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Ventricular assist devices may function as a bridge to recovery or heart transplantation, however, little is known about its mechanisms. This study examined the role of matrix metalloproteinases (MMP)-tissue inhibitors of metalloproteinases (TIMP) axis in the process of recovery after unloading in a rat ischemic-induce heart failure (HF) model. Myocardial infarction model was created with the coronary artery ligation. The infarcted rats hearts were unloaded by heterotopic cardiac transplantation (n=14). 2 weeks later, the function of normal and infarcted hearts with or without loading was evaluated by Langendorff perfusion model. The hearts were then harvested and prepared for the study of expression of MMPs and TIMPs. Developed pressure in the unloading group was higher than the loading group (P=0.0074). Unloading increased the ratio of TIMP-1-MMP-1(1.38±0.11 vs. 0.76±0.09, P<0.05), TIMP-2-MMP-2 (1.06±0.10 vs. 0.33±0.07, P<0.01), TIMP-3-MMP-9(1.07±0.08 vs. 0.59±0.06, P<0.05). Although MMP-1, 2, 9 were downregulated (P<0.01, 0.01, 0.05, respectively), TIMP-2 and TIMP-3 upregulated (P<0.01, 0.05, respectively), MMP-7 and TIMP-1 was not affected significantly. The infarcted cardiac function could be improved by unloading. It was attributed to downregulation of MMP-1, 2 and 9, and upregulation of TIMP-2 and -3, and furthermore, the ratio of TIMPs to MMPs was increased, which might be more sensitive than sole MMPs or TIMPs for the judgment of myocardial matrix homeostasis.

Aim: To investigate the relationship between the expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 and ECM accumulation in rat left ventricle in a mechanical unloaded heart model., Methods: 12-week-old male Lewis rats were subjected to abdominal heterotopic heart transplantation to achieve pressure and volume unloading(mechanical unloading). Age and sex matched in situ heart of Lewis rats were used as control. Collagen volume fraction(CVF) was analyzed by picrosiris-red staining plus polarized microscopy. MMP-2 and -9 gelatinolytic activity were measured by gelatin-zymography. mRNA level of MMP-2, MMP-9, TIMP-1 and TIMP-2 were measured by real-time quantitative PCR. TIMP-1 and TIMP-2 protein level were measured by immunoblotting., Results: Myocardial cross-sectional area of transplanted heart was significantly reduced, and accompanied by excessive ECM deposition (CVF 5.22% +/- 1.6% vs. 2.21% +/- 0.9%, P < 0.05) compared to in situ heart. MMP-2 and MMP-9 activity were significantly increased, as well as mRNA level of MMP-2, MMP-9, TIMP-1 and TIMP-2 compared to in situ heart. TIMP-1 and TIMP-2 protein level in mechanically unloaded heart were significantly upregulated compared to in situ heart, especially for TIMP-1., Conclusion: Mechanical unloading of left ventricle may lead to excessive ECM deposition, accompanied by imbalance between MMPs and TIMPs system, especially the upregulation of TIMPs.

Background: The effect of blocking lymphocyte chemokine receptors with TAK-779 was investigated using a rat intestinal transplantation model., Methods: Dark Agouti rat small intestines were heterotopically transplanted into Lewis rats. The recipients were treated with TAK-779 (10 mg/kg per day) by subcutaneous injection. Graft survival, histologic changes, mixed lymphocyte reaction, and antibody production were examined. Furthermore, expression of the chemokine receptors on the graft-infiltrating lymphocytes in the mesenteric lymph node (MLN) and Peyer's patches (PP) were measured using fluorescence-activated cell sorter analysis., Results: Average duration of survival was greater for group T (with TAK-779: 9.8+/-1.4) than group A (without TAK-779: 7.0+/-0.6). Histologic findings and immunohistochemistry of the graft were consistent with the prolonged survival in group T. In the fluorescence-activated cell sorter analysis, several CD4+ and CD8+ cells were significantly suppressed in the blood, spleen, and MLN of the recipient and in the PP of the graft on postoperative day (POD) 6, but recovered in recipient spleen and MLN by POD 9. However, double-staining of graft-infiltrating lymphocytes in MLN and PP showed a significant reduction in the numbers of T cells expressing CCR5 and CXCR3 by POD 9. On the other hand, mixed lymphocyte reaction and cytokine production, and the antidonor antibody titer were suppressed on POD 6 but not on POD 9., Conclusion: TAK779 diminished not only the number of the graft-infiltrating cells and their expression of CCR5 and CXCR3, but also the total number of recipient T cells that play a role in graft rejection. Further exploration of these effects will provide the novel treatment of the intestinal transplantation.

Objective: We developed a model for intravital microscopic analysis of the coronary microcirculation in transplanted murine hearts and assessed the influence of cold ischemia on postischemic microcirculatory dysfunctions., Methods: Murine hearts were exposed to 60 (n = 12) and 240 minutes (n = 8) of cold ischemia before syngeneic heterotopic transplantation. Intravital fluorescence microscopy allowed detailed analysis of the right ventricular coronary microcirculation, including feeding arterioles, nutritive capillaries, and postcapillary venules. With this technique, we further studied leukocyte-endothelial cell interactions, microvascular permeability, tissue oxygenation, and microlymphatics., Results: Cold ischemia of 240 minutes aggravated nutritive capillary perfusion failure, indicated by a significant reduction of functional capillary density and capillary flow velocity by 63% and 45% (P < .05 vs 60-minute cold ischemic isografts). The mitochondrial redox state, visualized by nicotinamide adenine dinucleotide hydrogen autofluorescence, was markedly deteriorated after 240-minute cold ischemia (P < .05), indicating a persistent mismatch between oxygen supply and demand resulting from pronounced capillary no-reflow. Prolonged ischemia further resulted in 6- and 11-fold higher numbers of rolling and firmly adherent leukocytes in postcapillary venules (P < .05), together with increased microvascular permeability., Conclusions: We introduce a novel approach to visualize in detail the murine coronary microcirculation in vivo by multifluorescence microscopy. Our data demonstrate that prolonged cold ischemia provokes posttransplant capillary no-reflow, leukocytic inflammation, and persistent tissue hypoxia.

Background: Clinical experience with portal vein arterialization (PVA) in liver transplantation is controversial. One reason for this is the lack of standardized flow regulation. The present experiments aimed to establish flow regulation in the arterialized portal vein for heterotopic auxiliary liver transplantation (HALT), to obtain physiological portal blood flow, and to compare this technique with orthotopic liver transplantation., Material/methods: Lewis rats were divided into 7 groups (n = 8 transplantations/group). Group: A I-IV: In HALT, the portal vein was anastomosed to the right renal artery using stents with different diameters (0.2, 0.3, 0.4, 0.5 mm). Afterwards, HALT with PVA using the stent diameter that had achieved the most physiological portal blood flow (group B II) was compared with orthotopic liver transplantation with porto-portal anastomosis (group B III) and to the sham group (B I)., Results: After reperfusion, only the 0.3 mm stent resulted in an average blood flow in the arterialized portal vein in HALT which was within the normal range (1.7+/-0.4 ml/min/g liver weight). The parameters of microcirculation and early graft function were significantly better in group B II than in group B III (functional sinusoidal density: 335+/-48 vs. 224+/-31/cm, diameter of sinusoids: 6.4+/-0.6 vs. 5.2+/-0.6 microm, diameter of postsinusoidal venules: 31.1+/-3.3 vs. 25.5+/-2.0 microm, bile-production: 27+/-8 vs. 19+/-5 microl/h/g liver weight)., Conclusions: Using an optimal stent diameter in HALT with portal vein arterialization, an adequate flow-regulation can be achieved. Avoiding portal hyper- and hypoperfusion, good results for microcirculation and early graft function can be obtained.

We report 3 patients who initially underwent heterotopic transplantation due to a size mismatch but then later underwent orthotopic retransplantation because of heart failure. In each case, the heterotopic graft was left in place, the native heart was removed, and the new allograft was placed orthotopically. This technique resulted in reduced postoperative morbidity and excellent long-term outcomes.

Clinical islet transplantation in liver has achieved normoglycemia. However, this site may not be ideal for islet survival. To create a more optimal site for islet transplantation, we have developed a construct with biodegradable scaffolds. Islets were seeded in scaffolds and transplanted into the epididymal fat pad of diabetic BALB/c mice. Controls included islets transplanted underneath the kidney capsule or into the fat pad without scaffolds. All animals with islets in scaffolds or the kidney became normoglycemic and maintained this metabolic state. When islets were transplanted without scaffolds the time to achieve normoglycemia was significantly increased and less than 45% of mice survived. An oral glucose tolerance test was performed on the scaffold and kidney groups with similar blood glucose levels and area under the curve values between the groups. Grafts were removed at more than 100 days posttransplantation and all animals became hyperglycemic. There was no significant difference in insulin content between the grafts and all grafts were well vascularized with insulin-positive beta cells. Therefore, islets in scaffolds function and restore diabetic animals to normoglycemic levels, similar to islets transplanted underneath the kidney capsule, suggesting scaffolds can be used to create a site for islet transplantation.

Background: The utility of combining strategies of myocardial protection was studied in intact rat hearts subjected to 1 hour of ischemia and 40 minutes blood reperfusion., Methods: Lewis rats (n = 48) were divided into 4 transplant groups. Twenty-four hearts were arrested by coronary perfusion with hypothermic Celsior solution at 60 mm Hg. The aortic valve was punctured to introduce volume into the left ventricle (LV), and the hearts were abdominally isografted. Animals were either given both the antioxidant probucol (300 mg/kg) and the sodium-hydrogen exchange inhibitor cariporide (5 mg/kg) (CP; n = 6), just cariporide (CAR; n = 6), just probucol (PROB; n = 6), or neither drug (CON; n = 6). After 40 minutes of blood reperfusion, transplanted hearts were rearrested. The control recipients' native hearts (native; n = 6) were also arrested. Postmortem LV compliance relations and myocardial water content (MWC) were measured., Results: Grafts protected by probucol were significantly more compliant than controls and significantly less compliant than grafts protected by cariporide alone and with both cariporide and probucol (p = 0.0001, analysis of variance). Compliance relations for CP overlapped those for CAR. All grafts were less compliant than natives. MWC was significantly greater in controls and PROB than in natives., Conclusions: Pretreatment with cariporide in the setting of ischemia-reperfusion injury provides greater protection against the development of diastolic abnormalities than probucol when Celsior solution is used for both arrest and preservation. In this model, there is no advantage to combining the drugs, supporting the hypothesis that there is an overlapping mechanism of protection.

The purpose of the study was to determine the feasibility of preserving ovarian function after heterotopic transplantation by means of microvascular anastomosis of the transplanted vascular pedicles to a set of preselected vessels. Six groups of 10 Sprague-Dawley inbred rats were used in this study. Group I underwent bilateral ovariectomy operation and served as the ovariectomy control. Group II underwent bilateral ovariectomy followed by heterotopic isogenic ovarian implantation. Group III underwent bilateral ovariectomy and isogenic heterotopic ovarian transplantation by means of microvascular anastomosis. Group IV served as the laparotomy sham-operated control. Group V served as the ovarian donor for group II. Group VI served as the donor of the ovarian-kidney vascular pedicle complex for group III. Postoperative ovarian estradiol levels were measured, and histological characteristics were elucidated in groups I, II, III, and IV. The results demonstrated that the estradiol level of the transplantation group was comparable to that of the sham operation group and was significantly higher than that of the implantation group. Histologically normal ovarian architecture was observed in the sham group (IV) and also in the transplantation group (III). Altered architecture was observed in the implantation group (II). These findings indicate that extraabdominal heterotopic ovarian transplantation with microvascular anastomosis led to normal ovarian hormonal function and was effective in preserving oocyte production capacity.

Objective: To evaluate the role of inhibin in elevated base levels of FSH and follicular hyperplasia in ovarian autotransplantation in rats., Design: Experimental animal study., Setting: Unit of Experimental Research at the Barcelona University School of Medicine., Animal(s): Female syngeneic Lewis rats aged 16 weeks., Intervention(s): The animals were randomized into two groups: group A, control group undergoing only laparotomy (n = 5) and group B, oophorectomized with SC autologous heterotopic transplant (n = 5). The animals were killed and their ovaries removed for histologic, morphometric, and immunohistochemical analysis at 28 days after surgery in both groups., Main Outcome Measure(s): Serum levels of E2 and FSH were determined on day 0 (the day of surgery or baseline) and days 4, 7, 14, 21, and 28. Morphometric analysis of ovarian structure for evaluation of antral follicles and their granulosa cell area and immunohistochemistry for inhibin staining were also done., Result(s): The endocrinological function recovered at 28 days, and the FSH levels for the transplant group were significantly higher than for the group with normoinsert ovary. Morphometric analysis showed that the mean granulosa cell area was greater in group B when compared with the control group. Immunohistochemistry revealed almost null inhibin staining of the stroma in transplanted ovarian tissue., Conclusion(s): Tissue damage brought on by ischemia in the transplant of nonvascularized ovaries may bring about an inhibin deficit in the ovarian stroma, which might explain the increased levels of FSH. These increased levels, in turn, would be responsible for the follicular hyperplasia seen in this tissue when it recovers its function.

Obliterative bronchiolitis (OB) is the most important cause of graft dysfunction post-lung transplantation. It is likely that the small airway epithelium is a target of the alloimmune response, and that epithelial integrity is a crucial determinant of airway patency. Our goals are to elucidate epithelial cell kinetics in the heterotopic mouse trachea model and to determine potential mechanisms of cell death in allografts. Allografts and isografts were obtained by transplanting BALB/c tracheas into C57BL/6 and BALB/c immunosuppressed and non-immunosuppressed hosts, respectively and harvested from day 3-20. Morphometry, BrdU and TUNEL labeling, and EM studies were performed. Columnar epithelium in isografts and allografts sloughs during day 0-3, but regenerates in both sets of grafts by day 10. Subsequently, allografts become inflamed and denuded, while isografts retain an intact epithelium. Prior to airway denudation, allografts exhibited significantly increased epithelial cell density, BrdU labeling index (LI), and TUNEL positive cells. Epithelial apoptosis was confirmed by electron microscopy. Allograft percent ciliated columnar epithelium and lumenal circumference were significantly decreased. Cyclosporin delayed airway fibrosis but did not alter the progression of the allograft through the phases of early ischemic injury, airway epithelial cell regeneration, and eventual cell death. These studies quantitatively demonstrate that the allograft epithelium actively regenerates in the alloimmune environment, but succumbs to increased apoptotic cell death, underscoring the importance of the airway epithelium as a self-renewing source of alloantigen.

Background: The Th1-Th2 paradigm proposes clonal expansion of Th2 lymphocytes as the basis of allograft tolerance. The Th2 cells have been found to be present in recipients with long-term allograft survival. However, the presence of Th2 cells and tolerance is not a uniform finding. Previously we have shown that pre-engraftment single dose rapamycin and a 7-d course of cyclosporin induce transplantation tolerance to 120 d. In the present study, we investigated the immunobiology of grafts in a long-term follow-up (>350 d)., Methods: Kidney allografts (n = 7), isografts (n = 5) and single nephrectomy (n = 3) groups were followed for 350 +/- 87 d. Heterotopic kidney transplant was performed by the same surgeon in the allograft group (ACI-Lewis) and the isograft group (Lewis-Lewis). The left kidney was removed in the single nephrectomy group. The allograft group was treated with pre-engraftment single dose rapamycin and a 7-d course of cyclosporin. A kidney biopsy was performed at midpoint time for histological study and tissue was frozen for measuring intragraft cytokine expression (IL-4, IL-10) in all animals. Prior to biopsy, serum blood urea nitrogen (BUN) and creatinine (Cr) levels were studied. Serum BUN, Cr levels, plus 24-h urinary protein (PRO) were measured prior to sacrifice. Randomly, four allograft rats received skin grafts (ACI, Lewis and Buffalo skin donors) after kidney biopsy. Skin grafts were studied for a mean of 6 weeks for signs of acceptance or rejection. Analysis of variance (ANOVA) with Tukey's test was used; p < 0.05 was considered statistically significant., Results: The mean follow up was 352 +/- 87 d. BUN and Cr levels at biopsy time (mean 214 d) were not statistically different between the three groups (p = 0.19 and p = 0.66). At sacrifice (mean 352 d), BUN, Cr and PRO were statistically different between allograft and isograft groups (p = 0.013), and between allograft and single nephrectomy groups (p = 0.027). Functional and histological signs of graft loss occurred in three of seven (42.8%) of the allografts at 352 d. Using BANFF criteria, three allografts at biopsy time and seven allografts (100%) and four isografts (80%) at sacrifice time developed chronic histologic changes. Intragraft overexpression of IL-4 and IL-10 was seen at biopsy and sacrifice time in six of seven allografts and one of five isografts. All donor specific skin grafts (ACI-Lewis) on allografts were accepted and third party (Buffalo) donor skin grafts were rejected in all animals (>95% skin necrosis)., Conclusions: This highly stringent, functional, renal transplant model yields 100% normal renal function as compared with isografts at 120 d follow-up. With the follow-up extended to 350 d, 43% of the allografts loose function and develop a chronic allograft histology despite a demonstrated intragraft Th2 cytokine dominance and donor specific skin graft acceptance.

We took advantage of the combination of a rat heart transplantation model with a modified differential display RT-PCR method to identify transcriptome changes in the right atria from transplanted compared with native hearts. Based on sequence homology search, the 37 cDNAs differentially displayed both 2 and 7 days posttransplantation were categorized into 7 unknown transcripts, 16 expressed sequence tags (ESTs), and 14 partially or completely characterized genes. The last group cDNAs, validated by relative RT-PCR, belonged to diverse gene families involved in specific metabolisms, protein synthesis, cell signaling, and transcription. Furthermore, we identified differential transcripts corresponding to denervation and fetal gene reexpression. We found coordinate downregulation of genes involved in energy metabolism and protein synthesis regulation, similar to that reported for senescent skeletal muscle. From these transcriptome changes, we propose that heart transplants and senescent muscles share common molecular mechanisms.

The transplantation of ovarian tissue has recently been the focus of intense investigation with the aim of avoiding premature ovarian failure mainly in patients receiving chemotherapy or radiotherapy for malignant disease. Here, we present an evaluation of the long-term function of both fresh (patients 1, 2, and 3) and cryopreserved (patient 4) ovarian autografts in four premenopausal patients aged 46-49 yr who underwent heterotopic ovarian transplantation and were followed over a 1-yr period without receiving gonadotropins to stimulate follicular growth. In patients 1 and 2, approximately 1 cm(3) ovarian cortical autograft was placed sc in the inner aspect of the arm, whereas and in patients 3 and 4 minced ovarian tissue was placed into a muscle pocket in the abdominal wall. In patients 1, 2, and 4 the ovarian hormone secretion (as suggested by sequential estradiol and FSH serum measurements) was reestablished 3-4 months after autotransplantation, and graft function was not improved by immediate rather than delayed heterotopic ovarian autografting. Despite a reestablished ovarian function, a 2- to 7-fold increase in peripheral FSH concentration was evidenced. The cases reported here suggest that hormonal protection can be restored after fresh or cryopreserved heterotopic ovarian transplantation in women, albeit for only a short reproductive span.

Revascularisation of bone grafts is influenced by both the anatomical origin and the pre-implantation processing of the graft. We investigated the revascularisation by entrapment of 141Ce (cerium)-labelled microspheres in large, fresh and demineralised syngeneic grafts of predominantly cancellous (iliac bone) or cortical (tibial diaphysis) bone three weeks after heterotopic implantation in rats. The mean (SD) 141Ce deposition index (counts per minute (cpm) of mg recovered implant/cpm of mg host iliac bone) was higher in fresh iliac bone grafts, 0.98 (0.46) compared to that of demineralised iliac bone, 0.32 (0.20), p < 0.001, and fresh tibial bone grafts, 0.51 (0.27), p = 0.007. We found no significant difference in the mean 141Ce deposition index between fresh tibial bone grafts and demineralised tibial bone grafts, 0.35 (0.42), p = 0.4, or between demineralised tibial grafts and demineralised iliac bone grafts, p = 0.8. The results suggest that whereas fresh cancellous grafts are revascularised more completely than fresh cortical grafts, there is no difference in the revascularisation of demineralised cancellous and cortical grafts. In addition, fresh cancellous bone is revascularised more completely than demineralised cancellous bone, whereas there is no difference between fresh and demineralised cortical bone.

Objective: In acute cardiac rejection, changes in myocardial oxygen consumption occur; non-invasive detection of these metabolic changes would have obvious clinical utility. In the classic cervical, heterotopic, canine, transplant model, the heart is non-working. It has a low myocardial oxygen consumption. Creation of a working model with normal myocardial oxygen consumption would enhance validity of non-human studies., Methods: Clearance of 11C acetate was determined by positron emission tomography (PET) scanning and compared with myocardial oxygen consumption in normal and transplanted canine hearts. Donor hearts from mongrel dogs (2.5-3 kg; n=4) were transplanted into the neck of adult beagles (12-15 kg; n=4), no immunosuppression was given. Two non-working hearts were modified to eject only coronary flow via the right ventricle. In two hearts, a novel working model was created with aortic regurgitation to load the left ventricle. Working and non-working hearts underwent PET scanning on post-operative days 2 and 4. Normal dog hearts (n=2) and native hearts of transplanted dogs (n=3) were used to validate the scanning technique. Coronary sinus and aortic oxygen saturation data along with myocardial blood flow (radiolabeled microspheres) confirmed that clearance of 11C acetate in normal and transplanted hearts followed a bi-exponential model., Results: Myocardial oxygen consumption was correlated with the rate constant of 11C acetate rapid phase clearance (r=0.91) in normal and transplanted hearts. The working hearts had increased myocardial oxygen consumption compared to non-working hearts., Conclusions: This study (1) introduces a model of a working heterotopic cardiac transplantation with near-normal oxygen consumption; and (2) demonstrates that regional myocardial oxygen consumption in transplanted hearts can be detected by 11C acetate PET.

Background: The widely used non-volume-loaded abdominal heterotopic heart transplant (NL) in rats undergoes atrophy after transplantation. Various techniques have been designed to load the transplanted heart because of its potential immunological impact. Our aim was to create a volume-loaded heterotopic heart transplantation model (VL) capable of ejection and practical for routine studies. Using this model, we tested the hypothesis that VL isografts would retain myocardial performance comparable to native hearts (NH)., Methods: Heterotopic hearts were transplanted using and end-to-side anastomosis between the donor's superior vena cava and the recipient's abdominal inferior vena cava. The right ventricle loads the left ventricle (LV) via a direct anastomosis of the pulmonary artery to the left atrium. The LV ejects volume through an end-to-side anastomosis of the donor's aorta to the recipient's abdominal aorta. Hemodynamic data (systolic and diastolic LV pressures, dP/dt max and min, tau) were studied in-situ (at baseline and after adding volume) and in a Langendorff perfusion system (at baseline and after stimulation with isoproterenol) 2 weeks after transplantation., Results: In situ systolic pressure and diastolic function of VL was superior to NL, and beta-adrenergic stimulated performance in the Langendorff perfusion of VL showed hemodynamic performance equivalent to NH, unlike NL which had a diminished response., Conclusion: This technique results in a volume-loaded ejecting heart transplant model that preserves anatomical structures. The VL can be evaluated in situ and after explantation in Langendorff perfusion system and may offer advantages if workload of the graft is of significance to the study performed.

Background: A less-invasive method for cardiac allograft surveillance than endocardial biopsy is needed. We analyzed heart rate variability of heterotopically transplanted rat hearts as a method of detecting rejection of rat cardiac allografts., Methods: Two kinds of heterotopic transplants were performed: 1) Brown-Norway rats received Brown-Norway rat isografts, and 2) Lewis rats received Brown-Norway rat allografts. The electrocardiogram (ECG) of the grafts were serially recorded under non-anesthetized and non-restricted conditions using a telemetric ECG transmitter implanted in the recipient's abdomen. Frequency domain analysis of the ECGs was performed using a fast Fourier algorithm., Results: Total power of the heart rate variability in the isograft heart was reduced to 1.1%, compared to normal subjects without transplantation (p < .001). In the allograft heart, it was also reduced to 1.0% on days 1.5 (rejection score 0 to 1), but gradually increased thereafter up to 185% on day 6 (rejection score 3.75+/-0.50). The increase in spectral power was frequency-dependent (i.e., changes in the power in lower frequency range [LF, 0.04 to 0.67 Hz] were significantly higher than other ranges). This increase was reversible when immunosuppressive therapy was performed with the use of cyclosporine A. In the allograft group, peak-to-peak amplitudes of the QRS complex and heart rate were significantly decreased on day 5.5 or later, whereas the power of the LF was significantly increased by day 3.5 or later., Conclusions: Our data suggest that heart rate variability analysis is a promising noninvasive marker for early detection of cardiac allograft rejection. This method may also provide a sensitive means of assessing the effects of immunosuppressive therapy.

Background: Limitations of the dog model of orthotopic heart transplantation to study rejection include the need for extracorporeal circulation and transfusions. Heterotopic cervical heart transplantation may improve on these limitations. It is not known whether the natural history after heterotopic transplant is similar to that after orthotopic grafting., Methods: Twenty-one dogs underwent cervical heterotopic heart transplantation. Serial echocardiographic studies were performed 1 to 3 hours after surgery, at 24 hours, 48 hours later, and immediately before killing (5 to 7 days)., Results: LV diastolic and systolic areas were elevated immediately after transplantation (4.95+/- 1.49 cm2 and 3.36+/-1.18 cm2 respectively) but decreased at 24 hours (3.93+/-1.20 cm2, p = 0.0003 and 2.44+/-0.96 cm2, p = 0.16). Thereafter, a progressive increase in LV diastolic and systolic areas was observed until sacrifice (5.53+/-2.20 cm2 and 4.59 +/-2.14 cm2, p < 0.001 vs 24 hours). LV fractional area shortening (FAS) and fractional volume change were depressed immediately after transplantation (28.2+/- 12.8% and 40.4+/-12.3%, respectively), but increased at 24 hours (35.7+/-10.0%,p = 0.11 and 50.3+/-4.0%,p = 0.02). FAS decreased at 48 hours to 19.6+/-11.1% (p = 0.01 vs 24 hours). The centractility indexes were markedly reduced before killing (FAS = 14.0+/-8.2% and LVEF = 18.4+/-1.3%, p < 0.0005 vs 24 hours). The thickness of the interventricular septum increased from 11.9+/-2.0 mm at baseline to 14.4+/-4.2 mm before sacrifice (p = 0.007)., Conclusion: The evolution of dogs after heterotopic cervical heart transplant is comparable to that after the more standard orthotopic graft. Considering its multiple practical advantages including the easy echocardiographic follow-up, heterotopic transplantation may become a very practical model to use for the study of rejection after heart transplantation.

Background: Changes in the monophasic action potential may be used for detecting early acute rejection in the transplanted rat heart., Methods: Heterotopic heart transplantations were performed in allogeneic and syngeneic rats. During atrial pacing, monophasic action potentials were simultaneously recorded in the right atrium and ventricle of the transplanted hearts on postoperative days 1, 3, and 5. The amplitude and duration of monophasic action potentials, atrioventricular conduction time, and cardiac intervals were analyzed. Histopathologic examination for rejection was performed on postoperative days 1, 3, and 5., Results: In the allogeneic group, monophasic action potential amplitude progressively decreased, and monophasic action potential duration gradually increased after heart transplantation in the atrium and ventricle. With rejection, the amplitude decreased to a greater extent, and monophasic action potential duration increased to a greater extent in the atrium than in the ventricle on day 3. The atrioventricular conduction time increased on day 5, but the cardiac interval did not change. An inverse correlation between histopathologic grade and the monophasic action potential amplitude, and a positive correlation between histopathologic grade and the monophasic action potential duration existed for both the atrium and ventricle. These electrophysiologic and histopathologic changes were not observed in the syngeneic group., Conclusions: We conclude that right atrial monophasic action potentials may be a useful and reliable indicator of early acute heart transplant rejection.

Background: Current clinical heart preservation is still limited to 6 hours. A suitable heart transplantation model to rapidly screen the effectiveness of new solutions is essential. This study examines a new screening test-a modification of the conventional abdominal rat heart transplantation model that overcomes its serious limitation of lack of quantitative evaluation of function., Methods: Rat hearts, with an externalized intraventricular balloon-tipped catheter, were transplanted immediately (controls) or flushed and stored in colloid-free University of Wisconsin solution in ice for 6, 9, or 12 hours before transplantation. One and 7 days later this catheter was connected to a pressure transducer and a calibrated syringe. Heart rate, maximum developed pressure, and maximum rate of left ventricular pressure rise were determined. Grafts were prepared for histologic study on day 7., Results: All preserved hearts commenced beating within 2 minutes (controls beat within 20 seconds). On day 1 the heart rate and chamber stiffness (deltaP/deltat) were similar in all groups. The 9- and 12-hour-preserved hearts had significantly (p < 0.05) diminished developed pressure and contractility. On day 7 contractility and developed pressure improved in 9- and 12-hour-preserved grafts. There was extensive muscular atrophy and necrosis, with extensive cellular infiltrate in the 9- and 12-hour-preserved grafts; other grafts showed no damage., Conclusion: This quantitative model provides an ischemia-related gradation of function and greater discrimination than conventional methods. It has refuted previous studies suggesting effective preservation for 20 hours and demonstrated that functional testing is essential in evaluating preservation regimens.

Islet allografts in insulin-dependent diabetic (IDDM) patients exhibit variable survival lengths and low rates of insulin-independence despite treatment with anti-T-cell antibodies and maintenance immunosuppression. Use of poorly characterized freshly isolated preparations makes it difficult to determine whether failures are caused by variations in donor tissue. This study assesses survival of standardized beta-cell allografts in C-peptide negative IDDM patients on maintenance immunosuppression following kidney transplantation and without receiving anti-T-cell antibodies or additional immunosuppression. Human islets were isolated from pancreatic segments after maximal 20 h cold-preservation. During culture, preparations were selected according to quality control tests and combined with grafts with standardized cell composition (> or = 50% beta cells), viability (> or = 90%), total beta-cell number (1 to 2 x 10(6)/kg body weight) and insulin-producing capacity (2 to 4 nmol x graft(-1) x h(-1)). Grafts were injected in a liver segment through the repermeabilized umbilical vein. After 2 weeks C-peptide positivity, four out of seven recipients became C-peptide negative; two of them were initially GAD65-antibody positive and exhibited a rise in titre during graft destruction. The other three patients remained C-peptide positive for more than 1 year, two of them becoming insulin-independent with near-normal fasting glycaemia and HbA1c; they remained GAD65- and islet cell antibody negative. The three patients with surviving grafts presented a history of anti-thymocyte globulin therapy at kidney transplantation. Long-term surviving grafts increased C-peptide release following intravenous glucagon or oral glucose but not following intravenous glucose. Thus, cultured human beta-cells can survive for more than 1 year in IDDM patients on maintenance anti-rejection therapy for a prior kidney graft and without the need for an increased immunosuppression at the time of implantation. The use of functionally standardized beta-cell grafts helps to identify recipient and graft factors which influence their survival and metabolic effects. Insulin-independence can be achieved by injection of 1.5 million beta-cells per kg body weight in a liver segment. These beta-cell implants respond well to adenylcyclase activators but poorly to glucose.

Background: Intestinal grafts are placed either heterotopically (out of continuity) or orthotopically (in continuity); the latter is believed to be advantageous, as intraluminal nutrients and intestinal secretions might modulate the intestinal immune status and possibly delay rejection., Methods: This study was designed to delineate the effects of heterotopic versus orthotopic allograft position on the morphology and function of intestinal smooth muscle in our rat model of chronic rejection. Syngeneic orthotopic grafts were evaluated to control for changes due to the transplantation process., Results: Histochemistry of the graft's muscularis externa showed a significant thickening due to hyperplasia and hypertrophy, which was most pronounced in heterotopic grafts (control = 92+/-2.4 microm, syngeneic grafts = 140+/-6.7 microm, orthotopic allografts = 278+/-26.6 microm, heterotopic allografts = 456+/-50 microm). In terms of function, muscle strips from allografts only generated 23% of the total bethanechol-induced contractile force in vitro compared to unoperated controls and syngeneic grafts. The mean resting membrane potential of control and isograft muscle cells was -69 +/- 0.9 mV with a slow-wave amplitude of 20+/-0.5 mV. Chronic rejection hyperpolarized the resting membrane potential of orthotopic allografts (-66 +/- 0.5 mV) and even more so of heterotopic allografts (-58 +/- 3.4 mV). Slow-wave amplitudes were decreased in orthotopic (14+/-0.9 mV) and nearly abolished in heterotopic allografts (2+/-1.2 mV)., Conclusions: Our data indicate that allografts in heterotopic position are most susceptible to the insult of chronic rejection exemplified by increased proliferative and hypertrophic transformation of intestinal smooth muscle and a marked decrease in mechanical and electrical activity.

A 30-year-old man with dilated cardiomyopathy underwent heterotopic heart transplantation, concurrently with surgical reduction of the native left ventricle, a mitral valvuloplasty, and an apical thrombectomy. Postoperatively, the patient had hemodynamic instability that was refractory to conventional medical therapy. Temporary sequential pacing of the 2 hearts was used to induce timed counterpulsation, which resulted in marked hemodynamic improvement. To the best of our knowledge, this case involves the 1st report in the English-language literature of clinical usage of paced, linked counter-pulsation to treat acute hemodynamic compromise early after heterotopic heart transplantation.

The revascularization of islets of Langerhans transplanted in heterotopic sites like the liver by portal vein embolization or the renal subcapsular space is a major process necessary for the viability of grafted cells. This process has been extensively studied by different techniques and the results have shown that islet revascularization is an early phenomenon that takes place soon after transplantation. In this report we have analyzed by a double indirect immunofluorescence technique, the revascularization process of purified endocrine islet beta-cells transplanted in the renal subcapsular space of syngeneic rats. Lewis rats were grafted with islets cultured for 24 h, with a suspension of purified beta-cells cultured for 24 h, and with a suspension of purified beta plus nonbeta-cells cultured for 24 h. Rats were killed at different days after implantation and the kidney bearing the grafts were snap frozen and immunohistochemically stained with a rabbit anti factor VIII antiserum (which labels endothelial cells). Immunocytochemical analysis revealed that cultured islets completed revascularization by days 3-5 after transplantation, as shown by the detection of capillary endothelial cells within and surrounding the islets. Within purified endocrine beta-cell grafts, the presence of numerous endothelial cells was not observed until days 10-14, indicating that revascularization of beta-cells with host vessels is not such an early phenomenon as it takes place in whole isolated islets. Conversely, the addition of a population of endocrine nonbeta-cells to the purified islet cell grafts, partially accelerated the revascularization of pure beta-cell grafts, which showed the presence of abundant capillary endothelial cells already at day 7 after transplantation, indicating that some other unidentified factors besides the absence of endothelial cells may explain the retardation of beta-cell grafts revascularization.

The role of the thymus in the ongoing acquisition of tolerance to self antigens has made it an attractive site for islet transplantation. Several studies have reported survival of rodent islet allografts in the thymus without requiring the long-term use of immunosuppressive agents; however, the degree of glucose homeostasis in the intrathymic islet transplant recipients has not been examined. We transplanted 500, 1000, or 2000 syngeneic islets into the thymus of streptozotocin-induced diabetic Wistar-Furth rats, and compared the metabolic response of these recipients with animals receiving 2000 syngeneic islets under the kidney capsule. Three of four recipients which received 2000 islets under the kidney capsule achieved normoglycemia (< or =8.4 mmol/L) within 1 wk and all animals became normoglycemic within 2 wk posttransplantation. In contrast, intrathymic implantation of 2000 islets induced normoglycemia in only one of six recipients during the same time interval, and when this number was reduced to 1000 or 500 islets, none of the recipients (n = 6) normalized within 1 wk posttransplantation. Animals that received an intrathymic transplant were glucose intolerant compared to normal controls and animals with subcapsular islet transplant. Removal of the graft-bearing organs resulted in hyperglycemia in all cases, and examination of the grafts revealed the presence of numerous well-granulated insulin-containing cells in both sites. The cellular insulin content of the subcapsular grafts (67.4 +/- 12.1 microg; n = 4) was significantly higher (p < or =0.05) than what was extracted from intrathymic grafts (9.5 +/- 1.2 microg from 1000 islets; n = 3 and 20.0 +/- 4.6 microg from 2000 islets; n = 3). We conclude that 2000 syngeneic islets implanted either in the thymus or beneath the kidney capsule can normalize hyperglycemia in streptozotocin-diabetic rats; however, normal glucose tolerance was not established in intrathymic islet recipients, suggesting that a higher number of islets may be necessary to achieve normal glucose homeostasis.

We have demonstrated the utility of ultrasound backscatter microscopy for targeted intraparenchymal injections into embryonic day (E) 13.5 mouse embryos. This system has been used to test the degree of commitment present in neural progenitors from the embryonic ventral telencephalon and mid-hindbrain region. Many E13.5 ventral telencephalic progenitors were observed to integrate and adopt local phenotypes following heterotopic transplantation into telencephalic or mid-hindbrain targets, whereas mid-hindbrain cells of the same stage were unable to integrate and change fate in the telencephalon. In contrast, many mid-hindbrain cells from an earlier developmental stage (E10.5) were capable of integrating and adopting a forebrain phenotype after grafting into the telencephalon, suggesting that mouse mid-hindbrain progenitors become restricted in their developmental potential between E10.5 and E13.5.

Background: Alcohol has potential deleterious effects on donor heart function. This study was conducted in rats to determine whether long-term alcohol ingestion produces impaired hemodynamic performance while maintaining a normal left ventricular ejection fraction in donor hearts before transplantation and whether donor cardiac function is affected after heart transplantation., Methods: Rats fed 30% alcohol in their drinking water for 12 weeks were compared with rats fed a normal diet. Left ventricular ejection fraction was measured by echocardiography with Simpson and single plane Dodge formulas in living sedated rats after 10 and 12 weeks of alcohol feeding. Explanted heart function was assessed before and 3 days after heterotopic heart transplantation (no immunosuppression) with a Langendorff preparation., Results: Blood ethanol levels at 4 and 8 weeks were 0.08 +/- 0.04 and 0.08 +/- 0.09 gm/dl. Left ventricular ejection fraction was similar in the group fed an alcohol diet for 12 weeks when compared with the control group (65.4% +/- 1.6% vs. 66.5% +/- 2.9%, p = 0.33). Explanted alcohol-fed hearts before transplantation had significantly lower maximum and developed pressures and had a blunted response to 0.1 ml 10(-9) mol/L isoproterenol. After transplantation alcohol-fed hearts had significantly lower maximum and developed pressures and decreased maximum rates of pressure rise and pressure decline. Allografts (ACI to Lewis) exhibited decreased function in comparison with isografts (ACI to ACI)., Conclusions: Alcohol feeding for 12 weeks in rats does not affect pretransplantation left ventricular ejection fraction, but it impairs explanted heart function, both before and after transplantation, resulting in a subclinical cardiomyopathy that is worsened by the presence of allograft rejection. Long-term alcohol exposure and rejection have independent, additive detrimental effects on left ventricular performance of the transplanted heart. Alcohol-exposed hearts may not be suitable donors.

Background: Endothelial dysfunction is one of the early events leading to atherosclerosis. It occurs early after orthotopic heart transplantation and precedes the appearance of accelerated graft coronary artery disease believed to stem from chronic rejection of the endothelium. Acute rejection may contribute to the development of graft vasculopathy., Methods: To assess the time course and specific mechanisms of coronary endothelial dysfunction in acute untreated rejection, a swine model of retroperitoneal heterotopic heart transplantation was used. Large white swine (age 10 +/- 2 weeks, weight 25 +/- 5 kg) were serum-typed for class I antigen of the swine leukocyte antigen system and selected to ensure a similar degree of incompatibility. Donor hearts were preserved with normothermic blood cardioplegia and regional hypothermia; the mean ischemic time was 64 +/- 15 minutes. Myocardial contractility decreased from day 5 (normal) to day 14 (weak), but electrical activity was preserved. All coronary arteries were patent, and International Society for Heart and Lung Transplantation grade 4 rejection was present in all hearts beyond 5 days. The endothelial function of epicardial coronary arterial rings of native and transplanted hearts was studied in organ chambers filled with modified Krebs-Ringer bicarbonate solution and compared 1, 5, 9, and 14 days after transplantation., Results: Maximal endothelium-independent relaxations were unaffected at all stages. Endothelium-dependent relaxations to serotonin and alpha 2-adrenergic agonist UK 14304 (which activate receptors coupled to Gi-proteins) and to sodium fluoride (a direct G-protein activator) deteriorated progressively over time. At 14 days maximal relaxations to the calcium ionophore A23187, adenosine diphosphate, and bradykinin were also reduced, but to a lesser degree than those to serotonin and sodium fluoride. Histomorphometric studies of the allograft coronary artery rings showed progressive intimal hyperplasia from day 5 to day 14, with an increase in the incidence from 29% +/- 8.3% to 61.5% +/- 12%., Conclusions: These studies show that endothelial dysfunction in untreated acute rejection after heart transplantation develops beyond 5 days and initially involves G-proteins; the dysfunction worsens over time to finally affect all endothelial mechanisms and vascular smooth muscle. The progression of the associated intimal hyperplasia parallels the alteration in endothelial function, suggesting a permissive role of the dysfunction in the development of this acute form of coronary graft vasculopathy.

Findings of the study on the dog hypogastric nerve regeneration after implantation in caudal large intestine distal region are presented. Not only occurrence of the implanted nerve structure repair was shown, but also its assistance to formation of a well-developed neurovascular plexus in large intestine wall. Regenerated nerve fibres from the nervous plexus that forms in the epineurium of the implanted nerve invade the wall of large intestine. These data indicate the formation of an intramural nervous apparatus in large intestine wall providing additional neurotrophic regulation of an organ.

The purpose of this study was to determine if graft viability during simple cold storage can be monitored from alteration of myocardial electrical impedance. Six anesthetized dogs underwent rapid cardiac extirpation and were placed in simple cold storage in saline for 12 hours. Myocardial electrical impedance was measured serially by use of a LCR meter and the changes of myocardial resistivity analyzed. Myocardial specimens were taken for myocardial ATP analysis and electron microscopic study during preservation. We investigated the correlation between myocardial resistivity and myocardial ATP content. Moreover in order to predict cardiac function after simple cold storage in saline, heterotopic cardiac transplantation in the neck was performed in mongrel dogs and left ventricular Emax (LV Emax) was measured with a micromanometer and conductance catheter method. It was then investigated whether or not it is feasible to predict cardiac function of the graft afer reperfusion from the changes in myocardial resistivity. Myocardial ATP remained above 50% of preischemic value 4 hours after preservation. Ultrastructural alterations of ischemia were observed in hearts preserved for 8 and 12 hours. In heterotopic cardiac transplantation, LV Emax at 120 minutes after reperfusion recovered to 94 +/- 13% of preischemic function in 4 hour-preserved heart, 72 +/- 10% in 8 hour-preserved heart. Percent recovery of LV Emax in the former was significantly higher than that in the latter (p<0.05). Turning points from reversible conditions to irreversible ones were between 4 and 8 hours based on myocardial ATP content, cardiac function after transplantation and morphological changes. With the time of preservation, resistivity began to increase in every dog and peaked during preservation. The time required to reach the peak point of resistivity ranged from 4 to 5.5 hours. Resistivity increase rate (RIR) decreased gradually during preservation and it was over 0.1 omega cm/min until a half-life of ATP. These results suggested that measurement of myocardial electrical impedance in the preserved heart should be feasible as an indicator of graft viability during preservation in heart transplantation.

The current shortage of donors for allotransplantation has generated interest in the potential use of animal organs to meet increasing clinical transplant needs, ie, xenotransplantation. However, when phylogenetically distant species such as the pig are transplanted into unmodified primate hosts--discordant xenotransplantation--the grafts undergo a rapid rejection process characterized by edema, hemorrhage, and diffuse microvascular thrombosis. "Hyperacute rejection" as such is mediated by an IgM natural antibody directed against the galactose alpha(1,3) galactose epitope expressed on the endothelial cell surface of all mammals except old world monkeys, apes, and humans which collectively lack the galactosyltransferase enzyme necessary for antigen expression. Transplants between nonhuman primates and human recipients--concordant xenotransplantation--avoid hyperacute rejection, but nonetheless undergo "acute vascular rejection" and progressive microthrombotic injury. Acute vascular rejection is associated with endothelial cell "activation," loss of vascular integrity, and progressive thrombosis. Molecular strategies for avoiding xenograft rejection involve insertion of genes into the donor pig genome capable of modifying xenoreactive antigen expression and regulating antibody-mediated endothelial cell damage.

Animal studies have been used to investigate different transplantation (Tx) sites to find best conditions for Tx in humans. A few long-term experiments of comparisons of different Tx-sites have been published. Therefore the aim of our study was to compare islet grafts transplanted under the kidney capsule (KTx) of male Lewis rats with grafts transplanted under the spleen capsule (STx) and to observe these animals over a period of 6 months. Diabetes was induced by Streptozotocin and rats were each transplanted with 2000 syngeneic islets under the kidney respectively the spleen capsule. 2 weeks after Tx all animals were normoglycemic. Over the following 6 months nearly normal plasma glucose levels could be maintained in the KTx group whereas the STx animals already became diabetic after 3 months. An oral glucose load after 2 months showed slightly elevated plasma glucose levels in the KTx group which only gained statistical significance in a similar test after 6 months. In the STx group definitely impaired glucose tolerance already prevailed at 2 months. An i.v. glucose tolerance test performed 6 months after Tx showed a loss of first phase insulin release in both Tx groups but an increased second phase release and a preserved response to L-arginine compared to controls. Insulin content remained unchanged in the KTx group, but was markedly reduced in the STx group after 6 months. In conclusion we may say that KTx is able to establish near-normoglycemia in streptozotocin-diabetic Lewis rats which can be maintained over a long period of time despite abnormal glucose tolerance and impaired insulin secretion STx could normalize plasma glucose only up to 3 months showing early impairment of glucose and insulin regulation. If KTx has immunological or local advantages (islet vascularization, innervation) compared to STx with higher dispersion of islets in spleen and portal vascular system remains unclear.

Orthotopic cardiac transplantation is an excellent palliation for endstage cardiac failure in select patients. Individual patients may benefit from heterotopic cardiac transplantation. Since 1986 88 patients received an orthotopic or heterotopic transplant at Charité University Hospital. 83% one year-survival and 58% nine years-survival rates were achieved. Infection is a leading cause of death in patients after cardiac transplantation. Due to persistent scarcity of donor organs we have to consider alternative surgical methods as there are dynamic cardiomyopathy, and the chronic conventional surgery, and the chronic use of mechanical circulatory assist systems.

The ability to replace damaged myocardial tissue with new striated muscle would constitute a major advance in the treatment of diseases that irreversibly injure cardiac muscle cells. The creation of focal grafts of skeletal muscle has been reported following the intramural injection of skeletal myoblasts into both normal and injured myocardium. The goals of this study were to determine whether skeletal myoblast-derived cells can be engrafted into the murine heart following arterial delivery. The murine heart was seeded with genetically labeled C2C12 myoblasts introduced into the arterial circulation of the heart via a transventricular injection. A transventricular injection provided access to the coronary and systemic circulations. Implanted cells were characterized using histochemical staining for beta-galactosidase, immunofluorescent staining for muscle-specific antigens, and electron microscopy. Initially the injected cells were observed entrapped in myocardial capillaries. One week after injection myoblasts were present in the myocardial interstitium and were largely absent from the myocardial capillary bed. Implanted cells underwent myogenic development, characterized by the expression of a fast-twitch skeletal muscle sarcoendoplasmic reticulum calcium ATPase (SERCA1) and formation of myofilaments. Four months following injection myoblast-derived cells began to express a slow-twitch/cardiac protein, phospholamban, that is normally not expressed by C2C12 cells in vitro. Most surprisingly, regions of close apposition between LacZ labeled cells and native cardiomyocytes contained structures that resembled desmosomes, fascia adherens junctions, and gap junctions. The cardiac gap junction protein, connexin43, was localized to some of the interfaces between implanted cells and cardiomyocytes. Collectively, these findings suggest that arterially delivered myoblasts can be engrafted into the heart, and that prolonged residence in the myocardium may alter the phenotype of these skeletal muscle-derived cells. Further studies are necessary to determine whether arterial delivery of skeletal myoblasts can be developed as treatment for myocardial dysfunction.

Background: We tested the hypothesis that the University of Wisconsin solution has a ionic composition (i.e., intracellular, calcium-free, lactobionate-enriched) that may be beneficial for cold heart graft preservation independently from any additives., Methods: St. Thomas' Hospital and University of Wisconsin solutions were compared with the following: (1) C solution, a simplified University of Wisconsin-like solution (i.e., intracellular, calcium-free, lactobionate-enriched); (2) A solution, an St. Thomas' Hospital-like solution (extracellular, calcium [Ca2+] = 1.2 mmol/L) in which chloride was replaced by lactobionate; (3) B solution, an intracellular, lactobionate-enriched, calcium-containing solution ([Ca2+] = 1.2 mmol/L). Rabbit hearts were transplanted heterotopically in the abdomen of recipient animals either immediately or after 6 hours of storage. Hemodynamic parameters were recorded 60 minutes after unclamping., Results: After a 6-hour storage, University of Wisconsin and C solutions provided better preservation than B and St. Thomas' Hospital solutions: diastolic pressures were lower; developed pressure and rate of pressure rise were higher. C solution was superior to University of Wisconsin solution only for rate of pressure rise. A solution was intermediary. A significant alteration of resting pressure and hemodynamic parameters was generally observed during the 6-hour storage. Nonsignificant changes of developed pressure and rate of pressure rise were only observed in C and B solutions: This is explained by systolic alteration after immediate reimplantation for the B group and good preservation for the C group. Resting pressure was unchanged over a 6-hour storage only for the C group, but this measure was not determined for University of Wisconsin. A correlation exists for various left ventricular volumes between resting pressure and postreperfusion hemodynamic data. Replacement of chloride by lactobionate (A versus St. Thomas' Hospital) may have improved resting and diastolic pressures by other mechanisms than limitation of net water gain during storage.

Recovery of digital nerve function in 21 patients with toe-to-digit transplantation was evaluated by clinical sensory tests and somatosensory evoked potentials (SEPs) to median and digital nerve stimulation. The mean interval between injury and surgery was 7 months, and that between surgery and study was 31 months. The transplanted toes achieved a satisfactory but incomplete recovery in temperature (warm and cold), pinprick, touch, vibration, and two-point discrimination in that order. The overall sensory status of the transplanted toes appeared to be closer to normal toes than to normal fingers. In SEPs from the transplanted side, median N9, N13, and N20 components had normal latency but reduced amplitude, whereas digital N9 component was usually absent, but N13 and N20 components had prolonged latency and reduced amplitude. Transplantation performed within 1 month after injury prevented amplitude reduction in median SEPs and latency prolongation in digital SEPs. The SEP data suggest that timing of surgery was critical in preventing retrograde effect on the median nerve, and that recovery of digital nerve function was incomplete correlating with clinical sensory findings.