Innovative technologies require the use of materials that meet increasingly high requirements; one such requirement is the purity of metals. In the case of copper, this translates into a parameter related to electrical conductivity. Traditional metal refining technologies have some limitations that can be eliminated through the use of modern melting aggregates. Such solutions include vacuum induction furnaces, comprising an induction furnace with a cold crucible. As part of this work, the possibilities of refining copper and lead alloys were investigated. In addition, the research was carried out with the use of two induction vacuum aggregates, allowing us to compare their effectiveness. The tests were carried out in a pressure range of 10–1000 Pa and at temperatures of 1273–1473 K. The results obtained made it possible to determine the mass transport coefficient of lead from an alloy with copper, and to determine the share of resistance in individual stages of the process. For experiments conducted inside an induction crucible furnace, lowering the working pressure inside the furnace chamber from 1000 to 10 Pa while increasing the temperature from 1323 to 1473 K was accompanied by a drop in the lead concentration inside the alloy of 69 to 96%, compared to its initial mass. For experiments conducted inside a cold crucible furnace, approximate values of lead removal appeared for lower temperatures (1273 to 1323 K), confirming that the analyzed process happens faster in this aggregate. In recent years, demand for metallic materials with special properties has increased. It has forced the development of technologies that allow for the production of metals and alloys of high purity. At the same time, the significance of vacuum smelting technology has also grown. Vacuum metal refinement technologies are based on the phenomena of the evaporation of metal bath components characterized by high vapor pressure, and the separation of gasses contained within the metal bath. The process of evaporation is complex, due to its heterogenic character. The temperature and pressure within the molten aggregate, its fluid dynamics, and the alloy chemical structure can strongly impact the rate of the analyzed process. Determining which of these properties has the most significant impact on the kinetics of the evaporation process and determining the character of the control of this process require multiple experiments to be conducted.Vacuum induction melting (VIM) is a commonly used melting technique which involves melting an alloy in a vacuum or in an inert atmosphere by electromagnetic induction using coils. The main source of power during the process is the alternative magnetic field formed by an alternate current passing through an induction coil. Compared to other techniques, the use of VIM technology for metals alloys in refractory crucibles is much less energy intensive ,2,3. In In the work presented here, we show the examination results of the rate with which lead was removed from copper alloys molten within a vacuum induction crucible furnace (ICF) and a cold crucible furnace (CCF). This specific type of examination was conducted due to the fact that the contamination examined strongly lowers the electric conductivity of copper. The devices used were chosen because induction smelting causes a significant increase in the surface of the metal bath, which can intensify the analyzed process of refining copper from lead.The first literature references related to the research of copper or copper alloy vacuum refining were published in the 1960s and in the first half of the 1970s ,12,13,14Further papers published by Ozberk and Guthrie ,18 contaFrom these works, only  addresseExaminations were conducted on copper alloys, the compositions of which are shown in The research programme was realized using two vacuum induction furnaces manufactured by SECO-WARWICK : namely, a vacuum induction crucible furnace ICF (with a ceramic crucible) and an induction furnace with a cold crucible CCF. The baseline technical and construction parameters of both devices are shown in A sample of the examined alloy with a mass of 3300 g was placed inside the crucible. Depending on the type of the furnace, it was either an Isopress 1126-MG951 crucible produced by Capital Refractories made out of MgO, or a cold crucible cooled with water made from copper. The inside diameter of both crucibles was equal to 90 mm. Each of the experiments described in this work was conducted following the same approach. In the first stage, after inserting the sample into the furnace, the pressure was lowered inside the chamber and then heating was turned on and the insert was melted. After achieving the desired temperature of liquid metal, it was kept inside the furnace for 600 s. The temperature was measured using a thermoelectric sensor, type PtRh30-PtRh6, and an optical pyrometer. During the process, samples of liquid metal were extracted at the defined time intervals. After the experiment was finished, the chemical composition of the metal was examined. For this purpose, atomic absorption spectrometry was used, using an ASA solar device.In the ICF aggregate, the process was analyzed for the temperature range of 1323–1473 K; for the CCF aggregate, the temperature range was 1273–1323 K. The furnace’s working power was in the range of 30–40 kW for the ICF, and it was equal to 170 kW for the CCF. The working pressure for both aggregates varied within the range of 10 to 1000 Pa.kPb) in the analyzed process can be described using Equation (1):lβ—the mass transfer coefficient of Pb in liquid phase, m·s−1,gβ—the mass transfer coefficient of Pb in gas phase, m·s−1,ek—the evaporation rate coefficient, m·s−1.The overall coefficient of lead mass transport :F—the evaporation surface, m2,V—the metal volume, m3,t − t0)—the duration of the process, s., m,DPb—the lead diffusion coefficient in liquid copper, m2 s−1,In the case of metal that is melted and mixed inside an induction furnace, to calculate the coefficient value for the transport of lead inside the liquid phase equation  is recomkePb can be described by Equation (5):α—the evaporation constant,p0Pb—the equilibrium vapor pressure of lead over the liquid alloy, Pa,γPb—the activity factor of lead in liquid copper,MCu, MPb—the molar masses, respectively, of copper and lead, g mol−1,ρCu—the density of copper, g m−3.Analyzing the evaporation rate of the load from the liquid copper surface, it was assumed that the maximum value of the rate constant Rl) and the resistance related to the reaction happening on the liquid alloy surface (Re) in the overall evaporation process resistance, Equations (6) and (7) were used:To estimate the resistance values for the process of mass transport in the liquid phase (CkPb), the percentage mass loss of lead (m), and the calculated values of the evaporation stream of lead (N). A graphical interpretation of these results is shown in Analyzing the results shown in −4 g/(cm2 s) and 3.54 × 10−4 g/(cm2 s), and for the cold crucible the samples were between 2.23 × 10−4 g/(cm2 s) and 1.67 × 10−4 g/(cm2 s). This was caused by the significant difference in the size of the molten metals in both aggregates.Additionally, the study results show an increase in the effectiveness of the lead removal process as the pressure was lowered within the smelting aggregate. At the same time, there was a strong difference in the density of the stream of removed lead. The approximated values for this parameter for samples for the crucible furnace were between 6.14 × 10To measure the stages that determine the rate of the examined evaporation process, kinetic analysis was conducted, based on its experimental results.Lead transport from inside liquid copper to the surface;The phenomenon of evaporation from the surface of liquid copper;The transport of lead vapor into the gas phase.From a kinetic point of view, the process of lead evaporation from liquid copper can be divided into three stages:kPb, knowledge of both the size of the bath and its surface area are required. In the experiments analyzed, in the temperature range of 1273–1473 K, the estimated value of the total liquid metal volume was in the 1.21 to 1.26 × 10−4 m3 range. The density values of liquid copper in the analyzed temperature range were calculated based on the relations contained within the work [From Equation (3) it can be concluded that, to calculate the value of the coefficient the work . A hardethe work ,29. ThisCreating photos of molten metal using a fast camera;Establishing the geometry of the created meniscus based on these photos;Charting a function graph that describes the meniscus during different parameters of the process;Estimating the surface area using Wolfram Mathematica and MicroStation software.To estimate the real surface size of the liquid metal, the method described in the paper ,31 was u2 for samples inside a crucible furnace, and between 227 and 230 cm2 for experiments inside a cold crucible furnace. For comparison, the inside surface area of the crucible used measured at 63.6 cm2.Estimations of metal surface area based on the described method are variable within the range of 113 to 131 cmPb are shown in kPb coefficient as a function of the working pressure of an induction crucible furnace.Calculated from Equation (3), the values of the overall lead transport coefficient kkPb were within the range of 1.66 × 10−5 to 5.80 × 10−5 ms−1. These results concern both the experiments conducted inside an inductive crucible furnace and those conducted inside a cold crucible furnace. Reducing the working pressure of the melting aggregates and the temperature caused the increase in the value of this coefficient, which means a higher rate of lead evaporation. The kPb coefficient values achieved during the study are in line with the ones in the table.From the data shown in βl is directly proportional to the value of the surface rate of the liquid metal. Most authors researching the evaporation kinetics of inductively mixed metal bath ingredients assume, as Machlin did, that this rate is not related to the electric parameters of the furnace. For induction furnaces with volumes up to one ton, it was assumed that the mν is constant and measures at 0.1 ms−1. However, later works determined that this rate is related to parameters such as furnace power, current frequency, crucible geometry, or the position of the crucible inside the furnace compared to the inductor [βl coefficient, rate values were taken from [m, the rate values were between 0.114 and 0.135 ms−1. In βl coefficient values are shown, and were calculated using Equation (4). They fell within the range of 1.40–1.78 × 10−4 ms−1. The lead diffusion coefficients from liquid copper were estimated using [Machlin assumed, based on the analogy of penetrative mass insertion models, that elements of liquid metal move according to a tangent: liquid metal–gas or liquid metal–crucible, whereby the gradient of the rate normal to the surface is close to zero. From this correlation, it can be concluded that the value of the inductor ,33. In tken from , which wed using .p0Pb presented in this work were calculated based on thermodynamics [Pbγ. Based on these estimates, for the temperature range of 1273–1473 K, the values of p0Pb ranged between 0.91 × 102 and 1.91 × 103 Pa, and the activity coefficient values ranged from 4.52 to 7.25. The kePb coefficient values calculated from Equation (5) are shown in The values of lead vapor pressure dynamics ; data frdynamics  were useRl and Re in the overall resistance of the evaporation process, depending on the working pressure inside the melting aggregate.Re resistance on the overall process resistance is between 9 and 28%Based on the data shown in Re and resistance of mass transport inside the liquid phase Rl in the overall resistance of the evaporation process of lead were within the range of 21 to 61%. The summarized values of resistance Rl + Re exceeded 50% for tests conducted at a pressure of 10 Pa.For tests conducted inside the crucible furnace, the overall resistance contribution of transport resistance related to the reaction happening on the surface of liquid copper Rl in the overall resistance of the lead evaporation process did not exceed 34%. This means that, in this case, the lead evaporation rate is determined mainly by mass transport in the gaseous phase.For tests conducted inside a furnace with a cold crucible, the summarized mass transport resistance in the liquid phase The process of refining copper from lead conducted in vacuum induction furnaces happens faster inside an aggregate with a cold crucible, as compared to a typical crucible furnace. This is caused by a significant difference in the size of the metal bath’s surface area. The calculated surface areas of the meniscus formed during smelting alloys using a furnace with a cold crucible are significantly larger compared to those where the inserts were melted in induction crucible furnaces.Pbk increased from 3.50 × 10−5 to 5.27 × 10−5 ms−1.For the experiments conducted inside an induction crucible furnace, lowering the working pressure inside the furnace chamber from 1000 to 10 Pa while increasing the temperature from 1323 to 1473 K was accompanied by a drop in the lead concentration inside the alloy of between 69 and 96%, compared to its initial mass. At the same time, the value of the overall mass transport coefficient For the experiments conducted inside a cold crucible furnace, approximate values of lead removal appeared for lower temperatures (1273 to 1323 K), confirming that the analyzed process happens faster in this aggregate.lR) and the reaction on the surface of the liquid metal (eR) comprises nearly 60% of the overall process resistance.When the pressure is lowered, the rate-determining step of the analyzed process changes. When the pressure is equal to 10 Pa, the value of resistance related to mass transport in the liquid phase (For the experiments conducted inside a cold crucible surface, in the overall temperatures and pressures used, the rate of the process is limited by mass transport in the gas phase.The presented results allow us to formulate the following conclusions:


Limbo-keratoplasty enables visual improvement and limbal stem cell transplantation at the same. During follow-up, most grafts show vascularization of the limbus. However, it is unclear whether vascularization is harmful due to immunologic effects or helpful to nourish the limbal stem cells and is therefore necessary for a clear graft. The aim of our study is to analyze the influence of graft vascularization on graft survival following homologous limbo-keratoplasty.In this retrospective study, we assessed all consecutive limbo-keratoplasties performed in our hospital. All eyes with suitable photo-documentation were included and divided into two groups . We categorized the grade of vascularization  and analyzed clear graft survival, recurrence of the underlying disease and the endothelial cell density (ECD) with regard to the reason for the graft. Event rates were estimated with the Kaplan–Meier method.A total of 79 eyes with limbal stem cell deficiency and 15 with corneal dystrophies were analyzed. A high degree of graft vascularization had a tendency for better graft survival in limbal stem cell deficiency, whereas in corneal dystrophies, grafts with no vascularization had preferable outcomes. Recurrence-free graft survival was only seen in grade 1 and 3 vascularization in corneal dystrophies.Vascularization of the limbus seems to have an impact on the long-term outcome of limbo-keratoplasty. The effect seems to be favorable in limbal stem cell deficiency and on recurrence rates in corneal dystrophies. However, the latter might be overshadowed by an unfavorable immunologic effect in corneal dystrophies where the baseline immunologic risk profile is commonly more favorable than in limbal stem cell deficiency. Allogenic limbo-keratoplasty was first introduced by Sundmacher et al. . The aimIt has been shown that fewer transplants develop a recurrence of the dystrophy if limbal tissue is also transplanted . The sim2. Statistical analysis was done with R (R-project.org). Due to the retrospective exploratory character of our study along with the small sample size we decided to perform a descriptive presentation of the data rather than testing for statistical significance. Ethics committee approval was obtained at the ethics committee of the Albert-Ludwigs University Freiburg (Nr. 28/19).The data of all consecutive limbo-keratoplasties performed at our institution from 2003 to 2020 were included in this retrospective study. All patients had topical and systemic immunosuppression if possible. Topical therapy usually consisted of dexamethasone eye drops. Systemic immunosuppression was usually a combination of cyclosporine and mycophenolate mofetil. In case of repeat transplantations in one patient, only the first one was included for analysis. We identified all cases with suitable photo-documentation of the graft that enabled to analyze the vascularization of the grafted limbal tissue. Only vascularization of the limbus was graded, not vascularization as indication to graft failure. The grade of vascularization was performed by two independent investigators according to example pictures of different grades of vascularization Fig. . Grafts A total of 206 eyes were assessed for this study. In total, 112 of those eyes had suitable postoperative photo-documentation. Ninety-four eyes did not have suitable photo-documentation, 79 with limbal stem cell deficiency and 15 with corneal dystrophies. All patients were treated with topical steroids. In the group of limbal stem cell deficiency, 89% were treated with systemic mycophenolate mofetil and 81% were treated with systemic ciclosporin. This results in a combination of both in 64% of the patients. In the group of corneal dystrophies, 82% were treated with systemic mycophenolate mofetil and 54% were treated with systemic ciclosporin. This results in a combination of both in 36% of the patients. Median age in the group with limbal stem cell deficiency was 51.9 years. The median age in the group with corneal dystrophies was 52 years. 43% of the patients were female. Median follow-up was 2.6 years in the group with limbal stem cell deficiency and 5.2 years in the group with corneal dystrophies. There were more eyes with a low grade of vascularization in the group with corneal dystrophies. There was no graft without vascularization in the group with limbal stem cell deficiencies  in the corneal dystrophy group and 0.009  in the limbal stem cell deficiency group. Postoperative visual acuity was 0.32  in the corneal dystrophy group and 0.01  in the limbal stem cell deficiency group.p = 0.1). Corneal dystrophies showed longer clear graft survival in general with 90% clear graft survival after 5 years. All grafts with grade 0 remained clear during complete follow-up. The graft survival decreased in the groups with higher vascularization with the lowest in grade 3 and 3b with no remaining clear grafts after 11 years  that maintained more than 1000 endothelial cells per mm-up Fig. .Fig. 4KaCompared to previous studies, our results show a comparable or slightly better overall graft survival , 11 for Regarding the graft survival in relation to the grade of vascularization of the grafted limbus, a difference between both indications was visible. In limbal stem cell failure, grafts with a higher degree of limbal vascularization tended to have a longer graft survival. In the group of corneal dystrophies, there was a tendency to a higher graft survival with no or low vascularization of the transplanted limbal tissue. The latter is in concordance with the literature where a higher degree of vascularization of the graft is usually associated with higher risk of graft failure due to immune reactions , 13. It The high recurrence rate of corneal dystrophy in our population seems rather early in comparison to the literature, where usually recurrence is seen later –16. Graf2 in the study of Khattak and Nakhli [2 after 10 years in the study of Lass et al. [Regarding the corneal endothelium, there is a steady loss of endothelial cells in all grafts following limbo-keratoplasty independent of the vascularization grades in the corneal dystrophy group and in all but one in the group with limbal stem cell deficiency. Reports in the literature show that such a chronic loss of endothelial cells has to be expected and is well within usual range, with an median endothelial cell count of 1035 cells/mmd Nakhli  and 550 s et al. . In our Limitations of our study lie in its retrospective design, the limited number of patients and the possible bias in selection, since we could only include cases that had adequate photo-documentation. The selection bias might have excluded cases with very early graft failure. Also, the follow-up is relatively short. This is due to the retrospective nature of the study. Patients that live far away from our clinic might have their regular examinations close to home. We evaluated the overall clear graft survival, because a clear distinction between an endothelial graft failure and a failure of the transplanted limbus is not always possible in retrospect. Also, a certain distinction between status post-graft rejection and chronic endothelial cell loss was not possible due to the retrospective design.In conclusion, the vascularization of the transplanted limbal tissue seems to have an impact on the long-term outcome of the limbo-keratoplasty grafts. The effect seems to be favorable in limbal stem cell deficiency and on the recurrence in corneal dystrophies. However, the latter might be overshadowed by an unfavorable immunologic effect in corneal dystrophies where the baseline immunologic risk profile is commonly more favorable than in limbal stem cell deficiency. Future prospective studies are needed to verify these results and explore possible therapeutic applications.


CXCL1 belongs to a member of the ELR + CXC chemokine subgroups that also known as GRO-alpha. It has been recognized that several types of human cancers constitutively express CXCL1, which may serve as a crucial mediator involved in cancer development and metastasis via an autocrine and/or paracrine fashion. However, the expression pattern and clinical significance of CXCL1 in human uterine cervix cancer (UCC), as well as its roles and mechanisms in UCC tumor biology remains entirely unclear.The expression and clinical significance of CXCL1 in UCC tissues was explored using immunohistochemistry and bioinformatics analyses. The expression and effects of CXCL1 in HeLa UCC cells were assessed using ELISA, CCK-8 and transwell assays. Western blotting experiments were performed to evaluate the potential mechanism of CXCL1 on malignant behaviors of HeLa UCC cells.The current study demonstrated that CXCL1 was expressed in HeLa UCC cells, PHM1-41 human immortalized cervical stromal cells, as well as cervical tissues, with UCC tissues having an evidently high level of CXCL1. This high level of CXCL1 in cancer tissues was notably related to poor clinical stages and worse survival probability, rather than tumor infiltration and patient age. In addition, CXCL1 expression was extremely correlated with CCL20, CXCL8 and CXCL3 cancer-associated chemokines expression. In vitro, the growth and migration abilities of HeLa cells were significantly enhanced in the presence of exogenous CXCL1. Gain-function assay revealed that CXCL1 overexpression significantly promoted growth and migration response in HeLa cells in both autocrine and paracrine manners. Finally, we found that CXCL1 overexpression in HeLa cells influenced the expression of ERK signal-related genes, and HeLa cell malignant behaviors derived from CXCL1 overexpression were further interrupted in the presence of the ERK1/2 blocker.Our findings demonstrate the potential roles of CXCL1 as a promoter and a novel understanding of the functional relationship between CXCL1 and the ERK signaling pathway in UCC.The online version contains supplementary material available at 10.1186/s12885-022-09749-0. Uterine cervix cancer (UCC), a common malignancy, represents the fourth most frequently occurring cancer and the fourth leading cause of cancer-related death in females worldwide . It is eChemokines or chemoattractant cytokines are categorized into four subfamilies, namely CXC, CC, C, and CX3C, based on the number and spacing of the conserved cysteine residues near the N-terminus. The essential roles of chemokines in biology are mediated through their combinatorial interaction with the G protein-coupled chemokine receptors . Early sCXCL1, also known as GRO-alpha, is a member of the ELR + CXC chemokine subgroups characterized by the presence of a conserved glutamic acid-leucine-arginine (ELR) motif . CXCL1-rIn this study, we aimed to determine the expression pattern of CXCL1 protein in a commercial uterine cervical tissue microarray, as well as to evaluate its relationship with clinical characteristics. We also explored the potential exogenous autocrine and paracrine mechanisms involved in CXCL1-mediated UCC tumor biology, possibly via mitogen activated protein kinases (MAPK)/extracellular signal-regulated kinase (ERK)-dependent signal pathway.A commercial uterine cervical tissue microarray  with 40 cancer tissues and 40 non-cancerous tissues was used to evaluate CXCL1 protein expression. Cancer samples ranged in age from 22 to72 years while non-cancer samples ranged in age from 32 to 68 years. The cancer tissue samples were classified into 3 clinical stages and 3 pathological grades. The sample characteristics are shown in Table http://ualcan.path.uab.edu/) was employed to explored the association of CXCL1 or CXCR2 mRNA expression with patient survival. To further elucidate the positive genes that associated with CXCL1 signal in UCC, we further explored the correlation between CXCL1 and other genes expression using this database.UALCAN database , according to the manufacturer’s instructions. Finally, the transfection efficiency of CXCL1 in HeLa and PHM1-41 cells was assessed using ELISA.HeLa human cervical cancer cells and PHM1-41 human immortalized cervical stromal cells were obtained from American type culture collection . HeLa and PHM1-41 cells were respectively cultured in RPMI‑1640 and Dulbecco’s Modified Eagle’s Medium  supplemented with 10% fetal bovine serum  containing antibiotics (100 units/ml penicillin and 100 mg/ml streptomycin) in a 37 °C humidified incubator with 5% COThe culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit  following the manufacturer’s instruction. Briefly, after culture for 24 h, cell culture media were collected. 100 μL of culture supernatant and different concentrations of recombinant CXCL1 standard protein were added to each well of the plate. The plate was then incubated at 37 °C for 90 min. Following triple rinsing with washing buffer, the plate was coated with 100 μL biotin-conjugated primary antibody against CXCL1 prior to incubation at 37 °C for 60 min. After three rounds of rinsing, the plate was coated with 100 μL of working fluid and further incubated at 37 °C for 30 min. Upon formation of the immunocomplex, the plate was washed five times, and 100 μL of TMB developer solution was placed into each well for colorimetric determination. After the reaction at 37 °C for 20 min, the absorbance of each well was immediately measured at 450 nm using a spectrophotometer . The level of the CXCL1 protein in the culture supernatant was quantified and analyzed based on a standard curve constructed using the absorbance values of the CXCL1 standard protein.3 cells per well were seeded into each well of 96-well plates supplemented with 100 µL medium containing various concentrations of human recombinant CXCL1 . (II) Autocrine effect: The same number of HeLa cells overexpressing CXCL1 and their mock controls, as described in experiment I, were grown in 96-well plates. One hundred microliter of medium supplemented with or without the ERK1/2 inhibitor, PD98059 , was added to each well. (III) Paracrine effect: First, the conditioned medium (CM) was prepared as follows: CXCL1-overexpressing PHM1-41 cells and their mock controls were inoculated in 100-mm diameter dishes at a density of 5 × 106 cells per 1000 µL of medium. After incubation at 37 ˚C for 24 h, the culture supernatant was harvested by centrifugation, and CM from PHM1-41 cells was produced by adding different proportions of the supernatant into complete medium. In the paracrine experiment, HeLa cells at a density of 2 × 103 cells per well were seeded into 96-well plates, and each well was incubated with 100 µL of different proportions of CM. The cells in the above three groups were incubated at 37 ˚C for 24 h, 48 h, and 72 h, and cell viability was determined using the Cell Counting Kit-8 (CCK-8) assay.Three separate experiments were used to evaluate HeLa cell viability. (I) Exogenous effect: HeLa cells at a density of 2 × 103 cells/well were seeded at the top chamber in 100 μL serum-free medium. The low chamber was coated with 600 μL serum-supplemented medium containing various concentrations of human recombinant CXCL1. (II) Autocrine effect: CXCL1-overexpressing HeLa cells (2 × 104) and their mock controls in 100 μL serum-free medium containing the ERK inhibitor, PD98059, or no inhibitor were seeded at the top chamber, whereas 600 μL medium with 10% FBS was placed at the low chamber for chemoattraction. (III) Paracrine effect: HeLa cells were maintained in the upper chamber as described in experiment I, and 600 μL medium containing various proportions of CM supplemented with 10% FBS was placed in the low chamber. Twenty-four hours after migration, the cells attached to the bottom surface of the Transwell filters were fixed and stained with methanol/crystal violet solution.Consistent with the proliferation assay, the following three experiments were carried out for the migration assay. (I) Exogenous effect: The same number of HeLa cells at a density of 2 × 10Total cell lysates were prepared from cell pellets using RIPA buffer  following the manufacturer’s protocol. After quantification of protein concentration, 40 µg of protein extracts was subjected to western blotting analysis. Different primary antibodies, including anti-p-ERK , anti- ERK , anti-cyclin D1 , anti-Bax , and anti-β‑actin , were utilized in the analysis. Thereafter, the samples were incubated with various HRP-conjugated secondary antibodies . Immune complexes were visualized using the Enhanced Chemiluminescence (ECL) Kit , and band intensities were calculated using the Gel imaging system .p-value < 0.05.All assays were carried out at least three times and SPSS 20.0 software was applied for statistical analyses. All values are expressed as the mean value and standard deviation. Statistical difference between two groups was determined using analysis of variance followed by the Student’s t-test. Statistically significance was defined as a To assess the clinical significance of CXCL1 in UCC, CXCL1 immunoreactivity was explored in uterine cervical tissue microarray. The results from the immunohistochemistry assay demonstrated that CXCL1 protein expression level was surprisingly increased in UCC tissues compared to non-cancerous tissues Fig. . In addiCervical cancer contains squamous cell carcinoma, adenocarcinoma, adenosquamous carcinoma, and other rare tumors, in which cervical squamous cell carcinoma (CESC) is the most common type of malignancy. We then used bioinformatic methods to study the CESC samples. The findings regarding the association between CXCL1 expression and patient survival probability are shown in Fig. Gene expression associated assay was used to identify the genes that were positively correlated with CXCL1 expression in CESC, and the genes with extremely low expression with median TPM (transcripts per milliom) < 0.5 were filtered out of the results. A total of 73 genes with pearson correlation coefficient (PCC) ≥ 0.30 were found  induced an evident dose-dependent increase in HeLa cell growth ability , granulocyte chemotactic peptide-2 (GCP-2/CXCL6), neutrophil-activating peptide-2 (NAP-2/CXCL7), interleukin-8 (IL-8/CXCL8), and growth-related oncogene (GRO) chemokines, that have three homologs , also defined as CXCL1, CXCL2, and CXCL3, respectively . The bioIn the current study, we demonstrated that the expression level of CXCL1 in UCC tissues was markedly elevated, and its higher expression was extremely associated with poor clinical stages. Consistently, it has been suggested that CXCL1 expression was frequently upregulated in several human tumors, such as melanoma, and breast and bladder cancers, and increased expression pattern of CXCL1 was predominantly relevant to several clinicopathological features, including high grade, advanced stage, and positive invasion in bladder cancer in vivo –30.Many studies have highlighted the crucial roles of CXCL1 in cancer development and progression. For example, secreted CXCL1 or CXCL1 overexpression in bladder cancer and melanoma was revealed to be correlated with the enhanced proliferative and metastatic properties of cancer cells, and CXCL1 in vitro was sufficient for cancer cells to generate tumors in a nude mouse xenograft model , 30. To +Gr1+ myeloid stromal cell population to the tumor microenvironment, which in turn generates several chemokine members that can serve as paracrine stimulators for the survival of tumor cells [Generally, it is agreed that the tumor microenvironment is composed of cancer cells, extracellular matrix, and different types of stromal cell subsets, which contain fibroblasts, vessel cells , and diverse inflammatory leukocytes  . To dateor cells . In the or cells .In the current study, we finally revealed that CXCL1 is a crucial chemokine that enhances UCC development by regulating extracellular signal-regulated kinase (ERK) signal-related genes, including ERK, p-ERK, cyclin D1, and Bax. ERK signaling is an essential transduction pathway that has received considerable attention because of its important effects in the regulation of various physiological and pathological processes. The activation of the ERK signaling pathway may stimulate tumor cell proliferation through its ability to initiate the expression of crucial growth-stimulating genes, such as cyclin D1 and early growth response-1 (EGR-1) , 35. TheIn summary, the current study demonstrated that CXCL1 expression was increased in tissues with UCC, and its higher expression was predominantly correlated with poor clinical stages. In vitro, CXCL1 may contribute the UCC cell malignant behaviors through exogenous, autocrine and paracrine manners, suggesting CXCL1 may be a potential diagnostic marker or therapeutic target in UCC.Additional file 1.


Anesthesia with sevoflurane resulted in the lowest serum peak levels of lipase and CRP during the first 3 days after transplantation, followed by desflurane and isoflurane . There was no difference with regard to 10-year pancreas graft survival as well as endocrine/metabolic function among all three VA groups. Multivariate analysis revealed the choice of VAs as an independent prognostic factor for graft failure three months after SPKT . Conclusions: In our study, sevoflurane and desflurane were associated with significantly increased early graft survival as well as decreased IRI-associated post-transplant clinical outcomes when compared with the isoflurane group and should be the focus of future clinical studies evaluating the positive effects of different VA agents in patients receiving SPKT.Background: Despite recent advances in surgical procedures and immunosuppressive regimes, early pancreatic graft dysfunction, mainly specified as ischemia–reperfusion injury (IRI)—Remains a common cause of pancreas graft failure with potentially worse outcomes in simultaneous pancreas-kidney transplantation (SPKT). Anesthetic conditioning is a widely described strategy to attenuate IRI and facilitate graft protection. Here, we investigate the effects of different volatile anesthetics (VAs) on early IRI-associated posttransplant clinical outcomes as well as graft function and outcome in SPKT recipients. Methods: Medical data of 105 patients undergoing SPKT between 1998–2018 were retrospectively analyzed and stratified according to the used VAs. The primary study endpoint was the association and effect of VAs on pancreas allograft failure following SPKT; secondary endpoint analyses included “IRI- associated posttransplant clinical outcome” as well as long-term graft function and outcome. Additionally, peak serum levels of C-reactive protein (CRP) and lipase during the first 72 h after SPKT were determined and used as further markers for “pancreatic IRI” and graft injury. Typical clinicopathological characteristics and postoperative outcomes such as early graft outcome and long-term function were analyzed. Results: Of the 105 included patients in this study three VAs were used: isoflurane , sevoflurane , and desflurane . Donor and recipient characteristics were comparable between both groups. Early graft loss within 3 months (24% versus 5% versus 8%,  Ischemia–reperfusion injury (IRI) is a major cause contributing to early graft injury, which ultimately results in severe patient morbidity and mortality in standard solid organ transplantation . HoweverPrevious studies have demonstrated the beneficial effects of volatile anesthetics (VA) in alleviating reperfusion injury in kidney and liver transplantation. Furthermore, transplant data are in line with evidence gained in general cardiovascular and neurosurgical procedures, which involve comparable phases of organ ischemia and reperfusion ,5,6,7,8.The pancreatic allograft is exquisitely susceptible to injuries in the early reperfusion period, which, among other serious complications, leads to damaging effects on the grafts’ microvasculature as well as dysfunction in both the endocrine and exocrine pancreas ,10,11. IOne interesting possibility that may attenuate IRI by inducing biochemical changes in different tissues seems to be pharmacological conditioning ,7. HereiThe aim of this study was to evaluate the effects of different VA regimes on early IRI-associated post-transplant clinical outcomes as well as on short- and long-term allograft function, survival, and (endocrine and metabolic) outcomes in patients who underwent SPKT.Medical data from all adult patients who underwent SPKT at the University Hospital of Leipzig between 1998 and 2017 were retrospectively analyzed. Our data source compromised a prospectively collected electronic clinical database. In this study, the main focus was placed on the anesthesia and perioperative protocols, as well as the early and long-term allograft function and patient outcomes. The original anesthesia perioperative and operative records were used to determine the usage of the applied anesthetic agents, as well as the perioperative anesthesiological management. Patients younger than 18 years, those receiving kidney transplantation alone (KTA), those receiving pancreatic re-transplantation, and those patients with insufficient/missing data about the perioperative, intraoperative, and postoperative anesthesiological status and outcome were excluded from the study.Pretransplant standard characteristics of the study population included recipient and donor parameters such as age, sex, body mass index (BMI), donor causes of death, and donor’s comorbidities and clinical course ). Further, recipient data comprised the duration of diabetes mellitus, time on the waiting list, the duration of pretransplantation dialysis, metabolic endocrine and lipid metabolism, and information on special comorbidities , blood pressure parameters and arterial hypertension, as well as the number of antihypertensive agents). Peri- and post-transplant data included information on operative and postoperative clinical course, including operation time, blood loss, cold and warm ischemia time of the pancreas and the kidney graft, administered amount and type of intraoperative fluids, amount and type of used catecholamine, and total volumes of blood product transfusions (fresh frozen plasma (FFP), red blood cells).The occurrence of “clinical IRI” was evaluated, and for the pancreas graft, it was defined as the development of clinically associated outcome parameters and consequences of pancreatic ischemia reperfusion injury, including graft pancreatitis, pancreatic abscess/peritonitis, early delayed graft function, graft thrombosis, rejection, and the consecutive need for re-laparotomy due to graft-related complications within 3 months. In this context, as described previously, the peak of C-reactive protein  and pancreas-specific serum lipase (mmol/L), which were defined as the highest serum levels within the first three days after transplantation, were used as further potential serological pancreatic IRI and graft injury markers . SecondaFurther, immunological and immunosuppressive characteristics (human leukocyte antigen (HLA)- mismatches, cytomegalovirus (CMV)- state, induction therapy), as well as patient outcome and long-term graft function and outcome, were analyzed.A further focus was placed additionally on the evolution and analysis of cardiovascular events following the SPKT. These included ischemic heart disease documented on a stress test or coronary angiography with or without the need for revascularization. Cerebrovascular accident (CVA) was recorded according to the presence of ischemic or hemorrhagic episodes. Further, peripheral vascular disease (PVD) events were assumed when revascularization or amputation was needed.Parameters of the endocrine, as well as lipid metabolism low-density lipoprotein (LDL)-cholesterol, high-density lipoprotein (HDL)-cholesterol ratio, HbA1C (%), C-peptide (ng/mL), and renal function variables (Creatinine (mmol/L) and urea (mmol/L)), were analyzed up to five years following transplantation.Acute rejection episodes were suspected if there were an abrupt increase in serum amylase/lipase or serum glucose levels, together with a significant drop in serum C-peptide level or increased serum creatinine levels and missing diuresis as well as abdominal pain associated with sonographic swelling of the graft. If possible, the diagnosis was confirmed from endoscopic biopsies of the duodenal segment of the graft. Biopsies of the kidney graft were performed to confirm a rejection. Pancreatic biopsies were not performed. Treatment of acute cellular rejection consisted of pulsed steroids or administration of antithymocyte globulin  in parallel with increased baseline immunosuppression. DGF of the kidney was defined as the requirement of dialysis in the first week following transplantation .All patients were administered general anesthesia via tracheal intubation with invasive mechanical ventilation. General anesthesia was induced with intravenous induction started with an opioid (sufentanyl or fentanyl) after routine preoxygenation, followed by an intravenous anesthetic  as well as the application of a muscle relaxant (rocuronium). General anesthesia was maintained with a halogenated agent  in an oxygen–air mixture and intermittent application of opioid and muscle relaxants according to the clinical evaluation of the anesthesiologist.Isoflurane, sevoflurane, and desflurane were used consistently throughout the study period as an individual choice made by the anesthetist.Additionally, to standard hemodynamic and ventilatory monitoring, direct arterial pressure and central venous pressure were continuously monitored. The arterial catheter was inserted into the radial artery, and the central venous catheter was placed into the right internal jugular vein.Our center protocol consisted of providing volume repletion and use of vasopressors, as needed, to achieve optimal and appropriate blood pressure at the time of graft reperfusion, ideally a systolic blood pressure level of >140 mmHg/MAP of >70 mmHg. In this context, the volume of fluids administered included crystalloids (mL), fresh-frozen plasma (FFP), human albumin, and transfusion of erythrocyte concentrate. In the case of MAP <70 mmHg, unresponsive to volume repletion, different types of catecholamine were used to maintain the targeted arterial blood pressure levels.As described previously, pancreas and kidney grafts were procured and transplanted following the international standards and guidelines provided by Eurotransplant ,31,32,33®) was used. Maintenance therapy included calcineurin inhibitors , and antimetabolites (Sirolimus (Rapamune®), Mycofenolate Mofetil (MMF); , and tapered steroids (Prednisolone®).The immunosuppressive protocol consisted of an induction therapy followed by triple maintenance therapy as described previously ,34. Short-test, χ2, analysis of variance (ANOVA), Kruskal–Wallis and Wilcoxon–Mann–Whitney test.With regard to baseline data, continuous variables are illustrated as mean values with standard deviation, and categorical variables are presented as whole numbers and percentages (%). For comparison between the study groups, the appropriate statistical significance test was used, including Student’s The primary endpoint of our study was the association and effect of VAs on pancreas allograft failure/survival following SPKT. In this context, pancreas graft failure was defined as resumed insulin therapy, removed pancreas, or re-transplantation. Kidney graft failure was defined as the need for dialysis, removed kidney, or re-transplantation. The secondary endpoint included “IRI-associated post-transplant clinical outcome” as well as long-term graft function and outcome.A stepwise Cox proportional hazard regression model was used to calculate hazard ratios (HR) with 95% confidence intervals (CI) for assessing pancreas graft failure/survival.p < 0.05 in univariate analysis.For multivariate analysis, we used a backward regression model including clinically relevant variables and those presenting Survival rates were calculated using Kaplan–Meier method, and a log-rank test was used to test statistical significance between groups. According to previous definitions, graft survival was calculated as the time from initial transplant to graft failure, censoring for death with a functioning graft and grafts still functioning at the time of analysis. Patient survival was defined as the time from transplant to patient death, censoring for patients still alive at the time of analysis. If a recipient was alive or lost to follow-up at the time of the last contact, survival time was censored at the time of the last contact.p-value <0.05 was considered statistically significant.All statistical analyses were performed using SPSS software . A “Note: This study has partially analysed data of a prospectively collected database with informations of transplant candidates and transplant recipients after pancreas transplantation. In parts, it has been previously published ,35,36,37p = 0.03) as well as the number of antihypertensive medications (p = 0.03) were recorded. Besides these, no other significant baseline recipient and donor characteristics were observed.During the study period, a total of 105 SPKTs were performed. Of those, 58 patients (55%) received isoflurane, 22 patients (21%) received sevoflurane, and 25 patients (24%) received desflurane as a VA, respectively. The mean follow-up period of the study was 151 ± 34.4 months. Donor and recipients’ demographic and clinicopathologic baseline characteristics according to the three volatile agents used in our study are illustrated in Intraoperative outcome parameters of the study group with regard to the VA’s used were described in p = 0.04), as well as rates of vascular thrombosis of the pancreas , occurred more frequently in the isoflurane group. In addition, there were significant differences in increased early pancreas graft loss within 90 days in the isoflurane group . No differences were observed with regard to renal graft injury stated as delayed graft function (p = 0.271) and rejection episodes (p = 0.228) between the three VA groups.The IRI-associated clinical postoperative outcome, as well as graft function outcome parameters following SPKT stratified by used VA, are shown in p = 0.001 for CRP and p = 0.039 for lipase).Further, recipients who received isoflurane had significant higher peak serum CRP and lipase levels  compared with the sevoflurane  and desflurane group . One-, three-, five- and ten-year pancreas graft survival rates in patients after SPKT were 76%, 73.9%, 70%, and 67.1% in the isoflurane group, 90.4%, 85.8%, 78.7, and 78.7% in the sevoflurane group, and 92%, 87.4%, 82.3%, and 75.9% in the desflurane group, respectively (log-rank test: p = 0.135 at 10 years) (p = 0.746). Overall long-term patient survival at 1, 3, 5, and 10 years was 90%, 87.9%, 83.9%, and 81.5% in the isoflurane group, 95.5%, 95.5%, 89.8%, and 89.8% in the sevoflurane group, and 92%, 92%, 87%, and 76.7% in the desflurane group, respectively (p = 0.581).Pancreas graft survival was significantly higher in the sevoflurane and desflurane group compared with the isoflurane group. During the first three months after SPKT, the percentage of pancreas graft loss was 24% in the isoflurane group, 5% in the sevoflurane group, and 8% in the desflurane group (log-rank test: 0 years) . One-, tWith regard to cox regression analysis, it could be shown that donor and recipient age, recipient BMI, and the duration of pancreas CIT are independent prognostic predictors of pancreas allograft failure within three months and five years after SPKT. Whereas the choice of VA, recipient gender, donor BMI and gender, transplant era, as well as the order of graft implantation showed a significant impact on pancreas graft survival at three months only, they had no significant effect on the 5-year graft survival .Previous experimental and clinical studies in various organs could demonstrate that anesthetic conditioning, specifically the use of VA, has pleiotropic effects capable of interfering with various pathophysiological pathways that mediate IRI in solid organ transplantation.Results from our study, which investigated the effects of three different VAs during general anesthesia in patients receiving a simultaneous pancreas and kidney transplantation (SPKT), are in line with previous findings obtained in the kidney ,7,38, liIn the course of SPKT, a number of potentially harmful processes inevitably occur, which affect both the viability of the allografts and, subsequently, morbidity and mortality of the graft recipient. Especially in pancreas transplantation, reperfusion injury reflected in elevated post-transplant serum lipase and CRP levels results in severe post-transplant pancreatitis, which consecutively might result in increased rates of graft vein thrombosis, both of which were observed in our isoflurane study group ,40. TranConditioning is a broad term generally used to describe strategies to attenuate IRI by inducing biochemical changes within the recipient and transplant allograft. Depending on timing and application, it can be referred to as pre-, peri-, and postconditioning. In this context, ischemic preconditioning and remote ischemic preconditioning have shown beneficial effects in solid organ transplantation, including SPKT . This deIn addition, several pharmacological substances were found to confer tissue tolerance to ischemia by underlying mechanisms similar to those mediating ischemic conditioning. This is also true for several anesthetics, accordingly termed anesthetic conditioning. This is particularly attributed to the volatile anesthetics (VA) used in our study and, to a lesser extent, to intravenous anesthetic agents ,42. WithHowever, CRP constitutes a rather unspecific marker of inflammation and infection, which may be elevated due to various infectiological reasons after SPKT and may therefore not be interpreted as a specific marker of IRI or indicator of pancreatic tissue injury. On the other hand, in the early phase, hence in the first 3 days after solid organ transplantation, infections are not very common and therefore do not represent a frequent cause of increased inflammatory parameters. In contrast, post-transplantation pancreatitis is known to cause an increase in CRP during the first few days after SPKT . TherefoIn previous studies, it could be demonstrated that peak CRP levels correlate well with IRI and the consecutive impairment of microcirculation in the early reperfusion period and that an elevation of CRP in the early phase after SPKT is associated with increased pancreas graft-associated complications . Based oWe would like to highlight the findings in this study of the protective effect of sevoflurane and desflurane compared with isoflurane during general anesthesia in SPKT, which may be of even more direct clinical importance. The use of volatile anesthetics during general anesthesia was associated with reduced IRI damage in various previous studies ,5,6,7,8.Several limitations of our study are important to discuss. Firstly, the low number of patients in each group and the retrospective nonrandomized design of our current study must be considered. Because of its retrospective design and the rather small number of patients included in the analyses, specifically per analysis group, the results of the study should be interpreted with caution. While direct translation into clinical practice is self-evidently prohibited due to potentially low statistical power and lack of shown causal relation, the purpose and value of retrospective trials are primarily the generation of hypotheses that need to be tested in prospective trials afterward.Secondly, although this study represents the results of a large German pancreas transplant center with equivalent surgical procedures and compact and robust follow-up data, the long investigation period, as well as different anesthesiologic and operative teams and styles, may have had an impact on the therapeutical decisions, for instance, preferred choice and used VAs in the previous years or different surgical techniques and the order of graft implantation.Hence, these variables  could be confounding in a multivariate regression analysis and may also introduce a bias into the observed outcomes in our study.Therefore, future research should focus on the effects of volatile anesthetics in SPKT and evaluate different dosages of VAs or varying timepoints during the perioperative procedure, as well as undergoing the same surgical graft implantation technique, combined with methods of recipient RIP, over a continuous stable time period.Alternatively, an interesting strategy for the prevention of IRI may be the evaluation of the effects of the application of different VA dosages and timepoints of exposure of the deceased organ donor to determine the effects of VAs before the onset of ischemia in IRI protection. Unfortunately, in our current study, retrospective data regarding the use of VAs in organ donors could not be obtained.Moreover, the use of intravenous anesthesia  may also imply a protective effect against oxidative stress and ischemia–reperfusion injury in major organs. Thus, total intravenous anesthesia is now being popularized in donor and recipient conditioning, with movement away from the VAs at some centers. First insights in the transplant setting with a small patient number and the combination of both agents have already recently shown promising organ protective effects with reduced IRI-associated complications as well as better graft function and outcome in living kidney transplantation . HoweverSevoflurane and desflurane were associated with significantly increased early graft survival as well as decreased IRI-associated post-transplant clinical outcomes when compared with the isoflurane group.In our current retrospective study, we found an association between the use of different volatile anesthetics during SPKT and amelioration of IRI with reduced post-transplant morbidity and graft failure. Hereby, we observed a significantly increased early graft survival as well as decreased IRI-associated clinical complications (mainly pancreatitis and graft thrombosis) in the sevoflurane and desflurane groups compared with the isoflurane group. On the other hand, no association between the applied volatile anesthetic and late graft survival and endocrine/metabolic function was observed. Future research in this area with the implementation of prospective, randomized clinical studies should thus focus on the potentially positive effects of different VA agents in patients receiving SPKT and on the investigation of the etiology and underlying cellular mechanisms of VAs and IRI as well as early and long-term graft outcomes. Therefore, administration of VAs at different timepoints and dosages in the deceased organ donor, or administration of the VAs in different dosages and in combinations with intravenous agents as well as in combination with RIP in the recipients during anesthesia for transplantation, might be of great interest.


In multivariable analyses, RPN was independently associated with fewer early post-operative complications than OPN  and shorter hospital stays . RPN was also a significantly associated with a decresedrisk of post-operative acute renal failure, and new-onset chronic kidney disease at 3 and 12 months post-surgery. There were no between-group differences in oncological outcomes. In comparison with OPN, RPN was associated with improved peri- and post-operative morbidity, better functional outcomes, and shorter hospital stays. Our results support the use of RPN, even for large and complex tumours.We compared the outcomes of robotic-assisted partial nephrectomy (RPN) and open partial nephrectomy (OPN) using contemporary data to respond to unmet clinical needs. Data from patients included in the registry who underwent partial nephrectomy between January 01, 2014 and June 30, 2017 within 20 centres of the French Network for Research on Kidney Cancer UroCCR were collected (NCT03293563). Statistical methods included adjusted multivariable analyses. Rates of peri- and post-operative transfusion, and of surgical revision, were lower in the RPN (n = 1434) than the OPN (n = 571) group (2.9% vs. 6.0%,  When technically possible, PN is also the standard of care in case of imperative surgery 1, and may be preferred in case of localised T1b and T2 tumours, depending on their complexity2. Recent data have shown that PN might also represent a feasible option for large kidney tumours (> 7 cm), with substantial but acceptable morbidity, limited risk of local recurrence, and excellent preservation of renal function3.According to the guidelines of the French and European associations of urology, partial nephrectomy (PN) is the standard of care for localised kidney T1a (≤ 4 cm) tumours4. However, the introduction of robotic assistance bypassed these limitations and led to an increasing proportion of renal cell carcinoma surgery being performed using minimally invasive procedures. In France, Ouzaid et al.5 reported that the annual number of robotic-assisted partial nephrectomy (RPN) procedures continuously increased between 2009 and 2015, whereas the number of laparoscopic partial nephrectomy (LPN) procedures remained stable over this period. Several systematic reviews and meta-analyses have highlighted the benefits of RPN compared to open partial nephrectomy (OPN)11 and LPN14. According to Autorino and Porpiglia15, RPN is becoming “the new gold standard for nephron sparing surgery” (NSS). However, the French health care system does not have a specific pricing system for this approach, and therefore does not reimburse the additional costs involved.The open approach has long been considered the gold standard for PN. Although the laparoscopic route was popularized in the 2000s, technical difficulties  limited its development16. Thus, in this article we report on the comparison between OPN and RPN outcomes.Data from the literature are usually based on retrospective series involving single centres and thus few of these data are contemporary and truly representative of French practices. The aim of our study was to use contemporary data, prospectively collected within the French Network for Research on Kidney Cancer UroCCR, to compare short- and mid-term PN outcomes according to the surgical approach. The recent report of the French National Authority for Health (HAS) on robotic-assisted nephrectomy indicated that RPN should be compared to OPN rather than LPN, due to the technical difficulties associated with the LPN approachCONTEMPORARI-PN  was a French, retrospective, observational, multicentre, cohort study based on the analysis of prospectively collected data from the UroCCR database . Informed consent was obtained from all participants. Collected data included patient and tumour characteristics, as well as surgery-related information. Partial nephrectomy could have been performed through OPN, LPN, or RPN. Thus, two-by-two comparisons were performed using a statistical model integrating these three arms: OPN, LPN, and RPN. However, only the comparisons between RPN and OPN are presented here.® Surgical System .All patients iwho underwent PN for kidney cancer and consented to be included in the database between January 01, 2014 and June 30, 2017 within 20 centres belonging to the UroCCR network were included into analyses. The surgical approach was chosen at the surgeon’s discretion. All procedures were performed by experienced surgical teams. RPN was performed using the da Vinci17) and late post-operative complications occurring within 30 days.The peri-operative morbidity of OPN and RPN was evaluated by measuring the rates of early post-operative surgical complications /creatinaemia before surgery) and GFR relative change ((GFR after surgery–GFR before surgery)/GFR before surgery) that could be more clinically significant than new-onset chronic kidney disease.Functional outcomes were evaluated before surgery and post-operatively on days 1–3 (D1–D3), and at month 3 (M3) and M12 by measuring the following parameters: creatinine, glomerular filtration rate (GFR) calculated according to the modified diet and renal disease (MDRD) formula, new-onset chronic kidney disease (CKD) for patients with pre-operative stage I or II CKD defined by a eGFR < 60 mL/min/1.73 m18  were also assessed.Length of hospital stay, and Trifecta achievement according to the Khalifeh criteria19 was not included in the model because variations in tumour size had already been taken into account as a prognostic factor. Stratified analyses were performed according to tumour size  and NSS indication. Rates of peri-operative transfusion, surgical re-operation and post-operative deaths within 30 days were compared between groups using the Chi-squared or Fisher exact tests, depending on the event distribution. The type I error rate was set at 5% for all comparisons. No missing data management strategy was used.Statistical analyses were carried out using SAS software version 9.3 . Qualitative variables were expressed as numbers and percentages. Quantitative variables were expressed as means and standard deviations. Early and late post-operative surgical complications, qualitative functional outcomes and Trifecta were compared between groups using mixed-effects logistic regression models, with the study centre as a random effect. Local and contralateral recurrence, and metastatic progression were described using the cumulative incidences approach to take into account the competing risk of death occurring before observing these events. Recurrence-free and overall survival were described using the Kaplan–Meier method. Oncological outcomes were compared between groups using stratified (according to study centre) Cox proportional-hazards models. Quantitative functional outcomes and length of hospital stay were compared between groups using mixed linear regression models, with the study centre as a random effect. Model assumptions were systematically verified. Quantitative functional outcomes and length of hospital stay were transformed into natural logarithms to comply with model assumptions. Removal of the random effect (for logistic and linear regression models) and/or use of the Firth’s penalization method (for Cox and logistic models) was proposed when models did not converge. All comparisons were done with and without adjustment for the following prognostic factors: tumour size, imperative NSS indication, age at surgery, American Society of Anesthesiologists (ASA) score, body mass index (BMI) at diagnosis and pre-operative creatininemia for post-operative acute renal failure and new onset CKD only. The RENAL scoreCommission Nationale de l'Informatique et des Libertés, CNIL): authorization DR 2013-206 and DR 2014-251. Moreover, the UroCCR project and this particular study (NCT03293563) were IRB-approved (Comité de Protection des Personnes (CPP) Sud-Ouest et Outre mer III, decision DC 2012/108). All patients gave their written consent before inclusion in the UroCCR database, after having received oral and written information about the UroCCR project.This study was performed in line with the principles of the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. In accordance with French law, the processing of personal data was approved by the French data protection agency  underwent either OPN (N = 560) or RPN (N = 1409). Patient and tumour characteristics, as well as surgical outcomes are presented in Table p < 0.0001) and severe  early post-operative complications compared with OPN , whereas no between-group differences were observed for late post-operative complications (4.6% vs. 6.5%). Multivariable analyses showed that RPN was independently associated with a decrease in both overall , and no significant difference was observed between groups (Table p = 0.0879), and stratified analyses showed that Trifecta outcomes were more likely to be fulfilled with RPN for tumours > 7 cm , the observed differences between RPN and OPN groups may not have been uniquely related to the surgery: there may have been differences at several levels of the patient care pathway including anaesthesia, management in the intensive care unit, and other post-operative parameters. Additionally, this previous French study20 included RPN performed by surgeons with various levels of expertise, with most of them still being on the RPN learning curve. This was not the case in our study.Our study responds to unmet clinical needs, and addresses the concerns of the HAS surrounding the lack of prospective contemporary studies on RPN21, except for estimated GFR outcomes, which were reported as better in patients who underwent RPN in two systematic reviews and meta-analyses11. Creatininaemia changes, post-operative acute renal failure, and new-onset CKD could not be compared with the literature due to a lack of published data.Our contemporary results are in line with those reported by several systematic reviews and/or meta-analyses11 and in a single centre study (RENAL score > 9)22, with reported benefits including reductions in blood loss and length of stay, as well as in intra-operative complications and transfusion rates22, and post-operative complications22. In contrast, apart from a shorter hospital stay after RPN versus OPN, Zargar et al. did not identify any between-group differences in their preliminary study focusing on solitary kidney23. To the best of our knowledge, the benefits of RPN for achieving Trifecta for large and complex tumours have not been demonstrated previously.Multivariable analyses showed that the benefit of RPN over OPN regarding early post-operative complications , post-operative acute renal failure, and new-onset CKD at M3 and M12 post-surgery was independent of the tumour size, imperative NSS indication, age at surgery, ASA score, and BMI at diagnosis. Interestingly, the superiority of RPN over OPN on Trifecta (which included zero peri-operative complications) was observed for tumours > 7 cm in stratified analyses. RPN therefore had a beneficial impact on these parameters, even for large and complex tumours. The seemingly greater benefit of RPN observed for new-onset CKD at M3 post-surgery for tumours > 7 cm may be related to the higher probability of occurrence of this event in patients with large tumours. The benefits of RPN over OPN for complex tumours have also been highlighted in a systematic review (RENAL score ≥ 7)21. However, the implementation of such RCTs is no longer conceivable due to the rapid expansion of RPN, and ethical issues. In France, some centres already only use the robotic approach for PN, convinced of the net benefits of this surgical approach, both for patients and hospitals. Outpatient RPN has even been implemented in Bordeaux for selected patients, supported by a dedicated, nurse-led clinical pathway, and has been shown to provide a high level of patient satisfaction as well as economic optimization of robotic assistance25.Several authors concluded their systematic review and meta-analysis on PN by claiming that randomized controlled trials (RCTs) to compare RPN and OPN would be needed to confirm their findings26. These advantages could balance out the cost of robotic assistance. In their meta-analysis, Wu et al.6 found that the overall cost of RPN was not significantly higher than that of OPN, and Bernhard et al.25 recently reported that implementing RPN along with enhanced recovery after surgery and day-case nurse-led protocols may facilitate the economic sustainability of robotic assistance for hospitals where the extra cost is not covered by the healthcare system.Our study had some limitations. Two-by-two comparisons were performed using a statistical model integrating an initial three study arms: OPN, LPN, and RPN. However, the results of the multivariable and stratified analyses remained stable after removal of the LPN arm, enabling us to report only on the clinically relevant comparison between OPN and RPN. The retrospective design of the study might lead to selection biases with more complex tumors and more imperative indications in the OPN arm. Although these differences were tackled with the multivariate analyses, other factors and unmeasured cofounders might affect the results and selection bias remains a concern. The exclusive practice of RPN in some of our study centres may have introduced a bias because no same-centre comparisons of outcomes were possible. However, our study was multicentric and gathered data from 20 centres, all of which were expert centres for both RPN and OPN. Furthermore, the impact of surgeons’ expertise on outcomes also needs to be considered. Although this is a factor that cannot be eliminated, our study, like most previous evaluations of RPN, used data generated by high-volume surgeons with extensive experience. Our findings may not therefore be readily transferable to settings were RPN is conducted by less experienced surgeons. Another question to be addressed is the comparative cost-effectiveness of the two surgical approaches. Our findings demonstrated that patients who underwent RPN had shorter hospital stays, fewer complications and re-operations than those who underwent OPN. Other previously reported advantages of minimally invasive PN over OPN that may also have some impact on costs include lower rates of peri-operative opioid use and fewer days of workplace absenteeismIn conclusion, RPN was associated with improved peri- and post-operative morbidity, shorter hospital stays, better functional outcomes for some parameters, and similar oncological outcomes in comparison with OPN in our multicentre contemporary study. Our results support the use of RPN, even for large and complex tumours.Supplementary Information 1.Supplementary Information 2.Supplementary Information 3.


Dear readers,In the year 2006, I have published with great enthusiasm my first paper in Virchows Archiv . Since tHaving that said, I am most thankful to the European Society of Pathology (ESP) and Springer for trusting me to overtake the challenging and demanding role of the new Editor-in-Chief of Virchows Archiv. With a mixed feeling of honor and privilege, enthusiastic commitment, and loyalty, I appreciate and admit, how difficult this task will be, when considering the great effort and the tremendous work the past editors of the journal since the union of its A and B sections have put forward, to insure such a successful development of our journal. The evolving future plans and the implemented great work, which became more visible during the editorship of Fred Bosman  and DaniCell Pathology). At same time, these studies should have a significant positive impact on the practice of our discipline in one of its three major pillars: diagnostics, prediction and prognosis, thereby contributing to the improvement in patient’s care, the ultimate goal of our job. Hence, a major feature of acceptable articles is that they should unequivocally positively influence the practice of modern diagnostic and predictive surgical pathology in line with the historical section A of the journal .During my Chief Editorship term, my goal will be to maintain the established line and high standard of Virchows Archiv, at same time trying to make the unification of the historical A and B sections of the journal more visible in each published article. Accordingly, the types of articles we wish to receive and publish should contribute to our understanding of the scientific basis and the complex mechanisms underlying human diseases including molecular and translational studies, so fulfilling the goals of the historical section B of the journal  diseases are much welcome.Besides the annual review issue, my aim will be to have at least one major review article published in each issue addressing timely hot topics related to Finally, no need saying, that the great success the journal has made and the anticipated further development of our journal would haven´t been and wouldn´t be possible without the tremendous joint efforts of our editorial board, the associate editors and the (much appreciated) demanding work by our reviewers; all being inevitable for maintaining the high-quality scientific contents which became the standard in each published issue of Virchows Archiv.Abbas Agaimy, M.D.Editor-in-Chief


Erratum zu:Schmerz 202210.1007/s00482-022-00671-9In diesem Artikel waren die Adressdaten für den Autor Lars Neeb falsch angegeben. Statt „Neurologische Klinik und Poliklinik, Institut für Public Health, Charité-Universitätsmedizin Berlin, Freie Universität Berlin und Humboldt Universität zu Berlin, Berlin, Deutschland“ hätten sie „Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt Universität zu Berlin, Klinik und Hochschulambulanz für Neurologie, Berlin, Deutschland“ lauten sollen.Der Originalbeitrag wurde korrigiert.


The starch granules were visualized by SHG imaging, while the chloroplasts in mesophyll cells were visualized by TPF imaging. Our results provided micron scale spatial resolution of starch distribution within leaves and showed starch circadian patterns consistent with those profiled by enzymatic assays in previous studies. We demonstrated that TPF-SHG imaging is a potential tool for revealing the real-time heterogeneity of starch circadian rhythm in leaf cells, without the need for destructive sample preparation.Plant chloroplasts conduct photosynthesis to convert solar energy into sugars for the carbon source essential for cell living and growth during the day. One fraction of photosynthetic products is stored in chloroplasts by forming starch granules to continue the provision of carbon energy during the night. Currently, profiling the starch temporal pattern requires either: (i) sacrificing the leaves, or (ii) generating transgenic plants at the risk of changing the metabolisms by incorporating a genetically modified granule-bound starch synthase (GBSS). In this paper, we demonstrated a nondestructive method using two-photon fluorescence (TPF) and second-harmonic generation (SHG) imaging to quantify starch granules within chloroplasts of fresh intact leaves across a day-night cycle. We did so using two Arabidopsis lines having normal and excess starch contents: wild-type (Columbia-0) and  There are two primary places where starch is produced in plants. One is in the leaves where photosynthesis takes place. The other is in the storage organs such as the stems and roots. The starch in the storage organs accumulates continuously and is not degraded until it is needed for growth. Starch in the leaves, however, has a diurnal variation. During the day, photosynthesis produces starch that accumulates in the chloroplast then forms into starch granules3. When photosynthesis ceases at night, the energy needed for cell survival and growth is provided by the degradation of starch stored in the leaves during the day6. The ability to monitor immediate starch variations is important for tracking diel mechanisms involved in starch metabolism.Starch is the main storage of carbohydrates in higher plants8, starch-iodine staining, and genetic modification. Enzymatic methods extract and degrade starch with enzymes and can be used to quantify the total amount of the glucose residue stored in starch9. The iodine solution staining method uses iodine to dye the starch without extracting it from the leaf. But to observe the stained starch clearly, the sugars and chlorophyll need to be dissolved and removed with alcohol. This method allows determination of the starch position and distribution in the tissue, but due to the requirement for dissolving the chlorophyll and staining with iodine solution, the method destroys the leaves during the preparation process. Another method for starch determination is by cloning a construct that contains a target gene of interest, then obtaining the transgenic plants that carry the construct randomly integrated into the genome. Using an enzyme that participates in starch metabolism and fusing it to a fluorescent protein results in a transgenic plant that allows the observation of starch through fluorescent imaging11. Although utilizing fluorescent protein imaging for quantitative starch analysis does not destroy the transgenic leaf, creating a transgenic plant is complicated and time-consuming, and needs to be done for each ecotype of interest. Another non-destructive method that had been used to quantify starch in plants is microCT imaging, which was applied specifically to the ray and axial parenchyma (RAP) in woody stems12. Because this last method does not provide unique contrast from starch, it relies on using the shapes and sizes of the RAP cells from image analysis, and cannot easily be applied generally for starch determination.Methods commonly used for starch quantification include enzymatic methods15, a possible method for its identification is by second harmonic generation (SHG)17. SHG is a nonlinear optical process that is allowed only in non-centrosymmetric structures and has been used as a contrast mechanism in multiphoton microscopy for imaging a variety of intact biological tissues such as bones, tendons, and cartilage20. It has also been shown to provide contrast for imaging starch and cellulose in plants 21–23. For SHG starch studies, research groups have mostly analyzed starch structure by using SHG detection after the starch had been extracted26. The SHG images acquired usually relied solely on the use of spectral filters to separate the SHG signals from the fluorescence. Without spectral confirmation, wide bandwidth fluorescence signals close to the SHG spectral band can leak through the SHG filter and be detected as SHG intensity, resulting in a misrepresented SHG image. In this study, we used SHG imaging to profile the starch content of intact Arabidopsis leaves throughout a 24-h light–dark cycle. SHG imaging in leaf cells was subcellularly colocalized with TPF signals emitted by chloroplasts, suggesting that the SHG imaging was generated by starch granules. We developed a method based on SHG and TPF intensity images to quantify the starch content inside the chloroplast. Furthermore, spectral mapping of the sample further confirmed the SHG images faithfully represented the SHG intensity variations. We demonstrated the effectiveness of our method to monitor nondestructively starch variations using two kinds of Arabidopsis: One wild-type Columbia-0 (Col-0) with normal starch production, and one sex1 mutant with a defective SEX1 that prevents starch degradation and utilization, resulting in an excess of starch accumulation in the chloroplast. We observed that the SHG imaging signal increased during the day and decreased during the subsequent night in Col-0 leaves but remained consistently high in sex1 leaves. The different variations of SHG under diel cycles between Col-0 and sex1 demonstrated that the fluctuation of starch granules in living cells can be effectively detected with SHG imaging.Since starch granules are semicrystallinesex1 leaves. From the TPF images, contrast from the chlorophyll autofluorescence allows visualization of chloroplasts in mesophyll cells. It can be seen that the autofluorescence does not completely fill the chloroplasts and there are many dark sites. The middle column shows the SHG images from the same scanning area, where the SHG signal is expected to come from the starch granules. In the third column, merged TPF and SHG images show that the SHG appears inside the chloroplasts, in the areas where the autofluorescence signal is weak. This is expected since starch granules occupy spaces in the stroma, while the chlorophyll is restricted at the thylakoid membranes, resulting in the spatial separation of the TPF and SHG signals. Furthermore, we found that SHG signals in the sex1 leaf are stronger than those in the Col-0 leaf, consistent with the excess starch known to be present in the sex1 mutant.Figure sex1 leaf, but with a shorter spectral integration time of 0.3 s. Due to the greater amount of starch present in the sex1 mutant, the shorter integration time can produce spectra with SHG intensities comparable to that of the Col-0 wild type. These spectral observations provide confirmation that the SHG filtered PMT images correspond to the SHG intensity, allowing us to use the SHG intensity to quantify the amount of starch present in the leaf. The greater SHG intensities observed in sex1 are consistent with the fact that sex1 mutants lead to greater starch accumulation in the chloroplast than the Col-0 wild types.To confirm that the PMT images acquired with the SHG and TPF bandpass filters correspond to the spatial distribution of starch SHG and chlorophyll TPF, we acquired a spectral mapping in a selected region of the PMT mapping. We first obtained the combined TPF and SHG PMT image of a fresh intact Col-0 leaf in the NDD configuration Fig. a. Then wsex1 leaves across a 16-h light/8-h dark cycle. The intensity values correspond to the detected photon count during the pixel integration intervals. We acquired images at time intervals of 3 h during the light–dark cycle and calculated the ratio of SHG intensity to TPF pixel count  were kindly provided by Jychian Chen from the Institute of Molecular Biology, Academia Sinica, Taipei28. All plant studies were performed in accordance with the relevant guidelines and regulations provided by the authors’ respective institutions and the Taiwan Centers for Disease Control. We first sowed the bleach-cleaned Col-0 or sex1-1 seeds on half-strength Murashige and Skoog (1/2 MS) with vitamins  in phytoagar plates, and placed the plates in a growth chamber . The seeds were then incubated in a long-day (LD) photoperiods of 16-h light/8-h dark at 22 °C for 7–24 days. Just prior to imaging, we removed the plate from the chamber at the specific Zeitgeber Time (ZT) and detached the largest leaf from the seedling. The leaf was then directly placed on a glass slide, kept moist with drops of water, and covered with cover glass for imaging. For the starch diurnal variation experiment, we put the plates with sowed seeds in two separate growth chambers , with the photoperiod initial time separated by 12 h. This allowed us to acquire data at ZT from 0 to 24 h in a half day period. We obtained images for ZT starting at 0 h when the light first came on and proceeded with 3 h intervals until ZT24.The seeds of wild type Columbia-0 (Col-0) and the 2. A 670/30 nm filter was used for imaging chlorophyll TPF and a 532/3 nm filter was used for SHG imaging. For hyperspectral imaging, the signal was collected in the DD configuration by a 200 μm core fiber, then sent to a fiber-coupled spectrometer , containing a 300 grooves/mm grating and a TE-cooled CCD .The TPF and SHG imaging were performed on a custom-built laser scanning system . This system allowed for SHG and TPF imaging in the non-descanned detection (NDD) configuration and hyperspectral imaging in the descanned confocal detection (DD) configuration. The laser source was a 300-fs pulsed laser with a repetition rate of 40 MHz and a central wavelength of 1064 nm . A 20X objective lens  focused the laser light with an on-sample power of 23–27 mW corresponding to a power intensity of 10 mJ/s‧μmSHG, to the TPF pixels count, CntTPF, above a threshold for each image data. The SHG intensity was summed over only those pixels with a SHG count above the background level of 2, for the integration time of 0.128 ms. For the TPF pixel count, we used a threshold level of 50. This value was determined empirically to best delineate the chloroplast morphology that is in the focal plane of the image (See Since the starch granules are produced in the chloroplast of the mesophyll cells, we quantified the starch content by calculating the ratio of SHG intensity, ISupplementary Information.


Centre for Evidence in Transplantation have developed the Transplant Trial Watch. The Transplant Trial Watch is a monthly overview of 10 new randomised controlled trials (RCTs) and systematic reviews. This page of Transplant International offers commentaries on methodological issues and clinical implications on two articles of particular interest from the CET Transplant Trial Watch monthly selection. For all high quality evidence in solid organ transplantation, visit the Transplant Library: www.transplantlibrary.com.To keep the transplantation community informed about recently published level 1 evidence in organ transplantation ESOT and the Cardiovascular Magnetic Resonance for Rejection Surveillance After Cardiac Transplantation.by Anthony, C., et al. Circulation 2022; 145(25): 1811–1824.The aim of this study was to investigate the feasibility of cardiovascular magnetic resonance (CMR)-based monitoring for cardiac allograft rejection.Participants were randomised to receive either CMR-based or endomyocardial biopsy (EMB)-based rejection surveillance.40 orthotopic heart transplant recipients.The primary endpoint was frequency and cumulative freedom from significant (>grade 2R) rejection. The secondary endpoints included frequency and cumulative freedom from low-grade (grade 1R) rejection, kidney function, hospitalisation, duration of hospital stay, infection, myocardial function, death, immunosuppression exposure and the incidence of biopsy-related complications.1 year.This is an interesting and well-conducted study in cardiac allotransplantation. Cardiac magnetic resonance (CMR) imaging was compared to the standard protocol of surveillance for rejection with the invasive endomyocardial biopsy (EMB). The first part of the study was a cross-sectional analysis to understand cut-off values for acute rejection and the second part was a randomised study comparing the two surveillance methods using this information. The trial was set up as a noninferiority study and therefore the inclusion did not need to be large (20 in each arm), as per prior power calculation. A detailed analysis of the CMR validation is provided. The primary endpoint was frequency of significant rejection (grade 2R or higher). This was found to be similar in the two groups. In order not to miss high grade rejection, treating physicians could request EMB on any patient at their discretion. This option was taken 11 times in the CMR group and in 9 of these cases the EMB result was identical to the CMR result. In this single-centre study, a surveillance protocol using CMR instead of EMB in cardiac allograft recipients was safe, feasible and offers significant advantages over invasive cardiac biopsies.3.Strict intention-to-treat analysis.Yes.ACTRN12618000672257.Non-industry funded.Hypothermic Oxygenated Perfusion in Extended Criteria Donor Liver Transplantation—A Randomized Clinical Trial.by Ravaioli, M., et al. American Journal of Transplantation [Online ahead of print].The aim of this study was to compare the effect of hypothermic oxygenated perfusion (HOPE) vs. static cold storage (SCS) in extended criteria donor (ECD) liver transplantation.Participants undergoing transplantation of an ECD liver graft were randomly assigned to receive a liver after HOPE or after SCS alone.135 potential ECD liver grafts were randomised, of which 110 were used for liver transplantation.The primary outcome was the incidence of early allograft dysfunction (EAD). The secondary outcome were patient survival, graft survival, the early allograft failure simplified estimation (EASE) risk score, and the rate of graft or other graft-related complications.1 year.This is an interesting and well-conducted trial in ECD liver transplantation. Livers were randomised to standard static cold storage (SCS) or to a period of Hypothermic Oxygenated Perfusion (HOPE), using the Vitasmart device . Organs in the HOPE group had a period of SCS of 4–5 h on average prior to starting HOPE for 2–3 h on average. No organ was discarded during perfusion. The study was single centre and designed with a prior power calculation to determine sample size. The primary endpoint was Early Allograft Dysfunction (EAD) using a well-established composite definition. There was a significant reduction in EAD with HOPE compared to SCS (13% vs. 35%) and also a significant reduction in re-transplantation (0% vs. 11%). This form of HOPE, using just portal vein perfusion in ECD liver transplantation, is associated with better early allograft function, which is very likely to impact on longer term function and graft survival.2.Per protocol analysis.No.ClinicalTrials.gov—NCT03837197.Non-industry funded.ex-vivo . The technologies differ in their simplicity/ease of use, ability to assess organ viability and the duration of safe perfusion.The shortage of suitable donors to meet demand has resulted in increasing use of extended criteria donor (ECD) organs to try to address the mismatch. ECD donor organs are known to be more at risk of adverse post-operative outcomes due to increased vulnerability to ischaemia-reperfusion injury. In attempts to counter this additional risk, there has been a great deal of interest in novel perfusion technologies to recondition, repair and assess grafts prior to transplant. Such technologies can be used in the donor  or In a recent paper in the American Journal of Transplantation, Ravaioli and others report a single centre randomised controlled trial of HOPE after static cold storage (SCS) versus SCS alone in ECD liver grafts . 110 recPerhaps the most striking finding is that graft survival was significantly higher in the HOPE arm of the study, a finding not seen in the larger multicentre studies of HOPE or NMP. A detailed breakdown of causes and timings of graft loss is not provided, making the role of perfusion in this finding difficult to interpret. Another interesting finding is the numerically lower incidence of acute rejection in HOPE livers. This has been seen previously with use of HOPE in kidney transplantation , and mayOverall, these findings support previous studies in both liver and kidney transplantation that HOPE is a safe, simple and effective method of preservation which may be beneficial in marginal donor organs.