Search

Abstract Hepatoblastoma, the most prevalent pediatric liver cancer, almost always carries a WNT-activating CTNNB1 mutation, yet exhibits notable molecular heterogeneity. To characterize this heterogeneity and identify novel targeted therapies, we perform comprehensive analysis of hepatoblastomas and tumor-derived organoids using single-cell RNA-seq/ATAC-seq, spatial transcriptomics, and high-throughput drug profiling. We identify two distinct tumor epithelial signatures: hepatic ‘fetal’ and WNT-high ‘embryonal’, displaying divergent WNT signaling patterns. The fetal group is enriched for liver-specific WNT targets, while the embryonal group is enriched in canonical WNT target genes. Gene regulatory network analysis reveals enrichment of regulons related to hepatic functions such as bile acid, lipid and xenobiotic metabolism in the fetal subtype but not in the embryonal subtype. In addition, the dichotomous expression pattern of the transcription factors HNF4A and LEF1 allows for a clear distinction between the fetal and embryonal tumor cells. We also perform high-throughput drug screening using patient-derived tumor organoids and identify sensitivity to HDAC inhibitors. Intriguingly, embryonal and fetal tumor organoids are sensitive to FGFR and EGFR inhibitors, respectively, indicating a dependency on EGF/FGF signaling in hepatoblastoma tumorigenesis. In summary, our data uncover the molecular and drug sensitivity landscapes of hepatoblastoma and pave the way for the development of targeted therapies.

Background:. More than 65% of patients with end-stage renal disease (ESRD) use arteriovenous fistulas (AVFs) for hemodialysis. The increasing incidence of comorbid ESRD and obesity (body mass index, >35 kg/m2) precludes patients from kidney transplantation, resulting in a need for long-term, durable AVF access. Compared with traditional superficialization techniques for overlying adiposity, liposuction is minimally invasive and well-tolerated, allowing for earlier fistula use with lower complications. We present the detailed surgical technique for superficialization of AVFs using liposuction. Methods:. Fourteen patients with well-matured but difficult-access fistulas due to adiposity were selected. Preoperative ultrasound mapped depth of fistulas. Tumescent liposuction was completed in a cross-hatched manner. Intraoperative ultrasound confirmed cannula positioning and measured fistula depth. A palpable thrill remained throughout superficialization. Cannulation began 4 weeks postoperatively. Results:. Mean access depth preoperatively was 10.8 mm (8–15 mm), immediately postoperative was 7 mm (6–9 mm), and at 4 weeks was 5.3 mm (4–8 mm). The average usable access length was 12.7 cm (10–15 cm) after surgery. Thirteen fistulas were successfully accessed after liposuction superficialization. All patients were discharged home the same day. There were no postoperative infections or hemorrhage. Conclusions:. Early experience with liposuction for superficialization of deep hemodialysis access is promising. This innovative solution has the possibility to improve outcomes and quality of life for patients living with ESRD and obesity. Our experience shows that this is a safe and effective technique to increase patient eligibility, enable successful and early cannulation, and decrease recovery time.

Purpose Elevated dopamine synthesis capacity is part of the pathophysiology of schizophrenia thought to underlie psychosis. Drugs that reduce this phenomenon could thus be potential treatments for these disorders. In this study, we evaluated the ability of the trace amine-associated receptor 1 (TAAR1) partial agonist ralmitaront to reduce presynaptic dopamine synthesis capacity. Procedures Ralmitaront (3 mg/kg, i.p.), a TAAR1 partial agonist, was evaluated using [ 18 F]DOPA PET for its ability to modulate presynaptic dopamine synthesis capacity in naïve mice as well as mice in an induced hyperdopaminergic state following acute cocaine administration (20 mg/kg, i.p.). Results Cocaine treatment on its own did not induce elevated dopamine synthesis capacity when compared to the control group. Pretreatment with ralmitaront significantly reduced dopamine synthesis capacity when given either alone (44%) or in combination with the psychostimulant cocaine (50%) when compared to the control group. Conclusions The TAAR1 agonist ralmitaront reduces striatal dopamine synthesis capacity, indexed as Ki Mod , both in naïve animals and when given prior to acute cocaine. This indicates the potential of TAAR1 agonism to address disorders characterized by striatal hyperdopaminergia.

Plasma-enhanced chemical vapor deposition (PECVD) was used to obtain several graphite nanowall (GNW)-type films at different deposition times on silicon and copper to achieve various thicknesses of carbonic films for the development of electrochemical sensors for the detection of anthracene. The PECVD growth time varied from 15 min to 30 min to 45 min, while scanning electron microscopy (SEM) confirmed the changes in the thickness of the GNW films, revealing a continuous increase in the series. X-ray diffraction (XRD) analysis revealed that the crystallinity of the GNW film samples increased with increasing crystallite size and decreasing dislocation density as the deposition time increased. Electrochemical characterization of the GNW-based electrodes indicated that the electroactive area and heterogeneous electron transfer rate constant were greater for the GNW 45 min film in the carbonic material series. We present the transfer of GNW films on flexible polyethylene substrates for achieving flexible electrochemical sensors for further use in anthracene determination. The flexible GNW-based electrodes were investigated using differential pulse voltammetry (DPV) in the presence of anthracene. The results showed that the highest sensitivity in anthracene detection was provided by the sensor with the GNW film obtained after 45 min of PECVD growth. The optimization of the GNW film thickness for the development of flexible electrochemical sensors on polyethylene substrates represents a successful approach for enhancing the electrochemical performance of carbonic materials.

This study presents a novel methodology for the rapid on-site detection of tannic acid (TA), a prevalent organic contaminant in various natural environments, notably in plant-derived sources. The proposed approach involves the development of a compact integrated electrochemical sensor incorporating a nanozyme system. Specifically, this system comprises Fe2O3 nanoparticles (NPs) embedded within a chitosan (CS) matrix, immobilized onto a sulfur-doped graphene (S-Gr) substrate deposited on a gold electrode (AuE). The Fe2O3NPs exhibit peroxidase-like artificial enzyme activity, contributing to exceptional stability and catalytic efficiency in TA oxidation processes. Additionally, the CS matrix acts as a stabilizing agent, enhancing the performance and recyclability of the nanozyme. Furthermore, the S-Gr nanomaterial facilitates rapid electron transfer, leading to heightened sensitivity and prompt response times. The integration of these advanced nanomaterials with a microfabricated electrode presents an economically feasible, reliable, and effective solution for TA detection, with promising prospects for large-scale deployment and environmental monitoring. The Fe2O3CS-S-Gr/AuE sensing system demonstrates a calculated limit of detection (LOD) of 3.6 × 10−3 µM and an increased sensitivity of 0.2 µA×µM−1, with a wide linear concentration range spanning from 0.01 to 1000 µM for TA detection. Notably, the recovery values obtained for surface water samples fall within the range of 97.7 % to 99.5 %, indicating strong agreement with results derived from the standard method, UHPLC-MS/MS.

Background: In recent years, cardiac dysfunction in childhood cancer survivors has become an important issue. Studies are focusing on identifying means for the early identification of patients at risk. Considering this, our study aims to investigate 24-hour Holter electrocardiogram (ECG) repolarization changes throughout doxorubicin (DOX) and cyclophosphamide (CPM) administration in pediatric patients treated for acute lymphoblastic leukemia (ALL). Methods: This was an investigator-driven, single-center, prospective, observational study. Enrolled children had a baseline bedside ECG examination performed before starting chemotherapy (T0). Serial Holter ECG examinations were conducted at three moments during their treatment protocol: day 8 (T1), day 29 (T2), and day 36 (T3). This study evaluated several ECG repolarization parameters, such as the QT interval, corrected QT interval (QTc), and QTc dispersion, as well as ST segment variations. Results: We evaluated 37 children diagnosed with ALL. The T0 examination revealed that over a third of patients had a resting heart rate (HR) outside the normal range for their age and sex. During chemotherapy, statistically significant increases in both HR as well as QT and QTc dispersion values were noticed, especially during the first DOX administration. What is more, a significant increase in the percentage of patients with ST segment depression from T1 to T2 and T3 was noticed. Rhythm disturbances were rare in the study population, with only a few patients presenting ventricular or supraventricular extrasystoles. Conclusions: This study reveals silent repolarization changes occurring early during anticancer treatment in children treated for ALL. These findings could aid in a better understanding of the cardiac toxicity mechanism, and they could potentially improve cardiac risk stratification for oncologic patients. Because of the small number of patients, our results need to be validated by larger studies.

In vivo, psilocybin is rapidly dephosphorylated to psilocin which induces psychedelic effects by interacting with the 5-HT2A receptor. Psilocin primarily undergoes glucuronidation or conversion to 4-hydroxyindole-3-acetic acid (4-HIAA). Herein, we investigated psilocybin’s metabolic pathways in vitro and in vivo, conducting a thorough analysis of the enzymes involved. Metabolism studies were performed using human liver microsomes (HLM), cytochrome P450 (CYP) enzymes, monoamine oxidase (MAO), and UDP-glucuronosyltransferase (UGT). In vivo, metabolism was examined using male C57BL/6J mice and human plasma samples. Approximately 29% of psilocin was metabolized by HLM, while recombinant CYP2D6 and CYP3A4 enzymes metabolized nearly 100% and 40% of psilocin, respectively. Notably, 4-HIAA and 4-hydroxytryptophol (4-HTP) were detected with HLM but not with recombinant CYPs. MAO-A transformed psilocin into minimal amounts of 4-HIAA and 4-HTP. 4-HTP was only present in vitro. Neither 4-HIAA nor 4-HTP showed relevant interactions at assessed 5-HT receptors. In contrast to in vivo data, UGT1A10 did not extensively metabolize psilocin in vitro. Furthermore, two putative metabolites were observed. N-methyl-4-hydroxytryptamine (norpsilocin) was identified in vitro (CYP2D6) and in mice, while an oxidized metabolite was detected in vitro (CYP2D6) and in humans. However, the CYP2D6 genotype did not influence psilocin plasma concentrations in the investigated study population. In conclusion, MAO-A, CYP2D6, and CYP3A4 are involved in psilocin’s metabolism. The discovery of putative norpsilocin in mice and oxidized psilocin in humans further unravels psilocin’s metabolism. Despite limitations in replicating phase II metabolism in vitro, these findings hold significance for studying drug-drug interactions and advancing research on psilocybin as a therapeutic agent.

The realization of a carbon-neutral civilization, which has been set as a goal for the coming decades, goes directly hand-in-hand with the need for an energy system based on renewable energies (REs). Due to the strong weather-related, daily, and seasonal fluctuations in supply of REs, suitable energy storage devices must be included for such energy systems. For this purpose, an energy system model featuring hybrid energy storage consisting of a hydrogen unit (for long-term storage) and a lithium-ion storage device (for short-term storage) was developed. With a proper design, such a system can ensure a year-round energy supply by using electricity generated by photovoltaics (PVs). In the energy system that was investigated, hydrogen (H2) was produced by using an electrolyser (ELY) with a PV surplus during the summer months and then stored in an H2 tank. During the winter, due to the lack of PV power, the H2 is converted back into electricity and heat by a fuel cell (FC). While the components of such a system are expensive, a resource- and cost-efficient layout is important. For this purpose, a Matlab/Simulink model that enabled an energy balance analysis and a component lifetime forecast was developed. With this model, the results of extensive parameter studies allowed an optimized system layout to be created for specific applications. The parameter studies covered different focal points. Several ELY and FC layouts, different load characteristics, different system scales, different weather conditions, and different load levels—especially in winter with variations in heating demand—were investigated.

This paper presents a model of an energy system for a private household extended by a lifetime prognosis. The energy system was designed for fully covering the year-round energy demand of a private household on the basis of electricity generated by a photovoltaic (PV) system, using a hybrid energy storage system consisting of a hydrogen unit and a lithium-ion battery. Hydrogen is produced with a Proton Exchange Membrane (PEM) electrolyser by PV surplus during the summer months and then stored in a hydrogen tank. Mainly during winter, in terms of lack of PV energy, the hydrogen is converted back into electricity and heat by a fuel cell. The model was created in Matlab/Simulink and is based on real input data. Heat demand was also taken into account and is covered by a heat pump. The simulation period is a full year to account for the seasonality of energy production and demand. Due to high initial costs, the longevity of such an energy system is of vital interest. Therefore, this model was extended by a lifetime prediction in order to optimize the dimensioning with the aim of lifetime extension of a hydrogen-based energy system. Lifetime influencing factors were identified on the basis of a literature review and were integrated in the model. An extensive parameter study was performed to evaluate different dimensionings regarding the energy balance and the lifetime of the three components, electrolyser, fuel cell and lithium-ion battery. The results demonstrate the benefits of a holistic modelling approach and enable a design optimization regarding the use of resources, lifetime and self-sufficiency of the system.

Background. Normothermic machine perfusion (NMP) is a promising modality for marginal donor kidneys. However, little is known about the effects of NMP on causing endothelial glycocalyx (eGC) injury. This study aims to evaluate the effects of NMP on eGC injury in marginal donor kidneys and whether this is affected by perfusion pressures and hematocrits. Methods. Porcine slaughterhouse kidneys (n = 6/group) underwent 35 min of warm ischemia. Thereafter, the kidneys were preserved with oxygenated hypothermic machine perfusion for 3 h. Subsequently, 4 h of NMP was applied using pressure-controlled perfusion with an autologous blood-based solution containing either 12%, 24%, or 36% hematocrit. Pressures of 55, 75, and 95 mm Hg were applied in the 24% group. Perfusate, urine, and biopsy samples were collected to determine both injury and functional parameters. Results. During NMP, hyaluronan levels in the perfusate increased significantly (P < 0.0001). In addition, the positivity of glyco-stained glycocalyx decreased significantly over time, both in the glomeruli (P = 0.024) and peritubular capillaries (P = 0.003). The number of endothelial cells did not change during NMP (P = 0.157), whereas glomerular endothelial expression of vascular endothelial growth factor receptor-2 decreased significantly (P < 0.001). Microthrombi formation was significantly increased after NMP. The use of different pressures and hematocrits did not affect functional parameters during perfusion. Conclusions. NMP is accompanied with eGC and vascular endothelial growth factor receptor-2 loss, without significant loss of endothelial cells. eGC loss was not affected by the different pressures and hematocrits used. It remains unclear whether endothelial injury during NMP has harmful consequences for the transplanted kidney.

Abstract Background Immunotherapy with immune checkpoint inhibitors (ICI) has revolutionized cancer therapy. However, therapeutic targeting of inhibitory T cell receptors such as PD-1 not only initiates a broad immune response against tumors, but also causes severe adverse effects. An ideal future stratified immunotherapy would interfere with cancer-specific cell surface receptors only. Methods To identify such candidates, we profiled the surface receptors of the NCI-60 tumor cell panel via flow cytometry. The resulting surface receptor expression data were integrated into proteomic and transcriptomic NCI-60 datasets applying a sophisticated multiomics multiple co-inertia analysis (MCIA). This allowed us to identify surface profiles for skin, brain, colon, kidney, and bone marrow derived cell lines and cancer entity-specific cell surface receptor biomarkers for colon and renal cancer. Results For colon cancer, identified biomarkers are CD15, CD104, CD324, CD326, CD49f, and for renal cancer, CD24, CD26, CD106 (VCAM1), EGFR, SSEA-3 (B3GALT5), SSEA-4 (TMCC1), TIM1 (HAVCR1), and TRA-1-60R (PODXL). Further data mining revealed that CD106 (VCAM1) in particular is a promising novel immunotherapeutic target for the treatment of renal cancer. Conclusion Altogether, our innovative multiomics analysis of the NCI-60 panel represents a highly valuable resource for uncovering surface receptors that could be further exploited for diagnostic and therapeutic purposes in the context of cancer immunotherapy.

Background and aimsPrecision-cut tissue slices (PCTS) are widely used as an ex vivo culture tissue culture technique to study pathogeneses of diseases and drug activities in organs in vitro. A novel application of the PCTS model may be in the field of translational research into cholangiopathies such as biliary atresia. Therefore, the aim of this study was to apply the precision-cut slice technique to human bile duct and gallbladder tissue.MethodsCystic duct and gallbladder tissue derived from patients undergoing a cholecystectomy were collected, preserved and used for preparation of precision-cut cystic duct slices (PCCDS) and precision-cut gallbladder slices (PCGS). The PCCDS and PCGS were prepared using a mechanical tissue slicer and subsequently incubated for 24 and 48 h respectively in William's Medium E (WME) culture medium. Viability was assessed based on ATP/protein content and tissue morphology [hematoxylin and eosin (H&E) staining].ResultsIt was shown that viability, assessed by the ATP/protein content and morphology, of the PCCDS (n = 8) and PCGS (n = 8) could be maintained over the 24 and 48 h incubation period respectively. ATP/protein content of the PCCDS increased significantly from 0.58 ± 0.13 pmol/µg at 0 h to 2.4 ± 0.29 pmol/µg after 24 h incubation (P = .0003). A similar significant increase from 0.94 ± 0.22 pmol/µg at 0 h to 3.7 ± 0.41 pmol/µg after 24 h (P = .0005) and 4.2 ± 0.47 pmol/µg after 48 h (P = .0002) was observed in the PCGS. Morphological assessment of the PCCDS and PCGS showed viable tissue at 0 h and after 24 and 48 h incubation respectively.ConclusionThis study is the first to report on the use of the PCTS model for human gallbladder and cystic duct tissue. PCCDS and PCGS remain viable for an incubation period of at least 24 h, which makes them suitable for research purposes in the field of cholangiopathies, including biliary atresia.

Summary: Background: European data pre-2019 suggest statin monotherapy is the most common approach to lipid management for preventing cardiovascular (CV) events, resulting in only one-fifth of high- and very high-risk patients achieving the 2019 ESC/EAS recommended low-density lipoprotein cholesterol (LDL-C) goals. Whether the treatment landscape has evolved, or gaps persist remains of interest. Methods: Baseline data are presented from SANTORINI, an observational, prospective study that documents the use of lipid-lowering therapies (LLTs) in patients ≥18 years at high or very high CV risk between 2020 and 2021 across primary and secondary care settings in 14 European countries. Findings: Of 9602 enrolled patients, 9044 with complete data were included (mean age: 65.3 ± 10.9 years; 72.6% male). Physicians reported using 2019 ESC/EAS guidelines as a basis for CV risk classification in 52.0% (4706/9044) of patients (overall: high risk 29.2%; very high risk 70.8%). However, centrally re-assessed CV risk based on 2019 ESC/EAS guidelines suggested 6.5% (308/4706) and 91.0% (4284/4706) were high- and very high-risk patients, respectively. Overall, 21.8% of patients had no documented LLTs, 54.2% were receiving monotherapy and 24.0% combination LLT. Median (interquartile range [IQR]) LDL-C was 2.1 (1.6, 3.0) mmol/L (82 [60, 117] mg/dL), with 20.1% of patients achieving risk-based LDL-C goals as per the 2019 ESC/EAS guidelines. Interpretation: At the time of study enrolment, 80% of high- and very high-risk patients failed to achieve 2019 ESC/EAS guidelines LDL-C goals. Contributory factors may include CV risk underestimation and underutilization of combination therapies. Further efforts are needed to achieve current guideline-recommended LDL-C goals. Trial registration: ClinicalTrials.gov Identifier: NCT04271280. Funding: This study is funded by Daiichi Sankyo Europe GmbH, Munich, Germany.

Hepatitis B vaccination is recommended in all patients with end-stage kidney disease (ESKD). However, only 50–60% of these patients achieve protective antibody levels if immunized after starting dialysis. Strategies to overcome this low seroconversion rate include a 6-month vaccination schedule starting earlier [chronic kidney disease (CKD) stage 4 and 5] to ensure immunity when patients progress to ESKD. We conducted a quality improvement program to immunize pre-dialysis patients. Patients who were found to have a negative baseline serology with a negative hepatitis B surface antibody level (HBsAb) were offered vaccination on a 6-month schedule (0, 1 and 6 months) with one of two available vaccines within the VA system (Recombivax™ or Engerix™). HBsAb titers were checked 3–4 months later, and titers ≥ 12 mIU/mL were indicative of immunity at VA. Patients who did not seroconvert were offered a repeat schedule of three more doses. We screened 198 patients (187 males and 11 females) with CKD 4 and 5 [glomerular filtration rate (GFR) < 29 mL/min/1.73 m2]. The median age of this cohort was 72 years (range 38–92 years). During the study period of 5 years (2015–2020), 10 patients were excluded since their GFR had improved to more than 30 mL/min/1.73 m2, 24 others had baseline immunity and 2 refused vaccination. The hepatitis B vaccination series was not started on 106 patients. Of the remaining 56, 12 patients progressed to ESKD and started dialysis before completion of the vaccination schedule, 6 expired and 1 did not come to clinic in 2020 due to the pandemic. Of the 37 patients who completed the vaccination schedule, 16 achieved seroconversion with adequate HBsAb titers, 10 did not develop immunity despite a second hepatitis B vaccination series, while 11 did not get a second series. Given the low seroconversion rate, albeit in a small cohort, vaccination should be considered in patients with earlier stages of CKD. Other options include studies on FDA approved vaccines of shorter duration. We plan to increase awareness among nephrologists, patients and nursing staff about the importance of achieving immunity against hepatitis B.

Background. Hemangioblastomas are considered rare tumours that are located, in most cases, in the posterior cranial fossa. In most of cases, spinal hemangioblastomas are identified on the thoracic and cervical levels and can be associated in one-third of cases with von Hippel Lindau Syndrome. Material and methods. In this paper, we are presenting our clinic`s experience with spinal hemangioblastomas and the follow-up of the patients in the last 10 years. Results. In our study, we included six men and three women with a mean age of 45 years, ranging between 36 to 61 years. The mean hospitalization days was 17 days. Among nine patients, there were identified 12 tumors. Six patients presented cervical spine hemangioblastomas, two of them had thoracic spine lesions and in one case, the tumour was identified at the lumbar level. Furthermore, two patients presented multiple lesions. Two patients had been associated with von Hippel Lindau Syndrome. In all cases, surgery was the therapeutic approach. Discussions. Despite the rapid advance of technology and the new alternatives for the treatment of these lesions, surgical resection of hemangioblastomas remains the gold standard treatment. Conclusions. Hemangiolbastomas are benign tumours that can be associated with von Hippel Lindau. A whole-body scan is required to confirm or exclude this syndrome. Even though the surgical treatment for asymptomatic patients remains debatable, surgical intervention is the only treatment that can lead to the total removal of the tumour.

Background. Even in the 21st century, infections of the central nervous system and their secondary damage still represents a high mortality condition. Furthermore, brain abscesses (BA) are huge public health issues due to their high mortality and morbidity with high financial implications for the health care system. Material and methods. In this article, we want to present our clinic`s experience in the management of brain abscesses between 2012 and 2021. A 10 years retrospective study was performed in the 4th Neurosurgery Department of “Bagdasar-Arseni” Clinical Emergency Hospital in Bucharest Results. In the last 10 years, our clinic admitted 46 patients, 10 women and 36 men. The mean age was 47 years old. The most common symptom was headache (42 cases). In 39 cases, patients had solitary abscesses while seven patients presented multiple abscesses. The therapeutic approach was classic surgery in 37 cases. In four cases abscess drainage was performed and in five cases conservative treatment was used. At discharge, 31 patients were healed and seven patients died. Discussions. In BA management, antimicrobial therapy is crucial. Thus, identifying the pathogen is crucial for accurate antimicrobial treatment. Unfortunately, in too many cases, the agent remains unknown and empiric treatment is required. Aspiration of the abscesses proved to be safe and with similar results compared to classic surgery. Conclusions. Brain abscesses remain a high challenge to manage even in the 21st century. Broad-spectrum antibiotic treatment should be applied as soon as possible until the pathogen is identified and specific treatment will be used.

Abstract In kidney transplantation, microthrombi and fibrin deposition may lead to local perfusion disorders and subsequently poor initial graft function. Microthrombi are often regarded as donor-derived. However, the incidence, time of development, and potential difference between living donor kidneys (LDK) and deceased donor kidneys(DDK), remains unclear. Two open-needle biopsies, taken at preimplantation and after reperfusion, were obtained from 17 LDK and 28 DDK transplanted between 2005 and 2008. Paraffin-embedded sections were immunohistochemically stained with anti-fibrinogen antibody. Fibrin deposition intensity in peritubular capillaries(PTC) and glomeruli was categorized as negative, weak, moderate or strong and the number of microthrombi/mm2 was quantified. Reperfusion biopsies showed more fibrin deposition (20% to 100% moderate/strong, p

Abstract Background The mechanisms driving acute kidney injury (AKI) in critically ill COVID-19 patients are unclear. We collected kidney biopsies from COVID-19 AKI patients within 30 min after death in order to examine the histopathology and perform mRNA expression analysis of genes associated with renal injury. Methods This study involved histopathology and mRNA analyses of postmortem kidney biopsies collected from patients with COVID-19 (n = 6) and bacterial sepsis (n = 27). Normal control renal tissue was obtained from patients undergoing total nephrectomy (n = 12). The mean length of ICU admission-to-biopsy was 30 days for COVID-19 and 3–4 days for bacterial sepsis patients. Results We did not detect SARS-CoV-2 RNA in kidney biopsies from COVID-19-AKI patients yet lung tissue from the same patients was PCR positive. Extensive acute tubular necrosis (ATN) and peritubular thrombi were distinct histopathology features of COVID-19-AKI compared to bacterial sepsis-AKI. ACE2 mRNA levels in both COVID-19 (fold change 0.42, p = 0.0002) and bacterial sepsis patients (fold change 0.24, p

Over the coming years, AI could increasingly replace humans for making complex decisions because of the promise it holds for standardizing and debiasing decision-making procedures. Despite intense debates regarding algorithmic fairness, little research has examined how laypeople react when resource-allocation decisions are turned over to AI. We address this question by examining the role of perceived impartiality as a factor that can influence the acceptance of AI as a replacement for human decision-makers. We posit that laypeople attribute greater impartiality to AI than human decision-makers. Our investigation shows that people value impartiality in decision procedures that concern the allocation of scarce resources and that people perceive AI as more capable of impartiality than humans. Yet, paradoxically, laypeople prefer human decision-makers in allocation decisions. This preference reverses when potential human biases are made salient. The findings highlight the importance of impartiality in AI and thus hold implications for the design of policy measures.

A patient with cystic fibrosis (CF) with pancreatic insufficiency presented with jaundice due to an ampullary tumour. CF is known for a higher incidence of gastrointestinal malignancies. The patient suffered from pancreatic insufficiency. At computed tomography (CT), pancreatic lipomatosis with absence of the pancreatic duct was seen. This is uncommon, also in patients with CF. During surgery, a total pancreatectomy was performed, because there was no possibility to construct a duct to mucosa anastomosis due to the absence of the pancreatic duct and more importantly the pancreas was already afunctional. The presence of lipomatosis increases the risk of leakage at the pancreaticojejunal anastomosis. Therefore, it is important to take this phenomenon, in this case already visible on the preoperative CT scan, into account during the workup for surgery.

BackgroundThe role of the complement system in antibody-mediated rejection (ABMR) is insufficiently understood. We aimed to investigate the role of local and systemic complement activation in active (aABMR). We quantified complement activation markers, C3, C3d, and C5b-9 in plasma of aABMR, and acute T-cell mediated rejection (aTCMR), and non-rejection kidney transplant recipients. Intra-renal complement markers were analyzed as C4d, C3d, C5b-9, and CD59 deposition. We examined in vitro complement activation and CD59 expression on renal endothelial cells upon incubation with human leukocyte antigen antibodies.MethodsWe included 50 kidney transplant recipients, who we histopathologically classified as aABMR (n=17), aTCMR (n=18), and non-rejection patients (n=15).ResultsComplement activation in plasma did not differ across groups. C3d and C4d deposition were discriminative for aABMR diagnosis. Particularly, C3d deposition was stronger in glomerular (P

3,4,5-Trimethoxyphenethylamine (mescaline) is a psychedelic alkaloid found in peyote cactus. Related 4-alkoxy-3,5-dimethoxy-substituted phenethylamines (scalines) and amphetamines (3C-scalines) are reported to induce similarly potent psychedelic effects and are therefore potential novel therapeutics for psychedelic-assisted therapy. Herein, several pharmacologically uninvestigated scalines and 3C-scalines were examined at key monoamine targets in vitro. Binding affinity at human serotonergic 5-HT1A, 5-HT2A, and 5-HT2C, adrenergic α1A and α2A, and dopaminergic D2 receptors, rat and mouse trace amine-associated receptor 1 (TAAR1), and human monoamine transporters were assessed using target specific transfected cells. Furthermore, activation of human 5-HT2A and 5-HT2B receptors, and TAAR1 was examined. Generally, scalines and 3C-scalines bound with weak to moderately high affinity to the 5-HT2A receptor (Ki = 150–12,000 nM). 3C-scalines showed a marginal preference for the 5-HT2A vs the 5-HT2C and 5-HT1A receptors whereas no preference was observed for the scalines. Extending the 4-alkoxy substituent increased 5-HT2A and 5-HT2C receptors binding affinities, and enhanced activation potency and efficacy at the 5-HT2A but not at the 5-HT2B receptor. Introduction of fluorinated 4-alkoxy substituents generally increased 5-HT2A and 5-HT2C receptors binding affinities and increased the activation potency and efficacy at the 5-HT2A and 5-HT2B receptors. Overall, no potent affinity was observed at non-serotonergic targets. As observed for other psychedelics, scalines and 3C-scalines interacted with the 5-HT2A and 5-HT2C receptors and bound with higher affinities (up to 63-fold and 34-fold increase, respectively) when compared to mescaline.

ABSTRACT The plasma membrane (PM) must be overcome by viruses during entry and release. Furthermore, the PM represents the cellular communication compartment and the immune system interface. Hence, viruses have evolved sophisticated strategies to remodel the PM, for instance to avoid immune sensing and clearance of infected cells. We performed a comprehensive analysis of cell surface dysregulation by two human-pathogenic viruses, human cytomegalovirus (HCMV) and human immunodeficiency virus type 1 (HIV-1), in primary macrophages, which are classical antigen-presenting cells and orchestrators of the immune system. Scanning ion conductance microscopy revealed a loss of roughness and an overall smooth phenotype of HCMV-infected macrophages, in contrast to HIV-1 infection. This phenotype was also evident on the molecular level. When we screened for cell surface receptors modulated by HCMV, 42 of 332 receptors tested were up- or downregulated, whereas HIV-1 affected only 7 receptors. In particular CD164, CD84, and CD180 were targeted by HCMV. Mechanistically, HCMV induced transcriptional silencing of these receptors in an interferon (IFN)-independent manner, and expression was reduced not only by lab-adapted HCMV but also by clinical HCMV isolates. Altogether, our plasma membrane profiling of human macrophages provides clues to understand how viruses evade the immune system and identified novel cell surface receptors targeted by HCMV. IMPORTANCE The PM is a key component that viruses have to cope with. It is a barrier for infection and egress and is critically involved in antiviral immune signaling. We hence asked the question how two immunomodulatory viruses, HIV-1 and HCMV, dysregulate this compartment in infected macrophages, relevant in vivo targets of both viruses. We employed a contact-free microscopic technique to image the PM of infected cells and performed a phenotypic flow cytometry-based screen to identify receptor modulations on a molecular level. Our results show that HIV-1 and HCMV differentially manipulate the PM of macrophages. While HIV-1-mediated changes are relatively subtle, HCMV induces major alterations of the PM. We identify novel immune receptors manipulated by HCMV and define mechanisms of how HCMV interferes with receptor expression. Altogether, our study reveals differential strategies of how two human-pathogenic viruses manipulate infected cells and identifies potential novel pathways of HCMV immune evasion.

Angelman syndrome (AS) is a severe neurodevelopmental disorder for which only symptomatic treatment with limited benefits is available. AS is caused by mutations affecting the maternally inherited ubiquitin protein ligase E3A (UBE3A) gene. Previous studies showed that the silenced paternal Ube3a gene can be activated by targeting the antisense Ube3a-ATS transcript. We investigated antisense oligonucleotide–induced (ASO-induced) Ube3a-ATS degradation and its ability to induce UBE3A reinstatement and rescue of AS phenotypes in an established Ube3a mouse model. We found that a single intracerebroventricular injection of ASOs at postnatal day 1 (P1) or P21 in AS mice resulted in potent and specific UBE3A reinstatement in the brain, with levels up to 74% of WT levels in the cortex and a full rescue of sensitivity to audiogenic seizures. AS mice treated with ASO at P1 also showed rescue of established AS phenotypes, such as open field and forced swim test behaviors, and significant improvement on the reversed rotarod. Hippocampal plasticity of treated AS mice was comparable to WT but not significantly different from PBS-treated AS mice. No rescue was observed for the marble burying and nest building phenotypes. Our findings highlight the promise of ASO-mediated reactivation of UBE3A as a disease-modifying treatment for AS.

Normothermic machine perfusion (NMP) of donor kidneys provides the opportunity for improved graft preservation and objective pre-transplant ex-vivo organ assessment. Currently, a multitude of perfusion solutions exist for renal NMP. This study aimed to evaluate four different perfusion solutions side-by-side and determine the influence of different perfusate compositions on measured renal perfusion parameters. Porcine kidneys and blood were obtained from a slaughterhouse. Kidneys underwent NMP at 37°C for 7 hours, with 4 different perfusion solutions (n = 5 per group). Group 1 consisted of red blood cells (RBCs) and a perfusion solution based on Williams' Medium E. Group 2 consisted of RBCs, albumin and a balanced electrolyte composition. Group 3 contained RBCs and a medium based on a British clinical NMP solution. Group 4 contained RBCs and a medium used in 24-hour perfusion experiments. NMP flow patterns for solutions 1 and 2 were similar, solutions 3 and 4 showed lower but more stable flow rates. Thiobarbituric acid reactive substances were significantly higher in solution 1 and 4 compared to the other groups. Levels of injury marker N-acetyl-β-D glucosaminidase were significantly lower in solution 2 in comparison with solution 3 and 4. This study illustrates that the perfusate composition during NMP significantly impacts the measured perfusion and injury parameters and thus affects the interpretation of potential viability markers. Further research is required to investigate the individual influences of principal perfusate components to determine the most optimal conditions during NMP and eventually develop universal organ assessment criteria.

Glomerular endothelial cell (GEnC) dysfunction is important in the pathogenesis of glomerular sclerotic diseases, including Focal Segmental Glomerulosclerosis (FSGS) and overt diabetic nephropathy (DN). GEnCs form the first cellular barrier in direct contact with cells and factors circulating in the blood. Disturbances in these circulating factors can induce GEnC dysfunction. GEnC dysfunction occurs in early stages of FSGS and DN, and is characterized by a compromised endothelial glycocalyx, an inflammatory phenotype, mitochondrial damage and oxidative stress, aberrant cell signaling, and endothelial-to-mesenchymal transition (EndMT). GEnCs are in an interdependent relationship with podocytes and mesangial cells, which involves bidirectional cross-talk via intercellular signaling. Given that GEnC behavior directly influences podocyte function, it is conceivable that GEnC dysfunction may culminate in podocyte damage, proteinuria, subsequent mesangial activation, and ultimately glomerulosclerosis. Indeed, GEnC dysfunction is sufficient to cause podocyte injury, proteinuria and activation of mesangial cells. Aberrant gene expression patterns largely contribute to GEnC dysfunction and epigenetic changes seem to be involved in causing aberrant transcription. This review summarizes literature that uncovers the importance of cross-talk between GEnCs and podocytes, and GEnCs and mesangial cells in the context of the development of FSGS and DN, and the potential use of GEnCs as efficacious cellular target to pharmacologically halt development and progression of DN and FSGS.

IntroductionChimerism after orthotopic liver transplantation (OLT) has largely been investigated in intrahepatic cellular constituents. However, little is known about chimerism in the extrahepatic and large intrahepatic bile ducts. Our aim was to evaluate the presence and extent of chimerism after OLT in the peribiliary glands (PBG) and the luminal epithelium of the large donor bile ducts.MethodsFor this study, we examined six extrahepatic and large intrahepatic bile ducts from livers that were re-transplanted. In all cases there was a sex-mismatch between donor and recipient (female donor organ and male recipient), which allowed to discriminate between donor- and recipient-derived cells. Specimens from female to female transplants were used as negative controls and male to male transplants as positive controls. Fluorescence in situ hybridization (FISH) for Y and X chromosomes was performed and the percentage of XY positive cells was determined among biliary epithelial cells. Immunohistochemistry was used to correlate chimerism with histological features.ResultsCholangiocellular chimerism in all studied specimens ranged from 14 to 52%. The degree of chimerism was not associated with biliary damage. Marked chimerism was present at 5 days post-OLT. Ki-67-positivity was detected in 1–8% of the epithelial cells at the time of liver re-transplantation, and this correlated inversely with the degree of chimerism.ConclusionRecipient-derived cholangiocytes are present in the large bile ducts of the donor liver after OLT. The presence of chimerism in the large bile ducts suggests that recipient-derived cells may play a role in biliary regeneration following ischemia-induced injury during OLT.

In this work, a model of an energy system based on photovoltaics as the main energy source and a hybrid energy storage consisting of a short-term lithium-ion battery and hydrogen as the long-term storage facility is presented. The electrical and the heat energy circuits and resulting flows have been modelled. Therefore, the waste heat produced by the electrolyser and the fuel cell have been considered and a heat pump was considered to cover the residual heat demand. The model is designed for the analysis of a whole year energy flow by using a time series of loads, weather and heat profile as input. This paper provides the main set of equations to derive the component properties and describes the implementation into MATLAB/Simulink. The novel model was created for an energy flow simulation over one year. The results of the simulation have been verified by comparing them with well-established simulation results from HOMER Energy. It turns out that the novel model is well suited for the analysis of the dynamic system behaviour. Moreover, different characteristics to achieve an energy balance, an ideal dimensioning for the particular use case and further research possibilities of hydrogen use in the residential sector are covered by the novel model.

Abstract Purpose The histopathology of sepsis-associated acute kidney injury (AKI) in critically ill patients remains an understudied area. Previous studies have identified that acute tubular necrosis (ATN) is not the only driver of sepsis-AKI. The focus of this study was to identify additional candidate processes that may drive sepsis-AKI. To do this we immunohistochemically characterized the histopathological and cellular features in various compartments of human septic kidneys. Methods We studied the following histopathological features: leukocyte subsets, fibroblast activation, cellular proliferation, apoptosis, and fibrin deposition in the glomerulus and the tubulointerstitium in human post-mortem kidney biopsy tissue. Biopsy tissue samples from 27 patients with sepsis-AKI were collected 33 min (range 24–150) after death in the ICU. The unaffected part of the kidneys from 12 patients undergoing total nephrectomy as a result of renal carcinoma served as controls. Results Immunohistochemical analysis revealed the presence of more neutrophils and macrophages in the glomeruli and more neutrophils in the tubulointerstitium of renal tissue from patients with sepsis compared to control renal tissue. Type II macrophages were predominant, with some macrophages expressing both type I and type II markers. In contrast, there were almost no macrophages found in control kidneys. The number of activated (myo)fibroblasts was low in the glomeruli of sepsis-AKI kidneys, yet this was not observed in the tubulointerstitium. Cell proliferation and fibrin deposition were more pronounced in the glomeruli and tubulointerstitium of sepsis-AKI than in control kidneys. Conclusions The extensive heterogeneity of observations among and within patients emphasizes the need to thoroughly characterize patients with sepsis-AKI in a large sample of renal biopsy tissue from patients with sepsis.

Background and purpose — The number of revision total knee arthroplasties (TKA) is continuously increasing, leading to a growing need for reliable management of metaphyseal bone loss. We evaluated patients operated with a TKA using metal metaphyseal sleeves for bone defects with a minimum 5-year follow-up. Patients and methods — 37 patients had been operated on. 3 patients died and 3 patients were lost during follow-up. Of the 31 remainders (20 women), 9 had been operated on with a primary TKA and 22 with a revision TKA at the index surgery. The mean age at surgery was 69 (54–89) years and the mean follow-up time was 7.4 (5–12) years. Bone defects were classified according to the Anderson Orthopaedic Research Institute classification (tibia: type I n = 9, type II n = 5 and type III n = 17; femur: type I n = 12, type II n = 3 and type III n = 16). Results — At final follow-up one-third experienced an improvement concerning walking aids and walking distance. Except for 1 patient, all had full extension and a mean knee flexion of 110 (90–140) degrees. VAS pain at rest was 13 (SD 25) and on movement 30 (SD 31). 7 patients were reoperated due to: infection (n = 4), periprosthetic fracture (n = 1), skin necrosis (n = 1), and wound rupture (n = 1). The cumulative 5-year survival rate for reoperation was 77% (CI 63–92) and for revision 97% (CI 91–100). At the time of final follow-up, the sleeves showed good osseointegration with no signs of progressive radiolucency or migration. Interpretation — Titanium sleeves are a promising option in managing difficult cases with metaphyseal bone defects in TKA, providing a stable construct with good medium-term radiographic outcome

Background and purpose — Large metal-on-metal (MoM) articulations are associated with metal wear and corrosion, leading to increased metal ion concentrations and unacceptable revision rates. There are few comparative studies of 28-mm MoM articulations with conventional metal-on-polyethylene (MoP) couplings. We present a long-term follow-up of a randomized controlled trial comparing MoM versus MoP 28-mm articulations, focused on metal ions and implant survival. Patients and methods — 85 patients with a mean age of 65 years at surgery were randomized to a MoM (Metasul) or a MoP (Protasul) bearing. After 16 years, 38 patients had died and 4 had undergone revision surgery. 13 patients were unavailable for clinical follow-up, leaving 30 patients (n = 14 MoM and n = 16 MoP) for analysis of metal ion concentrations and clinical outcome. Results — 15-year implant survival was similar in both groups (MoM 96% [95% CI 88–100] versus MoP 97% [95% CI 91–100]). The mean serum cobalt concentration was 4-fold higher in the MoM (1.5 μg/L) compared with the MoP cohort (0.4 μg/L, p < 0.001) and the mean chromium concentration was double in the MoM (2.2 μg/L) compared with the MoP cohort (1.0 μg/L, p = 0.05). Mean creatinine levels were similar in both groups (MoM 93 μmol/L versus MoP 92 μmol/L). Harris hip scores differed only marginally between the MoM and MoP cohorts. Interpretation — This is the longest follow-up of a randomized trial on 28-mm MoM articulations, and although implant survival in the 2 groups was similar, metal ion concentrations remained elevated in the MoM cohort even in the long term.

Abstract PGC-1α is a versatile inducer of mitochondrial biogenesis and responsive to the changing energy demands of the cell. As mitochondrial ATP production requires proteins that derive from translation products of cytosolic ribosomes, we asked whether PGC-1α directly takes part in ribosomal biogenesis. Here, we show that a fraction of cellular PGC-1α localizes to the nucleolus, the site of ribosomal transcription by RNA polymerase I. Upon activation PGC-1α associates with the ribosomal DNA and boosts recruitment of RNA polymerase I and UBF to the rDNA promoter. This induces RNA polymerase I transcription under different stress conditions in cell culture and mouse models as well as in healthy humans and is impaired already in early stages of human Huntington’s disease. This novel molecular link between ribosomal and mitochondrial biogenesis helps to explain sarcopenia and cachexia in diseases of neurodegenerative origin.

Background: 2,4,5-Trimethoxyamphetamine (TMA-2) is a potent psychedelic compound. Structurally related 4-alkyloxy-substituted 2,5-dimethoxyamphetamines and phenethylamine congeners (2C-O derivatives) have been described but their pharmacology is mostly undefined. Therefore, we examined receptor binding and activation profiles of these derivatives at monoamine receptors and transporters.Methods: Receptor binding affinities were determined at the serotonergic 5-HT1A, 5-HT2A, and 5-HT2C receptors, trace amine-associated receptor 1 (TAAR1), adrenergic α1 and α2 receptors, dopaminergic D2 receptor, and at monoamine transporters, using target-transfected cells. Additionally, activation of 5-HT2A and 5-HT2B receptors and TAAR1 was determined. Furthermore, we assessed monoamine transporter inhibition.Results: Both the phenethylamine and amphetamine derivatives (Ki = 8–1700 nM and 61–4400 nM, respectively) bound with moderate to high affinities to the 5-HT2A receptor with preference over the 5-HT1A and 5-HT2C receptors (5-HT2A/5-HT1A = 1.4–333 and 5-HT2A/5-HT2C = 2.1–14, respectively). Extending the 4-alkoxy-group generally increased binding affinities at 5-HT2A and 5-HT2C receptors but showed mixed effects in terms of activation potency and efficacy at these receptors. Introduction of a terminal fluorine atom into the 4-ethoxy substituent by trend decreased, and with progressive fluorination increased affinities at the 5-HT2A and 5-HT2C receptors. Little or no effect was observed at the 5-HT1A receptor for any of the substances tested (Ki ≥ 2700 nM). Phenethylamines bound more strongly to the TAAR1 (Ki = 21–3300 nM) compared with their amphetamine analogs (Ki = 630–3100 nM).Conclusion: As seen with earlier series investigated, the 4-alkyloxy-substituted 2,5-dimethoxyamphetamines and phenethylamines share some trends with the many other phenethylamine pharmacophore containing compounds, such as when increasing the size of the 4-substituent and increasing the lipophilicity, the affinities at the 5-HT2A/C subtype also increase, and only weak 5-HT2A/C subtype selectivities were achieved. At least from the binding data available (i.e., high affinity binding at the 5-HT2A receptor) one may predict mainly psychedelic-like effects in humans, at least for some of the compound investigated herein.

Gladiatorial combats were much more than violent spectacles to entertain an enthusiastic audience. The first such events were held in connection to funerary practices and, as the Roman Empire grew, the combats gained a new dimension, this time related to the imperial cult. Moreover, the organisation, choreography and promotion of these combats become the embodiment of the new reality of the Roman world and its symbiosis with the Greek culture. This study aims to analyze one of the aspects of this symbiosis in the Black Sea, starting from the Greek and Latin terminology used in relation to the organisation of gladiatorial combats in the Pontic provinces.

Imaging modalities using tumor-directed monoclonal antibodies may be of value to improve the pre- and intraoperative detection and resection of lymph node (LN) metastatic disease in penile squamous cell carcinoma (PSCC). We investigated the expression of prostate-specific membrane antigen (PSMA), vascular endothelial growth factor (VEGF), epidermal growth factor receptor (EGFR) and epithelial cell adhesion molecule (EpCAM) to analyze their potency for diagnostic applications. Antigen expression was determined in primary tumors and LNs with and without metastases of 22 patients with PSCC. The total immunostaining score (TIS, 0–12) was determined as the product of a proportion score (PS, 0–4) and an intensity score (IS, 0–3). EGFR and VEGF expression were high in primary tumor (median TIS 8) and LN metastases (median TIS 6 and 8, respectively). No EGFR expression was seen in LNs without metastases. However, LNs without metastases did show VEGF expression (median TIS 6). No EpCAM or PSMA expression was seen in PSCC. This study shows that VEGF and EGFR expression is moderate to high in LN metastases of PSCC. Both VEGF and EGFR warrant further clinical evaluation to determine their value as a target for pre- and intraoperative imaging modalities in the detection of LN metastases in PSCC.

Sodium glucose cotransporter 2 inhibitors (SGLT2i) are the first antidiabetic compounds that effectively reduce heart failure hospitalization and cardiovascular death in type 2 diabetics. Being explicitly designed to inhibit SGLT2 in the kidney, SGLT2i have lately been investigated for their off-target cardiac actions. Here, we review the direct effects of SGLT2i Empagliflozin (Empa), Dapagliflozin (Dapa), and Canagliflozin (Cana) on various cardiac cell types and cardiac function, and how these may contribute to the cardiovascular benefits observed in large clinical trials. SGLT2i impaired the Na+/H+ exchanger 1 (NHE-1), reduced cytosolic [Ca2+] and [Na+] and increased mitochondrial [Ca2+] in healthy cardiomyocytes. Empa, one of the best studied SGLT2i, maintained cell viability and ATP content following hypoxia/reoxygenation in cardiomyocytes and endothelial cells. SGLT2i recovered vasoreactivity of hyperglycemic and TNF-α-stimulated aortic rings and of hyperglycemic endothelial cells. Anti-inflammatory actions of Cana in IL-1β-treated HUVEC and of Dapa in LPS-treated cardiofibroblast were mediated by AMPK activation. In isolated mouse hearts, Empa and Cana, but not Dapa, induced vasodilation. In ischemia-reperfusion studies of the isolated heart, Empa delayed contracture development during ischemia and increased mitochondrial respiration post-ischemia. Direct cardiac effects of SGLT2i target well-known drivers of diabetes and heart failure (elevated cardiac cytosolic [Ca2+] and [Na+], activated NHE-1, elevated inflammation, impaired vasorelaxation, and reduced AMPK activity). These cardiac effects may contribute to the large beneficial clinical effects of these antidiabetic drugs.

Individuals of many species rely on odors to communicate, find breeding partners, locate resources and sense dangers. In vertebrates, odorants are detected by chemosensory receptors of the olfactory system. One class of these receptors, the trace amine-associated receptors (TAARs), was recently suggested to mediate male sexual interest and mate choice. Here we tested this hypothesis in mice by generating a cluster deletion mouse (Taar2-9−/−) lacking all TAARs expressed in the olfactory epithelium, and evaluating transduction pathways from odorants to TAARs, neural activity and behaviors reflecting sexual interest. We found that a urinary volatile amine, isobutylamine (IBA), was a potent ligand for TAAR3 (but not TAAR1, 4, 5, and 6). When males were exposed to IBA, brain regions associated with sexual behaviors were less active in Taar2-9−/− than in wild type males. Accordingly, Taar2-9−/− males spent less time sniffing both the urine of females and pure IBA than wild type males. This is the first demonstration of a comprehensive transduction pathway linking odorants to TAARs and male sexual interest. Interestingly, the concentration of IBA in female urine varied across the estrus cycle with a peak during estrus. This variation in IBA concentration may represent a simple olfactory cue for males to recognize receptive females. Our results are consistent with the hypothesis that IBA and TAARs play an important role in the recognition of breeding partners and mate choice.

Trace amines (TAs), endogenous amino acid metabolites that are structurally similar to the biogenic amines, are endogenous ligands for trace amine-associated receptor 1 (TAAR1), a GPCR that modulates dopaminergic, serotonergic, and glutamatergic activity. Selective TAAR1 full and partial agonists exhibit similar pro-cognitive, antidepressant- and antipsychotic-like properties in rodents and non-human primates, suggesting TAAR1 as a novel target for the treatment of neurological and psychiatric disorders. We previously reported that TAAR1 partial agonists are wake-promoting in rats and mice, and that TAAR1 knockout (KO) and overexpressing mice exhibit altered sleep-wake and EEG spectral composition. Here, we report that locomotor and EEG spectral responses to the psychostimulants modafinil and caffeine are attenuated in TAAR1 KO mice. TAAR1 KO mice and WT littermates were instrumented for EEG and EMG recording and implanted with telemetry transmitters for monitoring locomotor activity (LMA) and core body temperature (Tb). Following recovery, mice were administered modafinil (25, 50, 100 mg/kg), caffeine (2.5, 10, 20 mg/kg) or vehicle p.o. at ZT6 in balanced order. In WT mice, both modafinil and caffeine dose-dependently increased LMA for up to 6 h following dosing, whereas only the highest dose of each drug increased LMA in KO mice, and did so for less time after dosing. This effect was particularly pronounced following caffeine, such that total LMA response was significantly attenuated in KO mice compared to WT at all doses of caffeine and did not differ from Vehicle treatment. Tb increased comparably in both genotypes in a dose-dependent manner. TAAR1 deletion was associated with reduced wake consolidation following both drugs, but total time in wakefulness did not differ between KO and WT mice. Furthermore, gamma band EEG activity following both modafinil and caffeine treatment was attenuated in TAAR1 KO compared to WT mice. Our results show that TAAR1 is a critical component of the behavioral and cortical arousal associated with two widely used psychostimulants with very different mechanisms of action. Together with our previous findings, these data suggest that TAAR1 is a previously unrecognized component of an endogenous wake-modulating system.

Objective: Type 2 diabetes and obesity are emerging pandemics in the 21st century creating worldwide urgency for the development of novel and safe therapies. We investigated trace amine-associated receptor 1 (TAAR1) as a novel target contributing to the control of glucose homeostasis and body weight. Methods: We investigated the peripheral human tissue distribution of TAAR1 by immunohistochemistry and tested the effect of a small molecule TAAR1 agonist on insulin secretion in vitro using INS1E cells and human islets and on glucose tolerance in C57Bl6, and db/db mice. Body weight effects were investigated in obese DIO mice. Results: TAAR1 activation by a selective small molecule agonist increased glucose-dependent insulin secretion in INS1E cells and human islets and elevated plasma PYY and GLP-1 levels in mice. In diabetic db/db mice, the TAAR1 agonist normalized glucose excursion during an oral glucose tolerance test. Sub-chronic treatment of diet-induced obese (DIO) mice with the TAAR1 agonist resulted in reduced food intake and body weight. Furthermore insulin sensitivity was improved and plasma triglyceride levels and liver triglyceride content were lower than in controls. Conclusions: We have identified TAAR1 as a novel integrator of metabolic control, which acts on gastrointestinal and pancreatic islet hormone secretion. Thus TAAR1 qualifies as a novel and promising target for the treatment of type 2 diabetes and obesity.

Lightweight boards and beams in the wood-based construction and furniture industry are not a new topic. The density reduction of panels using sandwich structure with light cores was confirmed by users like doors or mobile homes more than three decades ago. Today many ways to attain a lighter wooden structure are on offer, partially in industrial application. The first one is the use of light-weight wood species like balsa, lime, pine from southern hemisphere plantations etc. limited by the availability, strength properties, gluability and so on. A second one is the sandwich structure made from hard faces like thick veneer, thin plywood, particleboard or high density thin fiberboard and cores made from honeycomb paper, very light wood species or foams like the polystyrene one. A third way to produce a light structure is to reduce the core drastically, using predesigned skeletons with special shapes and connections to the faces. The engines for these developments are on the one hand the fast growing market of knockdown furniture and on the other hand the increasing costs for energy and raw materials. Additional factors that make weight saving a primary economical objective for most producers are transportation costs, easier handling and higher acceptance among the end users. Moreover, customers demand more for ergonomical solutions regarding packaging. Many patents were generated by researchers and developers for new one-stage production processes for sandwich panels with wood- and impregnated paper-based facings made from veneers, particles or fibres and a core consisting of expandable foams, particles or embedded hard skeletons. These ideas or prototypes could be integrated in existing continuous pressing lines for wood based panels keeping some of the advantages of the continuous production technique in matters of efficiency. Some of the challenges of the light weight wooden structure are the connection in half or final parts, resistance to water, moisture, temperature and fire, and maybe last but not least the recyclability due to the mixture of different materials like foams, plastics etc.

Due to the ever-increasing scarcity of resources and raw materials in the wood panels industry, it is imperative to look for suitable alternatives to the established resources. Therefore a combination of the traditionally used and newly explored sources may reveal highly innovative ways. The objective of this study is to provide an insight into the behavior of the material and possible new applications of those fiber/particle wood and waste leather composites. For this reason exclusively fibers of spruce were used for the trials. Wet white (WW) leather particles and wet blue (WB) leather particles were mixed with the wooden materials for the production of high density fibreboards. Besides the mechanical properties such as the internal bond (IB) the bending strength (MOR) and modulus of elasticity (MOE) was analyzed. Further physical property as thickness swelling after 24h watering was investigated. To analyze how the density influences the behavior under thermal conditions, fiberboards with the densities 500, 700 and 900 kg/m³ were tested. The results of the material properties were influenced by the leather content of the panels. The results for the UF-bonded HDF boards show enhancement of the transverse IB with increasing wet blue leather content, whereas the other mechanical properties decline meanwhile. The thickness swelling showed higher values compared to the wood fibreboard. The results of this study underline the usefulness of integrating leather shavings to HDF and give an overview of their influence in wood fiber materials. The combination of the natural resource wood fiber and the leather waste products (Wet Blue and Wet White) gives a very interesting new material, its mechanical properties allow a variety of possible application in future applications.

The tendency in recent decades for manufacturing plants of semi-finished products such as composite panels, has been to invest in order to achieve high production capacities (>2,000 m³/day for panels and >3,000 t/day for paper with one line). The trend of concentrating the primary processing capacities and manufacturing wood-based panels will continue for the next few years not only in Europe but in North and South America as well. The ten largest panel manufacturers had a combined manufacturing capacity that exceeded a third of the worldwide production capacity. The financial crisis that started in 2008 has caused the closure of a large number of factories especially in North America and Central Europe. Small- and medium-sized producers will only survive if they will continue to specialize in the manufacture of panel types and sizes (niche products) that are “unprofitable” for mega-groups. The installed production capacity worldwide of all wood-based composite panels combined (includes PY, PB, MDF, OSB) rose by more than 2.5 times between 1980 and 2005 (225 mil.m³), and continues to increase despite the crises reaching approx. 300 mil.m³ in 2013. The forecast for the coming years varies greatly from continent to continent. In North America and Central Europe, both a consolidation of the available production capacities and the closure of less efficient older lines are expected. The lowest point of the effect of the financial crisis on the building industry seems to have been overcome. The furniture production companies will continue to move from one continent and region to another.

Do verbally reported feelings follow reinforcement learning principles? Prediction errors—differences between expectations and outcomes—are key in models of learning across humans, animals, and machines. Historically, the emphasis has been on outcomes in the environment (e.g., money or food), focusing relatively less on the fact that humans can also report correspondingly expected and experienced affect (i.e., feelings). Recent research suggests that expected and experienced affect, including prediction errors, can explain behavior beyond outcomes in the environment alone. However, the properties of affective prediction errors underlying this explanatory power are unknown. We address this gap across two studies. We show that affective prediction errors can decrease over time, but that the decrease depends on introspection (Study 1). We then replicate this finding while additionally documenting transfer effects across tasks (Study 2). Crucially, decreases in affective prediction errors generally occurred independent of changes in behavior.

To prevent renal graft thrombosis in kidney transplantation, centres use different perioperative anticoagulant strategies, based on various risk factors. In our centre, patients transplanted preemptively are considered at increased risk of renal graft thrombosis compared to patients who are dialysis-dependent at time of transplantation. Therefore these patients are given a single dose of 5000 IU unfractionated heparin intraoperatively before clamping of the vessels. We questioned whether there is a difference in haemostatic state between preemptively and non-preemptively transplanted patients and whether the distinction in intraoperative heparin administration used in our center is justified. For this analysis, citrate samples of patients participating in the VAPOR-1 trial were used and several haemostatic and fibrinolytic parameters were measured in 29 preemptively and 28 non-preemptively transplanted patients and compared to 37 living kidney donors. Sample points were: induction anaesthesia (T1), 5 minutes after reperfusion (T2) and 2 hours postoperative (T3). At T1, recipient groups showed comparable elevated levels of platelet factor 4 (PF4, indicating platelet activation), prothrombin fragment F1+2 and D-dimer (indicating coagulation activation) and Von Willebrand Factor (indicating endothelial activation) compared to the donors. The Clot Lysis Time (CLT, a measure of fibrinolytic potential) was prolonged in both recipient groups compared to the donors. At T3, F1+2, PF4 and CLT were higher in non-preemptively transplanted recipients compared to preemptively transplanted recipients. Compared to donors, non-preemptive recipients showed a prolonged CLT, but comparable levels of PF4 and D-dimer. In conclusion pre-transplantation, preemptively and non-preemptively transplanted patients show a comparable enhanced haemostatic state. A distinction in intraoperative heparin administration between preemptive and non-preemptive transplantation does not seem justified.

Studies on microtubule inhibitors have shown that the loss of equilibrium between tubulin and microtubules can generate a multitude of histochemical changes in mitochondria. This disruption of balance is also considered the basis of benzimidazole anthelmintic (BZ) activity. Studies have shown that BZ does not bond to the tubulin of the BZ-resistant Haemonchus contortus, as opposed to sensitive ones. This affinity alteration can be easily recognised by changes in the optical density and can help in the classification of H. contortus mitochondria, into sensitive (dark) and resistant (clear, unmodified zones). In order to confirm this hypothesis, we started our study from albendazole (ABZ) resistant and sensible H. contortus individuals, collected from the intestinal tract of sheep, aiming towards the identification of mitochondrial features, using the Electron Microscopy Transmission (EMT) technique. The EMT has confirmed that the structure of sensitive trichostrongilian populations was affected rapidly, only four hours after ABZ treatments. The main changes that appeared in the intestinal mitochondria of sensitive helminths were: cristae thickening and decreasing in number and cellular membrane thickening. Twelve hours after anthelmintic administration, a total blocking of metabolic functionality was observed, and finally, these changes completely altered the optical density of the mitochondria. In ABZ resistant populations, the optical density has remained normal; and the cristae number, size or functionality of resistant nematode mitochondria has remained unchanged.

Background: Project success used to be measured solely in terms of efficiency metrics such as scope, cost and time; however, there are proposals that more attention should be paid to process-related performance factors such as communication. The advent of email has significantly impacted the way the world communicates. Objectives: This study investigates the preference of email communication relative to other communication mediums in project environments and the effect of email communication on feelings of stress and overload in the workplace. Method: A survey with 430 responses was conducted to determine the communication preferences of project practitioners in a typical project. The average rank and frequency response methods were used to analyse the data. Results: The findings indicate that the communication preferences of project practitioners still support the media richness theory and that face-to-face communication is the preferred communication medium in most situations. Despite email being disruptive and a cause of stress, the respondents did not indicate being overloaded because of email. Conclusion: Even though there has been a dramatic shift towards email and electronic communication in projects, face-to-face communication is still the most preferred communication type for most situations. Furthermore, email is perceived as an effective tool to delegate, can be used to build and develop relationships and trust, and is an efficient and effective tool that contributes to project communication success.

The ultra large multi-agent systems are becoming increasingly popular due to quick decay of the individual production costs and the potential of speeding up the solving of complex problems. Examples include nano-robots, or systems of nano-satellites for dangerous meteorite detection, or cultures of stem cells for organ regeneration or nerve repair. The topics associated with these systems are usually dealt within the theories of intelligent swarms or biologically inspired computation systems. Stochastic models play an important role and they are based on various formulations of the mechanical statistics. In these cases, the main assumption is that the swarm elements have a simple behaviour and that some average properties can be deduced for the entire swarm. In contrast, complex systems in areas like aeronautics are formed by elements with sophisticated behaviour, which are even autonomous. In situations like this, a new approach to swarm coordination is necessary. We present a stochastic model where the swarm elements are communicating autonomous systems, the coordination is separated from the component autonomous activity and the entire swarm can be abstracted away as a piecewise deterministic Markov process, which constitutes one of the most popular model in stochastic control. Keywords: ultra large multi-agent systems, system of systems, autonomous systems, stochastic hybrid systems.

Phytoplasmas are cell wall-less plant pathogenic bacteria responsible for major crop losses throughout the world. In grapevine they cause grapevine yellows, a detrimental disease associated with a variety of symptoms. The high economic impact of this disease has sparked considerable interest among researchers to understand molecular mechanisms related to pathogenesis. Increasing evidence exist that a class of small non-coding endogenous RNAs, known as microRNAs (miRNAs), play an important role in post-transcriptional gene regulation during plant development and responses to biotic and abiotic stresses. Thus, we aimed to dissect complex high-throughput small RNA sequencing data for the genome-wide identification of known and novel differentially expressed miRNAs, using read libraries constructed from healthy and phytoplasma-infected Chardonnay leaf material. Furthermore, we utilised computational resources to predict putative miRNA targets to explore the involvement of possible pathogen response pathways. We identified multiple known miRNA sequence variants (isomiRs), likely generated through post-transcriptional modifications. Sequences of 13 known, canonical miRNAs were shown to be differentially expressed. A total of 175 novel miRNA precursor sequences, each derived from a unique genomic location, were predicted, of which 23 were differentially expressed. A homology search revealed that some of these novel miRNAs shared high sequence similarity with conserved miRNAs from other plant species, as well as known grapevine miRNAs. The relative expression of randomly selected known and novel miRNAs was determined with real-time RT-qPCR analysis, thereby validating the trend of expression seen in the normalised small RNA sequencing read count data. Among the putative miRNA targets, we identified genes involved in plant morphology, hormone signalling, nutrient homeostasis, as well as plant stress. Our results may assist in understanding the role that miRNA pathways play during plant pathogenesis, and may be crucial in understanding disease symptom development in aster yellows phytoplasma-infected grapevines.

Background: Testing for HER2 amplification and/or overexpression is currently routine practice to guide Herceptin therapy in invasive breast cancer. At present, HER2 status is most commonly assessed by immunohistochemistry (IHC). Standardization of HER2 IHC assays is of utmost clinical and economical importance. At present, HER2 IHC is most commonly performed with the HercepTest which contains a polyclonal antibody and applies a manual staining procedure. Analytical variability in HER2 IHC testing could be diminished by a fully automatic staining system with a monoclonal antibody.