Heart-beating Donors Research Articles

Recent solid-phase microextraction-based analytical approaches for the profiling of biliary bile acids in pre-transplant assessments of liver grafts subjected to normothermic ex vivo liver perfusion

BackgroundLiver transplantation is the definitive treatment for end-stage liver failure, but the scarcity of donor organs remains a significant challenge. Leveraging organs from extended criteria donors (ECD) offers a potential avenue to address worldwide shortages, though these organs are more susceptible to post-reperfusion injury. This study explores the use of normothermic ex vivo liver perfusion (NEVLP) as a method for organ preservation – an approach that sustains liver metabolism and facilitates pre-transplant assessments of organ viability via bile analysis. The focal point of this study revolves on the development of analytical methods for determining the bile acid profile throughout the peritransplantation period as a potential indicator of liver function and viability. ResultsThe study optimized and validated a high-throughput analytical method to quantify selected bile acids in bile samples using a thin-film microextraction-liquid chromatography-mass spectrometry (TFME-LC-MS) platform. Furthermore, it introduced a solid-phase microextraction-microfluidic open interface-mass spectrometry (SPME-MOI-MS) method for rapid direct analysis of bile acid isobar groups. In the animal study, discernible variations in the concentrations of specific bile acids were observed between donors after circulatory death (DCD) and heart-beating donors (HBD), particularly following normothermic perfusion and reperfusion. Noteworthy fluctuations in individual bile acid concentrations were observed throughout the entire organ transplantation process, with taurocholic acid (TCA), glycocholic acid (GCA), and glycochenodeoxycholic acid (GCDCA) emerging as promising indicators of organ quality. The efficacy of the SPME-MOI-MS platform in corroborating these trends highlights its potential for real-time bile acid analysis during liver transplantation procedures. SignificanceOur findings underscore the efficacy of NEVLP in tandem with advanced bile acid analysis methods as a reliable strategy for pre-transplant assessments of organ viability, potentially increasing the use of ECD organs and reducing organ shortages. The ability to monitor bile acid profiles in real-time provides crucial insights into liver function and ischemic injury, making significant strides in improving transplant outcomes and patient survival rates.

Loss of insulin-expressing extra-islet cells in type 1 diabetes is accompanied with increased number of glucagon-expressing extra-islet cells.

The presence of remaining insulin-positive cells in type 1 diabetes (T1D) is well-known. These cells are part of islets or appear as extra-islet insulin-positive cells scattered in the exocrine parenchyma. The latter are poorly described, and the presence of scattered endocrine cells expressing other islet hormones than insulin has not been explored. This study aimed to compare the extra-islet insulin- or glucagon-positive cells concerning their frequency, transcription-factor expression, and mitotic activity in subjects with and without T1D. Multispectral imaging was used to examine extra-islet cells by staining for insulin, glucagon, ARX, PDX1, and Ki67. This was done in well-preserved pancreatic tissue obtained from heart-beating organ donors with or without T1D. In three T1D donors, lobes with insulin-containing islets (ICI) were found. Within these, a higher frequency of extra-islet insulin-positive cells was observed compared to lobes with insulin-deficient islets (IDI). Increased frequency of glucagon-positive extra-islet cells was observed in donors with T1D (median 53 cells/mm2) when compared with non-diabetic donors (11 cells/mm2, p = 0.004). Proliferating endocrine cells were present in donors with, and without T1D, as demonstrated by Ki67-positive staining (0-3% of the cells expressing insulin or glucagon). The reduced frequency of extra-islet insulin-positive cells in lobes with IDI in donors with T1D suggests that the pathological mechanism causing beta cell demise in T1D affects entire lobes. The presence of an increased frequency of glucagon-positive extra-islet cells supports the notion of a preserved capacity to regenerate the endocrine pancreas in donors with T1D.

Indocyanine green fluorescence quantification during normothermic ex situ perfusion for the assessment of porcine liver grafts after circulatory death.

Current graft evaluation during normothermic ex situ liver perfusion lacks real-time parameters for predicting posttransplant hepatocyte and biliary function. Indocyanine green (ICG) imaging has been widely used in liver surgery, enabling the visualization of hepatic uptake and excretion through bile using near-infrared light. In this research, porcine livers under various ischemic conditions were examined during a 5-hour normothermic ex situ liver perfusion procedure, introducing ICG at 1 hour through the hepatic artery. These conditions included livers from heart-beating donors, donation after circulatory death (DCD) with warm ischemic durations of 60 minutes (DCD60) and 120 minutes (DCD120), as well as interventions utilizing tissue plasminogen activator in DCD120 cases (each n = 5). Distinct hepatic fluorescence patterns correlated with different degrees of ischemic injury ( p = 0.01). Low ICG uptake in the parenchyma (less than 40% of maximum intensity) was more prevalent in DCD120 (21.4%) compared to heart-beating donors (6.2%, p = 0.06) and DCD60 (3.0%, p = 0.02). Moreover, ICG clearance from 60 minutes to 240 minutes was significantly higher in heart-beating donors (69.3%) than in DCD60 (17.5%, p < 0.001) and DCD120 (32.1%, p = 0.01). Furthermore, thrombolytic intervention using tissue plasminogen activator in DCD120 resulted in noteworthy outcomes, including significantly reduced ALP levels ( p = 0.04) and improved ICG clearance ( p = 0.02) with a trend toward mitigating fibrin deposition similar to DCD60, as well as enhancements in bile production ( p = 0.09). In conclusion, ICG fluorescence imaging during normothermic ex situ liver perfusion provides real-time classification of hepatic vascular and biliary injuries, offering valuable insights for the more accurate selection and postintervention evaluation of marginal livers in transplantation.

Metabolomic and lipidomic landscape of porcine kidney associated with kidney perfusion in heart beating donors and donors after cardiac death

Transplant centers are currently facing a lack of tools to ensure adequate evaluation of the quality of the available organs, as well as a significant shortage of kidney donors. Therefore, efforts are being made to facilitate the effective use of available organs and expand the donor pool, particularly with expanded criteria donors. Fulfilling a need, we aim to present an innovative analytical method based on solid-phase microextraction (SPME) - chemical biopsy. In order to track changes affecting the organ throughout the entire transplant procedure, porcine kidneys were subjected to multiple samplings at various time points. The application of small-diameter SPME probes assured the minimal invasiveness of the procedure. Porcine model kidney autotransplantation was executed for the purpose of simulating two types of donor scenarios: donors with a beating heart (HBD) and donors after cardiac death (DCD). All renal grafts were exposed to continuous normothermic ex vivo perfusion. Following metabolomic and lipidomic profiling using high-performance liquid chromatography coupled to a mass spectrometer, we observed differences in the profiles of HBD and DCD kidneys. The alterations were predominantly related to energy and glucose metabolism, and differences in the levels of essential amino acids, purine nucleosides, lysophosphocholines, phosphoethanolamines, and triacylglycerols were noticed. Our results indicate the potential of implementing chemical biopsy in the evaluation of graft quality and monitoring of renal function during perfusion.

Axonal mapping of motor and sensory components within the ulnar nerve and its branches.

Intrinsic function is indispensable for dexterous hand movements. Distal ulnar nerve defects can result in intrinsic muscle dysfunction and sensory deficits. Although the ulnar nerve's fascicular anatomy has been extensively studied, quantitative and topographic data on motor axons traveling within this nerve remain elusive. The ulnar nerves of 14 heart-beating organ donors were evaluated. The motor branches to the flexor carpi ulnaris (FCU) and flexor digitorum profundus (FDP) muscles and the dorsal branch (DoBUN) as well as 3 segments of the ulnar nerve were harvested in 2-cm increments. Samples were subjected to double immunofluorescence staining using antibodies against choline acetyltransferase and neurofilament. Samples revealed more than 25,000 axons in the ulnar nerve at the forearm level, with a motor axon proportion of only 5%. The superficial and DoBUN showed high axon numbers of more than 21,000 and 9300, respectively. The axonal mapping of more than 1300 motor axons revealed an increasing motor/sensory ratio from the proximal ulnar nerve (1:20) to the deep branch of the ulnar nerve (1:7). The motor branches (FDP and FCU) showed that sensory axons outnumber motor axons by a ratio of 10:1. Knowledge of the detailed axonal architecture of the motor and sensory components of the human ulnar nerve is of the utmost importance for surgeons considering fascicular grafting or nerve transfer surgery. The low number of efferent axons in motor branches of the ulnar nerve and their distinct topographical distribution along the distal course of the nerve is indispensable information for modern nerve surgery.

Robotic Bladder Autotransplantation: Preclinical Studies in Preparation for First-in-human Bladder Transplant.

Human urinary bladder transplantation has never been performed. From a technical standpoint, challenges include the complex deep pelvic vascular anatomy, limited intraoperative visualization, and high procedural complexity. In preparation for a first-in-human clinical trial, we report preclinical studies to develop the technique of robotic retrieval and autotransplantation of vascularized composite bladder allograft. Institutional Animal Care and Use Committee, Institutional Review Board, and UNOS (United Network for Organ Sharing) approvals were obtained, and IDEAL (Idea, Development, Exploration, Assessment, Long-term Study) Reporting Guidelines were followed. Robotic vascularized composite bladder allograft recovery, back-table graft preparation, and robotic autotransplantation were performed in 3 vascularized model settings: living porcine (n=3), pulsatile human cadavers (n=2), and heart-beating brain-dead deceased research human donors (n=5). Our primary objective was to develop a reproducible technique for robotic vascularized composite bladder allograft transplantation. Technical success was defined by adequate, sustained vascularized composite bladder allograft reperfusion. Secondary objectives were intraoperative parameters, including operative time, graft ischemia time, and blood loss. Successful robotic vascularized composite bladder allograft autotransplantation was achieved in 2 porcine, 1 cadaver, and 3 brain-dead research donors. In the heart-beating research donors, console time decreased with successive surgeries, and visual inspection revealed healthy revascularized autografts with prompt, global indocyanine green immunofluorescence uptake. In 1 heart-beating donor who was hemodynamically maintained for 12 hours postoperatively, reinspection confirmed excellent maintained global vascularized composite bladder allograft vascularity and bladder mucosal integrity. To our knowledge, the first preclinical experience of bladder autotransplantation in vascularized models is reported, including robotic vascularized composite bladder allograft recovery, back-table reconstruction, and autotransplantation. This experience represents the essential preclinical work required to build toward the first-in-human trial of bladder transplantation, performed under a UNOS-approved genitourinary vascularized composite bladder allograft program (NCT No. 05462561).

Pancreatic Islet Viability Assessment Using Hyperspectral Imaging of Autofluorescence.

Islets prepared for transplantation into type 1 diabetes patients are exposed to compromising intrinsic and extrinsic factors that contribute to early graft failure, necessitating repeated islet infusions for clinical insulin independence. A lack of reliable pre-transplant measures to determine islet viability severely limits the success of islet transplantation and will limit future beta cell replacement strategies. We applied hyperspectral fluorescent microscopy to determine whether we could non-invasively detect islet damage induced by oxidative stress, hypoxia, cytokine injury, and warm ischaemia, and so predict transplant outcomes in a mouse model. In assessing islet spectral signals for NAD(P)H, flavins, collagen-I, and cytochrome-C in intact islets, we distinguished islets compromised by oxidative stress (ROS) (AUC = 1.00), hypoxia (AUC = 0.69), cytokine exposure (AUC = 0.94), and warm ischaemia (AUC = 0.94) compared to islets harvested from pristine anaesthetised heart-beating mouse donors. Significantly, with unsupervised assessment we defined an autofluorescent score for ischaemic islets that accurately predicted the restoration of glucose control in diabetic recipients following transplantation. Similar results were obtained for islet single cell suspensions, suggesting translational utility in the context of emerging beta cell replacement strategies. These data show that the pre-transplant hyperspectral imaging of islet autofluorescence has promise for predicting islet viability and transplant success.

Infarct Size With Incremental Global Myocardial Ischemia Times: Cyclosporine A in Donation After Circulatory Death Rat Hearts

We quantified the myocardial infarct size with varying global ischemia durations and studied the benefits of Cyclosporine A (CyA) in reducing cardiac injury in ex vivo and transplanted rat hearts. Infarct size was measured after 15, 20, 25, 30, and 35 minutes of in vivo global ischemia (n=34) and compared with control beating-heart donor (CBD) hearts (n=10). For heart function assessment, donation after circulatory death (DCD) rat hearts (n=20) were procured after 25 minutes of in vivo ischemia and reanimated ex vivo for 90 minutes. Half of the DCD hearts received CyA (0.5 mM) at reanimation. The CBD hearts (n=10) served as controls. A separate group of CBD and DCD (with or without CyA treatment) hearts underwent heterotopic heart transplantation; heart function was measured at 48 hours. Infarct size was 25% with 25 minutes of ischemia and increased significantly with 30 and 35 minutes to 32% and 41%, respectively. CyA treatment decreased infarct size in DCD hearts (15% vs 25%). Heart function in the transplanted DCD hearts was significantly better with CyA treatment and was comparable to CBD hearts. CyA administered at reperfusion limited infarct size in DCD hearts and improved their function in transplanted hearts.

Islet transplantation: overcoming the organ shortage

BackgroundType 1 diabetes mellitus (T1D) is a condition resulting from autoimmune destruction of pancreatic β cells, leading patients to require lifelong insulin therapy, which, most often, does not avoid the most common complications of this disease. Transplantation of isolated pancreatic islets from heart-beating organ donors is a promising alternative treatment for T1D, however, this approach is severely limited by the shortage of pancreata maintained under adequate conditions.MethodsIn order to analyze whether and how this problem could be overcome, we undertook a retrospective study from January 2007 to January 2010, evaluating the profile of brain-dead human pancreas donors offered to our Cell and Molecular Therapy NUCEL Center (www.usp.br/nucel) and the basis for organ refusal.ResultsDuring this time period, 558 pancreata were offered by the São Paulo State Transplantation Central, 512 of which were refused and 46 were accepted for islet isolation and transplantation. Due to the elevated number of refused organs, we decided to analyze the main reasons for refusal in order to evaluate the possibility of improving the organ acceptance rate. The data indicate that hyperglycemia, technical issues, age, positive serology and hyperamylasemia are the top five main causes for declination of a pancreas offer.ConclusionsThis study underlines the main reasons to decline a pancreas offer in Sao Paulo—Brazil and provides some guidance to ameliorate the rate of eligible pancreas donors, aiming at improving the islet isolation and transplantation outcome.Trial registration: Protocol CAPPesq number 0742/02/CONEP 9230.

V08-01 ROBOTIC BLADDER AUTOTRANSPLANTATION IN A BRAIN-DEAD HEART-BEATING DECEASED DONOR: WET RUN BEFORE A FIRST-IN-HUMAN CLINICAL TRIAL

You have accessJournal of UrologyCME1 Apr 2023V08-01 ROBOTIC BLADDER AUTOTRANSPLANTATION IN A BRAIN-DEAD HEART-BEATING DECEASED DONOR: WET RUN BEFORE A FIRST-IN-HUMAN CLINICAL TRIAL Nima Nassiri, Giovanni Cacciamani, David Ginsberg, Evgeniy Kreydin, Andre Abreu, Mihir Desai, Monish Aron, Yasir Qazi, Yuri Genyk, and Inderbir Gill Nima NassiriNima Nassiri More articles by this author , Giovanni CacciamaniGiovanni Cacciamani More articles by this author , David GinsbergDavid Ginsberg More articles by this author , Evgeniy KreydinEvgeniy Kreydin More articles by this author , Andre AbreuAndre Abreu More articles by this author , Mihir DesaiMihir Desai More articles by this author , Monish AronMonish Aron More articles by this author , Yasir QaziYasir Qazi More articles by this author , Yuri GenykYuri Genyk More articles by this author , and Inderbir GillInderbir Gill More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003306.01AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Bladder transplantation has not yet been successfully performed. Technical challenges include complicated deep pelvic vascular anatomy, limited visualization, and procedural complexity. In preparation for a first-in-human clinical trial of bladder transplantation (NCT05462561), we performed a robotic vascularized composite bladder allograft (VCBA) autotransplantation in a brain-dead heart-beating research donor. METHODS: An appropriate research donor was identified by our local organ procurement organization. VCBA autotransplantation was performed, including VCBA dissection, extracorporeal back-tabling, vascular reanastomoses, and vesicourethral anastomosis. A robotic approach was necessary to optimize visualization in the deep pelvis and maximally preserve allograft perfusion. The primary objective was to develop the stepwise procedure and gauge its technical success, defined by autograft perfusion by direct visual inspection and indigo-cyanine green (ICG) fluorescence. Secondary objectives included intraoperative parameters (operative times, warm ischemia time, and EBL). IRB (HS-21-00162) approval was obtained. RESULTS: Technical success, defined by adequate autograft perfusion after vascular anastomosis, was achieved. In the brain-dead heart beating donor, the technical maneuvers were well tolerated, with intra-operative hemodynamic stability (Table 1). Operative time was 670 min. Visual inspection revealed a pink, healthy bladder and patent vascular anastomoses, with diffuse ICG uptake. CONCLUSIONS: This is the first documented case of robotic bladder autotransplantation, performed in a brain-dead heart-beating research donor as a necessary preclinical step towards a first-in-human trial of bladder transplantation (NCT05462561). Technical feasibility was demonstrated, and vascular viability was confirmed by ICG uptake. The robotic approach was necessary for fine vascular dissection in the deep pelvis. Source of Funding: This study has been supported by a clinical research grant from the OneLegacy Foundation. © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e748 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Nima Nassiri More articles by this author Giovanni Cacciamani More articles by this author David Ginsberg More articles by this author Evgeniy Kreydin More articles by this author Andre Abreu More articles by this author Mihir Desai More articles by this author Monish Aron More articles by this author Yasir Qazi More articles by this author Yuri Genyk More articles by this author Inderbir Gill More articles by this author Expand All Advertisement PDF downloadLoading ...

MP65-05 ROBOTIC BLADDER AUTOTRANSPLANTATION: PRECLINICAL STUDIES IN PREPARATION FOR FIRST-IN-HUMAN TRIAL OF BLADDER TRANSPLANTATION

You have accessJournal of UrologyCME1 Apr 2023MP65-05 ROBOTIC BLADDER AUTOTRANSPLANTATION: PRECLINICAL STUDIES IN PREPARATION FOR FIRST-IN-HUMAN TRIAL OF BLADDER TRANSPLANTATION Nima Nassiri, Giovanni Cacciamani, David Ginsberg, Evgeniy Kreydin, Andre Abreu, Mihir Desai, Monish Aron, Yasir Qazi, Yuri Genyk, and Inderbir Gill Nima NassiriNima Nassiri More articles by this author , Giovanni CacciamaniGiovanni Cacciamani More articles by this author , David GinsbergDavid Ginsberg More articles by this author , Evgeniy KreydinEvgeniy Kreydin More articles by this author , Andre AbreuAndre Abreu More articles by this author , Mihir DesaiMihir Desai More articles by this author , Monish AronMonish Aron More articles by this author , Yasir QaziYasir Qazi More articles by this author , Yuri GenykYuri Genyk More articles by this author , and Inderbir GillInderbir Gill More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003323.05AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Bladder transplantation has not yet been successfully performed. Technical challenges include complicated deep pelvic vascular anatomy, limited visualization, and procedural complexity. In preparation for a first-in-human clinical trial of bladder transplantation (NCT05462561), we describe our pre-clinical experience with robotic vascularized composite bladder allograft (VCBA) autotransplantation. METHODS: VCBA autotransplantation was performed in three settings: living porcine, pulsatile human cadaver, and brain-dead heart-beating deceased research donor. A robotic approach was utilized to optimize visualization in the deep pelvis and maximally preserve allograft perfusion. The primary objective was development of the stepwise procedure and gauging its technical success, defined by autograft perfusion by direct visual inspection and indigo-cyanine green (ICG) fluorescence. Secondary objectives included intraoperative parameters (operative times, warm ischemia time, and EBL). IACUC (21277) and IRB (HS-21-00162) approval were obtained. The IDEAL Reporting Guidelines were followed. RESULTS: VCBA procurement and autotransplantation was performed in 3 pigs, 2 pulsatile-perfused cadavers, and 1 brain-dead, heart-beating deceased donor (Table 1). Technical success was achieved in two porcine, one cadaveric, and the brain dead deceased donor cases. Operative times in the porcine models ranged from 143-343 min, improving sequentially with each case. A similar finding was noted in the cadaveric model. In the brain-dead heart beating donor, the surgery was well tolerated, with intra-operative hemodynamic stability. Operative time was 670 min. Visual inspection revealed a pink, healthy bladder and patent vascular anastomoses, with diffuse ICG uptake. CONCLUSIONS: Porcine and cadaveric models were used to develop a protocol for VCBA procurement and transplantation. This protocol was applied to the first brain-dead heart-beating donor, documenting technically feasibility. The robotic approach facilitates the fine vascular dissection in the deep pelvis. The above cases are necessary preclinical steps towards a first-in-human trial of bladder transplantation. Source of Funding: This study has been supported by a clinical research grant from the OneLegacy Foundation. © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e892 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Nima Nassiri More articles by this author Giovanni Cacciamani More articles by this author David Ginsberg More articles by this author Evgeniy Kreydin More articles by this author Andre Abreu More articles by this author Mihir Desai More articles by this author Monish Aron More articles by this author Yasir Qazi More articles by this author Yuri Genyk More articles by this author Inderbir Gill More articles by this author Expand All Advertisement PDF downloadLoading ...

Electron Transport Chain Inhibition to Decrease Injury in Transplanted Donation After Circulatory Death Rat Hearts.

Donation after circulatory death (DCD) donor hearts sustain ischemic damage and are not routinely used for heart transplantation. DCD heart injury, particularly reperfusion injury, is primarily mediated by releasing reactive oxygen species from the damaged mitochondria (complex I of the electron transport chain). Amobarbital (AMO) is a transient inhibitor of complex I and is known to reduce releasing reactive oxygen species generation. We studied the beneficial effects of AMO in transplanted DCD hearts. Sprague-Dawley rats were assigned to 4 groups-DCD or DCD + AMO donors and control beating-heart donors (CBD) or CBD + AMO donors (n = 6-8 each). Anesthetized rats were connected to a ventilator. The right carotid artery was cannulated, heparin and vecuronium were administered. The DCD process started by disconnecting the ventilator. DCD hearts were procured after 25 minutes of in-vivo ischemia, whereas CBD hearts were procured without ischemia. At procurement, all donor hearts received 10 mL of University of Wisconsin cardioplegia solution. The CBD + AMO and DCD + AMO groups received AMO (2 mM) dissolved in cardioplegia. Heterotopic heart transplantation was performed by anastomosing the donor aorta and pulmonary artery to the recipient's abdominal aorta and inferior vena cava. After 14 days, transplanted heart function was measured with a balloon tip catheter placed in the left ventricle. Compared with CBD hearts, DCD hearts had significantly lower developed pressure. AMO treatment significantly improved cardiac function in DCD hearts. Treatment of DCD hearts at the time of reperfusion with AMO resulted in an improvement of transplanted heart function that was comparable with the CBD hearts.

Organ procurement requiring the authorization of the judicial authority. An epidemiological investigation in the Veneto region (Italy)

BackgroundWhen death occurs through a mechanism requiring a forensic investigation, consent for organ harvesting must be sought from the Judicial Authority (JA). AimTo perform a retrospective study of potential organ donors in the Veneto region over a six-year period (2012–2017), analysing any differences between cases in which the JA approved or denied organ harvesting. Material and methodsBoth non-heart beating (NHB) and heart beating (HB) donors were included. For HB cases, personal and clinical data were collected. To evaluate the correlation between the JA response and the circumstantial and clinical data a logistic multivariate analysis was performed, estimating the adjusted odds ratios (adjORs). ResultsBetween 2012 and 2017, 17,662 organ and/or tissue donors were included, of which 16,418 were NHB donors and 1,244 HB-donors. Among the 1,244 HB-donors, JA authorization was asked in 200 cases (16.1%), approved in 154 cases (77.0%), limited in 7 cases (3.5%) and denied in 39 cases (19.5%). The JA denied the authorization for organ harvesting in 53,3% of cases with hospitalizations of less than 1 day and in 9,4% of cases with hospitalization exceeding one week [adjOR(95%CI) = 10.67 (1.92–59.22)]. The performance of an autopsy was linked to a higher chance of denied outcome from the JA [adjOR(95%CI): 3.45 (1.42–8.39)]. ConclusionsImprovements in the communication between organ procurement organizations and the JA through efficient protocols furnishing detailed information on the cause of death might lead to a better procurement process with an increase in the number of transplanted organs.

Biliary Metabolome Profiling for Evaluation of Liver Metabolism and Biliary Tract Function Related to Organ Preservation Method and Degree of Ischemia in a Porcine Model.

The development of surgical techniques, immunosuppressive strategies and new organ preservation methods have meant that transplant centers have to face the problem of an insufficient number of organs for transplantation concerning the constantly growing demand. Therefore, using organs from expanded criteria donors and developing new analytical solutions to find parameters or compounds that would allow a more efficient assessment of organ quality before transplantation are options for meeting this challenge. This study proposed bile metabolomic analysis to evaluate liver metabolism and biliary tract function depending on the organ preservation method and degree of warm ischemia time. The analyses were performed on solid-phase microextraction-prepared bile samples from porcine model donors with mild (heart beating donor [HBD]) and moderate warm ischemia (donation after circulatory death [DCD]) grafts subjected to static cold storage (SCS) or normothermic ex vivo liver perfusion (NEVLP) before transplantation. Bile produced in the SCS-preserved livers was characterized by increased levels of metabolites such as chenodeoxycholic acid, arachidonic acid and 5S-hydroxyeicosatetraeonic acid, as well as saturated and monounsaturated lysophosphatidylcholines (LPC). Such changes may be associated with differences in the bile acid synthesis pathways and organ inflammation. Moreover, it has been shown that NEVLP reduced the negative effect of ischemia on organ function. A linear relationship was observed between levels of lipids from the LPC group and the time of organ ischemia. This study identified metabolites worth considering as potential markers of changes occurring in preserved grafts.

Tissue distribution of ethanol after intraprostatic injection using a porous needle.

To develop a safe and precise method for intraprostatic injection, and to establish correlation between the volume of ethanol injectate and the volume of subsequent infiltrated prostate tissue. We performed intraprostatic injection of 96% ethanol using a needle which has a segment of its wall made of capillary membrane with hundreds of pores in an acute and chronic canine experiment, in heart-beating cadaveric organ donors, and in a xenograft model of human prostate cancer. Whole mount tissue sections were used for three-dimensional reconstruction of the necrotic lesions and calculation of their volumes. The ethanol injection resulted in oval shaped lesions of well-delineated coagulative necrosis. In both healthy human and canine prostates, the prostatic pseudocapsule and neurovascular bundle remained intact without evidence of disruption. There was a linear correlation between administered volume of ethanol and the volume of necrotic lesion. Regression analysis showed strong correlation in the acute canine experiments and in experiments performed on xenografts of human prostate cancer. A formula was calculated for each experiment to estimate the relationship between the injected volume and the volume of infiltrated prostate tissue area. Intraprostatic injection using a porous needle allows for effective and predictable tissue distribution of the injectate in the prostate. Through varying the volume of the agent injected and use of needles with a different length of the porous segment, the volume of infiltrated tissue could be adjusted allowing for targeted focal treatment.

The ischaemic preconditioning paradox and its implications for islet isolation from heart-beating and non heart-beating donors

The impact of ischaemia can severely damage procured donor organs for transplantation. The pancreas, and pancreatic islets in particular, is one of the most sensitive tissues towards hypoxia. The present study was aimed to assess the effect of hypoxic preconditioning (HP) performed ex-vivo in islets isolated from heart-beating donor (HBD) and non heart-beating donor (NHBD) rats. After HP purified islets were cultured for 24 h in hypoxia followed by islet characterisation. Post-culture islet yields were significantly lower in sham-treated NHBD than in HBD. This difference was reduced when NHBD islets were preconditioned. Similar results were observed regarding viability, apoptosis and in vitro function. Reactive oxygen species generation after hypoxic culture was significantly enhanced in sham-treated NHBD than in HBD islets. Again, this difference could be diminished through HP. qRT-PCR revealed that HP decreases pro-apoptotic genes but increases HIF-1 and VEGF. However, the extent of reduction and augmentation was always substantially higher in preconditioned NHBD than in HBD islets. Our findings indicate a lower benefit of HBD islets from HP than NHBD islets. The ischaemic preconditioning paradox suggests that HP should be primarily applied to islets from marginal donors. This observation needs evaluation in human islets.

Abstract 12689: Electron Transport Chain Inhibition to Decrease Reperfusion Injury in Transplanted Donation After Circulatory Death Rat Hearts

Introduction: Intrinsic to the donation after circulatory death (DCD) donation is ischemia and reperfusion injury, primarily mediated through release of reactive oxygen species (ROS) from the damaged mitochondria. Amobarbital (AMO) is a transient, reversible inhibitor of complex I known to decrease ROS generation. We studied whether AMO treatment during reperfusion can improve function in transplanted DCD hearts. Methods: Sprague Dawley rats, were assigned to four heart donor groups- DCD or DCD+AMO and control beating-heart donor (CBD) or CBD+AMO (n=6-8 each). Donor rats were anesthetized using ketamine/xylazine, intubated and connected to a ventilator. The carotid access was used to deliver heparin (1000 U/kg) and vecuronium bromide (4 mg/kg) to paralyze diaphragmatic muscles. The DCD process was initiated by disconnecting the ventilator. Donor hearts were procured following 25 minutes of in-vivo ischemia in the DCD groups, while CBD hearts were procured without terminating the ventilation. At procurement, after clamping the aortic arch, all donor hearts received 10 ml of University of Wisconsin cardioplegia solution delivered through the carotid cannula. The CBD+AMO and DCD+AMO groups received AMO (2 mM), dissolved in the cardioplegia. Heterotopic heart transplantation was performed in recipient rat’s abdomen. After 14 days the transplanted heart function was measured with a balloon tip catheter. Results: Compared to CBD hearts, DCD hearts had significantly lower developed pressure, the rate pressure product (developed pressure times heart rate), and negative dP/dt indicating decreased contractile and impaired diastolic function (Figure). AMO treatment of the DCD hearts resulted in significantly improved cardiac function compared to the untreated DCD hearts. Conclusions: A single treatment with AMO during the heart procurement improves the transplanted DCD heart function which was comparable to the control CBD hearts.

Development of Non heart-beating donor programs for kidney transplantation in Italy: A perspective view

Kidney transplantation is the best treatment available for patients with end-stage renal disease. There are three categories of donors: living donors (LD), brain death (DBD) or cardiac death (DCD) kidney donors. In this review we analyze the current scenario of DCD renal transplantation in Europe and in Italy. In Italy DCD donation has not yet been significantly developed because of the obstacle of the 20 minutes no touch period after cardiac arrest, before cannulating and perfusing organs to be recovered. This procedure is requested by current Italian law and is not going to be changed in the near future. We analyze problems rising from this law and suggest possible solutions.

313.1: Use of Transient Liver Elastography in the Evaluation of Liver Viability for Organ Donation and Transplantation

Introduction: The lack of enough livers for transplant worldwide is a reality and in México because of donation rates and high prevalence of morbid obesity this condition is worse. Strategies to determine whether a liver is suitable for donation depends on many occasions on the predonation estimation of fatty content. For March 2022, exist 244 patients in waiting list for liver transplant. In 2021 only 135 transplants were performed out of 293 brain death donors (none non heart beating donors). The reasons why 53.9% of the livers are not use are wide but one of them is because the higher prevalence of overweight and obesity in Mexican population, meaning a high number of steatosis of the liver. Since there is also a poor air connectivity among Mexican cities, the risk for primary organ failure is increased by longer ischemia time. Because the common study to evaluate liver steatosis is ultrasound despite the poor sensitivity, we proposed to use Transient elastography as a bedside tool to evaluate fat content or fibrosis of the liver condition before harvesting, minimizing the mobilization of complex and expensive logistics necessary to harvest a liver just to be discarded later and assuring better transplant results. Method: A prospective double-blind study was conducted in Hospital General del Estado from October 2015 to November 2021 that consist in that in every donor, no matter body mass index, a liver ultrasound was made by a radiologist and later a liver elastography was made on the same donor by another radiologist. Then the results were confronted during the organ harvesting by surgeon expertise who decided to take liver or not. Results: We had 32 brain death organ donors to whom an ultrasound and liver elastography was made. Median age was 41 years and weight were 74.66 kgs, with a range from 55 to 120 kgs. By using conventional ultrasound, out of the 32 donors, 93.8% were diagnosed without liver steatosis and 3.1% with mild and 3.1% with severe, while elastography showed 50% no steatosis, 15.6 light, 6.3 mild and 28.1 severe. Correlation of Transient elastography with surgical appearance was 100 % (No biopsy was taken to correlate this results with pathology).Conclusion: Our study showed that using ultrasound for evaluate liver steatosis for organ donation is not as good as it might be thought. For no liver steatosis concordance between ultrasound and elastography was 50% but in cases for severe infiltration, diagnosed rate was 1 Vs 9 for ultrasound vs elastography, and severe steatosis confirmed at the moment of surgery being evident the superiority of elastography for diagnosis of mild to severe steatosis and becoming a better resource in the evaluation of brain death organ donor.

Machine Perfusion of the Human Heart

This brief communication about machine perfusion of potential human donor hearts describes its historical development. Included in the review are both the isolated perfusion of donor hearts retrieved from heart beating and non-heart-beating donors. Additionally, some detail of in-situ (within the donor body) normothermic regional reperfusion of the heart and other organs is given. This only applies to the DCD donor heart. Similarly, some detail of ex-situ (outside the body) heart perfusion is offered. This article covers the entire history of the reperfusion of donor hearts. It takes us up to the current day describing 6 years follow-up of these donor machine perfused hearts. These clinical results appear similar to the outcomes of heart beating donors if reperfusion is managed within 30 min of normothermic circulatory determined death. Future developments are also offered. These are 3-fold and include: i. the pressing need for objective markers of the clinical outcome after transplantation, ii. the wish for isolated heart perfusion leading to improvement in donor heart quality, and iii. a strategy to safely lengthen the duration of isolated heart perfusion.