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					program                    Malmö, Sweden, October 8–10 2004




     th
5
European
Transplant
Fellow
Workshop



             The ETFW is an official educational activity of the European
             Society for Organ Transplantation (ESOT) and organised under
                  the auspices of the Educational Committee of ESOT.
               Host ETFW 2004: Professor Henrik Ekberg, Malmö, Sweden.




                           Sponsored by:
  4 Scientific program
  6 Interactive presentations
  7 Interactive lectures
  8 Participants’ scientific presentations
  9   Social program
 10 The European Transplant Fellow Workshop
 12 Abstracts
 60   Contact details of participants
 62   Contact details of faculty members



contents
                                                                                      th
                                                                              5
                                                                              European
                                                                              Transplant
                                                                              Fellow
                                                                              Workshop




Venue                            Local organizing committee    Local sponsor person:
Börshuset, Conference center     Henrik Ekberg, MD, PhD        Marie Sverkersdotter
Skeppsbron 2, Malmö              Nils H Persson, MD, PhD       Fujisawa Scandinavia
(5 minutes walk from the hotel   Henrik Thorlacius, MD, PhD    Tel: +46 (0)708 711 575
Mäster Johan)                    Clara Påhlman, MD             m.sverkersdotter@fujisawa.se
                                 Helene Malm, MD
Hotel Accomodation                                             Travel information
Hotel Mäster Johan               Correspondence                All tickets and travel information
Mäster Johans gata 13            Prof Henrik Ekberg            will be send from Väsby Resebyrå
Malmö                            Dept of Nephrology and        Ms Karin Eriksson
Tel: +46 (0)40-664 64 00         Transplantation               Tel: +46 (0)8-590 720 50
Fax: +46 (0)40-664 64 01         Malmö University Hospital     karin.eriksson@vasbyrb.se
www.masterjohan.se               S-205 02 Malmö, Sweden
                                 henrik.ekberg@kir.mas.lu.se   Sponsor
                                                               Fujisawa
scientific program
                                                           Friday, October 8
= Interactive presentation based on cases
                                                           Afternoon session
= Presentation by the fellows
                                                           13.00 – 13.30
= Interactive lecture (30 min) + discussion (15 min)
                                                           Welcome and introduction
= Interactive lecture (45 min) + discussion (15 min)       of the fellows
                                                           Henrik Ekberg, Sweden
                                                           13.30 – 14.15
                                                           Donor selection; contra-indications
                                                           and expanded donor criteria.
                                                           Nils H Persson, Sweden
                                                           14.15 – 15.00
                                                           Ischemia/reperfusion injury
                                                           and long-term consequences
                                                           for graft outcome.
                                                           Robert Porte, Netherlands
                                                           15.00 – 15.30
                                                           Fruit and coffee
                                                           15.30 – 16.30
                                                           Clinical problems early after
                                                           kidney transplantation.
                                                           Johann Pratschke, Germany
                                                           16.30 – 18.00
                                                           (Abstract no. 1 – 6)
                                                           Chair: Robert Porte
                                                           Henrik Thorlacius
                                                           18.30
                                                           Bus departs from the Hotel

                                                           18.45–19.45
                                                           Sauna and Swimming at “Ribban”

                                                           20.00
                                                           Dinner at Restaurant
                                                           Salt & Brygga by the Sea




                                                       4
Saturday, October 9                Sunday, October 10
                                   Morning session

08.30 – 09.15                      08.30 – 09.30
Recent trends in initial immuno-   Clinical problems in liver
suppressive protocols              transplantation.
Henrik Ekberg, Sweden              Jan Lerut, Belgium
09.15 – 10.15                      09.30 – 10.15
Monitoring and toxicity            (Abstract no. 19 – 21)
of currently used immuno-          Chair: Dirk Kuypers
suppressive drugs.
Dirk Kuypers, Belgium              10.15 – 10.30
                                   Fruit and coffee
10.15 – 10.45
Fruit and coffee                   10.30 – 11.15
                                   Recent development in islet
10.45 – 11.45                      transplantation.
Clinical problems long-term        Olle Korsgren, Sweden
after kidney transplantation.
David Wheeler, UK                  11.15 – 12.30
                                   (Abstract no. 22 – 23)
11.45 – 12.30                      Chair: Olle Korsgren
(Abstract no. 7 – 9)               John Dark
Chair: Johann Pratschke
Nils H Persson                     12.30 – 12.45
                                   Conclusion
12.30 – 13.30                      Henrik Ekberg
Lunch
                                   13.00
13.30 – 14.30                      Lunch
Clinical problems in thoracic
transplantation
John Dark, UK

14.30 – 15.15
(Abstract no. 10 – 12)
Chair: David Wheeler

15.15 – 15.45
Fruit and coffee

15.45 – 17.15
(Abstract no. 13 – 18)
Chair: Jan Lerut
Henrik Ekberg

19.30
Dinner at Restaurant Årstiderna
(3 minutes walk from
Hotel Mäster Johan)




                                   5
interactive
presentations
based on cases and planned to
include the following aspects:



    1. kidney transplantation – early (surgery)
      ATN, ureteric stenosis, ureteric leakage, insufficient circulation,
      arterial thrombosis, lymphocele, infection, acute rejection


    2. kidney long-term (transplant nephrology)
      deterioration of graft function and long-term immunosuppression;
      CNI nephrotoxicity, recurrent disease, chronic transplant neph-
      ropathy; CMV- and BK virus infection, cardio-vascular disease,
      malignancy


    3. thoracic transplantation
      surgical complications, infection, acute rejection, graft dysfunction
      early and late, bronchiolitis obliterans syndrome


    4. liver transplantation
      hepatic artery thrombosis, biliary complications, infection, acute
      rejection, graft dysfunction early and late


    5. donor evaluation
      relative contra-indications for acceptance of deceased donor
      – such as old age, cardio-vascular disease, impairment of organ
      function, infection, lenght of ICU stay, laboratory abnormalities,
      virology




                                                 6
interactive
lectures
– not specifically based on cases,
but still interactive



    1. islet transplantation
      Lab and clinical techniques, practical difficulties – several donors,
      immunosuppression, clinical results


    2. recent trends in initial immunosuppressive
       protocols
      CNI-free, steroid-free protocols and for high-risk patients


    3. ischemia/reperfusion injury and long-term
       consequences for graft outcome
      Mechanisms, current principles of organ preservation, clinical
      implications


    4. drug monitoring and toxicity
                                   ,
      to cover CsA, tacrolimus, MMF sirolimus and steroids




                                             7
participants’
scientific
presentations
 The participants of the ETFW are selected based on submitted abstracts of research
 activities. The reason is that research activities is a fundamental part of the training
 to be a successful physician or surgeon in the field of transplantation. Presentation of
 research results is likewise very important – without a successful presentation, your
 research results will not reach out to your colleagues. Therefore presentation of scien-
 tific efforts is an important part of the workshop.


 It is easy to describe a research project when given 30 minutes to do so. It is much
 more difficult if given only 5 minutes. Only the most important aspects, the essence of
 the project may be summarised in this short time. It is similar to oral presentations of
 posters organised at some congresses – headline news only.


 Each participant has 5 minutes to give a talk supported by his slide presentation using
 Power Point software. In principle only 5–10 slides can be shown in this short time, the
 number depending on the simplicity (or complexity) of the slides. It is advicable to use
 simple slides, preferably only 5–10 lines of text on each slide. Perhaps 1–2 slides for
 Background, 1 for Aim of study, 1–2 for Methods, 1–3 for Results and 1 slide for Conclu-
 sions. No double projection of two slides back to back will be used.


 After each presentation we will have 10 minutes of discussion. There will be a faculty
 member leading this discussion and two selected participants are giving the first com-
 ments. In addition everybody else is ofcourse taking part. Main issues to discuss will
 include:


  1. What was the message of the presentation?
  2. Was the conclusion correctly based on the results?
  3. Was the presentation well done and clear?
  4. Any unnecessary or unclear slides?
  5. Clinical or experimental relevance of the results.

 This is most probably the way your tutor and colleagues would comment on your pres-
 entation when you are preparing to give a talk at an international congress. So look at
 this as a last rehersal before going abroad. Good luck.


 Henrik Ekberg




                                               8
social program
      Friday October 8

      18.30
      Bus departs from the Hotel
      18.45–19.45
      Sauna and Swimming at “Ribban”
      20.00
      Dinner at Restaurant Salt & Brygga by the Sea

      Saturday October 9

      19.30
      Dinner at Restaurant Årstiderna
      (3 minutes walk from Hotel Mäster Johan)




                  9
The
European
Transplant
Fellow
Workshop
Sponsored by Fujisawa.
Under auspices of the European
Society for Organ Transplantation (ESOT).
Concept and Protocol.



    Objectives
    To provide a podium for young clinicians, who are specializing in clinical organ trans-
    plantation, to discuss new and current topics in transplantation with established and
    internationally recognized leaders in this field and to provide an opportunity to meet
    peers and make contacts and networks that may enhance future collaborations and
    (multi-center) research.


    Format
    The workshop will be organized as a 3-days meeting. The program includes a mix of
    state-of-the-art lectures, case discussions and interactive courses by internationally
    recognized leaders in the field of transplantation and short presentations of original
    scientific work by the attending fellows.




                                                10
Attendees
The workshop will be open for about 20–25 young clinicians (<40 year) who are cur-
rently being trained in the field of organ transplantation, either as a physician or sur-
geon, or who are a junior staff member at one of the Transplant Centers in the Europe.
The number of attendees will be limited to enhance active participation and to stimu-
late communication between attendees and program faculty. Directors of Transplant
Programs in Europe will be invited to submit name and curriculum vitae of a person
they would like to propose as a candidate for participation in the workshop. Submis-
sion should be accompanied by a supporting letter from the program director or head
of the department. Based on their C.V and/or previous achievements it should be evi-
dent that candidates are pursuing a career in the field of clinical organ transplantation.
Participants must attend the entire workshop and are expected to actively participate in
the discussions.


Contributions by attendees
All attendees have to submit an abstract summarizing research work they have per-
formed in the field of transplantation or a unique clinical case report, which he/she will
be able to present at the workshop. Research work should contain original material
that has not yet been published or presented at an scientific meeting. Presentations
will be evaluated by the program faculty present at the Workshop and discussed with
the attendee after his/her presentation. Attention will be given to the scientific content
as well as to the presentation itself. Attending fellows will be selected based on their
C.V. and abstract submitted.


Award
Two awards will be given for the best presentations given by two of the attending fellows
at the workshop.


Program Faculty
Established and leading clinicians in the field of organ transplantation will be invited
to present interactive courses, case discussions or state-of-the-art lectures of about 45
minutes, covering various subjects of transplantation.


Scientific program
A scientific committee formed by members of the ESOT educational committee, the
EFTW chairman, and the local host. This scientific committee will be responsible for
the scientific program and the selection of the faculty presentations. Each year another
European transplant center will be invited to host the EFTW.


Role of Fujisawa
Fujisawa will collaborate closely together with ESOT, and will make this workshop
financially possible through an unrestricted educational grant. Costs for accommoda-
tion and traveling (economy class) of all attendees and the program faculty will be cov-
ered by Fujisawa. Representatives of Fujisawa may be present during the meeting, but
there will be no commercial or sales presentations.




                                          11
                                                       12




Detection of oxidative injury with or without
small bowel ischemic preconditioning prior
to autotransplantation
Andrea Ferencz, János Toldi, Zsolt Fehér, Elizabeth R th
University of Pécs, Medical School, Department of Surgical Research and Techniques, Hungary




          Aims
          It is well known that cold preservation prior to small bowel transplantation can moder-
          ate tissue injury, however this procedure is unable to protect the organ against acute
          reperfusion injury. The aims of our work was to follow up the oxidative stress param-
          eters during small bowel autotransplantation testing the protective effect of ischemic
          preconditioning (IPC) especially to investigate the role of transcriptional factor Nuclear
          Factor-kappaB (NF-kB) in the complex cascade of IPC.


          Methods
          In group I (GI) and group II (GII) total orthotopic intestinal autotransplantation was per-
          formed on 20 mongrel dogs. In GI and GII grafts were perfused by mesenteric artery
          and stored in cold University of Wisconsin (UW) solution. In GII before preservation IPC
          was induced via occlusion of the mesenteric artery with 4 cycles each of 5 minutes of
          ischemia and 10 minutes of reperfusion (IPC protocol). In GI and GII after 3 hours pres-
          ervation, 1-hour of reperfusion was allowed to assess graft viability. In group III (GIII)
          intestines were preconditioned with 4 cycles without preservation. To determine the
          oxidative stress in bowel tissue marker of lipidperoxidation malondialdehyde (MDA),
          reduced glutathione (GSH), and activity of superoxide dismutase (SOD) as endogenous
          antioxidants were measured. In GIII cytoplasmic (total) and nuclear (activated) NF-kB
          level were detected after laparotomy (control), 30 minutes, 1 hour and 2 hours follow-
          ing IPC by chemiluminescences based ELISA method.


          Results
          Our finding showed slightly elevated GSH concentration in GI, while SOD activity
          decreased significantly during reperfusion (control 264.28±10.61 IU/g; reperfused:
          103.28±15.49 IU/g, p<0.05). In GII GSH elevated markedly, and better preservation of
          SOD could be measured (reperfused: 189.12±16.36 IU/g). Our measurement in GIII
          revealed a specific dynamism of NF-kB activation. Total and activated form of NF-kB
          significantly elevated 30 minutes following IPC (p<0.05 vs. control). 1 hour after IPC NF-
          kB activation decreased to the control level, however 2 hours after IPC gradual activa-
          tion of NF-kB was detected again.


          Conclusions
          Our findings confirm cold preservation could not prevent oxidative injury during reper-
          fusion in the small bowel tissue. Four cycles of ischemic preconditioning prior to intes-
          tinal cold storage could moderate the injury better preserving the endogenous anti-
          oxidants. IPC before cold preservation was able to moderate oxidative stress inducing
          intracellular signalling pathways, which indicated both cytoplasmic and subsequent
          nuclear NF-kB activation in bowel tissue.




Abstract no.            1
                                                       12
                               13




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                               13
S-NO-HSA has the potential to
optimize preservation strategies
in cardiac transplantation
Semsroth, Severin
Dept. of Cardiac Surgery, University Hospital Innsbruck, Innsbruck, Austria




          Objective
          Depletion of nitric oxide (NO) is asscociatd with ischemia/reperfusion(I/R) injury during
          cardiac transplantation. We tested if NO-substitution with a novel NO-donor, S-Nitroso
          Human Serum Albumin (S-NO-HSA), could bridge NO starvation during early R and
          therefore leads to superior hemodynamic and metabolic outcome during I/R in isolated
          rabbit hearts.


          Methods
          On an isolated red cell perfused Working Heart (WH) model 18 rabbit hearts were per-
          fused according to the following protocol: After assessement of hemodynamic base-
                                ,                                           ,
          line (coronary flow=CF cardiac-output=CO, left atrial pressure=LAP myocardial oxygen
          consumption=MVO2) hearts were randomly assigned into 3 groups. Group 1 received
          S-NO-HSA (0,2 µmol/100ml perfusate, n=8), group 2 received L-arginin (10 mmol, n=8),
          group 3 (control) received albumin (0.2 µmol/100ml perfusate, n=8) only. After 30 min
          of infusion, hearts were arrested with Celsior® (4°C) and stored in the same solution
          for 6h, followed by 60 min of R, before freeze-clamped heart biopsis were taken for the
          determination of high energy phosphates (HEP).


          Results
          During early reperfusion postischemic recovery (%) of CO was significantly higher
          in group 1 (74±6%, p<0.05) compared to group 2 (57±7%) and group 3 (52±4%), CF
          was significantly higher in group 1 (98±7%, p<0.05) compared to group 2 (77±7%) and
          group 3 (74±6%), MVO2 was signifantely reduced in group 1 (68±8%, p<0.05) compared
          to group 3. After 60 min of reperfusion, CO was significantly higher in group 1 (56±5%,
          p<0.05) and group 2 (53±3%, p<0.05) compared to group 3 (36±4%), CF was significantly
          higher in group 1 (67±4%, p<0.05) compared to group 3 (53±5%), LAP was significantly
          reduced in group 1 (118±9%, p<0.05) and group 2 (117±9%, p<0.05) compared to group
          3 (149±10%). There were no significant difference in HEP.


          Conclusion
          In a model of 6 hours of hypothermic cardiac arrest, S-NO-HSA improves hemody-
          namic function, myocardial perfusion and reduces myocardial oxygen consumption via
          preservation of normal endothelium NO synthase function. S-NO-HSA has the potential to
          optimize preservation strategies in cardiac transplantation.




Abstract no.               2
                                                      14
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                               15
Single donor-treatment for the delivery of
carbon monoxide reduces graft immunogenicity
and improves long-term allograft function
P.N.A. Martins1, A. Reutzel-Selke1, A. Jurisch1, K. Atrott1, A. Pascher1, J. Pratschke1, R. Buelow3,
K. Kotsch2, P Neuhaus1, H.D. Volk2, S.G. Tullius1.
             .
1. Department of Surgery, Charité-Virchow Klinikum. Charité-Humboldt University, Berlin
2. Department of Immunology, Charité-Campus-Mitte. Charité-Humboldt University, Berlin
3. Sangstat Medical Corporation, Menlo Park, California-USA.




          Background
          Chronic rejection remains the major obstacle for successful transplantation. Events
          leading to unspecific inflammatory damages occurring prior to organ transplantation
          reduce graft survival. It can be speculated that an increased immunogenicity acceler-
          ates specific immune responses.


          Purpose
          Here, we wanted to test the effects of carbon monoxide (CO) delivery with methylene
          chloride (MC), a CO donor, shortly prior to organ harvesting in a model of chronic rejec-
          tion in the rat.


          Methods
          F-344 rats renal allografts were grafted into Lew recipients (n=6/group). Donor animals
          were either treated with Methylene Chloride (single dose MC 100 mg/Kg p.o.) 4 h prior
          to organ harvesting or remained untreated. Recipients received no MC treatment, short-
          term (10 days) or long-term (180 days) MC treatment (100 mg/Kg/day p.o.). In another
          series, using the same model, grafts were taken 24 h after transplantation for real-time
          PCR analysis. In another set of experiments, using a strong incompatibility model (DA
          to LEWIS), donor-specific MHC class II + dendritic cells (DCs) were determined by haplo-
          type-specific mAb flowcytometry (RTlab, OX62+) one day after kidney transplantation.


          Results
          COHb levels peaked by 3.5 h after MC administration (COHb 5.5%±1.1%), and had
          returned to base-line values by 24 h. Six months post transplantation, proteinuria was
          reduced significantly following MC treatment both, after a long-term application in the
          recipient or after a single treatment in the donor (control: 60±20 mg /24 h vs. MC recipi-
          ent long-term: 11±2 mg /24 h and MC donor short: 15±1 mg /24 h, respectively; p<0.05).
          Morphological alterations characteristic of chronic graft deterioration improved in all
          groups following CO delivery. Improvements were impressive following long-term appli-
          cation in the recipient and even more pronounced following a single donor treatment
          (glomerulosclerosis by 180 days: 20±5% following donor treatment; 30±8% following
          long-term MC application, and 80±10% in controls, p<0.0001 and 0.001, respectively).
          arteriosclerosis (p<0.01), tubular atrophy (p<0.0001,) and fibrosis (p<0.0001). While
          ED1+ M , CD4+ T cells and MHC II expression were significantly (p<0.01) reduced by
          180 days in animals receiving MC long-term; cellular infiltrates and MHC II expression




Abstract no.                 3
                                                         16
were comparable in grafts receiving MC as donor treatment, over a short-term recipient
treatment or in untreated controls. Both, short-term and long-term treated recipient
groups demonstrated functional and structural improved grafts. However, renal func-
tion and morphology were particular better preserved with donor treatment. 24 hrs
after transplantation, grafts from MC treated donors showed a reduced expression of
CD3 (p<0.05, n=4).


Conclusion
CO delivery in the donor was associated with reduced graft immunogenicity, as dem-
onstrated by reduced intragraft infiltration of T-cells, amount of donor-specific DCs and
marked improvement of morphology and long-term graft function.


Keywords
carbon monoxide, donor treatment, chronic rejection, immunogenicity.




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                                         17
Protective effect of p53-pathway blockade
on ischemia-reperfusion injury in a liver
transplantation model in the rat
Frederik Berrevoet, MD, Michael Menger, MD, PhD*
Dept. Of General, Hepato-biliary and Transplantation Surgery, University of Ghent, Ghent, Belgium
and Institute for Clinical and Experimental Research, University of Saarland, Homburg, Germany*




          Introduction
          Liver allografts are exposed to variable periods of cold and warm ischemia during pres-
          ervation and transplantation. By histology, studies concerning cold and warm pres-
          ervation of porcine and human livers demonstrated that apoptotic hepatocytes and
          detachment of sinusoidal endothelial cells may be responsible for early graft failure
          after transplantation.
             In the present study we analysed the kinetics of hepatocellular apoptosis using a
          novel fluorescence microscopic technique in cold-stored livers and evaluated the
          impact of the p53-inhibiting factor Pifithrin- on apoptotic tissue injury.


          Material and methods
          Eight male Sprague Dawley rats (weight range 250–300 g) were treated with Pifithrin-
          (2.2 mg /kg body weight, intraperitoneally) 30 minutes prior to liver harvesting. Seven
          male age-matched, vehicle-treated (DMSO, 100 µl /100 g body weight, ip) animals served
          as controls. After pentobarbital anesthesia (50 mg /kg body weight, ip), laparotomy
          and intra-arterial application of bisbenzimide (H33342, 10 mg /kg body weight) livers
          were harvested via portal venous flushing with 100 ml 4° C HTK solution (100 cm H2O
          pressure) and stored in 4° C HTK solution for 24 hours. Using fluorescence microscopy
          and UV epi-illumination, grafts were analysed for apoptotic cell death, as assessed by
          nuclear chromatin condensation and fragmentation of hepatocytes. After the 24 hours-
          period of cold storage the livers were flushed with 10 ml 4° C Ringer’s lactate solution.
          Liver tissue was processed for routine histology and immunohistochemistry.


          Results
          UV epi-illumination microscopy revealed a progressive increase of the number of apop-
          totic hepatocytes from 1.2%±0.3 to 1.6%±1.2 and 10.8%±2.1 at 1 h, 6 h and 24 h of cold
          storage in the DMSO-treated controls. Importantly, Pifithrin-       significantly reduced
          the number of apoptotic cells by approximately 50% at all time points. Concomitantly
          tissue injury, as assessed by semiquantitive scoring of hepatocellular vacuolisation,
          necrosis and sinusoidal detachment, was found significantly attenuated by p53-inhibi-
          tion using Pifithrin- .


          Conclusion
          p53 is known as one of the factors that regulate cell cycle response after DNA damage.
          Pifithrin- treatment seems to be a promising novel approach to reduce apoptosis of
          both hepatocytes and sinusoidal endothelial cells after cold preservation of the rat liver.
          Further studies have to proof the capability of Pifithrin-   to decrease post-transplant
          graft failure by inhibition of apoptotic cell death.




Abstract no.             4
                                                        18
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                                19
Possible beneficial effect of donor
carbon monoxide poisoning on renal
transplant outcome
Maria Bojakowska
Department of General, Vascular and Transplant Surgery, Warsaw Medical University




         The use of solid organs procured from with carbon monoxide (CO) donors poisoned
         remains controversial. However experimental data suggests that carbon monoxide has
         a potentially beneficial effect on organ transplantation allevieting ischemia/reperfusion
         injury. In our department we procured kidneys from 25 year old man who died due to
         CO poisoning caused by improperly ventilated bathroom with fuel-burning heater. At
         admission patient was in severe condition, unconscious with 3 points on GCS, with inef-
         ficient, sparse, spontaneous breathing and hypotension. Carboxyhemoglobin level was
         48.7%, after 1 hour of ventilation with 100% oxygen this level diminished to 12.1%, and
         after 5 hours to 1.2%. Diuresis was above 250 ml per hour. Serum creatinine levels were
         below 1.0 mg per dl. After 40 hours of observation in ICU he started to present symptoms
         of brain death. After declaration of brain death by medical commission, accordingly to
         national law, his kidneys were procured for transplantation purposes.
            A 46 years old man dialyzed for 20 months by continuous peritoneal dialysis was one
         of the recipients. He had no diuresis. Kidney transplantation procedure was complicated
         by external iliac artery dissection in site of anastomosis. Firstly, we tried to make second
         anastomosis but because of persisted dissection of the artery a short by-pass of external
         iliac artery (7 mm in diameter Dacron prosthesis) was performed. Graft renal artery was
         implanted to the prosthesis. Total second warm ischemia time was significantly prolonged
         and was nearly 100 minutes (45 minutes of first anastomosis, followed by cold perfusion
         of kidney graft and second anastomosis of 16 minutes duration, and third anastomosis
         – 35 minutes long). Total ischemia time was 24 hours and 35 minutes.
            It is known that anastomosis time strongly correlates with delayed graft function. But
         in this case, to our surprise, the graft remained very well perfused after all these ischemias
         and reperfusions and started to produce urine immediately after transplantation. The
         recipient did not need neither haemodialysis, nor diuretics after transplantation. His diu-
         resis, blood creatinine and urea levels after procedure are shown on figure 1. He was
         discharged 10 days after transplantation. Within 1 months after transplantation creatinine
         level was below 1.0 mg per dl, he did not show any signs of rejection. Immunosupression
         was induced by combined cyclosporine A, azathioprine and steroid administration.
            We suppose that this surprising good outcome after transplantation with prolonged
         anastomosis time could be explained by donor CO-poisoning. Pleiotropic effects of CO
         are of recent evidence – CO participates in neuronal transmission, promotes vasorelaxa-
         tion and inhibits proliferation of smooth muscle cells, apoptosis, vascular remodeling,
         xenogeneic rejection, inflammation, platelet aggregation, thrombosis and fibrin deposi-
         tion in the microvasculature, cytokine production, and oxidative stress. These properties
         of CO led to experiments with CO delivered either as a gas or via CO-releasing molecules,
         transition metal carbonyls transporting CO to a target tissues (so called CORMs) to protect
         organ from ischemia/reperfusion injury.




Abstract no.            5
                                                        20
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                               21
To determine the role of cryopreservation
by abdominal cooling in non-heart beating
donor renal transplantation
John F Asher
Liver and Renal Transplant Unit, Freeman Hospital, Newcastle upon Tyne, United Kingdom




         Background
         Renal transplantation is currently the best form of long-term renal replacement therapy,
         in terms of duration1 and quality of life2, and cost-effectiveness3. However, access to trans-
         plantation is limited by the shortage of suitable brain-stem dead cadaveric donors.
               An increasingly-used source of organs is non-heart-beating donors. The renal trans-
         plant unit in Newcastle upon Tyne has extensive experience in non-heart-beating donor
         transplants, and our series now shows long-term graft function and graft survival rates
         almost identical to those encountered in conventional heart-beating transplants4.
               Non-heart beating renal transplants are associated with high rates of delayed graft
         function5, which is likely to be related to acute tubular necrosis and vascular injury
         resulting from the inevitable long primary warm ischaemic times associated with Maas-
         tricht category II donors. Our current practice is to minimise the warm ischaemic dam-
         age prior to the retrieval procedure by in situ cold perfusion via a catheter placed in the
         aorta via the femoral artery. Light and co-workers in Washington DC have reported a
         series of transplants where intraperitoneal cooling was added, leading to a decrease in
         the incidence of delayed graft function6.


         Aims
         The aim of the study is to determine whether the warm ischaemic damage in non-heart
         beating renal allografts can be further reduced by additional cooling, either by intraperi-
         toneal cooling or external cooling by ice applied to the abdominal wall.


         Methods
         This study includes a small model and a human trial.


         Small animal model
         A small animal model using rats is used to test normal intraperitoneal temperature dur-
         ing perfusion, tissue ATP levels and histological changes in tissue sections and per-
         fusate enzymes. Two groups of 24 rats are used, with one group receiving only in situ
         cold perfusion and the other additionally receiving cold peritoneal dialysis fluid at 0º C
         via two cannulae inserted on opposite sides of the abdomen.
               The rats are humanely killed by cervical dislocation, and a rectal temperature probe
         inserted. After a 30 minute wait, an iliac cut down is performed preserving the peritoneum,
         and a 22 gauge cannula inserted into the iliac artery, with another inserted into the iliac
         vein for exsanguination. To match the perfusion protocol used clinically, perfusion is
         started at 30 minutes with Marshall’s organ preservation solution at 4º C and a bolus
         of 10.000 units of streptokinase. The volume of perfusion solution is 80 ml for a 280 g
         rat, which is proportionate to the 20 litres used for a 70 kg human. Core temperature is
         recorded at the commencement of perfusion.




Abstract no.             6
                                                        22
Following a period of 2 hours, a laparotomy is performed and the liver and kidneys removed.
A lobe of the liver and the right kidney is snap-frozen in liquid nitrogen for ATP analy-
sis, and the remainder of the liver and left kidney undergo hypothermic machine per-
fusion for 2 hours to obtain perfusate samples for analysis of biochemical markers of
tissue injury. The liver and kidney are then reperfused with an oxygenated Krebs-Hense-
leit buffered salt solution at 37° C for 1 hour; following reperfusion, a biopsy is taken
from each and snap frozen in liquid nitrogen for ATP analysis, then trypan blue added
to the system in the last 7 minutes to evaluate cell death. The kidney is then be fixed,
embedded in paraffin and stained with for histological analysis; a selection will also
undergo electronmicrography to assess mitochondrial integrity.


Human study
Ethical permission for this has recently been granted, and it will follow earlier proto-
cols of the procedure being randomised to be performed, or not, on potential donors
together with in situ perfusion prior to moving to theatre for laparotomy and retrieval
of kidneys.
   After cannulation for in situ perfusion, a 10 mm laparoscopic port will be inserted
through the umbilicus and the gas insufflation port connected to a 5 litre bag of perito-
neal dialysis fluid at 4º C. A silastic tube will be inserted through the laparoscopic port
into one of the iliac fossae and connected to the first via a heat exchanger and pump
to ensure circulation. The fluid will be circulated until the donor is moved to theatre for
retrieval.
   The opening temperature will be determined with an infrared laser thermometer.
All kidneys will be perfused for viability as per protocol and a time 0 biopsy taken for
ATP levels. The primary endpoints will be kidney discard rate following viability assess-
ment, and incidence and duration of delayed graft function following transplantation.
Tissue ATP concentrations will form a secondary endpoint.


Analysis of results
In the animal study, the endpoints are quantitative measures of ischaemic injury in the
form of levels of biochemical markers of injury, tissue ATP concentrations following
reperfusion, histological evidence of injury and flow characteristics on machine perfusion.
   In the human study, the endpoint will be the incidence and duration of delayed graft
function. The aim will be a reduction in incidence from the current level of 95% to 60%
as described by the Washington and Maastricht groups. Aiming for a power of 80% with
an alpha of 0.02, 24 patients will be used in each group.
(Word count 1020)




                                          23
References
              ,
1. Schnuelle P Lorenz D, et al: Impact of renal transplantation on survival in end-stage
  renal failure: evidence for reduced mortality risk compared with haemodialysis during
  long-term follow-up. Journal of the American Society of Nephrology 9:2135, 1998
2. Parsons DS, Harris DC: A review of quality of life in chronic renal failure. Pharmac-
  oeconomics 12(2)140-60, 1997
3. de Wit GA, Ramsteijn PG, de Charro FT: Economic evaluation of end-stage renal disease
  treatment. Health Policy 44(3):215-32, 1998
4. Gok MA, Buckley PE, Shenton BK, et al: Long-term renal function in kidneys from non-
  heart beating donors: a single-centre experience. Transplantation 74(5):664-9, 2002
                        ,
5. Balupuri S, Buckley P Snowden C, et al: The trouble with kidneys derived from
  the non-heart beating donor: a single centre 10 year experience. Transplantation
  69(5):842-6, 2000
6. Light JA, Sasaki TM, Aquino AO, et al: Excellent long-term graft survival with kid-
  neys from the uncontrolled non-heart beating donor. Transplantation Proceedings
  32(1):186-7, 2000
7. Gok MA, Shenton BK, Peaston R, et al: Improving the quality of kidneys from
  non-heart beating donors using streptokinase: an animal model. Transplantation
  73(2):1869-74, 2002




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                                            24
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                               25
The influence of donor risk factors and
delayed graft function on kidney graft
outcome in the first year
                         .
M. Arnol, A. Kandus, A. F Bren
University Medical Center, Department of Nephrology, Ljubljana, Slovenia




         Outcome of kidney transplantation depends on many variables. This study examined
         the influence of donor risk factors (age, history of hypertension, death from cardiovas-
         cular accident, preharvesting serum creatinine) and post-transplant delayed graft func-
         tion (DGF) on kidney graft outcome in the first year.
             116 adult cadaver kidney recipients, transplanted in our center from October 1999
         through May 2002 were analyzed. All patients received equal immunosuppression with
         basiliximab, cyclosporine, mycophenolate mofetil and steroids. Graft function was
         expressed as serum creatinine at 1, 3 and 12 months and as the slope of 1/creatinine
         beyond 1 month.
             Patient and graft survival at 1 year was 99.1%. During the 1-year period, serum cre-
         atinine steadily improved (119±44, 113±37, and 106±35 M). Mean slope of 1/creatinine
         was 1.4 10 –3 ±4.5 10 –4 M–1 per year. A progressive decline in graft function was noticed
         only in two patients who experienced acute rejection. Accounting for acute rejection,
         multiple linear regression analysis showed that older donor age (each year) (B 0.9, 95%
         CI 0.5 –1.4; P<0.001), preharvesting serum creatinine >100_M (B 15.3, 95% CI 2.4 – 28.1;
         P= 0.02), and post-transplant DGF (B 16.5, 95% CI 4.0 – 28.9; P= 0.001) were independ-
         ently associated with higher values of 1-year serum creatinine. Donor history of hyper-
         tension and cardiovascular cause of death were not found as independent predictors of
         1-year kidney graft function.
             Our results suggest that older donor age, elevated donor serum creatinine and
         post-transplant DGF independently predict less favorable improvement in 1-year kid-
         ney graft function.




Abstract no.           7
                                                      26
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                               27
How to use kidneys from marginal donors:
dual kidney transplantation and calcineurin
inhibitor-free immunosuppression
                                  .
G. Margani L. Furian, N. Baldan, P Rigotti
University of Padova, Italy




          Background
          Due to the continuous shift towards older organ donors, the use of sub-optimal donors
          has become a common clinical procedure. The elderly kidney’s reduced nephron mass
          and functional reserve makes it more susceptible to immunological assault, ischemic
          insult and drug toxicity. To increase the transplanted nephron mass in the case of older
          donors, the transplantation of two kidneys into the same recipient (dual kidney trans-
          plantation, DKT) has been adopted by several centers with satisfactory results. Moreover,
          kidneys from marginal donors may be especially susceptible to calcineurin inhibitor-
          mediated nephrotoxicity.


          Aim
          We evaluated a calcineurin-inhibitor-free protocol for antibody induction, using sirolimus,
          mycophenolate mofetil and steroids, adopted at our center in recipients of a DKT from
          marginal donors.


          Methods
          Between April 2003 and March 2004, 10 DKT from elderly donors (mean age =74±4ys)
          were performed at our center in recipients at low immunological risk (mean age =59±4
          ys). The immunosuppressive protocol included induction with anti-human thymocyte
          globulin (ATG) as follows: 2 mg / kg iv. on day 0, 1.5 mg / kg on day 1, 1.25 mg / kg on day
          2, 1 mg / kg on day 3, and 0.625 mg / kg on days 4 and 5. Sirolimus was administered
          at an initial dose of 8 –10 mg on post-operative day 1, and then 4 – 5 mg to adjust the
          levels to 10 –15 ng / mL. 1 g / day of mycophenolate mofetil was administered by day 5.
          Steroids were tapered from 20 mg to 4 mg / day in 8 weeks.


          Results
          After a mean follow-up of 6±3 months, all patients are alive, 9 /10 have a functioning
          graft (the only graft loss was due to venous thrombosis). One patient experienced
          delayed graft function; acute rejection occurred in 2 /10 patients, and sirolimus was
          substituted with tacrolimus in these 2 cases; in another 2 cases sirolimus was switched
          to cyclosporine due to severe leukopenia and arthralgia. Mean serum creatinine at lat-
          est follow-up was 115±28 mol / L. Urinary fistula was observed in one patient; no other
          complications (e g lymphocele, interstitial pneumonitis, wound complication) occurred,
          but hyperlipidemia was observed in almost all the patients.




Abstract no.             8
                                                        28
Conclusion
Dual kidney transplantation from marginal donors can be performed without any recourse
to calcineurin inhibitors, using a sirolimus-based immunosuppressive regimen in com-
bination with thymoglobulin, mycophenolate mofetil and steroids to obtain excellent
results in terms of graft survival and renal function, without increasing the risk of acute
rejection.




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                                          29
Successful multiorgan transplantation from
a methanol-poisoned brain dead donor
Turcu, Rosana Maria Cristina
Emergency, Ambulance Station, Romtransplant, Bucharest, Romania




         The continuous increasing demand for organs to be transplanted has led to organ
         procurement from acutely poisoned donors. There is a little literature on patients who
         have successfully undergone transplantation that used organs from brain dead poi-
         soned donors. This is a case of successful multiple organ transplantation from a meth-
         anol poisoned brain dead donor.
            A 47-year old man was admitted to the ICU Department 36 hours after the ingestion
         of an unknown amount of presumed ethanol. His relatives described the onset of the
         disease with nausea and vomiting rapidly followed by loss of consciousness. Upon
         admission to the hospital he was comatose (GCS = 3), with fixed dilated pupils, respira-
         tory arrest and bradicardia. Tonic seizures followed. The initial neurologic assessment
         indicated deep coma with severe impairment of brain stem reflexes. The diagnose was
         diffuse metabolic encephalopathy. The cerebral CT scan showed difuse brain swelling
         with severe intracranial hypertension. The routine toxicology screening followed indi-
         cating methanol plasma level of 1.85 g / l.
            The clinical and biological data were consistent with the diagnose of acute metha-
         nol poisoning. Despite sustained intensive care therapy the neurologic condition of the
         patient was continuously deteriorating and the brain stem reflexes (including apnea
         test) disappearing 4 hours later.
            There are still a number of cases of severe methanol poisoning in whom the brain
         may experience irreversible damages, mainly due to the severe metabolic acidosis, whilst
         the function of other organs (liver, kidneys) may recover with proper supportive therapy.
            Organ donation was considered. Ten hours after the admission in the ICU, the trans-
         plant coordination team was informed about the presence of a potential brain dead
         patient with acute methanolic intoxication. The first determination of the brain death
         diagnosis was performed 18 hours after the admission of the patient in the ICU and
         the second 6 hours later. After the brain death diagnosis, the transplant coordinator
         obtained the family consent for organ and tissue donation.
            The examination of the renal function showed good results and the liver and kid-
         neys were offered. The coordinator decided that the offert is conditionned by the
         results of the macroscopic examination of the liver and an extemporaneous biopsy.
         The harvesting of the heart was not indicated due to the long period of metabolic aci-
         dosis. Concerning the tissue transplant, the skin was discfarded due to the diagnosis
         of psoriasis and the family consent was not obtained for cornea, bones and tendons.
         Prior to organ prelevation acid-base disorders were fully corrected and both kidney
         and liver function were preserved. The plasma methanol level at the moment of organ
         removal could not be assessed.
            The harvesting started 4 hours after the family consent and was not longer than 3
         hours and half. A liver biopsy was performed. The examination showed normal fea-
         tures. The liver was transplanted atfer a 4-h cold ischemia time. The recipient was a
         47-year-old woman with a primary biliary cirrhosis. The warm ischemia time was 45
         minutes. At present all the recipients are in good clinical and functional health.




Abstract no.            9
                                                       30
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                               31
High-dose calcineurin inhibitor blocks
the generation of regulatory cells, whereas
low-dose promotes their development
M. Kawai1, H. Kitade1, C. Mathieu1, M. Waer1, J. Pirenne1
Dept. of Abdominal Transplant Surgery, UZ Gasthuisberg, Belgium




          Aims
          Regulatory cells (RC) are increasingly recognized as playing a major role in non-dele-
          tional forms of tolerance and seem to depend upon IL2-signaling. For safety reasons,
          clinical trials of tolerance induction (transfusion of donor-specific hematopoietic cells,
          stem cells etc.) include immunosuppressive (IS) drugs in the early postTx period. It
          is therefore essential that IS protocols do not block RC generation. The effects of cal-
          cineurin inhibitors (dose/timing) on RC generation is not elucidated yet.


          Methods
          We developed a model of RC-based tolerance to cardiac allografts via preTx donor-spe-
          cific blood transfusion (DSBT) in a fully mismatched RA-to-PVG rat combination.


          Results
          Adoptive transfer from tolerant-to-naïve recipients demonstrated that donor-specific
          RC operate in this model. We then exposed DSBT-treated rats to high (50 mg / kg) and
          low (10 mg / kg) dose of Cyclosporine A (CsA) and we found that high-dose CsA given
          at the time of DSBT abrogated the development of RC and caused rejection. High-dose
          CsA at the time of Tx did reduce – but did not abrogate – the tolerogenic effect of DSBT.
          In stark contrast with high-dose, low-dose CsA did not affect DSBT-induced tolerance.
          On the contrary, low-dose CsA alone given postTx (day 0 –11) in DSBT-untreated rats
          induced tolerance and allowed the development of RC that could adoptively transfer
          tolerance in second set naïve recipients. Finally, DSBT given the day of Tx did not pro-
          mote tolerance in this model, unless recipients received a delayed and subtherapeutic
          course of low-dose CsA (day 5–9) (n = 6 in each experiment, p <.01 = significant).


          Conclusions
          High-dose calcineurin inhibitor abrogates RC generation whereas low-dose (alone or
          in synergy with DSBT) promotes their development. These data have important impli-
          cations with regard to a more judicious use of calcineurin inhibitors in the clinic, par-
          ticularly when immunomodulatory strategies based upon RC generation are applied.
          Sustained and profound (but not delayed and mild) inhibition of IL2-signaling prevents
          RC generation.




Abstract no.            10
                                                       32
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                               33
In vivo effects of immunosuppressive drugs
on dendritic cell kinetics in peripheral blood
of patients undergoing renal transplantation
DA Hesselink1, MA Verkade1, P Athanassopoulos1, MGH Betjes1, WCJ Hop2, JNM IJzermans3,
LMB Vaessen1, CC Baan1, W Weimar1
Departments of 1Internal Medicine, 2Biostatistics, and 3Surgery, Erasmus University Medical Center,
Rotterdam, The Netherlands




          Aims
          The effects of immunosuppressive agents on T-cell function have been well character-
          ized but virtually nothing is known about the in vivo effects of these drugs on human
          dendritic cells (DC). We therefore studied the DC number and composition in peripheral
          blood of renal allograft recipients before and after transplantation and in their donors
          before and after kidney donation.


          Methods
          Using flowcytometry, we determined the number of myeloid (m) and plasmacytoid (p)
          DC in peripheral blood of 24 patients with end-stage renal disease (before, 7, 30 and
          90 days after transplantation) and 23 living kidney donors (before, 3 and 90 days after
          donation). mDC were defined as T/B/NK-lymphocyte lineage-, CD34-, HLA-DR+, CD11c+
          and CD123low and pDC as lineage-, CD34-, HLA-DR+, CD11c- and CD123high. Additionally,
          the maturation status of these DC subsets was determined based on the expression
          of CD83 and the chemokine receptor CCR7. All patients were treated with tacrolimus,
          mycophenolate mofetil and corticosteroids.


          Results
          Transplantation resulted in a decrease of total DC numbers from 16.73±6.26 cells/uL
          at baseline to 6.30±5.12 cells/uL on day 7. The total DC count stabilized on day 30 at
          5.73±4.04 cells/uL and increased to 11.05±5.46 cells/uL on day 90, which was still dif-
          ferent from baseline (p<0.001, repeated measurements ANOVA). This pattern of ini-
          tial decrease followed by a partial recovery, was observed in both DC subsets. How-
          ever, the decrease in pDC was more pronounced, resulting in a relative increase of the
          mDC percentage from 73% at day 0 to 85% at week 1. The percentage mDC normalized
          at month 1 (74%) and was slighty skewed towards pDC at month 3 (69%) (p=0.025).
          After transplantation, expression levels of CD83 and CCR7 were comparable between
          time points and showed no evidence for DC activation. In kidney donors, DC counts
          also decreased significantly after surgery to 10.58±5.52 cells/uL on day 3. However, in
          contrast to renal transplant recipients, total DC numbers had completely recovered at
          month 3 (20.33±5.10 cells/uL) and no shift in the mDC to pDC ratio occurred. Again, sur-
          gery did not result in DC activation. The 7 patients treated for (presumed) acute rejec-
          tion (AR) (6 with high-dose corticosteroids, 2 with rATG) had a lower total DC count at
          week 1 compared with non-rejectors (2.55±2.80 vs 7.94±5.09 cells/uL, p=0.016). This dif-
          ference was caused by a significant difference in the mDC count: 2.15±2.41 vs 6.81±4.54




Abstract no.            11
                                                      34
cells/uL (p=0.019) for patients treated and patients not treated for AR, respectively. pDC
numbers at day 7 were comparable between patients who had received anti-AR therapy
and those who did not: 0.40±0.47 vs 1.13±0.96 cells/uL.


Conclusions
Surgical procedures in living kidney donors and their recipients result in a marked
decrease of peripheral blood DC numbers. In donors this effect was transient, whereas
in transplant recipients the DC recovery was far from complete. These findings, taken
together with our observations in patients treated for AR, suggest an important additional
effect of immunosuppression.



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                                          35
PDGF – a potential target molecule for
preventing chronic allograft nephropathy
J. Savikko, E. von Willebrand
Transplantation Laboratory, University of Helsinki, and Helsinki University Central Hospital,
Helsinki, Finland




          Introduction
          Chronic allograft nephropathy still remains the major unsolved problem in clinical kidney
          transplantation. Chronic allograft nephropathy is an irreversible fibrotizing process lead-
          ing eventually to the loss of the graft, currently there is no treatment available for pre-
          venting it. Both immunological and nonimmunological factors contribute to the devel-
          opment of chronic allograft nephropathy.


          Aim
          Here we investigated the role of PDGF (platelet-derived growth factor) in acute and
          chronic renal allograft rejection, also the molecular mechanisms between acute rejec-
          tion and chronic allograft nephropathy were studied.


          Methods
          Kidney transplantation in rat (DA - >WF) was used as a model. Kidney allografts were
          immunosuppressed with CsA (1.5 mg / kg / d sc). One group of allografts was also treated
          with imatinib (10 mg / kg / d po), a selective PDGF receptor tyrosine kinase inhibitor. Syn-
          genic grafts between DA rats were used as controls, no immunosuppression was given
          to these animals. Kidney grafts were harvested 5 and 90 days after transplantation for
          histology and immunohistochemistry. Creatinine and CsA levels were measured once
          a week.


          Results
          PDGF ligands and receptors were induced already at acute renal allograft rejection.
          Acute rejection episodes during the development of chronic allograft nephropathy
          induced even more PDGF and chronic changes. Blocking PDGF receptor signalling with
          imatinib prevented the development of chronic allograft nephropathy. Nearly normal
          kidney function was also seen in imatinib-treated allografts over long-term follow-up.


          Conclusions
          PDGF plays an important role both in acute rejection and chronic allograft nephropathy.
          The early induction of PDGF may start the molecular cascades that link acute rejection
          to subsequent development of chronic allograft nephropathy. Taken together, this study
          helps to understand the molecular mechanisms in chronic allograft nephropathy and
          provides a new candidate drug for preventing it in clinical kidney transplantation.




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                                                         36
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                               37
Does EX VIVO vascular resistance reflect
viability of non-heart-beating livers?
K. Derveaux1, D. Monbaliu1, T. Crabbé1, D. Schein2, J. Brassil2, D. Kravitz2, J. Fevery1,
T. Roskams1, J. Pirenne1.
1
    University Hospitals Leuven, Leuven, Belgium, 2 Organ Recovery Systems, Zaventem, Belgium.




            Aims
            Non-heart-beating (NHB) donor livers could decrease mortality on liver transplant (LTx)
            waiting lists, but their use is limited by a high risk for primary-non-function (PNF) and
            the absence of methods to measure this risk. For NHB kidneys, vascular resistance
            (Vasc Res) during ex vivo Machine Perfusion (MP) is a simple and reliable parameter
            of postTx viability. The purpose of this study was to evaluate ex vivo methods capable
            of predicting organ viability in vivo. The study was designed to determine whether ex
            vivo Vasc Res of livers exposed to various periods of Warm Ischemia (WI) correlates
            with viability postTx.


            Methods
            Porcine livers were recovered after 0’, 45’, and 90’ WI (5 in each group). These WI times
            were selected based upon prior in vivo LTx validation-experiments correlating 0’WI
                                            ,                                         ,
            with normal viability and no PNF 45’WI with altered viability and high PNF and 90’WI
            with complete loss of viability, universal PNF and recipient death. Livers were flushed by
            gravity via the Portal Vein (PV) and the Hepatic Artery (HA) with 10L 4° C Histidine-
            Thryptophane-Ketoglutarate (HTK), and cold stored for 3 hrs. Thereafter, livers were
            perfused with 4° C modified-Belzer solution, using a specifically designed liver MP pro-
            totype (Organ Recovery Systems) including an unlimited flow – pressure controlled (30
            mm HG) and a flow limited (600 ml/min) pressure controlled (7 mm Hg) system for
            HA and PV, respectively. Vasc Res were calculated in vivo after WI during liver flush-
                                                                       .
            out and ex vivo continuously thereafter during 24 hrs of MP Markers of hepatocyte
            damage (AST / LDH) were analysed in the liver effluent during flush-out and in the MP
            perfusate.


            Results
            During flush-out, PV Vasc Res was lower than HA Vasc Res in all WI groups (p<0.0001).
            PV Vasc Res after 90’ WI was higher than after 0’ and 45’ WI (p = 0.003). A similar trend
            was seen for HA Vasc Res (higherVasc Res with longer WI) but this did not reach signifi-
                                       ,
            cance (p = 0.24). During MP PV Vasc Res was low from the start (0 – 0.05 mm Hg. min / ml)
            and remained fairly constant during the 24 hr observation period whereas HA Vasc Res
            was higher at the start but gradually diminished to reach a more constant value after
            4-to-6 hours (0.25 – 0.35 mm Hg. min / ml). This evolution was similar in all WI groups
            and no correlation was observed between HA / PV Vasc Res and WI. AST and LDH in the
            liver effluent at flush-out and in the MP perfusate correlated with WI (p = 0.001).




Abstract no.                13
                                                         38
Conclusions
Contrary to the kidney, Vasc Res during ex vivo MP of NHB livers does not correlate
with the extent of WI damage and does not predict organ viability/function after Tx in
the model studied. Whether modifying the experimental design could unmask this cor-
relation or whether this absence of correlation is inherent in the unique vascular physi-
ology of the liver (double vascular circuit/low resistance sinusoidal network) is under
evaluation. Other ex vivo parameters, among them enzyme release, correlate with WI
damage and viability postTx.



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                                         39
Clinical Experience with AMC
– BioArtificial Liver in Acute Liver Failure
Vincenzo Scuderi
Cardarelli Hospital, Naples, Italy




          Background
          Due to the shortness of donor liver patients with ALF die while on the OLT-waiting list.
          This condition emphasize the requirement of hepatic support system. Bio-artificial liver
          support may improve survival by bridging to liver transplant or own liver regenerate.
          Moreover it has been proposed that ALF patient treated with BAL are in better clinical
          condition, which might positively influence the outcome of OLTx.
              Hereby we report the outcome about safety of phase I clinical trial since 2001 trough
          2003 on 14 patients with Acute Liver Failure (ALF) undergoing to the treatment with Aca-
          demic Medical Centre of Amsterdam Bio Artificial Liver (AMC-BAL) to bridge to OLTx.


          Methods
          The AMC-BAL system consist of an extracorporeal bioreactor which can be filled with
          at least 10x109 viable porcine hepatocytes. The bioreactor, placed in the BAL module, is
          connected in parallel to a plasmapheresis apparatus based on plasma filtration. Patient
          plasma flows through the bioreactor into direct contact with the porcine hepatocytes,
          which are attached to a non-woven polyester matrix.
              Fourteen patients (nine female and five male; aged 21–56 years) were included in
          this trial. Aetiology was: HBV 7; Cripto 3; HAV 1; Wilson 1; LES/AFLP 1; Aman. Ph. 1.
          Twelve patients were included from the Cardarelli Hospital (Naples, Italy) and two
          patients were included from St. Eugenio Hospital (Rome, Italy).
              The AFLP and HAV patients were both pregnant and lost their fetuses during ALF
          prior to BAL therapy. The period between onset of liver disease and BAL therapy was
          from 2 until 11 days.
              All patients had a grade III or IV coma with Glasgow Coma Scale from 5 to 8.
          The patients were placed on the high urgency OLTx waiting list and received BAL treat-
          ment ranging from 4 till 35 hours. Four patients received 2 BAL treatments.


          Results
           All patients showed improved neurological state, improved diuresis and stabiliza-
          tion of hemodynamic during and shortly after the BAL therapy. Also many laboratory
          parameter like bilirubin and ammonia concentrations improved during the treatment.
              The only side effect observed in two patient was short period of hypotension within
          20 minutes after start of treatment. This hypotension was corrected by fluid addition
          and temporary infusion of dopamine. Thirteen patients received OLTx. The median
          waiting time for adequate graft was 30 hours (11–69).
              Nine patients (71.4%) are actually in good clinical condition with a follow-up of 6 to
          32 months. One of this patient did not need OLTx after two treatment because her sta-
          tus improved and it was decided not to transplant.




Abstract no.             14
                                                       40
Four patients (28.6%) died after transplant because PGNF for two patients, irreversible
coma and MOF for the other two.
   At present Porcine Endogenous Retro Viruses (PERV) has been tested in all patients
alive; no PERV RNA and DNA were detected in plasma and peripheral blood monocytes.


Conclusions
This clinical experience shows that treatment with AMC-BAL is a safe and feasible tech-
nique to bridge the waiting time for a liver graft or own liver regeneration. Obviously
we need a controlled clinical study for demonstrate the life supporting capability of the
AMC-BAL.


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                                         41
Comparative study of biochemical and
scintigraphy parameters in liver transplantation
Hassan Kansoul
Department of Transplantation Surgery,
Karolinska University Hospital, Huddinge, Sweden




         Orthopic liver transplantation has dramatically affected the outcome of previously
         fatal end-stage liver diseases. Beside the importance of improved selection of patients,
         advanced surgical techniques, application of new immunosuppressive therapies, cor-
         rect monitoring of the liver function post transplantation has a considerable role in the
         success of the transplantation.
            Scintigraphy is a good method, among other applications, not only to detect hepa-
         tocellular cancer but also to evaluate transplanted liver function. Early postoperative
         monitoring of graft function after liver transplantation relies mainly on bile production
         and biochemical observations. In order to gain more information we have routinely
         performed hepatobiliary scintigraphy during the first week after transplantation. There
         have been reports that the uptake of 99m-Tc-Mebrofenin in the liver might be influ-
         enced by high levels of bilirubin due to their similar carrier mediated transport.
            The hepatic extraction fraction (HEF) is defined by the extraction of 99m-Tc-Mebro-
         fenin by the liver and reflects hepatocyte function (ie: the efficiency of extraction of the
         radiotracer from the blood). It can be assessed visually or quantitatively. Visual inspec-
         tion is usually adequate, but quantification may be required. The HEF is calculated by
         the ratio of the initial hepatocyte uptake divided by the peak vascular uptake:


                 (y intercept exponential fit liver response curve)
         HEF =
                     y max data value liver response curve


         Data from literature indicate an association between hepatocellular disease and low
            .
         HEF However one has to consider that elevated bilirubin levels may inhibit 99m-Tc-
         Mebrofenin transports to the hepatocytes, inducing a false low HEF value. Biliary tract
         obstructions can lead to elevated levels of HEF.
            We here report the result of 61 patients. Majorities were transplanted with cadav-
         eric organs, all immunosuppressed with Tacrolimus and Steroids, or Sandimmune,
         Simulect and Steroids. The age distribution was 7 month to 71 years with a mean value
         of 43 years. The indications for liver transplantation were 17 FAP (Familial Amiloidoses
         with Polineuropathy), PSC (Primary Sclerotic Colangeitis) 5 patients, HCV (Hepatitis
         with Virus C) 10 patients, HBV (Hepatitis with Virus B) 1 patient, HCC (Hepatocellu-
         lar cancer) 7 patients, Biliary atresia 5 patients, Laennec cyrosis (ethanolic cyrossis)
         5 patients and other diseases. Three of which received a liver from living donor in a
         domino procedure. Three children were transplanted with a left lateral segment from a
         related living donor.
            All patients underwent hepatobiliary scintigraphy with 99m-Tc-Mebrofenin during
         the first week after transplantation. HEF was calculated by means of deconvolutional
         analysis and complemented by clearance rate from the blood due to liver uptake, Total
         clearance from the blood due to all possible routes of elimination and Time to the max-




Abstract no.            15
                                                       42
imum uptake in the liver measured from the time-activity curve. Statistical analysis
was performed using linear multiple regression test with confidence 95%.
   Analysing the group of non-FAP patients we found a significant linear correlation
between Clearance rate from the blood due to liver uptake and liver function measured
by all biochemical parameters (peak transaminases, bilirubin, Alkaline Phosphatases,
LDH). However there was no significant linear correlation between hepatic extraction
fraction (HEF) values and biochemical liver function test. These findings indicate that
hepatobiliary scintigraphy with 99m-Tc-Mebrofenin during the first week after trans-
plantation (values of clearance rate from the blood due to liver uptake) is a good early
indicator of liver function. While, we found that HEF has no predictive value in early
liver function evaluation. In the group of FAP patients we found good linear correlation
between clearance rate from the blood due to liver uptake half time and LDH.
   By this analysis we highlighted the importance and practical value of hepatobil-
iary scintigraphy with 99m-Tc-Mebrofenin during the first week after transplantation, in
early evaluation of graft liver function. The findings that in FAP patients no scintigra-
phy parameters were really significantly correlated with biochemical parameters, could
point that early post operative liver scintigraphy is influenced by the liver’s disease.



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                                          43
Living Donor Liver Transplantation for
Recurrent Hepatocellular Carcinoma
Chao Liu, Andrea Frilling, Georgios C. Sotiropoulos, Frank Weber, Silvio Nadalin,
Massimo Malago, Christoph E. Broelsch
Department of General Surgery and Transplantation, University Hospital Essen, Germany




          Long waiting time for a cadaveric graft, associated with a high dropout rate presents
          the main problem in patients with hepatocellular carcinoma (HCC) listed for liver trans-
          plantation (LT). Living donor liver transplantation (LDLT) offers a new possibility to
          avoid this problem. We report on two patients with recurrent HCC treated by LDLT.
             From April 1998 to September 2003, 538 patients, 55 of them with HCC, underwent
          LT at our institution. In 33 patients with HCC, LDLT was performed. Recurrent HCC was
          the indication for LDLT in 2 patients. In a 44-year old non-cirrhotic male patient recurrent
          multicentric HCC was diagnosed 46 months after initial tumor resection. Meticulous diagnos-
          tic work up disclosed no extrahepatic tumor spread. Living donor liver transplantation
          was considered as the only promising therapeutic option. At 19 months after LRLT the
          patient is alive and tumor free. In a 37-year old female patient with negative hepatitis
          serology unilocular, non-resectable recurrent HCC occurred 27 months after partial
          hepatectomy for HCC. Since there was no additional tumor growth, the patient was
          offered LDLT. After uneventful postoperative course, the patient is in good condition and
          without evidence of tumor recurrence at 11th post transplantation month. Both donors
          recovered well and were back to work 3 and 4 weeks after surgery, respectively.
             With the potential donor risk in mind, LDLT represents an appealing option for
          timely treatment of selected patients with recurrent HCC. Its long-term results will con-
          tribute to appropriate selection in this special subgroup of patients and re-evaluation of
          current criteria for LT in HCC.




Abstract no.             16
                                                        44
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                               45
Liver transplantation in patients with
hepatocellular carcinoma: factors implicated
in tumour relapse
B. Gala1, M. Gómez2, F Suárez2, A. Otero2, C. Fernández2, J. Souto2, F Arnal2.
                      .                                               .
1
    Transplant Unit. Hermanos Ameijeiras Hospital. Havana, Cuba.
2
    Liver & Pancreas transplant unit. Juan Canalejo University Hospital. La Coruña, Spain.




            Aims
            This investigation analyses different factors affecting survival after liver transplant for
            hepatocellular (HCC) carcinoma, where macroscopic vascular invasion and tumour
            nodules larger than 5 cm were both identified as risk factors for tumour relapse after
            transplantation.


            Methods
            We revised the cases of patients with hepatocellular carcinoma who underwent liver
            transplantation during the period 1994–2000, and present a detailed analysis of a series
            of variables which may be probably implicated in the appearance of relapse and which
            have an effect on survival.


            Results
            After a mean follow-up of 33 months, the mortality rate was 27.5% and relapse occurred
            in 18.75% of cases. No history of alcohol abuse, the number and size of the nodules,
            the presence of macro and microscopic vascular invasion, and pTNM stage T4 were all
            factors associated with a significantly increased risk of relapse (p<0.05). These factors
            and positive HCV were associated to decreased survival. After a multivariate analysis,
            the size of the nodules and the presence of macroscopic vascular invasion were con-
            sidered the only independent risk factors for tumour relapse and post-transplantation
            relapse and mortality, respectively.


            Conclusions
            Macroscopic vascular invasion and tumour nodules larger than 5 cm are both inde-
            pendent risk factors of tumour relapse after transplantation. Nevertheless, only macro-
            scopic vascular invasion seems to have a significant effect on survival.




Abstract no.              17
                                                         46
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                               47
Recurrent Hepatitis C following liver
trasplantation: Prospective comparative study
between cyclosporine neoral and tacrolimus
                                                                                         .,
Beltrán, B., Berenguer, M., Aguilera, V., Prieto, M., Sempere, L., Rayón, M., San-Juan, F Berenguer, J.
Hepatogastroenterology Service and Liver Unit. Hospital Universitari La Fe. Valencia (Spain)




          Graft cirrhosis is a relatively frequent complication of recurrent hepatitis C, which onset
          has been demonstrated to be related both with the donor age and the inmunosup-
          pression (1). The results published about the role of calcineurin inhibitor drugs on the
          development of graft cirrhosis are controversial.


          Aims
          To establish whether the inmunosuppressor used, cyclosporine neoral (Neo) or tacrolimus
          (Tac), influences the evolution of recurrent hepatitis C.


          Methods
          A prospective randomized study to compare the rate of relevant disease of the graft
          (F3, cirrhosis and colostatic hepatitis) in two groups of liver transplant recipients who
          underwent transplantation between February 2001 and 2002: ( i ) patients receiving Tac
          + prednisone-P- (n = 17) and ( ii ) patients receiving Neo + P (n = 21). Patients co-infected
          with HBV were excluded. Protocol liver biopsies were performed yearly. Patients included
          were those having a protocol liver biopsy and those with colostatic hepatitis independ-
          ently of the follow up.


          Results
          Out of 63 patients undergoing liver transplantation because of HCV cirrhosis during
          the study period, 38 were analized. The remaining 25 were excluded because of asso-
          ciated complications such as: Budd-Chiari n = 2, biliar or arterial complications n = 4;
          non assessable biopsy n = 3; early death n = 16; none because of HCV recurrence. Both
          groups were similar in their basal characteristics: demographics, donor profile, rejection
          episodes, prednisone treatment duration, surgical variables and follow up. At one year
          post-transplantation, the percentage of patients with relevant liver disease was not sta-
          tistically significant between both groups (Neo 29% vs Tac 35%, p = 0.7) (Table 1) with a
          similar cumulative probability during the follow up (p = 0.47).


                             F = 3 (n=6)       F=4 (n=1)       Colestatic Hepatitis (n=4)
           Neo (n=21)               5               1                       0
           Tac (n=17)               1               0                       4
           p                        ns             ns                     0.04

          Patients treated with Tac developed relevant disease sooner than those treated with Neo.




Abstract no.             18
                                                        48
Conclusion
After a short follow up, no differences in the outcome of recurrent hepatitis C were
found between patients using tacrolimus or cyclosporin Neo for inmunosuppression.
(1) Berenguer-M et al Hepatology 36(1): 202 – 2120, 2002


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                                         49
Conversion from Cyclosporin to
Mycophenolate-based Immunosuppression
Prolongs Kidney Graft Survival in Patients
with Chronic Allograft Nephropathy
Alun Williams, Anita Boswell, Linda Evans, Keith Rigg, Magdi Snewta
Nottingham Transplant Unit, Nottingham City Hospital NHS, Nottingham, United Kingdom.




         Introduction
         Management strategies for patients with chronic allograft nephropathy (CAN) remain
         unclear. We have previously reported improved renal function in patients with CAN upon
         conversion from Cyclosporin (CyA) to mycophenolate mofetil (MMF). In this report we
         describe the effect of conversion from CyA to MMF on long-term renal function and
         graft survival.


         Patients and Methods
         The data of 50 patients with biopsy proven CAN converted from CyA to MMF were ana-
         lysed. The estimated time until graft failure (ETGF) based on 1 / serum creatinine was
         calculated using a line-of-best-fit model.


         Results
         Mean time from transplant to biopsy proven CAN was 6.3 years (range 0.4 –16.6). Mean
         follow up time post-conversion was 28 months (range 1– 60). The change in 1 / Cr fol-
         lowing conversion is shown in the graph.
            In 7 patients (14%) ETGF could not be predicted. There were 2 graft failures in this group.
            The ETGF was predictable in 43 (86%) of the patients. Of these patients 18 (42%)
         would have lost their graft by the end of the first year. A further 19 (44%) and 2 (5%)
         would have lost their grafts by the end of the 2nd and 3rd years respectively.
            At the time of analysis 38/43 (88%) were alive with functioning grafts. A total of
                                                .
         36 (84%) had graft function beyond ETGF One graft (2%) was lost within ETGF and 4
         patients died with functioning grafts. Four patients (9%) had MMF withdrawn due to
         intolerable side effects of medication but grafts are still functioning.


         Conclusion
         Our model of predicted graft loss shows continuous improvement in renal function and
         graft survival over time in patients with CAN converted from CyA to MMF.




Abstract no.               19
                                                        50
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                               51
Bkv-associated nephropathy after kidney
transplantation in pediatric recipients
Ghinolfi, Davide
Dept. of General Surgery and Transplantation, S. Martino Hospital,
University of Genoa Genoa, Italy




         Polyoma BK virus-associated nephritis (BKV-N) is a relevant cause of reduced renal
         allograft survival. Currently, patients at risk for BKV nephropathy are identified by the
         presence of cells containing viral inclusion bodies (decoy cells) in the urine, and diag-
         nosis is confirmed by biopsy of allograft tissue. Recently, it has been demonstrated that
         PCR testing for BKV DNA in plasma is a sensitive and specific method for identifying
         viral nephropathy. In a retrospective analysis, we performed PCR assays for BKV DNA
         in plasma and urine samples from 100 pediatric kidney allograft recipients referred to
         our institution in the last 5 years, with the aim of evaluating the incidence and clinical
         relevance of BKV infection in a pediatric cohort. BKV viruria was observed in 26 out
         of the 100 patients (26%), while BKV viremia was concomitantly demonstrated in 5
         patients (5% of the total population). All 5 children with viremia showed renal damage
         consistent with interstitial nephropathy and/or a significant increase of serum creati-
         nine levels. Renal function has stabilised after reduction of immunosuppression in 4
         patients, while the remaining recipient lost the graft due to a histologically confirmed
         interstitial BKV nephritis. Serology analysis performed on graft recipients and donors
         indicate that seronegative recipients receiving the kidney from seropositive donors are
         at higher risk of BKV shedding.
             Data from the literature, and our experience, indicate that treatment of established
         BKV-N has not substantially changed the prognosis, probably due to late intervention.
         However, a pre-emptive therapy approach could represent a more successful strategy.
         In order to identify parameters predictive of BKV-N development to guide pre-emp-
         tive treatment, we analysed by quantitative PCR assay urinary BKV-DNA (qUr-BKDNA)
         in the 24 patients identified with BKV infection/reactivation (positive viruria ± viremia
         in a qualitative assay) in the cross-sectional study. In parallel, BKV-specific cellular
         immune response was evaluated by frequency analysis of IFN -producing T cells in a
         ELISPOT assay. Data obtained in patients with viremia and graft dysfunction (n=5; 3/5
         were biopsied, and showed hystopathology consistent with BKV-N) were compared
         with the results observed in patients with viruria in the absence of viremia, and good
         graft function (n=19). Among the 5 viremia-positive patients, 4 (80%) had qUr-BKDNA
         levels >109, while only 2/19 (11%) viremia-negative patients showed comparable levels
         of qUr-BKDNA. Patients with viremia and/or qUr-BKDNA >109 showed a low or absent
         frequency of BKV-specific T cells, while higher levels were observed in patients with
         qUr-BKDNA <109.
             Our preliminary results indicate negative antibody status of the recipient as the
         most important predictor of active BKV infection, and suggest a correlation between
         presence of viremia and graft dysfunction, urinary BKV load >109 and reduced/absent
         specific immune response. Thus, monitoring of qUr-BKDNA and specific T cells could
         allow to: i) identify patients with active infection at risk of developing BKV-N; ii) under-
         take pre-emptive terapeutic intervention before tissue damage establishment.




Abstract no.            20
                                                       52
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                               53
Prostate cancer in renal transplant recipients:
a single center experience
 .
F Ravera, G. Saredi*, D. Bonucchi, A. Di Felice, E. Rubbiani, A. Savazzi, G. Cappelli, A. Albertazzi
Division of Nephrology Dialysis and Renal Transplantation University Hospital of Modena, Italy
*Department of Urology, University Hospital of Modena, Italy




          The issue of prostate cancer is becoming significant in transplantation as men aged
          over 45 are transplanted more often in the past and prostate cancer incidence is known
          as inscreasing with age.
              There is no consensus regarding the incidence of prostate cancer in renal-transplant
          recipients: in according to literature the common cancers seenable in general popula-
          tion (lung, breast, prostate, colon and invasive uterine cervical carcinomas) showed no
          increase, or even a decrease in this population.
              215 renal-transplant recipients are followed in Modena renal transplant center: 74
          female, 141 male whose 80 are over 45. Our protocol includes screening for serum
          prostate-specific antigen (PSA) two times a year, and for DRE (digital rectal examina-
          tion) if PSA > 2.5 ng / ml. Urologic screening has allowed the identification of 3 cases
          of localized prostate carcinoma: all patients have been treated withperineal prostatec-
          tomy, without pelvic lymphadenectomy.
              CASE 1:64 year-old man, immunosuppressive therapy with cyclosporine and ster-
          oids, 2 years after grafting, after trans uretral resection of prostate (TURP) performed
          for prostatic ipeplasia, incidental finding of prostate cancer, PSA 2.8 ng / ml, negative
          DRE; pT2bNxM0R0, Glason 3+2.
              CASE 2: 52 year-old man, immunosuppressive therapy with tacrolimus and ster-
          oids, 2 years after grafting PSA 10.20 ng / ml, positive DRE; pT2bNxM0R0,Gleason 5+4.
              CASE 3: 66 year-old man, immunosuppressive therapy with cyclosporine, right
          nefrectomy for renal adenocarcinoma (pT1G3NxMx) pretransplantation, PSA 2.68 ng / ml 7
          years after grafting, positive DRE; pT3aNxM0R1, Gleason 4+3.
              In CASE 1 and 2, after 21 and 24 months, PSA is respectly 0.01 and 0.03 ng/ml; in
          CASE 3 PSA is not below the detection limit (PSA 2.2 ng/ml) so the patient undergoes
          adiuvant ormonal therapy and conformational radiation therapy of prostatic cavity.
              The surgical approach to prostatectomy by perineal via allows prevention of pos-
          sible damages to renal and ureteral graft. Furthermore an early prostate cancer diagno-
          sys decreases the need of lymphadenectomy.
              The incidence of prostate cancer in our renal-transplant recipients seems to be
          higher (2.4%) than reported in literature (0.65%–1.9%); this may be due to increased
          age of renal-transplant recipients and to frequency in urological screenings.
              According to guidelines the screening for prostate cancer is suggested yearly.
          According to our experience we suggest a twice a year monitoring of PSA and DRE.




Abstract no.             21
                                                        54
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                               55
Derivation of islet structures and neural cells
from adult rat stem cells
M.Subramaniam, E.Brown*, D.Russell†, AP Payne†, D. J. Deardon, P G. Shiels*
University of Glasgow, Department of Surgery, Western Infirmary Glasgow
* Division of Cancer and Molecular Pathology, Dept. Surgery, WIG; † IBLS Department of Anatomy




          Aims
          To isolate and characterise adult stem cells derived from pancreas.
          To producing islets for the treatment of diabetes.


          Introduction
          Stem cell therapies have been proposed as potential means of treating degenerative
          diseases, such as Parkinson’s and Alzheimers disease, or conditions such as diabetes.
          World wide, 150 million people suffer from diabetes. Available treatments for diabetes,
          or these degenerative conditions are not satisfactory. Despite insulin therapy for diabetics,
          retinopathy, nephropathy, neuropathy are common late complications. We have sought
          to mitigate this situation through the isolation of adult stem cells and the de novo pro-
          duction of islets from such cells, using the rat as a model system.


          Methods
          Minced adult rat pancreatic ducts were cultured in vitro in CMRL for six weeks to gen-
          erate a cell monolayer. Cells derived from this initial monolayer were then further dif-
          ferentiated through growth in DMEM medium and in matrigel, prior to analysis.


          Results
          We successfully derived a population of adult rat islet progenitor cells, which have
          been serially passaged in culture for 11 months without senescing, in keeping with a
          stem cell status. Characterisation of these cells by PCR and immuno-cytochemistry mark-
          ers indicates that they are positive for the expression of the stem cell markers Nestin
          and Oct-4.and negative for PDX-1. Immuno-cytochemistry has also confirmed they are
          positive for the expression of nestin and NCAM.
             Significantly, two distinct populations of cells can be derived from this precursor
          population: islet like cell clusters and neuronal cells. The islet like cell clusters appear
          to be positive for insulin and glucagon transcription, as judged by RT-PCR. They are
          also positive for insulin production on the basis of DTZ staining. Transmission electron
          microscopy has been used to demonstrate that the cell clusters contain a number of
          distinct cell types and has revealed the presence of dense vesicles. These also appear
          to be of distinct types. One cell type possesses vesicles with a dense core and a clear
          halo, typical on insulin containing granules. Others contain no halo, typical o gluca-
          gons and somatostatin vesicles.
             The neuronal cells derived from the same precursor population are positive for neu-
                                                                              ,    ,
          ronal markers nestin, cyc3 and NCAM. They are negative or PDX-1,MBOP GFAP AA3.




Abstract no.             22
                                                        56
Conclusion
We propose that adult pancreas may be a viable source of stem cells from which islets
and neuronal cells may be derived for transplantation. Transplantation studies with
these cells are ongoing.



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                                       57
Analysis of survivors more than 10 years after
heart transplantation in the cyclosporine era
Krzysztof Wróbel
Dept. of Cardiovascular Surgery and Transplantology, John Paul II Hospital,
Jagiellonian University Medical School, Kraków, Poland




          Background
          Heart transplantation (HTX) has a dramatic impact on both life expectancy and quality
          of life in patients with terminal heart failure. Truly long term survival post heart trans-
          plantation has become increasingly frequent over the past two decades.


          Methods
          461 HTX were performed in Department of Cadiovascular Surgery and Transplantology
          between 1988 – 2004. 43 (43.4%) of 99 patients who received transplants between 1988–1994
          have survived more than 10 years after transplantation. Up to now, 39 patients (39.4%)
          are followed up. The patients and their medical records (eg. cardiac catheter studies,
          echocadiography, ergospirometry ) were examined to determine their somatic and cardiac
          status. Psychologic, social, and occupational status and quality of life data were assessed
          by self-rating questionaires.


          Results
          70% of this group of 99 patients survived 1 year, 50% of patients 7 years, 43.4% 10
          years and more. Male patients accounted for 81.3%. Mean follow up was 11.44 years.
          Age of recipients at HTX was 43.82 years, age of donors at HTX 30.95 years. We found
          generally excellent functional status in these patients, but a high incidence of hyperte-
          sion, renal dysfunction, and graft CAD as well as malignancy.


          Conclusion
          The patients surviving more than 10 years after HTX are mostly in good physical status,
          the quality of life is comparable to the general population, and only a few of them have
          significantly limited in their life style. They do show the substantial chronic side effects
          of long term immunosuppression, remaining treatment dependent for a lifetime.




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                               59
contact details
participants
 Arnol, Miha                        Bojakowska, Maria                        Ghinolfi, Davide
 Dept. of Nephrology,               Dept. of General, Vascular and           Dept. of General Surgery and Trans-
 University Medical Center          Transplant Surgery, The Medical          plantation, S. Martino Hospital,
 Zaloska cesta 7, 1000 Ljubljana,   University of Warsaw Banacha 1a,         University of Genoa
 Slovenia                           PL-02-097 Warsaw, Poland                 Largo Rosanna Benzi 10,
 Tel: +386 1 5223121                Tel: +48 22 8230291                      I-16132 Genoa,
 Fax: +386 1 5222282                Fax: +48 22 6595455                      Italy
 miha.arnol@mf.uni-lj.si            mbojak@wp.pl                             Tel: +39 010 5553108
                                                                                   +39 328 2185278
 Asher, John F.                     Derveaux, Katelijne                      Fax: +39 0185 320053
 Liver and Renal Transplant Unit,   Dept. of Abdominal Transplant            dghinolfi@yahoo.it
 Freeman Hospital                   Surgery,
 High Heaton,                       UZ Gasthuisberg                          Hesselink, Dennis
 NE7 7DN Newcastle upon Tyne,       Herestraat 49,                           Dept. of Internal Medicine, Renal
 United Kingdom                     B-3000 Leuven,                           Transplant Unit, Room Ee 563 A,
 Tel: +44 191 233 6161              Belgium                                  Erasmus Medical Center
 john.asher@btinternet.com          Tel: +32 16 348727                       Dr. Molewaterplein 50,
                                    Fax: +32 16 348743                       NL-3015 GE Rotterdam,
 Beltran, Belen                     katelijne.derveaux@med.kuleuven.ac.be    The Netherlands
 Dept. of Gastroenterology and                                               Tel: + 31 10 4635421, pager 5676
 Hepatology,                        Ferencz, Andrea                          Fax: + 31 10 4089443
 Hospital Universitario La Fe       Dept. of Surgical Research and           d.a.hesselink@erasmusmc.nl
 Avda. Campanar 21,                 Techniques, University of Pécs,
 E-46009 Valencia,                  Faculty of Medicine                      Kansoul, Hassan A.
 Spain                              Kodály Zoltan Street 20,                 Div. of Transplantation Surgery,
 Tel: +34 96 3862700/1973107        H-7624 Pécs,                             Center for Surgical Sciences,
 Fax: +34 96 1973118                Hungary                                  Karolinska Institute
 belendevalen@yahoo.com             Tel: +36 72 535820                       B 56, S-141 86 Stockholm,
 belenniclos@hotmail.com            Fax: +36 72 535821                       Sweden
 beltran_belen@yahoo.es             andrea.ferencz@aok.pte.hu                Tel: +46 8 58582568
                                    andrea.ferencz@webmail.hu                Fax: +46 8 7743191
 Berrevoet, Frederik Christiaan                                              Hassan.Kansoul@cfss.ki.se
 Dept. of General, Hepatobiliary    Gala López, Boris L.
 and Transplantation Surgery,       Liver & Pancreas Transplantation Unit,   Kawai, Masaru
 University Hospital Ghent          Juan Canalejo University Hospital        Dept. of Abdominal Transplant
 De Pintelaan 185,                  Xubias de Arriba 84,                     Surgery, UZ Gasthuisberg
 B-9000 Ghent,                      E-15006 A Coruña,                        Herestraat 49,
 Belgium                            Spain                                    B-3000 Leuven, Belgium
 Tel: +32 9 2404892                 Tel: +34 98 1178170                      Tel: +32 16 348727
 Fax: +32 9 2403891                      +34 6581 38125                      Fax: +32 16 348743
 fberrevoet@hotmail.com             Fax: +34 98 1178016                      kawai4@hotmail.com
                                    bgala@infomed.sld.cu                     Sur080@poh.osaka-med.ac.jp
                                    gala32486@yahoo.com




                                                         60
Liu, Chao                             Savikko, Johanna                   Turcu, Rosana Maria Cristina
Dept. of General Surgery              Transplantation Laboratory,        Emergency, Ambulance Station,
and Transplantation,                  Haartman Institute,                Romtransplant
University Hospital Essen             University of Helsinki             Mihai Eminescu 224-226,
Hufelandstr. 55,                      Haartmaninkatu 3,                  Sector 2, Bucharest,
D-45122 Essen,                        FIN-00014 Helsinki,                Romania
Germany                               Finland                            Tel: +40 21 210 5205/250 8946
Tel: +49 201 7231104                  Tel: +358 9 19126592/96                 +40 724 392093
Fax: +49 201 7235946                       +358 50 5880434               Fax: +40 21 210 2676
liuchao@postmaster.co.uk              Fax: +358 9 2411227                rosana_turcu@hotmail.com
mdliuchao@hotmail.com                 johanna.savikko@helsinki.fi        cris2369tpm@yahoo.com

Margani, Giuseppe                     Scuderi, Vincenzo                  Williams, Alun Rhys
Dept. of Surgical and Medical         Dept. of Hepato-Bilio-Pancreatic   Children & Young People’s
Sciences, Ospedale Giustinianeo       Surgery and Liver Transplant       Nephrology Unit,
Clinica Chirurgica Generale 3^,       Center A. Cardarelli Hospital      Nottingham City Hospital NHS
Via Giustiniani 2, I-35128 Padova,    Via A.Cardarelli, 9,               Hucknall Road,
Italy                                 I-80131 Naples, Italy              NG5 1PB Nottingham,
Tel: +39 049 8213173/8212256/         Tel: +39 081 7472372               United Kingdom
      8211322                              +39 339 4819754               Tel: +44 115 9691169
Fax: +39 049 8213152                  Fax: +39 081 7472330               Fax: +44 115 9627678
giuseppe.margani@unipd.it             enzo_scuderi@inwind.it             alun@arwilliams.fsnet.co.uk
traprepa@unipd.it                     dott_scuderi@inwind.it             alunrwilliams@yahoo.co.uk

Martins, Paulo Ney A.                 Semsroth, Severin                  Wróbel, Krzysztof
Dept. of Surgery and Transplanta-     Dept. of Cardiac Surgery           Dept. of Cardiovascular
tion, Charité – Virchowklinikum       University Hospital Innsbruck      Surgery and Transplantology,
Augustenburger Platz 1,               Anichstrasse 35,                   John Paul II Hospital
D-13353 Berlin,                       A-6020 Innsbruck,                  Jagiellonian University Medical
Germany                               Austria                            School, ul. Pradnicka 80,
Tel: +49 30 40003070                  Tel: +43 512 50480812              PL-31-202 Kraków,
paulo.martins@charite.de              Fax: +43 504 22528                 Poland
drpauloney@yahoo.com                  severin.semsroth@uibk.ac.at        Tel: +48 12 614 30 75
                                                                         Fax: +48 12 423 39 00
Ravera, Federica                      Subramaniam, Murali                kardiochir@szpitaljp2.krakow.pl
Division of Nephrology, Dialysis      Dept. of General Surgery,          kwrobel@mp.pl
and Transplantation, Policlinico di   Stobhill Hospital                  krzysztof_wrobel@hotmail.com
Modena                                133 Balornock Road,
Via del Pozzo 71,                     Glasgow G21 3UW,
I-41100 Modena,                       United Kingdom
Italy                                 Tel: +44 141 2112000, bleep 3035
Tel: +39 059 4222473/4222293          murali.subramaniam@lycos.com
      +39 328 7678662
      +39 0171 681869
Fax: +39 059 4222167
federicaravera@yahoo.it




                                                   61
contact details
faculty members
John Dark                           Jan Lerut                             Johann Pratschke
Freeman Hospital                    Dept Abdominal transplantation,       Department of Surgery
Dept of Cardiopulmonary             hepatobiliary and endocrine surgery   Division of Transplantation
Transplantation                     University Hospital St Lus            Augustenburger Platz 1
Cardiothoracic Center               Av Hippocrates 10                     D-13353 Berlin
Freeman Road, Room 115              1200 Brussels                         Germany
Newcastle upon Tyne NE7 7RN         Belgium                               Tel: +49 30 450 65 23 47
United Kingdom                      Tel: +32 27 64 14 12/53 06            Fax: +49 30 450 52 900
Tel: +44 191 284 3111               Fax: +32 27 64 90 39                  johann.pratscke@charite.de
Fax: +44 191 223 1152               lerut@chir.ucl.ac.be
j.h.dark@ncl.ac.uk
                                                                          Henrik Thorlacius
                                    Nils H Persson                        Dept of Nephrology
Henrik Ekberg                       Dept of Nephrology                    and Transplantation
Dept of Nephrology                  and Transplantation                   University Hospital
and Transplantation                 University Hospital                   SE-205 02 Malmö
University Hospital                 SE-205 02 Malmö                       Sweden
SE-205 02 Malmö                     Sweden                                Tel: +46 40 33 10 00
Sweden                              Tel: +46 40 33 10 00                  Fax: +46 40 33 62 11
Tel: +46 40 33 10 00                Fax: +46 40 33 62 11                  henrikthorlacius@hotmail.com
Fax: +46 40 33 62 11                nils.h.persson@skane.se
henrik.ekberg@kir.mas.lu.se
                                                                          David C Wheeler
                                    Robert Porte                          Centre for Nephrology
Olle Korsgren                       University Medical Center Groningen   Royal Free and University College
Department of Clinical Immunology   Department of Surgery                 Medical School
Akademiska University Hospital      Section of Hepatobiliary Surgery      Rowland Hill Street
SE-751 85 Uppsala                   and Liver Transplantation             London NW3 2PF
Sweden                              Groningen University Medical Center   United Kingdom
Tel: +46 18 611 41 87                .O.
                                    P Box 30.001                          Tel: +44 20 7830 2930
Fax: +46 18 611 02 22               9700 RB Groningen                     Fax: +44 20 7830 2125
olle.korsgren@klinimm.uu.se         The Netherlands                       d.wheeler@rfc.ucl.ac.uk
                                    Tel: +31-50-3612896
Dirk Kuypers                        Fax: +31-50-3614873
Dept Nephrology and                 r.j.porte@chir.azg.nl
Renal Transplantation
University Hospital Gasthuisberg
University of Leuven, Belgium
Herestraat 49
B-3000 Leuven
Belgium
Tel: +32 16 34 45 80
Fax: +32 16 34 45 99
dirk.kuypers@uz.kuleuven.ac.be




                                                        62
                                              FS 04059




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