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5 result(s) for "Bokelmann, Lukas"
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Point-of-care bulk testing for SARS-CoV-2 by combining hybridization capture with improved colorimetric LAMP
Efforts to contain the spread of SARS-CoV-2 have spurred the need for reliable, rapid, and cost-effective diagnostic methods which can be applied to large numbers of people. However, current standard protocols for the detection of viral nucleic acids while sensitive, require a high level of automation and sophisticated laboratory equipment to achieve throughputs that allow whole communities to be tested on a regular basis. Here we present Cap-iLAMP (capture and improved loop-mediated isothermal amplification) which combines a hybridization capture-based RNA extraction of gargle lavage samples with an improved colorimetric RT-LAMP assay and smartphone-based color scoring. Cap-iLAMP is compatible with point-of-care testing and enables the detection of SARS-CoV-2 positive samples in less than one hour. In contrast to direct addition of the sample to improved LAMP (iLAMP), Cap-iLAMP prevents false positives and allows single positive samples to be detected in pools of 25 negative samples, reducing the reagent cost per test to ~1 Euro per individual. Current SARS-CoV-2 diagnostic methods are sensitive yet poorly suited to testing whole communities on a regular basis. Here the authors present Cap-iLAMP that tests gargle lavage samples with an improved colorimetric RT-LAMP.
A direct RT-qPCR approach to test large numbers of individuals for SARS-CoV-2
SARS-CoV-2 causes substantial morbidity and mortality in elderly and immunocompromised individuals, particularly in retirement homes, where transmission from asymptomatic staff and visitors may introduce the infection. Here we present a cheap and fast screening method based on direct RT-qPCR to detect SARS-CoV-2 in single or pooled gargle lavages (“mouthwashes”). This method detects individuals with large viral loads (Ct≤29) and we use it to test all staff at a nursing home daily over a period of three weeks in order to reduce the risk that the infection penetrates the facility. This or similar approaches can be implemented to protect hospitals, nursing homes and other institutions in this and future viral epidemics.
The complete mitochondrial genome of the hymenopteran hunting robber fly Dasypogon diadema
The complete mitochondrial genome of Dasypogon diadema (Insecta: Diptera) was sequenced using the Illumina MiSeq` platform. Its mt-genome spans over 16,947 bp with a GC content of 26.6% containing 13 protein-coding genes, 22 tRNA genes, and 2 rRNA genes. The phylogenetic relationship of Dasypogon diadema and 11 other dipteran species was reconstructed and the phylogenetic position confirmed.
A genetic analysis of the Gibraltar Neanderthals
The Forbes’ Quarry and Devil’s Tower partial crania from Gibraltar are among the first Neanderthal remains ever found. Here, we show that small amounts of ancient DNA are preserved in the petrous bones of the 2 individuals despite unfavorable climatic conditions. However, the endogenous Neanderthal DNA is present among an overwhelming excess of recent human DNA. Using improved DNA library construction methods that enrich for DNA fragments carrying deaminated cytosine residues, we were able to sequence 70 and 0.4 megabase pairs (Mbp) nuclear DNA of the Forbes’ Quarry and Devil’s Tower specimens, respectively, as well as large parts of the mitochondrial genome of the Forbes’ Quarry individual. We confirm that the Forbes’ Quarry individual was a female and the Devil’s Tower individual a male. We also show that the Forbes’ Quarry individual is genetically more similar to the ∼120,000-y-old Neanderthals from Scladina Cave in Belgium (Scladina I-4A) and Hohlenstein-Stadel Cave in Germany, as well as to a ∼60,000- to 70,000-y-old Neanderthal from Russia (Mezmaiskaya 1), than to a ∼49,000-y-old Neanderthal from El Sidrón (El Sidrón 1253) in northern Spain and other younger Neanderthals from Europe and western Asia. This suggests that the Forbes’ Quarry fossil predates the latter Neanderthals. The preservation of archaic human DNA in the warm coastal climate of Gibraltar, close to the shores of Africa, raises hopes for the future recovery of archaic human DNA from regions in which climatic conditions are less than optimal for DNA preservation.
Effect of iterative reconstruction and temporal averaging on contour sharpness in dynamic myocardial CT perfusion: Sub-analysis of the prospective 4D CT perfusion pilot study
Myocardial computed tomography perfusion (CTP) allows the assessment of the functional relevance of coronary artery stenosis. This study investigates to what extent the contour sharpness of sequences acquired by dynamic myocardial CTP is influenced by the following noise reduction methods: temporal averaging and adaptive iterative dose reduction 3D (AIDR 3D). Dynamic myocardial CT perfusion was conducted in 29 patients at a dose level of 9.5±2.0 mSv and was reconstructed with both filtered back projection (FBP) and strong levels of AIDR 3D. Temporal averaging to reduce noise was performed as a post-processing step by combining two, three, four, six and eight original consecutive 3D datasets. We evaluated the contour sharpness at four distinct edges of the left-ventricular myocardium based on two different approaches: the distance between 25% and 75% of the maximal grey value (d) and the slope in the contour (m). Iterative reconstruction reduced contour sharpness: both measures of contour sharpness performed better for FBP than for AIDR 3D (d = 1.7±0.4 mm versus 2.0±0.5 mm, p>0.059 at all edges; m = 255.9±123.9 HU/mm versus 160.6±123.5 HU/mm; p<0.023 for all edges). Increasing levels of temporal averaging degraded contour sharpness. When FBP reconstruction was applied, contour sharpness was best without temporal averaging (d = 1.7±0.4 mm, m = 255.9±123.9 HU/mm) and poorest for the strongest levels of temporal averaging (d = 2.1±0.3 mm, m = 142.2±104.9 HU/mm; comparison between lowest and highest temporal averaging level: for d p>0.052 at all edges and for m p<0.001 at all edges). The use of both temporal averaging and iterative reconstruction degrades objective contour sharpness parameters of dynamic myocardial CTP. Thus, further advances in image processing are needed to optimise contour sharpness of 4D myocardial CTP.