Community-onset Staphylococcus aureus infections presenting to general practices in South-eastern Australia.

Community-acquired Staphylococcus aureus infections are a public health concern, yet little is known about infections that do not present to hospital. We identified community-onset S. aureus infections via specimens submitted to a community-based pathology service. Referring doctors confirmed eligibility and described infection site, severity and treatment. Isolates were characterized on antibiotic resistance, PFGE, MLST/SCCmec, and Panton-Valentine leukocidin (PVL), representing 106 community-onset infections; 34 non-multiresistant methicillin-resistant S. aureus (nmMRSA) (resistant to <3 non-ß-lactam antibiotics), 15 multiply antibiotic-resistant MRSA (mMRSA) and 57 methicillin-sensitive S. aureus (MSSA). Most (93%) were skin and soft tissue infections. PVL genes were carried by 42% of nmMRSA isolates [95% confidence interval (CI) 26-61] and 15% of MSSA (95% CI 8-28). PVL was associated with infections of the trunk, head or neck (56·4% vs. 24·3%, P=0·005) in younger patients (23 vs. 52 years, P<0·001), and with boils or abscesses (OR 8·67, 95% CI 2·9-26·2), suggesting underlying differences in exposure and/or pathogenesis.

Inducible resistance to clindamycin in Staphylococcus aureus: validation of Vitek-2 against CLSI D-test.

AIMS: Inducible resistance to clindamycin in Staphylococcus aureus is common but not easily identified by routine testing, and can result in treatment failure if not detected. The gold standard method is the D-test described by the Clinical and Laboratory Standards Institute (CLSI). The Vitek-2 AST-P612 card contains an 'inducible clindamycin resistance' (ICR) test. We aimed to determine the accuracy of the Vitek-2 ICR test compared to the D-test. METHODS: Isolates of erythromycin non-susceptible, clindamycin susceptible Staphylococcus aureus were identified. Routine antimicrobial susceptibility testing was performed using the Vitek-2 AST-P612 card, including the ICR test, and compared against the D-test. RESULTS: 217 isolates were obtained. All of the 191 isolates that were ICR positive were D-test positive. Of the 27 ICR negative isolates, 10 (37%) were D-test positive [9 methicillin-sensitive S. aureus (MSSA), 1 methicillin-resistant S. aureus (MRSA)]. This correlates with a specificity of 100%, sensitivity of 95%, positive predictive value of 100%, and negative predictive value of 72%. CONCLUSIONS: The ICR test is reliable in the presence of a positive result; however there is a false negative rate of approximately one in four. This will lead to susceptibility reporting errors, with potentially serious clinical implications. A negative ICR should be confirmed by CLSI D-test before reporting clindamycin as susceptible where the organism is not susceptible to erythromycin.

The risk of exotic and native plants as hosts for four pest thrips (Thysanoptera: Thripinae).

Interactions among insect pests, crops and weeds are well recognised. In fact, the elimination of weed hosts outside of the crop is a common practice to control many insect-vectored viruses. However, little is known about interactions among insect pests, crops and native vegetation, and whether native plants may be used to revegetate areas where weed hosts have been eliminated as part of horticultural management regimes. We used the Northern Adelaide Plains horticultural region (South Australia, Australia) as a model system to study the potential of various plant taxa in hosting four pest thrips (three exotic, one native; Frankliniella occidentalis, F. schultzei, Thrips tabaci and T. imaginis) when located adjacent to, and distant from, horticultural crops. Flower funnels were used for standardised sampling of thrips on flowers from 19 exotic weed and 12 native plant species, representing 13 and three families, respectively. Flowers were sampled monthly over a year, and statistical analyses were performed to identify significant determinants of probability of thrips occurrence and density. Plant family was found to significantly influence both measures for each thrips species. In addition, crop proximity influenced the probability of occurrence for the two Frankliniella species (but only influenced density of the key pest F. occidentalis), and season influenced density of all four pest thrips. All native plant species tested had a low likelihood of hosting the three exotic thrips species. Overall, results suggest that judicious choice of surrounding vegetation has potential to be an important component of integrated pest management (IPM) while increasing biodiversity conservation.

Preference by Vespula germanica (Hymenoptera: Vespidae) for processed meats: implications for toxic baiting.

The German yellowjacket, Vespula germanica (F.) (Hymenoptera: Vespidae), was introduced into Australia in 1959 and has established throughout southern Australia. In urban environments, V. germanica is frequently a nuisance pest at public gatherings and to homeowners. In native environments, it has the potential to pose a threat to native invertebrates. The current practice for controlling the wasps is nest destruction with pesticide. However, locating the nest(s) is not always practical or possible. Meat baits impregnated with an insecticide that foraging wasps cut and carry back to the nest offer a means of suppressing wasps where the nest sites are unknown. The success of meat baits depends on the attractiveness and acceptance of the meat to the wasp and the mode of action of the insecticide. Our objective was to determine wasp preference and acceptance of five processed meats: canned chicken or fish and freeze-dried chicken, fish, or kangaroo. We found that more wasps visited and took freeze-dried kangaroo and canned chicken than the other baits. Canned and freeze-dried fish were similarly preferred, and freeze-dried chicken was the least attractive and accepted by foraging wasps. Our findings demonstrate that wasps prefer some processed meats and hence take more loads back to the nest. By combining a suitable insecticide with a meat bait preferred by wasps, the likelihood of effective suppression of nuisance wasp populations should be increased.

Phenotypic and genetic influences on test-day measures of acetone concentration in milk.

The objectives of this study were to estimate heritability of acetone concentration in milk, based on monthly samples of milk obtained as part of a routine milk testing program, and to evaluate the feasibility of using such data in a genetic evaluation program for selection against ketosis incidence. Milk samples were collected from January to December of 1999 in herds enrolled in the Ontario Dairy Herd Improvement Association, and acetone concentration was measured using an inline chemical procedure. The original data included more than 50,000 records. Because ketosis is generally a problem during early lactation, only the single test with the fewest days in milk was retained. In addition, data were retained only for cows with pedigree information. The final data set included 10,375 records. Among these data, only 6.56% had detectable levels of acetone. Acetone data were log-transformed prior to statistical analysis. Simple ANOVA indicated that herd, parity number, days in milk, and month of test had significant effects on acetone concentration. Acetone levels increased with lactation number and were higher in early lactation. Three approaches were applied for genetic analysis. First, REML was used with a simple linear animal model. Then, a separate procedure used data augmentation and Gibbs Sampling to obtain continuously distributed underlying values for records with zero acetone concentration, and these data were analyzed with both an animal and sire model. Heritability of acetone concentration was less than 1% for all 3 analyses. Herd effects accounted for about 5% of the phenotypic variance. Low estimates of heritability were due either to low actual levels of genetic variance or inability to detect all of the genetic variance present, due to infrequent recording during the early part of the lactation. Genetic evaluation based on recording of acetone concentration on a monthly basis seems of little use as a selection tool to decrease incidence of ketosis.

Estimation of genetic parameters for concentrations of milk urea nitrogen.

The objective of this study was to use field data collected by dairy herd improvement programs to estimate genetic parameters for concentrations of milk urea nitrogen (MUN). Edited data were 36,074 test-day records of MUN and yields of milk, fat, and protein obtained from 6102 cows in Holstein herds in Ontario, Canada. Data were divided into three sets, for the first three lactations. Two analyses were performed on data from each lactation. The first procedure used ANOVA to estimate the significance of the effects of several environmental factors on MUN. Herd-test-day effects had the most significant impact on MUN. Effects of stage of lactation were also important, and MUN levels tended to increase from the time of peak yield until the end of lactation. The second analysis used a random regression model to estimate heritabilities and genetic correlations of MUN and the yield traits. Heritability estimates for MUN in lactations one, two, and three were 0.44, 0.59, and 0.48, respectively. Heritabilities for the yield traits were of a similar magnitude. Little relationship was observed between MUN and yield. Raw phenotypic correlations were all <0.10 (absolute value). Genetic correlations with production traits were close to zero in lactations one and three and only slightly positive in lactation two. The results indicate that selection on MUN is possible, but relationships between MUN and other economically important traits such as metabolic disease and fertility are needed.

Emerging technologies in health care and the patient encounter of the future.

In the last five years, the Web has changed the way information is disseminated, knowledge is gained, and health care is provided. Even though the health care industry has yet to fully realize the potential of the Internet, the Web is only the beginning. Several emerging technologies will be changing the rules again. With new forms of network connectivity and faster, new wireless technologies, the adoption rate of mobile, handheld computers in health care will grow feverishly. Intelligent, autonomous software agents that guide the patient through the continuum of care will extend the reach of health care providers to all places at all times. The author discusses these new concepts, how they might be applied in health care, and the doctor-patient health care transaction.

Conservative management of a transdiaphragmatic fistula.

Case reports of transdiaphragmatic fistulas connecting subphrenic collections and empyemas are uncommon. We report the rare complication of a fistulous connection between a subphrenic collection and the bronchial tree.

Group cognitive training in old age: paper-and-pencil tasks of processing speed

Psychometric models attribute cognitive aging to decline in executive fuctions: processing speed(PS) or PS plus coordination of operations, both related to theprefrontal lobes. In a first study, healthy aged subjets (mean age: 68.42 years, range: 59-81; mean formalschooling: 6.47 years, range: 2-15) were divided into two groups. Group 1 (n=9) trained six VP paper-and-pencil tasks (letter and symbol recognizing speed) for five 1-hour daily sessions. Group 2 (n=8) received the same amount of training in a face-name association task. Measures of working memory employed in the pre-and post-tests were digit span, word span sentence comprehension. Group 1 improved significantly both in VP for letters and in the sentence comprehension measure. Group 2 improvement only in the paired associates learning task. Training old individuals on processing speed tasks may thus improve their working memory performance. In second training study, it was possible to assess coordination-related measures of working memory in the pre-and post-tests. Group 3 trained VP and was composed of 15 subjects with mean a age of 68.33 years (SD=6.36) and a mean formal schooling of 7.00 years (SD=3.22). Group 4, wich trained paired associates, consisted of 17 individuals with a mean age of 66.42 years (SD=8.38), who had a mean schooling of 7.77 years (SD=3.68). Contrary to our expectations, only training Group 4 exhibited generalization effects related to coordinatio of operations in working memory. These results confirm only partially unifactorial models of working memory

The changing role of the health care chief information officer.

Information is the lifeblood of the health care organization. In the past, chief information officers were responsible for nothing else but assuring a constant flow of information. Today, they are being asked to do a great deal more. From E-business to E-health strategy, the chief information officer is the focal point of an organization's ability to leverage new technology.

Protein S alters the active site location of activated protein C above the membrane surface. A fluorescence resonance energy transfer study of topography.

The location of the active site of membrane-bound activated protein C (APC) relative to the phospholipid surface was determined both in the presence and absence of its cofactor, protein S, using fluorescence resonance energy transfer (FRET). APC was chemically modified to create the FRET donor species, Fl-FPR-APC, with a fluorescein dye (Fl) covalently attached to the active site via a D-Phe-Pro-Arg (FPR) tether and located in the active site near S4. FRET was observed when Fl-FPR-APC was titrated in the presence of Ca2+ ions with phosphatidylcholine/phosphatidylserine (4:1) vesicles containing the FRET acceptor, octadecylrhodamine (OR). Assuming a random orientation of transition dipoles (kappa2 = 2/3), the average distance of closest approach between the fluorescein in the active site of the membrane-bound APC and the OR at the membrane surface is 94 A. The same calcium-dependent distance was obtained for both small and large unilamellar vesicles and for vesicles that contained phosphatidylethanolamine. The active site of membrane-bound APC is therefore located far above the phospholipid surface. Upon addition of protein S, the efficiency of Fl-FPR-APC to OR energy transfer increased due to a protein S-dependent rotational and/or translational movement of the APC protease domain relative to the surface. If this movement were solely translational, then the average height of the fluorescein in the membrane-bound APC.protein S complex would be 84 A above the surface. The extent of Fl-FPR-APC to OR energy transfer was unaltered by the addition of thrombin-inactivated protein S. The protein S effect was also specific for APC, since the addition of protein S to similarly-labeled derivatives of factor Xa, factor IXa, or factor VIIa did not alter the locations of their active sites. This direct measurement demonstrates that the binding of the protein S cofactor to its cognate enzyme elicits a relocation of the active site of APC relative to the membrane surface and thereby provides a structural explanation for the recently observed protein S-dependent change in the site of factor Va cleavage by APC.

Ochratoxin A in wheat: certification of two reference materials.

The preparation of two wheat reference materials and the certification of their ochratoxin A content is described. The materials were prepared and certified within the European Commission. Measurements and Testing Programme (M&T). The first and second phases of this project, two intercomparisons of procedures for the determination of ochratoxin A in wheat, at levels of approximately 13 micrograms/kg, and 7 micrograms/kg, respectively have already been reported. This paper describes the work carried out in certification of the ochratoxin A content (mass fraction) of two wheat reference materials: a blank wheat CRM 471, and a contaminated wheat, CRM 472. These materials were prepared for use in the second intercomparison referred to above and reported previously. Reference material CRM 472 was prepared from naturally-contaminated wheat blended with the blank wheat (CRM 471). Details are given of the milling, blending and packaging procedure, and the checks to ensure homogeneity and stability of the material. The certification exercise was carried out by 15 laboratories using a variety of extraction and clean-up procedures, and the certified ochratoxin A content (mass fraction) of CRM 471 was < 0.6 microgram/kg. The value for CRM 472 was 8.2 micrograms/kg with an uncertainty of 1.0 microgram/kg.

Ochratoxin A in wheat: a second intercomparison of procedures.

The European Commission, Measurements and Testing Programme (BCR) has undertaken a project to improve methodology and to prepare certified reference materials for ochratoxin A determination. The first phase of this project, an intercomparison of procedures for the determination of ochratoxin A in wheat, at a content of approximately 13 micrograms/kg, has already been reported. The second intercomparison study, described in this paper, involved 26 European laboratories, from 11 countries, which analysed wheat naturally contaminated at a level of approximately 7 micrograms/kg, and a 'blank' wheat sample (ochratoxin A content < 0.2 microgram/kg). The participants used a variety of procedures which involved different extraction solvents and clean-up procedures. All laboratories used HPLC as the determinative step. Some laboratories also used immunoaffinity column clean-up in comparison with their normal method. Recoveries of the normal methods used by laboratories ranged from 58 to 114%; only three laboratories obtained recoveries outside the accepted range of 70 to 110%. Recoveries of the immunoaffinity column methods, using two sources of column, ranged from 58 to 114% for one and from 4 to 86% for the other. The between-laboratory reproducibility coefficient of variation for all results was 34% for the normal methods, and 34 and 42% for the two types of immunoaffinity columns. It was noted that, after the results were corrected for spike recovery, some laboratories became outliers owing to low spike recoveries. Further investigations of the spiking protocols used by each laboratory showed that the time left for evaporation of the spiking solvent was crucial to the recovery obtained.

Moraxella catarrhalis: pathogenic significance in respiratory tract infections treated by community practitioners.

We prospectively studied the pathogenic significance of Moraxella (Branhamella) catarrhalis isolated from 212 patients of community practitioners in Australia. This organism was most commonly isolated during winter and early spring, and 92% of isolates were beta-lactamase producers. On the basis of predetermined clinical and microbiological criteria, 42% of the isolates were definitely pathogenic, 7% were probably pathogenic, 21% were of indeterminate pathogenicity, and 30% were nonpathogenic. Factors associated with pathogenic significance included pneumonia or bronchitis (87% of patients), predisposing respiratory or systemic conditions (62%), isolation from sputum, and pure isolation. Thirty-six percent of patients were < 5 years old, but only 9% of isolates from these patients were pathogenic or probably pathogenic, a finding that reflects the fact that nasal-swab and nasopharyngeal-aspirate sampling is a common practice. Isolates from older patients were more likely to be pathogenically significant. An assessment of the pathogenic significance of M. catarrhalis isolated from a patient in a community practice should take into consideration factors such as the patient's age, clinical illness, and underlying conditions; the presence of other organisms; and the source of the isolate.

Ochratoxin A in cereals and the BCR-M&T-projects.

Reliable analytical procedures and certified reference materials are essential for the establishment and enforcement of tolerance levels for ochratoxin A in foods. The inadequacy of analytical procedures, together with the need for certified reference materials, led the Commission of the European Communities Community Bureau of Reference (BCR) to undertake a project to prepare suitable reference materials for ochratoxin A in wheat, in order to improve methodology and to harmonise agreement of results between member states. The first intercomparison study indicated problems in the analysis due to the influence of co-extractives in the matrix, and demonstrated that further work was necessary to improve recovery, clean-up and reproducibility. The second intercomparison study, in the EC Measurements and Testing Programme, correlated the performance of the different methods for ochratoxin A measurement in a separate batch of contaminated wheat, and compared novel immunoaffinity column methods with the standard laboratory methods. Results were obtained from 26 laboratories within 11 European countries, which therefore gives a good representation of the scope of methods currently used in Europe. Considerable improvements in the determination of ochratoxin A were noted compared with the first intercomparison study.

Characteristics of HPRT-mutant T cell lines in a lupus patient treated with cyclophosphamide.

This report describes T cell lines derived from a patient with subacute cutaneous lupus after treatment with intravenous pulse cyclophosphamide. We selected for mitotically active, hypoxanthine-guanine phosphoribosyltransferase-deficient (HPRT-) T cells, by culture in a selective medium containing 6-thioguanine. When HPRT- cell lines were derived 6 days after pulse cyclophosphamide (CYC) treatment, they were predominantly CD8+ and T cell receptor (TCR) gamma/delta+, producing interferon-gamma (IFN gamma). Cell lines derived 21 days after CYC treatment were CD4+, TCR alpha/beta+ and produced both IFN gamma and interleukin-4. These results support a possible role for gamma/delta+ T cells in subacute cutaneous lupus and suggest a mechanism for the therapeutic effect of CYC.

Ochratoxin A in wheat: an intercomparison of procedures.

The Commission of the European Communities' Community Bureau of Reference (BCR) has undertaken a project to improve methodology and to prepare suitable certified reference materials in order to provide a basis for analytical quality control for the determination of ochratoxin A. The first phase of the project, an intercomparison of procedures for the determination of ochratoxin A in wheat at a level of approximately 13 micrograms/kg, is described. The study involved 24 European laboratories which analysed a naturally contaminated wheat and a 'blank' wheat sample (ochratoxin A content < 1 microgram/kg). The participants used a variety of procedures, including chloroform, methanol, toluene and ethyl acetate for extraction, and silica-, reversed phase- and immunoaffinity columns for clean-up. HPLC (one laboratory used TLC) was applied as the determinative step. Several performance characteristics were checked and the ochratoxin A content was determined. Recoveries were found to range from 25 to 100%. The coefficient of variation from all the results calculated on the basis of peak height was 23%. The study showed that the variation of results was influenced more by the clean-up step than by the extraction solvent. Some laboratories suffered significant day-to-day effects while others found difficulties with interfering peaks in the 'blank' material. It is planned for the next study to improve the recovery range, the clean-up step and the reproducibility (within-laboratory, between-days) and to check the influence of co-extractives from the matrix.

Clinical features of human infection with Scedosporium inflatum.

We report on 17 Australian cases of human infection or colonization with Scedosporium inflatum. The spectrum of clinical manifestations was similar to that in infection caused by Scedosporium apiospermum. The patients were classified into three groups. Four immunocompetent patients who presented with localized infections of a joint, nail bed, eye, or sphenoidal sinus made up the first group. Our first case, in a boy with posttraumatic septic arthritis, responded to surgical drainage with amphotericin B followed by treatment with itraconazole. The other three cases were cured by surgery alone. The second group consisted of five immunocompromised patients who presented with disseminated infections in a variety of sites. Four of these patients did not respond to antifungal therapy and died. The fifth apparently responded to antifungal drugs after correction of his neutropenia. The third group included eight patients with asymptomatic colonization in the external ear (five cases) or respiratory secretions (three cases). The nine isolates of S. inflatum tested by both disk and agar dilution methods were resistant to antifungal drugs. In our first case, which responded clinically to itraconazole, the MIC of this drug for the fungal isolate was 25 micrograms/mL. Thus S. inflatum can cause a broad spectrum of human infections whose severity and prognosis depend largely on the host's immune status.

Studies of bone marrow progenitor cells in lupus-prone mice. I. NZB marrow cells demonstrate increased growth in Whitlock-Witte culture and increased splenic colony-forming unit activity in the Thy-1-, lineage- population.

Previous studies have demonstrated that NZB marrow can transfer features of autoimmunity. Therefore, we undertook a study of NZB marrow to determine whether it demonstrated any phenotypic abnormalities. In Whitlock-Witte cultures, NZB marrow cells generated nonadherent cells at low seeding densities, densities at which marrow from other strains did not generate nonadherent cells. In contrast, NZB marrow grew less well than controls in Dexter cultures. Inasmuch as the latter favor growth of granulocyte-macrophage precursors and the former B cells, these results suggest a possible skewing of NZB marrow cells toward lymphocyte production. Unfractionated marrow cells from NZB mice were found to produce 10-fold more splenic colonies in lethally irradiated recipients than marrow cells from control mice. This result was independent of the genotype of the recipient. When the progenitor Thy-1lo, Lin- marrow subpopulation was studied, NZB mice did not differ substantially from controls regarding splenic CFU. Therefore, Thy-1-, Lin- marrow cells were studied as a possible source of the excess splenic CFU in NZB mice. Indeed, the NZB Thy-1-, Lin- population contained 30-fold more splenic CFU than did the Thy-1-, Lin- population from control mice. These results suggest that NZB mice have unusual marrow progenitor cells; such cells may play a role in their autoimmune disease.