Search

Abstract Kindlin-2 is a cytoskeletal adapter protein that is present in many different cell types. By virtue of its interaction with multiple binding partners, Kindlin-2 intercalates into numerous signaling pathways and cytoskeletal nodes. A specific interaction of Kindlin-2 that is of paramount importance in many cellular responses is its direct binding to the cytoplasmic tails of integrins, an interaction that controls many of the adhesive, migratory and signaling responses mediated by members of the integrin family of cell-surface heterodimers. Kindlin-2 is highly expressed in many cancers and is particularly prominent in prostate cancer cells. CRISPR/cas9 was used as a primary approach to knockout expression of Kindlin-2 in both androgen-independent and dependent prostate cancer cell lines, and the effects of Kindlin-2 suppression on oncogenic properties of these prostate cancer cell lines was examined. Adhesion to extracellular matrix proteins was markedly blunted, consistent with the control of integrin function by Kindlin-2. Migration across matrices was also affected. Anchorage independent growth was markedly suppressed. These observations indicate that Kindlin-2 regulates hallmark features of prostate cancer cells. In androgen expressing cells, testosterone-stimulated adhesion was Kindlin-2-dependent. Furthermore, tumor growth of a prostate cancer cell line lacking Kindlin-2 and implanted into the prostate gland of immunocompromised mice was markedly blunted and was associated with suppression of angiogenesis in the developing tumor. These results establish a key role of Kindlin-2 in prostate cancer progression and suggest that Kindlin-2 represents an interesting therapeutic target for treatment of prostate cancer.

Abstract Background The current generation of radiolabeled PSMA-targeting therapeutic agents is limited by prominent salivary gland binding, which results in dose-limiting xerostomia from radiation exposure. JB-1498 is a urea-based small molecule with a highly negatively charged linker targeting prostate specific membrane antigen (PSMA). Prior work on a similar tracer with the same negatively charged linker demonstrated low normal organ/soft tissue background uptake compared to [68Ga]Ga-PSMA-11. The purpose of this study was to investigate if [68Ga]Ga-JB-1498 had reduced salivary gland uptake in mice compared to [68Ga]Ga-PSMA-11. Results JB-1498 demonstrated high affinity for PSMA binding and tumor uptake in a murine tumor model. In an initial biodistribution study with low molar activity, [68Ga]Ga-JB-1498 demonstrated salivary gland uptake of 0.13 ± 0.01%ID/g. In a second biodistribution study in non-tumor-bearing mice with high molar activity, [68Ga]Ga-JB1498 demonstrated salivary gland uptake of 0.39 ± 0.24% ID/g and kidney activity of 10.12 ± 1.73% ID/g at one hour post IV injection. This salivary gland uptake is significantly less than the published uptake of [68Ga]Ga-PSMA-11. Micro-PET visually confirmed the findings of the biodistribution studies. Dynamic micro-PET imaging demonstrated gradually decreasing [68Ga]Ga-JB1498 activity in salivary glands and kidneys, compared to gradually increasing [68Ga]Ga-PSMA-11 activity in these two organs during the first hour. Conclusion Biodistribution and micro-PET imaging of [68Ga]Ga-JB-1498 demonstrate significantly decreased salivary gland uptake and different pharmacokinetic behavior in kidneys and salivary glands in mice compared to [68Ga]Ga-PSMA-11. Our findings suggest that constructing a PSMA-targeting molecule with a highly negatively charged linker is a promising strategy to reduce salivary gland uptake of GCP-II/PSMA ligands in theranostic applications.

Summary: Small cell lung cancers (SCLCs) rapidly resist cytotoxic chemotherapy and immune checkpoint inhibitor (ICI) treatments. New, non-cross-resistant therapies are thus needed. SCLC cells are committed into neuroendocrine lineage then maturation arrested. Implicating DNA methyltransferase 1 (DNMT1) in the maturation arrests, we find (1) the repression mark methylated CpG, written by DNMT1, is retained at suppressed neuroendocrine-lineage genes, even as other repression marks are erased; (2) DNMT1 is recurrently amplified, whereas Ten-Eleven-Translocation 2 (TET2), which functionally opposes DNMT1, is deleted; (3) DNMT1 is recruited into neuroendocrine-lineage master transcription factor (ASCL1, NEUROD1) hubs in SCLC cells; and (4) DNMT1 knockdown activated ASCL1-target genes and released SCLC cell-cycling exits by terminal lineage maturation, which are cycling exits that do not require the p53/apoptosis pathway used by cytotoxic chemotherapy. Inhibiting DNMT1/corepressors with clinical compounds accordingly extended survival of mice with chemorefractory and ICI-refractory, p53-null, disseminated SCLC. Lineage commitment of SCLC cells can hence be leveraged into non-cytotoxic therapy able to treat chemo/ICI-refractory SCLC.

The rise of Fe magnetic moment, changes in Al electronic structure and a variation of Al magnetic polarization in thin films of transition metal aluminide Fe _60 Al _40 have been probed through the order-disorder phase transition by soft x-ray absorption spectroscopy and x-ray resonant magnetic reflectivity in the extreme ultraviolet regime. In a course of the transition induced by 20 keV Ne ^+ irradiation with low fluences (∼10 ^14 ions cm ^−2 ), x-ray magnetic circular dichroism spectra taken at the Fe L $_{2,3}$ absorption edges at room and low temperatures revealed a pronounced increase of Fe $3\textit{d}$ states spin-polarization. X-ray resonant magnetic reflectivity applied to the Al L $_{2,3}$ and Fe M $_{2,3}$ edges allowed to detect the magnetic polarization of Al atoms in the films. The changes in Al electronic structure have been seen by alteration of Al K edge x-ray absorption near edge structure. A difference in anisotropy fields for films before and after irradiation has been observed by element-specific hysteresis loops recorded at low temperatures in absorption and reflection geometries at the Fe L $_{2,3}$ and M $_{2,3}$ edges, respectively. An attempt to reduce the top oxide layer by an inductively coupled hydrogen plasma has shown a possibility to recover the chemically ordered phase.

In our recent work [Körber et al., AIP Adv. 11, 095006 (2021)], we presented an efficient numerical method to compute dispersions and mode profiles of spin waves in waveguides with translationally invariant equilibrium magnetization. A finite-element method (FEM) allowed to model two-dimensional waveguide cross sections of arbitrary shape but only finite size. Here, we extend our FEM propagating-wave dynamic-matrix approach from finite waveguides to the important cases of infinitely extended mono- and multi-layers of arbitrary spacing and thickness. To obtain the mode profiles and frequencies, the linearized equation of the motion of magnetization is solved as an eigenvalue problem on a one-dimensional line-trace mesh, defined along the normal direction of the layers. Being an important contribution to multi-layer systems, we introduce interlayer exchange into our FEM approach. With the calculation of dipolar fields being the main focus, we also extend the previously presented plane-wave Fredkin–Koehler method to calculate the dipolar potential of spin waves in infinite layers. The major benefit of this method is that it avoids the discretization of any non-magnetic material such as non-magnetic spacers in multilayers. Therefore, the computational effort becomes independent of the spacer thicknesses. Furthermore, it keeps the resulting eigenvalue problem sparse, which, therefore, inherits a comparably low arithmetic complexity. As a validation of our method (implemented into the open-source finite-element micromagnetic package TETRAX), we present results for various systems and compare them with theoretical predictions and with established finite-difference methods. We believe this method offers an efficient and versatile tool to calculate spin-wave dispersions in layered magnetic systems.

The ketogenic diet (KD) is hypothesized to impact tumor progression by altering tumor metabolism. In this study, we assessed the impact of an unrestricted KD on epithelial ovarian cancer (EOC) tumor growth, gene expression, and metabolite concentration in a mouse model. ID8 EOC cells, which were syngeneic with C57Bl/6J mouse strain and transfected with luciferase (ID8-luc), were injectedand monitored for tumor development. Female mice were fed either a strict KD, a high fat/low carbohydrate (HF/LC) diet, or a low fat/high carbohydrate (LF/HC) diet (n = 10 mice per group) ad libitum. EOC tumor growth was monitored weekly, and tumor burden was determined based on luciferase fluorescence (photons/second). At the endpoint (42 days), tumors were collected and processed for RNA sequencing. Plasma and tumor metabolites were evaluated using LC-MS. The KD-fed mice exhibited a statistically significant increase in tumor progression in comparison to the HF/LC- and LF/HC-fed groups (9.1 vs. 2.0 vs. 3.1-fold, respectively, p < 0.001). The EOC tumors of the KD-fed mice exhibited significant enrichment of the peroxisome proliferator-activated receptor (PPAR) signaling and fatty acid metabolism pathways based on the RNA sequencing analysis when compared to the LF/HC- and HF/LC-fed mice. Thus, unrestricted KD diet enhanced tumor progression in our mouse EOC model. KD was associated with the upregulation of fatty acid metabolism and regulation pathways, as well as enrichment of fatty acid and glutamine metabolites.

Self-determination theory posits that relatedness and autonomy are two drivers of work-relevant outcomes. Through the lens of this theory, the current study explored the potential interactive effects of relatedness and autonomy on affective, relational, and behavioral outcomes at work, operationalizing relatedness as workplace loneliness and autonomy as the ability to work from home. To test this relation, survey-based data from a sample of 391 working adults were collected and a path analysis was carried out. Results suggested that workplace loneliness negatively predicts affective organizational commitment, perceptions of coworker and supervisor support, organizational citizenship behaviors, and perceived performance. Furthermore, results suggested that workplace loneliness and working from home have an interactive effect on affective organizational commitment, perceptions of coworker support, and organizational citizenship behaviors. Specifically, working from home had a beneficial impact on the relation between workplace loneliness and affective organizational commitment/perceptions of coworker support, but a detrimental impact on the relation between workplace loneliness and organizational citizenship behaviors. These results add to the extant body of scholarly work of Self-Determination Theory by testing the theory in the post-pandemic context of working from home. In addition, these results have practical implications for managers, who should strive to create opportunities for employees who work from home to enact organizational citizenship behaviors.

Spin pumping from a metallic ferromagnet (FM) into an insulating antiferromagnet has been studied across the magnetic phase transition by means of temperature-dependent, broad-band ferromagnetic resonance (FMR) experiments. A set of spin pumping heterostructures consisting of Permalloy (Ni _80 Fe _20 ) as FM and Zn $_{1-x}$ Co _x O with $x = 0.3, 0.5$ and 0.6 (Co:ZnO) as partially compensated antiferromagnetic insulator has been used for which previous experiments have already pointed out the possibility of the existence of spin-pumping. The present experiments allow to reliably separate the various contributions of the temperature-dependent Gilbert damping parameter to the FMR line-width. A careful analysis of the obtained data demonstrates a significant increase of the temperature-dependence of the Gilbert damping parameter $\alpha(T)$ around the magnetic phase transition of Co:ZnO which extends up to room temperature, confirming spin pumping into the fluctuating spin sink of an antiferromagnetic/paramagnetic insulator.

This year marks the 20th anniversary of the launch of the chronic thromboembolic pulmonary hypertension (CTEPH) surgical treatment programme, which involves three departments of the Cardiac Centre of the General University Hospital in Prague. Diagnosis and decision-making on the type of treatment, including indication for surgical treatment, is carried out at the Centre. Other non-surgical treatments and long-term dispensary care are also available to patients. Over 500 patients with CTEPH have been operated on during the program's lifetime. Our Centre also concentrates on patients with other types of severe pulmonary hypertension who are indicated for various challenging surgical procedures, as they can be offered comprehensive perioperative care by experienced anaesthetists and intensivists. Anaesthesia and intensive postoperative care of patients indicated for pulmonary endarterectomy has certain specifics, which are based on the fact that patients undergoing this operation have right-sided heart failure with chronic low cardiac output, are significantly symptomatic, and have minimal circulatory and ventilatory reserves. In addition, they are at risk of developing several serious complications during the surgery and in the early postoperative period, which increase morbidity and mortality. This review work is based on twenty years of experience at our center, many of which have been materialized in independent original studies published in peer-reviewed journals. In addition to their practice, the authors draw on the experience of other European and international centers, many of which they have established useful collaborations with. The anaesthetic and intensive care procedures used have undergone many changes over the past twenty years and have been re-evaluated based on recommendations and experience, always to improve outcomes and reduce the rate of perioperative complications. [ABSTRACT FROM AUTHOR]

Harvesting ohmic heat for signal processing is one of major challenges in modern electronics and spin caloritronics, but not yet well accomplished. Here the authors demonstrate a spin torque oscillator device driven by pure spin current arising from thermal gradient across an Y3Fe5O12/Pt interface.

A theoretical approach has been developed to study the spin-wave dynamics of magnetization-graded ferromagnetic films, where the magnetic properties change along the film thickness. The theory is based on a multilayer approach, where the influence of both long-range dipolar interactions and interlayer exchange coupling between sublayers is included. This allows for instance to describe films with a continuous variation of the saturation magnetization along the thickness. A systematic study is carried out in order to analyze different profiles of the saturation magnetization, which is checked through a test of convergence. It is found that the spin-wave dispersion is significantly modified when the strength of the magnetization changes in the bulk film, where a notable frequency non-reciprocity of two counter propagating spin waves is predicted. This is associated with heterosymmetric mode profiles and a modification of the conventional quantization condition associated to perpendicular standing spin-wave modes. Micromagnetic simulations have been carried out to validate the model, where a perfect agreement is reached between both methods. These results show that magnetization-graded ferromagnetic films can be used to channel and control spin waves, thus promoting different kinds of functionalities for magnon-based devices.

The incidence of post infarction ventricular septal rupture (PIVSR) is decreasing in the last years due to aggressive treatment of myocardial infarction with early percutaneous coronary interventions. As a consequence patients with PIVSR are referred to surgery more often with significant heart failure. The aim of this retrospective study was to assess the influence of these on the operative results and to identify the risk factors of operative mortality. A retrospective analysis of prospectively collected data of patients with the PIVSR admitted to our center from November 2004 to February 2012 was performed. Variables were analyzed using two-dimensional correspondence analysis. There were 25 patients (12 males and 13 females) with mean age 70.2 years (47–82) operated on; 17 (68%) presented with anterior and 8 (32%) with posterior PIVSR. Eighteen patients (72%) had acute heart failure, 13 (52%) presented with cardiogenic shock. Before surgery, intraaortic balloon pump (IABP) had 20 (80%) patients; in 4 (16%) a ventricular assist device was used, either Extracorporeal Membrane Oxygenation (ECMO) or centrifugal pumps as biventricular assist. Operative mortality was 40% (10 pts.). Four patients (12%) had small non-significant recurrent shunt on postoperative echocardiography. Although majority of patients with PIVSR have significant heart failure prior to surgery the operative mortality remains comparable to older studies. Predictors of perioperative death were concomitant surgical reconstruction of the left ventricle, renal impairment before operation, male gender, history of coronary artery disease, PIVSR location posterior, and shock at surgery.

The authors present a case of massive lung bleeding following pulmonary endarterectomy (PEA) that was treated with peripheral veno-venous extracorporeal membrane oxygenation (VV ECMO). The patient repeatedly underwent bronchoscopy for airway blood clot obstruction and finally was successfully weaned off the support. The authors discuss the indications for ECMO in treatment of the most serious complications following PEA, and emphasize the importance of echocardiographic evaluation of the right ventricular function in relation to the indicated type of extracorporeal support. Anticoagulation strategy for patients shortly after the major surgery connected to ECMO is also discussed.

During tumor progression, EphA2 receptor can gain ligand-independent pro-oncogenic functions due to Akt activation and reduced ephrin-A ligand engagement. The effects can be reversed by ligand stimulation, which triggers the intrinsic tumor suppressive signaling pathways of EphA2 including inhibition of PI3/Akt and Ras/ERK pathways. These observations argue for development of small molecule agonists for EphA2 as potential tumor intervention agents. Through virtual screening and cell-based assays, we report here the identification and characterization of doxazosin as a novel small molecule agonist for EphA2 and EphA4, but not for other Eph receptors tested. NMR studies revealed extensive contacts of doxazosin with EphA2/A4, recapitulating both hydrophobic and electrostatic interactions recently found in the EphA2/ephrin-A1 complex. Clinically used as an α1-adrenoreceptor antagonist (Cardura®) for treating hypertension and benign prostate hyperplasia, doxazosin activated EphA2 independent of α1-adrenoreceptor. Similar to ephrin-A1, doxazosin inhibited Akt and ERK kinase activities in an EphA2-dependent manner. Treatment with doxazosin triggered EphA2 receptor internalization, and suppressed haptotactic and chemotactic migration of prostate cancer, breast cancer, and glioma cells. Moreover, in an orthotopic xenograft model, doxazosin reduced distal metastasis of human prostate cancer cells and prolonged survival in recipient mice. To our knowledge, doxazosin is the first small molecule agonist of a receptor tyrosine kinase that is capable of inhibiting malignant behaviors in vitro and in vivo.

Nitrosylcobalamin (NO-Cbl) is a chemotherapeutic pro-drug derived from vitamin B12 that preferentially delivers nitric oxide (NO) to tumor cells, based upon increased receptor expression. NO-Cbl induces Apo2L/TRAIL-mediated apoptosis and inhibits survival signaling in a variety of malignant cell lines. Chemotherapeutic agents often simultaneously induce an apoptotic signal and activation of NF-kappaB, which has the undesired effect of promoting cell survival. The specific aims of this study were to 1) measure the anti-tumor effects of NO-Cbl alone and in combination with conventional chemotherapeutic agents, and to 2) examine the mechanism of action of NO-Cbl as a single agent and in combination therapy.Using anti-proliferative assays, electrophoretic mobility shift assay (EMSA), immunoblot analysis and kinase assays, we demonstrate an increase in the effectiveness of chemotherapeutic agents in combination with NO-Cbl as a result of suppressed NF-kappaB activation.Eighteen chemotherapeutic agents were tested in combination with NO-Cbl, in thirteen malignant cell lines, resulting in a synergistic anti-proliferative effect in 78% of the combinations tested. NO-Cbl pre-treatment resulted in decreased NF-kappaB DNA binding activity, inhibition of IkappaB kinase (IKK) enzymatic activity, decreased AKT activation, increased caspase-8 and PARP cleavage, and decreased cellular XIAP protein levels.The use of NO-Cbl to inhibit survival signaling may enhance drug efficacy by preventing concomitant activation of NF-kappaB or AKT.

A joint theoretical–experimental study focusing on the description of the ferromagnetic resonance response of thin films in the presence of periodic perturbations introduced on the upper film surface is presented. From the viewpoint of theory, these perturbations may exist in the form of any kind of one- or two-dimensional rectangular defect arrays patterned onto one surface of the magnetic film. Indeed, the defects may be pits or bumps, or ion-implanted regions with a lower saturation magnetization. The complete set of response functions, given by the components of the frequency and wave-vector dependent dynamic magnetic susceptibility tensor of the film exposed to microwave excitation, are provided and are used to explain the experimental data. This allows us to obtain the response of the system due to microwave absorption, from which the zero wave-vector spin-wave modes in the field-frequency spectra, including their intensity, are calculated. Explicit calculations for periodic defects featuring the shape of stripes, dots and rectangles are given in detail, as well as experimental results for stripe-like defects prepared either by topographical depressions or by ion implantation of thin magnetic films. The excellent agreement of the theoretical and experimental results manifests the validity of the presented model.

OBJECTIVES: To compare clinical parameters and quality of life in patients with pulmonary arterial hypertension (PAH) at the time of diagnosis, at the time of LenusPro pump implantation and during intravenous treptostinil treatment. METHODS: Seven patients with severe PAH treated with intravenous treptostinil via implantable LenusPro pumps were evaluated, including NYHA classification, six-minute walking test, BNP and quality of life assessment using the EQ-5D-5L questionnaire before and after pump implantation. RESULTS: No significant changes were observed in NYHA class and six-minute walking distance test. There was however a significant improvement in the quality of life and a decrease in BNP levels. The mean EQ-5D- 5L index assessed during subcutaneous treptostinil treatment was significantly worse when compared to that assessed during its intravenous application (0.39 ± 0.24 vs 0.78 ± 0.28, p < 0.05); the same is true about the pain/discomfort dimension. Complications occurred, namely one nonfatal pneumothorax, one nonfatal hemothorax, and one event of nonfatal treptostinil intoxication after refilling. CONCLUSIONS: In patients who do not tolerate subcutaneous treptostinil treatment, the use of the LenusPro implantable pump results in a significant improvement in quality of life with an acceptable safety profile (Tab. 2, Fig. 2, Ref. 19). Text in PDF www.elis.sk. [ABSTRACT FROM AUTHOR]

OBJECTIVES: The aim of this study was to analyse survival of patients with pulmonary arterial hypertension (PAH) and chronic thromboembolic pulmonary hypertension (CTEPH) hospitalized due to an acute right heart failure (ARHF) with emphasis on risk factors and effectiveness of treatment following current guidelines. METHODS: We retrospectively analysed 117 hospitalizations of 70 patients (59 PAH patients; 11 CTEPH patients, mean age 53.1 ± 16.77 years, 54% females) between 2004 and 2013. RESULTS: 96 cases were hospitalized at cardiology wards (CW) while 21 at intensive care unit (ICU). The overall hospital mortality was 12.8%, CW mortality was 4%, and ICU mortality was 52.4%. Higher risk of in-hospital mortality was associated with younger age, lower sodium levels, severe forms of PAH (heritable PAH, CTD-PAH) and need of PAH combination treatment. The one-year survival from the fi rst ARHF hospitalization was 67.6% (95% CI 57.1-80%), the two-year survival was 41.9% (95% CI 30.8-56.9%). The presence of ascites was a predictor of long-term mortality. CONCLUSIONS: Mortality in patients with PH and ARHF remains very high. Identification of its risk factors could be used as basis of risk-adapted therapy (Tab. 5, Fig. 2, Ref. 14). Text in PDF www.elis.sk. [ABSTRACT FROM AUTHOR]

Interferons (IFNs) have proven antitumor activity against a variety of human malignancies, which may result, at least in part, from inhibition of angiogenesis. The objective of this study was to identify IFN-stimulated genes (ISGs) that played a role in mediation of angiogenic inhibition. IFN- was a more potent antiangiogenic agent compared to IFN-2b (80% versus 20%, respectively) and suggests that IFNs inhibited angiogenesis by preventing endothelial cell differentiation, and not by direct antiproliferative effects. To identify ISGs that were key inhibitors of angiogenesis, we utilized an in vitrofibrin gel angiogenic assay which closely recapitulated the in vivoprocesses of angiogenesis. DNA microarray analysis of IFN--treated endothelial cells in the fibrin gel assay identified 11 ISGs that were induced >10-fold during angiogenesis inhibition. Recombinant IP-10 inhibited angiogenesis in a dose-dependent fashion, but was a less effective inhibitor compared to IFN-, suggesting that additional ISGs are involved in inhibiting angiogenesis. ISG20 was upregulated by microarray analysis, but did not inhibit angiogenesis when overexpressed in human umbilical vein endothelial cells (HUVECs). However, a dominant negative mutant of ISG20 inhibited angiogenesis by 43%. Results suggest that IFN-induced angiogenic inhibition was likely mediated by multiple ISGs; our novel finding is that decreased exonuclease activity in HUVECs associated with expression of the ISG20 ExoII mutant inhibited angiogenesis. [ABSTRACT FROM AUTHOR]

Similar to electrical currents flowing through magnetic multilayers, thermal gradients applied across the barrier of a magnetic tunnel junction may induce pure spin-currents and generate ‘thermal’ spin-transfer torques large enough to induce magnetization dynamics in the free layer. In this study, we describe a novel experimental approach to observe spin-transfer torques induced by thermal gradients in magnetic multilayers by studying their ferromagnetic resonance response in microwave cavities. Utilizing this approach allows for measuring the magnetization dynamics on micron/nano-sized samples in open-circuit conditions, i.e. without the need of electrical contacts. We performed first experiments on magnetic tunnel junctions patterned into 6  ×  9 µm2 ellipses from Co2FeAl/MgO/CoFeB stacks. We conducted microresonator ferromagnetic resonance (FMR) under focused laser illumination to induce thermal gradients in the layer stack and compared them to measurements in which the sample was globally heated from the backside of the substrate. Moreover, we carried out broadband FMR measurements under global heating conditions on the same extended films the microstructures were later on prepared from. The results clearly demonstrate the effect of thermal spin-torque on the FMR response and thus show that the microresonator approach is well suited to investigate thermal spin-transfer-driven processes for small temperatures gradients, far below the gradients required for magnetic switching. [ABSTRACT FROM AUTHOR]

We demonstrate the excitation of the gyrotropic mode in a magnetostrictive vortex by time-varying strain. The vortex dynamics is driven by a time-varying voltage applied to the piezoelectric substrate and detected electrically by spin rectification at subthreshold values of rf current. When the frequency of the time-varying strain matches the gyrotropic frequency at a given in-plane magnetic field, the strain-induced in-plane magnetic anisotropy leads to a resonant excitation of the gyration dynamics in the magnetic vortex. We show that nonlinear gyrotropic dynamics can be excited already for moderate amplitudes of the time-varying strain.

Rare genetic variants can have strong effects on phenotypes, yet accounting for rare variants in genetic analyses is statistically challenging due to the limited number of allele carriers and the burden of multiple testing. While rich variant annotations promise to enable well-powered rare variant association tests, methods integrating variant annotations in a data-driven manner are lacking. Here we propose deep rare variant association testing (DeepRVAT), a model based on set neural networks that learns a trait-agnostic gene impairment score from rare variant annotations and phenotypes, enabling both gene discovery and trait prediction. On 34 quantitative and 63 binary traits, using whole-exome-sequencing data from UK Biobank, we find that DeepRVAT yields substantial gains in gene discoveries and improved detection of individuals at high genetic risk. Finally, we demonstrate how DeepRVAT enables calibrated and computationally efficient rare variant tests at biobank scale, aiding the discovery of genetic risk factors for human disease traits., (© 2024. The Author(s).)

Purpose: The PI3K signaling pathway is frequently dysregulated in breast cancer, and mutations in PIK3CA are relevant for therapy resistance in HER2-positive (HER2pos) breast cancer. Mutations in exons 9 or 20 may have different impacts on response to neoadjuvant chemotherapy-based treatment regimens., Experimental Design: We investigated PIK3CA mutations in 1,691 patients with early breast cancer who were randomized into four neoadjuvant multicenter trials: GeparQuattro (NCT00288002), GeparQuinto (NCT00567554), GeparSixto (NCT01426880), and GeparSepto (NCT01583426). The role of different PIK3CA exons and hotspots for pathologic complete response (pCR) following neoadjuvant chemotherapy (NACT) and patient survival were evaluated for distinct molecular subgroups and anti-HER2 treatment procedures., Results: A total of 302 patients (17.9%) of the full cohort of 1,691 patients had a tumor with a PIK3CA mutation, with a different prevalence in molecular subgroups: luminal/HER2-negative (HER2neg) 95 of 404 (23.5%), HER2pos 170 of 819 (20.8%), and triple-negative breast cancer 37 of 468 patients (7.9%). We identified the mutations in PIK3CA exon 20 to be linked with worse response to anti-HER2 treatment (OR = 0.507; 95% confidence interval, 0.320-0.802; P = 0.004), especially in hormone receptor-positive HER2-positive breast cancer (OR = 0.445; 95% confidence interval, 0.237-0.837; P = 0.012). In contrast, exon 9 hotspot mutations p.E452K and p.E545K revealed no noteworthy differences in response therapy. Luminal/HER2neg patients show a trend to have worse treatment response when PIK3CA was mutated. Interestingly, patients with residual disease following neoadjuvant treatment had better survival rates when PIK3CA was mutated., Conclusions: The PIK3CA hotspot mutation p.H1047R is associated with worse pCR rates following NACT in HER2pos breast cancer, whereas hotspot mutations in exon 9 seem to have less impact., (©2024 American Association for Cancer Research.)

In this Letter, we demonstrate terahertz (THz) magnetic field detection in fused silica with sensitivity that can be easily controlled by sample tilting (for both amplitude and polarization). The proposed technique remains in the linear regime at magnetic fields exceeding 0.3 T (0.9 MV/cm of equivalent electric field) and allows the use of low-cost amorphous materials. Furthermore, the demonstrated effects should be present in a wide variety of materials used as substrates in different THz-pump laser-probe experiments and need to be considered in order to disentangle different contributions to the measured signals.

An analytical method for studying DNA degradation by electrophoresis after cell lysis and visualization of DNA fragments with fluorescent dye, comet assay, was used to evaluate the viability of the endothelial layer of human arterial grafts with the aim of identifying the procedure that will least damage the tissue before cryopreservation. Four groups of samples were studied: cryopreserved arterial grafts that were thawed in two different ways, slowly lasting 2 hours or rapidly for approx. 7 minutes. Arterial grafts that were collected as part of multiorgan procurement with minimal warm ischemia time. Cadaveric grafts were taken as part of the autopsy, so they have a more extended period of warm ischemia. The HeadDNA (%) parameter and others commonly used parameters like TailDNA (%). TailMoment, TailLength, OliveMoment, TailMoment to characterize the comet were used to assess viability in this study. The ratio of non-decayed to decayed nuclei was determined from the values found. This ratio for cadaveric grafts was 0.63, for slowly thawed cryopreserved grafts 2.9, for rapidly thawed cryopreserved grafts 1.9, and for multi-organ procurement grafts 0.68. The results of the study confirmed the assumption that the allografts obtained from cadaveric donors are the least suitable. On the other hand, grafts obtained from multiorgan donors are better in terms of viability monitored by comet assay. Keywords: Arterial grafts, Cryopreservation, Cadaveric, Multiorgan procurement, Viability, Comet assay.

A case series of the use of amniotic membrane (AM) for treating chronic nonhealing wounds. It presents five cases of polymorbid patients with a total of nine chronic nonhealing wounds. The patient group consisted of four men and one woman with various comorbidities, aged 45-72 years. The mean initial wound size was 15.8 cm 2 , and the mean time from the onset of the wound to the first application of AM was 122 weeks. The wounds were caused by chronic venous insufficiency and/or peripheral arterial disease. Wounds were treated in a standardized protocol. AM was applied weekly in the first month and then every two weeks. Photo documentation of the wound and microbiological colonization was carried out at each visit. In three out of five patients, the AM treatment effectively promoted healing up to complete wound closure. In two cases, the wounds stayed unhealed despite numerous AM applications. Pain relief was noted in all patients. The success of the treatment was closely tied to patient factors, such as adherence to the prescribed treatment regimen and individual patient characteristics. In some cases, treatment failure was observed, possibly due to underlying comorbidities, wound parameters, or poor patient compliance. AM treatment has the potential to become a viable treatment option for these nonhealing wounds. However, the effectiveness of the treatment may be influenced by various patient factors and the underlying cause of the wound. Therefore, it is crucial to have an individualized treatment plan that considers these particular factors., (© 2023. The Author(s).)

Mitochondrial quality control failure is frequently observed in neurodegenerative diseases. The detection of damaged mitochondria by stabilization of PTEN-induced kinase 1 (PINK1) requires transport of Pink1 messenger RNA (mRNA) by tethering it to the mitochondrial surface. Here, we report that inhibition of AMP-activated protein kinase (AMPK) by activation of the insulin signalling cascade prevents Pink1 mRNA binding to mitochondria. Mechanistically, AMPK phosphorylates the RNA anchor complex subunit SYNJ2BP within its PDZ domain, a phosphorylation site that is necessary for its interaction with the RNA-binding protein SYNJ2. Notably, loss of mitochondrial Pink1 mRNA association upon insulin addition is required for PINK1 protein activation and its function as a ubiquitin kinase in the mitophagy pathway, thus placing PINK1 function under metabolic control. Induction of insulin resistance in vitro by the key genetic Alzheimer risk factor apolipoprotein E4 retains Pink1 mRNA at the mitochondria and prevents proper PINK1 activity, especially in neurites. Our results thus identify a metabolic switch controlling Pink1 mRNA localization and PINK1 activity via insulin and AMPK signalling in neurons and propose a mechanistic connection between insulin resistance and mitochondrial dysfunction., (© 2024. The Author(s).)

Nonreciprocal wave propagation arises in systems with broken time-reversal symmetry and is key to the functionality of devices, such as isolators or circulators, in microwave, photonic, and acoustic applications. In magnetic systems, collective wave excitations known as magnon quasiparticles have so far yielded moderate nonreciprocities, mainly observed by means of incoherent thermal magnon spectra, while their occurrence as coherent spin waves (magnon ensembles with identical phase) is yet to be demonstrated. Here, we report the direct observation of strongly nonreciprocal propagating coherent spin waves in a patterned element of a ferromagnetic bilayer stack with antiparallel magnetic orientations. We use time-resolved scanning transmission X-ray microscopy (TR-STXM) to directly image the layer-collective dynamics of spin waves with wavelengths ranging from 5 μm down to 100 nm emergent at frequencies between 500 MHz and 5 GHz. The experimentally observed nonreciprocity factor of these counter-propagating waves is greater than 10 with respect to both group velocities and specific wavelengths. Our experimental findings are supported by the results from an analytic theory, and their peculiarities are further discussed in terms of caustic spin-wave focusing.

Phase-shifting electron holography is an excellent method to reveal electron wave phase information with very high phase sensitivity over a large range of spatial frequencies. It circumvents the limiting trade-off between fringe spacing and visibility of standard off-axis holography. Previous implementations have been limited by the independent drift of biprism and sample. We demonstrate here an advanced drift correction scheme for the hologram series that exploits the presence of an interface of the TEM specimen to the vacuum area in the hologram. It allows to obtain reliable phase information up to 2π/452 at the 1 Å information limit of the Titan 80-300 kV environmental transmission electron microscope used, by applying a moderate voltage of 250 V to a single biprism for a fringe spacing of 1 Å. The obtained phase and amplitude information is validated at a thin Pt sample by use of multislice image simulation with the frozen lattice approximation and shows excellent agreement. The presented method is applicable in any TEM equipped with at least one electron biprism and thus enables achieving high resolution off-axis holography in various instruments including those for in-situ applications. A software implementation for the acquisition, calibration and reconstruction is provided., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Abstract: Background: After severe injury, optical measures of microvascular blood flow (MBF) decrease and do not normalize with resuscitation to normal blood pressure. These changes are associated with organ dysfunction, coagulopathy, and death. However, the pathophysiology is not well understood. Several possible pathways could also contribute to the development of trauma-induced coagulopathy (TIC). A small-animal model of trauma-related MBF derangement that persists after resuscitation and includes TIC would facilitate further study. Parametric contrast-enhanced ultrasound (CEUS) is particularly advantageous in this setting, because it noninvasively assesses MBF in large, deep vascular beds. We sought to develop such a model, measuring MBF with CEUS. Methods: Sixteen male Sprague-Dawley rats were anesthetized, ventilated, and cannulated. Rats were subjected to either no injury (sham group) or a standardized polytrauma and pressure-targeted arterial catheter hemorrhage with subsequent whole blood resuscitation (trauma group). At prespecified time points, CEUS measurements of uninjured quadriceps muscle, viscoelastic blood clot strength, and complete blood counts were taken. Results: After resuscitation, blood pressure normalized, but MBF decreased and remained low for the rest of the protocol. This was primarily driven by a decrease in blood volume with a relative sparing of blood velocity. Viscoelastic blood clot strength and platelet count also decreased and remained low throughout the protocol. Conclusion: We present a rat model of MBF derangement in uninjured skeletal muscle and coagulopathy after polytrauma that persists after resuscitation with whole blood to normal macrohemodynamics. Parametric CEUS analysis shows that this change is primarily due to microvascular obstruction. This platform can be used to develop a deeper understanding of this important process., (Copyright © 2023 by the Shock Society.)

We demonstrated resonance-based detection of magnetic nanoparticles employing novel designs based upon planar (on-chip) microresonators that may serve as alternatives to conventional magnetoresistive magnetic nanoparticle detectors. We detected 130 nm sized magnetic nanoparticle clusters immobilized on sensor surfaces after flowing through PDMS microfluidic channels molded using a 3D printed mold. Two detection schemes were investigated: (i) indirect detection incorporating ferromagnetic antidot nanostructures within microresonators, and (ii) direct detection of nanoparticles without an antidot lattice. Using scheme (i), magnetic nanoparticles noticeably downshifted the resonance fields of an antidot nanostructure by up to 207 G. In a similar antidot device in which nanoparticles were introduced via droplets rather than a microfluidic channel, the largest shift was only 44 G with a sensitivity of 7.57 G/ng. This indicated that introduction of the nanoparticles via microfluidics results in stronger responses from the ferromagnetic resonances. The results for both devices demonstrated that ferromagnetic antidot nanostructures incorporated within planar microresonators can detect nanoparticles captured from dispersions. Using detection scheme (ii), without the antidot array, we observed a strong resonance within the nanoparticles. The resonance's strength suggests that direct detection is more sensitive to magnetic nanoparticles than indirect detection using a nanostructure, in addition to being much simpler.

Free-ranging cats (Felis catus) are globally distributed invasive carnivores that markedly impact biodiversity. Here, to evaluate the potential threat of cats, we develop a comprehensive global assessment of species consumed by cats. We identify 2,084 species eaten by cats, of which 347 (16.65%) are of conservation concern. Islands contain threefold more species of conservation concern eaten by cats than continents do. Birds, reptiles, and mammals constitute ~90% of species consumed, with insects and amphibians being less frequent. Approximately 9% of known birds, 6% of known mammals, and 4% of known reptile species are identified in cat diets. 97% of species consumed are <5 kg in adult body mass, though much larger species are also eaten. The species accumulation curves are not asymptotic, indicating that our estimates are conservative. Our results demonstrate that cats are extreme generalist predators, which is critical for understanding their impact on ecological systems and developing management solutions., (© 2023. The Author(s).)

This study aimed to evaluate the efficacy of cryopreserved amniotic membrane (AM) grafts in chronic wound healing, including the mean percentage of wound closure per one AM application, and to determine whether the healing efficiency differs between AM grafts obtained from different placentas. A retrospective study analyzing inter-placental differences in healing capacity and mean wound closure after the application of 96 AM grafts prepared from nine placentas. Only the placentas from which the AM grafts were applied to patients suffering from long-lasting non-healing wounds successfully healed by AM treatment were included. The data from the rapidly progressing wound-closure phase (p-phase) were analyzed. The mean efficiency for each placenta, expressed as an average of wound area reduction (%) seven days after the AM application (baseline, 100%), was calculated from at least 10 applications. No statistical difference between the nine placentas' efficiency was found in the progressive phase of wound healing. The 7-day average wound reduction in particular placentas varied from 5.70 to 20.99% (median from 1.07 to 17.75) of the baseline. The mean percentage of wound surface reduction of all analyzed defects one week after the application of cryopreserved AM graft was 12.17 ± 20.12% (average ± SD). No significant difference in healing capacity was observed between the nine placentas. The data suggest that if there are intra- and inter-placental differences in AM sheets' healing efficacy, they are overridden by the actual health status of the subject or even the status of its individual wounds., (© 2023. The Author(s).)

Efficient generation and control of spin currents launched by terahertz (THz) radiation with subsequent ultrafast spin-to-charge conversion is the current challenge for the next generation of high-speed communication and data processing units. Here, we demonstrate that THz light can efficiently drive coherent angular momentum transfer in nanometer-thick ferromagnet/heavy-metal heterostructures. This process is non-resonant and does neither require external magnetic fields nor cryogenics. The efficiency of this process is more than one order of magnitude higher as compared to the recently observed THz-induced spin pumping in MnF 2 antiferromagnet. The coherently driven spin currents originate from the ultrafast spin Seebeck effect, caused by a THz-induced temperature imbalance in electronic and magnonic temperatures and fast relaxation of the electron-phonon system. Owing to the fact that the electron-phonon relaxation time is comparable with the period of a THz wave, the induced spin current results in THz second harmonic generation and THz optical rectification, providing a spintronic basis for THz frequency mixing and rectifying components., (© 2023. The Author(s).)

Transition-metal nitrides/nitrenes are highly promising reagents for catalytic nitrogen-atom-transfer reactivity. They are typically prepared in situ upon optically induced N 2 elimination from azido precursors. A full exploitation of their catalytic potential, however, requires in-depth knowledge of the primary photo-induced processes and the structural/electronic factors mediating the N 2 loss with birth of the terminal metal-nitrogen core. Using femtosecond infrared spectroscopy, we elucidate here the primary molecular-level mechanisms responsible for the formation of a unique platinum(II) nitrene with a triplet ground state from a closed-shell platinum(II) azide precursor. The spectroscopic data in combination with quantum-chemical calculations provide compelling evidence that product formation requires the initial occupation of a singlet excited state with an anionic azide diradical ligand that is bound to a low-spin d 8 -configured Pt II ion. Subsequent intersystem crossing generates the Pt-bound triplet azide diradical, which smoothly evolves into the triplet nitrene via N 2 loss in a near barrierless adiabatic dissociation. Our data highlight the importance of the productive, N 2 -releasing state possessing azide ππ* character as a design principle for accessing efficient N-atom-transfer catalysts., (© 2023 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

A novel setup for measuring magnetic fields of antiferromagnets (i.e., quadrupolar or higher-order magnetic fields) and generally weakly remanent magnetic materials is presented. The setup features a highly sensitive superconducting quantum interference device magnetometer with a magnetic field resolution of ∼ 10 fT and non-electric temperature control of the sample space for a temperature range of 1.5-65 K with a non-electric sample movement drive and optical position encoding. To minimize magnetic susceptibility effects, the setup components are degaussed and realized with plastic materials in sample proximity. Running the setup in magnetically shielded rooms allows for a well-defined ultra-low magnetic background field well below 150 nT in situ. The setup enables studies of inherently weak magnetic materials, which cannot be measured with high field susceptibility setups, optical methods, or neutron scattering techniques, giving new opportunities for the research on, e.g., spin-spiral multiferroics, skyrmion materials, and spin ices., (© 2023 Author(s). Published under an exclusive license by AIP Publishing.)

Background: The use of cementless femoral stems in patients undergoing primary total hip arthroplasty (THA) with Dorr C bone remains controversial for fear of fracture or subsidence. Purpose of this multicenter study was to compare clinical outcomes and complications of THA using a tapered femoral prosthesis in patients with Dorr C bone versus Dorr A/B bone., Methods: A total of 1,030 patients underwent primary THA with a tapered wedge femoral stem at a minimum one year follow up. Forty-eight patients with Dorr C bone (mean age 68.7 years) were compared with a matched cohort of patients with Dorr A/B bone (mean age 69.9 years). Mean follow-up was approximately 4 years in both cohorts. There were no differences in sex, age, body mass index (BMI), Harris Hip Score (HHS), complications, and radiographic outcomes including subsidence and aseptic loosening were evaluated., Results: Postoperative HHSs were a mean of 82 points in the Dorr C cohort compared to 84 points in the Door A/B cohort (P=0.2653). There was no significant difference in complication or revision rates for any reason (P=0.23). Mean subsidence for the Dorr C and Dorr A/B was 1.4 and 1.2 mm, respectively (P=0.5164), and there was no aseptic loosening of the femoral component found in either group., Conclusions: Current generation tapered wedge cementless femoral stems provide stable fixation for patients with Dorr C bone quality without increased complications with respect to fracture or subsidence and can be considered an alternative to cemented stems in patients with compromised bone quality., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://atm.amegroups.com/article/view/10.21037/atm.2019.08.124/coif). RD declares the following outside of the submitted work: Baltimore City Medical Society: board or committee member; Flexion Therapeutics: research support; Orthofix, Inc.: research support; Stryker: research support; tissue Gene: research support; United Orthopedics: research support. MAM declares the following outside of the submitted work: AAOS: board or committee member; American Association of Hip and Knee Surgeons: board or committee member; Cymedica: paid consultant; DJ Orthopaedics: paid consultant; research support; Flexion Therapeutics: paid consultant; Johnson & Johnson: paid consultant; research support; Journal of Arthroplasty: editorial or governing board; Journal of Knee Surgery: editorial or governing board; Knee Society: board or committee member; Medicus Works LLC: publishing royalties, financial or material support; Microport: IP royalties; National Institutes of Health (NIAMS & NICHD): research support; Ongoing Care Solutions: paid consultant; research support; Orthopedics: editorial or governing board; Orthosensor: paid consultant; research support; Pacira: paid consultant; Peerwell: paid consultant; stock or stock options; Performance Dynamics: paid consultant; Pfizer: paid consultant; Skye Biologics: paid consultant; Stryker: IP royalties; paid consultant; research support; Surgical Techniques International: editorial or governing board; TissueGene: paid consultant; research support; Up-to-Date: publishing royalties, financial or material support; USMI: stock or stock options; Wolters Kluwer Health-Lippincott Williams & Wilkins: publishing royalties, financial or material support. ALM declares the following outside of the submitted work: AAOS: board or committee member; Journal of Arthroplasty: editorial or governing board; Stryker: IP royalties; paid consultant; paid presenter or speaker; research support. The other authors have no conflicts of interest to declare., (2023 Annals of Translational Medicine. All rights reserved.)

Background: Wastewater surveillance provided early indication of COVID-19 in US municipalities. Residents of long-term care facilities (LTCFs) experienced disproportionate morbidity and mortality early in the COVID-19 pandemic. We implemented LTCF building-level wastewater surveillance for SARS-CoV-2 at 6 facilities in Kentucky to provide early warning of SARS-CoV-2 in populations considered vulnerable., Objective: This study aims to evaluate the performance of wastewater surveillance for SARS-CoV-2 at LTCFs in Kentucky., Methods: We conducted a mixed methods evaluation of wastewater surveillance following Centers for Disease Control and Prevention (CDC) guidelines for evaluating public health surveillance systems. Evaluation steps in the CDC guidelines were engaging stakeholders, describing the surveillance system, focusing the evaluation design, gathering credible evidence, and generating conclusions and recommendations. We purposively recruited stakeholders for semistructured interviews and undertook thematic content analysis of interview data. We integrated wastewater, clinical testing, and process data to characterize or calculate 7 surveillance system performance attributes (simplicity, flexibility, data quality, sensitivity and positive predictive value [PPV], timeliness, representativeness, and stability)., Results: We conducted 8 stakeholder interviews. The surveillance system collected wastewater samples (N=811) 2 to 4 times weekly at 6 LTCFs in Kentucky from March 2021 to February 2022. Synthesis of credible evidence indicated variable surveillance performance. Regarding simplicity, surveillance implementation required moderate human resource and technical capacity. Regarding flexibility, the system efficiently adjusted surveillance frequency and demonstrated the ability to detect additional pathogens of interest. Regarding data quality, software identified errors in wastewater sample metadata entry (110/3120, 3.53% of fields), technicians identified polymerase chain reaction data issues (140/7734, 1.81% of reactions), and staff entered all data corrections into a log. Regarding sensitivity and PPV, using routine LTCF SARS-CoV-2 clinical testing results as the gold standard, a wastewater SARS-CoV-2 signal of >0 RNA copies/mL was 30.6% (95% CI 24.4%-36.8%) sensitive and 79.7% (95% CI 76.4%-82.9%) specific for a positive clinical test at the LTCF. The PPV of the wastewater signal was 34.8% (95% CI 27.9%-41.7%) at >0 RNA copies/mL and increased to 75% (95% CI 60%-90%) at >250 copies/mL. Regarding timeliness, stakeholders received surveillance data 24 to 72 hours after sample collection, with delayed reporting because of the lack of weekend laboratory staff. Regarding representativeness, stakeholders identified challenges delineating the population contributing to LTCF wastewater because of visitors, unknown staff toileting habits, and the use of adult briefs by some residents preventing their waste from entering the sewer system. Regarding stability, the reoccurring cost to conduct 1 day of wastewater surveillance at 1 facility was approximately US $144.50, which included transportation, labor, and materials expenses., Conclusions: The LTCF wastewater surveillance system demonstrated mixed performance per CDC criteria. Stakeholders found surveillance feasible and expressed optimism regarding its potential while also recognizing challenges in interpreting and acting on surveillance data., (©James W Keck, Jess Lindner, Matthew Liversedge, Blazan Mijatovic, Cullen Olsson, William Strike, Anni Noble, Reuben Adatorwovor, Parker Lacy, Ted Smith, Scott M Berry. Originally published in JMIR Public Health and Surveillance (https://publichealth.jmir.org), 29.08.2023.)

Chronic thromboembolic pulmonary hypertension (CTEPH) is successfully treatable with pulmonary endarterectomy (PEA), balloon pulmonary angioplasty, and medical therapy. Registry to Evaluate Early and Long-Term Pulmonary Arterial Hypertension Disease Management risk score (RRS) is able to predict long-term outcome in inoperable patients or in patients with residual PH after surgery. We performed a post hoc analysis of RRS in patients who were enrolled in the CTREPH study (NCT01416636), a randomized, double-blind clinical trial comparing high-dose and low-dose subcutaneous (SC) treprostinil in patients with severe CTEPH that was classified by an interdisciplinary CTEPH team as nonoperable, or as persistent or recurrent pulmonary hypertension after PEA. Baseline mean RRS was similar in both treatment groups (8.7 in high-dose arm vs. 8.6 in low-dose arm), but mean RRS change from baseline to Week 24 was greater in the high-dose treprostinil group than in the low-dose treprostinil group (-0.88 vs. -0.17). The difference in RRS change from baseline to Week 24 between high dose versus low dose was statistically significant with mean difference of -0.70 (95% confidence interval: -1.36 to -0.05, p  = 0.0352), and was driven mainly by improvement of World Health Organization functional class and N-terminal pro-brain natriuretic peptide concentration. SC treprostinil therapy administered in standard dose had positive effect on the risk profile measured by RRS in patients with inoperable or persistent/recurrent severe CTEPH. Although our study was limited by the small sample size and post hoc nature, assessment of risk profile is of great importance to this particular patient population with very poor prognosis., Competing Interests: P. J. has received fees and grants from Actelion Pharmaceuticals Ltd., AOP Orphan, Bayer HealthCare, Merck Sharp & Dohme, and GlaxoSmithKline. He has served on advisory boards for Actelion Pharmaceuticals Ltd, Bayer HealthCare, and Merck Sharp & Dohme, outside of the Article. R. S.‐K. has relationships with drug companies including Actelion, AOP Orphan Pharmaceuticals, Bayer Schering Pharma, GlaxoSmithKline, and SciPharm Sàrl; is an investigator in trials involving these companies, relationships include consultancy service, and research grants. M. H. reports personal fees (lectures) from Acceleron, Actelion, AstraZeneca, Bayer Pharma AG, Berlin Chemie, GlaxoSmithKline, Janssen‐Cilag, Merck Sharp & Dohme, and Optimal Medical Therapies, nonfinancial support (travel) from Actelion, Janssen‐Cilag, Novartis and Optimal Medical Therapies, and is on an advisory board of Actelion, Glaxo‐SmithKline, Janssen‐Cilag and Merk Sharp & Dohme, outside of the Article. I. S. has relationships with Actelion, AOP Orphan Pharmaceuticals, Bayer Healthcare, Merck Sharp & Dohme, GlaxoSmithKline, and Pfizer, and is an investigator in trials involving these companies, relationships include consultancy service, research grants, and membership of scientific advisory boards, outside of the Article. I. M. L. has relationships with Actelion‐Janssen, AOP Orphan Pharmaceuticals, Bayer‐Schering, Daiichi Sankyo, Ferrer, SciPharm Sàrl and MSD, and is an investigator in trials involving these companies; has relationships including consultancy service, research grants, and scientific advisory boards. The remaining authors declare no conflict of interest., (© 2023 The Authors. Pulmonary Circulation published by John Wiley & Sons Ltd on behalf of Pulmonary Vascular Research Institute.)

To compare the therapeutic efficacy of cryopreserved amniotic membrane (AM) grafts and standard of care (SOC) in treating nonhealing wounds (NHW) through a prospective multicenter clinical trial, 42 patients (76% polymorbid) with 54 nonhealing wounds of various etiologies (mainly venous) and an average baseline size of 20 cm 2 were included. All patients were treated for at least 6 weeks in the center before they were involved in the study. In the SOC group, 29 patients (36 wounds) were treated. If the wound healed less than 20% of the baseline size after 6 weeks, the patient was transferred to the AM group (35 patients, 43 wounds). Weekly visits included an assessment of the patient's condition, photo documentation, wound debridement, and dressing. Quality of life and the pain degree were subjectively reported by patients. After SOC, 7 wounds were healed completely, 1 defect partially, and 28 defects remained unhealed. AM application led to the complete closure of 24 wounds, partial healing occurred in 10, and 9 remained unhealed. The degree of pain and the quality of life improved significantly in all patients after AM application. This study demonstrates the effectiveness of cryopreserved AM grafts in the healing of NHW of polymorbid patients and associated pain reduction.