Standard Donor Research Articles

Extracorporeal immune cell therapy of sepsis: ex vivo results

BackgroundImmune cell dysfunction plays a central role in sepsis-associated immune paralysis. The transfusion of healthy donor immune cells, i.e., granulocyte concentrates (GC) potentially induces tissue damage via local effects of neutrophils. Initial clinical trials using standard donor GC in a strictly extracorporeal bioreactor system for treatment of septic shock patients already provided evidence for beneficial effects with fewer side effects, by separating patient and donor immune cells using plasma filters. In this ex vivo study, we demonstrate the functional characteristics of a simplified extracorporeal therapy system using purified granulocyte preparations.MethodsPurified GC were used in an immune cell perfusion model prefilled with human donor plasma simulating a 6-h treatment. The extracorporeal circuit consisted of a blood circuit and a plasma circuit with 3 plasma filters (PF). PF1 is separating the plasma from the patient’s blood. Plasma is then perfused through PF2 containing donor immune cells and used in a dead-end mode. The filtrated plasma is finally retransfused to the blood circuit. PF3 is included in the plasma backflow as a redundant safety measure. The donor immune cells are retained in the extracorporeal system and discarded after treatment. Phagocytosis activity, oxidative burst and cell viability as well as cytokine release and metabolic parameters of purified GCs were assessed.ResultsCells were viable throughout the study period and exhibited well-preserved functionality and efficient metabolic activity. Course of lactate dehydrogenase and free hemoglobin concentration yielded no indication of cell impairment. The capability of the cells to secret various cytokines was preserved. Of particular interest is equivalence in performance of the cells on day 1 and day 3, demonstrating the sustained shelf life and performance of the immune cells in the purified GCs.ConclusionResults demonstrate the suitability of a simplified extracorporeal system. Furthermore, granulocytes remain viable and highly active during a 6-h treatment even after storage for 3 days supporting the treatment of septic patients with this system in advanced clinical trials.

Effect of Sperm Selection by Magnetic-Activated Cell Sorting in D-IUI: A Randomized Control Trial.

Clinical outcome in assisted reproduction techniques (ARTs) is mainly influenced by the quality of gametes used. It is known that a high percentage of sperm DNA fragmentation (DNAf) decreases the success of ART clinical results. Therefore, techniques such as magnetic-activated cell sorting (MACS) help to improve results in cases of patients with a high percentage of DNAf. Cryopreservation of sperm in donor intrauterine insemination (D-IUI) treatments increases sperm DNAf, so patients using these sperm samples can benefit from using this technique. This prospective randomized national multicenter study analyzed clinical outcomes of 181 D-IUI treatments. MACS was performed after density gradient centrifugation (DGC) in 90 thawed semen donor samples (MACSG), whereas only DGC was performed in 91 thawed semen donor samples (CG). To our knowledge, this is the first study analyzing the effect of MACS on D-IUI cycles. Our results show no significant differences in gestation, live birth, or miscarriage rates between the two groups. We believe that further studies with a larger sample size are needed to evaluate the application of MACS in combination with standard IUI donor sperm preparations in fertility clinics.

Marginal versus Standard Donors in Heart Transplantation: Proper Selection Means Heart Transplant Benefit.

Background: In this study, we assessed the mid-term outcomes of patients who received a heart donation from a marginal donor (MD), and compared them with those who received an organ from a standard donor (SD). Methods: All patients who underwent HTx between January 2012 and December 2020 were enrolled at a single institution. The primary endpoints were early and long-term survival of MD recipients. Risk factors for primary graft failure (PGF) and mortality in MD recipients were also analyzed. The secondary endpoint was the comparison of survival of MD versus SD recipients. Results: In total, 238 patients underwent HTx, 64 (26.9%) of whom received an organ from an MD. Hospital mortality in the MD recipient cohort was 23%, with an estimated 1 and 5-year survival of 70% (59.2–82.7) and 68.1% (57.1–81), respectively. A multivariate analysis in MD recipients showed that decreased renal function and increased inotropic support of recipients were associated with higher mortality (p = 0.04 and p = 0.03). Cold ischemic time (p = 0.03) and increased donor inotropic support (p = 0.04) were independent risk factors for PGF. Overall survival was higher in SD than MD (85% vs. 68% at 5 years, log-rank = 0.008). However, risk-adjusted mortality (p = 0.2) and 5-year conditional survival (log-rank = 0.6) were comparable. Conclusions: Selected MDs are a valuable resource for expanding the cardiac donor pool, showing promising results. The use of MDs after prolonged ischemic times, increased inotropic support of the MD or the recipient and decreased renal function are associated with worse outcomes.

MP36-04 THE “CANNIBALIZATION” OF ALTRUISTIC KIDNEY DONATION BY VOUCHER-BASED KIDNEY TRANSPLANTATION

You have accessJournal of UrologyCME1 May 2022MP36-04 THE “CANNIBALIZATION” OF ALTRUISTIC KIDNEY DONATION BY VOUCHER-BASED KIDNEY TRANSPLANTATION Nima Nassiri, Torey Averick, and Jeffrey Veale Nima NassiriNima Nassiri More articles by this author , Torey AverickTorey Averick More articles by this author , and Jeffrey VealeJeffrey Veale More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002590.04AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Barriers to living kidney donation are multi-factorial, and may include logistic, financial, familial, cultural, and religious factors. Less discussed but impactful barriers involve: (1) a reluctance to donate because a present-day donation prohibits future one, and (2) chronological incompatibility. Voucher-based kidney donation seeks to address these latter two barriers. Herein, we sought to evaluate how voucher-based donations have impacted standard altruistic kidney donation. METHODS: Subjects were all patients participating in standard altruistic donation and voucher-based donation through the National Kidney Registry from 2014 to present. Demographic and clinical factors were considered. The impact on downstream chains of transplantation were also evaluated. Both voucher-based and standard altruistic donation were considered “non-directed”. RESULTS: Since 2014, the total number of non-directed donations, either from standard altruistic donation or through the voucher program, has risen by over 3-fold (Figure 1). The number standard altruistic donations has declined since 2018, coinciding with an uptick in the number of voucher-based donations from 22 in 2018 to 113 in 2020. Voucher-based donations continued to grow throughout the pandemic. The following trends have been seen: (1) a steady rise in FV-based donation from the inception of the voucher program to present; (2) a steady shift in the percent involvement from standard altruistic donation to voucher-based donation; and (3) an annual decline in number of standard altruistic donors over the past 3 years. CONCLUSIONS: The following trends have been seen: (1) a steady rise in FV-based donation from the inception of the voucher program to present; (2) a steady shift in the percent involvement from standard altruistic donation to voucher-based donation; and (3) an annual decline in number of standard altruistic donors over the past 3 years. Though greater follow up is required, the short term trends suggest that voucher-based donation may be “cannibalizing” donation. Despite this, the overall number of non-directed donations, and the chains of transplants they spark, have expanded, delivering more high-quality kidneys and shortening the waiting time for kidney-paired exchange. Source of Funding: OneLegacy Foundation © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e597 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Nima Nassiri More articles by this author Torey Averick More articles by this author Jeffrey Veale More articles by this author Expand All Advertisement PDF DownloadLoading ...

The Kidney Donor Profile Index (KDPI) Correlates With Histopathologic Findings in Post-reperfusion Baseline Biopsies and Predicts Kidney Transplant Outcome.

BackgroundThe increasing organ shortage in kidney transplantation leads to the necessity to use kidneys previously considered unsuitable for transplantation. Numerous studies illustrate the need for a better decision guidance rather than only the classification into kidneys from standard or expanded criteria donors referred to as SCD/ECD-classification. The kidney donor profile index (KDPI) exhibits a score utilizing a much higher number of donor characteristics. Moreover, graft biopsies provide an opportunity to assess organ quality.MethodsIn a single center analysis 383 kidney transplantations (277 after deceased and 106 after living donation) performed between January 1st, 2006, and December 31st, 2016, retrospectively underwent SCD/ECD and KDPI scoring. Thereby, the quality of deceased donor kidneys was assessed by using the KDPI and the living donor kidneys by using the living KDPI, in the further analysis merged as (L)KDPI. Baseline biopsies taken 10 min after the onset of reperfusion were reviewed for chronic and acute lesions. Survival analyses were performed using Kaplan-Meier analysis and Cox proportional hazards analysis within a 5-year follow-up.ResultsThe (L)KDPI correlated with glomerulosclerosis (r = 0.30, p < 0.001), arteriosclerosis (r = 0.33, p < 0.001), interstitial fibrosis, and tubular atrophy (r = 0.28, p < 0.001) as well as the extent of acute tubular injury (r = 0.20, p < 0.001). The C-statistic of the (L)KDPI concerning 5-year death censored graft survival was 0.692. Around 48% of ECD-kidneys were classified as (L)KDPI<85%. In a multivariate Cox proportional hazard analysis including (preformed) panel reactive antibodies, cold ischemia time, (L)KDPI, and SCD/ECD-classification, the (L)KDPI was significantly associated with risk of graft loss (hazard ratio per 10% increase in (L)KDPI: 1.185, 95% confidence interval: 1.033–1.360, p = 0.025). Survival analysis revealed decreased death censored (p < 0.001) and non-death censored (p < 0.001) graft survival in kidneys with an increasing (L)KDPI divided into groups of <35, 35–85, and >85%, respectively.ConclusionWith a higher granularity compared to the SCD/ECD-classification the (L)KDPI is a promising tool to judge graft quality. The correlation with chronic and acute histological lesions in post-reperfusion kidney biopsies underlines the descriptive value of the (L)KDPI. However, its prognostic value is limited and underlines the urgent need for a more precise prognostic tool adopted to European kidney transplant conditions.

Extended criteria donors for robotic right hepatectomy: A propensity score matched analysis.

Robotic right donor hepatectomy (RDH) has been reported from experienced centers with reduced morbidity when compared to open RDH. However, outcomes in donors with large grafts/complex biliovascular anatomy are unknown. Out of 170 robotic RDH, 100 had one or more of the following: graft weight ≥800 g, type 2/3 portal vein, >1 bile duct or hepatic artery and inferior hepatic veins >5 mm requiring reconstruction (extended criteria donors [ExRDH]), while the remaining 70 had standard anatomy (SRDH). After propensity score matching, 66 ExRDH were compared with 66 SRDH. Additionally, all robotic RDH performed were analyzed in three temporal phases (60, 60, and 50). Peak AST and ALT were higher amongst donors and recipients in the ExRDH arm compared to SRDH. Other intraoperative parameters and postoperative complications were similar between the two groups. During the last phase, donors demonstrated reduction in duration of surgery, postoperative complications, and hospital stay while recipients showed decreased blood loss and hospital stay. Robotic right hepatectomy performed in donors with extended criteria have similar perioperative outcomes as standard donors. However, a significant learning curve needs to be traversed. Further studies are required before safely recommending robotic RDH for all donors.

Normothermic Ex Vivo Lung Perfusion (Novel) as an Assessment of Extended Criteria Donor Lungs: A Prospective Multi-Center Clinical Trial

<h3>Purpose</h3> <i>Ex vivo</i> lung perfusion (EVLP) allows re-evaluation of extended criteria/marginal donor lungs. This can increase the number of lung transplants. However, the long-term outcomes of transplanting EVLP-screened lungs in a multicenter setting are unknown. We proposed to evaluate the short- and long-term outcomes of EVLP performed at multiple centers. <h3>Methods</h3> This is a prospective, nonrandomized clinical trial. Seventeen lung transplant centers in the United States. Adult patients with end-stage pulmonary disease requiring lung transplant from May 2011 to December 2017 were eligible. Lung allografts initially deemed extended criteria/marginal (n=216) were placed on EVLP and re-evaluated prior to transplant. Patients received either standard donors (n=116) or lungs screened with EVLP (n=110). <h3>Results</h3> Half of the lung grafts (110/216, 50.9%) placed on EVLP were transplanted. The incidence of primary graft dysfunction 24 hours post-transplant was higher in the EVLP group (25.5% vs 10.3%, <i>p</i>=0.003), but was not significantly different 48 hours (EVLP: 15.5%, control: 9.5%, <i>p</i>=0.49) and 72 hours (13.6% vs 6.9%, p=0.34) post-transplant. Survival was not significantly different between the 2 groups 1 year (n=226, EVLP: 86%, control: 94%, <i>p</i>=0.06), 3 years (n=226, EVLP: 68%, control: 76%, <i>p</i>=0.16, Figure), or 5 years (n=159, EVLP: 59%, control: 65%, <i>p</i>=0.68) post-transplant. There were also no differences in pulmonary function, the incidence of chronic lung allograft dysfunction or quality of life measures post-transplant. <h3>Conclusion</h3> In this multicenter study, recipients of lungs that were re-evaluated on EVLP and deemed suitable for transplant had similar outcomes as a recipients of a standard lung transplants. EVLP offers the opportunity to screen donated lungs initially considered high risk and can safely increase the availability of transplantable lungs without compromising outcomes.

How to Assess EVLP Lungs in Standard Clinical Use?

<h3>Purpose</h3> Ex-vivo lung perfusion (EVLP) is currently considered a valuable option to preserve, reassess or recondition pulmonary grafts prior to lung transplantation (LTx). Both standard and extended criteria donors lungs have been studied in clinical trials. Usually, a 20% deterioration of physiological parameters during EVLP is considered detrimental for the graft and a reason to decline it. In this study, we report the relation between EVLP parameters and primary graft dysfunction (PGD) in a clinical LTx program. <h3>Methods</h3> In our center the TransMedics OCS^TM Lung device is used. The indication for EVLP was combined organ transplantation, graft re-assessment or prolonged preservation based on an institutional algorithm and available resources. Standard clinical EVLP parameters (mean pulmonary artery pressure (PAP), vascular resistance (VR) and driving airway pressure(DAP)) were compared between the first and final assessment. The % difference (Δ) between these time points was calculated. Graft outcome was expressed as presence of any PGD grade 3 (PGD 3) within 72 hours. Statistical analysis was performed using student T-test comparing patients with PGD 3 vs. without. Combined organ (liver-LTx) and LTx cases were analyzed separately as well as together. <h3>Results</h3> Data are presented in figure 1. In total, 12 lungs were included. Overall, all grafts met acceptance criteria during EVLP and were transplanted. Despite this, 58% of the patients developed PGD 3. No statistically significant difference in EVLP parameters was observed between patients who developed and did not develop PGD 3. <h3>Conclusion</h3> Overall, the clinical EVLP parameters were within the limits of acceptance and even improved over time. Despite this, we observed a high incidence of PGD 3. At this moment it is not clear how to assess EVLP lungs in clinical use and predict the outcome after LTx. The role of EVLP in post-reperfusion injury requires further research.

Consent to organ offers from public health service \u201cIncreased Risk\u201d donors decreases time to transplant and waitlist mortality

BackgroundThe Public Health Service Increased Risk designation identified organ donors at increased risk of transmitting hepatitis B, hepatitis C, and human immunodeficiency virus. Despite clear data demonstrating a low absolute risk of disease transmission from these donors, patients are hesitant to consent to receiving organs from these donors. We hypothesize that patients who consent to receiving offers from these donors have decreased time to transplant and decreased waitlist mortality.MethodsWe performed a single-center retrospective review of all-comers waitlisted for liver transplant from 2013 to 2019. The three competing risk events (transplant, death, and removal from transplant list) were analyzed. 1603 patients were included, of which 1244 (77.6%) consented to offers from increased risk donors.ResultsCompared to those who did not consent, those who did had 2.3 times the rate of transplant (SHR 2.29, 95% CI 1.88–2.79, p < 0.0001), with a median time to transplant of 11 months versus 14 months (p < 0.0001), as well as a 44% decrease in the rate of death on the waitlist (SHR 0.56, 95% CI 0.42–0.74, p < 0.0001). All findings remained significant after controlling for the recipient age, race, gender, blood type, and MELD. Of those who did not consent, 63/359 (17.5%) received a transplant, all of which were from standard criteria donors, and of those who did consent, 615/1244 (49.4%) received a transplant, of which 183/615 (29.8%) were from increased risk donors.ConclusionsThe findings of decreased rates of transplantation and increased risk of death on the waiting list by patients who were unwilling to accept risks of viral transmission of 1/300–1/1000 in the worst case scenarios suggests that this consent process may be harmful especially when involving “trigger” words such as HIV. The rigor of the consent process for the use of these organs was recently changed but a broader discussion about informed consent in similar situations is important.

Iliac Crest Bone Graft Harvesting: Modified Technique for Reduction of Complications.

bone grafting; complications; iliac crest; pain.

Utilizing Ternary Strategy to Reduce the Influence of Polymer Batch‐to‐Batch Variation in Organic Solar Cells

Despite producing the best device performances, several major issues such as the batch‐to‐batch variation and low molecular weight (M w) optimization greatly hinder the progress for the organic solar cells. Herein, tetraphenylethylene (TPE) as a third component has been added in a series of PM6:Y6 blends with different donor molecular weights (42–129 kDa). Ultimately, ternary devices fabricated by the PM6 with about half the M w (66 and 74 kDa) of the standard donor (129 kDa) produce an excellent power conversion efficiency (PCE) of 15.09% and 15.15% owing to enhanced morphology and charge mobility, as compared to a standard binary PCE of 15.65%, respectively, indicating the possibility of attaining high‐performance attributes from low M w polymers. Furthermore, the TPE inclusion in the 129 kDa binary blend leads to a remarkable PCE of 16.48%. This strategy, therefore, illustrates a relatively simple approach to overcome the batch‐to‐batch variation problem associated with polymers for organic solar cells.

Development of sensitive and specific loop-mediated isothermal amplification combined with lateral flow device for the rapid detection of hepatitis B virus infection.

HBV standard plasma panels and donor plasma specimens were used to evaluate the assay. HBV DNA was extracted by using QiAamp DNA Blood Mini Kit. Amplification was carried out at constant temperature 63°C for 60min. LAMP end products were analyzed by using ESE LAMP tube scanner, gel electrophoresis, UV-lamp, and lateral flow device. HBV-LAMP-LFD assay revealed sensitivity of 92% (138/150) of HBV positive plasma specimens. Specificity of HBV-LAMP-LFD was calculated 100%. Our study concludes that HBV-LAMP-LFD is rapid, easy to use, sensitive, and specific point-of-care diagnostic assay for the detection of hepatitis B virus in blood samples. This assay can be used in resource-limited settings as well as in HBV endemic areas.

Early experiments with hypothermic oxygenated machine perfusion of kidney grafts from extended criteria donors

Objective: to evaluate the safety and efficacy of hypothermic oxygenated machine perfusion (HOPE) for kidney grafts obtained from expanded criteria donors (ECD).Materials and methods. From June 2018 to June 2021, 200 surgeries involving kidney transplants from deceased donors were performed at Botkin City Clinical Hospital. Of these, 123 were men (61.5%) and 77 were women (38.5%). The mean age was 47.62 ± 11.69 (20–73) years. In 102 cases, kidney grafts were procured from ECD. In 92 recipients (90.2%) of kidney transplants from an expanded criteria donor, static cold storage done according to the standard technique was used to preserve the organ; these patients constituted observation group 1. In 10 recipients (9.8%), hypothermic oxygenated perfusion was used in addition to static cold preservation; these patients formed observation group 2.Results. No 30-day mortality was recorded in both observation groups. The mean static cold storage time in group 1 patients was 612.33 ± 178.88 (133–1180) minutes. Overall incidence of delayed graft function was 26.5% (53/200). Incidence of delayed graft function was 19.3% (19/98) for organs from standard donors using static cold storage and 35.8% (33/92) for ECD organs. Twenty-five patients (12.5%) had postoperative complications. Postoperative complications with delayed graft function were diagnosed in 12 patients, which was 22.6% (12/53), with immediate function in 13 patients, which was 8.8% (13/147). Mean cold storage time in group 2 patients was 319.11 ± 110.24 (311–525) minutes. Mean HOPE time was 202.34 ± 21.48 (150–210) minutes. Delayed graft function was recorded in 1 group 2 patient (10%). No complications, including perfusion-related one, were recorded in this group.Conclusion. The unique technique used at Botkin City Clinical Hospital for HOPE in kidney transplant is safe. It provides a low risk of delayed graft function for ECD kidneys.

Deep Learning Algorithms for the Prediction of Posttransplant Renal Function in Deceased-Donor Kidney Recipients: A Preliminary Study Based on Pretransplant Biopsy.

BackgroundPosttransplant renal function is critically important for kidney transplant recipients. Accurate prediction of graft function would greatly help in deciding acceptance or discard of allocated kidneys.Methods: Whole-slide images (WSIs) of H&E-stained donor kidney biopsies at × 200 magnification between January 2015 and December 2019 were collected. The clinical characteristics of each donor and corresponding recipient were retrieved. Graft function was indexed with a stable estimated glomerular filtration rate (eGFR) and reduced graft function (RGF). We used convolutional neural network (CNN)-based models, such as EfficientNet-B5, Inception-V3, and VGG19 for the prediction of these two outcomes.ResultsIn total, 219 recipients with H&E-stained slides of the donor kidneys were included for analysis [biopsies from standard criteria donor (SCD)/expanded criteria donor (ECD) was 191/28]. The results showed distinct improvements in the prediction performance of the deep learning algorithm plus the clinical characteristics model. The EfficientNet-B5 plus clinical data model showed the lowest mean absolute error (MAE) and root mean square error (RMSE). Compared with the clinical data model, the area under the receiver operating characteristic (ROC) curve (AUC) of the clinical data plus image model for eGFR classification increased from 0.69 to 0.83. In addition, the predictive performance for RGF increased from 0.66 to 0.80. Gradient-weighted class activation mappings (Grad-CAMs) showed that the models localized the areas of the tubules and interstitium near the glomeruli, which were discriminative features for RGF.ConclusionOur results preliminarily show that deep learning for formalin-fixed paraffin-embedded H&E-stained WSIs improves graft function prediction accuracy for deceased-donor kidney transplant recipients.

A Review of Current and Emerging Trends in Donor Graft-Quality Assessment Techniques.

The number of patients placed on kidney transplant waiting lists is rapidly increasing, resulting in a growing gap between organ demand and the availability of kidneys for transplantation. This organ shortage has forced medical professionals to utilize marginal kidneys from expanded criteria donors (ECD) to broaden the donor pool and shorten wait times for patients with end-stage renal disease. However, recipients of ECD kidney grafts tend to have worse outcomes compared to those receiving organs from standard criteria donors (SCD), specifically increased risks of delayed graft function (DGF) and primary nonfunction incidence. Thus, representative methods for graft-quality assessment are strongly needed, especially for ECDs. Currently, graft-quality evaluation is limited to interpreting the donor’s recent laboratory tests, clinical risk scores, the visual evaluation of the organ, and, in some cases, a biopsy and perfusion parameters. The last few years have seen the emergence of many new technologies designed to examine organ function, including new imaging techniques, transcriptomics, genomics, proteomics, metabolomics, lipidomics, and new solutions in organ perfusion, which has enabled a deeper understanding of the complex mechanisms associated with ischemia-reperfusion injury (IRI), inflammatory process, and graft rejection. This review summarizes and assesses the strengths and weaknesses of current conventional diagnostic methods and a wide range of new potential strategies (from the last five years) with respect to donor graft-quality assessment, the identification of IRI, perfusion control, and the prediction of DGF.

Nanolayer Laser Absorber for Femtoliter Chemistry in Polymer Reactors.

Laser-induced forward transfer (LIFT) has the potential to be an alternative approach to atomic force microscopy based scanning probe lithography techniques, which have limitations in high-speed and large-scale patterning. However, traditional donor slides limit the resolution and chemical flexibility of LIFT. Here, a hematite nanolayer absorber for donor slides to achieve high-resolution transfers down to sub-femtoliters is proposed. Being wettable by both aqueous and organic solvents, this new donor significantly increases the chemical scope for the LIFT process. For parallel amino acid coupling reactions, the patterning resolution can now be increased more than five times (>111000 spotscm- 2 for hematite donor vs 20000 spotscm- 2 for standard polyimide donor) with even faster scanning (2vs 6ms per spot). Due to the increased chemical flexibility, other types of reactions inside ultrasmall polymer reactors: copper (I) catalyzed click chemistry and laser-driven oxidation of a tetrahydroisoquinoline derivative, suggesting the potential of LIFT for both deposition of chemicals, and laser-driven photochemical synthesis in femtoliters within milliseconds can be explored. Since the hematite shows no damage after typical laser transfer, donors can be regenerated by heat treatment. These findings will transform the LIFT process into an automatable, precise, and highly efficient technology for high-throughput femtoliter chemistry.

Donor Milk Banking - Safety, Efficacy, New Methodologies.

Donor milk (DM) is of increasing interest as primary nutritional source for preterm infants. Safe access requires special infrastructure, trained staff, sophisticated algorithms, and standard operating procedures as well as quality control measures. DM has limitations like low protein content and unpredictable composition of the other macronutrients, despite fortification frequently not meeting recommendations - both of them compromising growth. The first paragraph is devoted to COVID-19 and how it impacts processes of DM banking. The following paragraphs review aspects of "pasteurization," "safety audits/donor screening," and "DM nutrient variability." In summary, (i) Holder pasteurization still is the most suitable procedure for milk banks, but high-pressure pasteurization or ultraviolet C irradiation conserve the unique properties of DM better and deserve more research to make it suitable for clinical routine. (ii) In regard to safety/screening, guidelines are valuable for safe DM bank operation, but they differ between legislations. There is a surprisingly high rate of non-disclosed donor smoking (0.3%, p > 0.05) and of adulteration of delivered DM (up to 2%, p < 0.05) not detected by standard donor screening procedures. Frequencies differ between remunerated and non-remunerated programs. (iii) Neonatal caregivers should be aware of unpredictable composition of DM. They should be trained on how these can be overcome to avoid negative impact on growth and long-term outcomes like (a) measuring and disclosing nutrient contents of delivered DM batches to customers, (b) implementing certain types of donor pooling to reduce the risk of macronutrient depleted DM, (c) additional supplementation using 0.3-0.5 g protein/100 mL seems to be reasonable, (d) adjusted fortification may help to improve growth, but is not efficient in all preterm infants, (e) target fortification seems to improve growth (and probably also neurodevelopmental index) compared to standard fortification, (f) more research and clinical studies are needed.

Development of a cost-effective ex vivo lung perfusion system for lung transplantation in India.

Standard donor lung preservation with cold flush and storage allows up to six hours between retrieval of lungs from the donor and transplantation in the recipient. Ex vivo lung perfusion (EVLP) systems mimic physiological ventilation and perfusion in the donor lungs with potential for prolonged lung preservation and donor lung reconditioning. In this study, it was aimed to perform EVLP on discarded donor lungs using a locally developed EVLP system. Equipment that are routinely used for cardiac surgeries were collected and a functional EVLP system was assembled. This system was used on five pairs of lungs retrieved from brain-dead organ donors. The lungs were ventilated and pulmonary circulation was continuously perfused with a solution containing oxygen and nutrients for four hours. The system was tested without red blood cells (RBCs) added to the solution (acellular group; n=3; A1, A2 and A3) and also with RBCs added to the solution (cellular group; n=2; C1 and C2). The EVLP system was successfully used in four (A1, A2, A3 and C2) of the five lung pairs. Mechanical and gas exchange functions of the lungs were preserved in these lung pairs. One lung pair (C1) worsened and developed pulmonary oedema. Histopathological examination of all five lung pairs was satisfactory at the end of the procedure. Major challenges faced were leakage of solution from the system and obstruction to drainage of RBCs containing solution from the lungs. The results of the present study suggest that, it is possible to maintain the lungs retrieved for transplantation in a physiological condition using a locally prepared EVLP system and a solution without RBCs.

Introducing robot‐assisted laparoscopic donor nephrectomy after experience in retroperitoneal endoscopic approach: a matched propensity score analysis

To assess the safety and efficacy of introducing robotic-assisted laparoscopic donor nephrectomy (RALDN) to the standard retroperitoneal endoscopic donor nephrectomy (REDN). Data were collected prospectively from 124 consecutive living kidney donors (93 for REDN subgroup and 31 for RALDN subgroup) from February 2018 to December 2020. Donor baseline demographics, perioperative outcomes and recipient outcomes were recorded, and these parameters were compared between the two subgroups before and after propensity-score matching. Mean age was 51.1 ± 9.1 years; 42.7% were males; mean body mass index was 22.7 ± 2.4; and there were 109 (88%) left kidneys. The following data of REDN and RALDN was, respectively, recorded: operative time (213 ± 43 versus 216 ± 39 min, p=0.721), warm ischemic time (4.7 ± 1.2 versus 4.9 ± 1.4 min, p=0.399), postoperative complications (5.4% versus 6.5%, p=1), haemoglobin (g/L) drop (9.4 ± 7.2 versus 9.7 ± 6.6, p=0.836), blood creatinine at 6 month (1.15 ± 0.23 versus 1.13 ± 0.24 mg/dL, p=0.734) and at 1 year (1.09 ± 0.22 versus 1.17 ± 0.28 mg/dL, p=0.591). In post-propensity score matched analyses, there was no significant differences between the two groups including intraoperative and postoperative complications. RALDN could be safely introduced into a living donor program experienced in laparoscopic donor nephrectomy. The outcomes of our study comparing these minimally invasive techniques are mostly similar in terms of intraoperative and postoperative outcomes for kidney donors.

Donor memory-like NK cells persist and induce remissions in pediatric patients with relapsed AML after transplant

AbstractPediatric and young adult (YA) patients with acute myeloid leukemia (AML) who relapse after allogeneic hematopoietic cell transplantation (HCT) have an extremely poor prognosis. Standard salvage chemotherapy and donor lymphocyte infusions (DLIs) have little curative potential. Previous studies showed that natural killer (NK) cells can be stimulated ex vivo with interleukin-12 (IL-12), -15, and -18 to generate memory-like (ML) NK cells with enhanced antileukemia responses. We treated 9 pediatric/YA patients with post-HCT relapsed AML with donor ML NK cells in a phase 1 trial. Patients received fludarabine, cytarabine, and filgrastim followed 2 weeks later by infusion of donor lymphocytes and ML NK cells from the original HCT donor. ML NK cells were successfully generated from haploidentical and matched-related and -unrelated donors. After infusion, donor-derived ML NK cells expanded and maintained an ML multidimensional mass cytometry phenotype for >3 months. Furthermore, ML NK cells exhibited persistent functional responses as evidenced by leukemia-triggered interferon-γ production. After DLI and ML NK cell adoptive transfer, 4 of 8 evaluable patients achieved complete remission at day 28. Two patients maintained a durable remission for >3 months, with 1 patient in remission for >2 years. No significant toxicity was experienced. This study demonstrates that, in a compatible post-HCT immune environment, donor ML NK cells robustly expand and persist with potent antileukemic activity in the absence of exogenous cytokines. ML NK cells in combination with DLI present a novel immunotherapy platform for AML that has relapsed after allogeneic HCT. This trial was registered at https://clinicaltrials.gov as #NCT03068819.