Pulmonary hypertension worsens outcome in left heart disease. Stiffening of the pulmonary artery may drive this pathology by increasing right ventricular dysfunction and lung vascular remodeling. Here we show increased stiffness of pulmonary arteries from patients with left heart disease that correlates with impaired pulmonary hemodynamics. Extracellular matrix remodeling in the pulmonary arterial wall, manifested by dysregulated genes implicated in elastin degradation, precedes the onset of pulmonary hypertension. The resulting degradation of elastic fibers is paralleled by an accumulation of fibrillar collagens. Pentagalloyl glucose preserves arterial elastic fibers from elastolysis, reduces inflammation and collagen accumulation, improves pulmonary artery biomechanics, and normalizes right ventricular and pulmonary hemodynamics in a rat model of pulmonary hypertension due to left heart disease. Thus, targeting extracellular matrix remodeling may present a therapeutic approach for pulmonary hypertension due to left heart disease.

Background Parametric mapping sequences in cardiovascular magnetic resonance (CMR) allow for non-invasive myocardial tissue characterization. However quantitative myocardial mapping is still limited by the need for local reference values. Confounders, such as field strength, vendors and sequences, make intersite comparisons challenging. This exploratory study aims to assess whether multi-site studies that control confounding factors provide first insights whether parametric mapping values are within pre-defined tolerance ranges across scanners and sites. Methods A cohort of 20 healthy travelling volunteers was prospectively scanned at three sites with a 3 T scanner from the same vendor using the same scanning protocol and acquisition scheme. A Modified Look-Locker inversion recovery sequence (MOLLI) for T1 and a fast low-angle shot sequence (FLASH) for T2 were used. At one site a scan-rescan was performed to assess the intra-scanner reproducibility. All acquired T1- and T2-mappings were analyzed in a core laboratory using the same post-processing approach and software. Results After exclusion of one volunteer due to an accidentally diagnosed cardiac disease, T1- and T2-maps of 19 volunteers showed no significant differences between the 3 T sites (mean & PLUSMN; SD [95% confidence interval] for global T1 in ms: site I: 1207 & PLUSMN; 32 [1192-1222]; site II: 1207 & PLUSMN; 40 [1184-1225]; site III: 1219 & PLUSMN; 26 [1207-1232]; p = 0.067; for global T2 in ms: site I: 40 & PLUSMN; 2 [39-41]; site II: 40 & PLUSMN; 1 [39-41]; site III 39 & PLUSMN; 2 [39-41]; p = 0.543). Conclusion Parametric mapping results displayed initial hints at a sufficient similarity between sites when confounders, such as field strength, vendor diversity, acquisition schemes and post-processing analysis are harmonized. This finding needs to be confirmed in a powered clinical trial.Trial registration ISRCTN14627679 (retrospectively registered)

Background Disruption of glucocorticoid (GC) signaling in osteoblasts results in a marked attenuation of acute antibody-induced arthritis. The role of endogenous GCs in chronic inflammatory arthritis is however not fully understood. Here, we investigated the impact of endogenous GC signaling in osteoblasts on inflammation and bone integrity under chronic inflammatory arthritis by inactivating osteoblastic GC signaling in a long-term K/BxN serum transfer-induced induced arthritis (STIA) model. Methods Intracellular GC signaling in osteoblasts was disrupted by transgenic (tg) overexpression of 11beta-hydroxysteroid dehydrogenase type 2 (11ss-HSD2). Inflammatory arthritis was induced in 5-week-old male tg mice and their wild type (WT) littermates by intraperitoneal (i.p.) injection of K/BxN serum while controls (CTRLs) received phosphate-buffered saline (PBS). In a first cohort, K/BxN STIA was allowed to abate until the endpoint of 42 days (STIA). To mimic rheumatic flares, a second cohort was additionally injected on days 14 and 28 with K/BxN serum (STIA (boost)). Arthritis severity was assessed daily by clinical scoring and ankle size measurements. Ankle joints were assessed histopathologically. Systemic effects of inflammation on long bone metabolism were analyzed in proximal tibiae by micro-computed tomography (& mu;CT) and histomorphometry. Results Acute arthritis developed in both tg and WT mice (STIA and STIA (boost)) and peaked around day 8. While WT STIA and tg STIA mice showed a steady decline of inflammation until day 42, WT STIA (boost) and tg STIA (boost) mice exhibited an arthritic phenotype over a period of 42 days. Clinical arthritis severity did not differ significantly between WT and tg mice, neither in the STIA nor in the STIA (boost) cohorts. Correspondingly, histological indices of inflammation, cartilage damage, and bone erosion showed no significant difference between WT and tg mice on day 42. Histomorphometry revealed an increased bone turnover in tg CTRL and tg STIA (boost) compared to WT CTRL and WT STIA (boost) animals, respectively. Conclusions In contrast to the previously reported modulating effects of endogenous GC signaling in osteoblasts during acute K/BxN STIA, this effect seems to perish during the chronic inflammatory and resolution phase. These findings indicate that endogenous GC signaling in osteoblasts may mainly be relevant during acute and subacute inflammatory processes.

A growing body of literature suggests the important role of the thalamus in cognition and neurodegenerative diseases. This study aims to elucidate whether the preoperative thalamic volume is associated with preoperative cognitive impairment (preCI) and whether it is predictive for postoperative cognitive dysfunction at 3 months (POCD). We enrolled 301 patients aged 65 or older and without signs of dementia who were undergoing elective surgery. Magnetic resonance imaging was conducted prior to surgery. Freesurfer (version 5.3.) was used to automatically segment the thalamus volume. A neuropsychological test battery was administered before surgery and at a 3 month follow-up. It included the computerized tests Paired Associate Learning (PAL), Verbal Recognition Memory (VRM), Spatial Span Length (SSP), Simple Reaction Time (SRT), the pen-and-paper Trail-Making-Test (TMT) and the manual Grooved Pegboard Test (GPT). Using a reliable change index, preCI and POCD were defined as total Z-score > 1.96 (sum score over all tests) and/or Z-scores > 1.96 in >= 2 individual cognitive test parameters. For statistical analyses, multivariable logistic regression models were applied. Age, sex and intracranial volume were covariates in the models. Of 301 patients who received a presurgical neuropsychological testing and MRI, 34 (11.3%) had preCI. 89 patients (29.5%) were lost to follow-up. The remaining 212 patients received a follow-up cognitive test after 3 months, of whom 25 ( 8.3%) presented with POCD. Independently of age, sex and intracranial volume, neither preCI (OR per cm(3) increment 0.81 [95% CI 0.60-1.07] p = 0.14) nor POCD (OR 1.02 per cm(3) increment [95% CI 0.75-1.40] p = 0.87) were statistically significantly associated with patients' preoperative thalamus volume. In this cohort we could not show an association of presurgical thalamus volume with preCI or POCD.

Background In breast cancer patients body image (BI) is a crucial aspect of quality of life (QoL). This study examined the postoperative impact of different surgical approaches on long-term BI analyzing real-world data to guide pre- and postoperative patient care and preserve QoL. Methods EORTC QLQ-BR23 BI scores were collected electronically in 325 breast cancer patients within routine clinical care for a duration of 41.5 months (11/17/2016 - 4/30/2020) at predefined time points preoperatively and repeatedly up to two years after breast-conserving surgery (BCS) (n = 212), mastectomy alone (M) (n = 27) or mastectomy with immediate breast reconstruction (MIBR) (n = 86). Higher scores indicated better BI. A linear mixed regression model was used to analyze the impact of BCS, M and MIBR, as well as non-surgical therapies on BI at treatment initiation and over time. Results BI scores deteriorated by 5 points (95%-confidence interval (CI) -8.94 to -1.57, p & AP;0.005) immediately after BCS, by 7 points (95%-CI -12.13 to -1.80, p & AP;0.008) after MIBR and by 19 points (95%-CI -27.34 to -10.34, p < 0.001) after M. The change over time after BCS (+ 0.10 points per week, 95%-CI -0.17 to 0.38), MIBR (-0.07 points per week, 95%-CI -0.35 to 0.20) and M (+ 0.14 points per week, 95%-CI -0.19 to 0.48) were not statistically significant (each p > 0.05). At treatment initiation chemotherapy was associated with a 22-point decline (95%-CI -25.39 to -17.87, p < 0.001) in BI score, while radiotherapy was associated with a 5-point increase (95%-CI 1.74 to 9.02, p & AP;0.004). However, over time chemotherapy was associated with a score recovery (+ 0.28 points per week, 95%-CI 0.19 to 0.37, p < 0.001), whereas for radiotherapy a trend towards BI deterioration was observed (-0.11 points per week, 95%-CI -0.23 to 0.02, p & AP;0.101). Conclusions Breast cancer surgery negatively affects BI. BCS and MIBR presumably harm BI less than M in the early postoperative period. Our data suggests BI to be deteriorating in the long term after MIBR while improving after BCS or M. Radiotherapy seems to have an additional negative long-term impact on BI. These findings should be confirmed in further studies to enable evidence-based patient information as part of preoperative shared decision-making and postoperative patient care.

The secondary use of health data is associated with enormous opportunities to improve healthcare efficiency and the creation of novel data-driven value propositions. Information Systems research has recognized this potential and focuses on that topic from various perspectives, such as data-driven value creation through health analytics or data platform ecosystems to facilitate data sharing. However, with emerging regulatory initiatives, such as the European Health Data Space, rules for the secondary use are changing, with implications for researchers, innovators, and healthcare organizations. The outcomes of data value creation efforts are mainly dependent on available data, with the assurance of high data quality remaining a challenge. To discuss how value can be created from the secondary use of health data, what challenges may arise, and how they can be addressed, a workshop on this topic was organized in November 2023, in which 19 academic researchers and 18 practitioners from 27 organizations participated. This workshop report highlights the challenges of and potential solutions for the secondary use of health data. While current Information Systems research often focuses on utilizing Electronic Health Records, the field could be enriched by understanding the secondary use of health data on an individual, organizational, and societal level.

The altered posterior question-mark incision for decompressive hemicraniectomy (DHC) was proposed to reduce the risk of intraoperative injury of the superficial temporal artery (STA) and demonstrated a reduced rate of wound-healing disorders after cranioplasty. However, decompression size during DHC is essential and it remains unclear if the new incision type allows for an equally effective decompression. Therefore, this study evaluated the efficacy of the altered posterior question-mark incision for craniectomy size and decompression of the temporal base and assessed intraoperative complications compared to a modified standard reversed question-mark incision. The authors retrospectively identified 69 patients who underwent DHC from 2019 to 2022. Decompression and preservation of the STA was assessed on postoperative CT scans and CT or MR angiography. Forty-two patients underwent DHC with the standard reversed and 27 patients with the altered posterior question-mark incision. The distance of the margin of the craniectomy to the temporal base was 6.9 mm in the modified standard reversed and 7.2 mm in the altered posterior question-mark group (p = 0.77). There was no difference between the craniectomy sizes of 158.8 mm and 158.2 mm, respectively (p = 0.45), and there was no difference in the rate of accidental opening of the mastoid air cells. In both groups, no transverse/sigmoid sinus was injured. Twenty-four out of 42 patients in the modified standard and 22/27 patients in the altered posterior question-mark group had a postoperative angiography, and the STA was preserved in all cases in both groups. Twelve (29%) and 5 (19%) patients underwent revision due to wound-healing disorders after DHC, respectively (p = 0.34). There was no difference in duration of surgery. Thus, the altered posterior question-mark incision demonstrated technically equivalent and allows for an equally effective craniectomy size and decompression of the temporal base without increasing risks of intraoperative complications. Previously described reduction in wound-healing complications and cranioplasty failures needs to be confirmed in prospective studies to demonstrate the superiority of the altered posterior question-mark incision.

Nanoplastics (NPs) are emerging contaminants of concern that may interact with natural biotic stressors (such as parasites) to disrupt host-associated microbiomes, which play a crucial role in the health and ecological dynamics of aquatic animals. Here, we investigate the effects of polystyrene NP beads and parasite infection on the microbiome diversity and composition of the model plankton organism Daphnia magna. We exposed D. magna to two NP sizes (50 nm and 100 nm) at two concentrations (1 mg L−1 and 5 mg L−1), both with and without infection by the yeast parasite Metschnikowia bicuspidata and sequenced the microbiomes of gut and body tissues using 16S rRNA gene sequencing. High concentrations of 50 nm NPs significantly increased bacterial richness in both gut and body tissue, with shifts exceeding those induced by parasite infection. In the gut, the relative abundances of Burkholderiales and Chitinophagales decreased, while Caulobacterales, Rhizobiales, and Salinisphaerales increased. In body tissues, Chitinophagales declined, whereas Burkholderiales, Caulobacterales, Rhizobiales, and Salinisphaerales were enriched. NP size, concentration and interaction with infection, significantly influenced gut and body microbiome alpha diversity. Bray-Curtis dissimilarity analysis confirmed that 50 nm NPs drove distinct shifts in bacterial community composition, independent of parasite infection. Overall, NP-size and concentration had a stronger influence on the Daphnia microbiome than parasite infection. Given the critical roles of the Daphnia microbiome in nutritional support and stress tolerance, our findings highlight the potential ecological impacts of NPs on aquatic ecosystems.

Background The "UFO procedure" was initially developed as a surgical option to enlarge the aortic annulus in patients requiring valve replacement. This technique can be employed to treat extensive endocarditis located in the intervalvular fibrous body (IVFB). One of the indications for a "UFO procedure" is massive aortic and mitral valve calcification. It is a challenging surgical procedure with a high risk of intraoperative complications. Case summary We present a 76-year-old male patient with massive aortic and mitral valve calcification involving the left atrium, the left ventricle and the left ventricular outflow tract. Both valves exhibited severe stenosis and moderate to severe regurgitation. The left ventricle was hypertrophic and the left ventricular ejection fraction was > 55%. The patient was prediagnosed with persistent atrial fibrillation. The risk of death following heart surgery (EuroSCORE II) was calculated as 9.21%. We successfully performed a so-called "UFO procedure" including replacement of both valves without annular decalcification to avoid atrioventricular dehiscence. We enlarged the IVFB and replaced the non-coronary sinus of Valsalva with doubled bovine pericardium. The left ventricular outflow tract was decalcified. The patient was transferred to a local hospital on the 13th postoperative day. Conclusion Successful surgical treatment to this extent was demonstrated for the first time. Due to the high perioperative mortality, the surgical treatment of patients with this constellation would be refused in most cases. In our patient, the preoperative imaging showed extreme calcification of both valves and the surrounding myocardium. Excellent preoperative planning and a highly experienced surgical team is necessary.

Objectives SARS-CoV-2 virus infection can lead to acute respiratory distress syndrome (ARDS), which can be complicated by severe muscle wasting. Until now, data on muscle loss of critically ill COVID-19 patients are limited, while computed tomography (CT) scans for clinical follow-up are available. We sought to investigate the parameters of muscle wasting in these patients by being the first to test the clinical application of body composition analysis (BCA) as an intermittent monitoring tool. Materials BCA was conducted on 54 patients, with a minimum of three measurements taken during hospitalization, totaling 239 assessments. Changes in psoas- (PMA) and total abdominal muscle area (TAMA) were assessed by linear mixed model analysis. PMA was calculated as relative muscle loss per day for the entire monitoring period, as well as for the interval between each consecutive scan. Cox regression was applied to analyze associations with survival. Receiver operating characteristic (ROC) analysis and Youden index were used to define a decay cut-off. Results Intermittent BCA revealed significantly higher long-term PMA loss rates of 2.62% (vs. 1.16%, p < 0.001) and maximum muscle decay of 5.48% (vs. 3.66%, p = 0.039) per day in non-survivors. The first available decay rate did not significantly differ between survival groups but showed significant associations with survival in Cox regression (p = 0.011). In ROC analysis, PMA loss averaged over the stay had the greatest discriminatory power (AUC = 0.777) for survival. A long-term PMA decline per day of 1.84% was defined as a threshold; muscle loss beyond this cut-off proved to be a significant BCA-derived predictor of mortality. Conclusion Muscle wasting in critically ill COVID-19 patients is severe and correlates with survival. Intermittent BCA derived from clinically indicated CT scans proved to be a valuable monitoring tool, which allows identification of individuals at risk for adverse outcomes and has great potential to support critical care decision-making.

Hybrid materials consisting of molybdenum disulfide (MoS2) and graphitic-like carbon have great potential for practical application as anodes in high-performance sodium-ion batteries. In this work, to reveal the effect of carbon coating on the interaction of sodium with the MoS2 layers located vertically relative to the substrate, model experiments were carried out using synchrotron-radiation-induced X-ray photoelectron spectroscopy (XPS). Sodium vapor obtained by heating a sodium source was simultaneously deposited in vacuum on the surfaces of MoS2, pyrolytic carbon, and a hybrid sample obtained by transferring a pyrolytic carbon film onto the MoS2 film. According to XPS data, sodium easily penetrates into the space between the vertical layers of the uncoated film, and its interaction with MoS2 leads to the transformation of the original hexagonal structure into a distorted tetragonal one. Under the experimental conditions, sodium is unable to diffuse through the carbon film consisting of horizontally oriented graphene domains and is almost completely removed by annealing the sample at 773 K in ultrahigh vacuum. The presence of the underlying MoS2 film facilitates the diffusion of sodium through the graphitic coating, but not all of the deposited sodium reaches MoS2. As a result, the sodium-induced rearrangement of the carbon-coated MoS2 is less than that of the free MoS2 film, and annealing of the sodiated sample restores its structure. The obtained results demonstrate the important role of the graphitic coating in the development of viable MoS2-based electrodes for energy storage systems.

Background Neurological disorders account for a large and increasing proportion of the global burden of disease. Therefore, it is important to strengthen the management of neurologic care, particularly in rural areas. The use of tele-neurology in primary care in rural areas is internationally considered to have the potential to increase access to health care services and improve the quality of care in these underserved areas. NeTKoH aims to address the existing knowledge gap regarding the effects of a tele-neurologic intervention in primary care under real-world conditions in a rural area in Germany. Methods NeTKoH is a cluster-randomized controlled trial with a stepped-wedge design involving 33 outpatient general practitioner's (GP) offices (clusters) in a rural area in Northeast Germany. During 11 predetermined steps, all clusters are randomized before they cross over into groups from the control to the intervention arm. The targeted sample size is 1,089 patients with neurologic symptoms that are continuously being recruited. In the intervention arm, tele-neurologic consultations will be provided via a face-to-face video conferencing system with a neurologic expert at a university hospital. The control arm will receive usual care. The primary outcome is the proportion of neurologic problems being solved at the GP's office. Secondary outcomes will comprise hospital stays and days, time until neurologic specialist appointments and diagnostics, patients' health status and quality of life, outpatient and inpatient referrals.A concurrent observational study, together with a process, implementation, and health economic evaluation, will also be conducted. Discussion Using a stepped-wedge cluster design in a real-life situation can help with logistic challenges and enhance the motivation of the participating GPs, as all, at some point, will be in the intervention phase. With the additional implementation evaluation pertaining to external validity, an observational study, and a health economic evaluation, NeTKoH will be able to provide an extensive evaluation for health policy decision-makers regarding the uptake into standard care.

Background Hypertension is a highly relevant public health challenge. Digital interventions may support improving adherence to anti-hypertensive medications and alter health behavior. Therefore, this protocol describes a study that aims to assess the effectiveness of mHealth and educational support through peer counseling (Ed-counselling) to control blood pressure in hypertensive patients when compared to standard care.Methods We chose a double-blinded pragmatic randomized-controlled with factorial design for this investigation. The trial is going to recruit 1648 hypertensive patients with coronary artery disease at the age of 21 to 70 years. All participants will already be on anti-hypertensive medication and own a smartphone. They will be randomized into four groups with each having 412 participants. The first group will only receive standard care; while the second group, in addition to standard care, will receive monthly Ed-counselling (educational booklets with animated infographics and peer counseling); the third group will receive daily written and voice reminders and an education-led video once weekly together with standard care; while the fourth one gets both interventions given to second and third groups respectively. All groups will be followed-up for 1 year (0, 6, and 12 months). The primary outcome will be the change in systolic blood pressure while secondary outcomes include health-related quality of life and changes in medication adherence. For measuring changes in systolic blood pressure (SBP) and adherence scores difference at 0, 6, and 12 months between and within the group, parametric (ANOVA/repeated measure ANOVA) and non-parametric tests (Kruskal-Wallis test/Friedman test) will be used. By using the general estimating equation (GEE) with negative binomial regression, at 12 months, the covariates affecting primary and secondary outcomes will be determined and controlled. The analysis will be intention-to-treat. All the outcomes will be analyzed at 0, 6, and 12 months; however, the final analysis will be at 12 months from baseline.Discussion Besides adding up to existing evidence in the literature on the subject, our designed modules using mHealth technology can help in reducing hypertension-related morbidity and mortality in developing countries.

A novel guanidine-based NNN pincer cobalt(II) complex, Co1, was investigated for dioxygen (O2) activation. Upon reaction with O2 at room temperature, a μ-hydroxo-bridged CoII–OH-CoII complex (Co2) was isolated as the final oxygenated product. In-depth spectroscopic investigation revealed that the formation of Co2 occurs via an intermediate superoxide species (Co1–O2•–), by taking advantage of the redox non-innocence behavior of the ligand (3,3′-(pyridine-2,6-diyl)bis(1-methyl-N-phenylimidazolidin-2-imine)), and therefore keeping the cobalt oxidation state unchanged at +2. Interestingly, the zinc analogue, Zn1, was also shown to yield a similar μ-hydroxo-bridged ZnII–OH-ZnII complex (Zn2) as the final product under the same aerobic conditions, providing further confirmation of the ligand influence on metal oxidation and spin states during O2 activation. Further reactivity studies demonstrated that Co1–O2•– reacts with nitric oxide (•NO) to form a CoII-peroxynitrite (Co1–ONO2–) intermediate, which performs an unprecedented intramolecular nitration of the phenyl moieties of the ligand at the para position.

Russian aggression against Ukraine shook the ‚balance of power’ in international relations and demonstrated a disruption in the ‘offense-defense balance’ worldwide. This paper advances Foreign Policy Analysis and International Relations by showcasing a new study of Russian foreign policy. This original research offers a unique study on congruency in analytical concepts of Eurasian regionalism and Neo-Eurasianism based on Arendt’s ideological conceptualization. The constructivist approach to Russian ideology and state identity broadens the scientific scope and enriches understanding of the ideational reasons for Russia’s invasion of Ukraine. The deeper insight into the regional cooperation in ‘Eurasia’ through Russian-chaired organizations indicates coherence between Eurasian regionalism and the postulates of Neo-Eurasianism. On this, Russia’s failure to lure Ukraine into regional cooperation with ‘soft’ power resulted in ‘hard’ militarism. The analysis of the original texts of Russian ideologist Dugin demonstrates that the reshaping of the world order starts with Ukraine, but it does not end.

Extreme temperature changes from one day to another that are associated with either warming or cooling can have a significant impact on health, the environment, and society. Previous studies have shown that such day-to-day temperature (DTDT) changes are typically more pronounced in the extratropics than the tropics. However, the underlying physical processes and the relationship between extreme events and the large-scale atmospheric circulation remain poorly understood. Here, these processes are investigated for different locations around the globe based on ERA5 reanalysis data and Lagrangian backward-trajectory calculations. We show that extreme DTDT changes in the extratropics are generally associated with changes in air mass transport, particularly shifts from warmer to colder air parcels or vice versa that are linked to regionally specific synoptic-scale circulation anomalies (ridge or trough patterns). These dominant effects of advection are modulated by changes in adiabatic and diabatic processes in the transported air parcels, which either amplify or dampen DTDT decreases (cooling events) and increases (warming events) depending on the region and season. In contrast, extreme DTDT changes during December–February in the tropics are controlled by local processes rather than changes in advection. For instance, the most significant DTDT decreases are associated with a shift from less cloudy to more cloudy conditions, highlighting the crucial role of solar radiative heating. The mechanistic insights into the extreme DTDT changes obtained in this study can help improve the prediction of such events and anticipate future changes in their occurrence frequency and intensity, which will be investigated in part 2 of this study.

Porcine cytomegalovirus/porcine roseolovirus (PCMV/PRV), a porcine herpesvirus, has been shown to significantly reduce the survival time of porcine xenotransplants in non-human primates. The virus was detected in all the examined organs of baboons transplanted with PCMV/PRV-positive organs and it was also transmitted to the first human recipient of a pig heart, contributing to the patient’s death. PCMV/PRV induces consumptive coagulopathy and thrombocytopenia in xenotransplant recipients. Initial studies in baboons revealed that the virus triggered increased release of tumor necrosis factor α (TNFα) and interleukin 6 (IL-6), along with elevated levels of tissue plasminogen activator (tPA) and plasminogen activator inhibitor 1 (PAI-1) complexes. Since there is no evidence that PCMV/PRV infects primate cells, including human cells, the virus appears to directly interact with immune and endothelial cells, disrupting cytokine signaling and coagulation pathways. The highest viral load was detected in the explanted pig heart, suggesting active replication at this site. Additionally, cells expressing PCMV/PRV proteins were identified in all the examined baboon organs, where pig cells were also found. Since PCMV/PRV affects only xenotransplant recipients and not healthy humans, this condition should be classified as a xenozoonosis. Interestingly, antibodies against human herpesvirus 6 (HHV-6) cross-react with PCMV/PRV and may contribute to protection against infection in humans. Further research is needed to uncover the molecular mechanisms underlying this xenozoonotic disease.

Porcine lymphotropic herpesviruses -1, -2, and -3 (PLHV-1, PLHV-2, and PLHV-3) are gammaherpesviruses that are widespread in pigs. These viruses are closely related to the human pathogens Epstein–Barr virus (EBV) and Kaposi sarcoma-associated herpesvirus (KSHV), both of which are known to cause severe diseases in humans. To date, however, no definitive association has been established between PLHVs and any disease in pigs. With the growing interest in xenotransplantation as a means to address the shortage of human organs for transplantation, the safety of using pig-derived cells, tissues, and organs is under intense investigation. In preclinical trials involving pig-to-nonhuman primate xenotransplantation, another porcine herpesvirus—porcine cytomegalovirus, a porcine roseolovirus (PCMV/PRV)—was shown to be transmissible and significantly reduced the survival time of the xenotransplants. In the present study, we examined donor pigs and their respective baboon recipients, all of which were part of preclinical pig heart xenotransplantation studies, for the presence of PLHV. PLHV-1, PLHV-2, and PLHV-3 were detected in nearly all donor pigs; however, no evidence of PLHV transmission to the baboon recipients was observed.

The open reading frame 3a (ORF3a) is a protein important to the pathogenicity of SARS-CoV-2. The cytoplasmic domain of ORF3a has three canonical tyrosine-based sorting signals (160YNSV163, 211YYQL213, and 233YNKI236), and a previous study has indicated that mutation of the 160YNSV163 motif abrogated plasma membrane expression and inhibited ORF3a-induced apoptosis. Here, we have systematically removed all three tyrosine-based motifs and assessed the importance of each motif or combination of motifs in trafficking to the cell surface. Our results indicate that the 160YNSV163 motif alone was insufficient for ORF3a cell-surface trafficking, while the 211YYQL213 motif was the most important. Additionally, an ORF3a with all three YxxΦ motifs disrupted (ORF3a-[ΔYxxΦ]) was not transported to the cell surface, and LysoIP studies indicate that ORF3a but not ORF3a-[ΔYxxΦ] was present in late endosome/lysosome fractions. A growth-curve analysis of different SARS-CoV-2 viruses expressing the different mutant ORF3a proteins revealed no significant differences in virus replication. Finally, the inoculation of K18hACE-2 mice indicated that the SARS-CoV-2 lacking the three YxxΦ motifs was less pathogenic than the unmodified SARS-CoV-2. These results indicate that the tyrosine motifs of ORF3a contribute to cell-surface expression and SARS-CoV-2 pathogenesis.