Analyse der Aktivierung des angeborenen Immunsystems durch Candida spp. mittels Lebendzellmikroskopie

Candida albicans und Candida glabrata sind die beiden wichtigsten Erreger von invasiven Infektionen im Genus Candida. Obwohl beide Spezies derselben Gattung angehören, unterscheiden sich ihre Virulenzmechanismen erheblich. Ziel dieser Arbeit war es, mit Hilfe der Lebendzellmikroskopie zeitaufgelöste Aufnahmen zu generieren, die eine detaillierte Untersuchung der Aktivierung von humanen neutrophilen Granulozyten (PMN) durch die beiden Arten erlauben. Es konnte gezeigt werden, dass C. albicans eine signifikant stärkere PMN-Aktivierung induziert als C. glabrata. Dennoch genügte die moderate Aktivierung der PMN, um phagozytierte C. glabrata effektiver abzutöten als C. albicans. Lebendzellmikroskopisch konnte belegt werden, dass C. albicans signifikant häufiger phagozytiert wurde als C. glabrata. Dies war nicht auf den fehlenden Kontakt zwischen den Immunzellen und C. glabrata zurückzuführen, sondern auf eine geringere Phagozytose-Effektivität der PMN nach Kontakt mit dem Pilz. Neben der gesteigerten Phagozytose-Rate induzierte C. albicans auch eine höhere Motilität der PMN als C. glabrata. Dies ist unter anderem auf eine stärkere Sekretion von PMN Chemoattraktoren wie IL-8 und GROα zurückzuführen. Die Konfrontation von PMN mit C. glabrata führte dahingegen zur Sekretion eines Zytokinprofils, das auf eine mögliche Involvierung von Monozyten in der Abwehr von C. glabrata Infektionen hindeutet. Zusammenfassend hat diese Arbeit durch die Etablierung der Lebendzellmikroskopie zu einem besseren Verständnis der Interaktion zwischen PMN und den beiden Candida spp. beigetragen. Durch die Entwicklung einer vollautomatisierten PMN Trackingmethode wurde ein wichtiger Grundstein für die zukünftige Etablierung einer vollautomatisierten Bildanalyse der Pathogen-Wirt-Interaktion gelegt

Constraining the mass of dark photons and axion-like particles through black-hole superradiance

Ultralight bosons and axion-like particles appear naturally in different scenarios and could solve some long-standing puzzles. Their detection is challenging, and all direct methods hinge on unknown couplings to the Standard Model of particle physics. However, the universal coupling to gravity provides model-independent signatures for these fields. We explore here the superradiant instability of spinning black holes triggered in the presence of such fields. The instability taps angular momentum from and limits the maximum spin of astrophysical black holes. We compute, for the first time, the spectrum of the most unstable modes of a massive vector (Proca) field for generic black-hole spin and Proca mass. The observed stability of the inner disk of stellar-mass black holes can be used to derive \emph{direct} constraints on the mass of dark photons in the mass range $10^{-13}\,{\rm eV}\lesssim m_V \lesssim 3\times 10^{-12}\,{\rm eV}$. By including also higher azimuthal modes, similar constraints apply to axion-like particles in the mass range $6\times10^{-13}\,{\rm eV}\lesssim m_{\rm ALP} \lesssim 10^{-11}\, {\rm eV}$. Likewise, mass and spin distributions of supermassive BHs --~as measured through continuum fitting, K$\alpha$ iron line, or with the future space-based gravitational-wave detector LISA~-- imply indirect bounds in the mass range approximately $10^{-19}\,{\rm eV}\lesssim m_V, m_{\rm ALP} \lesssim 10^{-13}\, {\rm eV}$, for both axion-like particles and dark photons. Overall, superradiance allows to explore a region of approximately $8$ orders of magnitude in the mass of ultralight bosons

EMIP: The eye movements in programming dataset

A large dataset that contains the eye movements of N=216 programmers of different experience levels captured during two code comprehension tasks is presented. Data are grouped in terms of programming expertise (from none to high) and other demographic descriptors. Data were collected through an international collaborative effort that involved eleven research teams across eight countries on four continents. The same eye tracking apparatus and software was used for the data collection. The Eye Movements in Programming (EMIP) dataset is freely available for download. The varied metadata in the EMIP dataset provides fertile ground for the analysis of gaze behavior and may be used to make novel insights about code comprehension

High Mobility Group Box 1 Protein in Cerebral Thromboemboli

High-mobility group box 1 protein (HMGB1) is a damage-associated molecular pattern (DAMP) involved in neutrophil extracellular trap (NET) formation and thrombosis. NETs are regularly found in cerebral thromboemboli. We here analyzed associated HMGB1 expression in human thromboemboli retrieved via mechanical thrombectomy from 37 stroke patients with large vessel occlusion. HMGB1 was detected in all thromboemboli, accounting for 1.7% (IQR 0.6–6.2%) of the total thromboemboli area and was found to be colocalized with neutrophils and NETs and in spatial proximity to platelets. Correlation analysis revealed that the detection of HMGB1 was strongly related to the number of neutrophils (r = 0.58, p = 0.0002) and platelets (r = 0.51, p = 0.001). Our results demonstrate that HMGB1 is a substantial constituent of thromboemboli causing large vessel occlusion stroke

Immunohistological Analysis of Neutrophils and Neutrophil Extracellular Traps in Human Thrombemboli Causing Acute Ischemic Stroke

Ischemic stroke caused by thromboembolic occlusion of large cerebral arteries, such as the internal carotid (ICA) and/or the middle cerebral artery (MCA), is treated by mechanical thrombectomy (MT). MT allows salvage of the vessel-occluding thrombemboli, which most frequently originate from the left atrium or the left ventricle of the heart or from sites of plaque rupture within large arteries above the heart. Clot composition may influence the efficacy of (intravenous) thrombolysis and MT, respectively. We analyzed 37 human thrombemboli obtained from acute ischemic stroke patients during MT with special emphasis on histological staining of neutrophils and neutrophil extracellular traps (NETs). We found neutrophils as the main cellular component of cerebral thrombemboli but encountered considerable morphological heterogeneity. Neutrophils accumulated in the border region of fibrin-rich structures indicating possible interaction of neutrophils with distinct structural thrombembolus components. Web-like NETs were found in 35 of 37 thrombemboli in varying amounts. NETs were almost exclusively found within fibrin-rich areas. Importantly, stroke etiology, age and present oral anticoagulation was associated with morphological patterns and the amount of neutrophils. Correlation of histological data and imaging data revealed that relative Hounsfield units of cerebral thrombemboli positively correlated with the amount of red blood cells. In summary, our results demonstrate that neutrophils and NETs are substantial constituents of cerebral thrombemboli and contribute to their structural complexity

Safety and Effectiveness of the New Generation APERIO® Hybrid Stent-retriever Device in Large Vessel Occlusion Stroke

Background It is unknown whether technological advancement of stent-retriever devices influences typical observational indicators of safety or effectiveness. Methods Observational retrospective study of APERIO® (AP) vs. new generation APERIO® Hybrid (APH) (Acandis®, Pforzheim, Germany) stent-retriever device (01/2019–09/2020) for mechanical thrombectomy (MT) in large vessel occlusion (LVO) stroke. Primary effectiveness endpoint was successful recanalization eTICI (expanded Thrombolysis In Cerebral Ischemia) ≥ 2b67, primary safety endpoint was occurrence of hemorrhagic complications after MT. Secondary outcome measures were time from groin puncture to first pass and successful reperfusion, and the total number of passes needed to achieve the final recanalization result. Results A total of 298 patients with LVO stroke who were treated by MT matched the inclusion criteria: 148 patients (49.7%) treated with AP vs. 150 patients (50.3%) treated with new generation APH. Successful recanalization was not statistically different between both groups: 75.7% for AP vs. 79.3% for APH; p = 0.450. Postinterventional hemorrhagic complications and particularly subarachnoid hemorrhage as the entity possibly associated with stent-retriever device type was significantly less frequent in the group treated with the APH: 29.7% for AP and 16.0% for APH; p = 0.005; however, rates of symptomatic hemorrhage with clinical deterioration and in domo mortality were not statistically different. Neither the median number of stent-retriever passages needed to achieve final recanalization, time from groin puncture to first pass, time from groin puncture to final recanalization nor the number of cases in which successful recanalization could only be achieved by using a different stent-retriever as bail-out device differed between both groups. Conclusion In the specific example of the APERIO® stent-retriever device, we observed that further technological developments of the new generation device were not associated with disadvantages with respect to typical observational indicators of safety or effectiveness

High mobility group box 1 protein in cerebral thromboemboli

High-mobility group box 1 protein (HMGB1) is a damage-associated molecular pattern (DAMP) involved in neutrophil extracellular trap (NET) formation and thrombosis. NETs are regularly found in cerebral thromboemboli. We here analyzed associated HMGB1 expression in human thromboemboli retrieved via mechanical thrombectomy from 37 stroke patients with large vessel occlusion. HMGB1 was detected in all thromboemboli, accounting for 1.7% (IQR 0.6–6.2%) of the total thromboemboli area and was found to be colocalized with neutrophils and NETs and in spatial proximity to platelets. Correlation analysis revealed that the detection of HMGB1 was strongly related to the number of neutrophils (r = 0.58, p = 0.0002) and platelets (r = 0.51, p = 0.001). Our results demonstrate that HMGB1 is a substantial constituent of thromboemboli causing large vessel occlusion stroke

Immune cells invade the collateral circulation during human stroke: prospective replication and extension

It remains unclear if principal components of the local cerebral stroke immune response can be reliably and reproducibly observed in patients with acute large-vessel-occlusion (LVO) stroke. We prospectively studied a large independent cohort of n = 318 consecutive LVO stroke patients undergoing mechanical thrombectomy during which cerebral blood samples from within the occluded anterior circulation and systemic control samples from the ipsilateral cervical internal carotid artery were obtained. An extensive protocol was applied to homogenize the patient cohort and to standardize the procedural steps of endovascular sample collection, sample processing, and laboratory analyses. N = 58 patients met all inclusion criteria. (1) Mean total leukocyte counts were significantly higher within the occluded ischemic cerebral vasculature (I) vs. intraindividual systemic controls (S): +9.6%, I: 8114/µL ± 529 vs. S: 7406/µL ± 468, p = 0.0125. (2) This increase was driven by neutrophils: +12.1%, I: 7197/µL ± 510 vs. S: 6420/µL ± 438, p = 0.0022. Leukocyte influx was associated with (3) reduced retrograde collateral flow (R$^2$ = 0.09696, p = 0.0373) and (4) greater infarct extent (R$^2$ = 0.08382, p = 0.032). Despite LVO, leukocytes invade the occluded territory via retrograde collateral pathways early during ischemia, likely compromising cerebral hemodynamics and tissue integrity. This inflammatory response can be reliably observed in human stroke by harvesting immune cells from the occluded cerebral vascular compartment

Immune Cells Invade the Collateral Circulation during Human Stroke: Prospective Replication and Extension

It remains unclear if principal components of the local cerebral stroke immune response can be reliably and reproducibly observed in patients with acute large-vessel-occlusion (LVO) stroke. We prospectively studied a large independent cohort of n = 318 consecutive LVO stroke patients undergoing mechanical thrombectomy during which cerebral blood samples from within the occluded anterior circulation and systemic control samples from the ipsilateral cervical internal carotid artery were obtained. An extensive protocol was applied to homogenize the patient cohort and to standardize the procedural steps of endovascular sample collection, sample processing, and laboratory analyses. N = 58 patients met all inclusion criteria. (1) Mean total leukocyte counts were significantly higher within the occluded ischemic cerebral vasculature (I) vs. intraindividual systemic controls (S): +9.6%, I: 8114/µL ± 529 vs. S: 7406/µL ± 468, p = 0.0125. (2) This increase was driven by neutrophils: +12.1%, I: 7197/µL ± 510 vs. S: 6420/µL ± 438, p = 0.0022. Leukocyte influx was associated with (3) reduced retrograde collateral flow (R2 = 0.09696, p = 0.0373) and (4) greater infarct extent (R2 = 0.08382, p = 0.032). Despite LVO, leukocytes invade the occluded territory via retrograde collateral pathways early during ischemia, likely compromising cerebral hemodynamics and tissue integrity. This inflammatory response can be reliably observed in human stroke by harvesting immune cells from the occluded cerebral vascular compartment