Serum level of full-length connective tissue growth factor reflects liver fibrosis stage in patients with Fontan-associated liver disease.

BACKGROUND: Chronic liver disease leads to liver fibrosis, and an accurate diagnosis of the fibrosis stage is crucial for medical management. Connective tissue growth factor (CTGF) is produced by endothelial cells and platelets and plays a central role in inducing fibrosis in various organs. In the present study, we tested the validity of measuring the serum levels of two types of CTGF to estimate the biopsy-confirmed liver fibrosis stage. METHODS: We used two detection antibodies targeting the N- and C-terminal of CTGF to measure the serum levels of two forms of CTGF consisting of its full length and its N-terminal fragment. We analyzed the level of CTGF (via enzyme-linked immunosorbent assay) and the liver fibrosis stage in 38 patients with Fontan-associated liver disease (FALD) (26 cases of which were diagnosed pathologically). Correlations were determined by multivariate analysis and the area under the receiver operating characteristic curve. The 65 patients with nonalcoholic fatty liver disease (NAFLD) were included as a disease control group for examination. RESULTS: Full-length CTGF was significantly inversely correlated with liver fibrosis in patients with FALD. Although the platelet count was also associated with the liver fibrosis stage, full-length CTGF was more closely correlated with the fibrosis stage. Furthermore, the level of full-length CTGF was inversely associated with high central venous pressure. Conversely, the serum level of CTGF was not correlated with the fibrosis stage in NAFLD. CONCLUSION: The serum level of full-length CTGF may be useful for estimating the liver fibrosis stage in patients with FALD.

5-Aminolevulinic acid-based photodynamic diagnosis for detection of urothelial carcinoma cells in bladder washing sediment suspension: A pilot study.

BACKGROUND: Bladder cancer is a common malignant disease in developed countries. Early detection of malignancy is important using urine cytology. The 5-aminolevulinic acid (ALA)-based photodynamic diagnosis (ALA-PDD) has not been routinely applied in urine cytology analysis yet, although it has been well accepted for tumor lesion marking in cystoscopy. METHODS: A total of eight volunteers were enrolled in this study. The cells of sediment suspension from bladder washing fluid and random urine were stained by ALA-induced protoporphyrin IX (ALA-PpIX) and the fluorescent intensity of ALA-PpIX was analyzed by ImageJ. RESULTS: The cutoff value of fluorescent intensity was 90.260 per pixel. The proposed protocol provided an objective fluorescent intensity for evaluation. Sensitivity was 0.931 and specificity was 1.000. CONCLUSIONS: The staining procedure applied was ALA-PpIX for suspicious cells in the cellular suspension from bladder wash fluid and random urine. ImageJ was applied to the objective measurement for the fluorescent intensity of the stained cells. The cutoff value for the positive result was 90.260 per pixel. Therefore, the protocol proposed in this study provides a potential means to enhance accuracy for urine cytology analysis.

Kidney transplantation in a recipient with anti-HLA antibody IgM positive.

In general, anti-HLA antibody belongs to the IgG subclass, but there are very few reports of detection of anti-HLA IgM antibodies. In the present study, we report a renal transplant recipient with a positive NIH-complement dependent cytotoxicity (NIH-CDC) test. The patient was a 24-year-old male with focal segmental glomerulosclerosis (FSGS) as the underlying kidney disease. He had been on maintenance hemodialysis since December 2003 and finally received a living-donor allograft from his mother in October 2008. Pre-transplantation, the NIH-CDC test was positive for both B and T cells, but the flow-cytometric crossmatch test (FCXM) was negative for both cells. The result of the panel-reactive antibody assay (PRA)-single beads test using anti-IgM antibody as the second antibody demonstrated that the positive NIH-CDC test was due to the presence of anti-HLA IgM antibody against the donor-specific antigen A24. Biopsy specimens showed thrombus formation in a small number of glomeruli immediately after the transplantation, but this finding was no longer seen at three months postoperatively. We report successful renal transplantation in a case with anti-HLA IgM antibody.

Influence of serum high-molecular-weight and total adiponectin on arteriosclerosis in IgA nephropathy patients.

BACKGROUND/AIM: Adiponectin has attracted a great deal of attention because of its antiatherogenic properties. Previous studies have reported that high-molecular-weight (HMW) adiponectin may be the active form of this protein, but there have been no reports on the relationship between serum adiponectin and arteriosclerosis. The aim of our study was to determine whether HMW or total adiponectin levels are associated with arteriosclerosis in patients with IgA nephropathy. METHODS: We enrolled 72 patients aged 34.3 +/- 12.7 years, from whom interlobular arteries were obtained by renal biopsy in our hospital between 2003 and 2006 and who were confirmed to have IgA nephropathy. We assessed them in relation to age, gender, body mass index, presence of hypertension, serum total cholesterol, triglyceride, uric acid, high-sensitive C-reactive protein and HMW adiponectin and total adiponectin levels, creatinine clearance, and urinary protein excretion. The severity of arteriosclerosis was semiquantitatively evaluated and classified into the following four grades: 0 = none; 1 = mild; 2 = moderate, and 3 = severe. RESULTS: Multiple stepwise regression analysis showed associations between arteriosclerosis grade and age [standard regression coefficient (st beta) = 0.560, p < 0.001], total adiponectin (st beta = -0.218, p = 0.026), triglyceride (st beta = 0.222, p = 0.033), and presence of hypertension (yes = 1, no = 0; st beta = 0.182, p = 0.036) in the IgA nephropathy patients as a whole and associations between arteriosclerosis grade and age (st beta = 0.708, p < 0.001), HMW adiponectin (st beta = -0.321, p = 0.035), and triglyceride (st beta = 0.292, p = 0.038) in the male IgA nephropathy patients. CONCLUSIONS: Serum total adiponectin levels are an independent determinant of arteriosclerosis in IgA nephropathy patients. It was noteworthy that in males the serum HMW adiponectin levels correlated more strongly with arteriosclerosis grade than the total adiponectin levels did. Adiponectin may prevent renal arteriosclerosis.

Differences in humoral immunity between a non-rejection group and a rejection group after ABO-incompatible renal transplantation.

BACKGROUND: Renal transplantation across the blood barrier is a unique model for investigating the humoral response to different carbohydrate antigens. However, in such a renal transplantation, the characteristics of B cells as well as of the antibodies produced by B cells are less well defined. METHODS: In the present study we investigated B cell subsets (i.e., the CD5(+) B-1 and CD5- B-2 subsets) by flow cytometric analysis, and their subclasses of antibody, by ELISA, in patients who had undergone renal transplantation across the blood barrier. The subjects consisted of five recipients with good function (group 1) and five recipients with graft loss (group 2) accompanied by antibody-titer elevation after ABO-incompatible renal transplantation. RESULTS: The B-cell population analysis revealed that CD5(+) B-1 cells temporarily increased in all patients in both groups soon after transplantation, and that CD5- B-2 cells significantly increased 1 month after transplantation only in group 2. The antibody subclasses analysis showed mild elevation of immunoglobulin (Ig) G2 and IgM in group 1 as opposed to remarked elevation of IgG2, IgM and IgG1 in group 2. CONCLUSIONS: The results of this study suggested that CD5(+) B-1 cell T-independent activation usually occurs soon after ABO-incompatible renal transplantation, but that CD5- B-2 cell T-dependent activation occurs only in patients who experience graft rejection.

The mechanism responsible for accommodation after living-related kidney transplantations across the blood barrier.

The mechanism responsible for accommodation in renal transplantations across the blood barrier remains unclear. We recently encountered two patients with accommodated status after living-related kidney transplantations across the blood barrier. Both developed elevations of anti-blood-group antibodies to titers over 128x after transplantation, despite excellent renal function. We investigated the serum samples after the establishment of accommodation bound to the erythrocyte membrane of the donors or the third party with the same blood group. After the establishment of accommodation, the serum samples from both accommodated patients demonstrated a significant decrease in binding to the donors' erythrocyte membrane, but did not show any decrease in binding to the erythrocyte membrane of the third party. By contrast, serum samples from patients with graft loss after unsuccessful accommodation showed high anti-blood-type antibody activity directed towards both the donor's and the third party's erythrocytes. The result of this study suggests the difference of quality in antibodies produced by accommodated and nonaccommodated recipients.

Evaluation of flow cytometric panel reactive antibody in renal transplant recipients - examination of 238 cases of renal transplantation.

In Japan, the complement-dependent cytotoxicity (CDC-crossmatch) test and the anti-donor antibody flow cytometric assay (FCXM) are used to evaluate presensitization among transplantation candidates. We introduced the flow cytometric panel reactive antibody method (FlowPRA) at our institution, and in this paper, we compared the results of FCXM and FlowPRA. Sera of a total of 238 patients receiving the first graft were analyzed by FlowPRA retrospectively. Specimens from 125 of these patients were also analyzed by FCXM, and the results obtained using the two methods were compared. In addition, postoperative pathological findings by graft biopsy were examined in patients with PRA class 1(+) or PRA class 2(+). (i) Class 1 antibodies were detected in 36 of the 238 patients (15%), class 2 antibodies in six patients (3%), and both class 1 and class 2 antibodies in five patients (2%). (ii) Totally 125 patients analyzed by both FCXM and FlowPRA, 28 patients (22%) who tested negative by FCXM were, however, found to be positive by FlowPRA, and 16 of these 28 patients (57%) had shown evidence of humoral rejection suspected of antibody-mediated in the early postoperative stage. A large proportion of patients who tested negative by FCXM but positive by FlowPRA experienced rejection. Thus, for detecting 'high responders' in patients receiving the first graft, use of FlowPRA to detect antibodies may be superior to that of FCXM.

Mycophenolate mofetil suppresses the production of anti-blood type anitbodies after renal transplantation across the abo blood barrier: ELISA to detect humoral activity.

BACKGROUND: The introduction of novel immunosuppressive drugs has made it possible to achieve dramatic improvement in graft survival rates. In particular, the current immunosuppressive regimen including mycophenolate mofetil (MMF) has yielded excellent results including a nearly 100% 1-year graft survival rate at our institution in 2001. We used enzyme-linked immunosorbent assay (ELISA) to analyze humoral activity after ABO-mismatched renal transplantation using the MMF regimen. METHODS: The patient received an ABO-mismatched graft from a living related sibling. Preoperatively, he underwent plasma exchange (PEX) and double-filtration plasmapheresis (DFPP) several times to remove anti-blood type antibodies. Mycophenolate mofetil was used as one of the induction regimens, but a switch was made to other drugs because of persistent gastrointestinal tract discomfort. Mycophenolate mofetil was restarted, however, because of graft dysfunction caused by severe humoral rejection. Humoral activity in this patient was investigated by ELISA during the postoperative follow-up. RESULTS: Anti-blood type antibody immunoglobulin (Ig) M and IgG decreased immediately before the operation because of repeated PEX and DFPP. Both IgM and IgG were postoperatively stable and graft function was excellent. However, after switching from MMF to mizoribine (MZ), renal graft function gradually deteriorated, and the deterioration was associated with elevation of anti-blood type antibody, predominantly IgG. IgM antibody production was parallel to that of IgG, but was weaker. The elevated activity of anti-blood type antibody IgG decreased to the normal level as renal function recovered after MMF was restarted. CONCLUSIONS: Anti-blood type antibody IgG decreased after the administration of MMF after ABO-mismatched renal transplantation, and it increased after withdrawal of MMF. MMF seems to affect B-cell populations that produce anti-blood type antibodies after renal transplantation across the blood barrier.

Short-term changes in cholesterol metabolism in 40 patients with liver transplants from living related donors.

After liver transplantation, there remains a need for precise markers for evaluation of grafts. We investigated whether serum cholesterol value can serve as a marker for evaluation of the transplanted liver during follow-up. The effect of liver transplantation involving living related donors was investigated in 40 recipients in terms of lipid metabolism as measured by serum cholesterol. The relationship between cholesterol value after transplantation and liver graft weight/body weight (LW/BW) was also examined. Serum cholesterol increased at 10-20 days post-transplantation in successful cases, stabilizing at a value of more than 100 mg/dl after 4 weeks post-transplantation. In unsuccessful cases, serum cholesterol showed little increase in the 3 weeks after transplantation, and thereafter continued to decline. Cholesterol levels never reached 100 mg/dl in any of the unsuccessful transplantation cases. It took 45 days on average for the serum cholesterol to reach 100 mg/dl in recipients with less than 1% LW/BW ratio graft, but only 10 days in recipients with more than 3% LW/BW ratio graft. Patients who had partial liver transplantation from living related donors showed rapid recovery of cholesterol synthesis. However, patients with liver grafts required an extensive period before normalization of cholesterol synthesis, suggesting a need for long-term follow-up of graft recipients.