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'Flavescence dorée' (FD) is a quarantine grapevine disease associated with FD phytoplasmas (FDp). In Switzerland, FD was first reported in 2004 in the southern part of the Canton Ticino, from where, within a few years, it spread to nearly the entire vineyard area of the Southern Alps, despite the mandatory control measures. The aim of this study was to assess the possible role of gone‐wild grapevines (GWGVs) in forests resulting from early abandoned vineyards as a habitat for the main FDp vector, Scaphoideus titanus, and for the best alternative vector candidate, Orientus ishidae, as well as an FDp inoculum reservoir in Southern Switzerland. Leaf samples were collected in 20 plots and tested for the presence of FDp. Moreover, fifteen of these plots were monitored with yellow sticky traps to determine the presence and infection status of insect vectors. Finally, a hatching experiment under controlled conditions was conducted to investigate the possible oviposition activity by FDp vectors, using wood collected from GWGVs present in 11 forest sites in the surroundings of cultivated vineyards. GWGVs in forests were confirmed to act as an FDp reservoir and to represent a suitable habitat for FDp vectors (in particular, for S. titanus). Abandoned vineyards should be included in the FD management strategy and their systematic rogueing should be applied as soon as possible and in particular before the transition from vineyard to forest occurs, in order to avoid the survival of S. titanus and the establishment of inoculum in uncontrolled landscape compartments. [ABSTRACT FROM AUTHOR]

We previously demonstrated that lung subjected to a mild thermal preconditioning (TP) of 41.5°C during ex vivo lung perfusion (EVLP) had improved function and decreased inflammatory and cell death markers. How TP affects the populations of immune cells in the lung airways and the perfusion solution after EVLP is the purpose of this study. Male rats were randomly assigned into 2 groups: controls (Ctl, n=7) or TP (n=9). Lungs were kept in situ for 1h at room temperature and flushed with cold Perfadex. The heart-lung block was harvested, kept at 4°C for 1h and mounted on the EVLP system. In the Ctl group, the temperature of the perfusion solution was kept at 37°C. In the TP group, a heat shock was done after 1h of EVLP by heating the perfusion solution to 41.5°C for 30 minutes. At the end of EVLP, we sampled and centrifuged the bronchoalveolar lavage fluid (BALF) and the perfusion solution to collect cells. The BALF and perfusion solution contents of B lymphocytes, CD4+ and CD8+ T lymphocytes as well as contents in macrophages, monocytes and granulocytes were analyzed by flow cytometry and expressed as the percentages of live cells. We did not observe significant differences in total live cell counts in the perfusate and the BALF between the Ctl and TP groups. In the BALF, the proportions of immune cells were unchanged by TP as compared to Ctl. In the perfusate, we observed a significant reduction in T cells populations in TP group as compared to Ctl. CD4+ T cells were 40.5 ± 8.7% of viable cells in Ctl group and 26.9±4.5% (p=0.001) in the TP group. CD8+ were 25.8 ± 3.8% in Ctl and 19.6 ± 3.8% in TP (p=0.005). B lymphocytes were not changed (15.6 ± 5.2% in Ctl vs 18.8 ± 6.2% in TP). The proportion of myeloid cells were increased in TP with 14.0 ± 5.1% in Ctl and 23.7 ± 5.0% in TP (p=0.002). Myeloid cells were predominantly monocytes (13.3 ± 4.9% in Ctl and 21.5± 4.9% in TP (p=0.003)) and neutrophils (0.4 ± 0.3% in Ctl and 1.7± 1.0% in TP (p=0.005)). TP during EVLP reduces T lymphocytes and augments monocytes and neutrophils proportions in the perfusion solution. Whether this results from changes in endothelium adhesion capability or permeability in responses to TP and/or immune cell type specific responses to TP will need more investigations. This underlines the profound effects of TP on immune responses and opens an exciting field of research. [ABSTRACT FROM AUTHOR]

The heat shock response (HSR) is a highly conserved adaptative response to thermic stress, related to the induced expression of heat shock proteins (HSPs) which promote significant cytoprotection. We hypothesized that thermal preconditioning (TP) with a well-calibrated heat stress (HS) applied during EVLP elicits a HSR and improves lung graft function by interfering with immune inflammation and oxidative stress pathways. Donor lungs (n=12) underwent extended hypothermic preservation of 16h. Lungs were assigned into two groups: 1) Control (n=6): 4h EVLP at 37°C; and 2) TP, (n=6): normothermic EVLP (37°C, 2h), followed by hyperthermic EVLP (42°, 30min), and normothermic EVLP (37°C, 1.5h). Ex-vivo lung function was assessed by measuring airway pressure, lung compliance and pulmonary vascular resistance. We determined expression of HSPs in tissue, and lung damage and oxidative stress biomarkers in tissue, BAL and perfusate at multiple time points. Lungs undergoing thermal preconditioning (TP group) showed significantly lower pulmonary arterial pressure and vascular resistance, increased dynamic and static pulmonary compliance and reduced airway pressure with no changes in oxygenation capacity as compared to lungs in the control group. At the end of EVLP, lungs in TP group had significant increase of lung mRNA of Hspa1a, Hsc70 and Hmox-1, reduction of nitrotyrosine (3-NT) in lung tissue, and reduction of LDH, CK18-M30 and proteins in BAL fluid. TP during ELVP decreased the expression of TNFα, IL6/IL10, MPO and IL-8 in BAL consistent with inflammasome inhibition. Transient heat stress during EVLP induces HSR and reduces nitroxidative stress, cellular injury and edema, and prevents functional deterioration of pig lungs after extended cold ischemia. These data indicate the therapeutic potential of thermal preconditioning as a way to recondition damaged lungs and to improve current preservation strategies. [ABSTRACT FROM AUTHOR]

The phloem-restricted phytopathogenic bacterium Candidatus Arsenophonus phytopathogenicus (Ap) causes the “syndrome basses richesses” (SBR), which has recently emerged as a major burden for sugar beets in several countries of Western Europe. Here, we report the first identification of Ap and its planthopper vector Pentastiridius leporinus in potato fields in Switzerland in 2023. The bacterium was detected in potato plants and tubers exhibiting phytoplasma-like symptoms and collected from cantons currently experiencing SBR outbreaks. Although our analyses indicate the absence of the Stolbur phytoplasma, the pathogenicity of Ap in potato remains unclear, a fortiori in the context of the poor phytosanitary status of crops in Switzerland in 2023. In infected tubers, we show that Ap can be detected after dormancy from the stem end to the emerging sprouts with decreasing titers. Importantly, Ap might induce threadlike sprouts and the browning of the flesh upon frying, raising strong concerns for varieties marketed for chips production. Altogether, our results align with recent studies performed in Germany, highlighting the host shift of Ap and its vector from sugar beet to potato crops. Our data should raise awareness in other European countries where the presence of the bacterium has been recently described. [ABSTRACT FROM AUTHOR]

Myocardial inflammation following myocardial infarction (MI) is crucial for proper myocardial healing, yet, dysregulated inflammation may promote adverse ventricular remodeling and heart failure. IL-1 signaling contributes to these processes, as shown by dampened inflammation by inhibition of IL-1β or the IL-1 receptor. In contrast, the potential role of IL-1α in these mechanisms has received much less attention. Previously described as a myocardial-derived alarmin, IL-1α may also act as a systemically released inflammatory cytokine. We therefore investigated the effect of IL-1α deficiency on post-MI inflammation and ventricular remodeling in a murine model of permanent coronary occlusion. In the first week post-MI, global IL-1α deficiency (IL-1α KO mice) led to decreased myocardial expression of IL-6, MCP-1, VCAM-1, hypertrophic and pro-fibrotic genes, and reduced infiltration with inflammatory monocytes. These early changes were associated with an attenuation of delayed left ventricle (LV) remodeling and systolic dysfunction after extensive MI. In contrast to systemic Il1a-KO, conditional cardiomyocyte deletion of Il1a (CmIl1a-KO) did not reduce delayed LV remodeling and systolic dysfunction. In conclusion, systemic Il1a-KO, but not Cml1a-KO, protects against adverse cardiac remodeling after MI due to permanent coronary occlusion. Hence, anti-IL-1α therapies could be useful to attenuate the detrimental consequences of post-MI myocardial inflammation. [ABSTRACT FROM AUTHOR]

Simple Summary: Annual rings of the susceptible grapevine cultivar 'Chardonnay' were measured and used in order to analyse the impact of the Flavescence dorée (FD) infection on the growth in diameter and the anatomical structure of grapevine trunks. Grapevines are susceptible to water shortage and reduce their growth in diameter in the case of summer drought. However, in the case of the expression of FD symptoms, the ring width reductions are extreme and supersede the drought-induced effects. In addition, in coincidence of the FD symptomatic expression, the anatomy of the phloem tissue of infected grapevines appears heavily disarranged. Moreover, sometimes also the formation of the woody ring is incomplete (early wood only). In conclusion, even though the FD phytoplasma does not inhabit and replicate inside the xylem tissue, our results confirm existing indirect inhibiting effects on the ring growth and the xylem tissue formation in FDp-infected grapevines. Flavescence dorée (FD) is a grapevine disease caused by 'Candidatus Phytoplasma vitis' (FDp), which is epidemically transmitted by the Nearctic leafhopper Scaphoideus titanus. In this study, we applied dendrochronological techniques to analyse the response to FDp infections in terms of wood ring widths and anatomical structures of the xylem and phloem tissues of the trunk of the susceptible grapevine cultivar 'Chardonnay.' As a rule, grapevines are susceptible to water shortage and reduce their growth in diameter in case of summer drought. In the season of the external expression of FD symptoms, however, the ring width reductions are extreme and supersede any drought-induced effects. In addition, the anatomy of the phloem tissue in the year of the FD symptom expression appears heavily disarranged. Moreover, in the most suffering individuals, the xylem formation remains incomplete and mostly limited to the early wood tissue. In conclusion, even though the FD phytoplasma does not inhabit and replicate inside the xylem tissue, our results confirm existing indirect inhibiting effects on the ring growth and the xylem tissue formation in FDp-infected grapevines. [ABSTRACT FROM AUTHOR]

Lung transplantation (LTx) is associated with sterile inflammation, possibly related to the release of damage associated molecular patterns (DAMPs) by injured allograft cells. We have measured cellular damage and the release of DAMPs and cytokines in an experimental model of LTx after cold or warm ischemia and examined the effect of pretreatment with ex-vivo lung perfusion (EVLP). Rat lungs were exposed to cold ischemia alone (CI group) or with 3h EVLP (CI-E group), warm ischemia alone (WI group) or with 3 hour EVLP (WI-E group), followed by LTx (2 hour). Bronchoalveolar lavage (BAL) was performed before (right lung) or after (left lung) LTx to measure LDH (marker of cellular injury), the DAMPs HMGB1, IL-33, HSP-70 and S100A8, and the cytokines IL-1β, IL-6, TNFα, and CXCL-1. Graft oxygenation capacity and static compliance after LTx were also determined. Compared to CI, WI displayed cellular damage and inflammation without any increase of DAMPs after ischemia alone, but with a significant increase of HMGB1 and functional impairment after LTx. EVLP promoted significant inflammation in both cold (CI-E) and warm (WI-E) groups, which was not associated with cell death or DAMP release at the end of EVLP, but with the release of S100A8 after LTx. EVLP reduced graft damage and dysfunction in warm ischemic, but not cold ischemic, lungs. The pathomechanisms of sterile lung inflammation during LTx are significantly dependent on the conditions. The release of HMGB1 (in the absence of EVLP) and S100A8 (following EVLP) may be important factors in the pathogenesis of LTx. [ABSTRACT FROM AUTHOR]

In vitro metabolic stability assays using rainbow trout (Oncorhynchus mykiss) isolated hepatocytes (RT‐HEP) or hepatic S9 fractions (RT‐S9) were introduced to provide biotransformation rate data for the assessment of chemical bioaccumulation in fish. The present study explored the suitability of the RT‐HEP and RT‐S9 assays for difficult test chemicals, and the in vitro–based predictions were compared to in silico–based predictions and in vivo–measured bioconcentration factors (BCFs). The results show that volatile or reactive chemicals can be tested with minor modifications of the in vitro protocols. For hydrophobic chemicals, a passive dosing technique was developed. Finally, a design‐of‐experiment approach was used to identify optimal in vitro assay conditions. The modified assay protocols were applied to 10 fragrances with diverse physicochemical properties. The in vitro intrinsic clearance rates were higher in the S9 than in the hepatocyte assay, but the in vitro–in vivo (IVIV) predictions were comparable between the 2 assays. The IVIV predictions classified the test chemicals as nonbioaccumulative (BCF < 2000), which was in agreement with the in vivo data but in contrast to the in silico–based predictions. The findings from the present study provide strong evidence that the RT‐HEP and RT‐S9 assays can provide reliable estimates of in vivo biotransformation rates for test chemicals with difficult physicochemical properties. Environ Toxicol Chem 2020;39:2396–2408. © 2020 SETAC [ABSTRACT FROM AUTHOR]

Potato blackleg is a seedborne disease that can cause significant economic losses for growers. Disease development depends mainly on two drivers, namely seed inoculum and local climatic conditions. To better establish the relationship between these two drivers, blackleg development was monitored in Swiss field trials at multiple locations from 2010 to 2013 involving three sets of naturally infected seed lots planted in each of three locations. The seed lot itself was thereby the most important factor explaining differences in disease development, rather than environmental factors. In a further on‐farm project conducted at various locations in Switzerland and southern Germany from 2013 to 2015, the implementation of a seed‐testing procedure was investigated. A total of 177 seed lots were tested for natural latent infection with soft rot Pectobacteriaceae and the corresponding blackleg incidence was tracked in 242 fields. The reliability of the relationship between latent infection and field incidence was found to be strongly linked to the bacterial species. Dickeya spp. field infection could be predicted with an acceptable reliability, whereas Pectobacterium carotovorum subsp. brasiliense, even when detected as latent tuber infection, was not consistently expressed as visual blackleg. Moreover, commonly found mixed latent infections with several bacterial species made it even harder to predict which bacteria would cause blackleg symptoms. Finally, variability in the reliability of seed testing may also be explained by differences in local farming practices. These trials over several years with naturally infected potato seed highlight the usefulness and limits of seed testing to manage blackleg. [ABSTRACT FROM AUTHOR]

The mineralocorticoid hormone aldosterone plays a crucial role in the control of Na+ and K+ balance, blood volume, and arterial blood pressure, by acting in the aldosterone‐sensitive distal nephron (ASDN) and stimulating a complex transcriptional, translational, and cellular program. Because the complexity of the aldosterone response is still not fully appreciated, we aimed at identifying new elements in this pathway. Here, we demonstrate that the expression of the proto‐oncogene PIM3 (Proviral Integration Site of Moloney Murine Leukemia Virus 3), a serine/threonine kinase belonging to the calcium/calmodulin‐regulated group of kinases, is stimulated by aldosterone in vitro (mCCDcl1 cells), ex vivo (mouse kidney slices), and in vivo in mice. Characterizing a germline Pim3−/− mouse model, we found that these mice have an upregulated Renin‐Angiotensin‐Aldosterone System (RAAS), with high circulating aldosterone and plasma renin activity levels on both standard or Na+‐deficient diet. Surprisingly, we did not observe any obvious salt‐losing phenotype in Pim3 KO mice as shown by normal blood pressure, plasma and urinary electrolytes, as well as unchanged expression levels of the major Na+ transport proteins. These observations suggest that the potential effects of the loss of the Pim3 gene are physiologically compensated. Indeed, the 2 other family members of the PIM kinase family, PIM1 and PIM2 are upregulated in the kidney of Pim3−/− mice, and may therefore be involved in such compensation. In conclusion, our data demonstrate that the PIM3 kinase is a novel aldosterone‐induced protein, but its precise role in aldosterone‐dependent renal homeostasis remains to be determined. [ABSTRACT FROM AUTHOR]

The E3 ubiquitin ligase NEDD4-2 (encoded by the Nedd4L gene) regulates the amiloride-sensitive epithelial Na+ channel (ENaC/SCNN1) to mediate Na+ homeostasis. Mutations in the human ß/?ENaC subunits that block NEDD4-2 binding or constitutive ablation of exons 6-8 of Nedd4L in mice both result in salt-sensitive hypertension and elevated ENaC activity (Liddle syndrome). To determine the role of renal tubular NEDD4-2 in adult mice, we generated tetracycline-inducible, nephron-specific Nedd4L KO mice. Under standard and high- Na+ diets, conditional KO mice displayed decreased plasma aldosterone but normal Na+/K+ balance. Under a high-Na+ diet, KO mice exhibited hypercalciuria and increased blood pressure, which were reversed by thiazide treatment. Protein expression of ßENaC, ?ENaC, the renal outer medullary K+ channel (ROMK), and total and phosphorylated thiazide-sensitive Na+Cl- cotransporter (NCC) levels were increased in KO kidneys. Unexpectedly, Scnn1a mRNA, which encodes the aENaC subunit, was reduced and proteolytic cleavage of aENaC decreased. Taken together, these results demonstrate that loss of NEDD4-2 in adult renal tubules causes a new form of mild, salt-sensitive hypertension without hyperkalemia that is characterized by upregulation of NCC, elevation of ß/?ENaC, but not aENaC, and a normal Na+/K+ balance maintained by downregulation of ENaC activity and upregulation of ROMK. [ABSTRACT FROM AUTHOR]

Objectives: Transient heat stress (HS) application during experimental ex vivo lung perfusion (EVLP) of warm ischaemic (WI) rat lungs produces a range of therapeutic benefits. Here, we explored whether different EVLP durations after HS application would influence its therapeutic effects., Methods: In protocol 1, WI rat lungs were exposed to HS (41.5°C, 60-90 min EVLP), and EVLP was maintained for 3, 4.5 or 6 h (n = 5/group), followed by physiological measurements (compliance, oedema, oxygenation capacity). In protocol 2, WI rat lungs treated with (HS groups) or without HS (control groups) were maintained for 3 or 4.5 h EVLP (n = 5/group), followed by physiological evaluation and measurements (lung tissue) of heat shock proteins (HSP70, HSP27, HS90, GRP78), endogenous proteins (surfactant protein-D, CC16, platelet endothelial cell adhesion molecule-1), anti-apoptotic (Bcl2, Bcl-xL) and pro-apoptotic proteins (Bcl2-associated X protein, CCAAT/enhancer binding-protein homologous protein), antioxidant enzymes (heme-oxygenase-1, nicotinamide di-phospho-nucleotide dehydrogenase quinone-1) and nitrotyrosine (oxidative stress biomarker)., Results: In protocol 1, physiological variables were stable after 3 and 4.5 h but deteriorated after 6 h. In protocol 2, at 3 h EVLP, HS-treated lungs differed from controls by higher expression of HSP70 and heme-oxygenase-1, and lower CC16 expression. In contrast, at 4.5 h EVLP, HS-treated lungs displayed improved physiology, higher levels of all HSPs, preserved or increased expression of surfactant protein-D, CC-16 and platelet endothelial cell adhesion molecule-1, increased antioxidant and anti-apoptotic proteins, and reduced pro-apoptotic proteins and nitrotyrosine., Conclusions: The protective effects of HS application during EVLP of WI-damaged rat lungs strictly depend on the duration of post-HS recovery. An EVLP duration of 4.5 h appears to optimize the therapeutic potential of HS, while maintaining lungs in a stable physiological state., (© The Author(s) 2025. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery.)

Introduction: The pulmonary endothelium is the primary target of lung ischemia-reperfusion injury leading to primary graft dysfunction after lung transplantation. We hypothesized that treating damaged rat lungs by a transient heat stress during ex-vivo lung perfusion (EVLP) to elicit a pulmonary heat shock response could protect the endothelium from severe reperfusion injury., Methods: Rat lungs damaged by 1h warm ischemia were reperfused on an EVLP platform for up to 6h at a constant temperature (T°) of 37°C (EVLP 37°C group), or following a transient heat stress (HS) at 41.5°C from 1 to 1.5h of EVLP (EVLP HS group). A group of lungs exposed to 1h EVLP only (pre-heating conditions) was added as control (Baseline group). In a first protocol, we measured lung heat sock protein expression (HSP70, HSP27 and Hsc70) at selected time-points (n=5/group at each time). In a second protocol, we determined (n=5/group) lung weight gain (edema), pulmonary compliance, oxygenation capacity, pulmonary artery pressure (PAP) and vascular resistance (PVR), the expression of PECAM-1 (CD31) and phosphorylation status of Src-kinase and VE-cadherin in lung tissue, as well as the release in perfusate of cytokines (TNFα, IL-1β) and endothelial biomarkers (sPECAM, von Willebrand Factor -vWF-, sE-selectin and sICAM-1). Histological and immunofluorescent studies assessed perivascular edema and formation of 3-nitrotyrosine (a marker of peroxinitrite) in CD31 lung endothelium., Results: HS induced an early (3h) and persisting expression of HSP70 and HSP27, without influencing Hsc70. Lungs from the EVLP 37°C group developed massive edema, low compliance and oxygenation, elevated PAP and PVR, substantial release of TNFα, IL-1β, s-PECAM, vWF, E-selectin and s-ICAM, as well as significant Src-kinase activation, VE-cadherin phosphorylation, endothelial 3-NT formation and reduced CD31 expression. In marked contrast, all these alterations were either abrogated or significantly attenuated by HS treatment., Conclusion: The therapeutic application of a transient heat stress during EVLP of damaged rat lungs reduces endothelial permeability, attenuates pulmonary vasoconstriction, prevents src-kinase activation and VE-cadherin phosphorylation, while reducing endothelial peroxinitrite generation and the release of cytokines and endothelial biomarkers. Collectively, these data demonstrate that therapeutic heat stress may represent a promising strategy to protect the lung endothelium from severe reperfusion injury., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Parapanov, Debonneville, Allouche, Lugrin, Rodriguez-Caro, Liaudet and Krueger.)

Blood pressure (BP) follows a circadian pattern that rises during the active phase of the day (morning surge) and decreases during the inactive (night dipping) phase of the day. The morning surge coincides with increased circulating glucocorticoids and aldosterone, ligands for glucocorticoid receptors and mineralocorticoid receptors, respectively. Serum- and glucocorticoid-induced kinase 1 ( SGK1 ), a clock-controlled and glucocorticoid receptor- and mineralocorticoid receptor-induced gene, plays a role in BP regulation in human and animal models. However, the role of SGK1 in BP circadian regulation has not yet been demonstrated. Using telemetry, we analyzed BP in the inducible renal tubule-specific Sgk1 Pax8/LC1 model under basal K + diet (1% K + ) and high-K + diet (HKD; 5% K + ). Our data revealed that, under basal conditions, renal SGK1 plays a minor role in BP regulation; however, after 1 wk of HKD, Sgk1 Pax8/LC1 mice exhibited significant defects in diastolic BP (DBP), including a blunted surge, a decreased amplitude, and reduced day/night differences. After prolonged HKD (7 wk), Sgk1 Pax8/LC1 ) the kidney plays a significant role in setting the BP circadian rhythm; 1 ) the kidney plays a significant role in setting the BP circadian rhythm; 2 ) renal tubule SGK1 mediates the BP surge and, thus, the day/night BP difference; 3 ) long-term renal SGK1 deletion results in lower BP in mutant compared with control mice; and 4 ) renal SGK1 indirectly regulates pulse pressure due to compensatory alterations in aldosterone levels. NEW & NOTEWORTHY Dysregulation of blood pressure (BP) circadian rhythm is associated with metabolic, cardiovascular, and kidney diseases. Our study provides experimental evidence demonstrating, for the first time, that renal tubule serum- and glucocorticoid-induced kinase 1 ( SGK1 ) plays an essential role in inducing the BP surge. Inhibitors and activators of SGK1 signaling are parts of several therapeutic strategies. Our findings highlight the importance of the drug intake timing to be in phase with SGK1 function to avoid dysregulation of BP circadian rhythm.

Ex vivo lung perfusion (EVLP) may serve as a platform for the pharmacologic repair of lung grafts before transplantation (LTx). We hypothesized that EVLP could also permit nonpharmacologic repair through the induction of a heat shock response, which confers stress adaptation via the expression of heat shock proteins (HSPs). Therefore, we evaluated whether transient heat application during EVLP (thermal preconditioning [TP]) might recondition damaged lungs before LTx. TP was performed during EVLP (3 hours) of rat lungs damaged by warm ischemia by transiently heating (30 minutes, 41.5 °C) the EVLP perfusate, followed by LTx (2 hours) reperfusion. We also assessed the TP (30 minutes, 42 °C) during EVLP (4 hours) of swine lungs damaged by prolonged cold ischemia. In rat lungs, TP induced HSP expression, reduced nuclear factor κB and inflammasome activity, oxidative stress, epithelial injury, inflammatory cytokines, necroptotic death signaling, and the expression of genes involved in innate immune and cell death pathways. After LTx, heated lungs displayed reduced inflammation, edema, histologic damage, improved compliance, and unchanged oxygenation. In pig lungs, TP induced HSP expression, reduced oxidative stress, inflammation, epithelial damage, vascular resistance, and ameliorated compliance. Collectively, these data indicate that transient heat application during EVLP promotes significant reconditioning of damaged lungs and improves their outcomes after transplantation., Competing Interests: Disclosure The authors of this manuscript have no conflicts of interest to disclose as described by the American Journal of Transplantation., (Copyright © 2023. Published by Elsevier Inc.)

Background: Hemoadsorption (HA) might mitigate the systemic inflammatory response associated with post-cardiac arrest syndrome (PCAS) and improve outcomes. Here, we investigated the feasibility, safety and efficacy of HA with CytoSorb ® in cardiac arrest (CA) survivors at risk of PCAS., Methods: In this pilot randomized controlled trial, we included patients admitted to our intensive care unit following CA and likely to develop PCAS: required norepinephrine (> 0.2 µg/kg/min), and/or had serum lactate > 6 mmol/l and/or a time-to-return of spontaneous circulation (ROSC) > 25 min. Those requiring ECMO or renal replacement therapy were excluded. Eligible patients were randomly allocated to either receive standard of care (SOC) or SOC plus HA. Hemoadsorption was performed as stand-alone therapy for 24 h, using CytoSorb ® and regional heparin-protamine anticoagulation. We collected feasibility, safety and clinical data as well as serial plasma cytokines levels within 72 h of randomization., Results: We enrolled 21 patients, of whom 16 (76%) had out-of-hospital CA. Median (IQR) time-to-ROSC was 30 (20, 45) minutes. Ten were assigned to the HA group and 11 to the SOC group. Hemoadsorption was initiated in all patients allocated to the HA group within 18 (11, 23) h of ICU admission and conducted for a median duration of 21 (14, 24) h. The intervention was well tolerated except for a trend for a higher rate of aPTT elevation (5 (50%) vs 2 (18%) p = 0.18) and mild (100-150 G/L) thrombocytopenia at day 1 (5 (50%) vs 2 (18%) p = 0.18). Interleukin (IL)-6 plasma levels at randomization were low (< 100 pg/mL) in 10 (48%) patients and elevated (> 1000 pg/mL) in 6 (29%). The median relative reduction in IL-6 at 48 h was 75% (60, 94) in the HA group versus 5% (- 47, 70) in the SOC group (p = 0.06)., Conclusions: In CA survivors at risk of PCAS, HA was feasible, safe and was associated with a nonsignificant reduction in cytokine plasma levels. Future trials are needed to further define the role of HA after CA. Those studies should include cytokine assessment to enrich the study population., Trial Registration: NCT03523039, registered 14 May 2018., (© 2023. The Author(s).)

Background: Ex vivo lung perfusion (EVLP) may allow therapeutic reconditioning of damaged lung grafts before transplantation. This study aimed to develop relevant rat models of lung damage to study EVLP therapeutic reconditioning for possible translational applications., Methods: Lungs from 31 rats were exposed to cold ischemia (CI) or warm ischemia (WI), inflated at various oxygen fractions (FiO 2 ), followed by 3 h EVLP. Five groups were studied as follow: (1) C21 (control): 3 h CI (FiO 2 0.21); (2) C50: 3 h CI (FiO 2 0.5); (3) W21: 1 h WI, followed by 2 h CI (FiO 2 0.21); (4) W50: 1 h WI, followed by 2 h CI (FiO 2 0.5); and (5) W2h: 2 h WI, followed by 1 h CI (FiO 2 0.21). Following 3 h EVLP, we measured static pulmonary compliance (SPC), pulmonary vascular resistance, lung weight gain (edema), oxygenation capacity (differential partial pressure of oxygen), and protein carbonyls in lung tissue (oxidative stress), as well as lactate dehydrogenase (LDH, lung injury), nitrotyrosine (nitro-oxidative stress), interleukin-6 (IL-6, inflammation), and proteins (permeability edema) in bronchoalveolar lavage (BAL). Perivascular edema was quantified by histology., Results: No significant alterations were noted in C21 and C50 groups. W21 and W50 groups had reduced SPC and disclosed increased weight gain, BAL proteins, nitrotyrosine, and LDH. These changes were more severe in the W50 group, which also displayed greater oxidative stress. In contrast, both W21 and W50 showed comparable perivascular edema and BAL IL-6. In comparison with the other WI groups, W2h showed major weight gain, perivascular edema, SPC reduction, drop of differential partial pressure of oxygen, and massive increases of BAL LDH and proteins but comparable increase of IL-6 and biomarkers of oxidative stress., Conclusions: These models of lung damage of increasing severity might be helpful to evaluate new strategies for EVLP therapeutic reconditioning. A model combining 1 h WI and inflation at FiO 2 of 0.5 seems best suited for this purpose by reproducing major alterations of clinical lung ischemia-reperfusion injury., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2022 The Author(s). Transplantation Direct. Published by Wolters Kluwer Health, Inc.)

SIRT7 is a NAD + -dependent deacetylase that controls important aspects of metabolism, cancer, and bone formation. However, the molecular targets and functions of SIRT7 in the kidney are currently unknown. In silico analysis of kidney transcripts of the BXD murine genetic reference population revealed a positive correlation between Sirt7 and Slc12a7 mRNA expression, suggesting a link between the corresponding proteins that these transcripts encode, SIRT7, and the K-Cl cotransporter KCC4, respectively. Here, we find that protein levels and activity of heterologously expressed KCC4 are significantly modulated depending on its acetylation status in Xenopus laevis oocytes. Moreover, SIRT7 interacts with KCC4 in a NAD + -dependent manner and increases its stability and activity in HEK293 cells. Interestingly, metabolic acidosis increases SIRT7 expression in kidney, as occurs with KCC4. In contrast, total SIRT7-deficient mice present lower KCC4 expression and an exacerbated metabolic acidosis than wild-type mice during an ammonium chloride challenge. Altogether, our data suggest that SIRT7 interacts with, stabilizes and modulates KCC4 activity through deacetylation, and reveals a novel role for SIRT7 in renal physiology., (© 2021 The Authors.)

Ex vivo lung perfusion (EVLP) with pharmacological reconditioning may increase donor lung utilization for transplantation (LTx). 3-Aminobenzamide (3-AB), an inhibitor of poly(ADP-ribose) polymerase (PARP), reduces ex vivo lung injury in rat lungs damaged by warm ischemia (WI). Here we determined the effects of 3-AB reconditioning on graft outcome after LTx. Three groups of donor lungs were studied: Control (Ctrl): 1 hour WI + 3 hours cold ischemia (CI) + LTx; EVLP: 1 hour WI + 3 hours EVLP + LTx; EVLP + 3-AB: 1 hour WI + 3 hours EVLP + 3-AB (1 mg . mL -1 ) + LTx. Two hours after LTx, we determined lung graft compliance, edema, histology, neutrophil counts in bronchoalveolar lavage (BAL), mRNA levels of adhesion molecules within the graft, as well as concentrations of interleukin-6 and 10 (IL-6, IL-10) in BAL and plasma. 3-AB reconditioning during EVLP improved compliance and reduced lung edema, neutrophil infiltration, and the expression of adhesion molecules within the transplanted lungs. 3-AB also attenuated the IL-6/IL-10 ratio in BAL and plasma, supporting an improved balance between pro- and anti-inflammatory mediators. Thus, 3-AB reconditioning during EVLP of rat lung grafts damaged by WI markedly reduces inflammation, edema, and physiological deterioration after LTx, supporting the use of PARP inhibitors for the rehabilitation of damaged lungs during EVLP., (© 2019 The American Society of Transplantation and the American Society of Transplant Surgeons.)

Chronic glucocorticoid infusion impairs NCC activity and induces a non-dipping profile in mice, suggesting that glucocorticoids are essential for daily blood pressure variations. In this paper, we studied mice lacking the renal tubular glucocorticoid receptor (GR) in adulthood (GR knockouts, Nr3c1 Pax8/LC1 ). Upon standard salt diet, Nr3c1 mice exhibit a transient but significant reduction in the activity of NCC and expression of NHE3 and NKCC2 accompanied by significant increased Spak activity. This is followed by transiently increased urinary sodium excretion and higher plasma aldosterone concentrations. Plasma corticosterone levels and 11βHSD2 mRNA expression and activity in the whole kidney remain unchanged. High salt diet does not affect whole body Na Pax8/LC1 mice grow normally, but show reduced NCC activity despite normal plasma aldosterone levels. Following diet switch to low sodium, Nr3c1 balance and NCC activity is not reduced, but leads to a significant increase in diastolic blood pressure dipping in Pax8/LC1 mice exhibit a transient but significant reduction in the activity of NCC and expression of NHE3 and NKCC2 accompanied by significant increased Spak activity. This is followed by transiently increased urinary sodium excretion and higher plasma aldosterone concentrations. Plasma corticosterone levels and 11βHSD2 mRNA expression and activity in the whole kidney remain unchanged. High salt diet does not affect whole body Na + and/or K + balance and NCC activity is not reduced, but leads to a significant increase in diastolic blood pressure dipping in Nr3c1 Pax8/LC1 mice. When high sodium treatment is followed by 48 h of darkness, NCC abundance is reduced in knockout mice although activity is not different. Our data show that upon Na + restriction renal tubular GR-deficiency transiently affects Na + handling and transport pathways. Overall, upon standard, low Na + and high Na + diet exposure Na + and K + balance is maintained as evidenced by normal plasma and urinary Na + and K + and aldosterone concentrations.

Objective: The aim of the study was to search for mutations of SCNN1B and SCNN1G in an Italian family with apparently dominant autosomal transmission of a clinical phenotype consistent with Liddle's syndrome., Methods: Genetic analysis was performed in the proband, his relatives, and 100 control subjects. To determine the functional role of the mutation identified in the proband, we expressed the mutant or wild-type epithelial sodium channel in Xenopus laevis oocytes., Results: A novel point mutation, causing an expected substitution of a leucine residue for the second proline residue of the conserved PY motif (PPP x Y) of the beta subunit was identified in the proband. The functional expression of the mutant epithelial sodium channel in X. laevis oocytes showed a three-fold increase in the amiloride-sensitive current as compared with that of the wild-type channel., Conclusion: This newly identified mutation adds to other missense mutations of the PY motif of the beta subunit of the epithelial sodium channel, thus confirming its crucial role in the regulation of the epithelial sodium channel. To our knowledge, this is the first report of Liddle's syndrome in the Italian population, confirmed by genetic and functional analysis, with the identification of a gain-of-function mutation not previously reported.