The uncertain consequences of transferring bacterial strains between laboratories - rpoS instability as an example

Abstract Background Microbiological studies frequently involve exchanges of strains between laboratories and/or stock centers. The integrity of exchanged strains is vital for archival reasons and to ensure reproducible experimental results. For at least 50 years, one of the most common means of shipping bacteria was by inoculating bacterial samples in agar stabs. Long-term cultures in stabs exhibit genetic instabilities and one common instability is in rpoS. The sigma factor RpoS accumulates in response to several stresses and in the stationary phase. One consequence of RpoS accumulation is the competition with the vegetative sigma factor σ70. Under nutrient limiting conditions mutations in rpoS or in genes that regulate its expression tend to accumulate. Here, we investigate whether short-term storage and mailing of cultures in stabs results in genetic heterogeneity. Results We found that samples of the E. coli K-12 strain MC4100TF exchanged on three separate occasions by mail between our laboratories became heterogeneous. Reconstruction studies indicated that LB-stabs exhibited mutations previously found in GASP studies in stationary phase LB broth. At least 40% of reconstructed stocks and an equivalent proportion of actually mailed stock contained these mutations. Mutants with low RpoS levels emerged within 7 days of incubation in the stabs. Sequence analysis of ten of these segregants revealed that they harboured each of three different rpoS mutations. These mutants displayed the classical phenotypes of bacteria lacking rpoS. The genetic stability of MC4100TF was also tested in filter disks embedded in glycerol. Under these conditions, GASP mutants emerge only after a 3-week period. We also confirm that the intrinsic high RpoS level in MC4100TF is mainly due to the presence of an IS1 insertion in rssB. Conclusions Given that many E. coli strains contain high RpoS levels similar to MC4100TF, the integrity of such strains during transfers and storage is questionable. Variations in important collections may be due to storage-transfer related issues. These results raise important questions on the integrity of bacterial archives and transferred strains, explain variation like in the ECOR collection between laboratories and indicate a need for the development of better methods of strain transfer.

Towards new botanical pesticides: the toxic effect of Eremanthus goyazensis (Asteraceae) leaves essential oil against Brevipalpus phoenicis (Acari: Tenuipalpidae)

This work reports the chemical characterization of Eremanthusgoyzensis essential oil and its toxic effect over Brevipalpus phoenicis. The essential oil displayed a major composition of sesquiterpenes (61.87%) including trans-caryophillene (26.81%) and germacrene-D (13.31%). The fumigation test indicated a promising bioactivity over adult B. phoenicis individuals at 24 h (2.03 µL/L of air) and 48 h (1.08 µL/L of air) of exposition. A brief discussion of essential oils composition and their singular role on the toxic effect over B. phoenicis is provided here. Our results may contribute to a new and profitable use of a species of Brazilian flora on agribusiness.

Variability in urinary oxalate measurements between six international laboratories

Hyperoxaluria is a major risk factor for kidney stone formation. Although urinary oxalate measurement is part of all basic stone risk assessment, there is no standardized method for this measurement.

Rivaroxaban: Quantification by anti-FXa assay and influence on coagulation tests A study in 9 Swiss laboratories

INTRODUCTION: Rivaroxaban (RXA) is licensed for prophylaxis of venous thromboembolism after major orthopaedic surgery of the lower limbs. Currently, no test to quantify RXA in plasma has been validated in an inter-laboratory setting. Our study had three aims: to assess i) the feasibility of RXA quantification with a commercial anti-FXa assay, ii) its accuracy and precision in an inter-laboratory setting, and iii) the influence of 10mg of RXA on routine coagulation tests. METHODS: The same chromogenic anti-FXa assay (Hyphen BioMed) was used in all participating laboratories. RXA calibrators and sets of blinded probes (aim ii.) were prepared in vitro by spiking normal plasma. The precise RXA content was assessed by high-pressure liquid chromatography-tandem mass spectrometry. For ex-vivo studies (aim iii), plasma samples from 20 healthy volunteers taken before and 2 - 3hours after ingestion of 10mg of RXA were analyzed by participating laboratories. RESULTS: RXA can be assayed chromogenically. Among the participating laboratories, the mean accuracy and the mean coefficient of variation for precision of RXA quantification were 7.0% and 8.8%, respectively. Mean RXA concentration was 114±43?g/L .RXA significantly altered prothrombin time, activated partial thromboplastin time, factor analysis for intrinsic and extrinsic factors. Determinations of thrombin time, fibrinogen, FXIII and D-Dimer levels were not affected. CONCLUSIONS: RXA plasma levels can be quantified accurately and precisely by a chromogenic anti-FXa assay on different coagulometers in different laboratories. Ingestion of 10mg RXA results in significant alterations of both PT- and aPTT-based coagulation assays.

Integrated engineering workstations in electrical engineering laboratories

This paper provides a description of integrated engineering workstations (IEW’s) used in undergraduate electrical engineering laboratories. The IEW’s are used for the design, analysis, and testing of engineering systems. Examples of laboratory experiments and software programs are presented.

Botanical drugs, synergy, and network pharmacology: forth and back to intelligent mixtures

For centuries the science of pharmacognosy has dominated rational drug development until it was gradually substituted by target-based drug discovery in the last fifty years. Pharmacognosy stems from the different systems of traditional herbal medicine and its "reverse pharmacology" approach has led to the discovery of numerous pharmacologically active molecules and drug leads for humankind. But do botanical drugs also provide effective mixtures? Nature has evolved distinct strategies to modulate biological processes, either by selectively targeting biological macromolecules or by creating molecular promiscuity or polypharmacology (one molecule binds to different targets). Widely claimed to be superior over monosubstances, mixtures of bioactive compounds in botanical drugs allegedly exert synergistic therapeutic effects. Despite evolutionary clues to molecular synergism in nature, sound experimental data are still widely lacking to support this assumption. In this short review, the emerging concept of network pharmacology is highlighted, and the importance of studying ligand-target networks for botanical drugs is emphasized. Furthermore, problems associated with studying mixtures of molecules with distinctly different pharmacodynamic properties are addressed. It is concluded that a better understanding of the polypharmacology and potential network pharmacology of botanical drugs is fundamental in the ongoing rationalization of phytotherapy.

Design of removable partial dentures: a survey of dental laboratories in Greece

The aim of this study was to compare data on design and fabrication methods of removable partial dentures (RPDs) in two major cities in Greece. A questionnaire was sent to 150 randomly selected dental technicians. The participation rate was 79.3%. The anterior palatal strap, the lingual bar, and the Roach-type clasp arm designs were preferred. Half of the RPDs fabricated were retained using precision attachments. Differences between the two cities were observed in types of major maxillary connectors used, types of attachments and impression materials used, as well as the design of distal-extension RPDs. Postdoctoral education was found to have an impact on RPD fabrication. Despite the differences observed, design and fabrication of RPDs followed commonly used principles.

Transferability of geoscientific information from various sources (study sites, underground rock laboratories, natural analogues) to support safety cases for radioactive waste repositories in argillaceous formations

In studies related to deep geological disposal of radioactive waste, it is current practice to transfer external information (e.g. from other sites, from underground rock laboratories or from natural analogues) to safety cases for specific projects. Transferable information most commonly includes parameters, investigation techniques, process understanding, conceptual models and high-level conclusions on system behaviour. Prior to transfer, the basis of transferability needs to be established. In argillaceous rocks, the most relevant common feature is the microstructure of the rocks, essentially determined by the properties of clay–minerals. Examples are shown from the Swiss and French programmes how transfer of information was handled and justified. These examples illustrate how transferability depends on the stage of development of a repository safety case and highlight the need for adequate system understanding at all sites involved to support the transfer.

Variability between laboratories performing coagulation tests with identical platforms: a nationwide evaluation study.

BACKGROUND While the assessment of analytical precision within medical laboratories has received much attention in scientific enquiry, the degree of as well as the sources causing variation between them remains incompletely understood. In this study, we quantified the variance components when performing coagulation tests with identical analytical platforms in different laboratories and computed intraclass correlations coefficients (ICC) for each coagulation test. METHODS Data from eight laboratories measuring fibrinogen twice in twenty healthy subjects with one out of 3 different platforms and single measurements of prothrombin time (PT), and coagulation factors II, V, VII, VIII, IX, X, XI and XIII were analysed. By platform, the variance components of (i) the subjects, (ii) the laboratory and the technician and (iii) the total variance were obtained for fibrinogen as well as (i) and (iii) for the remaining factors using ANOVA. RESULTS The variability for fibrinogen measurements within a laboratory ranged from 0.02 to 0.04, the variability between laboratories ranged from 0.006 to 0.097. The ICC for fibrinogen ranged from 0.37 to 0.66 and from 0.19 to 0.80 for PT between the platforms. For the remaining factors the ICC's ranged from 0.04 (FII) to 0.93 (FVIII). CONCLUSIONS Variance components that could be attributed to technicians or laboratory procedures were substantial, led to disappointingly low intraclass correlation coefficients for several factors and were pronounced for some of the platforms. Our findings call for sustained efforts to raise the level of standardization of structures and procedures involved in the quantification of coagulation factors.

Non-naturalized alien plants receive fewer flower visits than naturalized and native plants in a Swiss botanical garden

Many animal-pollinated plant species have been introduced to non-native regions without their usual pollinators. Nevertheless, some of these alien species managed to establish reproducing naturalized populations, which might negatively affect native plants. Recent studies have shown that many naturalized alien species can readily attract native pollinators. However, it is not known whether alien species that have not established naturalized populations are less successful in attracting pollinators. Therefore, we tested whether flower-visitation rates are lower for non-naturalized aliens than for naturalized alien and native species. We conducted a comparative study on flower visitation of 185 native, 37 naturalized alien and 224 non-naturalized alien plant species in the Botanical Garden of Bern, Switzerland. Our phylogenetically corrected analyses showed that non-naturalized alien species received fewer flower visitors than both naturalized alien and native species. Native, naturalized alien and non-naturalized alien species were visited by similar flower-visitor communities. Furthermore, among the naturalized alien species, the ones with a broader distribution range in Switzerland received a more diverse set of flower visitors. Although it has been suggested that most alien plants can readily integrate into native plant–pollinator networks, we show evidence that the capacity to attract flower visitors in non-native regions is different for naturalized and non-naturalized alien plants. Therefore, we conclude that successful naturalization of alien plants may be related to flower visitation.

The use of special stains at two dermatopathology laboratories in East Africa.

BACKGROUND Histopathology is often essential to establish an accurate diagnosis. Pathology laboratories are scarce in most Sub-Saharan Africa where dermatopathology is a developing field. In resource-poor countries, most specimens are analyzed only after hematoxylin and eosin staining. The availability of special stains is very limited and restricted to only few centers. The aim of this study is to analyze the extent of dermatopathological cases which can be adequately diagnosed after hematoxylin and eosin alone. Secondly, to investigate which cases required further special stains. METHODS All skin specimens submitted to two University Hospitals (Tanzania and Kenya) were included in this study. All specimens were first analyzed with hematoxylin and eosin and a diagnosis established when possible. All cases in which an accurate diagnosis after hematoxylin and eosin only was not possible, were registered and evaluated after further special stains. RESULTS A total of 386 specimens were examined. A proper histopathologic diagnosis with hematoxylin and eosin alone was possible in 344 (89.1%) samples. In 45 (11.6%) cases, mostly skin infections, further special stains were necessary. CONCLUSION A proper histopathologic diagnosis was possible after hematoxylin and eosin alone in almost 90% of the specimens submitted to the two laboratories in Sub-Saharan Africa.

Illustrated Botanical Books, Fifteenth, Sixteenth, Seventeenth Centuries

No attempt has been made for a comprehensive coverage of the many outstanding illustrated botanical books for this three hundred year period. Rather the aim has been for a combination of interesting illustrators and famous botanists. Like the exhibit on fine bird books, these books on display have been taken from the Rare Books division of Special Collections.