Assessment of bacteria and archaea in metalworking fluids using massive parallel 16S rRNA gene tag sequencing.

Determination of the bacterial diversity in industry-based liquid in-use water-miscible metalworking fluid (MWF) samples was targeted by massive parallel multiplex DNA sequencing, either directly or upon pretreatment with propidium monoazide (PMA) that allows differentiation between intact and physically damaged cells. As MWFs provide a suitable basis of life for micro-organisms, the majority is preserved by biocides. 'Bio-concept' fluids on the other hand are bactericide free, which intentionally leads to substantial bacterial populations. Samples from both fluid types were chosen: A median of 51 operational taxonomic units at genera level (OTUs) were detected per sample, but only 13 were present at or above 1·0% of the total population in any PMA-treated sample analysed. As both fluid types were mainly dominated by Pseudomonas spp., we resolved this genus on the species level and found the Pseudomonas oleovorans/pseudoalcaligenes group to predominate. We also looked for archaea and detected Methanobrevibacter spp., albeit in <3% of all samples analysed. SIGNIFICANCE AND IMPACT OF THE STUDY: Water-miscible metalworking fluids provide a suitable base of life for micro-organisms, mainly bacteria and fungi. Earlier publications suggested that the diversity is rather low, but these studies were largely based on heterotrophic plate counts. This might have resulted in underestimation of population density and microbial diversity as some organisms might just refuse to grow. This study used high-throughput sequencing in the absence and presence of propidium monoazide to explore bacterial and archaeal presence in metalworking fluids. We established that diversity is low and bacterial populations are dominated by the genus Pseudomonas spp.

Precision sampling measurements using ac programmable Josephson voltage standards.

We have performed a variety of precision measurements by comparing ac and dc waveforms generated by two independent ac programmable Josephson voltage standard (ACPJVS) systems. The objective of these experiments was to demonstrate the effectiveness of using a sampling digital voltmeter to measure small differences between Josephson waveforms for frequencies up to 3.6 kHz. The low uncertainties that we obtained confirm the feasibility of using this differential sampling method for high accuracy comparisons between ACPJVS waveforms and signals from other sources.

Electrostatic modulation of the superfluid density in an ultrathin film.

By capacitively charging an underdoped ultrathin La2-xSrxCuO4 film with an electric field applied across a gate insulator with a high dielectric constant, relative changes of the areal superfluid density ns of unprecedented strength were observed in measurements of the film kinetic inductance. Although ns appears to be substantially reduced by disorder, the data provide, for the first time on the same sample, direct compelling evidence for the Uemura relation Tc proportional to ns(T=0) in the underdoped regime of copper-oxide superconductors.

[Percutaneous vertebroplasty: methods, indications, results]. / Perkután vertebroplasztika: módszer, indikációk, eredmények.

Percutaneous vertebroplasty (PVP) is a radiologically guided therapeutic procedure, which consists of percutaneous injection of a liquid polymer (bone cement) into a destroyed vertebral body. PVP was invented in 1984, in France, first for treating vertebral body haemangioma. Since its introduction the indications have been expanded progressively and today PVP is indicated mainly for treatment of vertebral haemangioma, malignant vertebral tumor and osteoporotic vertebral compression fracture. The unique advantage of this technique is that besides the stabilization of the vertebral body--and partly in connection with this--it affords prompt and lasting pain relief. Based on published data the success rate of the procedure is 80-100% with a complication rate of 1-10%. Thus, PVP is a valuable minimally invasive tool, providing immediate pain relief and early mobility in carefully selected patients. However, further work is needed to define the benefits of PVP compared to the standard treatment. The purpose of this paper is to demonstrate the technique by analyzing scientific reports published to date and summarizing the first author's own experience gained at the University Hospital of Geneva, Department of Neuroradiology, Switzerland.

Chloroplasts of the green alga Chlamydomonas reinhardtii possess at least four distinct stromal processing proteases.

The majority of the proteins in the chloroplast are encoded in the nucleus and synthesised in the cytoplasm as precursors with N-terminal extensions. These targeting sequences guide the precursor proteins into the chloroplast where they are immediately cleaved off by a stromal processing protease (SPP). It is commonly assumed that in higher plant chloroplasts one general SPP processes almost all imported precursor proteins. In the green alga Chlamydomonas, however, there exist several different SPPs which process the various Chlamydomonas precursor proteins. The seven precursor proteins investigated here, which were all correctly imported into isolated chloroplasts, could be divided into two groups: Four precursor proteins were cleaved correctly when processed in vitro with an extract of stromal proteins. Four different SPPs were found in Chlamydomonas chloroplasts to be responsible for the processing of this class of precursors and these four activities were separated chromatographically, characterised and further distinguished by their sensitivity to different inhibitors. The three precursors of the second group were degraded completely by unidentified enzyme(s) present in the stromal extract. Degradation of these precursors was dependent on their conformational integrity as well as on the redox state in the stroma.

Isolation of thylakoid membrane vesicles of Chlamydomonas reinhardii chloroplasts that are able to integrate and import in vitro synthesized precursor proteins.

Once imported into the stroma, nuclear encoded proteins of the chloroplast have to be routed to their final compartment, e.g. the thylakoid membranes. Four different pathways have been reported for the translocation of precursor proteins across and for the integration of mature proteins into the thylakoid membranes in higher plants. To study the sorting of precursor proteins in chloroplasts of higher plants the generation of an in vitro system using isolated intact thylakoid membrane vesicles was of major importance. Here we report the isolation of intact thylakoid membrane vesicles of the green algae Chlamydomonas reinhardii for the generation of a similar algal system. Further we show successful transport of several Chlamydomonas precursor proteins into isolated thylakoids: Lumenal precursors were translocated into the vesicles resulting in the accumulation of their mature, thermolysin-insensitive forms and thylakoid membrane proteins were specifically integrated into isolated Chlamydomonas thylakoid membranes.