Haemoptysis as the first presentation of COVID-19 : a case report

Background Coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is an ongoing pandemic that profoundly challenges healthcare systems all over the world. Fever, cough and fatigue are the most commonly reported clinical symptoms. Case presentation A 58-year-old man presented at the emergency department with acute onset haemoptysis. On the fifth day after admission, he developed massive haemoptysis. Computed tomography (CT) angiography of the chest revealed alveolar haemorrhage, more prominent in the left lung. Flexible bronchoscopy confirmed bleeding from the left upper lobe, confirmed by a bronchial arteriography, which was successfully embolized. Nasopharyngeal swabs (NPS) tested for SARS-CoV-2 using real-time polymerase chain reaction (RT-PCR) repeatedly returned negative. Surprisingly, SARS-CoV-2 was eventually detected in bronchoalveolar lavage (BAL) fluid. Conclusions Life-threatening haemoptysis is an unusual presentation of COVID-19, reflecting alveolar bleeding as a rare but possible complication. This case emphasises the added value of bronchoscopy with BAL in the diagnostic work-up in case of high clinical suspicion and negative serial NPS in patients presenting with severe symptoms

Assessment of Microbial Diversity in Biofilms Recovered from Endotracheal Tubes Using Culture Dependent and Independent Approaches

Ventilator-associated pneumonia (VAP) is a common nosocomial infection in mechanically ventilated patients. Biofilm formation is one of the mechanisms through which the endotracheal tube (ET) facilitates bacterial contamination of the lower airways. In the present study, we analyzed the composition of the ET biofilm flora by means of culture dependent and culture independent (16 S rRNA gene clone libraries and pyrosequencing) approaches. Overall, the microbial diversity was high and members of different phylogenetic lineages were detected (Actinobacteria, beta-Proteobacteria, Candida spp., Clostridia, epsilon-Proteobacteria, Firmicutes, Fusobacteria and gamma-Proteobacteria). Culture dependent analysis, based on the use of selective growth media and conventional microbiological tests, resulted in the identification of typical aerobic nosocomial pathogens which are known to play a role in the development of VAP, e.g. Staphylococcus aureus and Pseudomonas aeruginosa. Other opportunistic pathogens were also identified, including Staphylococcus epidermidis and Kocuria varians. In general, there was little correlation between the results obtained by sequencing 16 S rRNA gene clone libraries and by cultivation. Pyrosequencing of PCR amplified 16 S rRNA genes of four selected samples resulted in the identification of a much wider variety of bacteria. The results from the pyrosequencing analysis suggest that these four samples were dominated by members of the normal oral flora such as Prevotella spp., Peptostreptococcus spp. and lactic acid bacteria. A combination of methods is recommended to obtain a complete picture of the microbial diversity of the ET biofilm

TBX2 is a neuroblastoma core regulatory circuitry component enhancing MYCN/FOXM1 reactivation of DREAM targets

Chromosome 17q gains are almost invariably present in high-risk neuroblastoma cases. Here, we perform an integrative epigenomics search for dosage-sensitive transcription factors on 17q marked by H3K27ac defined super-enhancers and identify TBX2 as top candidate gene. We show that TBX2 is a constituent of the recently established core regulatory circuitry in neuroblastoma with features of a cell identity transcription factor, driving proliferation through activation of p21-DREAM repressed FOXM1 target genes. Combined MYCN/TBX2 knockdown enforces cell growth arrest suggesting that TBX2 enhances MYCN sustained activation of FOXM1 targets. Targeting transcriptional addiction by combined CDK7 and BET bromodomain inhibition shows synergistic effects on cell viability with strong repressive effects on CRC gene expression and p53 pathway response as well as several genes implicated in transcriptional regulation. In conclusion, we provide insight into the role of the TBX2 CRC gene in transcriptional dependency of neuroblastoma cells warranting clinical trials using BET and CDK7 inhibitors

Effect of anti-interleukin drugs in patients with COVID-19 and signs of cytokine release syndrome (COV-AID): a factorial, randomised, controlled trial.

BACKGROUND: Infections with SARS-CoV-2 continue to cause significant morbidity and mortality. Interleukin (IL)-1 and IL-6 blockade have been proposed as therapeutic strategies in COVID-19, but study outcomes have been conflicting. We sought to study whether blockade of the IL-6 or IL-1 pathway shortened the time to clinical improvement in patients with COVID-19, hypoxic respiratory failure, and signs of systemic cytokine release syndrome. METHODS: We did a prospective, multicentre, open-label, randomised, controlled trial, in hospitalised patients with COVID-19, hypoxia, and signs of a cytokine release syndrome across 16 hospitals in Belgium. Eligible patients had a proven diagnosis of COVID-19 with symptoms between 6 and 16 days, a ratio of the partial pressure of oxygen to the fraction of inspired oxygen (PaO(2):FiO(2)) of less than 350 mm Hg on room air or less than 280 mm Hg on supplemental oxygen, and signs of a cytokine release syndrome in their serum (either a single ferritin measurement of more than 2000 μg/L and immediately requiring high flow oxygen or mechanical ventilation, or a ferritin concentration of more than 1000 μg/L, which had been increasing over the previous 24 h, or lymphopenia below 800/mL with two of the following criteria: an increasing ferritin concentration of more than 700 μg/L, an increasing lactate dehydrogenase concentration of more than 300 international units per L, an increasing C-reactive protein concentration of more than 70 mg/L, or an increasing D-dimers concentration of more than 1000 ng/mL). The COV-AID trial has a 2 × 2 factorial design to evaluate IL-1 blockade versus no IL-1 blockade and IL-6 blockade versus no IL-6 blockade. Patients were randomly assigned by means of permuted block randomisation with varying block size and stratification by centre. In a first randomisation, patients were assigned to receive subcutaneous anakinra once daily (100 mg) for 28 days or until discharge, or to receive no IL-1 blockade (1:2). In a second randomisation step, patients were allocated to receive a single dose of siltuximab (11 mg/kg) intravenously, or a single dose of tocilizumab (8 mg/kg) intravenously, or to receive no IL-6 blockade (1:1:1). The primary outcome was the time to clinical improvement, defined as time from randomisation to an increase of at least two points on a 6-category ordinal scale or to discharge from hospital alive. The primary and supportive efficacy endpoints were assessed in the intention-to-treat population. Safety was assessed in the safety population. This study is registered online with ClinicalTrials.gov (NCT04330638) and EudraCT (2020-001500-41) and is complete. FINDINGS: Between April 4, and Dec 6, 2020, 342 patients were randomly assigned to IL-1 blockade (n=112) or no IL-1 blockade (n=230) and simultaneously randomly assigned to IL-6 blockade (n=227; 114 for tocilizumab and 113 for siltuximab) or no IL-6 blockade (n=115). Most patients were male (265 [77%] of 342), median age was 65 years (IQR 54-73), and median Systematic Organ Failure Assessment (SOFA) score at randomisation was 3 (2-4). All 342 patients were included in the primary intention-to-treat analysis. The estimated median time to clinical improvement was 12 days (95% CI 10-16) in the IL-1 blockade group versus 12 days (10-15) in the no IL-1 blockade group (hazard ratio [HR] 0·94 [95% CI 0·73-1·21]). For the IL-6 blockade group, the estimated median time to clinical improvement was 11 days (95% CI 10-16) versus 12 days (11-16) in the no IL-6 blockade group (HR 1·00 [0·78-1·29]). 55 patients died during the study, but no evidence for differences in mortality between treatment groups was found. The incidence of serious adverse events and serious infections was similar across study groups. INTERPRETATION: Drugs targeting IL-1 or IL-6 did not shorten the time to clinical improvement in this sample of patients with COVID-19, hypoxic respiratory failure, low SOFA score, and low baseline mortality risk. FUNDING: Belgian Health Care Knowledge Center and VIB Grand Challenges program

Een ongewone oorzaak van hemoptoë : pulmonaal-veneuze stenose na radiofrequentieablatie van voorkamerfibrillatie

Pulmonaal-veneuze stenose (PVS) is een zeldzame, maar ernstige complicatie na radiofrequentieablatie van voorkamerfibrillatie (VKF). De diagnose is vaak moeilijk door de aspecifieke presentatie en wordt dan ook gemakkelijk gemist. Een vroegtijdige behandeling met een ballonangioplastiek (BA), al dan niet in combinatie met een stentimplantatie, verbetert de klinische uitkomst en vermindert de kans op complicaties. In dit artikel wordt de ziektegeschiedenis beschreven van een 47-jarige patiënt met recidiverende hemoptoë en thoracale pijn als gevolg van PVS na radiofrequentieablatie, succesvol behandeld met een BA en de implantatie van een bare-metal stent (BMS)

Detection, separation and sequencing of oligosaccharides in beer

De volmondigheid van een bier wordt ondermeer bepaald door de oligosachariden van het restextact (1). Drie krachtige methoden werden gecombineerd om het complexe sacharideprofiel in bier weer te geven: HPAEC-PAD (High pH Anion Exchange Chromatography with Pulsed Amperometric detection), DSA-FACE (Fluorescent Assisted Carbohydrate Electrophoresis uitgevoerd op een capillaire array DNA Sequenator) en MALDI TOF MS (Matrix Assisted Laser Desorption/Ionisation with Time-of-Flight Mass Spectrometer). De sacharidesamenstelling van verschillende bieren werd met elkaar vergeleken en oligosachariden uit één type bier werden preparatief gescheiden aan de hand van Carbograph-SPE (Solid Phase Extraction) en gelfiltratie. De bekomen sachariden werden enzymatisch gesequeneerd. Met HPAEC-PAD worden sachariden gescheiden op basis van hun lading en structuur. De detectie gebeurt zonder voorafgaande derivatisering wat massaspectrometrische analyse van de sachariden toelaat na de scheiding. De detectiegrens voor malto-oligosachariden bedraagt 10 pmol en de resolutie voor de malto-oligosachariden in bier met DP<10 is zeer goed (Figuur 1, links). Carbohydraatelektroforese op DNA sequeneringsapparatuur laat toe om met zeer hoge gevoeligheid en resolutie glycaanmengsels te "fingerprinten". Deze analysetechniek werd oorspronkelijk geperfectioneerd voor complexe N-glycaanmengsels (2) en werd in dit werk toegepast op de oligosachariden in bier. De suikers worden vooraf gederivatiseerd met het fluorescent label APTS (9-aminopyrene-1,4,6-trisulfonaat). De detectiegrens voor de malto-oligosachariden bedraagt 10 fmol en de resolutie blijft bewaard met stijgende grootte van de sachariden (Figuur 2, rechts). Met MALDI-TOF MS wordt de massa van de oligosachariden bepaald zonder risico tot fragmentatie. De onderzochte bieren verschillen sterk in concentraties aan hogere malto-oligosachariden en isomalto-oligosachariden (Figuur 1). Sommige bieren bevatten veel α(1→4) gebonden en weinig α(1→6) vertakte sachariden, voor andere geldt het omgekeerde. De profielen vertonen niet echt een tendens per type bier. Deze verschillen kunnen hun weerslag hebben op de organoleptische eigenschappen van het bier en in het bijzonder op de volmondigheid. Uit één type bier dat veel α(1→6) vertakte sachariden bevat, werden de oligosachariden met een DP hoger dan 5 preparatief gescheiden op basis van hun moleculair gewicht. De oligosachariden werden geadsorbeerd op koolstof gepakt in een solid phase extraction kolom en geëlueerd met een stapgradiënt van water en n-butanol gesatureerd water (3). De oligosachariden elueerden in stijgende grootte en werden geïsoleerd in batch. De fractie geëlueerd met 1% butanol bestaat bijvoorbeeld uit vier isomalto-oligosachariden. MALDI-TOF MS analyse wijst op een mengsel van een hexa-, hepta-, octa- en nonasachariden. Deze vier oligosachariden elueren als enkelvoudige pieken voor de α(1→4) gebonden malto-oligosachariden (M6-M9) bij HPAEC-PAD analyse. DSA-FACE analyse toont echter dat iedere piek bestaat uit verschillende oligosachariden, waarschijnlijk positie-isomeren met dezelfde DP (Figuur 2). De oligosachariden werden verder gescheiden met Biogel P4 gelfiltratie chromatografie en de scheiding werd opnieuw gecontroleerd met HPAEC-PAD. De afzonderlijke oligosachariden werden enzymatisch gesequeneerd: specifieke enzymen werden gebruikt om de structuur van de oligosachariden op te helderen. De gevormde producten werden geanalyseerd met HPAEC-PAD en DSA-FACE. Zo werd een isomalto-oligosacharide met DP9 (drie pieken op DSA-FACE) na een pullulanase-behandeling enkel omgezet tot maltohexaose en maltotriose. Hieruit kan geconcludeerd worden dat dit DP9-suiker een α(1→4) maltohexaose eenheid is die α(1→6) gesubstitueerd is met een maltotriose eenheid op de O-6 positie van drie verschillende α(1→4) gelinkte glucose residu's. Referenties (1) Van Landschoot, A., Vanbeneden, N., Machtelinckx, M., Stals, I. & Claeyssens, M. (2005) Pecularities of seven refermented Belgian strong ales and their corresponding industrial yeast. Cerevisia, 30, 3, 181-188. (2) Laroy W., Contreras R. and Callewaert N. (2006) Glycome mapping on DNA sequencing equipment. Nature Protocols 397-405. (3) Redmond J.W. and Packer N.H. (1999) The use of solid-phase extraction with graphitised carbon for the fractionation and purification of sugars. Carbohydrate Research 319, 74-79

VaLiCel: valorisation of (ligno-)cellulose residues and energy crops

Based on the estimated global annual production of biomass (1x1011 ton ≈ 2 ZJ ), theoretically plants could easily substitute for the annual 0.2 ZJ petroleum production. In only one decade, plant biomass could in the form of cellulose, hemicellulose, and lignin renew the 20 ZJ energy stored as conventional crude oil . Although the relatively high cost of both pretreatment and enzymes that catalyze cellulose hydrolysis [one-third of the cost of ethanol production from cellulose] still represents a major barrier to commercialization, cellulosic ethanol production is currently in the pilot plant stage, with more than 30 pilot plants being operated in both North-America and Europe. The process generally consists of a pretreatment step, a hydrolysis and a fermentation step. In these processes lignin is discharged as a byproduct that can be used to supply the process with energy. It is however generally believed that a successful bioenergy research policy should focus on biorefineries, co-producing transportation fuels, power, heat, added-value chemicals and materials from biomass. In the IWT TETRA [a grant program to stimulate innovation in Flemish companies through technology transfer] project VaLiCel, we have compared several pretreatment, hydrolysis and fermentation conditions using lignocellulosic residues (recycled paper pulp, loams & scutched short flax, wood (dust), grasses and agro-industrial waste) and dedicated energy crops (Miscanthus x giganteus, poplar and willow). The yield of fermentable sugars and ethanol is determined and compared. Ultimately, the aim is to know which lignocellulose waste is suited for complete saccharification to glucose and xylose and yields minor side products that are inhibitory to alcohol fermentation. From our results it is clear that by screening a collection of different biomass feedstocks, evaluating different pretreatment methods, screening a large set of yeast strains and optimizing the préculture and fermentation conditions, it is possible to produce second generation bio-ethanol in an economically viable way. First results of a case study will exemplify a possible integration strategy into existing industrial fuel producing plants. It is expected that the co-production of added-value products will lead to overall economic profitability in an integrated biorefinery process. In casu the valorisation of lignin -as a source for aromatic chemicals- can play a key role in the further development of lignocellulosic biomass conversion processes

Pilot scale recovery of lignin from black liquor and advanced characterization of the final product

Recently, the academic and industrial interest in lignin as a renewable resource for many valuable applications has been on the rise. However, the current biomass separation technologies are focused on obtaining high quality cellulose which can be further processed, e.g., in the paper industry, resulting in a lignin of rather low quality. Moreover, lignin recovery from black liquor is often accompanied with filter clogging and a severe flux decline, limiting the cost-efficiency of its valorization. In this work, the pilot scale recovery of lignin from a black liquor derived from a mild soda pulping process of Miscanthus x giganteus chips is studied with the aim to develop a straightforward procedure that yields a high quality final product. A first pilot scale experiment demonstrated the pH to be crucial for optimal precipitation. Moreover, adding an enzyme mixture containing cellulases, hemicellulases and beta-glucosidases, clearly enhanced the flocculation and filterability. Thorough characterization of the obtained lignin showed a native-like structure which can be related to the mild pulping conditions and revealed that the p-coumarates and ferulates were converted to the free acids as a result of the base catalyzed hydrolysis as well as the enzymatic cleavage of the ester linkages leading to the complete removal of the hydrophilic (poly)saccharides. Moreover, this resulted in a slightly more hydrophobic lignin material that was more amenable to flocculation. Building on lab scale experiments aimed at optimization of the process conditions, a second pilot scale experiment was performed resulting in improved precipitation and flocculation by means of acidification, an enzymatic treatment as well as the addition of a flocculant. This allowed for smooth filtration and resulted in a high purity of the isolated lignin