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Background Context: Patients with infective spondylodiscitis who failed conservative treatment are generally indicated for open surgery. However, some patients are poor candidates for standard surgery, hence the need to evaluate less invasive approaches. Good outcomes were previously reported for percutaneous suction aspiration and drainage (PSAD) in the treatment of infective spondylodiscitis resistant to conservative therapy. We recently extended the surgical approach of PSAD to allow drainage of paravertebral or epidural abscesses in patients with progressive infective spondylodiscitis., Purpose: To evaluate the clinical outcomes of PSAD for infective spondylodiscitis with paravertebral or epidural abscess., Design: Retrospective case series., Patient Sample: Patients with infective spondylodiscitis and associated epidural or paravertebral abscess treated using PSAD at our institution, between 1998 and 2014., Outcome Measures: Serum levels of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and imaging data obtained via plain radiography, computed tomography, and magnetic resonance imaging were analyzed. Serum measurements were taken preoperatively and at several time points postoperatively. Clinical outcomes were evaluated using the modified MacNab criteria for overall functional mobility., Methods: Data were obtained from the patients' case notes, radiological images, and medical records. Student t test was used to assess the relevance of changes in serum levels of CRP and ESR at each evaluated time point, as well as the change in sagittal Cobb angle between the preoperative state and the state at final follow-up., Results: Fifty-two patients (31 men and 21 women; average age, 70.6 years) were included in our analysis. The median (range) CRP levels and ESR values at the time of diagnosis were 6.86 (0.04-20.15) mg/dL and 78.8 (26-120) mm/h, respectively. At 1 year postoperatively, these values had decreased to 0.18 (0.0-1.2) mg/dL and 13.8 (4-28) mm/h for CRP and ESR, respectively. At final follow-up, bone union was observed in 80.8% (42 of 52) of patients, with instability identified in five patients. Regarding functional mobility, excellent outcomes were obtained in 26.9% (14 of 52) of patients, whereas good, fair, and poor outcomes were noted in 42.3% (22 of 52), 3.9% (2 of 52), and 26.9% (14 of 52) of patients, respectively. Overall, treatment was considered effective in 69.2% (36 of 52) of patients., Conclusions: Percutaneous suction aspiration and drainage can serve as an effective alternative to open surgery for the treatment of patients with progressive infective spondylodiscitis and associated paravertebral or epidural abscess., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: Calcineurin-inhibitor-induced pain syndrome (CIPS) was used as a reference in the literature as reflex sympathetic dystrophy syndrome related to calcineurin inhibitors. Much of the literature describes CIPS that occurred after kidney and bone marrow transplantation. We describe a rare case of CIPS in induction immunosuppression before kidney transplantation, under administration of an anti-rheumatoid drug., Methods: A 53-year-old woman had pre-status of ABO-incompatible living kidney transplantation. The patient had rheumatoid arthritis, but that was well-controlled with salazosulfapyridine as an anti-rheumatoid drug. Fourteen days before transplantation, she received induction immunosuppressive therapy consisting of tacrolimus (TAC) and mycophenolate mofetil (MMF) and she stopped taking salazosulfapyridine. The third day after that treatment, she had a high fever, fatigue, and joint pains of the knees, elbows, and wrists., Results: When the patient stopped taking TAC and MMF and started taking salazosulfapyridine again, she soon recovered. Next, we challenged same induction immunosuppression therapy with administration of salazosulfapyridine; however, the patient had the same symptom. We considered that the symptom was caused by TAC or MMF, and we did not challenge-test each drug. We found that taking only TAC caused the same symptom for the patient. Also, we challenged cyclosporine (CsA) with MMF and confirmed that she did not have the symptom., Conclusions: We decided that drugs of the induction immunosuppression therapy were CsA, MMF, prednisolone, and basiliximab. The patient received induction therapy with plasmapheresis and rituximab in addition to the above-mentioned drugs, and we performed ABO-incompatible kidney transplantation for her. The post-surgical course was good, without acute rejection, and she had no pain., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Background: Urinary decoy cells develop after renal transplantation and their appearance is attributable primarily to the proliferation of polyomavirus types BK and JC. We measured the levels of these 2 viruses that cause decoy cells to appear in the urine., Patients and Methods: BK and JC virus levels were quantified in 1182 urine samples from 335 renal transplant patients using a multiplex Taqman real-time polymerase chain reaction assay. Forty-four samples were excluded from analyses because both viruses were present at ≥10(4) copies/mL. We analyzed the relationship between viral load and the presence of urinary decoy cells., Results: Decoy cells were observed in 237 of 1138 urine samples (21%) and the BK and JC viruses were positive in 205 (18%) and 455 (40%) samples, respectively. Decoy cells were observed in 0%, 21%, 67%, 87%, 100%, and 96% of urine samples when the BK viral load was <10(4), 10(4)-10(5), 10(5)-10(6), 10(6)-10(7), 10(7)-10(8), and ≥10(8) copies/mL, respectively; and in 1%, 13%, 41%, 59%, 87%, and 97% of urine samples when the JC viral load was <10(4), 10(4)-10(5), 10(5)-10(6), 10(6)-10(7), 10(7)-10(8), and ≥10(8) copies/mL, respectively., Conclusions: BK virus more frequently triggered the appearance of decoy cells than did JC virus at equivalent viral titers., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Background: Pancreatic islet transplantation has emerged as an effective treatment for type 1 diabetes mellitus, but its use is limited due to an insufficient supply of cadaveric pancreata. In Japan, uncontrolled donors after cardiac death (DCD) are not deemed to be suitable for whole-organ pancreatic transplantation, and can provide a source of pancreas for islet transplantation. However, the long-term outcomes and utility of uncontrolled DCD in the clinical setting remain controversial. Here, we summarize the long-term outcomes of islet transplantation employing uncontrolled DCD as reported to the Japan Islet Transplantation Registry., Methods: Sixty-four isolations and 34 transplantations of pancreatic islets were conducted in 18 subjects with type 1 diabetes mellitus under the cover of immunosuppression with basiliximab, sirolimus, and tacrolimus. All donors were uncontrolled DCD at the time of harvesting. The mean follow-up time was 76 months., Results: Of the 18 recipients, 8, 4, and 6 recipients received 1, 2, and 3 islet infusions, respectively. Overall graft survivals (defined as a C-peptide level ≥0.3 ng/mL) were 72.2%, 44.4%, and 22.2% at 1, 2, and 5 years, respectively, whereas the corresponding graft survivals after multiple infusions were 90.0%, 70.0%, and 30.0%, respectively. Three of these recipients achieved insulin independence in 14, 79, and 215 days. HbA1c levels and the requirement of exogenous insulin were improved before loss of graft function. All recipients became free of severe hypoglycemia unawareness, however, at least 5 of 14 patients who had graft failure experienced recurrence of severe hypoglycemia after the loss of graft function., Conclusions: Islet transplantation from DCD can relieve glucose instability and problems with hypoglycemia when the graft is functioning. However, islets from uncontrolled DCD may be associated with reduced long-term graft survival. Further improvements in the clinical outcome by modification of islet isolation/transplantation protocols are necessary to establish islet transplantation using DCD., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Background: Quantification of the serum level of BK virus is used as a surrogate marker for the early onset of BK virus nephropathy. However, little is known about the diagnostic value of the urine level of BK virus for nephropathy or the relationship between the serum and urine viral load. We investigated the correlation between urine and serum BK virus levels after renal transplantation., Methods: From November 2008 to August 2013, a total of 270 renal transplant patients who were followed at our institution were included in this study. Urine and serum were collected simultaneously. BK virus levels were quantified in 894 urine and serum samples using a real-time polymerase chain reaction assay., Results: BK virus was detected in 178 urine samples and 36 serum samples. Among the BK virus-positive urine subjects, the positive predictive value for viral detection in the serum was 9% (13/147) when the urinary virus level was <10(7) copies/mL and 74% (23/31) when the urinary virus was ≥ 10(7) copies/mL. Serum BK viral levels were ∼2-3 log units lower than those in urine., Conclusions: BK virus was detected more frequently in serum when present in urine at ≥ 10(7) copies/mL after renal transplantation., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Background: Little is known about the timing of polyomavirus reactivation and its presence in urine after renal transplantation. The purpose of this study was to investigate the prevalence of positive polyomavirus in urine at various time points after renal transplantation., Methods: From November 2008 to August 2013, 279 renal transplant patients from our institution were included in this study. One urine sample was collected at 0-3, 4-6, 7-12, 13-24, 25-60, and ≥ 61 months after renal transplantation. A total of 394 urine samples were assessed for the presence of the BK and JC viruses with the use of a real-time polymerase chain reaction assay., Results: BK virus was detected in the urine of one-third of patients during the first 6 months. Thereafter, the positivity rate decreased gradually to 12% >5 years after transplantation. The positivity rate for the JC virus in urine was 33%-49% regardless of the post-transplantation phase., Conclusions: BK virus was detected more frequently in urine during the early phase after renal transplantation, whereas the JC virus was detected more consistently., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Background: We established a procedure to engineer therapeutic neo-islets in subcutaneous spaces in mice by transplanting contiguous layers of islet cell sheets. In this study, we investigated the cellular arrangements of α and β within these engineered neo-islets in vivo as a function of time after sheet transplantation., Methods and Results: Temperature-responsive culture dishes optimized for dispersed islet cell culture were prepared by covalently immobilizing a temperature-responsive polymer poly(N-isopropylacrylamide) (PIPAAm) on plastic dishes followed by laminin-5 coating. Dispersed islet cells obtained from Lewis rats were plated onto the PIPAAm dishes. After reaching confluence at day 2, islet cells were harvested as uniformly spread islet cell sheets by lowering the culture temperature from 37°C to 20°C for 20 minutes. Islet sheet transplantation was performed to creat neo-islet tissues in the subcutaneous spaces of SCID mice with streptozotocin-induced diabetes. This neo-islet engineering approach successfully lowered mouse blood glucose levels, achieving euglycemia at day 5 and thereafter. Histologic analyses of samples obtained at day 4 revealed that neo-islet tissues in the subcutaneous spaces showed heterogeneous cellular alignment of α and β cells. In contrast, analyses of samples at days 14 and 60 revealed α and β cells predominantly located at the peripheral and central parts of the engineered tissues, respectively., Conclusions: Reassembly of α and β cells occurred in neo-islet tissues engineered by sheet transplantation. The unique cellular arrangements in neo-islet tissues, which were similar to those in naïve pancreatic islets, may contribute to their longevity and long-term function., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Neither pregnancy nor birth is easy in female patients with chronic renal failure, but after kidney transplantation, childbirth is possible when the graft function is good. There are few guidelines for pregnancy permission and multiple reports of decreased transplanted kidney function after pregnancy. In this study, we analyzed factors that influenced transplanted kidney function deterioration during pregnancy. Twenty-one women among 33 total pregnancies have given birth in our institution. Factors analyzed were donor and recipient age at transplantation, birth age of recipient, living or cadaveric donor, hemodialysis period before transplantation, delivery method, presence of hypertension and protein urea at the beginning of pregnancy, and period between pregnancy and transplantation. Maternal graft function at the beginning of the pregnancy was 1.16 ± 0.39 mg/dL (range = 0.5-2.1). A rise in serum creatinine (S-Cr) before delivery was observed in 10/21 cases: six cases showed a rise in S-Cr levels at 1 or more years after delivery. From the analysis, graft function at the beginning of pregnancy became a significant factor correlating with the elevation of S-Cr levels during pregnancy (P = .002). Patients were divided into two groups by S-Cr levels at the beginning of pregnancy: group A was S-Cr ≤ 1; group B was S-Cr 1-2 mg/dL. All group A cases showed stable graft function before and after delivery. Some individuals in group B experienced deterioration of graft function during pregnancy; the others had stable graft function. The presence of treated hypertension at the beginning of pregnancy in group B significantly impacted renal dysfunction during pregnancy (P < .05). In conclusion, the presence of treated hypertension at the beginning of pregnancy was a significant risk factor for functional deterioration of the transplanted kidney during pregnancy even if the individual was initially within pregnancy permission criteria., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Capillary C4d was an established marker of antibody-mediated rejection in kidney transplantation. Recently, C4d fixation to antibodies against HLA-coated beads were found to be a novel way to test for complement-fixing antibodies in the serum. To find differences between complement-fixing and nonfixing antibody assays, we tested HLA antibodies and C4d in 102 sera of graft-failure patients using the LABScreen and FlowPRA bead methods. HLA antibodies were observed in the sera of patients who subsequently failed and who continued to function. C4d fixing antibodies were present in the sera of patients who subsequently experienced graft failures and those with high fluorescence intensity of HLA antibodies by FlowPRA. In antibody dilution studies with LABScreen, we noted that C4d fixation occurred only at high antibody titers, and fluorescence was lost as sera were diluted. In conclusion, the fluorescence levels of antibodies were more important that the C4d content. Quantitative aspects of antibody testing using the fluorescence value appears to be a valuable factor when examining anti-HLA antibodies posttransplantation., (Copyright © 2010 Elsevier Inc. All rights reserved.)

The worldwide shortage of deceased-donor kidneys for transplantation has become a serious issue in the past decade, leading to study of marginal donors. However, both the availability and the utility of kidneys from deceased donors are still unclear. The aim of the present study was to evaluate another method to estimate donor kidney function rather than using donor creatinine (Cr). We studied 129 recipients of deceased-donor kidneys from Maastriche donor categories III and IV. We analyzed donor Cr levels before death and recipient Cr levels at 1 year after transplant, as well as estimated glomerular filtration rates (eGFR). There was no significant difference in donor Cr levels at admission to the hospital and before death according to eGFR at 1 year after transplantation: <30 mL/min/1.73 m(2) versus ≧30 mL/min/1.73 m(2). However, recipients whose donors showed lower average eGFR levels on admission displayed better renal function at 1 year after transplant (P = .025). In conclusion, donor Cr levels before death was a less useful measurement to relate to recipient renal function; eGFR provided a better index., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Purpose: The potential for introducing transmissible spongiform encephalopathy (TSE) into islet cells was indicated by recognizing that Liberase HI is isolated from Clostridium histolyticum grown in media containing brain-heart infusion broth. A national team within the Japanese Pancreas and Islet Transplantation Association implemented an islet transplantation program in Japan using Liberase HI. The program comprised 65 islet isolations from non-heart-beating donors and 34 transplants into 18 patients. Herein, we have summarized how the Association followed these recipients over the long term., Procedures: We established an ad hoc committee to follow recipients transplanted with islets isolated using Liberase HI after becoming informed of the associated dangers of using this enzyme. We also stopped islet transplantations using Liberase. The committee addressed the major concerns of the risk of the collagenase being contaminated with TSE and of the recipient follow-up. All recipients were examined by diffusion MRI and EEG and then scheduled for evaluation and follow-up by specialists in Creutzfeldt-Jakob disease (CJD). Bioassays of bovine spongiform encephalopathy prions in the enzyme proceeded using knock-in mice expressing bovine prion protein. These assays could detect contaminating prions at a dilution of 1 × 10(4). After inactivating its collagenase activity, Liberase HI was injected into the abdominal cavities of knock-in mice. Four months later, prion infectivity in Liberase HI was evaluated by immunohistochemical staining and Western blotting of spleen homogenates using anti-prion protein antibodies., Main Findings: Western blotting and immunohistochemical staining did not detect prions in Liberase HI. Diffusion MRI and EEG evaluations performed by CJD specialists confirmed that none of the transplanted recipients had CJD., Conclusions: Three years of follow-up revealed that none of the Japanese recipients of islet transplants developed CJD. Prion bioassays showed that the Liberase HI used to isolate islets for transplantation was free of infectious TSE prions., (Copyright © 2010 Elsevier Inc. All rights reserved.)

It is known that administration of mycophenolate mofetile (MMF) is associated with BK virus (BKV) nephropathy in renal transplant recipients. To determine any inhibitory effect of mizoribine for BKV, seven patients with positive BKV in their urine who took MMF as immunosuppressive therapy were evaluated after MMF was changed to mizoribine. Baseline BKV DNA in urine, which ranged from 2.2 x 10(2) to 5.5 x 10(6) copies per milliliter, decreased in all cases (mean = 1.9 x 10(-1) times; median 2.8 x 10(-3) times). Four cases turned negative within 6 months and one within 12 months. No acute rejection or deterioration of graft function occurred during the administration of mizoribine. An inhibitory effect of mizoribine on BKV was suggested.

In Japan, organ donation has been still limited because of the strict donor criteria. The aim of this study was to show the effectiveness of pancreas transplantation (PTx) by analyzing the outcomes even under poor donor conditions. Thirty-six cases of PTx (32 simultaneous pancreas and kidney transplantations [SPK], 4 pancreas after kidney transplantations) performed during the last 8 years were examined especially for donor characteristics. Mean donor age of 41.4 +/- 11.9 years was considerably older compared with that in the United States and Europe; donors aged over 40 years comprised 67% of the total. According to the criteria described by Kapur, 29 cases (81%) in our series would be considered marginal. Thus, to increase blood supply into the pancreatic head, the gastroduodenal artery (GDA) was anastomosed using donor artery to common hepatic artery or iliac Y graft. These procedures were performed in 16 of the 24 cases in which there was liver procurement. Eventually, 34 cases (94%) preserved GDA continuity. Mean total cold ischemic time of pancreatic grafts was 12 hours 15 minutes. Of 214 registrants, 17 patients on the waiting list for SPK died of diabetic complications. To date, patient survival remains 100% with a mean follow-up period of 33 months. Pancreas graft survivals at 1, 3, and 5 years posttransplantation were 92%, 80%, and 80%, respectively. In contrast, kidney survivals were 91%, 91%, and 91%, respectively. The integrity of the pancreas head and duodenum by preservation of the GDA continuity might have decreased the risk associated with the marginal donors.

The role of bone marrow (BM)-derived cells in the process of pancreatic islet regeneration remains unclear. The purpose of this study was to determine the role of BM cells in the repair process or regeneration of pancreatic islets in mice using chimeric green fluorescent protein (GFP) expressing BM cells. BM-infused chimeric mice were made diabetic by streptozotocin (STZ) injection or 60% partial pancreatectomy. GFP-positive cells within the islets and pancreas were studied immunohistologically. STZ treatment induced a 10-fold increase in PCNA-positive cells within the islets on day 7 posttreatment. GFP-positive cells increased in number within the islets as well as in the pancreatic parenchyma immediately after STZ injection. The partial pancreatectomy induced 2- to 3-fold increases on day 7 to 28 posttreatment. GFP-positive cells increased in number in pancreatic parenchyma but not within the islets. BM traffic to the pancreas significantly increased in the 2 models inducing islet regeneration. In both models, GFP-positive cells were not positive for antibodies against insulin, glucagon, or somatostatin, but were positive for markers of macrophages or fibroblasts, suggesting their involvement in the initiation of islet regeneration.

Dissection of the external iliac artery is a rare but serious complication after transplantation, leading to graft loss. Stent angioplasty has been accepted for iliac artery dissection, but the safety and benefits of this procedure in transplantation are unclear. We report a case of right external iliac artery dissection with simultaneous kidney and pancreas transplantation, which was noticed during the operation. A self-expandable metallic stent was inserted from the right femoral artery to the dissected area and arterial blood flow was immediately restored. The postoperative clinical course was uneventful and soon after transplantation both the kidney and pancreas graft functioned well. Together with a previously reported six cases of stent replacement with transplantation and our case, no complication related to this procedure was reported, and in six cases the arterial flow was restored and graft function was improved. Endovascular stent replacement may be a safe and effective procedure for iliac artery dissection in transplantation.

In Japan, pancreas donation had become possible from cadaveric donor sources, both heart-beating or non-heart-beating (NHB). Pancreas allografts have been distributed in the organ allocation system of the Japan Organ Transplant Network. Meanwhile, islet transplantation has been categorized as a tissue transplantation; it is free from legal restraints. Thus, pancreata for islet isolation must be obtained from NHB donors. Herein we report the starting program and preliminary results of islet transplantation in Japan. Selection and listing criteria for transplantation include regional priority, ABO blood type, previous islet transplant status with insulin independence, and a longer waiting time. Five institutes in Japan (Fukushima, Chiba, Kyoto, Kobe, and Fukuoka) are prepared to start programs. A two-layer cold storage method using perfluorocarbons and UW solution is recommended for pancreas preservation. Islet isolation and purification procedures are performed according to institute-specific protocol. Immunosuppression is based on sirolimus/tacrolimus combined with basiliximab induction. Two or three consecutive infusions of >5000 IE/kg are planned for each recipient until achieving insulin independence. Twenty-seven isolations and 14 transplants were performed in eight non-insulin-dependent diabetes mellitus (IDDM) recipients. Almost all (26 of 27) were NHB donors. All recipients are free from hypoglycemic episode after transplantation. One of these recipients is insulin independent; the others are currently on minimal doses of exogenous insulin. The feasibility of islet transplantation using NHB donors was confirmed using a two-layer cold storage method and a steroid-free immunosuppressive protocol, with a high rate of graft function.

ATP content or energy charge (EC) of islets may be a good parameter to assess viability. In this study, we examined adenine nucleotides and EC of freshly isolated or 24-hour cultured rat islets of various diameters using a novel bioluminescent enzymatic cycling assay system. For freshly isolated islets, ATP content and islet diameter showed a high correlation (r = 0.842, P < .001), but a significant correlation was not observed for cultured islets (r = 0.284) when all islets were included for the analysis. When only the cultured islets with a diameter <350 microm were included for analysis, a significant correlation was observed (r = 0.719). EC of freshly isolated islets fluctuated widely irrespective of diameter, in contrast with results of 24-hour cultured islets, which showed a stable, high EC, regardless of diameter. These data suggest that the ATP content of islets correlates with the islet size and that EC of islets widely fluctuates following isolation, indicating a significant role of monitoring ATP and EC of islets before transplantation.

Mitomycin-C (MMC) is categorized as an agent that causes genotoxic stress by triggering various intracellular signaling pathways. We have previously shown that MMC pretreatment of highly immunogenic crude islets leads to significant prolongation of graft survival in a rat-to-mouse model. In the present study, we examined whether TH1/TH2 cytokine, including the inflammatory cytokines interferon-gamma and interleukin (IL)-2, or the Th2 group, IL-4, IL-10, TNF-alpha, IL-1 beta, IL-6, GM-CSF, and transforming growth factor (TGF)-beta were up-regulated or down-regulated following MMC treatment of islets. We found changes in TGF-beta messenger RNA (mRNA) transcription as the only events among the measured cytokines. TGF-beta concentration was elevated in blebs formed under the kidney capsule, but not in the serum or ascites among animals given MMC-treated islets than in animals given untreated islets, suggesting local processes induced by MMC might inhibit xenograft rejection.

The Pancreas Transplantation (tx) Program under the Japanese Organ Transplant Act was started in 2000. PTx is indicated for type 1 diabetic patients on hemodialysis therapy. As of April 2003 93 patients are listed as candidates. Ten cases of PTx with enteric or bladder-drained technique were performed during the last 3 years as well as nine SPK and one PAK. Of 10 cases, nine recipients are insulin-free with HbA1c values ranging from 4.4% to 5.7%, although exogenous insulin was required in six cases temporarily, namely for a median 63 days (12 to 225 days). One case was lost due to pancreatic graft thrombosis. All 10 kidney grafts are functioning. Based on the experiences with 14 cases of pancreas tx using non-heart-beating (NHB) donors, we defined the criteria of NHB donor as: age younger than 40 years and cessation of respiratory support. One case of SPK with graft of NHB donor was done, and the recipient is off insulin. Pancreas and kidney are allocated for SPK if the recipient shares at least one HLA-DR antigen. Marginal donors were defined as higher mean donor age, median 37 (range 18 to 58 years); mean 38 +/- 12 years), and no death cause of by trauma. The revascularization of gastroduodenal artery to the pancreatic graft was performed in eight cases to minimize the risk of ischemic injury to the pancreatic graft and technical failure in cases of marginal donor.

Assessment of islet viability before transplantation is mandatory for successful clinical transplantation. ATP content or energy charge (EC) of islets may represent a good parameter to assess viability. We have introduced a novel bioluminescent enzymatic cycling assay using synthetic firefly luciferase, pyruvate kinase) (PK), and pyruvate orthophosphate dikinase (PPDK) to determine adenine nucleotides (AN) in isolated islets. A complete assay requires several minutes. The ATP contents of 1, 3, 10, 30 or 100 islets each with a diameter of 150 microm were 9.95 +/- 0.03, 28.3 +/- 1.18, 89.5 +/- 1.48, 282 +/- 10.2, and 673 +/- 27.1 pmol, respectively, showing a relatively constant ATP content per 30 islets (9.95, 9.42, 8.95, and 9.41 pmol/IEQ). ECs of each group were 0.74 +/- 0.02, 0.74 +/- 0.02, 0.75 +/- 0.02, 0.74 +/- 0.02, and 0.65 +/- 0.01, respectively, a value that was quite constant up to 30 islets. The quantity of ATP and EC in a single islet can be measured quickly and reproducibly, offering a new method to determine viability of isolated islets prior to transplantation.