State-of-the-art analytical methods of viral infections in human lung organoids

Human-based organ models can provide strong predictive value to investigate the tropism, virulence, and replication kinetics of viral pathogens. Currently, such models have received widespread attention in the study of SARS-CoV-2 causing the COVID-19 pandemic. Applicable to a large set of organoid models and viruses, we provide a step-by-step work instruction for the infection of human alveolar-like organoids with SARS-CoV-2 in this protocol collection. We also prepared a detailed description on state-of-the-art methodologies to assess the infection impact and the analysis of relevant host factors in organoids. This protocol collection consists of five different sets of protocols. Set 1 describes the protein extraction from human alveolar-like organoids and the determination of protein expression of angiotensin-converting enzyme 2 (ACE2), transmembrane serine protease 2 (TMPRSS2) and FURIN as exemplary host factors of SARS-CoV-2. Set 2 provides detailed guidance on the extraction of RNA from human alveolar-like organoids and the subsequent qPCR to quantify the expression level of ACE2, TMPRSS2, and FURIN as host factors of SARS-CoV-2 on the mRNA level. Protocol set 3 contains an in-depth explanation on how to infect human alveolar-like organoids with SARS-CoV-2 and how to quantify the viral replication by plaque assay and viral E gene-based RT-qPCR. Set 4 provides a step-by-step protocol for the isolation of single cells from infected human alveolar-like organoids for further processing in single-cell RNA sequencing or flow cytometry. Set 5 presents a detailed protocol on how to perform the fixation of human alveolar-like organoids and guides through all steps of immunohistochemistry and in situ hybridization to visualize SARS-CoV-2 and its host factors. The infection and all subsequent analytical methods have been successfully validated by biological replications with human alveolar-like organoids based on material from different donors

Inhibition of the Soluble Epoxide Hydrolase Promotes Albuminuria in Mice with Progressive Renal Disease

Epoxyeicotrienoic acids (EETs) are cytochrome P450-dependent anti-hypertensive and anti-inflammatory derivatives of arachidonic acid, which are highly abundant in the kidney and considered reno-protective. EETs are degraded by the enzyme soluble epoxide hydrolase (sEH) and sEH inhibitors are considered treatment for chronic renal failure (CRF). We determined whether sEH inhibition attenuates the progression of CRF in the 5/6-nephrectomy model (5/6-Nx) in mice. 5/6-Nx mice were treated with a placebo, an ACE-inhibitor (Ramipril, 40 mg/kg), the sEH-inhibitor cAUCB or the CYP-inhibitor fenbendazole for 8 weeks. 5/6-Nx induced hypertension, albuminuria, glomerulosclerosis and tubulo-interstitial damage and these effects were attenuated by Ramipril. In contrast, cAUCB failed to lower the blood pressure and albuminuria was more severe as compared to placebo. Plasma EET-levels were doubled in 5/6 Nx-mice as compared to sham mice receiving placebo. Renal sEH expression was attenuated in 5/6-Nx mice but cAUCB in these animals still further increased the EET-level. cAUCB also increased 5-HETE and 15-HETE, which derive from peroxidation or lipoxygenases. Similar to cAUCB, CYP450 inhibition increased HETEs and promoted albuminuria. Thus, sEH-inhibition failed to elicit protective effects in the 5/6-Nx model and showed a tendency to aggravate the disease. These effects might be consequence of a shift of arachidonic acid metabolism into the lipoxygenase pathway

Nothing Lasts Forever: Environmental Discourses on the Collapse of Past Societies

The study of the collapse of past societies raises many questions for the theory and practice of archaeology. Interest in collapse extends as well into the natural sciences and environmental and sustainability policy. Despite a range of approaches to collapse, the predominant paradigm is environmental collapse, which I argue obscures recognition of the dynamic role of social processes that lie at the heart of human communities. These environmental discourses, together with confusion over terminology and the concepts of collapse, have created widespread aporia about collapse and resulted in the creation of mixed messages about complex historical and social processes

Sensitivity of the Freie Universität Berlin Climate Middle Atmosphere Model (FUB-CMAM) to different gravity-wave drag parameterisations

International audienceWe report the sensitivity of the Berlin Climate Middle Atmosphere Model (CMAM) to different gravity-wave (GW) parameterisations. We perform five perpetual January experiments: 1) Rayleigh friction (RF) (control), 2) non-orographic GWs, 3) orographic GWs, 4) orographic and non-orographic GWs with no background stress, and 5) as for 4) but with background stress. We also repeat experiment 4) but for July conditions. Our main aim is to improve the model climatology by introducing orographic and non-orographic parameterisations and to investigate the individual effect of these schemes in the Berlin CMAM. We compare with an RF control to determine the improvement upon a previously-published model version employing RF. Results are broadly similar to previously-published works. The runs having both orographic and non-orographic GWs produce a statistically-significant warming of 4-8K in the wintertime polar lower stratosphere. These runs also feature a cooling of the warm summer pole in the mesosphere by 10-15K, more in line with observations. This is associated with the non-orographic GW scheme. This scheme is also associated with a heating feature in the winter polar upper stratosphere directly below the peak GW-breaking region. The runs with both orographic and non-orographic GWs feature a statistically-significant deceleration in the polar night jet (PNJ) of 10-20ms-1 in the lower stratosphere. Both orographic and non-orographic GWs individually produce some latitudinal tilting of the polar jet with height, although the main effect comes from the non-orographic waves. The resulting degree of tilt, although improved, is nevertheless still weaker than that observed. Accordingly, wintertime variability in the zonal mean wind, which peaks at the edge of the vortex, tends to maximise too far polewards in the model compared with observations. Gravity-planetary wave interaction leads to a decrease in the amplitudes of stationary planetary waves 1 and 2 by up to 50% in the upper stratosphere and mesosphere, more in line with observations. Comparing modelled and observed Eliassen-Palm fluxes suggests that planetary wave (PW) breaking occurs too far polewards in the model. The wind and temperature changes are consistent with changes in the Brewer-Dobson (BD) circulation. Results suggest that the effect of enforcing a minimum background wave stress in the McFarlane scheme could be potentially important. In the Southern Hemisphere (SH) in July, the GW schemes had only a small impact on the high-latitude lower stratosphere but there featured strong warming near 0.1hPa

Health related quality of life of former very preterm infants in adulthood

Aim:  To assess health related quality of life of young adults born very preterm compared with a term control group. Methods:  A cohort of preterm infants <1250 g and a term control group, both born between 1983-1985, were surveyed as adults at the median age of 23 years. Questionnaires including the SF 36 and a modified lifestyle questionnaire assessed quality of life, health attitudes, height and weight, chronic diseases, medication and drug consumption. Results:  52 preterms and 75 controls matched for age and sex participated in the study. There were no significant differences in the quality of life as assessed by SF 36. Former preterms were significantly smaller than their term controls but not so for body mass index. The overall consumption of illicit drugs was significantly lower in former preterms. Moreover, former preterms went significantly less often in for sports. There was a trend for higher prevalence of chronic diseases in male compared to female preterms, but their use of medication was significantly lower. Conclusion:  Adults, born very preterm show no significant differences in their quality of life when compared to controls in early adulthood. However, based on their lifestyle and health disadvantages, male preterm subjects constitute a risk group when entering early adulthood with a clear need for continued attention

Sensitivity of the Freie Universität Berlin Climate Middle Atmosphere Model (FUB-CMAM) to different gravity-wave drag parameterisations

We report the sensitivity of the Berlin Climate Middle Atmosphere Model (CMAM) to different gravity-wave (GW) parameterisations. We perform five perpetual January experiments: 1) Rayleigh friction (RF) (control), 2) non-orographic GWs, 3) orographic GWs, 4) orographic and non-orographic GWs with no background stress, and 5) as for 4) but with background stress. We also repeat experiment 4) but for July conditions. Our main aim is to improve the model climatology by introducing orographic and non-orographic parameterisations and to investigate the individual effect of these schemes in the Berlin CMAM. We compare with an RF control to determine the improvement upon a previously-published model version employing RF. Results are broadly similar to previously-published works. The runs having both orographic and non-orographic GWs produce a statistically-significant warming of 4-8K in the wintertime polar lower stratosphere. These runs also feature a cooling of the warm summer pole in the mesosphere by 10-15K, more in line with observations. This is associated with the non-orographic GW scheme. This scheme is also associated with a heating feature in the winter polar upper stratosphere directly below the peak GW-breaking region. The runs with both orographic and non-orographic GWs feature a statistically-significant deceleration in the polar night jet (PNJ) of 10-20ms-1 in the lower stratosphere. Both orographic and non-orographic GWs individually produce some latitudinal tilting of the polar jet with height, although the main effect comes from the non-orographic waves. The resulting degree of tilt, although improved, is nevertheless still weaker than that observed. Accordingly, wintertime variability in the zonal mean wind, which peaks at the edge of the vortex, tends to maximise too far polewards in the model compared with observations. Gravity-planetary wave interaction leads to a decrease in the amplitudes of stationary planetary waves 1 and 2 by up to 50% in the upper stratosphere and mesosphere, more in line with observations. Comparing modelled and observed Eliassen-Palm fluxes suggests that planetary wave (PW) breaking occurs too far polewards in the model. The wind and temperature changes are consistent with changes in the Brewer-Dobson (BD) circulation. Results suggest that the effect of enforcing a minimum background wave stress in the McFarlane scheme could be potentially important. In the Southern Hemisphere (SH) in July, the GW schemes had only a small impact on the high-latitude lower stratosphere but there featured strong warming near 0.1hPa

Untersuchungen zum Erscheinungsbild von Materialfehlern in holografischen Interferogrammen

In holographic nondestructive testing, material faults are recognised by the identification of characteristic fringe irregularities. These partial patterns are caused by anomalous deformation of the object surface. Experiments and numerical simulations led to the hypothesis that the appearance of material faults in interference patterns can be reduced to a finite set of typical fringe irregularities. In the present paper the proof of this is given. A mathematical model of the holographic-interferometric image generation process is used. Interference fringes are considered as contour lines of interference phase which carries information about the deformation of the object surface. A relationship between contour lines and plane, autonomous systems of ordinary differential equations is found. Fringes correspond to solutions of initial value problems where the differential equations are defined by the interference phase and the initial value is determined by any point on the line and the fringe order. Because of their special structure the derived systems of differential equations are Hamiltonian systems. Together with knowledge about the mechanical behaviour of technical objects exposed to small loads which are used in nondestructive testing the chosen approach allows the above mentioned prove. The classification of stationary points of the interference phase #delta# (i.e. grad#delta#=0) corresponds to the consideration of critical points of the derived Hamiltonian system. With the addition of properties of regular points where grad#delta##not =#0 defines a fringe direction a list of all partial patterns for objects with macroscopically smooth surface results. Investigations on violations of the macroscopic smoothness complete the proof of the finite set of fringe irregularities. (orig.)Available from TIB Hannover: RR 9103(11) / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekSIGLEDEGerman