Impregnation of Activated Carbon with Organic Phase-Change Material.

In this study, we developed a thermal storage medium comprising porous activated carbon filled with organic phase-change materials (PCMs) that utilizes the latent heat of phase-change to absorb heat during heating and release heat during cooling. For the activated carbon, we used both charcoal-based powdered activated carbon (250-350 mesh) and granular activated carbon. The organic phase-change materials used in the experiments were dodecane, tridecane, tetradecane, and pentadecane. Material properties such as thermal conductivity, latent heat, and melting temperature range were evaluated experimentally and theoretically, with the results observed to be consistent. The cyclic thermal performance of the proposed medium was also evaluated. Notably, filling the activated carbon with a mixture of organic PCMs resulted in the highest temperature-moderating effect. The procedure and results presented in this study are expected to aid in further improvement in the performance of thermal storage media containing PCM where stable temperatures are required, including building heating and cooling.

Ternary metal oxide nanocomposite for room temperature H2S and SO2 gas removal in wet conditions.

A ternary Mn-Zn-Fe oxide nanocomposite was fabricated by a one-step coprecipitation method for the remotion of H2S and SO2 gases at room temperature. The nanocomposite has ZnO, MnO2, and ferrites with a surface area of 21.03 m2 g-1. The adsorbent was effective in mineralizing acidic sulfurous gases better in wet conditions. The material exhibited a maximum H2S and SO2 removal capacity of 1.31 and 0.49 mmol g-1, respectively, in the optimized experimental conditions. The spectroscopic analyses confirmed the formation of sulfide, sulfur, and sulfite as the mineralized products of H2S. Additionally, the nanocomposite could convert SO2 to sulfate as the sole oxidation by-product. The oxidation of these toxic gases was driven by the dissolution and dissociation of gas molecules in surface adsorbed water, followed by the redox behaviour of transition metal ions in the presence of molecular oxygen and water. Thus, the study presented a potential nanocomposite adsorbent for deep desulfurization applications.

Behavior of nitrogen and sulfur compounds in the rice husk pellet bioscrubber and its circulation water.

In this study, pellet-type biofilter media was developed with rice husk and applied in a wet scrubber system for odor removal. The lab-scale bioscrubber system was operated for 200 days to evaluate odorous gas removal (i.e., NH3, H2S, methyl mercaptan, and dimethyl sulfide), and the removal mechanism of odor gases was studied by analyzing the behavior of nitrogen and sulfur compounds in circulation water of bioscrubber system. The rice husk pellets supplied the organic carbon source and phosphoric acid necessary for microbial growth, allowing the system to continue successfully for 200 days without any maintenance technology. By analyzing the behavior of the nitrogen and sulfur compounds in the circulation water, we confirmed that the odor gas removal resulted from various mechanisms, including adsorption and biodegradation. Ammonia gas was absorbed by the rice husk pellets and accumulated in the circulation water as nitrite under conditions of sufficient alkalinity and above pH 7. Conversely, when the alkalinity and pH decreased, nitrite was rapidly converted to nitrate. However, H2S gas was oxidized to sulfate and continuously accumulated in the circulation water regardless of the pH and alkalinity. In addition, it was confirmed that the decrease in nitrate in the bioscrubber system was due to heterotrophic denitrification by the organic carbon source supply and autotrophic denitrification by sulfur gas. During the operation of the rice husk pellet bioscrubber for 8 months, under low solubility condition, more than 99% of NH3 and H2S were removed and about 85% of methyl mercaptan (MM) and dimethyl sulfide (DMS) were removed.

Metal-organic framework-derived NaMxOy adsorbents for low-temperature SO2 removal.

This work reported the fabrication of NaMxOy-type adsorbents from air calcination of (Na, M)-trimesate metal-organic frameworks. NaMnxOy (NMO) crystallized as disc-shaped microsheets, whereas NaCoxOy (NCO) crystallized as smooth microsheets with surface deposition of polyhedral nanoparticles. The oxides have a surface area of 1.90-2.56 m2 g-1. The synthesized adsorbents were studied for low-temperature SO2 removal in breakthrough studies. The maximum adsorption capacity of 46.8 mg g-1 was recorded for NMO at 70 °C. The adsorption capacity increased with the increasing temperature due to the chemisorptive nature of the adsorption process. The capacity increased with the increasing bed loading and decreasing flow rate due to the improved SO2 retention time. The elemental mapping confirmed the uniform distribution of sulfur species over the oxide surface. X-ray diffraction showed the absence of metal sulfate nanoparticles in the SO2-exposed samples. The X-ray photoelectron analysis confirmed the formation of surface sulfate and bisulfate. The formation of oxidized sulfur species was mediated by hydroxyl groups over NMO and lattice oxygen over NCO. Thus, the work demonstrated here is the first such report on the use of NaMxOy-type materials for SO2 mineralization.

Toxicity of ZnO/TiO2 -conjugated carbon-based nanohybrids on the coastal marine alga Thalassiosira pseudonana.

Increasing consumption of metal-oxide nanoparticles (NPs) and carbon-based nanomaterials has caused significant concern about their potential hazards in aquatic environments. The release of NPs into aquatic environments could result in adsorption of NPs on microorganisms. While metal-oxide NP-conjugated carbon-based nanohybrids (NHs) may exhibit enhanced toxicity toward microorganisms due to their large surface area and the generation of reactive oxygen species (ROS), there is a lack of information regarding the ecotoxicological effects of NHs on marine diatom algae, which are an indicator of marine pollution. Moreover, there is scant information on toxicity mechanisms of NHs on aquatic organisms. In this study, four NHs (ie, ZnO-conjugated graphene oxide [GO], ZnO-conjugated carbon nanotubes [CNTs], TiO2 -conjugated GO, and TiO2 -conjugated CNT) that were synthesized by a hydrothermal method were investigated for their toxicity effects on a Thalassiosira pseudonana marine diatom. The in vitro cellular viability and ROS formation employed at the concentration ranges of 50 and 100 mg/L of NHs over 72 hours revealed that ZnO-GO had the most negative effect on T. pseudonana. The primary mechanism identified was the generation of ROS and GO-induced dispersion that caused electrostatic repulsion, preventing aggregation, and an increase in surface areas of NHs. In contrast to GO-induced dispersion, large aggregates were observed in ZnO/TiO2 -conjugated CNT-based NHs. The scanning electron microscopy images suggest that NHs covered algae cells and interacted with them (shading effects); this reduced light availability for photosynthesis. Detailed in vitro toxicity effects and mechanisms that cause high adverse acute toxicity on T. pseudonana were unveiled; this implied concerns about potential hazards of these mechanisms in aquatic ecosystems.

Antibacterial effects of graphene- and carbon-nanotube-based nanohybrids on Escherichia coli: Implications for treating multidrug-resistant bacteria.

Some nanomaterials including Fe0, Ag0, and ZnO are well known for their antibacterial effects. However, very few studies have examined antibacterial effects of nanohybrids. Given that metal oxides, mainly ZnO and TiO2, are known to increase mobility, surface area, and photocatalysis when combined with carbon-based nanomaterials, ZnO- and TiO2-conjugated carbon nanotube and graphene oxide nanohybrids were investigated for their antibacterial effects on Escherichia coli (DH5α, a multidrug-resistant coliform bacterium). Graphene-oxide (GO)-based nanohybrids (ZnO-GO and TiO2-GO) induced increased dispersion compared to carbon-nanotube (CNT)-based nanohybrids (ZnO-CNT and TiO2-CNT). Among the four types of nanohybrids, ZnO-conjugated nanohybrids exhibited a higher antibacterial property, resulting in the antibacterial effect (measured with growth inhibition of cells) in the order ZnO-GO > ZnO-CNT > TiO2-GO > TiO2-CNT. Among four possible antibacterial mechanisms (generation of reactive oxygen species (ROS), physicochemical characteristics, the steric effect, and release of metal ions), a primary mechanism-ROS generation-was identified; whereas, physicochemical characteristics and the steric effect were part of contributing mechanisms. The increasing dispersion of TiO2/ZnO on GO may have contributed to the antibacterial effects due to increasing surface areas. Similarly, significant damages to E. coli cell membranes were found by the GO sheet with its sharp edges. Our results suggest that applying GO-based ZnO or TiO2 could be an effective antibacterial method, especially for the treatment of multidrug-resistant bacteria in the water.

Treatment of antibiotic-resistant bacteria by encapsulation of ZnO nanoparticles in an alginate biopolymer: Insights into treatment mechanisms.

Treating multidrug-resistant bacteria has been a challenging task, although the bacteria have been reported as a trace contaminant present in tap water. Given emerging issues on antibiotic-resistant bacteria, the present study investigated a novel treatment method in which ZnO nanoparticles (NPs) are encapsulated in an alginate biopolymer solution to explore primary antibacterial mechanisms. The antibacterial effects of this technology on two model antibiotic-resistant bacteria (Escherichia coli DH5-α and Pseudomonas aeruginosa) were found to be highly effective, with the removal rates of 98% and 88%, respectively, at the initial bacteria concentration of 108 CFU mL-1 over 6 h. The inactivation of antibiotic-resistant bacteria by ZnO NP-alginate beads was improved by increasing the nanocomposite amount (4, 10, and 20 mg) and contact time. The primary mechanism involved the generation of reactive oxygen species (ROS). The ZnO NP-alginate beads were demonstrated to be highly promising for different applications in water treatment, especially for point-of-use in the perspectives of reusability, antibacterial property of ZnO, immobilizing NPs, and utilizing high surface area of NPs, with a slight release of zinc ions.

Effects of coating materials on antibacterial properties of industrial and sunscreen-derived titanium-dioxide nanoparticles on Escherichia coli.

Organic or inorganic stabilizers are often used for coating nanoparticles (NPs) in consumer products. However, upon release of stabilized NPs into the environment, uncertainty exists as to the antimicrobial properties of NPs due to stabilizers and the resultant bioaccumulation in organisms. This study investigates antibacterial effects and subsequent mechanisms of TiO2 NPs on Escherichia coli (E. coli) in the presence and absence of stabilizers (CMC, PVP, and SiO2) commonly used in consumer products. Compared with uncoated TiO2 NPs, the presence of any stabilizers tested in this study increased toxicity of NPs and enhanced growth inhibition in E. coli. While the particle sizes of TiO2 were smaller as the result of coating with PVP or CMC and appeared to contribute to E. coli cell damage, the generation of reactive oxygen species (ROS) was independent of stabilizer type. In fact, coating with PVP and CMC exerted ROS scavenging properties. In contrast, increased ROS production was observed at higher concentrations of TiO2 and upon coating with SiO2. This impact of SiO2 can be related to the formation of a TiOSi chemical bond. The results of the present study emphasize the importance of nanoparticle coating to their anti-bacterial activity and toxicity.

Pilot-scale application of shotblast dust for phosphorus removal.

Phosphorus contamination is a global issue, and cost-effective remediation is sought for removing phosphorus from water. We applied a novel use of waste material called shotblast dust in a pilot-scale reactor to remove phosphorus from water. Results indicate that shotblast dust was effective in treating phosphorus-laden water with 132 kg of the material treating 568 liters of 220 µg/L total phosphorus (T-P) water on a daily basis, achieving approximately 60% removal of T-P in 7 days.

Therapeutic DC vaccination with IL-2 as a consolidation therapy for ovarian cancer patients: a phase I/II trial.

While ovarian cancer (OvCa) responds well to surgery and conventional chemotherapy, a high recurrence rate of advanced OvCa is observed. In this phase I/II study, 10 OvCa patients with minimal residual disease were treated with autologous dendritic cells (DCs) and IL-2 to evaluate the safety and feasibility of this therapeutic strategy and to characterize the antigen-specific immune alterations induced through this treatment. Approximately 4 months after initial debulking and chemotherapy, patients received two subcutaneous doses of autologous monocyte-derived DCs pulsed with autologous tumor lysate and keyhole limpet hemocyanin (KLH) at 4-week intervals. After each DC inoculation, low-dose (200 mIU) IL-2 was introduced for 14 consecutive days as an immune adjuvant. The vaccination was well tolerated. In three out of 10 patients, the inclusion status after the initial therapy showed the maintenance of complete remission (CR) after DC vaccination for 83, 80.9 and 38.2 months without disease relapse. One patient with stable disease (SD) experienced the complete disappearance of tumor after DC vaccination, and this status was maintained for 50.8 months until tumor recurrence. In two patients with partial response (PR) was not responding to DC vaccination and their disease recurred. In the three patients with disease free long-term survival, significant immune alterations were observed, including increased natural killer (NK) activity, IFN-γ-secreting T cells, immune-stimulatory cytokine secretion and reduced immune-suppressive factor secretion after DC vaccination. Thus, in patients with NED status and increased overall survival, DC vaccination induced tumor-related immunity, potentially associated with long-term clinical responses against OvCa.

Experimental test of error-disturbance uncertainty relations by weak measurement.

We experimentally test the error-disturbance uncertainty relation (EDR) in generalized, strength-variable measurement of a single photon polarization qubit, making use of weak measurement that keeps the initial signal state practically unchanged. We demonstrate that the Heisenberg EDR is violated, yet the Ozawa and Branciard EDRs are valid throughout the range of our measurement strength.

Experimental violation and reformulation of the Heisenberg's error-disturbance uncertainty relation.

The uncertainty principle formulated by Heisenberg in 1927 describes a trade-off between the error of a measurement of one observable and the disturbance caused on another complementary observable such that their product should be no less than the limit set by Planck's constant. However, Ozawa in 1988 showed a model of position measurement that breaks Heisenberg's relation and in 2003 revealed an alternative relation for error and disturbance to be proven universally valid. Here, we report an experimental test of Ozawa's relation for a single-photon polarization qubit, exploiting a more general class of quantum measurements than the class of projective measurements. The test is carried out by linear optical devices and realizes an indirect measurement model that breaks Heisenberg's relation throughout the range of our experimental parameter and yet validates Ozawa's relation.

Intra-osseous injection of donor mesenchymal stem cell (MSC) into the bone marrow in living donor kidney transplantation; a pilot study.

BACKGROUND: Mesenchymal stem cells (MSCs) are multi-potent non-hematopoietic progenitor cells possessing an immune-regulatory function, with suppression of proliferation of activated lymphocytes. In this study, adult living donor kidney transplantation (LDKT) recipients were given MSCs derived from the donor bone marrow to evaluate the safety and the feasibility of immunological changes related to the intra-osseous injection of MSC into the bone marrow. METHODS: MSCs were derived from negative HLA cross-match donors. Donor bone marrow was harvested 5 weeks prior to KT. At the time of transplantation, 1 x 106 cell/kg of donor MSC was directly injected into the bone marrow of the recipient's right iliac bone. Patients' clinical outcomes, presence of mixed chimerism by short tandem repeat polymerase chain reaction, analysis of plasma FoxP3 mRNA and cytokine level, and mixed lymphocyte reaction (MLR) were performed. RESULTS: Seven patients enrolled in this study and received donor MSC injections simultaneously with LDKT. The median age of recipients was 36 years (32 ~ 48). The number of HLA mismatches was 3 or less in 5 and more than 3 in 2. No local complications or adverse events such as hypersensitivity occurred during or after the injection of donor MSC. There was no graft failure, but the biopsy-proven acute rejections were observed in 3 recipients during the follow-up period controlled well with steroid pulse therapy (SPT). The last serum creatinine was a median of 1.23 mg/dL (0.83 ~ 2.07). Mixed chimerism was not detected in the peripheral blood of the recipients at 1 and 8 week of post-transplantation. Donor-specific lymphocyte or T cell proliferation and Treg priming responses were observed in some patients. Plasma level of IL-10, a known mediator of MSC-induced immune suppression, increased in the patients with Treg induction. CONCLUSION: Donor MSC injection into the iliac bone at the time of KT was feasible and safe. A possible correlation was observed between the induction of inhibitory immune responses and the clinical outcome in the MSC-kidney transplanted patients. Further research will be performed to evaluate the efficacy of MSC injection for the induction of mixed chimerism and subsequent immune tolerance in KT.

Systemic immune modulation induced by alcoholic beverage intake in obese-diabetes (db/db) mice.

Alcohol over-consumption is generally immunosuppressive. In this study, the effects of single or repetitive alcohol administration on the systemic immunity of db/db mice were observed to clarify the possible mechanisms for the increased susceptibility of obese individuals to alcohol-related immunological health problems. Alcohol (as a form of commercially available 20% distilled-alcoholic beverage) was orally administered one-time or seven times over 2 weeks to db/db mice and normal C57BL/6J mice. Immunologic alterations were analyzed by observation of body weight and animal activity, along with proportional changes of splenocytes for natural killer cells, macrophages, and T and B lymphocytes. Modulation of plasma cytokine level and immune-related genes were also ascertained by micro-bead assay and a microarray method, respectively. The immune micro-environment of db/db mice was an inflammatory state and adaptive cellular immunity was significantly suppressed. Low-dose alcohol administration reversed the immune response, decreasing inflammatory responses and the increment of adaptive immunity mainly related to CD4(+) T cells, but not CD8(+) T cells, to normal background levels. Systemic immune modulation due to alcohol administration in the obese-diabetic mouse model may be useful in the understanding of the induction mechanism, which will aid the development of therapeutics for related secondary diseases.

Chemotherapeutic candidate inducing immunological death of human tumor cell lines.

The immunological death induction by EY-6 on the human tumor cell lines was screened. Human colon carcinoma (HCT15, HCT116), gastric carcinoma (MKN74, SNU668), and myeloma (KMS20, KMS26, KMS34) cells were died by EY-6 treatment with dose-dependent manner. CRT expression, a typical marker for the immunological death, was increased on the EY-6-treated colorectal and gastric cancer cells. Interestingly, the effects on the myeloma cell lines were complicated showing cell line dependent differential modulation. Cytokine secretion from the EY-6 treated tumor cells were dose and cell-dependent. IFN-γ and IL-12 secretion was increased in the treated cells (200% to over 1000% of non-treated control), except HCT116, SNU668 and KMS26 cells which their secretion was declined by EY-6. Data suggest the potential of EY-6 as a new type of immuno-chemotherapeutics inducing tumor-specific cell death. Further studies are planned to confirm the efficacy of EY-6 including in vivo study.

Dendritic Cell (DC) Vaccine in Mouse Lung Cancer Minimal Residual Model; Comparison of Monocyte-derived DC vs. Hematopoietic Stem Cell Derived-DC.

The anti-tumor effect of monocyte-derived DC (MoDC) vaccine was studied in lung cancer model with feasible but weak Ag-specific immune response and incomplete blocking of tumor growth. To overcome this limitation, the hematopoietic stem cell-derived DC (SDC) was cultured and the anti-tumor effect of MoDC & SDC was compared in mouse lung cancer minimal residual model (MRD). Therapeutic DCs were cultured from either CD34(+) hematopoietic stem cells with GM-CSF, SCF and IL-4 for 14 days (SDC) or monocytes with GM-CSF and IL-4 for 7 days (MoDC). DCs were injected twice by one week interval into the peritoneum of mice that are inoculated with Lewis Lung Carcinoma cells (LLC) one day before the DC injection. Anti-tumor responses and the immune modulation were observed 3 weeks after the final DC injection. CD11c expression, IL-12 and TGF-ß secretion were higher in SDC but CCR7 expression, IFN-γ and IL-10 secretion were higher in MoDC. The proportion of CD11c(+)CD8a(+) cells was similar in both DC cultures. Although both DC reduced the tumor burden, histological anti-tumor effect and the frequencies of IFN-γ secreting CD8(+) T cells were higher in SDC treated group than in MoDC. Conclusively, although both MoDC and SDC can induce the anti-tumor immunity, SDC may be better module as anti-tumor vaccine than MoDC in mouse lung cancer.

Combination therapy of renal cell carcinoma or breast cancer patients with dendritic cell vaccine and IL-2: results from a phase I/II trial.

BACKGROUND: Ten cancer patients (Six renal cell carcinoma and four breast cancer patients) were treated in a phase I/II study with a vaccine composed of autologous dendritic cells (DCs) and IL-2 to evaluate the DC vaccine-related toxicity and antigen-specific immune alteration. METHODS: Cancer patients were treated twice with autologous CD34+ hematopoietic stem cell-derived, GM-CSF/IFN-γ-differentiated DCs pulsed with autologous tumor lysate and KLH, by 4-week interval. Following each subcutaneous injection of therapeutic DCs, low-dose (200 MIU) IL-2 was introduced for 14 consecutive days as an immune adjuvant. To determine the DC vaccine-induced immunological alterations, the KLH-specific lymphocyte proliferation, number of IFN-γ secreting T cells (ELISPOT assay), NK activity and the cytokine modulation were measured. RESULTS: Cultured-DCs expressing HLA-DR, CD11c, CD83, and B7.1/B7.2 produced IL-12p70. After vaccination, the patients tolerated it. Clinical response was observed in one RCC patient as stable disease. However DC-vaccine related antigen-specific immune responses including peripheral blood lymphocyte proliferation and the number of IFN-r secreting cells were induced in six patients without clear correlation with clinical responses. Also NK activity was induced significantly in six patients after vaccination. DC vaccine-related decrease of TGF-ß level or increase of IL-12p70 level and decline of CD4+CD25+ T cells were observed in three patients. However only in the RCC patient whose disease stabilized, combination of stimulatory as well as inhibitory immune alterations including induction of IFN-γ secreting T cell with reduction of CD4+ CD25+ T cell were correlated with clinical responses. CONCLUSION: Data indicated that DC vaccine combined with IL-2 is well tolerated without major side effects. DC vaccine induced the specific immunity against introduced antigen. Combinatorial alterations of immunological parameters indicating antigen-specific immune induction along with reduction of inhibitory immunity were correlated with clinical responses in DC vaccine treated patients.

Cellular Mechanism of Newly Synthesized Indoledione Derivative-induced Immunological Death of Tumor Cell.

BACKGROUND: EY-6 is one of the newly synthesized indoledione derivatives to induce tumor cell-specific cell death. In this study, we investigated the mechanism of immunological death induced by EY-6 at mouse colon cancer cell as well as at the normal immune cell represented by dendritic cell. METHODS: C57BL/6 mouse syngeneic colon cancer cell MC38 was treated with EY-6, and analyzed by MTT for viability test, flow cytometry for confirming surface expressing molecules and ELISA for detection of cytokine secretion. Normal myeloid-dendritic cell (DC) was ex vivo cultured from bone marrow hematopoietic stem cells of C57BL/6 mice with GM-CSF and IL-4 to analyze the DC uptake of dead tumor cells and to observe the effect of EY-6 on the normal DC. RESULTS: EY-6 killed the MC38 tumor cells in a dose dependent manner (25, 50 and 100 µM) with carleticulin induction. And EY-6 induced the secretion of IFN-γ but not of TNF-α from the MC38 tumor cells. EY-6 did not kill the ex-vivo cultured DCs at the dose killing tumor cells and did slightly but not significantly induced the DC maturation. The OVA-specific cross-presentation ability of DC was not induced by chemical treatment (both MHC II and MHC I-restricted antigen presentation). CONCLUSION: Data indicate that the EY-6 induced tumor cell specific and immunological cell death by modulation of tumor cell phenotype and cytokine secretion favoring induction of specific immunity eliminating tumor cells.

Nonlocal dispersion cancellation using entangled photons.

A pair of optical pulses traveling through two dispersive media will become broadened and, as a result, the degree of coincidence between the optical pulses will be reduced. For a pair of entangled photons, however, nonlocal dispersion cancellation in which the dispersion experienced by one photon cancels the dispersion experienced by the other photon is possible. In this paper, we report an experimental demonstration of nonlocal dispersion cancellation using entangled photons. The degree of two-photon coincidence is shown to increase beyond the limit attainable without entanglement. Our results have important applications in fiber-based quantum communication and quantum metrology.

Phase I/II study of immunotherapy using autologous tumor lysate-pulsed dendritic cells in patients with metastatic renal cell carcinoma.

This phase I/II study was conducted to evaluate the feasibility, safety and efficacy of immunotherapy using tumor lysate (TL)-pulsed dendritic cells (DC) in patients with metastatic renal cell carcinoma (RCC). DC were generated by culturing peripheral blood mononuclear cells in the presence of GM-CSF and IL-4 and were pulsed with autologous TL and keyhole limpet hemocyanin (KLH). Maturation of DC was induced by a combined treatment of CD40 ligand, IFN and monocyte-conditioned medium. The patients were administered two cycles of TL-pulsed DCs vaccination, each of which comprised of four doses injected subcutaneously at biweekly intervals. Nine patients were included. The immunotherapy was well tolerated without severe side effects. One patient achieved an objective partial response (PR). Five patients showed stable disease (SD), and the remaining three had progressive disease (PD). With a median follow-up of 17.5 months, the median time to progression was 5.2 months and the median overall survival was 29 months. In the antigen-specific lymphocyte proliferation assay, eight patients showed a proliferative response, which tended to be stronger in patients with SD or PR than in patients with PD. The ELISPOT assay was performed in two patients and indicated that one patient with PR showed a much stronger response than another with PD. Our results suggest that TL-pulsed DC immunotherapy in combination with nephrectomy affect the natural course of RCC and are associated with clinical benefits for patients with metastatic diseases.