Kazunobu Shinoda

Regulation of human dendritic cells by a novel specific nuclear factor-κB inhibitor, dehydroxymethylepoxyquinomicin

Regulation of antigen-presenting cells (APC) is crucial in controlling allograft rejection. Dendritic cells (DC) are the most potent APC and must mature to present antigens to T-cell receptors. During DC maturation, nuclear factor-kappaB (NF-kappaB) is a key transcriptional factor. We synthesized dehydroxymethylepoxyquinomicin (DHMEQ), which specifically inhibits the final step of nuclear translocation of activated NF-kappaB proteins and examined its immunoregulatory effects on human monocyte-derived DC (Mo-DC). Regulatory Mo-DC were generated by pretreatment with DHMEQ before LPS stimulation, which were termed dl-DC. DHMEQ pretreatment (5 microg/ml) completely inhibited nuclear translocation of activated NF-kappaB. DHMEQ significantly inhibited DC production of proinflammatory cytokines (IL-6, TNF-alpha, and IL-12 p70) in a dose-dependent manner. IL-12 was most potently inhibited. However, IL-10 production by dl-DC was only moderately affected by DHMEQ. Although CD40 and the expression of HLA-DR (HLA-DR) expression on dl-DC was downregulated, CD80 and CD86 expression was moderately upregulated. Induction of T helper 1 cell responses was efficiently impaired by dl-DC. This confirmed that DHMEQ-treated Mo-DC exhibited immunoregulatory effects. These findings suggest that DHMEQ has potential as an immunosuppressive drug for human immune cells.

Effect of a novel nuclear factor-κB activation inhibitor on renal ischemia-reperfusion injury.

Background In kidney transplantation, the relationship between prolonged warm or cold ischemic storage of kidneys and a higher incidence of delayed graft function is previously known, and delayed graft function has been known to aggravate poor long-term graft survival. We investigated the effect of a novel nuclear factor-&kgr;B activation inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), on renal ischemia-reperfusion (I/R) injury. Methods DHMEQ was administered to Lewis rats once just before renal artery clamping (DHMEQ pretreatment group), and the effect on I/R injury was investigated. Results In the DHMEQ pretreatment group, the 24-hr urine volume on days 1 to 3 after I/R was significantly larger, and the protein concentration of the urine on days 2 to 7 was significantly smaller than in the untreated group. The serum creatinine level was significantly improved, and significantly lower levels of the inflammatory cells and inflammatory cytokines were present in the kidneys on day 1. The relative ratio of nuclear to cytoplasmic nuclear factor-&kgr;B and oxidative stress of kidney tissue on day 1 were significantly decreased. Conclusions Treatment with DHMEQ before renal artery clamping may therefore be useful for renal I/R injury and application to renal transplantation is expected.

Successful low-dose leflunomide treatment for ganciclovir-resistant cytomegalovirus infection with high-level antigenemia in a kidney transplant: A case report and literature review.

Ganciclovir-resistant cytomegalovirus infection is sometimes life-threatening for organ transplant recipients. Foscarnet is an alternative, although it may potentially worsen the preexistent impaired renal function. Here we report the case of a successful low-dose leflunomide treatment in a kidney transplant recipient with very high viral replication, who underwent kidney transplantation 10 years before. Administering 10mg leflunomide daily for 5 months without a loading dose completely cleared the ganciclovir-resistant cytomegalovirus strains.

Renal Arteriolar Injury by Salt Intake Contributes to Salt Memory for the Development of Hypertension

The role of salt intake in the development of hypertension is prominent, but its mechanism has not been fully elucidated. Our aim was to examine the effect of transient salt intake during the prehypertensive period in hypertensive model animals. Dahl salt-sensitive rats and spontaneously hypertensive rats were fed from 6 to 14 weeks with low-salt (0.12% NaCl), normal-salt (0.8% NaCl), high-salt (7% NaCl), or high-sodium/normal-chloride diet and returned to normal-salt diet for 3 months. Rats in the high-salt group saw elevations in blood pressure (BP) not only during the treatment period but also for the 3 months after returning to normal-salt diet. We named this phenomenon salt memory. Renal arteriolar injury was found in the high-salt group at the end of experiment. Dahl salt-sensitive rats were fed from 6 to 14 weeks with high-salt diet with angiotensin receptor blocker, vasodilator, calcium channel blocker, and calcium channel blocker+angiotensin receptor blocker and returned to normal-salt diet. Although BP was suppressed to control levels by vasodilator or calcium channel blocker, elevated renal angiotensin II and renal arteriolar injury were observed, and salt memory did not disappear because of sustained renal arteriolar injury. Calcium channel blocker+angiotensin receptor blocker suppressed renal arteriolar injury, resulting in the disappearance of salt memory. Cross-transplantation of kidneys from Dahl salt-sensitive rats on high salt to control rats caused increase of BP, whereas control kidneys caused reduction in BP of hypertensive rats, inducing the central role of the kidney. These results suggest that renal arteriolar injury through BP and renal angiotensin II elevation plays important roles in the development of salt memory for hypertension.

Are predictive models for cancer volume clinically useful in localized prostate cancer

Objectives:  To evaluate the correlation between preoperatively predicted and pathologically measured prostate cancer volumes and to investigate the clinical use of preoperatively predicted cancer volume in predicting pathological stage.

Should We Try Antiandrogen Withdrawal in Castration-Resistant Prostate Cancer Patients? Insights From a Retrospective Study.

BACKGROUND It remains uncertain whether those with response to antiandrogen withdrawal (AAW) have a better prognosis. We investigated the predictors of a better response to AAW and overall survival after acquiring resistance to first-line androgen deprivation therapy inpatients with castration-resistant prostate cancer (CRPC). PATIENTS AND METHODS We retrospectively reviewed the medical records of 87 CRPC patients treated at Keio University Hospital. Sixty-seven of 87 CRPC patients underwent AAW. We analyzed clinicopathologic parameters to identify predictors of survival in CRPC patients and investigated predictors of good response to AAW. RESULTS Younger age, longer duration of androgen deprivation therapy before CRPC development, and better response to AAW were independent favorable prognostic factors for overall survival. Although better response to AAW was a favorable prognostic factor in this study, trying AAW was not significantly related to overall survival. Duration of hormone therapy was significantly longer in those whose disease responded to AAW (69.9 ± 11.0 months) than those with no response (45.3 ± 5.2 months). CONCLUSION The prognostic benefit of AAW was not clearly determined in this study. However, AAW might be beneficial in patients who have favorable prognostic factors for a response to AAW-that is, those who have received hormone therapy for a long period. However, AAW should not be done in patients who do not have favorable factors and who had a high prostate-specific antigen level at the time of their prostate cancer diagnosis.

Recombinant Thrombomodulin Alpha Protects Kidney Graft from Damage after Long Cold Ischemia Time in a Rat Kidney Transplant Model

Objectives Thrombomodulin (TM) extensively expresses on the endothelial cells in the steady state and is known to prevent hypercoagulation via combining with thrombin and inactivating its procoagulant activity. TM also has anti-inflammatory effects. In inflammatory situation including ischemia reperfusion injury TM is known to decrease its expression on the endothelial cells. The purpose of this study is to investigate whether perfusate solution saturated with recombinant TM alpha (rTM) can protect the kidney grafts from the damage after long cold ischemia time. Methods We employed a rat syngeneic kidney transplant model using Lewis strain. Donor kidney grafts were preserved in a cold UW perfusate solution saturated with rTM (Group A, n=7), or not (Group B, n=8) for 24 hours. Then kidney transplantation (Lewis to Lewis) was performed. Blood and urine samples were sequentially collected (post operative day 1, 2, and 7) to measure creatinine clearance. Neutrophil gelatinase-associated lipocalin (NGAL), an acute kidney injury marker, was also measured by ELISA. Kidney graft samples in an acute phase (2 hours after operation) were collected to investigate the grade of tubular damage. TUNEL assay by immunofluorescence was performed. Immunohistochemistry (IHC) for TM was also performed. Results IHC revealed that the expression of TM on kidney grafts after 24-hour cold ischemia was extensively reduced compared with those after 0-hour ischemia. Creatinine clearance (ml/min/kg) in Group A was significantly ameliorated compared with that in Group B: mean (SD), 95% CI of Group A vs Group B; 0.68 (0.33), 0.41-0.96 vs 0.23 (0.13), 0.11-0.35 on POD 1; 1.22 (0.40), 0.86-1.59 vs 0.59 (0.30), 0.31-0.86 on POD 2; and 2.63 (1.41), 1.45-3.81 vs 1.17 (0.69), 0.47-1.87 on POD 7; p=0.02, two-way repeated measured ANOVA. Furthermore, serum levels of NGAL (mg/ml) in Group A were significantly lower than those in Group B: mean (SD), 95% CI of Group A vs Group B; 6.5 (1.7), 0.70-4.71 vs 9.1 (1.1), 7.8-10.4 on POD 1; and 4.3 (2.7), 1.5-7.1 vs 8.9 (1.8), 6.6-11.1 on POD 2; p=0.021, two-way repeated measured ANOVA. Fewer damaged tubular cells and apoptosis positive cells at a very acute phase were observed in Group A (Figure 1), compared with in Group B (Figure 2). Conclusion Perfusion with rTM significantly attenuates kidney graft damage after long cold ischemia time by abrogating apoptosis in tubular cells Figure. No caption available. Figure. No caption available.

Prognostic Value of Baseline Serum C-Reactive Protein Level in Intermediate-Risk Group Patients With Metastatic Renal-Cell Carcinoma Treated by First-Line Vascular Endothelial Growth Factor–Targeted Therapy

Micro‐Abstract Because accumulating evidence underlines the association of systemic inflammation with metastatic renal‐cell carcinoma (mRCC) progression, we evaluated baseline C‐reactive protein (CRP) levels as a prognostic marker in 107 intermediate‐risk mRCC patients treated with first‐line targeted therapy. Baseline CRP could be a biomarker correlated with overall survival in the intermediate‐risk group. Its cost efficacy and availability make CRP a helpful tool for reclassifying the intermediate‐risk group. Background: Almost half of patients with metastatic renal‐cell carcinoma (mRCC) are classified as intermediate risk by the International Metastatic Renal Cell Carcinoma Database Consortium (IMDC) model. The aim of this study was to evaluate whether baseline C‐reactive protein (CRP) levels predict overall survival (OS) in intermediate‐risk group mRCC patients. Patients and Methods: Data from 107 intermediate‐risk group mRCC patients receiving first‐line targeted therapy were retrospectively reviewed. We evaluated the correlation between baseline CRP levels as well as other indices and OS. Results: Of the 107 patients with intermediate‐risk disease, 46 patients (43%) were classified as having elevated CRP levels. The elevation of pretreatment serum CRP levels was the independent prognostic factor of OS in patients with intermediate risk (hazard ratio, 4.609; P = .001). The 1‐ and 3‐year survival rates of patients with intermediate–nonelevated CRP were 90.0% and 64.7% compared to the favorable‐risk group, at 92.1% and 68.5%, respectively. In contrast, the 1‐ and 3‐year survival rates of patients with intermediate–elevated CRP were 80.5% and 37.4% compared to the poor‐risk group, at 65.2% and 24.2%, respectively. Conclusion: Baseline CRP levels could divide mRCC patients in the intermediate‐risk group into 2 prognostic subgroups.

Change of the 5α/5β ratio of urinary steroid metabolites in benign prostatic hyperplasia patients treated with dutasteride

BACKGROUND The effects of the administration of dutasteride (DUT) on steroid metabolite pathways in BPH patients have not been examined. METHODS Urine and blood samples as well as clinical parameters were prospectively collected after the administration of DUT to 60 BPH patients, and after its withdrawal in another set of 25 BPH patients. Urine samples were assessed using gas chromatography/mass spectrometry for the urinary steroid profile (USP), which simultaneously measures 63 steroid metabolites. We examined pharmacological changes in the 5α/5β ratio of urinary metabolites and their relationships with clinical parameters in patients treated with DUT. RESULTS The mean urinary androsterone/etiocholanolone (An/Et) ratio in sex-steroid pathways significantly decreased from 1.39 to 0.02 (p < 0.01). Urinary metabolites in other steroid pathways such as 5αTHF/5βTHF in the glucocorticoid pathway and 5αTHB/5βTHB in the mineralocorticoid pathway also significant decreased after the DUT treatment. As compared to baseline level, the mean An/Et ratios in patients with the withdrawal of DUT were 0.7%, 1.4%, 12.6%, and 82.4% at just before, one month, 3 months, and 6 months after the withdrawal of DUT, respectively. All other steroid pathways changed in a similar manner without the aggravation of urinary symptoms. The recovery ratio of An/Et in USP before and 3 months after the withdrawal of DUT correlated with the recovery ratio of serum PSA levels (ρ = 0.61, p < 0.01). CONCLUSION Urinary 5α/5β metabolites in all pathways were strongly suppressed after the administration of DUT for one month and the pharmacological effect of DUT prolonged even after withdrawal of DUT.

MP74-15 A NOVEL NF-?B INHIBITOR DEHYDROXYMETHYLEPOXYQUINOMICIN PREVENTS ACUTE REJECTION OF KIDNEY ALLOGRAFTS IN A RAT MODEL

performed in 3 patients with chronic flank pain prior to surgery. Age, BMI, Creatinine, length of follow up, subjective pain score and use of narcotics at last visit were assessed. RESULTS: Of the 6 patients included in the study, 5 underwent auto-transplant. Of these 5, 2 had celiac block prior to surgery, both of which reported alleviation of pain following the block. Of note, the patient who did not undergo auto-transplant also had a successful celiac block but elected for simple nephrectomy for fear of complications with transplantation. All 3 patients who had celiac block and subsequently underwent surgery had resolution of pain and were not on narcotics at last follow-up. Of the remaining 3 patients, 2 were still taking narcotics at last follow up. Mean age was 36.8 yrs., mean BMI was 26.2 and mean follow up was 11.7 mos. All patients had normal renal function at last follow-up with mean serum Cr of 0.88 mg/dL. CONCLUSIONS: Chronic pain syndromes are frustrating for patients, their families, and their physicians. Invasive surgical options such as auto transplant or nephrectomy are considered when conservative measures fail, but not even these methods guarantee pain relief, and can result in significant complication. Based on our experience, we recommend celiac plexus block prior to consideration of more invasive treatment when conservative options fail.