A proinflammatory cytokine sensor cell for assaying inflammatory activities of nanoparticles.
Explore the source record for details and available documents.
Adult↗
Biomedical subjects
Sung Bae Kim
Publications and source records attributed to Sung Bae Kim.
A method for determining the activities of cytokines based on the nuclear transport of nuclear factor-kappaB.
Explore the source record for details and available documents.
Active Transport, Cell Nucleus↗
Treatment-induced menstrual changes in very young (<35 years old) breast cancer patients.
PURPOSE: To evaluate the treatment-induced menstrual changes in very young (< 35 years old) breast cancer patients. METHODS AND MATERIALS: We retrospectively examined the clinical records of 160 patients, ranging in age from 18 to 34 years old (median age, 32 years), treated between June 1992 and December 2002. One hundred twenty patients underwent mastectomy and 40 underwent breast conserving surgery. Postoperatively, 80 patients were treated with alkylating agent-based chemotherapy regimens (CMF) and 80 with anthracycline-based regimens (AD). In addition, 57 patients received adjuvant radiotherapy, and 77 received anti-estrogen therapy. Treatment-induced menstrual changes and present menstrual status were evaluated from hospital records and by one-to-one interviews. The median follow-up period was 54 months (range, 29-156 months). RESULTS: Treatment-induced menstrual change (amenorrhea) was occurred in 59 (36.9%) patients, 25 (31.3%) of those treated with CMF and 34 (42.5%) with AD (p=0.142). Amenorrhea occurred after a median 2 cycles of chemotherapy (range, 1-6 cycles). Menstruation resumed in 49 (83.1%) patients, 20 (80%) of those treated with CMF and 29 (85.3%) with AD (p=0.6). Median time to resumption of menstruation was median 3.5 months (range, 1-18 months) after amenorrhea. Disease recurred in 10 (16.9%) patients who experienced treatment-induced menstrual changes and in 18 (17.8%) of those who did not (p=0.89). CONCLUSION: Although the overall incidence of treatment-induced menstrual change in breast cancer patients under age 35 was similar to that reported elsewhere, the rate of recovery from these change is higher. We observed no difference between CMF and AD treated patients in rates of amenorrhea or recovery from these changes.
Adolescent↗
Enhancement of the enzymatic digestibility of waste newspaper using Tween.
Methods of increasing the enzymatic digestibility of waste newspaper by adding Tween (TW)-20 and 80 surfactants were investigated. Tween-series surfactants were selected because these surfactants increase cellulase activity during enzymatic hydrolysis and do not inhibit cell growth in downstream fermentation processes. When surfactant was used in a pretreatment, a benefic effect was expected in the enzymatic hydrolysis stage owing to surfactant carry-over from the pretreatment stage immediately upstream of the hydrolysis. However, because it was necessary to wash the pretreated substrate with water to remove inhibitors produced during pretreatment, no added benefit was obtained. When surfactant was used in the pretreatment only, it was found that it had a marked effect on digestibility and that this effect was higher at lower enzyme loadings. Also, TW-80 was found to be more effective than TW-20, and the addition of enzyme and TW-80 to substrate at the beginning of enzyme reaction was found to most effectively increase digestibility. When TW-80 was added into either the pretreatment stage or the hydrolysis stage the digestibilities of untreated sample increased by approx 40%, whereas an increase of only 45% was observed when TW-80 was added to both stages. These results show that the addition of surfactant to either the pretreatment or the enzymatic hydrolysis stage is sufficient to increase digestibility.
Cellulase↗
Quantitative determination of protein nuclear transport induced by phosphorylation or by proteolysis.
Nucleocytoplasmic transport of proteins in eukaryotic cells is a fundamental process for gene expression. The transport is regulated by posttranslational modifications of the proteins, such as ligand-binding, phosphorylation, and proteolysis. For monitoring the nuclear transport of proteins induced by a ligand binding, we have recently developed a genetically encoded bioluminescent indicator based on reconstitution of split fragments of Renilla reniformis (RLuc) by protein splicing with DnaE inteins. We herein describe that the technique is used for detecting phosphorylation- or proteolysis-induced nuclear transports of a target protein. Two model proteins, signal transducer and activator of transcription 3 (STAT3) and sterol-regulatory element binding protein-2 (SREBP-2), were exemplified as phosphorylation- and proteolysis-induced nuclear transport, respectively. Each STAT3 or SREBP-2 is connected with C-terminal halves of RLuc and DnaE. If the protein translocates into the nucleus, the C-terminal fragment of RLuc meets the N-terminal fragment of RLuc, and full-length RLuc is reconstituted by protein splicing in the nucleus. The indicator with SREBP-2 enabled us to quantify the intracellular concentrations of cholesterol. The indicator with STAT3 quantified the extent of the nuclear transport induced by representative cytokines. This simple assay based on protein nuclear transports allows the selection of suitable drugs among candidates and has significant potential for risk assessments, such as carcinogenic chemical screening in vitro and in vivo.
Active Transport, Cell Nucleus↗
Genetically encoded stress indicator for noninvasively imaging endogenous corticosterone in living mice.
Physical and emotional stress is one of the major controllers of physiological reactions and homeostasis in living animals. A stress hormone, corticosterone, is secreted from the adrenal cortex into the blood vessels when animals sense the stress. The quantitative evaluation of corticosterone in living animals has been limited because of the unavailability of suitable methods in vivo. For a noninvasive molecular imaging of the stress, we developed a method for detecting physiological increases in the endogenous corticosterone caused by exo- and endogenous stress in living animals. We constructed a pair of genetically encoded indicators composed of cDNAs of glucocorticoid receptor (GR), split Renilla luciferase (RLuc), and a Synechocystis sp. DnaE intein. The GR fused with C-terminal halves of RLuc and DnaE is localized in the cytosol, whereas a fusion protein of N-terminal halves of RLuc and DnaE is localized in the nucleus. If corticosterone induces GR translocation into the nucleus, the C-terminal RLuc meets the N-terminal one in the nucleus, and full-length RLuc is reconstituted by protein splicing with DnaE. Cell-based methods provided a quantitative bioluminescence assay of the extent of GR translocation into the nucleus. We further demonstrated that the indicator enabled noninvasive imaging against two different types of imposed stress: a forced swimming and metabolic perturbation caused by 2-deoxy-D-glucose. This stress indicator should be valuable for screening pharmacological compounds and for tools to study the mechanism of physiological stress.
Animals↗
A genetically encoded indicator for assaying bioactive chemicals that induce nuclear transport of glucocorticoid receptor.
Glucocorticoids, the adrenal steroid hormones secreted during stress, are essential to homeostasis and metabolism in the human body. An impaired glucocorticoid signaling due to dysfunction of the glucocorticoid receptor (GR) by synthetic chemicals can cause diseases and disruptions of the homeostasis and metabolism. Here we demonstrate the development of a method for screening endocrine-disrupting chemicals and potent risk factors of human diseases based on the nuclear trafficking of the GR. We constructed a new assay using a pair of genetic indicators with the full length of the GR, split Renilla luciferase (RLuc), and split DnaE (a protein splicing element). The GR-containing fusion protein with C-terminal halves of DnaE and RLuc is localized in cytosol due to the cytosolic character of the GR, whereas the fusion protein with N-terminal halves of DnaE and RLuc stays in the nucleus due to the cofused nucleus localization signal. On being stimulated with a ligand, the GR is translocated into the cellular nucleus. Thus, a protein splicing occurs in the nucleus by an interaction between the splicing junctions of each DnaE fragment. The enzymatic activities from the reconstituted RLuc allow the ligand-dependent luminescence intensities. The feasibility of the method was evaluated by quantifying the hormonal activities of 20 different kinds of steroids and synthetic chemicals using the NIH 3T3 cells carrying the pair of indicators. The hormonal activities of tested ligands are discussed based on the chemical structure-activity relationship. We found that androgens, testosterone, and 19-nortestosterone weakly induce the nuclear transport of the GR. The current assay allows high-throughput screening of risk chemicals and drug candidates influential to a signal transduction pathway of the GR.
Active Transport, Cell Nucleus↗
FDG-PET in the prediction of pathologic response after neoadjuvant chemoradiotherapy in locally advanced, resectable esophageal cancer.
PURPOSE: To assess the efficacy of 18Fluorodeoxyglucose-positron emission tomography (FDG-PET) for predicting a pathologic response in locally advanced esophageal cancer after neoadjuvant chemoradiotherapy. METHODS AND MATERIALS: All enrolled patients were treated with neoadjuvant chemoradiotherapy followed by esophagectomy and underwent two FDG-PET scans, before and after neoadjuvant chemoradiotherapy. We compared the results of the preoperative FDG-PET scans with the pathologic results. RESULTS: From July 2001 to July 2004, 32 patients (29 men and 3 women) were enrolled in this study. Pathologic complete response (pCR) in the esophagus was achieved in 21 of 32 patients (66%). The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) in the primary tumors of the preoperative FDG-PET were 27%, 95%, 75%, and 71%, respectively. In regional lymph nodes, these values were 16%, 98%, 36%, and 93%, respectively. The mean standardized uptake value (SUV) of primary tumors was initially 5.6 +/- 3.6 and changed to 1.5 +/- 1.3 after neoadjuvant chemoradiotherapy (p < 0.05). If analysis of metabolic response (SUV decrease, DeltaSUV) was limited to initially highly metabolic primary tumors (SUV > or =4.0), pathologic response was correlated with metabolic response (p = 0.006). CONCLUSIONS: This study suggested that the pathologic response of an initially highly metabolic tumor after neoadjuvant chemoradiotherapy could be correlated with the metabolic response, and FDG-PET can provide additional information on tumor response to chemoradiotherapy.
Aged↗
High-throughput sensing and noninvasive imaging of protein nuclear transport by using reconstitution of split Renilla luciferase.
Nucleocytoplasmic trafficking of functional proteins plays a key role in regulating gene expressions in response to extracellular signals. We developed a genetically encoded bioluminescent indicator for monitoring the nuclear trafficking of target proteins in vitro and in vivo. The principle is based on reconstitution of split fragments of Renilla reniformis (Rluc) by protein splicing with a DnaE intein (a catalytic subunit of DNA polymerase III). A target cytosolic protein fused to the N-terminal half of Rluc is expressed in mammalian cells. If the protein translocates into the nucleus, the Rluc moiety meets the C-terminal half of Rluc, and full-length Rluc is reconstituted by protein splicing. We demonstrated quantitative cell-based in vitro sensing of ligand-induced translocation of androgen receptor, which allowed high-throughput screening of exo- and endogenous agonists and antagonists. Furthermore, the indicator enabled noninvasive in vivo imaging of the androgen receptor translocation in the brains of living mice with a charge-coupled device imaging system. These rapid and quantitative analyses in vitro and in vivo provide a wide variety of applications for screening pharmacological or toxicological compounds and testing them in living animals.
Active Transport, Cell Nucleus↗
Hand-foot syndrome in patients treated with capecitabine-containing combination chemotherapy.
Clinical characteristics and risk factors of hand-foot syndrome were investigated in patients who received capecitabine-containing chemotherapy. Toxicity data were analyzed from 179 patients in 4 prospective clinical trials testing docetaxel/capecitabine/cisplatin in stomach cancer, capecitabine/cisplatin in biliary or stomach cancer, and vinorelbine/capecitabine in breast cancer. Hand-foot syndrome was reported in 116/179 (64.8%) of patients, with grade 3 hand-foot syndrome in 8/179 (4.5%). Hand-foot syndrome first developed within the first 3 chemotherapy cycles in 100/116 (86.2%) patients, with the median onset for all 3 treatment regimens occurring during cycle 2. Because severe reactions were rare, hand-foot syndrome was not a major factor influencing treatment schedule. Risk factor analyses showed that combined use of docetaxel and preceding chemotherapy-related stomatitis were significant risk factors for the development of hand-foot syndrome. Our results suggest that a combined treatment agent and a patient's susceptibility to chemotherapy-related toxicity may increase the risk of capecitabine-induced hand-foot syndrome.
Adult↗
Enhancement of enzymatic digestibility of recycled newspaper by addition of surfactant in ammonia-hydrogen peroxide pretreatment.
The effect of surfactant on enzymatic digestibility was investigated during the pretreatment stage. Newspaper was pretreated with an ammonia-hydrogen peroxide mixture on a shaking bath at 40 degrees C and 130 strokes/min for 3 h. Two kinds of nonionic surfactants, NP series and Tween series, were utilized. The effect of hydrophile-lipophile balance (HLB) value of both series surfactants on digestibility was found to be negligible, even though de-inking efficiency was improved as HLB value was increased. The effect of surfactant loading on digestibility was small, below 0.5 wt%, and negligible above 0.5 wt% at 60 international filter paper units (IFPU). The percentage improvement in digestibility increased as enzyme loading decreased. Digestibility of NP-5-added sample relative to control sample, increased significantly at an enzyme loading <60 IFPU, i.e., 19 and 13% at 15 and 30 IFPU, respectively. Such an increase in digestibility was not explained clearly from the experimental results. It was also found that ink removal before enzymatic hydrolysis is very important to enhance digestibility.
Ammonia↗
Murine bone marrow stromal cells: implications for their use in gene modified cell therapy.
The objectives of this study were to demonstrate that bone marrow stromal cells (BMSC) can be an attractive novel target of genetic modification in gene and cell therapy of hematologic disease. Here, we investigated the therapeutic effects of gene modified BMSC using a murine lymphoma model. BMSC of Balb/c AnN mice were encoded with the human IL-2 gene (hIL-2) using an adenoviral vector. About 1 x 10(6) cells of a murine B lymphoma (A20) cell line, were injected into the mice via tail vein. One week after injection of A20 cells, the mice were divided into 4 groups for BMSC therapy: No BMSC (control), unmodified BMSC, BMSC with Ad/deltaE1, and BMSC with Ad/hIL-2. Mice were observed for 8 weeks. The results demonstrated that all mice treated with BMSC (Ad/hIL-2) survived with no evidence of disease during this period of observation. All mice treated with unmodified BMSC or BMSC (Ad/deltaE1) as well as the controls developed disseminated lymphoma, and 80% of mice survived less than 4 weeks. In conclusion, the IL-2 gene-modified stromal cell is a promising therapeutic tool for murine lymphoma.
Adenoviridae↗
Enzymatic digestibility of used newspaper treated with aqueous ammonia-hydrogen peroxide solution.
Wastepaper constitutes approximately half of municipal solid waste, making it a potential source of bioenergy. Newspaper was pretreated with an ammonia-hydrogen peroxide (H2O2) mixture in a shaking bath from room temperature to 80 degrees C, and then its enzymatic digestibility was measured. A significant amount of ink was removed from the newspaper slurry by the reciprocating movement of the shaking bath. In addition, the ammonia-H2O2 significantly swelled the substrate, thereby greatly increasing its susceptibility to enzymatic digestion. After pretreating the newspaper with conditions of 40 degrees C, 3 h, 130 strokes/min, and 4 wt% ammonia-2 wt% H2O2, the enzymatic digestibility was almost 90% of theoretical, or about 25% higher than that of untreated substrate. Digestibility was also investigated as a function of ammonia concentration, H2O2 concentration, shaking speed, pretreatment temperature, and time.
Ammonia↗
A screening method for estrogens using an array-type DNA glass slide.
A new screening assay was described for the determination of endocrine disrupting chemicals (EDCs), such as synthetic estrogens, with an array-type DNA glass slide having characteristics of 1) a high sample throughput, 2) a compact size allowing a small sample volume, and 3) a sensitive determination based on the estrogen-dependent binding of the human estrogen receptor a (hERalpha) with its estrogen responsive element (ERE; Vit. A2 gene promoter). We devised a glass slide on which a thin agarose gel was mounted. Avidin was then covalently immobilized on each well of the glass slide after the gel was activated by a NaIO4 solution. Also, the biotinylated ERE as a DNA probe was immobilized on the gel layer through avidin-biotin binding. After the estrogen-dependent binding of a yellow fluorescent protein-fused hERalpha (YFP-hERalpha) to ERE on the gel layer, the fluorescence intensity of YFP-hERalpha quantitatively extracted into the gel was directly determined with a fluorescence microplate reader. Pre-incubation of YFP-hERalpha with estrogen at 37 degrees C for 30 min enhanced the estrogen-dependent hERalpha-ERE binding. The determined hormonal activities of estrogens on the interaction of YFP-hERalpha with ERE were as follows in their decreasing order: diethylstilbestrol (DES) > 17beta-estradiol (E2) <==> ethynylestradiol (EE2) > 4-hydroxy tamoxifen (OHT) > clomiphene (Clo). The present method provides a sensitive estrogen-dependent dose-response curve down to approximately 10(-13) M in the case of DES. This method will become a competitive alternative to the conventional in vitro assays, such as a DNA-binding assay using radioisotopes.
Avidin↗
Polymerase chain reaction-based diagnosis of bone marrow involvement in 170 cases of non-Hodgkin lymphoma.
BACKGROUND: Up to the current time, diagnosis of bone marrow (BM) involvement in non-Hodgkin lymphoma (NHL) has been based on morphologic findings. Polymerase chain reaction (PCR) for antigen receptor gene rearrangements has the potential to increase the detection sensitivity of minimal degrees of BM involvement. The authors therefore assessed PCR-based clonalities of BM concurrently with morphology from 170 cases with NHL and evaluated the usefulness of comparative analysis of clonalities between bilateral BMs and the lymph node and the clinical significance of PCR based clonalities of BM. METHODS: Bilateral BM clot sections of 170 cases and 47 lymph nodes were tested for immunoglobulin heavy chain gene rearrangement or T-cell receptor gamma gene rearrangement according to the B- or T-lineage of the lymph node. RESULTS: When compared with morphology, the results of PCR showed an unexpectedly low positive concordance rate of 61.0% for B-cell NHL and 57.1% for T-cell NHL. When the clonality of BM was compared with that of lymph nodes in B-cell NHL, bilateral clonalities of BM showed high concordance with the clonality of the lymph nodes. PCR-based clonality did not show significant impact on survival. CONCLUSIONS: Morphology remains the gold standard in the evaluation of BM involvement by NHL. Although the comparative analysis of BM clonality and that of the lymph nodes is considered a valuable tool that increases the reliability of clonality, PCR-based clonality of BM does not significantly add to the sensitivity of diagnosing BM involvement by NHL.
Adolescent↗
Diffusion of sulfuric acid within lignocellulosic biomass particles and its impact on dilute-acid pretreatment.
Intra-particle diffusion of sulfuric acid into sugarcane bagasse, corn stover, rice straw and yellow poplar was investigated to determine the effective diffusivity of sulfuric acid within the porous biomass structure. Diffusion experiments were conducted over 25-75 degrees C for two different biomass sizes using dynamic diffusion test cells. Diffusivities of sulfuric acid in agricultural residues were significantly higher than those of hard wood. Diffusivity data for each biomass were fitted into the Arrhenius equation for extrapolation to higher temperatures. The diffusivity data were subsequently incorporated into a theoretical model to determine acid profile within the biomass matrix. The modeling results indicate that intra-particle diffusion of acid influences the rate of dilute-acid pretreatment if unground biomass feedstock is used under normal pretreatment conditions. A criterion was set up to determine the critical biomass size at which the intra-particle acid diffusion becomes a rate-influencing factor for a given pretreatment condition.
Acids↗