Abstract: The Task Force on Organ Transplantation (DHHS,1986) addressed the issue of increasing organ donation. The Report of the Task Force recommended that "educational efforts aimed at increasing organ donation among minority populations be developed and implemented, so that the donor population will more closely reflect the ethnicity of potential transplant recipients, in order to gain the advantage of improved donor and recipient immunologic matching (DHHS,1986). Donor rates for minorities has increased as follows: 16% in 1988 to 23% in 1995 among cadaveric donors and 24% in 1988 to 28% in 1995 among living donors. The improvement in donor rates among minorities may positively affect the transplantation success rate experienced by organ recipients of the same race. Strategies must be implemented that will increase the effectiveness and frequency of communication between minority patients and the medical community. An increase in the effectiveness of communication between potential minority donor families and the health care community will contribute to the process of Consciousness Raising as discussed by Prochasksa. The result of increased awareness of the organ donation and transplantation process may have a favorable impact on organ donation. The media has, through public service announcements, paid advertising and entertainment programming, attempted to promote discussion of organ donation in the community and within families. Johnson et al. discussed Mexican-American and Anglo-American Attitudes Toward Organ Donation. The primary impediment contributing to the disparity of consent rates between Mexican-American and Anglo-American population occurs with regard to the donation of organs of relatives. Johnson stated that this impediment to organ donation can be effectively addressed by promoting family discussion. Communication within families will inform surviving next of kin of the desire of the deceased to be an organ donor and hence improve the likelihood of the donor family consenting to organ donation (Johnson et al., 1988). The promotion of communication within families must continue to be a goal of the transplant community regardless of race/ethnicity. Despite the efforts of the government, the transplant community, the media and the corporate sector to address the critical shortage of donors in the United States, the reality is that no community has the supply of donor organs suitable to meet the need.

Abstract: Objectives The European Commission funded EUROTOLD Project sought to examine the legal and ethical implications of living donor organ transplantation within Europe, facilitated by a multi-centre study acquiring data on practices, laws, policies, attitudes and decision-making processes. Methods Methods involving primary sources included interviews with clinical staff and past and present organ donors and recipients, and questionnaire surveys of transplant centres, individual clinical staff and legal experts. These strategies were supplemented by an examination of secondary sources such as official reports, transplantation literature, etc. Results The surveys generated substantial new evidence relating to transplant centre policies and practices, and the attitudes of physicians toward living donation generally and the use of certain specific classes of donor. The latter was facilitated by the use of case scenarios, providing a window upon factors influencing judgments in this sphere. The interview data confirmed earlier findings about donor decision-making. Conclusions Responding centres and clinicians displayed a fairly liberal attitude toward living donation but substantial diversity nonetheless exists with regard to living donor transplant volumes between centres and surrounding acceptable waiting times for transplant. Further research is required to identify precisely the reasons underpinning such disparity.

Abstract: The use of xenografts could relieve the chronically inadequate supply of human organs for transplantation, but doubts have been expressed about the general acceptability of transplanting animal organs into human. Some researchers and clinicians have chosen to ignore negative attitudes towards clinical xenotransplantation, assuming that people will automatically embrace this new technology when it becomes available. A review of eight studies of attitudes to xenotransplantation did not reveal overwhelming support for it. Particularly negative views were expressed by acute care nurses. Primates have been the donors of choice in clinical xenotransplantation to date, but their continued use is a highly contentious option; the preferred animal donor is now clearly the pig. Animal farming for xenotransplantation is generally regarded as acceptable since animals provide food for man and are an accepted source of items for human use such as heart valves and insulin. Open debate about xenotransplantation must now take place, and present attitudes may change as a result of this. However, it remains to be seen whether xenografts will be widely accepted and used, and the extent to which the chronic shortage of organs for transplantation will thereby be alleviated.

Abstract: Human organ transplantation is practiced in local cultural worlds that shape beliefs about appropriate conduct for its development and application. The psychological response of individuals to the transplant experience mediate and condition its life-changing force in the context of family and community. In this paper, three cases are examined to illustrate the impact of cultural and psychological influences on human organ replacement therapies. First, we explore brain death and its implications for the definition of death and the procurement of organs. A case example from Japan provides the framework for addressing the cultural foundations that contribute to perceptions of personhood and the treatment of the body. Second, we examine marketing incentives for organ donation using a case from India where, until recently, explicit forms of financial incentives have played a role in the development of renal transplantation involving non-related living donors. Third, we focus on the psychological remifications of organ transplantation using a case that demonstrates the profound experience of being the recipient of the "gift of life". Resolution of scientific and ethical challenges in the field of organ transplantation must consider the complex and significant impact of cultural and psychological factors on organ replacement therapies.

Abstract: OBJECTIVES AND METHODS 219 heart transplant recipients with survival over 3 months were retro- and prospectively analysed for the incidence of primary neoplasms. Patients received immunosuppressive drugs (cyclosporine A, azathioprine, steroids) with a 4-5 days induction course of Rabbit Anti-Thymocyte Immunoglobulin (RATG) or monoclonal antibodies induction /OKT3/ in some cases. Anti-rejection treatment consisted of pulse doses of methyloprednisolon or RATG. RESULTS 9 cases of malignancy (4.1%) with one case of pre-malignant liver condition (dysplasia gigantocellulare, 0.45%) were found (8M; 1F; age: 45-67 y.o., x57.7). Symptoms of neoplasms occurred 7-79 months (x31.4) postoperatively. Skin carcinomas: planoepitheliale, spinocellulare, soft tissue neoplasms/mesenchymal sarcoma, larynx Ca planoepitheliale, lung: adenocarcinoma and Ca microcellulare, kidney Ca clarocellulare and post transplant non-Hodgkin lymphoma were diagnosed. Chemo- and radiotherapy, surgery and reduction of immunosuppression did not change the outcome of malignancy in 6 pts.; (regression-1 pt was., remission-2 pts). Patients died 7-86 months after Htx (x41), 4-25 mos. (x12.5) after suffering from first symptoms and 0-10 months (x4.9) after pathology-based diagnosis of neoplasm. CONCLUSIONS Heart transplant recipients have an increased risk of carcinogenesis. The incidence of malignancies in the studied group is similar or even lower than in other reports.

Abstract: Gingival hyperplasia is a well recognised complication of cyclosporin A therapy. Although its pathogenesis is still debated in several recent reports a second generation macrolide antibiotic-azithromycin induced partial or even complete regression of hyperplasia. We present a patient after kidney transplantation treated with cyclosporin who developed very advanced gigival overgrowth (stage 3+). The patient received a 3-day treatment with azithromycin which was repeated after 3 months. The first course of the drug caused a partial regression of gingival hyperplasia during following months but the repeated treatment did not provide a further regression of the changes.

Abstract: The fluorescence dye Hoechst 33342 (Ho342) is employed for isolating early haematopoietic cells and the aim of this study was to evaluate the in vivo and in vitro toxicity of this compound. First, by employing a murine model we studied the influence of this dye on the morphology of the different organs of animals that have been injected intravenously with increasing doses of Ho342. Accordingly, we found that Ho342 at relatively low doses (0,3 M) caused morphological changes in the spleen and lungs and at higher doses (1,5 & 6 M) damaged also the liver. In contrast, kidneys appear to be relatively resistant to this dye. Next, since Ho342 is employed for isolating early haematopoietic cells by FACS, we have been looking for potential toxicity of this dye against normal human haematopoietic progenitors. Accordingly, CD34+ cells isolated from cord blood (CB) samples were exposed to increasing doses of Ho342 (0-50 microM). We found that the low concentration of Ho342 (10 microM) recommended for isolating HSC significantly inhibited the clonogenecity of human erythroid progenitors (BFU-E). The higher doses of Ho342 have also been toxic against normal human myeloid progenitors (CFU-GM). This study shows that Ho342 could potentially damage human cells. This fact should be considered whenever Ho342 has to be employed for isolating living cells.

Abstract: Fibrinolytic disturbances are thought to play an important role in processes leading to deterioration of renal allograft function. We investigated the effect of CsA therapy on the regulation of fibrinolysis in kidney graft recipients by measuring plasma concentration and activity of plasminogen activators (tPA, uPA) and their inhibitors (PAI-1, 2). We found an increase in tPA activity and in PAI-1 concentration as well as a decrease in PAI-1 activity in renal allograft recipients as compared to healthy controls, but did not confirm a correlation between these observations and CsA administration. tPA and PAI-2 concentrations as well as uPA activity did not significantly differ between the studied groups. We showed a significant decrease in uPA plasma concentration in patients treated with azathioprine. The significance of this finding is unknown.

Abstract: We have noticed that bone marrow transplanted in a vascularized limb graft providing a continuous supply of donor BMC may prolong the survival time of skin graft from the same donor. The question arises whether the raised microchimerism plays a role in the prolonged survival of skin allograft. The aim of the study was to follow the development of microchimerism after allogeneic vascularized bone marrow transplantation (VBMTx) concomitantly with the rejection processes of transplanted skin. The BN rats served as donors and LEW rats as recipients of VBMTx and free skin flap allograft. Hind limb was transplanted followed by a full-thickness skin graft on the dorsum. Cellular microchimerism was investigated in recipients of VBMTx and skin grafts in blood, spleen, mesenteric lymph node and bone marrow with monoclonal antibody OX27 directed against MHC class I polymorthic RTI on BN cells and quantitatively analysed in FACStar. In VBMTx group free skin flap survived 70 days after weaning of CsA. Intravenous infusion of BMC in suspension equivalent to that grafted in hind limb did not prolong skin graft survival after cessation of CsA therapy. Donor-derived cells could be detected in VBMTx recipients as long 70 days after wearing of CsA but not in recipients of i.v. suspension BMC grafting.

Abstract: Kaposi's sarcoma is an unusual neoplasm that is seen with increased incidence in transplant recipients. Its occurrence in this group as a tumor is noteworthy in that it may regress spontaneously if immunosuppression is reduced or discontinued. Thus, its response in this setting raises the question of whether it might be best classified as a reversible hyperplasia rather than a true malignancy. In this work we will review recent advances in our understanding of Kaposi's sarcoma, while retaining an historical perspective.

Abstract: Cyclosporin A (CsA) changes the distribution of the circulating pool of lymphocytes and decreases their traffic to organ allograft. The mechanism of this process is complex and includes, among others, inhibition of induction of nuclear factor of activated T cells and suppression of GM CSF and E-selectin expression. We studied the expression adhesion molecules CD11a, CD18, CD44, CD54 and CD62L on the thoracic duct lymphocytes of rats treated with CsA. The 7-day administration of CsA evidently decreased the expression of CD62L but did not affect the other adhesion molecules. Lower concentration of CD62L molecules on the surface of circulating lymphocytes may influence their migration to allograft and distribution in host lymphoid tissues.

Abstract: Increase of number of organs for transplantation depends on the positive attitude of the general public toward cadaveric organ donation and transplantation. This attitude is shaped as the result of education performed by mass media and as a result of individual experience of various people with the health-care service. Whenever a decision on post-mortem donation of a deceased is unknown, there are his close relations who might express his will. The aim of this article is to draw attention to several problems of psychological nature, which refer to proceeding with relatives of organ donors. A course and result of talks with relatives of a potential organ donor depend on a number of factors, which are pointed out below. Within the frames of this interaction, basic rules of effective communication are presented, including message on death of a close person in case of brain stem death diagnosis. Furthermore, examples are presented of the most frequent errors of those who talk with relatives, scope of appearing difficulties and several methods to avoid and/or overcome occurring problems.

Abstract: OBJECTIVES Modification of the total body irradiation technique before bone marrow transplantation has been implemented to improve uniformity of the dose inside the irradiated body and to obtain appropriate reduction in doses in critical organs. METHODS The total dose of 12.6 Gy was delivered with the use of a combination of the following fields: lateral, anterior-posterior, field to mediastinum, an electron field to the thorax wall and field to coxa. The doses were calculated for ten transverse body sections. Methods of dose reductions in critical regions by splitting different fields were proposed. RESULTS The resulting dose deviations inside the body with the combination of all the implemented fields varied from 12.5 Gy to 13.6 Gy along the midline and from 12.4 Gy to 13.7 Gy outside the midline. Doses to the lungs were reduced to 8.6 Gy divided Gy 9.4 Gy, whereas those in the thorax wall and mediastinum were increased to 12.7 Gy divided by 13.5 Gy respectively. Doses to lenses were 12.9 Gy divided by 13.1 Gy.

Abstract: Immune events developing in the bed of skin allograft and draining lymphatics and lymph nodes are probably a copy of what happens in skin after invasion by bacteria, viruses or fungi. The mechanism of local immune response developed in skin during the evolution and is highly conserved and efficient in elimination of foreign antigens. This is why the take of a skin allograft is so difficult and immunosuppressive measures applied after allogeneic skin transplantation remain so ineffective. Authors present their results of studies on the human skin immune humoral and cellular factors, underlining their specificities and differences compared with blood. They also analyze the role of non-immunological factors, such as tissue fluid formation and lymph flow in transportation of alloantigen to the lymph nodes. The migratory properties of immune cells are an indispensable factor in transfer of alloantigen to the lymph nodes. Understanding of the evolutionarily developed immune events in skin may allow to analyze the process of skin allograft rejection and can give hints for more effective immunosuppressive policy.

Abstract: Insulin-dependent diabetes mellitus is an autoimmune disease caused by the selective destruction of islet beta cells. Allo or xeno transplantation of islet cells may establish a novel promising method of IDDM therapy. Understanding how lymphocytes recognize beta cell antigens is essential for the elucidation of the pathogenesis of islet dysfunction. Leukocyte adhesion to the target cells (endothelium, islets) via adhesion molecule pathways plays an important role in auto and allo/xeno antigen recognition and effector cytodestruction of target cells. However, the expression of these molecules on the endothelium and islet cells during the rejection process still remains unclear. There are some publications describing possible roles of these antigens in the response to the graft. The expression of some of adhesion molecules may contribute to a new method for the diagnosis of graft rejection and its therapy when adhesion blocking substances are used for the treatment.

Abstract: Mitochondrial energetic and oxidative dysfunctions caused by free radical production trigger release of proinflammatory cytokines involved in organ rejection. The aim of this study was to investigate the role of a fluoroquinolone drug, pefloxacin (PFX) and those of various cold preservation solutions on pancreatic beta cell viability. Our data clearly demonstrate that islet cell viability, as determined by glucose-stimulated insulin secretion, is directly correlated with reduced expression of microsomal cytochrome P-450IIIA. Moreover, IL-2, a known mediator of apoptosis was found to be downregulated, whereas TNF-alpha had been upregulated for the first 18 hours after pefloxacin administration. These results demonstrate that pefloxacin downregulates the expression of cytochrome P-450IIIA isozyme and regulates the production of TNF-alpha and IL-2. Thus, we postulate that the presence of pefloxacin in the pancreatic islet cells before organ preservation facilitates increased cell viability.

Abstract: In the previous studies we showed that vascularized bone marrow graft (VBMtx) in transplanted rat hind limb brings about complete repopulation of syngeneic recipient BM cavities and lymphoid organs within 10 days. Transplantation of an equivalent number of bone marrow cells (BMC) in suspension did not produce repopulation until day 30. In this study we present data on transplantation of allogeneic VBM and compare them with those obtained in a syngeneic combination. In the LEW or BN to LEW combination BM cells were labelled with 51Cr, injected i.v., 24 h later the hind limb was amputated and transplanted to a LEW rat. BM cells emigrated from the transplanted limb to the recipient and distributed in BM cavities and lymphoid tissues. In the LEW to LEW combination the level of radioactivity in recipient tibia was after 24 h 0.85, in spleen 2.43, in mesenteric lymph node 0.52%/g of tissue, whereas in the BN to LEW model it was 0.11, 1.83 and 0.15%, respectively. The calculated numbers of BM cells which populated recipient tissues were 8-10-times lower in the allogeneic compared with syngeneic combination. This was probably due to the nonspecific elimination of some subsets of BM cells (allogeneic BMC cytotoxicity). Administration of anti-asialo GMI antiserum to the recipient abrogated the cytotoxic effect. Taken together, major differences in kinetics of seeding and repopulation of BMC from VBMTx were found. Elimination of recipient NK cells with AAGMI antiserum attenuated the nonspecific cytotoxic effect. This protocol allows protection of the grafted BMC and increases the efficacy of the transplant.

Abstract: Transplantation of whole pancreas or pancreatic islets remains a promising approach to treatment of diabetes mellitus. Since there is no efficient method presently known for in vivo detection of pancreatic islet rejection, we have utilized dithizone [DTZ] to monitor the survival of transplanted islet allografts following the induction of tolerance by a new strategy of deliberate introduction of donor antigens into the adult thymus. In this study, we examined the morphology of islet allografts in vivo and in vitro following pretreatment with intrathymic (IT) inoculation of 2 mg soluble Ag obtained from 3M KCl extracts of resting T-cells with or without ALS immunosuppression in the WF-to-Lewis combination. Fresh isolated rat islets stained pink 3-5 minutes following exposure to medium containing 0.12 mM DTZ solution in DMSO. Intravenous (i.v.) injection of DTZ solution into unmodified recipients of islet allografts that had rejected their grafts showed massive degranulation of islets which did not stain pink with DTZ. This was confirmed by microscopic finding of fibrosis and lymphocytic infiltration. In contrast, i.v. injection of DTZ solution into long-term recipients of islet allografts at 50, 100, and 150 days after transplantation showed viable islet cells which stained crimson red with DTZ and the findings were confirmed with microscopic sections. This study demonstrates that DTZ is an effective means of in vivo and in vitro identification of transplanted pancreatic islets and suggests that this strategy may have potential clinical application in the diagnosis of the pancreatic islet rejection.

Abstract: Organ allografts survive in hosts treated with immunosuppressive drugs. The question arises as to whether cells isolated from organ or tissue of an allogeneic donor and transplanted to a genetically disparate recipient can also benefit from the immunosuppressive regimen. We reported previously that the DST (donor specific transfusion) recipients accept heart allografts but reject hyperacutely i.v. infused lymphocytes from the same as DST donor. The present study was devoted to elucidation of the mechanism of these divergent processes. Syngeneic BN hearts and mesenteric lymph node lymphocytes were transplanted to LEW rats pretreated one week previously with donor specific blood transfusions. The allogeneic BN lymphocytes transplanted i.v. to LEW rats receiving one BN DST were rejected hyperacutely within 6 hrs, whereas BN heart grafts transplanted to the BN DST-treated LEW recipients survived 14 +/- 2 days. Adoptive transfer of LEW anti-BN DST sera to naive LEW rats caused destruction of the transplanted BN lymphocytes. The LEW BN DST recipients possessed IgG and IgM class alloantibodies binding to BN lymphocytes and heart endothelial cells. mAbs against MHC class I (OX18) and class II (OX6) antigens neither blocked binding of antibodies of DST-recipient sera to BN lymphocytes nor protected the preincubated BN lymphocytes against destruction after transplantation. Western blot analysis revealed that alloantibodies from DST-recipient sera bound strongly to BN lymphocyte membrane proteins of 60 kd m.w. but not to 45 kd and 30 kd MHC class I and II proteins. Taken together, DST has no protective effect on intravenously transplanted cells. In contrast, it accelerates the rejection. Alloantibodies present in DST-recipient sera "shield" antigens on the surface of organ allograft endothelial cells, thereby protecting them from recognition and cytotoxic effect. Simultaneously, these alloantibodies "opsonize" the intravenously transplanted lymphocytes and facilitate their halting in lymphoid organs and subsequent lysis. Antibodies other than those directed against MHC seem to mediate both these processes. The results of these studies provide also evidence that the effector mechanism of rejection may be different depending on location of the graft, in the lymphoid as in case of transplanted lymphocytes or in non-lymphoid tissues as heart grafts.