Takotsubo syndrome before and during the COVID-19 pandemic in Austria: a retrospective cohort study (TOSCA-19).

AIMS: This study aims to investigate the prevalence of Takotsubo syndrome (TTS) as a percentage of the total number of acute coronary syndrome (ACS), including non-STE-elevation myocardial infarction and ST-elevation myocardial infarction, as well as the short-term outcome of TTS patients before and during the COVID-19 pandemic. METHODS AND RESULTS: We compared patients from two different periods: (i) Period 1 (before the COVID-19 pandemic): 1 March to 30 December 2019, and (ii) Period 2 (during the COVID-19 pandemic): 1 March to 30 December 2020. The retrospective database was created from the archives of the participating hospitals or electronic hospital systems by trained medical personnel. The subjects' medical history, cardiovascular risk factors, laboratory values, echocardiography findings, and an in-hospital outcome were variables of interest. Furthermore, propensity score matching analysis was performed to evaluate the short-term prognosis in TTS and ACS patients. Altogether six Austrian centres-(i) 3rd Medical Department of Cardiology and Intensive Care Medicine, Clinic Ottakring, Vienna, Austria; (ii) 5th Medical Department of Cardiology, Clinic Favoriten, Vienna, Austria; (iii) 2nd Medical Department, Hanusch Hospital, Vienna, Austria; (iv) University Clinic of Internal Medicine III, Cardiology and Angiology, Medical University of Innsbruck, Austria; (v) Department of Cardiology, University Hospital Graz, Graz, Austria; (vi) Department of Cardiology and Intensive Medicine, Kepler University Clinic, Linz, Austria-participated in the study. During period 1, 87 (3.5%) patients out of 2482 ACS patients had TTS in all participating centres. During period 2, 71 (2.7%) patients out of 2572 ACS patients had TTS in all participating centres. Accordingly, the prevalence of TTS remained stable irrespective of potential psychologic stress during the COVID pandemic. Furthermore, the baseline characteristics of TTS patients did not change during the COVID-19 pandemic. The prevalence of in-hospital complications [cardiogenic shock (4.6% vs. 4.3%, P = 0.925), ventricle thrombus (1.1% vs. 1.4%, P = 0.885) and in-hospital bleeding (3.4% vs. 1.4%, P = 0.417)] remained stable. The all-cause in-hospital mortality of TTS patients did not change during the COVID-19 pandemic [χ2 (2) = 0.058, P = 0.810]. Moreover, a propensity score matching analysis of all-cause in-hospital mortality between matched TTS and ACS patients showed higher in-hospital mortality in ACS patients during COVID-19 pandemic (P = 0.043). CONCLUSIONS: Despite the well-known increased psychologic stress during the COVID-19 pandemic, the prevalence of TTS during the COVID-19 pandemic and the short-term clinical outcome in Austria remained unimpacted.

Neutrophile-Lymphocyte Ratio and Outcome in Takotsubo Syndrome.

Background: Takotsubo syndrome (TTS) is an important type of acute heart failure with significant risk of acute complications and death. In this analysis we sought to identify predictors for in-hospital clinical outcome in TTS patients and present long-term outcomes. Methods: In this analysis from the Austrian national TTS registry, univariable and multivariable analyses were performed to identify significant predictors for severe in-hospital complications requiring immediate invasive treatment or leading to irreversible damage, such as cardiogenic shock, intubation, stroke, arrhythmias and death. Furthermore, the influence of independent predictors on long-term survival was evaluated. Results: A total of 338 patients (median age 72 years, 86.9% female) from six centers were included. Severe in-hospital complications occurred in 14.5% of patients. In multivariable analysis, high neutrophile-lymphocyte-ratio (NLR; OR 1.04 [95% CI 1.02−1.07], p = 0.009) and low LVEF (OR 0.92 [0.90−0.95] per %, p < 0.001) were significant predictors of severe in-hospital complications. Both the highest NLR tercile and the lowest LVEF tercile were significantly associated with reduced 5-year survival. Conclusions: Low LVEF and high NLR at admission were independently associated with increased in-hospital complications and reduced long-term survival in TTS patients. NLR is a new easy-to-measure tool to predict worse short- and long-term outcome after TTS.

A novel Tetrahymena thermophila sterol C-22 desaturase belongs to the fatty acid hydroxylase/desaturase superfamily.

Sterols in eukaryotic cells play important roles in modulating membrane fluidity and in cell signaling and trafficking. During evolution, a combination of gene losses and acquisitions gave rise to an extraordinary diversity of sterols in different organisms. The sterol C-22 desaturase identified in plants and fungi as a cytochrome P-450 monooxygenase evolved from the first eukaryotic cytochrome P450 and was lost in many lineages. Although the ciliate Tetrahymena thermophila desaturates sterols at the C-22 position, no cytochrome P-450 orthologs are present in the genome. Here, we aim to identify the genes responsible for the desaturation as well as their probable origin. We used gene knockout and yeast heterologous expression approaches to identify two putative genes, retrieved from a previous transcriptomic analysis, as sterol C-22 desaturases. Furthermore, we demonstrate using bioinformatics and evolutionary analyses that both genes encode a novel type of sterol C-22 desaturase that belongs to the large fatty acid hydroxylase/desaturase superfamily and the genes originated by genetic duplication prior to functional diversification. These results stress the widespread existence of nonhomologous isofunctional enzymes among different lineages of the tree of life as well as the suitability for the use of T. thermophila as a valuable model to investigate the evolutionary process of large enzyme families.

Reactive Nitrogen Hotspots Related to Microscale Heterogeneity in Biological Soil Crusts.

Biocrusts covering drylands account for major fractions of terrestrial biological nitrogen fixation and release large amounts of gaseous reactive nitrogen (Nr) as nitrous acid (HONO) and nitric oxide (NO). Recent investigations suggested that aerobic and anaerobic microbial nitrogen transformations occur simultaneously upon desiccation of biocrusts, but the spatio-temporal distribution of seemingly contradictory processes remained unclear. Here, we explore small-scale gradients in chemical concentrations related to structural characteristics and organism distribution. X-ray microtomography and fluorescence microscopy revealed mixed pore size structures, where photoautotrophs and cyanobacterial polysaccharides clustered irregularly in the uppermost millimeter. Microsensor measurements showed strong gradients of pH, oxygen, and nitrite, nitrate, and ammonium ion concentrations at micrometer scales in both vertical and lateral directions. Initial oxygen saturation was mostly low (∼30%) at full water holding capacity, suggesting widely anoxic conditions, and increased rapidly upon desiccation. Nitrite concentrations (∼6 to 800 µM) and pH values (∼6.5 to 9.5) were highest around 70% WHC. During further desiccation they decreased, while emissions of HONO and NO increased, reaching maximum values around 20% WHC. Our results illustrate simultaneous, spatially separated aerobic and anaerobic nitrogen transformations, which are critical for Nr emissions, but might be impacted by future global change and land management.

Long-term outcome in patients with takotsubo syndrome : A single center study from Vienna.

BACKGROUND: There is an increasing amount of evidence suggesting multiple fatal complications in takotsubo syndrome; however, findings on the long-term outcome are scarce and show inconsistent evidence. METHODS: This is a single center study of long-term prognosis in takotsubo patients admitted to the Klinik Ottakring, Vienna, Austria, from September 2006 to August 2019. We investigated the clinical features, prognostic factors and outcome of patients with takotsubo syndrome. Furthermore, survivors and non-survivors and patients with a different cause of death were compared. RESULTS: Overall, 147 patients were included in the study and 49 takotsubo patients (33.3%) died during the follow-up, with a median of 126 months. The most common cause of death was a non-cardiac cause (71.4% of all deaths), especially malignancies (26.5% of all deaths). Moreover, non-survivors were older and more often men with more comorbidities (chronic kidney disease, malignancy). Patients who died because of cardiovascular disease were older and more often women than patients who died due to non-cardiovascular cause. Adjusted analysis showed no feature of an independent predictor of cardiovascular mortality for takotsubo patients. Female gender (HR = 0.32, CI: 0.16-0.64, p < 0.001), cancer (HR = 2.35, CI: 1.15-4.8, p = 0.019) and chronic kidney disease (HR = 2.61, CI: 1.11-6.14, p = 0.028) showed to be independent predictors of non-cardiovascular mortality. CONCLUSION: Long-term prognosis of takotsubo patients is not favorable, mainly due to noncardiac comorbidities. Hence, consequent outpatient care in regular intervals after a takotsubo event based on risk factor control and early detection of malignancies seems justified.

3D imaging of bean seeds: Correlations between hilum region structures and hydration kinetics.

X-ray micro-CT imaging has been applied successfully in food science and seed research due to its capacity to provide very small details of seed traits that are very complex to score. The micropyle and the tissues of the hilum region of bean seeds are recognized as structures which play an important role in hydration process. This latter influences, in turn, not only germination but also the cooking and industrial processing. Nevertheless, the role of each structure of the bean seeds is yet to be fully understood. Moreover such traits are never been quantified by using 3D imaging approaches. In this work, seeds of four ancient Italian landraces of beans have been scanned by X-ray microCT with a twofold approach: bulk scans for whole seed imaging and single seed scans for internal traits measurements. Then water uptake tests have been performed. The different structures composing the hilum region of the beans have been imaged and characterized. The two-dimensional and the three-dimensional morphometric traits have been correlated with parameters of hydration models by Principal Component Analysis (PCA) and Pearson coefficients. Micropyle groove area was the trait most influencing the very initial hydration rates while the hilum groove area was the best correlated with the overall infiltration behavior. The internal free space was the trait best correlated with the moisture at equilibrium. Moreover, strophiole shape resulted the most suitable internal trait for univocal identification of the four landraces. Overall results give a contribution to the understanding of the role of hilum region structures in bean seeds hydration process and show novel morphological traits useful for identification of local bean landraces.

Automatic cell identification and counting of leaf epidermis for plant phenotyping.

The counting of leaf epidermal cells is useful to study the plant developmental changes produced by environmental or genetic factors. The scanning electron microscopy can be used, but it is expensive and time-consuming. Methods using optical microscopy are also available, but they still require leaves pre-treatment and manual cell identification. We propose a quick and simple method for counting leaf epidermal cells without leaf treatments and based on automated cell identification and marking. It allows to highly improve the representativeness of leaf epidermis screening, aiming at a high-throughput plant phenotyping approach.•The leaves are pressed between two glass slides without any pre-treatment and digital micrographs are acquired under incident light.•Epidermal cells are automatically identified and counted by means of a "macro" of ImageJ•The cell count obtained applying the procedure of image processing is very close to that obtainable by manual cell identification.

Chemotropic vs Hydrotropic Stimuli for Root Growth Orientation in Microgravity.

Understanding how plants respond to spaceflight and extraterrestrial environments is crucial to develop life-support systems intended for long-term human explorations. Gravity is a main factor influencing root development and orientation, typically masking other tropisms. Considering that reduced levels of gravity affect many plant responses in space, the interaction of other tropic stimuli in microgravity represents the frontier to be investigated aiming at life-support systems optimization. In this paper we report on MULTITROP (Multiple-Tropism: interaction of gravity, nutrient and water stimuli for root orientation in microgravity), an experiment performed on the International Space Station during the Expedition 52/53. Scientific aim of the experiment was to disentangle hydrotropism from chemotropism for root orientation in absence of the gravity stimulus. Among several species relevant to space farming, Daucus carota was selected for the experiment because of its suitability with the experimental hardware and setup. At launch site, carrot seeds were placed between two disks of inert substrate (one imbibed with water and the other with a disodium phosphate solution) and integrated into a hardware developed, refurbished and flight-certificated by Kayser Italia. Post-flight, a Ground Reference Experiment was performed. Root development and orientation of seedlings grown in microgravity and at 1g condition were measured through 3D-image analysis procedures after imaging with X-ray microtomography. Radicle protruded preferentially from the ventral side of the seed due to the asymmetric position of the embryo. Such a phenomenon did not prevent the achievement of MULTITROP scientific goal but should be considered for further experiments on radicle growth orientation in microgravity. The experiment conducted in space verified that the primary root of carrot shows a positive chemotropism towards disodium phosphate solution in the absence of the gravity stimulus. On Earth, the positive chemotropism was masked by the dominant effect of gravity and roots developed downward regardless of the presence/absence of nutrients in the substrate. Taking advantage of altered gravity conditions and using other chemical compounds, further studies should be performed to deepen our understanding of root chemotropic response and its interaction with other tropisms.

Disruption of the Lotus japonicus transporter LjNPF2.9 increases shoot biomass and nitrate content without affecting symbiotic performances.

BACKGROUND: After uptake from soil into the root tissue, distribution and allocation of nitrate throughout the whole plant body, is a critical step of nitrogen use efficiency (NUE) and for modulation of plant growth in response to various environmental conditions. In legume plants nitrate distribution is also important for the regulation of the nodulation process that allows to fix atmospheric N (N2) through the symbiotic interaction with rhizobia (symbiotic nitrogen fixation, SNF). RESULTS: Here we report the functional characterization of the Lotus japonicus gene LjNPF2.9, which is expressed mainly in the root vascular structures, a key localization for the control of nitrate allocation throughout the plant body. LjNPF2.9 expression in Xenopus laevis oocytes induces 15NO3 accumulation indicating that it functions as a nitrate importer. The phenotypic characterization of three independent knock out mutants indicates an increased shoot biomass in the mutant backgrounds. This phenotype is associated to an increased/decreased nitrate content detected in the shoots/roots. Furthermore, our analysis indicates that the accumulation of nitrate in the shoot does not affect the nodulation and N-Fixation capacities of the knock out mutants. CONCLUSIONS: This study shows that LjNPF2.9 plays a crucial role in the downward transport of nitrate to roots, occurring likely through a xylem-to-phloem loading-mediated activity. The increase of the shoot biomass and nitrate accumulation might represent a relevant phenotype in the perspective of an improved NUE and this is further reinforced in legume plants by the reported lack of effects on the SNF efficiency.

Generalized EPID calibration for in vivo transit dosimetry.

Many researchers are studying new in vivo dosimetry methods based on the use of Elelctronic portal imaging devices (EPIDs) that are simple and efficient in their daily use. However the need of time consuming implementation measurements with solid water phantoms for the in vivo dosimetry implementation can discourage someone in their use. In this paper a procedure has been proposed to calibrate aSi EPIDs for in vivo transit dosimetry. The dosimetric equivalence of three aSi Varian EPIDs has been investigated in terms of signal reproducibility and long term stability, signal linearity with MU and dose per pulse and signal dependence on the field dimensions. The signal reproducibility was within ± 0.5% (2SD), while the long term signal stability has been maintained well within ± 2%. The signal linearity with the monitor units (MU) was within ± 2% and within ± 0.5% for the EPIDs controlled by the IAS 2, and IAS 3 respectively. In particular it was verified that the correction factor for the signal linearity with the monitor units, k(lin), is independent of the beam quality, and the dose per pulse absorbed by the EPID. For 6, 10 and 15 MV photon beams, a generalized set of correlation functions F(TPR,w,L) and empirical factors f(TPR,d,L) as a function of the Tissue Phantom Ratio (TPR), the phantom thickness, w, the square field side, L, and the distance, d, between the phantom mid-plane and the isocentre were determined to reconstruct the isocenter dose. The tolerance levels of the present in vivo dosimetry method ranged between ± 5% and ± 6% depending on the tumor body location. In conclusion, the procedure proposed, that use generalized correlation functions, reduces the effort for the in vivo dosimetry method implementation for those photon beams with TPR within ± 0.3% as respect those here used.

A generalized calibration procedure for in vivo transit dosimetry using siemens electronic portal imaging devices.

A practical and accurate generalized in vivo dosimetry procedure has been implemented for Siemens linacs supplying 6, 10, and 15 MV photon beams, equipped with aSi electronic portal imaging devices (EPIDs). The in vivo dosimetry method makes use of correlation ratios between EPID transit signal, s (t) (0) (TPR,w,L), and phantom mid-plane dose, D (0)(TPR,w,L), as functions of phantom thickness, w, square field dimensions, L, and tissue-phantom ratio TPR(20,10). The s (t) (0) (TPR,w,L) and D (0)(TPR,w,L) values were defined to be independent of the EPID sensitivity and monitor unit calibration, while their dependence on TPR(20,10) was investigated to determine a set of generalized correlation ratios to be used for beams with TPR(20,10) falling in the examined range. This way, other radiotherapy centers can use the method with no need to locally perform the whole set of measurements in solid water phantoms, required to implement it. Tolerance levels for 3D conformal treatments, ranging between ±5 and ±6% according to tumor type and location, were estimated for comparison purposes between reconstructed isocenter dose, D (iso), and treatment planning system (TPS) computed dose D (iso,TPS). Finally a dedicated software, interfaceable with record and verify (R&V) systems used in the centers, was developed to obtain in vivo dosimetry results in less than 2 min after beam delivery.

Correlation functions for Elekta aSi EPIDs used as transit dosimeter for open fields.

In-vivo dosimetry techniques are currently being applied only by a few Centers because they require time-consuming implementation measurements, and workload for detector positioning and data analysis. The transit in-vivo dosimetry performed by the electronic portal imaging device (EPID) avoids the problem of solid-state detector positioning on the patient. Moreover, the dosimetric characterization of the recent Elekta aSi EPIDs in terms of signal stability and linearity make these detectors useful for the transit in-vivo dosimetry with 6, 10 and 15 MV photon beams. However, the implementation of the EPID transit dosimetry requires several measurements. Recently, the present authors have developed an in-vivo dosimetry method for 3D CRT based on correlation functions defined by the ratios between the transit signal, st (w,L), by the EPID and the phantom midplane dose, Dm(w,L), at the source to axis distance (SAD) as a function of the phantom thickness, w, and the square field dimensions, L. When the phantom midplane was positioned at distance, d, from the SAD, the ratios st(w,L)/s't(d,w,L) were used to take into account the variation of the scattered photon contributions on the EPID as a function of d and L.The aim of this paper is the implementation of a procedure that uses generalized correlation functions obtained by nine Elekta Precise linac beams. The procedure can be used by other Elekta Precise linacs equipped with the same aSi EPIDs, assuming the stabilities of the beam output factors and the EPID signals. The procedure here reported avoids measurements in solid water equivalent phantoms needed to implement the in-vivo dosimetry method in the radiotherapy department. A tolerance level ranging between ± 5% and ± 6% (depending on the type of tumor) was estimated for the comparison between the reconstructed isocenter dose, Diso, and the computed dose, Diso,TPS, by the treatment planning system (TPS).