Certification of a Reference Material Consisting of Genomic DNA Inserts of Bacillus Subtilis DSM 5750 for PFGE, IRMM-312

This report describes the production and certification of IRMM-312, a reference material of genomic DNA (gDNA) of Bacillus subtilis DSM 5750 in agarose inserts. This CRM (IRMM-312) is intended to be used for the taxonomic identification of authorised probiotic feed additives by Pulsed Field Gel Electrophoresis (PFGE). The batch was found to be homogeneous and the material is stable at 4 °C. The batch was characterised by five laboratories determining the PFGE pattern of a SfiI restriction enzyme digest of IRMM-312. The pattern should be considered as a whole and restriction fragments in the size interval from 15 kb to 97 kb are certified for their fragment length.JRC.D.2-Reference material

Certification of a Reference Material with Eschericia Coli O157 (NCTC 12900) at a level of 4 Colony forming Unit per Material Sphere, IRMM-351

This report describes the certification of a reference material (IRMM-351) of Escherichia coli O157. Certified Reference Materials (CRMs) for microbiological analysis are indispensable tools for development and validation of detection methods and for the implementation and support of internal and external quality control in the area of microbiological analysis. Each vial contains one material sphere of E. coli O157. The homogeneity and stability (at 4 °C, -20 °C and -70 °C) of the batch was assessed by monitoring colony forming units (cfu) on nutrient agr (NA) and eterohemolysin agar (EhlyA) of selected vials by colony counting. The material is not stable at 4 °C but no instability was detected when stored at -20 °C for up to 12 months and at -70 °C for up to 54 months. The batch was characterised by six laboratories to determine a certified value of cfu per vial on NA and EhlyA. The certified value is 4 cfu on both agars with an expanded uncertainty of 2 using a coverage factor k = 2, corresponding to a level of confidence of about 95 %. DNA sequence analysis of the coding region for the fliC gene identified the material as E. coli O157.JRC.D.2-Reference material

Certification of a Reference Material with Salmonella Enteritidis (NCTC 12694) at a Level of 5 Colony Forming Units on Nutrient Agar and 4 Colony Forming Units on Xylose Lysine Deoxycholate Agar, IRMM-352

This report describes the certification of a reference material (IRMM-352) of Salmonella Enteritidis. Certified Reference Materials (CRMs) for microbiological analysis are indispensable tools for development and validation of detection methods and for the implementation and support of internal and external quality control in the area of microbiological analysis. Each vial contains one material sphere of S. enteritidis. The homogeneity and stability (at 4 °C, -20 °C and -70 °C) of the batch was assessed by monitoring colony forming units (cfu) on nutrient agar (NA) and xylose lysine deoxycholate (XLD) agar of selected vials by colony counting. The material is not stable at 4 °C byt no instability was detected when stored at -20 °C for up to 12 months and at -70 °C for up to 54 months. The batch was characterised by six laboratories to determine a certified value of cfu per vial on NA and XLD agar. A quite considerable performance difference of both agars was observed. The certified value is 5 cfu on NA with an expanded uncertainty of 2 and 4 cfy on XLD agar with an expanded incertainty of 2 using a coverage factor k = 2, corresponding to a level of confidence of about 95 %. DNA sequence analysis of the coding region for the sefA gene identified the material as S. enteritidis.JRC.D.2-Reference material

Prediction of Phase Behavior of Spray-Dried Amorphous Solid Dispersions: Assessment of Thermodynamic Models, Standard Screening Methods and a Novel Atomization Screening Device with Regard to Prediction Accuracy

The evaluation of drug–polymer miscibility in the early phase of drug development is essential to ensure successful amorphous solid dispersion (ASD) manufacturing. This work investigates the comparison of thermodynamic models, conventional experimental screening methods (solvent casting, quench cooling), and a novel atomization screening device based on their ability to predict drug–polymer miscibility, solid state properties (Tg value and width), and adequate polymer selection during the development of spray-dried amorphous solid dispersions (SDASDs). Binary ASDs of four drugs and seven polymers were produced at 20:80, 40:60, 60:40, and 80:20 (w/w). Samples were systematically analyzed using modulated differential scanning calorimetry (mDSC) and X-ray powder diffraction (XRPD). Principal component analysis (PCA) was used to qualitatively assess the predictability of screening methods with regards to SDASD development. Poor correlation was found between theoretical models and experimentally-obtained results. Additionally, the limited ability of usual screening methods to predict the miscibility of SDASDs did not guarantee the appropriate selection of lead excipient for the manufacturing of robust SDASDs. Contrary to standard approaches, our novel screening device allowed the selection of optimal polymer and drug loading and established insight into the final properties and performance of SDASDs at an early stage, therefore enabling the optimization of the scaled-up late-stage development

A century of temporal stability of genetic diversity in wild bumblebees

Since the 1950s, bumblebee (Bombus) species are showing a clear decline worldwide. Although many plausible drivers have been hypothesized, the cause(s) of this phenomenon remain debated. Here, genetic diversity in recent versus historical populations of bumblebee species was investigated by selecting four currently restricted and four currently widespread species. Specimens from five locations in Belgium were genotyped at 16 microsatellite loci, comparing historical specimens (1913-1915) with recent ones (2013-2015). Surprisingly, our results showed temporal stability of genetic diversity in the restricted species. Furthermore, both historical and recent populations of restricted species showed a significantly lower genetic diversity than found in populations of co-occurring widespread species. The difference in genetic diversity between species was thus already present before the alleged recent drivers of bumblebee decline could have acted (from the 1950's). These results suggest that the alleged drivers are not directly linked with the genetic variation of currently declining bumblebee populations. A future sampling in the entire distribution range of these species will infer if the observed link between low genetic diversity and population distribution on the Belgium scale correlates with species decline on a global scale

Development of a small-scale spray-drying approach for amorphous solid dispersions (ASDs) screening in early drug development

The present study details the development of a small-scale spray-drying approach for the routine screening of amorphous solid dispersions (ASDs). This strategy aims to overcome the limitations of standard screening methodologies like solvent casting and quench cooling to predict drug-polymer miscibility of spraydried solid dispersions (SDSDs) and therefore to guarantee appropriate carrier and drug-loading (DL) selection. A DoE approach was conducted to optimize process conditions of ProCept 4M8-TriX spray-drying to maximize the yield from a 100 mg batch of Itraconazole/HPMCAS-LF and Itraconazole/Soluplus 40:60 (w/w). Optimized process parameters include: inlet temperature, pump speed, drying and atomizing airflows. Identified process conditions derived from the DoE analysis were further i) tested with Itraconazole, Naproxen and seven polymers, ii) adapted for small cyclone use, iii) downscaled to 20 mg batch production. Drug-polymer miscibility was systematically characterized using modulated differential scanning calorimetry (mDSC). Spray-drying was identified as a well-suited screening approach: mean yield of 10.1 to 40.6% and 51.1 to 81.0% were obtained for 20 and 100 mg ASD productions, respectively. Additionally, this work demonstrates the interest to move beyond conventional screening approaches and integrate spray-drying during screening phases so that a greater prediction accuracy in terms of SDSDs miscibility and performance can be obtained. Keywords: amorphous solid dispersions; spray-dryer; design of experiments; screening; miscibility; solvent casting; quench cooling; polymer

HD 172555: Detection of 63 μ m [OI] emission in a debris disc

Astronomy and Astrophysics 546 (2012): L8 Reproduced with permission from Astronomy & AstrophysicsContext. HD 172555 is a young A7 star belonging to the β Pictoris moving group that harbours a debris disc. The Spitzer/IRS spectrum of the source showed mid-IR features such as silicates and glassy silica species, indicating the presence of a warm dust component with small grains, which places HD 172555 among the small group of debris discs with such properties. The IRS spectrum also shows a possible emission of SiO gas. Aims. We aim to study the dust distribution in the circumstellar disc of HD 172555 and to asses the presence of gas in the debris disc. Methods. As part of the GASPS open time key programme, we obtained Herschel/PACS photometric and spectroscopic observations of the source.We analysed PACS observations of HD 172555 and modelled the spectral energy distribution with a modified blackbody and the gas emission with a two-level population model with no collisional de-excitation. Results. We report for the first time the detection of [OI] atomic gas emission at 63.18 μm in the HD 172555 circumstellar disc. We detect excesses due to circumstellar dust toward HD 172555 in the three photometric bands of PACS (70, 100, and 160 μm).We derive a large dust particle mass of (4.8 ± 0.6) × 10−4 M⊕ and an atomic oxygen mass of 2.5 × 10−2R2 M⊕, where R in AU is the separation between the star and the inner disc. Thus, most of the detected mass of the disc is in the gaseous phaseThis research has been funded by Spanish grants AYA 2010-21161-C02-02, CDS2006-00070 and PRICIT-S2009/ESP-1496. J.-C. Augereau and J. Lebreton thank the ANR (contract ANR-2010 BLAN-0505-01, EXOZODI) and the CNES-PNP for financial support. C. Pinte, F. Menard and W.-F. Thi acknowledges funding from the EU FP7-2011 under Grant Agreement nr. 284405. G. Meeus is supported by RYC-2011-07920. G. Meeus, C. Eiroa, I. Mendigutía and B. Montesinos are partly supported by AYA-2011-26202. F.M. acknowledges support from the Millennium Science Initiative (Chilean Ministry of Economy), through grant ÒNucleus P10-022-F

Short- and long-lasting tinnitus relief induced by transcranial direct current stimulation

A significant proportion of the population suffers from tinnitus, a bothersome auditory phantom perception that can severely alter the quality of life. Numerous experimental studies suggests that a maladaptive plasticity of the auditory and limbic cortical areas may underlie tinnitus. Accordingly, repetitive transcranial magnetic stimulation (rTMS) has been repeatedly used with success to reduce tinnitus intensity. The potential of transcranial direct current stimulation (tDCS), another promising method of noninvasive brain stimulation, to relieve tinnitus has not been explored systematically. In a double-blind, placebo-controlled and balanced order design, 20 patients suffering from chronic untreatable tinnitus were submitted to 20 minutes of 1 mA anodal, cathodal and sham tDCS targeting the left temporoparietal area. The primary outcome measure was a change in tinnitus intensity or discomfort assessed with a Visual Analogic Scale (VAS) change-scale immediately after tDCS and 1 hour later. Compared to sham tDCS, anodal tDCS significantly reduced tinnitus intensity immediately after stimulation; whereas cathodal tDCS failed to do so. The variances of the tinnitus intensity and discomfort VAS change-scales increased dramatically after anodal and cathodal tDCS, whereas they remained virtually unchanged after sham tDCS. Moreover, several patients unexpectedly reported longer-lasting effects (at least several days) such as tinnitus improvement, worsening, or changes in tinnitus features, more frequently after real than sham tDCS. Anodal tDCS is a promising therapeutic tool for modulating tinnitus perception. Moreover, both anodal and cathodal tDCS seem able to alter tinnitus perception and could, thus, be used to trigger plastic changes