Clinical spectrum of glutamic acid decarboxylase antibodies in a Taiwanese population.

BACKGROUND AND PURPOSE: High levels of autoantibodies against glutamic acid decarboxylase (GAD-abs) are associated with stiff-person syndrome (SPS). However, the full clinical spectrum associated with GAD-abs in Asians is unclear. The clinical and immunological features of patients positive for GAD-abs were reviewed in a large Taiwanese series. METHODS: Retrospective case series and immunological investigations were conducted between July 2007 and July 2017 at a tertiary referral centre in Taiwan. Amongst 361 patients with GAD-ab reactivity, 185 with detailed clinical records were included. RESULTS: Twenty-seven patients (14.59%), with a mean age at assessment of 54.8 ± 13.9 years, presented with neurological symptoms. The major neurological presentations (mean GAD-ab concentrations) were SPS (n = 9, 33.3%; 135.45 ± 27.84 U/ml), cerebellar ataxia (n = 3, 11.1%; 95.61 ± 49.63 U/ml), encephalopathy (n = 2, 7.4%; 51.8 ± 49.64 U/ml) and epilepsy (n = 1, 3.7%; 83.3 U/ml). Notably, eight patients fulfilling the clinical diagnosis of multiple system atrophy had relatively lower GAD-ab concentrations (2.57 ± 0.82 U/ml), which has not been reported previously. There was no correlation between disease severity and GAD-ab concentration. Patients presenting with comorbid endocrinopathies (n = 15, 55.5%) had higher GAD-ab concentrations than those without endocrinopathies (n = 12, 44.4%; 104.45 ± 22.51 U/ml vs. 34.08 ± 21.83 U/ml, P = 0.04). Of 158 patients (85.4%) without a neurological presentation, 133 had type 1 diabetes mellitus and 20 had diabetes of other aetiologies (type 2 or gestational diabetes mellitus, or diabetes secondary to pancreatitis); the remaining four patients had pure thyroid disorders. CONCLUSIONS: A clinical and immunological evaluation of East Asian patients positive for GAD-abs is presented and a different clinical spectrum of anti-GAD syndrome is identified compared to Caucasians.

Acidification and sulfide formation control during reductive dechlorination of 1,2-dichloroethane in groundwater: Effectiveness and mechanistic study.

To enhance the reductive dechlorination of 1,2-dichloroethane (DCA) in groundwater, substrate injection may be required. However, substrate biodegradation causes groundwater acidification and sulfide production, which inhibits the bacteria responsible for DCA dechlorination and results in an odor problem. In the microcosm study, the effectiveness of the addition of ferrous sulfate (FS), desulfurization slag (DS), and nanoscale zero-valent iron (nZVI) on acidification and sulfide control was studied during reductive dechlorination of DCA, and the emulsified substrate (ES) was used as the substrate. Up to 94% of the sulfide was removed with FS and DS addition (0.25 wt%) (initial DCA concentration = 13.5 mg/L). FS and DS amendments resulted in the formation of a metal sulfide, which reduced the hydrogen sulfide concentration as well as the subsequent odor problem. Approximately 96% of the DCA was degraded under reductive dechlorination with nZVI or DS addition using ES as the substrate. In microcosms with nZVI or DS addition, the sulfide concentration was reduced to less than 15 µg/L. Acidification can be controlled via hydroxide ions production after nZVI oxidation and reaction of free CaO (released from DS) with water, which enhanced DCA dechlorination. The quantitative polymerase chain reaction results confirmed that the microcosms with nZVI added had the highest Dehalococcoides population (up to 2.5 × 10(8) gene copies/g soil) due to effective acidification control. The α-elimination mechanism was the main abiotic process, and reductive dechlorination dominated by Dehalococcides was the biotic mechanism that resulted in DCA removal. More than 22 bacterial species were detected, and dechlorinating bacteria existed in soils under alkaline and acidic conditions.

Bioremediation of 1,2-dichloroethane contaminated groundwater: Microcosm and microbial diversity studies.

In this study, the effectiveness of bioremediating 1,2-dichloroethane (DCA)-contaminated groundwater under different oxidation-reduction processes was evaluated. Microcosms were constructed using indigenous bacteria and activated sludge as the inocula and cane molasses and a slow polycolloid-releasing substrate (SPRS) as the primary substrates. Complete DCA removal was obtained within 30 days under aerobic and reductive dechlorinating conditions. In anaerobic microcosms with sludge and substrate addition, chloroethane, vinyl chloride, and ethene were produced. The microbial communities and DCA-degrading bacteria in microcosms were characterized by 16S rRNA-based denatured-gradient-gel electrophoresis profiling and nucleotide sequence analyses. Real-time polymerase chain reaction was applied to evaluate the variations in Dehalococcoides spp. and Desulfitobacterium spp. Increase in Desulfitobacterium spp. indicates that the growth of Desulfitobacterium might be induced by DCA. Results indicate that DCA could be used as the primary substrate under aerobic conditions. The increased ethene concentrations imply that dihaloelimination was the dominate mechanism for DCA biodegradation.

Hybrid algorithms for fuzzy reverse supply chain network design.

In consideration of capacity constraints, fuzzy defect ratio, and fuzzy transport loss ratio, this paper attempted to establish an optimized decision model for production planning and distribution of a multiphase, multiproduct reverse supply chain, which addresses defects returned to original manufacturers, and in addition, develops hybrid algorithms such as Particle Swarm Optimization-Genetic Algorithm (PSO-GA), Genetic Algorithm-Simulated Annealing (GA-SA), and Particle Swarm Optimization-Simulated Annealing (PSO-SA) for solving the optimized model. During a case study of a multi-phase, multi-product reverse supply chain network, this paper explained the suitability of the optimized decision model and the applicability of the algorithms. Finally, the hybrid algorithms showed excellent solving capability when compared with original GA and PSO methods.

Control of trichloroethylene plume migration using a biobarrier system: a field-scale study.

The objective of this field-scale study was to evaluate the effectiveness of controlling trichloroethylene (TCE) plume migration using the polycolloid substrate (PS) biobarrier. The developed PS (containing soybean oil, lactate and surfactants) could release substrate to enhance the TCE dechlorination. In this study, a biobarrier comprising PS injection wells was installed. Injection wells were installed at 5-m intervals, and approximately 15 L of PS was injected into each well. Results show that TCE concentrations in the injection wells dropped from an average of 87 µg/L to below 1 µg/L after 35 days of PS injection. The total organic carbon concentrations in the injection wells increased from an average of 2.1-543 mg/L after 30 days of PS injection. The dissolved oxygen (DO) concentrations and oxidation-reduction potential (ORP) values dropped from an average of 1.6 mg/L to below 0.1 mg/L and from 124 mv to -14 mv after 20 days of injection, respectively. The DO and ORP remained in anaerobic conditions during the remaining 100 days of the operational period. TCE degradation by-products were observed in groundwater samples during the operational period. This reveals that the addition of PS could effectively enhance the reductive dechlorinating of TCE.

The role of sympathetic and parasympathetic nerve systems on the smooth muscle of rat seminal vesicles - experimental results and speculation for physiological implication on ejaculation.

Ejaculation is a process involving sympathetic and parasympathetic effects during different stages - emission and ejection. Some conditions of ejaculation dysfunction are associated with autonomic nerves. However, the exact effects of autonomic nerves on ejaculation are not well defined. Autonomic agonists induce different recorded trace patterns of seminal vesicular contraction. The different traces contain different components of phasic and tonic contraction, which may have physiological implications. In this study, we examined isolated rat seminal vesicle (SV) contraction by phenylephrine (PE), acetylcholine, and their respective antagonists and then speculated upon physiological roles of sympathetic and parasympathetic nerves on SV during ejaculation. We found that PE and Ach both achieved good contraction of rat SV. Compared to α1b for sympathetic and M1, M2 for parasympathetic receptors, α1a and M3 are the relatively dominant subtypes on rat SV. Adrenergic and cholinergic agonists cause different trace patterns of SV contraction. We speculated that the sympathetic effect is dominant during emission to squeeze seminal fluid out and that the parasympathetic effect is dominant during ejection to provide an anti-reflux effect on the ejaculatory duct.

The urodynamic characteristics and prognostic factors of patients with interstitial cystitis/bladder pain syndrome.

AIMS: The role of urodynamic study (UDS) in diagnosis of interstitial cystitis/bladder pain syndrome (IC/BPS) remains inconclusive, nor has the predictor for a treatment success been elucidated. We evaluated the diagnostic and prognostic values of UDS in patients with IC/BPS. MATERIALS AND METHODS: IC/BPS patients with complete data of a symptom assessment, video UDS, potassium chloride (KCl) test, cystoscopic hydrodistention and treatment records were reviewed retrospectively. O'Leary-Sant symptom index and problem index (ICSI, ICPI) were available in part of the patients. The associations between UDS parameters with symptom scores, KCl test result, cystoscopy findings and treatment outcome were analysed. The UDS parameters, including first sensation of filling (FSF), first desire to void (FD), strong desire to void (SD), cystometric bladder capacity (CBC), maximum flow rate (Q(max)), detrusor pressure at Q(max) (Pdet), voided volume (VV) and postvoid residual (PVR). Univariate and multivariate logistic regression analyses were used to identify the variable predicting treatment success. RESULTS: A total of 214 IC/BPS patients were enrolled in this study. The mean volumes at FSF, FD, SD, CBC and VV were inversely correlated with ICSI and ICPI (all p < 0.05). The patients with a positive KCl test had significantly lower mean volumes at SD, CBC, VV and lower Q(max) (all p < 0.05). Significantly lower mean volumes for FSF, FD, SD, CBC and VV were found in patients with higher grade of glomerulation under cystoscopy (all p < 0.01). The anaesthetic maximal bladder capacity (MBC) was positively correlated with FSF, FD, SD, CBC, Q(max) and VV (all p < 0.01). Female gender and the volume at SD were found to be the independent predictors for treatment outcome. CONCLUSIONS: Our study demonstrated that there might be a role of UDS to help in diagnosis and prognostication for IC/BPS. Future prospective investigations are warranted.

Development of a slow polycolloid-releasing substrate (SPRS) biobarrier to remediate TCE-contaminated aquifers.

In this study, an in situ slow polycolloid-releasing substrate (SPRS) biobarrier system was developed to continuously provide biodegradable substrates for the enhancement of trichloroethylene (TCE) reductive dechlorination. The produced SPRS contained vegetable oil (used as a slow-released substrate), cane molasses [used as an early-stage (fast-degradable) substrate], and surfactants [Simple Green (SG) and soya lecithin (SL)]. An emulsification study was performed to evaluate the globule droplet size and stability of SPRS. The distribution and migration of the SPRS were evaluated in a column experiment, and an anaerobic microcosm study was performed to assess the capability of SPRS to serve as a slow and long-term carbon-releasing substrate for TCE dechlorination. The results show that a stable oil-in-water (W/O, 50/50) emulsion (SPRS) with uniformly small droplets (D10, 0.93 µm) has been produced, continuously supplying primary substrates. The emulsion containing the surfactant mixture (with 72 mg/L SL and 71 mg/L SG) had a small absolute value of the zeta potential, which reduced the inter-particle repulsion, leading the emulsion droplets to adhere to one another after collision. The addition of SPRS creates anaerobic conditions and leads to a more complete and thorough removal of TCE through biodegradation and sorption mechanisms.

Novel nanostructured biodegradable polymer matrices fabricated by phase separation techniques for tissue regeneration.

Biomimetic nanostructures have a wide range of applications. In particular, biodegradable polymer nanostructures that mimic the subtleties of extracellular matrix may provide favorable cell interactions. In this study, a co-solvent system was developed to configure a thermodynamically metastable biodegradable polymer solution, from which novel nanostructured matrices subsequently formed via wet phase separation (quaternary) or a combination with thermally induced phase separation. Three-dimensional (3D) nanostructured porous matrices were further fabricated by combination with particle-leaching (100-300µm glucose). The new co-solvent system may generate matrices with reproducible nanostructures from a variety of biodegradable polymers such as poly(d,l-lactide) (PLA), poly(ε-caprolactone) (PCL) and PCL-based polyurethane. In vitro cell culture experiments were performed with mouse pre-osteoblasts (MC3T3-E1) and human bone marrow-derived mesenchymal stem cells (hBM-MSC) to evaluate the osteoinductive potential of PLA nanostructures. The results showed that nanofibrous (<100nm) membranes promoted the bone-related marker gene expression and matrix mineralization of MC3T3-E1 at 14days. Nanofibrous 3D matrices seeded with hBM-MSC without osteogenic induction supplements demonstrated a 2.5-fold increase in bone matrix deposition vs. the conventional microporous matrices after 14 and 21days. Antimicrobial nanofibers were further obtained by plasma-assisted coating of chitosan on PLA nanofibers. This study reveals a platform for fabricating novel biodegradable nanofibrous architecture, with potential in tissue regeneration.

Potential factors that can be used to differentiate between interstitial cystitis/painful bladder syndrome and bladder oversensitivity in women.

AIM: There is considerable overlap between symptoms of interstitial cystitis/painful bladder syndrome (IC/PBS) and bladder oversensitivity, thereby making it difficult to differentiate between the two based on symptoms alone. We investigated factors that could potentially be used to differentiate between IC/PBS and bladder oversensitivity in women. MATERIALS AND METHODS: Video-urodynamic study (VUDS) results in women with lower urinary tract symptoms (LUTS) were retrospectively analysed. Patients classified as having increased bladder sensation (IBS) were selected for analysis. A potassium chloride (KCl) test was performed and pain or urgency elicited was considered positive response. Cystoscopic hydrodistention demonstrating glomerulation was considered diagnosis of IC/PBS; otherwise bladder oversensitivity was diagnosed. LUTS, urodynamic variables and results of the KCl test were used to predict IC/PBS in these women. RESULTS: A total of 405 women with IBS and 272 symptomatic controls with normal VUDS findings were included. Among 227 IBS patients undergoing KCl test, a positive result was found in 190 and a negative test result was found in 37. Characteristic glomerulation was noted in 170 patients, including 165 with a positive and five with a negative KCl test. We found that storage symptoms and pain had a positive predictive value of 45.3% for IC/PBS. When a CBC ≤ 350 ml was added the positive predictive value was 65% for IC/PBS. A combination of storage symptoms, a CBC ≤ 350 ml, a positive KCl test result with an increase of ≥ 2 on the visual analogue pain scale (VAS) provided 100% predictive of IC/PBS. CONCLUSION: A diagnosis of IC/PBS can be made without cystoscopic hydrodistention in women with increased bladder sensation, having storage symptoms, a CBC ≤ 350 ml, a positive KCl test result and a VAS score ≥ 2.

Use of specific gene analysis to assess the effectiveness of surfactant-enhanced trichloroethylene cometabolism.

The objective of this study was to evaluate the effectiveness of in situ bioremediation of trichloroethylene (TCE)-contaminated groundwater using specific gene analyses under the following conditions: (1) pretreatment with biodegradable surfactants [Simple Green™ (SG) and soya lecithin (SL)] to enhance TCE desorption and dissolution, and (2) supplementation with SG, SL, and cane molasses as primary substrates to enhance the aerobic cometabolism of TCE. Polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE), and nucleotide sequence analysis were applied to monitor the variations in specific activity-dependent enzymes and dominant microorganisms. Results show that TCE-degrading enzymes, including toluene monooxygenase, toluene dioxygenase, and phenol monooxygenase, were identified from sediment samples collected from a TCE-spill site. Results from the microcosm study show that addition of SG, SL, or cane molasses can enhance the aerobic cometabolism of TCE. The TCE degradation rates were highest in microcosms with added SL, the second highest in microcosms containing SG, and lowest in microcosms containing cane molasses. This indicates that SG and SL can serve as TCE dissolution agents and act as primary substrates for indigenous microorganisms. Four dominant microorganisms (Rhodobacter sp., Methyloversatilis sp., Beta proteobacterium sp., and Hydrogenophaga pseudoflava) observed in microcosms might be able to produce TCE-degrading enzymes for TCE cometabolic processes.

High case volume of radiation oncologists is associated with better survival of nasopharyngeal carcinoma patients treated with radiotherapy: a multifactorial cohort analysis.

OBJECTIVES: The relationship between physician case volume and patient outcome in patients with head and neck cancers such as nasopharyngeal carcinoma treated by radiotherapy is unknown. This study was designed to investigate the association between the case volume of radiation oncologists and the survival of patients with nasopharyngeal carcinoma. DESIGN: Retrospective cohort study. SETTING: Based on nationwide claims data (National Health Research Insurance Database) in the years 2002-2008. PARTICIPANTS: Newly diagnosed patients with nasopharyngeal carcinoma receiving curative radiotherapy in the year 2003. MAIN OUTCOME MEASURES: Overall survival until 2008. We used the running log-rank test to decide the optimal threshold for categorising the case volume of radiation oncologists. The characteristics of patients, their treatments and contact with health service providers were considered as co-explanatory variables. The log-rank test and Cox regression were performed. Sensitivity analyses were carried out regarding major study assumptions. RESULTS: Five hundred and sixty-two patients with nasopharyngeal carcinoma newly diagnosed in 2003 were identified as the study cohort. The 5-year overall survival was better among patients treated by high-volume (≥6 patients in year 2002) radiation oncologists than by low-volume (<6 patients in year 2002) radiation oncologists (77%versus 64%, P = 0.0007). The adjusted hazard ratio of death was 0.65 (95% confidence interval, 0.48-0.91) upon multivariate analysis. Patients aged at least 65 years also had a lower survival rate than those younger than 65 years old (adjusted hazard ratio of death: 2.81, 95% confidence interval: 1.94-4.08).The physician case volume and patient outcome effect remained the same after sensitivity analyses. CONCLUSIONS: Patients with nasopharyngeal carcinoma treated by high-volume radiation oncologists have better survival compared with those treated by low-volume radiation oncologists. Further studies are needed to verify our findings with similar cancer cohorts treated by modern radiotherapy techniques or other types of radiotherapy.

In situ oxidation of petroleum-hydrocarbon contaminated groundwater using passive ISCO system.

Groundwater contamination by gasoline spill is a worldwide environmental problem. Gasoline contains methyl tertiary-butyl ether (MTBE) (a fuel oxygenates) and benzene, which are the chemicals of concerns among the gasoline components. In this study, an in situ chemical oxidation (ISCO) barrier system was developed to evaluate the feasibility of applying this passive system on the control of MTBE and benzene plume in aquifer. The developed ISCO barrier contained oxidant-releasing materials, which could release oxidants (e.g., persulfate) when contact with water for the contaminants' oxidation in groundwater. In this study, laboratory-scale fill-and-draw experiments were conducted to determine the component ratios of the oxidant-releasing materials and evaluate the persulfate release rates. Results indicate that the average persulfate-releasing rate of 7.26 mg S(2)O(8)(2-)/d/g was obtained when the mass ratio of sodium persulfate/cement/sand/water was 1/1.4/0.24/0.7. The column study was conducted to evaluate the efficiency of in situ application of the developed ISCO barrier system on MTBE and benzene oxidation. Results from the column study indicate that approximately 86-92% of MTBE and 95-99% of benzene could be removed during the early persulfate-releasing stage (before 48 pore volumes of groundwater pumping). The removal efficiencies for MTBE and benzene dropped to approximately 40-56% and 85-93%, respectively, during the latter part of the releasing period due to the decreased persulfate-releasing rate. Results reveal that acetone, byproduct of MTBE, was observed and then further oxidized completely. Results suggest that the addition of ferrous ion would activate the persulfate oxidation. However, excess ferrous ion would compete with organic contaminants for persulfate, and thus, cause the decrease in contaminant oxidation rates. The proposed treatment scheme would be expected to provide a more cost-effective alternative to remediate MTBE, benzene, and other petroleum-hydrocarbon contaminated aquifers. Results from this study will be useful in designing a scale-up system for field application.

Identification and characterization of a novel Rab GTPase-activating protein in spermatids.

We have previously identified novel testis-specific genes by microarray analysis of human testicular tissues. One of the novel genes is Male Germ Cells Rab GTPase- Activating Proteins (MgcRabGAP), which is characterized by the conserved RabGAP catalytic domain, TBC (Tre2/Bub2/Cdc16). RabGAPs are involved in various physiological processes (e.g. vesicular trafficking, cytoskeletal remodelling, cell migration, etc.) by inactivating Rab proteins. In this study, we found that MgcRabGAP transcripts are mainly expressed in the mouse and human testes. The MgcRabGAP protein is expressed in the elongating and elongated spermatids. Immunofluorescence assay of mouse germ cells showed that the protein expression is enriched at the edge of the acrosomal region, neck and annulus during spermiogenesis. This MgcRabGAP is co-localized with its candidate substrate Rab3A at the acrosome/acroplaxome and neck regions of spermatids. Meanwhile, MgcRabGAP is co-localized and interacts with ß-actin. In humans, the expression of MgcRabGAP is enriched at the stage of elongating spermatids. The amount of MGCRABGAP transcript is reduced in the testicular tissues of men with various types of spermatogenic defects. Considering that MGCRABGAP is exclusively expressed in post-meiotic male germ cells, the decreased transcript amount may be a phenomenon secondary to loss of germ cells in the testicular samples. Our finding strongly suggests that MgcRabGAP is involved in acrosome/acroplaxome formation and cytoskeletal reorganization via Rab activity during mammalian spermiogenesis.

Application of persulfate-releasing barrier to remediate MTBE and benzene contaminated groundwater.

The objective of this study was to assess the potential of using an in situ oxidation barrier system to remediate gasoline-contaminated groundwater. The passive remedial system included a persulfate-releasing barrier containing persulfate-releasing materials to release persulfate for contaminant oxidation. Bench experiments were performed to determine the components and persulfate-releasing rate of the persulfate-releasing materials. Column experiments were conducted to evaluate the effectiveness of the designed persulfate-releasing materials on the control of petroleum-hydrocarbon plume. In this study, methyl tert-butyl ether (MTBE) and benzene were used as the target compounds. The optimal persulfate releasing rate was obtained when the mass ratio of persulfate/cement/sand/water was 1/1/0.16/0.5, and the rate varied from 31 to 8 mg persulfate per day per g of material. Significant amounts of MTBE and benzene were removed through the oxidation process due to the release of persulfate, and the produced tert-butyl formate (TBF) and tert-butyl alcohol (TBA), byproducts of MTBE, were further oxidized in the system. Results suggest that the oxidation rate would be affected by the oxidant reduction potential and concentrations of ferrous iron and persulfate.

Evaluation of anti-Wnt/β-catenin signaling agents by pGL4-TOP transfected stable cells with a luciferase reporter system

Refractory and relapsed leukemia is a major problem during cancer therapy, which is due to the aberrant activation of Wnt/β-catenin signaling pathway. Activation of this pathway is promoted by wingless (Wnt) proteins and induces co-activator β-catenin binding to lymphoid enhancer factor (LEF)/T-cell factor protein (TCF). To provide a convenient system for the screening of anti-Wnt/β-catenin agents, we designed a bi-functional pGL4-TOP reporter plasmid that contained 3X β-catenin/LEF/TCF binding sites and a selectable marker. After transfection and hygromycin B selection, HEK 293-TOP and Jurkat-TOP stable clones were established. The luciferase activity in the stable clone was enhanced by the recombinant Wnt-3A (rWnt-3A; 100-400 ng/mL) and GSK3β inhibitor (2’Z,3’E)-6-bromoindirubin-3’-oxime (BIO; 5 µM) but was inhibited by aspirin (5 mM). Using this reporter model, we found that norcantharidin (NCTD; 100 µM) reduced 80 percent of rWnt-3A-induced luciferase activity. Furthermore, 50 µM NCTD inhibited 38 percent of BIO-induced luciferase activity in Jurkat-TOP stable cells. Employing ³H-thymidine uptake assay and Western blot analysis, we confirmed that NCTD (50 µM) significantly inhibited proliferation of Jurkat cells by 64 percent, which are the dominant β-catenin signaling cells and decreased β-catenin protein in a concentration-dependent manner. Thus, we established a stable HEK 293-TOP clone and successfully used it to identify the Wnt/β-catenin signaling inhibitor NCTD.

Evaluation of anti-Wnt/ß-catenin signaling agents by pGL4-TOP transfected stable cells with a luciferase reporter system.

Refractory and relapsed leukemia is a major problem during cancer therapy, which is due to the aberrant activation of Wnt/ß-catenin signaling pathway. Activation of this pathway is promoted by wingless (Wnt) proteins and induces co-activator ß-catenin binding to lymphoid enhancer factor (LEF)/T-cell factor protein (TCF). To provide a convenient system for the screening of anti-Wnt/ß-catenin agents, we designed a bi-functional pGL4-TOP reporter plasmid that contained 3X ß-catenin/LEF/TCF binding sites and a selectable marker. After transfection and hygromycin B selection, HEK 293-TOP and Jurkat-TOP stable clones were established. The luciferase activity in the stable clone was enhanced by the recombinant Wnt-3A (rWnt-3A; 100-400 ng/mL) and GSK3ß inhibitor (2'Z,3'E)-6-bromoindirubin-3'-oxime (BIO; 5 µM) but was inhibited by aspirin (5 mM). Using this reporter model, we found that norcantharidin (NCTD; 100 µM) reduced 80% of rWnt-3A-induced luciferase activity. Furthermore, 50 µM NCTD inhibited 38% of BIO-induced luciferase activity in Jurkat-TOP stable cells. Employing ³H-thymidine uptake assay and Western blot analysis, we confirmed that NCTD (50 µM) significantly inhibited proliferation of Jurkat cells by 64%, which are the dominant ß-catenin signaling cells and decreased ß-catenin protein in a concentration-dependent manner. Thus, we established a stable HEK 293-TOP clone and successfully used it to identify the Wnt/ß-catenin signaling inhibitor NCTD.

Biodistribution of phenylboric acid derivative entrapped lipiodol and 4-borono-2-18F-fluoro-L-phenylalanine-fructose in GP7TB liver tumor bearing rats for BNCT.

A new phenylboric acid derivative entrapped lipiodol (PBAD-lipiodol) was developed as a boron carrier for the boron neutron capture therapy (BNCT) of hepatoma in Taiwan. The biodistribution of both PBAD-lipiodol and BPA-fructose was assayed in GP7TB hepatoma-bearing rat model. The highest uptake of PBAD-lipiodol was found at 2h post injection. The application of BNCT for the hepatoma treatment in tumor-bearing rats is suggested to be 2-4h post PBAD-lipiodol injection.

Seselin from Plumbago zeylanica inhibits phytohemagglutinin (PHA)-stimulated cell proliferation in human peripheral blood mononuclear cells.

Effects of seselin (C(14)H(12)O(3); MW 228) identified from Plumbago zeylanica on phytohemagglutinin (PHA)-stimulated cell proliferation were studied in human peripheral blood mononuclear cells (PBMC). The data demonstrated that seselin inhibited PBMC proliferation-activated with PHA with an IC(50) of 53.87+/-0.74 microM. Cell viability test indicated that inhibitory effects of seselin on PBMC proliferation were not through direct cytotoxicity. The action mechanisms of seselin may involve the regulation of cell cycle progression, interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) production in PBMC. Since cell cycle analysis indicated that seselin arrested the cell cycle progression of activated PBMC from the G(1) transition to the S phase. Seselin suppressed IL-2 and IFN-gamma production in a concentration-dependent manner. Furthermore, seselin significantly decreased the IL-2 and IFN-gamma gene expression in PHA-activated PBMC. Therefore, results elucidated for the first time that seselin is likely an immunomodulatory agent for PBMC.