728 research outputs found
Genome-wide screening of haploid stem cells for the discovery of essential genes for marburg virus infection
No full textIdentification of host proteins that serve as viral entry factors has enabled insights into virus particle internalization, tropism, and pathogenesis. In this context, haploid cells allow the study of gene knockouts, since recessive mutations will show a clear phenotype due to the absence of a 2nd gene copy. The study of highly pathogenic viruses (HiPVs) present significant challenges due to the requirement for BSL-4 containments. Indeed it has been proposed to use the Vesicular Stomatitis Virus (VSV) pseudotyped with glycoprotein of HiPVs to study the viral entry pathway.
To identify host factors required for Marburg virus infection, we used a mammalian haploid embryonic stem cells library for infection with a recombinant VSV expressing the Marburg virus glycoprotein (rVSV-M) instead of the native glycoprotein.
While wild-type cells massively died upon rVSV-M infection, a high number of cells survived within the mutagenized cells. Surviving cells were analyzed by next generation sequencing to identify the mutated genes. Statistical analysis provided a list of 7 genes, potentially involved in rVSV-M entry, which will be validated by infection of specific KO clones. Their role on the viral life cycle will be investigated
High Precision Quantitative Proteomics Using iTRAQ on an LTQ Orbitrap: A New Mass Spectrometric Method Combining the Benefits of All
No full textThe development of quantitative techniques in mass spectrometry has generated the ability to systematically monitor protein expression. Isobaric tags for relative and absolute quantification (iTRAQ) have become a widely used tool for the quantification of proteins. However, application of iTRAQ methodology using ion traps and hybrid mass spectrometers containing an ion trap such as the LTQ-Orbitrap was not possible until the development of pulsed Q dissociation (PQD) and higher energy C-trap dissociation (HCD). Both methods allow iTRAQ-based quantification on an LTQ-Orbitrap but are less suited for protein identification at a proteomic scale than the commonly used collisional induced dissociation (CID) fragmentation. We developed an analytical strategy combining the advantages of CID and HCD, allowing sensitive and accurate protein identification and quantitation at the same time. In a direct comparison, the novel method outperformed PQD and HCD regarding its limit of detection, the number of identified peptides and the analytical precision of quantitation. The new method was applied to study changes in protein expression in mouse hearts upon transverse aortic constriction, a model for cardiac stress
Illustrierte Kriegsbeilage Nr. 50 1916 (Nr. 50 1916)
No full textILLUSTRIERTE KRIEGSBEILAGE NR. 50 1916
Illustrierte Kriegsbeilage (-)
Illustrierte Kriegsbeilage Nr. 50 1916 (Nr. 50 1916) ( - )
Für den Herrgott. ( - )
[2 Abb]: (1)General Falkenhayn. (2)Bilder aus Südtirol: Motiv aus Arco. ( - )
Stille Helden. (Militärseelsorge.) VII. ( - )
[Abb]: Agram vom Luftballon aus. ( - )
Die Welt der Surrogate. ( - )
[2 Abb]: (1)Der Krieg in der Geschichte: Belagerung von Kufstein. (2)Schwedisches Militär auf Ski. ( - )
[Abb]: Ministerpräsident Dr. Koerber. ( - )
Die Spinne als Wetterprophet. ( - )
Ist der Mars wirklich bewohnt? ( - )
[Gedicht]: Die Glocken. ( - )
[Abb]: Opfer des Krieges aus Oberösterreich. Karl Froschauer (Naarn), Hans Pühringer (Fall b. Wilhering), Josef Huber (Dörfling, Kronstorf), Johann Leitner (Müllerberg, Wallern). ( - )
[Abb]: Opfer des Krieges aus Oberösterreich. Friedrich Pokorny, Georg Mendl (Urfahr), Josef Hochholzer (Urfahr), Johann Wegscheider, Karl Niederholzer, Anna Just, Franz Weichselbaumer (Gaisberg, St. Martin i. M.), Michael Haselmeier (Plöcking, St. Martin i. M.), Franz Krammer (Hofstetten, St. Martin i. M.), Ferdinand Kießl (St. Martin i. M.), Josef Schwarz (Oberaigen, Hellmonsödt), Konrad Pilsl (Davidschlag, Hellmonsödt), Johann Fuchshuber (St. Florian), Johann Mühlbachler (Eibenberg, Liebenau), Ignaz Lindner (Mitterkirchen), Josef Jungwirth (Mollmannsreith, Kollerschlag i. Mühlkr.), Johann Weyrer (Lasberg), Michael Wögerer (Grub, Lasberg), Karl Dauerböck, Josef Preßlmair, Johann Aichhorn, Karl Pachinger, Karl Schütz (Walding), Martin Wiesinger (Rodl, Walding), Franz Weichselberger (St. Marienkirchen a. H.), Peter Adlmanseder (Koblstadt, St. Martin), Josef Penninger (Fritzging, Mehrnbach), Johann Neuhofer (Abstetten, Mehrnbach), Engelbert Gattermaier (Edt, Kirchheim), Ferdinand Lehner (Weg, Hofkirchen a. d. Tr.). ( -
Neuropathological assessment of the olfactory bulb and tract in individuals with COVID-19
No full textThe majority of patients with Parkinson disease (PD) experience a loss in their sense of smell and accumulate insoluble α-synuclein aggregates in their olfactory bulbs (OB). Subjects affected by a SARS-CoV-2-linked illness (COVID-19) also frequently experience hyposmia. We previously postulated that microglial activation as well as α-synuclein and tau misprocessing can occur during host responses following microbial encounters. Using semiquantitative measurements of immunohistochemical signals, we examined OB and olfactory tract specimens collected serially at autopsies between 2020 and 2023. Deceased subjects comprised 50 adults, which included COVID19 + patients (n = 22), individuals with Lewy body disease (e.g., PD; dementia with Lewy bodies (n = 6)), Alzheimer disease (AD; n = 3), and other neurodegenerative disorders (e.g., progressive supranuclear palsy (n = 2); multisystem atrophy (n = 1)). Further, we included neurologically healthy controls (n = 9), and added subjects with an inflammation-rich brain disorder as neurological controls (NCO; n = 7). When probing for microglial and histiocytic reactivity in the anterior olfactory nuclei (AON) by anti-CD68 immunostaining, scores were consistently elevated in NCO and AD cases. In contrast, microglial signals on average were not significantly altered in COVID19 + patients relative to healthy controls, although anti-CD68 reactivity in their OB and tracts declined with progression in age. Mild-to-moderate increases in phospho-α-synuclein and phospho-tau signals were detected in the AON of tauopathy- and synucleinopathy-afflicted brains, respectively, consistent with mixed pathology, as described by others. Lastly, when both sides were available for comparison in our case series, we saw no asymmetry in the degree of pathology of the left versus right OB and tracts. We concluded from our autopsy series that after a fatal course of COVID-19, microscopic changes in the rostral, intracranial portion of the olfactory circuitry -when present- reflected neurodegenerative processes seen elsewhere in the brain. In general, microglial reactivity correlated best with the degree of Alzheimer’s-linked tauopathy and declined with progression of age in COVID19 + patients
Genome wide functional genetics in haploid cells
No full textAbstractSome organisms such as yeast or males of social insects are haploid, i.e. they carry a single set of chromosomes, while haploidy in mammals is exclusively restricted to mature germ cells. A single copy of the genome provides the basis for genetic analyses where any recessive mutation of essential genes will show a clear phenotype due to the absence of a second gene copy. Most prominently, haploidy in yeast has been utilized for recessive genetic screens that have markedly contributed to our understanding of development, basic physiology, and disease. Somatic mammalian cells carry two copies of chromosomes (diploidy) that obscure genetic analysis. Near haploid human leukemic cells however have been developed as a high throughput screening tool. Although deemed impossible, we and others have generated mammalian haploid embryonic stem cells from parthenogenetic mouse embryos. Haploid stem cells open the possibility of combining the power of a haploid genome with pluripotency of embryonic stem cells to uncover fundamental biological processes in defined cell types at a genomic scale. Haploid genetics has thus become a powerful alternative to RNAi or CRISPR based screens
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