Background: as we look to extend ex vivo lung perfusion times (EVLP) to improve preservation, the metabolic activity of the lungs will require support from other organ functions. Active functional liver support, including detoxification, synthesis, and regulation, can improve lung preservation during EVLP. This study aimed to demonstrate the effects of hepatic conditioning of the EVLP perfusate on lung endothelium, via the receptor of advanced glycation end-products (RAGE)-nuclear-factor-κB (NF-κB) signaling in vitro. Methods: we performed in vitro experiments using human lung microvascular endothelial cells (HLMVECs), human hepatocytes, and perfusate (Steen solution). Four experimental groups: 1) fresh Steen (negative controls, NC), 2) EVLP’ed Steen control, this solution collected after 12 h of EVLP of human lungs, 3) hepatocyte conditioned EVLP’ed Steen (Hep-cond.), and 4) a RAGE inhibitor added in EVLP’ed Steen (RAGE inhibitor). HLMVECs were incubated in each testing condition and exposed to hypoxia (1% O2/8% CO2) for 24 h. Media were collected to investigate NF-κB signaling and endothelial glycocalyx damage. Results: HLMVECs incubated under hypoxia in EVLP’ed Steen showed significantly upregulated NF-κB signal and endothelial damage denoted by increased glycosaminoglycans and matrix metalloproteinase-2 activity among the groups. The Hep-cond. solution significantly attenuated those findings, while the RAGE inhibitor attenuated the NF-κB signal but not endothelial glycocalyx damage. Conclusion: Our study demonstrates that hepatic function incorporated into EVLP can ameliorate pulmonary endothelial cells injury under hypoxic normothermic perfusion exposure. Our data supports the concept of incorporating other organ functions into an organ perfusion platform, to enhance lung graft preservation.
Hamzah Rehan, Jack A Maggiore, Anastasia L. Gant Kanegusuku
OBJECTIVE The OrganOx metra is a recent innovation approved by the US Food and Drug Administration that enables continuous ex vivo perfusion of a donor liver prior to transplantation. A mixture of human blood products, added nutrients, and drugs is perfused throughout the liver to sustain its viability. This study determines whether the perfusate matrix affects the analytical accuracy of biochemical parameters used to evaluate the function of the donor liver before transplantation. METHODS A mixing study was conducted to evaluate the percent recovery of total bilirubin, aspartate aminotransferase, alanine aminotransferase, lactic acid, glucose, and lactate dehydrogenase in OrganOx metra perfusate using the Beckman Coulter AU5800 automated platform. Perfusate pools were mixed with pooled patient plasma (Li Hep) in the following ratios: 1:4, 1:1, and 4:1. These mixtures were measured alongside straight patient plasma and straight perfusate. RESULTS Agreement between expected and measured values, with a percent recovery for all tested analytes, was between 92% and 109%. Detrimental matrix effects were not observed. CONCLUSIONS The Beckman Coulter AU5800 analyzer provides reliable and accurate biochemical analysis of OrganOx metra perfusate during ex vivo preservation. The findings validate its suitability for perfusate analysis, supporting its potential use in clinical and research settings.
Bioengineered livers are currently an acceptable alternative to orthotopic liver transplants to overcome the scarcity of donors. However, the challenge of using a bioengineered liver is the lack of an intact endothelial layer in the vascular network leading to thrombosis. Heparin-modified surfaces have been demonstrated to decrease thrombogenicity in earlier research. However, in our study, we aimed to apply heparin immobilization to enhance the hemocompatibility, endothelial cell (EC) adhesion, and angiogenesis of rat decellularized liver scaffolds (DLS). Heparin was immobilized on the DLS by the end-point attachment technique. The scaffold’s hemocompatibility was assessed using ex vivo blood perfusion and platelet adhesion studies. The heparinized scaffold (HEP-DLS) showed a significantly reduced thrombogenicity and platelet aggregation. HEP-DLS was recellularized with EA.hy926 cells via the portal vein and maintained in the bioreactor for 7 days, showing increased EC adhesion and coverage within the blood vessels. The Resazurin reduction assay confirmed the presence of actively proliferating cells in the HEP-DLS. The scaffolds were implanted subcutaneously into the dorsum of mice for 21 days to evaluate cell migration and angiogenesis. The results showed significant increases in the number of blood vessels in the HEP-DLS group. Our results demonstrated that heparin immobilization reduces thrombosis, promotes re-endothelialization, and enhances angiogenesis in DLS. The research provides insight into the potential use of heparin in the formation of a functioning vasculature.
Background and Aims Precise diagnostic biomarkers are urgently required for pancreatic ductal adenocarcinoma (PDAC). Therefore, the aim of this study was to identify PDAC-specific exosomal microRNAs (Ex-miRs) from pancreatic juice (PJ) and evaluate their diagnostic potential. Methods Exosomes in PJ and serum were extracted using ultracentrifugation and confirmed morphologically and biochemically. PDAC-specific Ex-miRs were identified using our original miR arrays, in which “Ex-miRs derived from the PJ of patients with chronic pancreatitis (CP)” were subtracted from Ex-miRs commonly expressed in both “human PDAC cell lines” and “the PJ of patients with PDAC.” We verified the expression of these miRs using quantitative real-time reverse transcription polymerase chain reaction. Changes in serum Ex-miR levels were assessed in 2 patients with PDAC who underwent curative resection. In situ hybridization was performed to directly visualize PDAC-specific miR expression in cancer cells. Results We identified novel Ex-miR-4516 and Ex-miR-4674 from the PJ of patients with PDAC, and they showed 80.0% and 81.8% sensitivity, 80.8% and 73.3% specificity, and 90.9% and 80.8% accuracy, respectively. The sensitivity, specificity, and accuracy of a triple assay of Ex-miR-4516/4674/PJ cytology increased to 93.3%, 81.8%, and 88.5%, respectively. In serum samples (n = 88), the sensitivity, specificity, and accuracy of Ex-miR-4516 were 97.5%, 34.3%, and 68%, respectively. Presurgical levels of serum-derived Ex-miR-4516 in 2 patients with relatively early disease stages declined after curative resection. In situ hybridization demonstrated that Ex-miR-4516 expression exclusively occurred in cancer cells. Conclusion Liquid assays using the in situ-proven Ex-miR-4516 may have a high potential for detecting relatively early-stage PDAC and monitoring its clinical course.
It remains a challenge to endow a polymeric material with antithrombotic ability by surface grafting without disturbing the bulk properties of the substrate. Heparin-based functional structures of less than 80 nm were fabricated and covalently grafted on a polyethylene terephthalate surface via carbene chemistry (Hep-g-PET). Heparin was oxidized with the minimum antithrombrin sequence retained, creating an aldehyde group on the chain terminus. Oxidized heparin was then covalently attached to a poly(amidoamine) (PAMAM)-grafted PET substrate. The interface between blood and PET was improved by the surface functionality, and the amount of attached platelets decreased to 29 ± 12.1% of its initial value. The bulk properties of the functionalized film were hardly influenced, and the visible light transmittance remained more than 96%. The tethered structures also showed the ability to kill attached S. aureus and E. coli efficiently. The functionalized membrane showed negligible ex vivo cell cytotoxicity and a low hemolysis ratio. Hep-g-PET was implanted in between rat skin and muscle, and showed an outstanding histological response and antimicrobial ability. The influences of the graft thickness and the heparin chain length were explored. The strategies reported in this work may help to improve the design of polymeric implant bio-devices.
The mini proceedings of the "Fourth International Workshop on the Extension Project for the J-PARC Hadron Experimental Facility (HEF-ex 2024) [https://kds.kek.jp/event/46965]" held at J-PARC, February 19-21, 2024, are presented. The workshop was devoted to discussing the physics case that connects both the present and the future Hadron Experimental Facility at J-PARC, covering a wide range of topics in flavor, hadron, and nuclear physics related to both experimental and theoretical activities being conducted at the facility.
Maximilian Horzela, Henri Casanova, Manuel Giffels
et al.
Predicting the performance of various infrastructure design options in complex federated infrastructures with computing sites distributed over a wide area network that support a plethora of users and workflows, such as the Worldwide LHC Computing Grid (WLCG), is not trivial. Due to the complexity and size of these infrastructures, it is not feasible to deploy experimental test-beds at large scales merely for the purpose of comparing and evaluating alternate designs. An alternative is to study the behaviours of these systems using simulation. This approach has been used successfully in the past to identify efficient and practical infrastructure designs for High Energy Physics (HEP). A prominent example is the Monarc simulation framework, which was used to study the initial structure of the WLCG. New simulation capabilities are needed to simulate large-scale heterogeneous computing systems with complex networks, data access and caching patterns. A modern tool to simulate HEP workloads that execute on distributed computing infrastructures based on the SimGrid and WRENCH simulation frameworks is outlined. Studies of its accuracy and scalability are presented using HEP as a case-study. Hypothetical adjustments to prevailing computing architectures in HEP are studied providing insights into the dynamics of a part of the WLCG and candidates for improvements.
The Caltech HEP Crystal Lab has been actively investigating novel inorganic scintillators along the following three directions. Fast and radiation hard inorganic scintillators to face the challenge of severe radiation environment expected by future HEP experiments at hadron colliders, such as the high luminosity LHC and FCC hh. Ultrafast inorganic scintillators to face the challenge of unprecedented event rate expected by future HEP experiments searching for rare decays, such as Mu2e II, and ultrafast time of flight system at hadron colliders. Cost effective inorganic scintillators for the homogeneous hadron calorimeter concept to face the challenge of both electromagnetic and jet mass resolutions required by the proposed Higgs factory. We report novel materials along all directions: LuAG:Ce ceramic fibers for the HL LHC, Lu2O3:Yb ceramic scintillators for ultrafast applications, and ABS:Ce and DSB:Ce glass scintillators for the proposed Higgs factory. The result of this investigation may also benefit nuclear physics experiments, GHz hard X ray imaging, medical imaging, and homeland security applications.
The shortcomings of the Standard Model (SM) motivate its extension to accommodate new expected phenomena, such as dark matter and neutrino masses. However, such extensions are generally more complex due to the presence of a large number of free parameters and additional phenomenology. Understanding how theoretical and experimental limits affect the parameter spaces of new models, individually and collectively, is of utmost importance for conducting model status analysis, motivating precise computations, or model-building aimed at solving certain issues. However, checking the constraints usually require a large amount of time using a chain of physics tools. We demonstrate, for the first time, the application of deep learning (DL) for the multilabel classification (MLC) of a group of theoretical and experimental constraints in the dark doublet phase of the next-to-two-Higgs-doublet model (DDP-N2HDM), as a representative 9-dimensional parameter space. We analyze the issue of class imbalance and the ability of the classifier to learn joint class distributions. We demonstrate the time advantage compared to physics tools, with the classifier achieving orders of magnitude faster checks on groups of constraints and strong performance. The classifier performed strongly in terms of identifying regions where all constraints are valid or invalid, as well as regions where one or more of the constraints are valid or invalid simultaneously. This approach can be applied to any extension beyond the SM with the potential to aid HEP tools or act as a surrogate for fast model status checks. To that end, we provide a python tool \texttt{HEPMLC} for generating and investigating multilabel classifiers for SM extensions.
We present a brief report (at the Nufact-2024 conference) summarizing a global extraction of the ${\rm ^{12}C}$ longitudinal (${\cal R}_L$) and transverse (${\cal R}_T$) nuclear electromagnetic response functions from an analysis of all available electron scattering data on carbon. Since the extracted response functions cover a large kinematic range they can be readily used for comparison to theoretical predictions as well as validation and tuning Monte Carlo (MC) generators for electron and neutrino scattering experiments. Comparisons to several theoretical approaches and MC generators are given in arXiv:2409.10637v1 [hep-ex]. We find that among all the theoretical models that were investigated, the ``Energy Dependent-Relativistic Mean Field'' (ED-RMF) approach provides the best description of both the Quasielastic (QE) and {\it nuclear excitation} response functions (leading to single nucleon final states) over all values of four-momentum transfer. he QE data are also well described by the"Short Time Approximation Quantum Monte Carlo"(STA-QMC) calculation which includes both single and two nucleon final states which presently is only valid for momentum transfer $0.3$ 0.65 GeV requires the implementation of relativistic corrections. Both approaches have the added benefit that the calculations are also directly applicable to the same kinematic regions for neutrino scattering. In addition we also report on a universal fit to all electron scattering data that can be used in lieu of experimental data for validation of Monte Carlo generators (and is in the process of being implemented in GENIE).
Emerging evidence indicates that a vicious cycle between inflammation and microthrombosis catalyzes the pathogenesis of inflammatory bowel disease (IBD). Over‐stimulated inflammation triggers a coagulation cascade and leads to microthrombosis, which further complicates the injury through tissue hypoxia and ischemia. Herein, an injectable protein hydrogel with anti‐thrombosis and anti‐inflammation competency is developed to impede this cycle, cross‐linked by silver ion mediated metal‐ligand coordination and electronic interaction with sulfhydryl functionalized bovine serum albumin and heparin, respectively. The ex vivo experiments show that the hydrogel, HEP‐Ag‐BSA, exhibits excellent self‐healing ability, injectability, biocompatibility, and sustained drug release. HEP‐Ag‐BSA also demonstrates anti‐coagulation and anti‐inflammation abilities via coagulation analysis and lipopolysaccharide stimulation assay. The in vivo imaging confirms the longer retention time of HEP‐Ag‐BSA at inflammatory sites than in normal mucosa owing to electrostatic interactions. The in vivo study applying a mouse model with colitis also reveals that HEP‐Ag‐BSA can robustly inhibit inflammatory microthrombosis with reduced bleeding risk. This versatile protein hydrogel platform can definitively hinder the “inflammation and microthrombosis” cycle, providing a novel integrated approach against IBD.
The presence of microplastics in commercially important seafood species is a new issue of food safety concern. Although plastic debris has been found in the gastrointestinal tracts of several species, the prevalence of microplastics in edible shrimp tissues in Thailand has not yet been established. For the first time, the gastrointestinal tract (GT), heptapancreas (HEP), muscle (MU) and exoskeleton (EX) of farmed white leg shrimp (Litopenaeus vannamei) from commercial aquaculture facilities in Nakhon Pathom Province, Thailand, were analyzed for microplastics (MPs). The number of MP items per tissue was 27.36±2.28 in the GT, 17.42±0.90 in the HEP, 11.37±0.60 in the MU and 10.04±0.52 in the EX. MP concentrations were 137.78±16.48, 16.31±1.87, 1.69±0.13 and 4.37±0.27 items/gram (ww) in the GT, HEP, MU and EX, respectively. Microplastics ranged in size from < 100 to 200–250 μm, with fragment-shape (62.07%), fibers (37.31%) and blue (43.69%) was the most common. The most frequently found polymers in shrimp tissue organs and pond water were polyethylene terephthalate (PET), polyvinyl acetate (PVAc) and cellulose acetate butyrate (CAB). Shrimp consumption (excluding GT and EX) was calculated as 28.79 items/shrimp/person/day using Thailand's consumption of shrimp, MP abundance and shrimp consumption. The results of the study can be used as background data for future biomonitoring of microplastics in shrimp species that are significant from an ecological and commercial perspective. MP abundance in farmed L. vannamei may be related to feeding habits and the source of MPs could come from the aquaculture facilities operations.