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Aim: Rapid detection of biological agents in biodefense is critical for operational, tactical and strategic levels as well as for medical countermeasures. Yersinia pestis, Francisella tularensis, and Bacillus anthracis are high priority agents of biological warfare or bioterrorism and many response forces use lateral flow assays (LFAs) for their detection. Several companies produce these assays, which offer results in short time and are easy to use. Despite their importance, only few publications on the limits of detection (LOD) for LFAs are available. Most of these studies used inactivated bacteria or risk group‐2 strains. As the inactivation process in previous studies might have affected the tests' performances, it was our aim in this study to determine and compare the LOD of several commercially available LFAs using viable risk group‐3 strains. Methods and Results: Lateral flow assays from four different companies for the detection of following bacteria were evaluated: Y. pestis, F. tularensis and B. anthracis spores. Two independent quantification methods for each target organism were applied, in order to ensure high quantification accuracy. LODs varied greatly between tests and organisms and ranged between 104 for Y. pestis‐tests and as high as >109 for one B. anthracis‐test. Conclusion: This work precisely determined the LODs of LFAs from four commercial suppliers. The herein determined LODs differed from results of previous studies. This illustrates the need for using accurately quantified viable risk group 3‐strains for determining such LODs. Significance and Impact of the Study: Our work bridges an important knowledge gap with regard to LFA LOD. The LODs determined in this study will facilitate better assessment of LFA‐results. They illustrate that a negative LFA result is not suited to exclude the presence of the respective agent in the analyzed sample. [ABSTRACT FROM AUTHOR]

Bispecific antibodies (bsAb) are considered as promising tools for the elimination of disseminated tumour cells in a minimal residual disease situation. The bsAb-mediated recruitment of an immune effector cell in close vicinity of a tumour cell is thought to induce an antitumoural immune response. However, classical bispecific molecules activate only a single class of immune effector cell that may not yield optimal immune responses. We therefore constructed an intact bispecific antibody, BiUII (anti-CD3 × anti-EpCAM), that not only recognizes tumour cells and T lymphocytes with its two binding arms, but also binds and activates Fcγ-receptor positive accessory cells through its Fc-region. We have demonstrated recently that activated accessory cells contribute to the bsAb-induced antitumoural activity. We now analyse this stimulation in more detail and demonstrate here the BiUll-induced upregulation of activation markers like CD83 and CD95 on accessory cells and the induction of neopterin and biopterin synthesis. Experiments with pure cell subpopulations revealed binding of BiUll to CD64+ accessory cells and CD16+ NK cells, but not to CD32+ B lymphocytes. We provide further evidence for the importance of the Fc-region in that this bispecific molecule stimulates Fcγ-R-positive accessory cells to eliminate tumour cells in vitro by direct phagocytosis. [ABSTRACT FROM AUTHOR]

Irradiation of fully developed green leaves as well as irradiation of isolated chloroplasts causes a rapid, reversible increase in the activity of NADP-linked phosphoglyceraldehyde dehydrogenase. This increase is closely connected with the functioning of the non-cyclic electron flow of photosynthesis. It is accompanied by a decrease in the activity of the NAD-linked enzyme. The experiments presented here show that it is not the kinase but the dehydrogenase which is the rate limiting factor in the over-all reaction measured up to date. The activation of the ‘dark-enzyme’ (= basic enzyme) resulting in the ‘light-enzyme’ (= activated enzyme) is not due to an unspecific reduction of S-S-groups in the reactive center. Dithiothreitol only restores the activity lost during the isolation procedure of the chloroplasts. NADPH rather than dithiothreitol causes a specific activation of the basic enzyme. How- ever the basic enzyme must be present in the completely reduced form to undergo maximal activation. Incubation with dithiothreitol and NADPH thus results in an activation rate as high as that caused by light in vivo. Activation of the NADP-linked dehydrogenase could also be attained by NADPH after its partial purification by ammonium sulfate fractionation. The activation rate by incubation with different concentrations of NADPH shows a sigmoidal type of curve. There are characteristic differences in sensitivity between the activity of the basic enzyme and its ability to be activated at different temperatures between 52°–60°. The results indicate that the action of NADPH may be an anosteric one, causing a transition from a NAD- (or possibly a NAD + NADP)-dependent to a NADP-dependent form of the enzyme. A diagram for a feed-forward activation of the NADP-linked dehydrogenase which possibly takes part in regulation of C02-fixation is presented. [ABSTRACT FROM AUTHOR]

We have isolated genomic clones that contained the gene for human GTP cyclohydrolase I. One clone containing the 5'-regulatory region of this gene was further analysed. It encompassed the first exon, parts of the first intron and about 2.6kb of the promoter region. The transcription start site was localised by rapid amplification of cDNA ends (5'-RACE). The 2.6 kb region upstream of the transcription start site showed promoter activity when ligated upstream of a reporter gene. 5'-truncations of the promoter region increased its activity as long as the CAAT and TATA boxes remained unchanged. Several putative Spl responsive elements were located within a GC-rich region close to the transcription start site.

Damage-associated molecular patterns (DAMPs) and pathogen-associated molecular patterns (PAMPs) are key triggers of inflammation in sepsis. However, they have rarely been studied simultaneously. Thus, in the present study of patients with bacteraemic infection, we aimed to study how DAMP dynamics are linked to disease severity and outcome and to compare diagnostic and prognostic properties of a DAMP and a previously analysed PAMP (16S rDNA). In a prospective study of adult patients hospitalized with culture-proven community-onset bacteraemic infection, caused by Streptococcus pneumonia (n = 30), Staphylococcus aureus (n = 27), or Escherichia coli (n = 26), dynamics of a PAMP, i.e. 16S rDNA, have previously been presented. For the present study, blood samples obtained on hospital days 1-2 (when blood culture was positive), 3-4, 7 ± 1, 14 ± 2, and 28 ± 4 were analysed for four different DAMPs, i.e., nuclear DNA (nDNA), mitochondrial DNA (mtDNA), heat shock protein 90 alpha (HSP90α), and extracellular high mobility group box 1 (HMGB1). Sepsis was defined according to the Sepsis-3 criteria. The study outcomes were sepsis at admission and negative outcome, defined as intensive care unit (ICU) admission and/or death within 60 days. Of 83 study patients, sepsis was noted in 41 patients (49%) and a negative outcome was noted in 17 patients (20%). nDNA had areas under the receiver operating characteristic (ROC) curves of 0.78 for sepsis and 0.76 for negative outcome, which were higher than those of the other DAMPs and additional biomarkers (CRP, IL-6, IL-8, and IL-10). The nDNA and positive 16S rDNA results on day 1-2 were correlated with each other (r = 0.68, p < 0.001). Multivariate analyses showed that high day 1-2 concentrations of both nDNA and 16S rDNA were independently associated with sepsis. In addition, high day 1-2 concentration of nDNA was independently associated with negative outcomes. While 16S rDNA dissipated from the circulation within days, nDNA concentrations remained elevated throughout the follow-up period in patients with negative outcome. In conclusion, nDNA outperformed the other DAMPs regarding sepsis detection and outcome prediction. Both nDNA (a DAMP) and 16S rDNA (a PAMP) were independently linked to sepsis; nDNA was also associated with negative outcomes and persisted elevated in such cases. This highlights nDNA as an interesting marker within sepsis pathogenesis and as a promising clinical biomarker, warranting further studies., (© 2024. The Author(s).)

In December 2016 the clinical operation has started at the particle therapy centre MedAustron, Wiener Neustadt, Austria. Different commercial immobilisation devices are used for head patients. These immobilisation devices are a combination of table tops (Qfix BoS™ Headframe, Elekta HeadStep™), pillows (BoS™ Standard pillow, Moldcare®, HeadStep™ pillow) and thermoplastic masks (Klarity Green™, Qfix Fibreplast™, HeadStep™ iCAST double). For each patient image-guided radiotherapy (IGRT) is performed by acquiring orthogonal X-ray imaging and 2D3D registration and the application of the resulting 6-degree of freedom (DOF) position correction on the robotic couch. The inter- and intrafraction displacement of 101 adult head patients and 27 paediatric sedated head patients were evaluated and compared among each other regarding reproducibility during the entire treatment and stability during each fraction. For the comparison, statistical methods (Shapiro-Wilk test, Mann-Whitney U-test) were applied on the position corrections as well as on the position verifications. The actual planning target volume margins of 3mm (adults) and 2mm (children) were evaluated by applying the van Herk formula on the intrafraction displacement results and performing treatment plan robustness simulations of twelve different translational offset scenarios including a HU uncertainty of 3.5%. Statistically significant differences between the immobilisation devices were found, but they turned out to be clinically irrelevant. The margin calculation for adult head patients resulted in 0.8mm (lateral), 1.2mm (cranio-caudal) and 0.6mm (anterior-posterior), and for paediatric head patients under anaesthesia in 0.8mm (lateral), 0.5mm (cranio-caudal) and 0.9mm (anterior-posterior). Based on these values, robustness evaluations of selected adult head patients and sedated children showed the validity of the currently used PTV margins., (Copyright © 2021. Published by Elsevier GmbH.)

Background: Intraoperative radiotherapy with electrons (IOERT) boost could be not inferior to external beam radiotherapy (EBRT) boost in terms of local control and tissue tolerance. The aim of the study is to present the long-term follow-up results on local control, esthetic evaluation, and toxicity of a prospective study on early-stage breast cancer patients treated with breast-conserving surgery with an IOERT boost of 10 Gy (experimental group) versus 5 × 2 Gy EBRT boost (standard arm). Both arms received whole-breast irradiation (WBI) with 50 Gy (2 Gy single dose)., Methods: A single-institution phase III randomized study to compare IOERT versus EBRT boost in early-stage breast cancer was conducted as a non-inferiority trial. Primary endpoints were the evaluation of in-breast true recurrences (IBTR) and out-field local recurrences (LR) as well as toxicity and cosmetic results. Secondary endpoints were overall survival (OS), disease-free survival (DFS), and patient's grade of satisfaction with cosmetic outcomes., Results: Between 1999 and 2004, 245 patients were randomized: 133 for IOERT and 112 for EBRT. The median follow-up was 12 years (range 10-16 years). The cumulative risk of IBTR at 5-10 years was 0.8% and 4.3% after IOERT, compared to 4.2% and 5.3% after EBRT boost (p = 0.709). The cumulative risk of out-field LR at 5-10 years was 4.7% and 7.9% for IOERT versus 5.2% and 10.3% for EBRT (p = 0.762). All of the IOERT arm recurrences were observed at > 100 months' follow-up, whereas the mean time to recurrence in the EBRT group was earlier (55.2 months) (p < 0.05). No late complications associated with IOERT were observed. The overall cosmetic results were scored as good or excellent in physician and patient evaluations for both IOERT and EBRT. There were significantly better scores for IOERT at all time points in physician and patient evaluations with the greatest difference at the end of EBRT (p = 0.006 objective and p = 0.0004 subjective) and most narrow difference at 12 months after the end of EBRT (p = 0.08 objective and p = 0.04 subjective analysis)., Conclusion: A 10-Gy IOERT boost during breast-conserving surgery provides high local control rates without significant morbidity. Although not significantly superior to external beam boosts, the median time to local recurrences after IOERT is prolonged by more than 4 years.

Background: Blood stream infection (BSI) and sepsis are serious clinical conditions and identification of the disease-causing pathogen is important for patient management. The RISE (Rapid Identification of SEpsis) study was carried out to collect a cohort allowing high-quality studies on different aspects of BSI and sepsis. The aim of this study was to identify patients at high risk for BSI who might benefit most from new, faster, etiological testing using neutrophil to lymphocyte count ratio (NLCR) and Shapiro score., Methods: Adult patients (≥ 18 years) presenting at the emergency department (ED) with suspected BSI were prospectively included between 2014 and 2016 at Örebro University Hospital. Besides extra blood sampling, all study patients were treated according to ED routines. Electronic patient charts were retrospectively reviewed. A modified Shapiro score (MSS) and NLCR were extracted and compiled. Continuous score variables were analysed with area under receiver operator characteristics curves (AUC) to evaluate the ability of BSI prediction., Results: The final cohort consisted of 484 patients where 84 (17%) had positive blood culture judged clinically significant. At optimal cut-offs, MSS (≥3 points) and NLCR (> 12) showed equal ability to predict BSI in the whole cohort (AUC 0.71/0.74; sensitivity 69%/67%; specificity 64%/68% respectively) and in a subgroup of 155 patients fulfilling Sepsis-3 criteria (AUC 0.71/0.66; sensitivity 81%/65%; specificity 46%/57% respectively). In BSI cases only predicted by NLCR> 12 the abundance of Gram-negative to Gram-positive pathogens (n = 13 to n = 4) differed significantly from those only predicted by MSS ≥3 p (n = 7 to n = 12 respectively) (p < 0.05)., Conclusions: MSS and NLCR predicted BSI in the RISE cohort with similar cut-offs as shown in previous studies. Combining the MSS and NLCR did not increase the predictive performance. Differences in BSI prediction between MSS and NLCR regarding etiology need further evaluation.

Catalytic depolymerization of lignin is a challenging process due to competitive repolymerization reactions. In this paper, the oxidative depolymerization of lignin was catalyzed by a commercial laccase both in a batch experiment and in a membrane bioreactor using the same catalytic conditions. The membrane bioreactor was previously optimized to reach high permeation flux (25 L.h -1 .m -2 ) during lignin diafiltration. While the lignin was exclusively condensed in the batch experiment leading to high molecular weight macromolecules (from 9 to 16 kDa), its depolymerization was effective in the bioreactor producing fragments of less than 1 kDa thanks to the in-situ extraction of the reaction products. This paper demonstrates that the reactor configuration is playing an essential role in triggering or preventing lignin condensation. It also reports the first proof-of-concept demonstrating that in-situ membrane extraction of the reactive fragments of lignin from the bulk medium can be useful against detrimental repolymerization reactions., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Firstly studied and pointed out because of their major role in wood recalcitrance to delignification, LCCs are gradually raising interest about their own valorization thanks to their various interesting properties. Considering their future industrial production, this work is focused on evaluating the impact of different downstream processes (DSP) on their structure including moist heat sterilization, simple effect evaporation and spray-drying. LCCs were recovered from the mill water of a thermomechanical plant by successive membrane filtration steps. Samples were taken after the different DSP and compared to a freeze-dried control. Global and structural characterizations given by elemental analysis, acid hydrolysis, pyrolysis coupled to gas-chromatography, SEC, 13 C and 2D-HSQC NMR showed that the LCCs structure is well preserved during all of these DSP. This paper demonstrates the feasibility of producing high quality industrial LCC grade for commercialization in integrated biorefineries., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)