Effects of the Protein Kinase Inhibitor PKC412 on Gene Expression and Link to Physiological Effects in Zebrafish Danio rerio Eleuthero-Embryos

To identify molecular effects of the antineoplastic agent protein kinase C inhibitor 412 (PKC412) (midostaurin), we applied gene expression profiling in zebrafish using whole-genome microarrays. Behavioral, developmental, and physiological effects were investigated in order to analyze for correlations between altered gene expression profiles with effects on development and physiology. Zebrafish blastula-stage embryos were exposed for 6 days postfertilization to nominal levels of 2 and 40 μg/l PKC412. Among the 259 and 511 altered transcripts at both concentrations, respectively, the expressions of genes involved in the circadian rhythm were further investigated. Alteration of swimming behavior was not observed. Pathways of interest affected by PKC412 were angiogenesis, apoptosis, DNA damage response, and response to oxidative stress. Angiogenesis was analyzed in double-transgenic zebrafish embryos Tg(fli1a:EGFP)y1;Tg(gata1:dsRed)sd2; no major defects were induced by PKC412 treatment at both concentrations. Apoptosis occurred in olfactory placodes of embryos exposed to 40 μg/l, and DNA damage was induced at both PKC412 concentrations. However, there were no significant effects on reactive oxygen species formation. This study leads to the conclusion that PKC412-induced alterations of gene transcripts are partly paralleled by physiological effects at high, but not at low PKC412 concentrations expected to be of environmental relevanc

Protein interference applications in cellular and developmental biology using DARPins that recognize GFP and mCherry

Protein–protein interactions are crucial for cellular homeostasis and play important roles in the dynamic execution of biological processes. While antibodies represent a well-established tool to study protein interactions of extracellular domains and secreted proteins, as well as in fixed and permeabilized cells, they usually cannot be functionally expressed in the cytoplasm of living cells. Non-immunoglobulin protein-binding scaffolds have been identified that also function intracellularly and are now being engineered for synthetic biology applications. Here we used the Designed Ankyrin Repeat Protein (DARPin) scaffold to generate binders to fluorescent proteins and used them to modify biological systems directly at the protein level. DARPins binding to GFP or mCherry were selected by ribosome display. For GFP, binders with KD as low as 160 pM were obtained, while for mCherry the best affinity was 6 nM. We then verified in cell culture their specific binding in a complex cellular environment and found an affinity cut-off in the mid-nanomolar region, above which binding is no longer detectable in the cell. Next, their binding properties were employed to change the localization of the respective fluorescent proteins within cells. Finally, we performed experiments in Drosophila melanogaster and Danio rerio and utilized these DARPins to either degrade or delocalize fluorescently tagged fusion proteins in developing organisms, and to phenocopy loss-of-function mutations. Specific protein binders can thus be selected in vitro and used to reprogram developmental systems in vivo directly at the protein level, thereby bypassing some limitations of approaches that function at the DNA or the RNA level

Ocena przydatności parametrów przezklatkowego badania echokardiograficznego w diagnostyce choroby niedokrwiennej serca = The assessment of fitness parameters of transthoracic echocardiography in the diagnosis of coronary heart disease

Lenard Rafał, Zielińska Urszula, Michalczak Magdalena, Sobieszczańska Anna, Smoleń Agata. Ocena przydatności parametrów przezklatkowego badania echokardiograficznego w diagnostyce choroby niedokrwiennej serca = The assessment of fitness parameters of transthoracic echocardiography in the diagnosis of coronary heart disease. Journal of Education, Health and Sport. 2015;5(12):701-716. eISSN 2391-8306. DOI http://dx.doi.org/10.5281/zenodo.45069http://ojs.ukw.edu.pl/index.php/johs/article/view/2015%3B5%2812%29%3A701-716https://pbn.nauka.gov.pl/works/701302Formerly Journal of Health Sciences. ISSN 1429-9623 / 2300-665X. Archives 2011–2014http://journal.rsw.edu.pl/index.php/JHS/issue/archive     The journal has had 7 points in Ministry of Science and Higher Education parametric evaluation. Part B item 755 (23.12.2015).755 Journal of Education, Health and Sport (null) 2391-8306 7© The Author (s) 2015;This article is published with open access at Licensee Open Journal Systems of Kazimierz Wielki University in Bydgoszcz, Poland and Radom University in Radom, PolandOpen Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium,provided the original author(s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non Commercial License(http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited.This is an open access article licensed under the terms of the Creative Commons Attribution Non Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non commercialuse, distribution and reproduction in any medium, provided the work is properly cited.The authors declare that there is no conflict of interests regarding the publication of this paper.Received: 10.12.2015. Revised 25.12.2015. Accepted: 29.12.2015.  Ocena przydatności parametrów przezklatkowego badania echokardiograficznego w diagnostyce choroby niedokrwiennej serca The assessment of fitness parameters of transthoracic echocardiography in the diagnosis of coronary heart disease Rafał Lenard1, Urszula Zielińska2, Magdalena Michalczak1, Anna Sobieszczańska3, Agata Smoleń1 1Katedra i Zakład Epidemiologii i Metodologii Badań Klinicznych, Uniwersytet Medyczny w Lublinie2Klinika Neurochirurgii i Neurochirurgii Dziecięcej, Uniwersytet Medyczny w Lublinie3Katedra i Zakład Ortopedii Szczękowej, Uniwersytet Medyczny w Lublinie  StreszczenieWprowadzenieChoroby układu krążenia stanowią główną przyczynę zgonów w Europie. Przezklatkowe badanie echokardiograficzne TTE wykonuje się celem oceny anatomii serca, oceny kurczliwości globalnej i odcinkowej lewej komory, oceny aorty piersiowej oraz naczyń płucnych. TTE należy do podstawowych badań w diagnostyce choroby niedokrwiennej serca.Cel pracyCelem pracy była ocena przydatności parametrów przezklatkowego badania echokardiograficznego w diagnostyce zmian w naczyniach wieńcowych.Materiał i metodyBadaniem objęto 197 osób, w wieku od 43 do 88 lat, którzy byli hospitalizowani z powodu choroby niedokrwiennej serca (106 osób – 53,9% ze stabilną chorobą niedokrwienną serca, 91 osób – 46,1% z ostrym zespołem wieńcowym - OZW).Wyniki badańU pacjentów z OZW z obniżeniem frakcji wyrzutowej lewej komory częściej występowały istotne zmiany w tętnicach wieńcowych  (p<0,0001). Wśród  chorych z ostrym zespołem wieńcowym częściej występowała obniżona frakcja wyrzutowa lewej komory (62,6%) w porównaniu z pacjentami ze stabilną chorobą niedokrwienną serca (24,5%) (p<0,0001). W subpopulacji osób z OZW częściej występowały odcinkowe zaburzenia kurczliwości (67,0%) w porównaniu do grupy  osób ze stabilną chorobą niedokrwienną serca (8,0%) (p<0,0001).Wnioski1. Badanie echokardiograficzne powinno być wykonywane u każdego pacjenta z podejrzeniem choroby niedokrwiennej serca, jeżeli tylko nie opóźnia pilnej rewaskularyzacji mięśnia serca.2. Badanie echokardiograficzne jest najważniejszym badaniem w diagnostyce różnicowej bólu w klatce piersiowej oraz pozwala na stratyfikację ryzyka zdarzeń sercowo-naczyniowych.3. Do najważniejszych parametrów w diagnostyce ChNS należy ocena frakcji wyrzutowej oraz regionalnych zaburzeń kurczliwości mięśnia lewej komory.4. Zmniejszenie frakcji wyrzutowej oraz regionalne zaburzenia kurczliwości lewej komory są charakterystyczne dla zawału mięśnia serca (STEMI, NSTEMI). Słowa kluczowe: echokardiografia, choroba niedokrwienna serca, diagnostyka. AbstractIntroductionCardiovascular diseases are the major cause of death in Europe. Transthoracic echocardiography (TTE) is one of the most important examination in the diagnosis of coronary artery disease (CAD).Aim of the studyThe aim of the study was to evaluate parameters of TTE in the diagnosis of coronary artery disease.Methods and resultsThe study included 197 patients hospitalized with coronary artery disease (106 -53.9% with stable CAD, 91 – 46.1% with acute coronary syndrome - ACS). The major atherosclerotic changes in the coronary arteries were more frequent in males, in patients with ACS and reduction of left ventricular ejection fraction (p<0.0001). In patients with ACS left ventricular ejection fraction was more frequent decreased (62.6%) compared to patients with stable coronary artery disease (24.5%) (p <0,0001). In patients with ACS segmental wall motion abnormalities were more frequent 61/91 (67.0%) compared to patients with stable CAD 9/106 (8.0%) (p<0.0001)Conclusions:1. The echocardiography should be performed in every patient with suspected coronary heart disease, if only it doesn’t delay urgent revascularization of the heart muscle.2. The echocardiography is the most important examination in the differential diagnosis for the chest pain and allows the assessment of risk stratification for cardiovascular events.3. The most important parameter in the diagnosis of coronary heart disease is the assessment of the ejection fraction and regional wall motion abnormalities of the left ventricle.4. The reduction in ejection fraction and regional left ventricular wall motion abnormalities are characteristic for myocardial infarction (STEMI, NSTEMI). Keywords: echocardiography, coronary artery disease, diagnosis

Transapical Mitral Valve Replacement: 1-Year Results of the Real-World Tendyne European Experience Registry.

BACKGROUND Early studies of the Tendyne transcatheter mitral valve replacement (TMVR) showed promising results in a small selective cohort. OBJECTIVES The authors present 1-year data from the currently largest commercial, real-world cohort originating from the investigator-initiated TENDER (Tendyne European Experience) registry. METHODS All patients from the TENDER registry eligible for 1-year follow-up were included. The primary safety endpoint was 1-year cardiovascular mortality. Primary performance endpoint was reduction of mitral regurgitation (MR) up to 1 year. RESULTS Among 195 eligible patients undergoing TMVR (median age 77 years [Q1-Q3: 71-81 years], 60% men, median Society of Thoracic Surgeons Predicted Risk of Mortality 5.6% [Q1-Q3: 3.6%-8.9%], 81% in NYHA functional class III or IV, 94% with MR 3+/4+), 31% had "real-world" indications for TMVR (severe mitral annular calcification, prior mitral valve treatment, or others) outside of the instructions for use. The technical success rate was 95%. The cardiovascular mortality rate was 7% at 30 day and 17% at 1 year (all-cause mortality rates were 9% and 29%, respectively). Reintervention or surgery following discharge was 4%, while rates of heart failure hospitalization reduced from 68% in the preceding year to 25% during 1-year follow-up. Durable MR reduction to ≤1+ was achieved in 98% of patients, and at 1 year, 83% were in NYHA functional class I or II. There was no difference in survival and major adverse events between on-label use and "real-world" indications up to 1 year. CONCLUSIONS This large, real-world, observational registry reports high technical success, durable and complete MR elimination, significant clinical benefits, and a 1-year cardiovascular mortality rate of 17% after Tendyne TMVR. Outcomes were comparable between on-label use and "real-world" indications, offering a safe and efficacious treatment option for patients without alternative treatments. (Tendyne European Experience Registry [TENDER]; NCT04898335)

Numerical simulation of the impact of COVID-19 lockdown on tropospheric composition and aerosol radiative forcing in Europe

Aerosols influence the Earth\u27s energy balance directly by modifying the radiation transfer and indirectly by altering the cloud microphysics. Anthropogenic aerosol emissions dropped considerably when the global COVID-19 pandemic resulted in severe restraints on mobility, production, and public life in spring 2020. We assess the effects of these reduced emissions on direct and indirect aerosol radiative forcing over Europe, excluding contributions from contrails. We simulate the atmospheric composition with the ECHAM5/MESSy Atmospheric Chemistry (EMAC) model in a baseline (business-as-usual) and a reduced emission scenario. The model results are compared to aircraft observations from the BLUESKY aircraft campaign performed in May–June 2020 over Europe. The model agrees well with most of the observations, except for sulfur dioxide, particulate sulfate, and nitrate in the upper troposphere, likely due to a biased representation of stratospheric aerosol chemistry and missing information about volcanic eruptions. The comparison with a baseline scenario shows that the largest relative differences for tracers and aerosols are found in the upper troposphere, around the aircraft cruise altitude, due to the reduced aircraft emissions, while the largest absolute changes are present at the surface. We also find an increase in all-sky shortwave radiation of 0.21 ± 0.05 W m⁻² at the surface in Europe for May 2020, solely attributable to the direct aerosol effect, which is dominated by decreased aerosol scattering of sunlight, followed by reduced aerosol absorption caused by lower concentrations of inorganic and black carbon aerosols in the troposphere. A further increase in shortwave radiation from aerosol indirect effects was found to be much smaller than its variability. Impacts on ice crystal concentrations, cloud droplet number concentrations, and effective crystal radii are found to be negligible

Transapical mitral valve implantation for treatment of symptomatic mitral valve disease: a real-world multicentre experience.

AIMS Transcatheter mitral valve implantation (TMVI) is a new treatment option for patients with symptomatic mitral valve (MV) disease. Real-world data have not yet been reported. This study aimed to assess procedural and 30-day outcomes of TMVI in a real-world patient cohort. METHOD AND RESULTS All consecutive patients undergoing implantation of a transapically delivered self-expanding valve at 26 European centres from January 2020 to April 2021 were included in this retrospective observational registry. Among 108 surgical high-risk patients included (43% female, mean age 75 ± 7 years, mean STS-PROM 7.2 ± 5.3%), 25% was treated for an off-label indication (e.g. previous MV intervention or surgery, mitral stenosis, mitral annular calcification). Patients were highly symptomatic (New York Heart Association [NYHA] functional class III/IV in 86%) and mitral regurgitation (MR) was graded 3+/4+ in 95% (38% primary, 37% secondary, and 25% mixed aetiology). Technical success rate was 96%, and MR reduction to ≤1+ was achieved in all patients with successful implantation. There were two procedural deaths and 30-day all-cause mortality was 12%. At early clinical follow-up, MR reduction was sustained and there were significant reductions of pulmonary pressure (systolic pulmonary artery pressure 52 vs. 42 mmHg, p < 0.001), and tricuspid regurgitation severity (p = 0.013). Heart failure symptoms improved significantly (73% in NYHA class I/II, p < 0.001). Procedural success rate according to MVARC criteria was 80% and was not different in patients treated for an off-label indication (74% vs. 81% for off- vs. on-label, p = 0.41). CONCLUSION In a real-world patient population, TMVI has a high technical and procedural success rate with efficient and durable MR reduction and symptomatic improvement

Impact of reduced emissions on direct and indirect aerosol radiative forcing during COVID-19 lockdown in Europe

Aerosols influence the Earth’s energy balance through direct radiative effects and indirectly by altering the cloud microphysics. Anthropogenic aerosol emissions dropped considerably when the global COVID–19 pandemic resulted in severe restraints on mobility, production, and public life in spring 2020. Here we assess the effects of these reduced emissions on direct and indirect aerosol radiative forcing over Europe, excluding contributions from contrails. We simulate the atmospheric composition with the ECHAM5/MESSy Atmospheric Chemistry (EMAC) model in a baseline (business as usual) and a reduced emission scenario. The model results are compared to aircraft observations from the BLUESKY aircraft campaign performed in May June 2020 over Europe

Multidifferential study of identified charged hadron distributions in ZZ-tagged jets in proton-proton collisions at s=\sqrt{s}=13 TeV

Jet fragmentation functions are measured for the first time in proton-proton collisions for charged pions, kaons, and protons within jets recoiling against a $Z$ boson. The charged-hadron distributions are studied longitudinally and transversely to the jet direction for jets with transverse momentum 20 $< p_{\textrm{T}} < 100$ GeV and in the pseudorapidity range $2.5 < \eta < 4$. The data sample was collected with the LHCb experiment at a center-of-mass energy of 13 TeV, corresponding to an integrated luminosity of 1.64 fb$^{-1}$. Triple differential distributions as a function of the hadron longitudinal momentum fraction, hadron transverse momentum, and jet transverse momentum are also measured for the first time. This helps constrain transverse-momentum-dependent fragmentation functions. Differences in the shapes and magnitudes of the measured distributions for the different hadron species provide insights into the hadronization process for jets predominantly initiated by light quarks.Comment: All figures and tables, along with machine-readable versions and any supplementary material and additional information, are available at https://cern.ch/lhcbproject/Publications/p/LHCb-PAPER-2022-013.html (LHCb public pages

Study of the B−→Λc+Λˉc−K−B^{-} \to \Lambda_{c}^{+} \bar{\Lambda}_{c}^{-} K^{-} decay

The decay $B^{-} \to \Lambda_{c}^{+} \bar{\Lambda}_{c}^{-} K^{-}$ is studied in proton-proton collisions at a center-of-mass energy of $\sqrt{s}=13$ TeV using data corresponding to an integrated luminosity of 5 $\mathrm{fb}^{-1}$ collected by the LHCb experiment. In the $\Lambda_{c}^+ K^{-}$ system, the $\Xi_{c}(2930)^{0}$ state observed at the BaBar and Belle experiments is resolved into two narrower states, $\Xi_{c}(2923)^{0}$ and $\Xi_{c}(2939)^{0}$, whose masses and widths are measured to be $$m(\Xi_{c}(2923)^{0}) = 2924.5 \pm 0.4 \pm 1.1 \,\mathrm{MeV}, \\ m(\Xi_{c}(2939)^{0}) = 2938.5 \pm 0.9 \pm 2.3 \,\mathrm{MeV}, \\ \Gamma(\Xi_{c}(2923)^{0}) = \phantom{000}4.8 \pm 0.9 \pm 1.5 \,\mathrm{MeV},\\ \Gamma(\Xi_{c}(2939)^{0}) = \phantom{00}11.0 \pm 1.9 \pm 7.5 \,\mathrm{MeV},$$ where the first uncertainties are statistical and the second systematic. The results are consistent with a previous LHCb measurement using a prompt $\Lambda_{c}^{+} K^{-}$ sample. Evidence of a new $\Xi_{c}(2880)^{0}$ state is found with a local significance of $3.8\,\sigma$, whose mass and width are measured to be $2881.8 \pm 3.1 \pm 8.5\,\mathrm{MeV}$ and $12.4 \pm 5.3 \pm 5.8 \,\mathrm{MeV}$, respectively. In addition, evidence of a new decay mode $\Xi_{c}(2790)^{0} \to \Lambda_{c}^{+} K^{-}$ is found with a significance of $3.7\,\sigma$. The relative branching fraction of $B^{-} \to \Lambda_{c}^{+} \bar{\Lambda}_{c}^{-} K^{-}$ with respect to the $B^{-} \to D^{+} D^{-} K^{-}$ decay is measured to be $2.36 \pm 0.11 \pm 0.22 \pm 0.25$, where the first uncertainty is statistical, the second systematic and the third originates from the branching fractions of charm hadron decays.Comment: All figures and tables, along with any supplementary material and additional information, are available at https://cern.ch/lhcbproject/Publications/p/LHCb-PAPER-2022-028.html (LHCb public pages