monitored in guard cells under voltage clamp. Compared with guard cells from wild-type and vector-transformed control plants, expression of the abi1-1 gene was associated with 2- to 6-fold reductions in IK,out and an insensitivity of both IK,in and IK,out to 20 microM ABA. In contrast, no differences between control and abi1-1 transgenic plants were observed in the anion current or its response to ABA. Parallel measurements of intracellular pH (pHi) using the fluorescent dye 2',7'-bis(2-carboxyethyl)-5-(and -6)-carboxyfluorescein (BCECF) in every case showed a 0.15- to 0.2-pH-unit alkalinization in ABA, demonstrating that the transgene was without effect on the pHi signal that mediates in ABA-evoked K+ channel control. In guard cells from the abi1-1 transformants, normal sensitivity of both K+ channels to and stomatal closure in ABA was recovered in the presence of 100 microM H7 and 0.5 microM staurosporine, both broad-range protein kinase antagonists. These results demonstrate an aberrant K+ channel behavior--including channel insensitivity to ABA-dependent alkalinization of pHi--as a major consequence of abi1-1 action and implicate AB11 as part of a phosphatase/kinase pathway that modulates the sensitivity of guard-cell K+ channels to ABA-evoked signal cascades.

Abscisic Acid↗

Tuberculosis case-finding in Western Australia.

The records of a total of 497 patients notified to have active tuberculosis (TB) in Western Australia from 1986 to 1990 inclusive have been reviewed in relation to case-finding. Of these, 276 (55.5%) were Asian migrants, 92 (18.5%) Europeans and 110 (22.1%) Australians. The disease was confirmed bacteriologically in 247 (49.7%). The most common mode of diagnosis was symptomatic presentation (39.8%), followed by migrant health surveillance (36.8%), incidental finding (7.5%), regular follow-up (3.8%) and pre-employment chest radiography (1.8%). Contact examination in the non-Asians accounted for only 0.4% while diagnosis after death 2.6%. There was no statistical difference in the detection of TB disease and infection among contacts of index patients with diseases at different sites or of different bacteriological status. The results of the study justify the present strict surveillance over the Asian migrants but do not support the overcautious policy of the contact examination procedures in the non-Asians, for which changes are proposed. Twelve patients whose confirmed TB was diagnosed only at autopsy had a mean age of 75.4 (range 41-86) years. Six of them had miliary or disseminated TB, three co-existing serious medical conditions and none a past history of the disease. The pattern has been well described and clinicians should keep a high index of suspicion for TB in this group of patients.

Adult↗