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OBJECTIVE To evaluate the efficacy and safety of onabotulinumtoxinA (botulinum toxin type A) 100 U in patients with overactive bladder and urinary incontinence. METHODS This was a phase III, randomized, double-blind, placebo-controlled trial in Japanese patients who were inadequately managed with overactive bladder medications (anticholinergics and/or β3 -adrenergic receptor agonists). Eligible patients were randomized 1:1 to receive a single dose of either onabotulinumtoxinA or placebo into the detrusor muscle (n = 124 each). The primary end-point was the change in the number of daily urinary incontinence episodes at week 12 from baseline. Secondary end-points included volume voided per micturition, other symptomatic measures (urinary urgency incontinence, micturition, urgency and nocturia) and patient-reported outcomes. RESULTS In the onabotulinumtoxinA group, there was a significantly greater decrease from baseline in the mean number of daily urinary incontinence episodes compared with the placebo group (2.16; P

Carbon 11-choline positron emission tomography/computed tomography (11C-choline PET/CT) and subsequent local therapy for oligometastatic prostate cancer have been reported to be effective, but their effectiveness in castration-resistant prostate cancer (CRPC) remains unclear. Here, we evaluated the findings of 11C-choline PET/CT in CRPC patients and the efficacy of local treatments in correspondence of the pathologic choline uptake. We collected 12 cases of CRPC patients who underwent 11C-choline PET/CT between 2014 and 2016. The outcomes assessed included age, the prostate-specific antigen (PSA) value, the findings of 11C-choline PET/CT, the subsequent treatments, the PSA response following the treatments, and the progression-free survival (PFS). Seven of 12 cases (median PSA, 3.29 ng/mL) had local prostate cancer and/or one or two metastatic lesions detected by the choline PET/CT. These localized lesions were treated with radiotherapy or lymphadenectomy. PSA decreased in all the seven cases and median PSA response was 86% (range, 23–100%). Median PFS was 8.5 months (range, 2.8–25.3 months). The other five cases (median PSA, 7.41 ng/mL) had multiple metastases and systemic therapies were continued in those cases. 11C-choline PET/CT and the correspondent local treatments may play an important role in the treatment sequence of CRPC in selected patients.

Mesenchymal stromal cells (MSCs) have been isolated from numerous sources and are potentially therapeutic against various diseases. Umbilical cord-derived MSCs (UC-MSCs) are considered superior to other tissue-derived MSCs since they have a higher proliferation rate and can be procured using less invasive surgical procedures. However, it has been recently reported that 2D culture systems, using conventional cell culture flasks, limit the mass production of MSCs for cell therapy. Therefore, the development of alternative technologies, including microcarrier-based cell culture in bioreactors, is required for the large-scale production and industrialization of MSC therapy. In this study, we aimed to optimize the culture conditions for UC-MSCs by using a good manufacturing practice (GMP)-compatible serum-free medium, developed in-house, and a small-scale (30 mL) bioreactor, which was later scaled up to 500 mL. UC-MSCs cultured in microcarrier-based bioreactors (MC-UC-MSCs) showed characteristics equivalent to those cultured statically in conventional cell culture flasks (ST-UC-MSCs), fulfilling the minimum International Society for Cellular Therapy criteria for MSCs. Additionally, we report, for the first time, the equivalent therapeutic effect of MC-UC-MSCs and ST-UC-MSCs in immunodeficient mice (graft-versus-host disease model). Lastly, we developed a semi-automated cell dispensing system, without bag-to-bag variation in the filled volume or cell concentration. In summary, our results show that the combination of our GMP-compatible serum-free and microcarrier-based culture systems is suitable for the mass production of MSCs at an industrial scale. Further improvements in this microcarrier-based cell culture system can contribute to lowering the cost of therapy and satisfying several unmet medical needs.

We herein describe a case of Sweet's syndrome (SS) in a patient being treated for pulmonary toxoplasmosis complicated with myelodysplastic syndrome (MDS). The patient's SS developed after the pulmonary toxoplasmosis improved following treatment. We searched his cytokine profiles comprehensively using a bead-based immunoassay. The results showed no elevation of interleukin (IL)-2, interferon (IFN)-γ or IL-17A, and IL-6 was only observed to have increased at the onset of SS, suggesting that the pulmonary toxoplasmosis had been well controlled and that chronic inflammation may have been the cause of SS. Pulmonary toxoplasmosis is an extremely rare occurrence. The cytokine profile can help to clarify the pathological condition of SS and MDS complicated with severely invasive infectious diseases.

The association between rectal bleeding and the received dose relative to the volume of the rectum is well established in prostate cancer patients who have undergone radiotherapy. The relative volume of the rectum is affected by the rectal anatomical volume, which depends on the definition of rectal length. Compared with the relative rectal volume, the absolute volume of the rectum may be more associated with rectal bleeding. The present study investigated the absolute volume of the rectum that may be used to predict late rectal bleeding following intensity-modulated radiotherapy (IMRT) and image-guided radiotherapy (IGRT). The cases of 82 patients of prostate cancer, who underwent IMRT and IGRT, were retrospectively evaluated by evaluating dose volume histograms. The median patient age was 73.4 years (range, 51.3–85.9 years). The median total prescribed dose was 76 Gy given in 38 fractions. The absolute and relative dose volumes of the rectum were evaluated by multivariate analysis, and the optimal dose to prevent rectal bleeding was determined. The actuarial ≥grade 1 rectal bleeding rate at 4 years was 4.5% (95% confidence interval, 1.5–13.4%) with a median observation period of 45.3 months. The absolute rectal volume (ml) treated with 60 Gy was the only significant risk factor for rectal bleeding (P

Recent studies have reported that mesenchymal stromal cells (MSCs) migrate to areas of inflammation and suppress adverse immune reactions. Bone marrow (BM)-derived MSCs have been successfully used in patients with acute graft versus host disease (GVHD), but the harvesting of BM carries certain risks for the donor. To circumvent these, we obtained MSCs from Wharton's jelly (WJ) derived from umbilical cord and investigated their potential for immunosuppression. In a mixed lymphocyte reaction (MLR), responder T cell proliferation triggered by allogeneic dendritic cells was inhibited efficiently by WJ-MSCs derived from the same donor of responder cells or those from a third party donor. These inhibitory effects were reversed in a dose-dependent manner in the presence of 1-methyl-DL-tryptophan, an inhibitor of the soluble factor indoleamine 2, 3-dioxygenase (IDO). Immunosuppression by WJ-MSCs was also attenuated by blocking cell-cell contact between WJ-MSCs and responder T cells using a Transwell chamber. Moreover, IDO gene expression was induced in both WJ- and BM-MSCs by inflammatory cytokine IFN-γ, but HLA-DR was expressed in BM-MSCs and not in WJ-MSCs upon stimulation by a relatively low concentration of IFN-γ. These results indicate that WJ-MSCs exert their immunosuppressive effects by cell-cell contact with activated T cells and in part through IDO, and suggest the need for cells rather than soluble factors secreted from MSCs to achieve immunosuppressive therapy in severe cases of GVHD.

Background: Mesenchymal stromal cells (MSC) are known to have the immunosuppressive ability and have been applied in clinic to treat acute graft-versus-host disease (GVHD), as one of severe complications after hematopoietic stem cells transplantation (HSCT) in Japan. However, MSC are activated to suppress the immune system only upon the stimulation of inflammatory cytokines and the clinical results of MSC therapies for acute GVHD are varied. It is ideal that MSC are primed to be activated and ready to suppress the immunity (=priming) before administration in vivo. Triptolide (TPL) is a diterpene triepoxide purified from a Chinese herb - Tripterygium Wilfordii Hook F (TWHF). It has been shown to possess anti-inflammatory and immunosuppressive properties in vitro. In this study, we aim to use TPL as the activator for umbilical cord-derived MSC (UC-MSC) to entry stronger immunosuppressive status. Methods: The proliferation of UC-MSC with TPL at the indicated concentrations for different time of 24, 48, 72, and 96 hours. Cell counting kit-8(CCK-8) was added in the culture medium to detect cell toxicity and the absorbance was measured using microplate reader. Flow cytometry was used to identify the MSC surface markers expression. TPL-primed UC-MSC were once replaced with fresh medium and co-culture with mixed lymphocyte reaction (MLR) consisted with mononuclear cells (MNCs) stained with CFSE and irradiated allogenic dendritic cell line (PMDC05) in RPMI 1640 medium supplemented with 10 % FBS (complete medium). IDO-1, SOD1, and TGF-β gene expression in TPL-primed UC-MSC and UC-MSC induced by 10 ng/ml IFN-γ and/or 15 ng/ml TNF-α were evaluated by RT-PCR. PDL1 and PDL2 expression in TPL-primed UC-MSC and UC-MSC in response to IFN-γ and/or TNF-α were checked by Flowjo. Results: Exposure of TPL for UC-MSC for 72hour at the concentration above 0.1 μM resulted in the cell damage significantly. Therefore, we added TPL in UC-MSC at 0.01μM of TPL for up to 48 hours, then washed thourouphly for the following culture for experiments. To evaluate the influence of TPL on the surface markers of UC-MSC, we cultured UC-MSC for 4 hours in complete medium following culture with 0.01μM of TPL for 20 hours (TPL-primed UC-MSC). TPL-primed UC-MSC revealed positive for CD105, CD73, and CD90, negative for CD45, CD34, CD14 or CD11b, CD79α or CD19 and HLA-DR surface molecules as same as the non-primed UC-MSC. In MLR suppression by UC-MSC, the TPL-primed UC-MSC activity revealed stronger anti-proliferative effect on the CD4+ and CD8+ T cells activated by allogeneic DC than those of non-primed UC-MSC in MLR. Furthermore, the TPL-primed UC-MSC promoted the expression of IDO-1, SOD1 and TGF-β in response to IFN-γ+/-TNF-α by RT-PCR and enhanced the expression of PD-L1 by FACS analysis. Discussion:In this study, we found the TPL-primed UC-MSC showed stronger antiproliferative potency on CD4+ and CD8+ T cells compared with non-primed UC-MSC. TPL-primed UC-MSC promoted the expression of IDO-1, SOD1 and TGF-β stimulated by IFN-γ+/-TNF-α, although TPL alone did not induce these factors. Furthermore, we found that the PD1 ligand (PD-L1) was induced in TPL-primed UC-MSC, likely IFN-γ enhanced the PD-L1 expression, evaluated by flowcytometry. These results suggested that TPL-primed UC-MSC seemed more sensitive to be activated as the immunosuppressant. Here, we firstly report the new function of TPL to induce the upregulation of immunosuppressive effect, although the mechanisms of TPL inhibition to MSC need to be explore. Conclusively, TPL-primed UC-MSC might be applied for the immunosuppressive inducer of MSC. Figure Disclosures He: SASAGAWA Medical Scholarship: Research Funding; IMSUT Joint Research Project: Research Funding. Nagamura:AMED: Research Funding. Tojo:AMED: Research Funding; Torii Pharmaceutical: Research Funding. Nagamura-Inoue:AMED: Research Funding.

Recently, umbilical cord (UC) has become attracted source of mesenchymal stromal cells (MSCs) for immunotherapy such as treatment of severe acute graft versus host disease, because of abundant sources and ease of collection without invasive process. In previous reports on bone marrow-derived MSCs, there is diversity of immunosuppressive mechanisms of MSCs, which have not yet fully clarified in UC-MSCs. Here we studied the immunosuppressive properties of UC-MSCs on the activated immune system in vitro. Methods; UC was collected after obtaining informed consent from the mother. UC-MSCs were obtained from frozen-thawed UC tissue with explant method and expanded in culture medium. Mixed lymphocyte reaction (MLR) consisted with mononuclear cells (MNCs) stained with CFSE and allogeneic dendritic cell (DC) line (PMDC05) co-cultured with or without UC-MSCs was carried out, and the proliferation of CD4 and CD8-positive lymphocytes were analyzed by flowcytometry followed by FlowJo software. Migratory activities of UC-MSCs toward activated lymphocytes in MLR were evaluated by counting the migrating cells stained with DAPI through the insert filter under the microscope. The percentage of CD4+CD25+FOXP3+ regulatory T cells (Treg) before and after co-culture with UC-MSCs was analyzed by flowcytometry. Monocyte phenotypes co-cultured with UC-MSCs directly or separately by insert filter were analyzed with anti-CD14, CD206, and CD168 antibodies. Results: The UC-MSCs were positive for CD105, CD73, CD90, and negative for CD45 and HLA-DR. HLA-DR, CD80, CD86, and CD40 were negative even in the high concentration of IFN-γ, while BM-MSCs became positive for HLA-DR. PD-L2 was constitutively expressed in UC-MSC, while PD-L1 was induced upon the addition of IFN-γ. In MLR, responder T cell proliferation triggered by allogeneic DC was inhibited efficiently by 3rd party derived UC-MSCs. Mean+/- SD distribution index of proliferation in CD4- and CD8- positive cells co-cultured with UC-MSCs showed 7.6+/-4.4% and 8.8+/-3.9% compared to those without UC-MSCs (n=4, as different donor derived MSCs). This inhibition was attenuated by the addition of 1-Methyl-dl-tryptophan, Indoleamine 2, 3-dioxygenase (IDO-1) inhibitor, in a dose-dependent manner. The supernatant of MLR showed the increase of IFN-γ and TNF-α, which were also suppressed in MLR co-cultured with UC-MSCs. UC-MSCs migrated toward activated lymphocytes in MLR compared with those of non-activated lymphocytes significantly, which was inhibited by the addition of AG1296, known as platelet-derived growth factor inhibitor; and PPP as insulin-like growth factor (P Discussions and Conclusion: These results demonstrated that UC-MSCs have the diverse immunosuppressive potency through migration toward the activated lymphocytes, suppressing activated T cells proliferation, regulatory T cells induction, and transition of monocyte phenotype. And UC-MSCs are the feasible and abundant source for immunotherapy. Disclosures Nagamura-Inoue: AMED: Research Funding. Nagamura:AMED: Research Funding. Tojo:AMED: Research Funding; Torii Pharmaceutical: Research Funding.

Intraventricular hemorrhage (IVH) is a frequent complication of preterm newborns, resulting in cerebral palsy and cognitive handicap as well as hypoxic ischemic encephalopathy and periventricular leukomalacia. In this study, we investigated the restorative effect on neonatal IVH by umbilical cord-derived mesenchymal stromal cells (UC-MSCs) cultured in serum-free medium (RM medium) for clinical application. UC-MSCs were cultured with αMEM medium supplemented with FBS or RM. A neonatal IVH mouse model at postnatal day 5 was generated by intraventricular injection of autologous blood, and mice were intravenously administered 1 × 105 UC-MSCs two days after IVH. Brain magnetic resonance imaging was performed at postnatal day 15, 22 and neurological behavioral measurements were performed at postnatal day 23, accompanied by histopathological analysis and cytokine bead assays in serum after IVH with or without UC-MSCs. Both UC-MSCs cultured with αMEM and RM met the criteria of MSCs and improved behavioral outcome of IVH mice. Moreover the RM group exhibited significant behavioral improvement compared to the control group. Histopathological analysis revealed UC-MSCs cultured with RM significantly attenuated periventricular reactive gliosis, hypomyelination, and periventricular cell death observed after IVH. Furthermore, human brain-derived neurotrophic factor and hepatocyte growth factor were elevated in the serum, cerebrospinal fluid and brain tissue of neonatal IVH model mice 24 h after UC-MSCs administration. These results suggest UC-MSCs attenuate neonatal IVH by protecting gliosis and apoptosis of the injured brain, and intravenous injection of UC-MSCs cultured in RM may be feasible for neonatal IVH in clinic.

Idiopathic thrombocytopenic purpura (ITP) associated with renal cell carcinoma (RCC) is relatively rare. In almost all of these case reports, patients affected with ITP developed differentially-involved cancer, because resection of cancer could not improve thrombocytopenia and the treatment of ITP needed to be continued after surgery. We report the case of a 48-year-old woman with massive renal cell carcinoma, measuring approximately 20 × 14 × 14 cm, who presented with severe thrombocytopenia: platelet count, 2000 cells/µl. After confirming normal bone-marrow, she received high-dose dexamethasone and intravenous gamma globulin, which rose the platelet count to normal levels. She then underwent left radical nephrectomy. The pathological examination revealed chromophobe RCC. After the resection, the platelet count was maintained within the normal range without any treatments. The current case is the first report of chromophobe RCC causative of severe ITP and the second case who achieved a sustained complete remission of ITP after cancer surgery alone; moreover our case is only one patient without other causes of ITP.

We experienced 4 cases of primary carcinoma of the female urethra during 1998 to 2011. All of the cases were diagnosed primary urethral cancer according to tumor biopsy, cystoscopy, and computed tomography (CT) or magnetic resonance imaging (MRI). Patients were between 66 to 92 years of age at the time of presentation. Presenting symptoms included gross hematuria in 1 case, urinary retention in another case, and vulvar bleeding in 2 cases. Pathology showed urothelial carcinoma in 2 cases, adenocarcinoma in 1 case, and squamous cell carcinoma in another case. There were 2 patients with stage D, 1 patient with C, and another with B. Three patients were treated with total cystectomy and ileal conduit. One patient was treated with radiation. Two patients died from urethral cancer, 1 patient is free from disease for 12 months, and another patient became lost during follow-up.

We report the case of a 75-year-old woman who underwent F-FDG PET/CT to evaluate the recurrence of urothelial carcinoma. PET/CT showed F-FDG-avid muscles and lymph nodes, mimicking recurrence. However, F-FDG uptake was also seen in multiple joints and subcutaneous tissue, representing an uncommon finding for recurrence. Further history taking revealed she had been pathologically diagnosed with multicentric reticulohistiocytosis (MRH) from skin biopsy, and the F-FDG PET/CT findings were consistent with MRH, a rare systemic inflammatory granulomatous disease of unknown etiology. Knowledge of the characteristic F-FDG PET/CT features of MRH is important to differentiate MRH from malignancy.

Background: For clinical cord blood (CB) transplantation, CB is processed using a standard hydroxyethyl starch protocol generally within 48 h of collection at room temperature. However, for tissue stem cell research, mononuclear cells (MNCs) were isolated from CB using a Ficoll-Paque density-gradient method. Here we report the effect of storage temperature and time from CB collection to processing on the cord blood mononuclear cells (CB-MNCs) isolated using a density-gradient method. Methods:We processed CB using a Ficoll-Paque density-gradient method to collect the cells in the MNC layer. Cells were analyzed using an automatic blood cell counter, and CD34 cells were counted according to the ISHAGEmethod. Results: The recovery rate of viable MNCs in the CB-MNC layer was inversely related to the time from collection to processing of CB samples. However, recoveries of total nucleated cell and CD34 were not affected by the time from collection to processing. The percentage of neutrophil contamination in the MNC layer increased significantly with increasing time from CB collection to processing (n=100, p

Idiopathic thrombocytopenic purpura associated with renal cell carcinoma is relatively rare. We report the case of a 48-year-old woman with massive renal cell carcinoma, measuring approximately 20 × 14 × 14 cm, who presented with severe thrombocytopenia: platelet count, 2000 cells/μL. After confirming normal bone marrow, she received high-dose dexamethasone and intravenous gamma globulin, which raised the platelet count to normal levels. She then underwent left radical nephrectomy. The pathological examination showed chromophobe renal cell carcinoma. After the resection, the platelet count was maintained within the normal range without any treatment. The current case is the first report of chromophobe renal cell carcinoma causative of severe idiopathic thrombocytopenic purpura.

Curcumin (diferuloylmethane) is a dietary phytochemical with low toxicity that exhibits growth-suppressive activity against a variety of cancer cells and possesses certain chemopreventive properties. Curcumin has already been the subject of several clinical trials for use as a treatment in human cancers. Synthetic chemical modifications of curcumin have been studied intensively in an attempt to find a molecule with similar but enhanced properties of curcumin. In this study, a series of novel curcumin analogues were synthesized and screened for anticancer activity. New analogues that exhibit growth-suppressive activity 30 times that of curcumin and other commonly used anticancer drugs were identified. Structurally, the new analogues are symmetrical 1,5-diarylpentadienone whose aromatic rings possess an alkoxy substitution at each of the positions 3 and 5. Analysis of the effects of the analogues on the expression of cancer-related genes usually affected by curcumin indicated that some induced the down-regulation of β-catenin, Ki-ras, cyclin D1, c-Myc, and ErbB-2 at as low as one eighth the concentration at which curcumin normally has an effect. The analogues, however, exhibited neither harmful nor growth-suppressive effects on normal hepatocytes where oncogene products are not activated. They also exhibited no toxicities in vivo that they may provide effective alternative therapies for the prevention and treatment of some human cancers. [Mol Cancer Ther 2006;5(10):2563–71]

BACKGROUND: Prolonged periods of marrow hypoplasia have been a problem in cord blood transplantation. DMSO is thought to produce osmotic shock to the progenitors when the thawed cells are infused into the patients. To solve this problem, a 2× dilution method originally developed in the New York Blood Center showed earlier myeloid engraftment,1 although follow-up clinical studies have not performed. STUDY DESIGN AND METHODS: To clarify the influence of the removal of DMSO by this method on the speed of engraftment in unrelated cord blood transplantation, 46 adult patients with cord blood units processed by the Tokyo Cord Blood Bank from September 1998 to March 31, 2002 were studied. Twenty-four patients received 2.6 ± 0.71 × 107 nucleated cells per kg without washing (nonwashed group), while 22 patients were received 2.7 ± 0.52 × 107 nucleated cells per kg after 2× dilution washing (washed group). RESULTS: The cumulative incidence of engraftment was not significantly different between the two groups. Median neutrophil recovery (≥5 × 109/L) in the nonwashed and washed groups was 26 and 25 days, respectively, and the median platelet recovery (≥20 × 109/L) in patients with myeloid engraftment was 44 and 40 days, respectively (NS). On the other hand, the doses of CFCs and CD34+ cells showed the influence on myeloid and platelet recovery. CONCLUSION: A 2× dilution after thawing cord blood did not result in the improvement of myeloid engraftment speed.

Quality assurance is becoming important for the cellular products in cell therapy and blood transfusion. “The International Organization for Standardization (ISO) 9002” is an International standard for quality management systems. We designed and implemented an ISO9002 system for cord blood banking. In March 2001, The Facility for Cell Processing and Preservation of the Tokyo Cord Blood Bank obtained ISO9002: 1994 from the external attestation organization as the first such of cell processing laboratory and cord blood bank in Japan.The introduction of ISO to cord blood banking ranged from reception of cord blood units, various tests, cell processing, cryopreservation, shipping and data analysis.The document system consisted of the “Quality Manual”, “General Procedure” and “Standard Operation Procedures”. The system was audited by internally and then an external attestation organization.After implementation of the ISO, several advantages were recognized including an increase in nuclear cell recovery and in the rate of medical inquiries from mothers.

Background aims Human umbilical cord (UC) has become a notable source for mesenchymal stromal cells (MSCs) that can migrate to areas of inflammation and damaged tissue and can suppress excess immune reactions and to repair, respectively. Although UC is a solid tissue, there are several advantages, including repeatable uses from the same donor sample when needed and the possibility of future explorations for cells with unknown potential, if we could cryopreserve the UC as a living tissue material. However, because the cryoprotectants in the previous reports included animal- or allogeneic human-derived serum or no serum, the frozen-thawed UC-MSCs were inferior to fresh UC-MSCs in cell proliferation. The objective of this study was to find a suitable cryopreservation method of UC for clinical use. Methods The UC was cut in cross-section and incised longitudinally, immersed in the cryoprotectant and frozen slowly. Later, it was thawed and minced rapidly, and the fragments of UC were cultured by improved explant method. Results The highest yield of cells was obtained from frozen-thawed UC with serum- and xeno-free cryoprotectant, STEM-CELLBANKER, when compared with others. The cells derived from frozen-thawed UC stored in STEM-CELLBANKER expressed the phenotypes of MSCs, retained the immunosuppressive properties in allogeneic mixed lymphocyte reactions and the differentiation potentials (into adipocyte and chondrocytes) comparable to those derived from fresh UC. Conclusions UC can be cryopreserved in serum- and xeno-free cryoprotectant as a living tissue while keeping its growth and functions equivalent to fresh UC. Our method is simple and feasible for clinical use.

The umbilical cord (UC) has become one of the major sources of mesenchymal stem cells (MSCs). The common explant method of isolating UC-derived MSCs (UC-MSCs) involves mincing the UCs into small fragments, which are then attached to a culture dish bottom from which the MSCs migrate. However, the fragments frequently float up from the bottom of the dish, thereby reducing the cell recovery rate. To overcome this problem, we demonstrate an improved explant method for UC-MSC isolation, which involves the use of a stainless steel mesh (Cellamigo(®); Tsubakimoto Chain Co.), to protect the tissue from floating after the minced fragments are aligned at regular intervals in culture dishes. The culture medium was refreshed every 3 days and the adherent cells and tissue fragments were harvested using trypsin. The number of UC-MSCs isolated from 1 g of UC using the explant method with Cellamigo was 2.9 ± 1.4 × 10(6)/g, which was significantly higher than that obtained without Cellamigo (0.66 ± 0.53 × 10(6)/g) (n = 6, p0.01) when cells reached 80-90% confluence. In addition, the processing and incubation time required to reach 80-90% confluence was reduced in the improved explant method compared with the conventional method. The UC-MSCs isolated using the improved method were positive for CD105, CD73, CD90, and HLA class I expression and negative for CD45 and HLA class II expression. The isolated UC-MSCs efficiently inhibited the responder T cells induced by allogeneic dendritic cells in a mixed lymphocyte reaction. Conclusively, we demonstrated that the use of Cellamigo improves the explant method for isolating UC-MSCs.

Examination was made of the effects of nine commercial disinfectants in short term application at concentrations recommended for practical use against 34 fungi. The effects of a yeast extract comprised of various organic substances on the efficacy of the disinfectants were also studied. Ethanol (EtOH, 70%, v/v) was found the most effective disinfectant and not influenced by yeast extract at a concentration of 5% (w/v). Oxidizing potential liquid (OPL) with 55μg/ ml as the active chlorine concentration, iodine (IA) at 45μg/ml and sodium hypochloride (NaOCI) at 200μg/ml all showed strong fungicidal effects, which were, however, significantly diminished by yeast extract. Didecyldimethylammonium chloride (DDAC) at 100μg/ml, benzalkonium chloride compounds (BACC) at 285μg/ml and benzalkonium chloride (PBKC) at 200μg/ml had a smaller fungicidal effects but there effects were less altered by the extract. [Mono-bis (trimethylammonium methylene chloride)] -alkyl (C9-15) toluene (MBAT) at 100μg/ml and chlorhexidine digluconate (CHD) at 200μg/ml, showed the smallest fungicidal effects.

We performed 82 cases of small incision radical prostatectomy from July, 2003 to September, 2009. There were 38 cases of cT1c, 41 cases of cT2, and 3 cases of cT3. Mean operative time was 222 +/- 31.7 (min.). Mean operative blood loss was 1,232 +/- 583 (ml). We evaluated factors predicting operative blood loss, such as prostate volume, body mass index (BMI), age, and preoperative PSA level. Group with smaller prostate volume and group with lower BMI showed significantly less blood loss compared to that of a higher group (p = 0.0009, p = 0.0014, respectively). Multivariate analysis showed that prostate volume and BMI were significant predictors for operative blood loss (p = 0.0005, p = 0.0122, respectively). Prostate volume and BMI may be a useful predictor for operative blood loss.

昭和63年1月より平成1年12月までの2年間に尾道総合病院皮膚科に入院した帯状疱疹患者116例について統計的観察を行った。入院患者総数に対する帯状疱疹患者の占める割合は31.0%であり, 季節的には冬と秋に比較的多かった。年齢別分布では高齢者に多く60歳代にピークを示し, 性別では女子に多かった。発症部位は顔面·頭部が最も多く, 左右差はみとめられなかった。基礎疾患を有するものは45.7%に及び, 高血圧, 癌, 肝疾患, 心疾患, 糖尿病の順に多かった。疼痛の強い期間は加齢に伴い延長し, 部位的には腹部, 上肢·胸部に発症したものに長い傾向がみとめられた。汎発化は37.1%にみとめ, より高齢者の男性に多く, 疼痛の強い期間はより遷延する傾向にあった。水痘—帯状疱疹ウイルスに対する補体結合反応抗体価は発症後2週間以内では汎発化との間に相関関係はみとめられなかったが, 発症後2週間以上を経て検索した場合には汎発例では著明に高値を示した。抗体価と疼痛の強い期間との間には, 相関関係はみとめられなかった。

The induction of hepatic drug-metabolizing enzymes by chlornitrofen (CNP) and CNP-amino was studied in the liver of male rats and mice. CNP-amino increased the activities of 7-pentoxyresorufin O -depentylase (PROD) and 7-benzyloxyresorufin O -debenzylase (BROD) as CYP2B1-dependent monooxygenase 3.6- and 4.1-fold in rats. On the contrary, these enzyme activities in mice were induced by CNP rather than by CNP-amino. Furthermore, immunoblotting showed that the protein levels of CYP2B subfamily cytoa hrome P450 (P450) in liver microsomes of rats and mice were increased by CNP or CNP-amino. Phase II drug metabolizing enzymes, UDP-glucuronyltransferase (UGT) and glutathione S -transferase (GSC) levels in mice were also significantly increased from 1.4 to 2.6-fold by CNP or CNP-amino. However, neither CNP nor CNP-amino affected UGT and GST in rats. These results suggest that CNP and or CNP-amino induce the P450 isoforms of CYP2B subfamily in the rat and mouse liver, and that the inducibility igdrugmetabolizing enzyme by the compounds is different between rats and mice.

Trichophyton verrucosum, a causative agent of bovine dermatophytosis in Japan is considered to transfer easily from infected cattle to other healthy ones. No studies have ever tried to elucidate the distribution of T. verrucosum in breeding environment. In this study, we investigated the distribution of T. verrucosum in infected young calf skin and healthy skin. Hairs or scale samples were collected from 29 lesional skin, and 46 non-lesional skin of the 19 and 34 infected calves, respectively, both varying in age 2 to 6 months old, and 35 hair samples from the 29 healthy ones. They were directly inoculated on medium. The detection rates of T. verrucosum-positive were 58.6% from the lesional parts, 34.8% from the non-lesional parts and 17.1% from the healthy parts. The isolation of T. verrucosum from the calves with no skin lesion due to dermatophytosis implied that this infection might be easily transmitted by the contact with the infected calves. T. verrucosum infection in cattle poses a serious problem in animal husbandry. It is also important for public health to take preventive measures against the infection.

Recently, cord blood transplantation (CBT) for adult is rapidly increasing in number, Especially for the patients over 50 years of age. By the end of 2003, 224 units were shipped for the patients Group I: 50 y.o.21%. We analyzed 152 patients with hematological malignancies including ALL, AML, CML, MDS, malignant lymphoma, myeloma and neuroblastoma reported from CBT center in the world by the end of 2003. Patients and Methods:Group I included 58 patients with 13 standard risk and 45 high risk patients and showed mean±SD of age; 5.3 ±4.1y.o.,BW; 20.5±13.4kg, CB volume at collection; 91.6±27.1ml, NC; 5.5±3.3x107/kg, CFC; 8.6±6.4x104/kg, CD34; 1.5±1.1x105/kg, Group II: 64 cases with 25 standard and 39 high risk patients and age; 30.6±10.3y.o., BW; 51.9±9.5kg, CB volume at collection;116.6±27.7ml, NC;2.6±0.7x107/kg, CFC;5.2±2.6x104/kg and CD34;0.8±0.5x105/kg, Group III: 30 cases with 9 standard and 21 high risk patients and age; 54.1±3.3y.o., BW;55.9±11.0kg, CB vol.at collection;118.8±24.7ml NC;2.4±0.4x107/kg, CFC;4.8±1.9x104/kg and CD34;0.8±0.4x105/kg. The patients who underwent CBT for graft failure (GF) of prior transplant were excluded. Conditioning regimen in Group I demonstrated 54 patients with full regimen and 4 with reduced intensity regimen (RIST); in Group II, 62 patients with full regimen and 2 RIST; and in Group III, 14 cases full regimen and 15 cases RIST. Results: Cumulative myeloid engraftment was seen 67.2% in Group I, 73.4% in Group II and 46.7% in Group III (*Group II vs. Group III: P Conclusion: The application of CBT has been expanded to the elderly patients (>50 y.o.), although the conditioning regimen and the special medical care for the complications in the early pahse after UCBT has remained to be discussed.

A simple and rapid procedure for sample preparation has been developed for determination of acid coal-tar dyes in confectionery by the matrix solid-phase dispersion (MSPD) method. Confectionery samples spiked with food dyes (12 permitted and 4 non-permitted in Japan) were blended with octadecylsilyl (C18)-derivatized silica packing material pretreated with cetyltrimethylammonium bromide-methanol. A column containing the C18/confectionery matrix was first washed with water, then the dyes were eluted with methanol. The eluate from the column was free from interfering compounds as confirmed by thin-layer and high performance liquid chromatography with UV detection at 254nm. Recoveries of acid coal-tar dyes from confectionery spiked at 100μg/g were 76.7-102.4%.