Jino Son

Endosulfan-Induced Biomarkers in Japanese Rice Fish (Oryzias latipes) Analyzed by SELDI-TOF-MS

The objective of this study was to find and validate estrogen-related biomarkers from plasma proteins in Oryzias latipes after exposure to an estrogen disrupting compound, α-endosulfan. The acute toxicity of α-endosulfan on O. latipes after 96 h of exposure was 13.72, 16.18, and 22.18 μg L-1 for the LC10, LC20, and LC50 values, respectively. To confirm estrogenic disturbance by α-endosulfan, the expression level of vitellogenin in the liver of male fishes was measured at the LC10 value, and it was found to be significantly different from the reference group, confirming the estrogenic effect of endosulfan in this concentration range. Proteinchip® array techniques using a weak cation exchange (CM10) and a strong anion exchange proteinchip (Q10) in conjunction with surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) were used to determine plasma proteins of O. latipes differently expressed in response to endosulfan exposure at LC10 and LC20 concentrations. Analysis of protein profiling of the male fish exposed to α-endosulfan detected 48 significantly different protein peaks and the proteins at m/z 2819, 8462, 8860, and 9462 were significantly different (p<0.05). The protein peaks at m/z 2819, 8860, and 9462 were up-regulated and the peak at m/z 8462 was down-regulated. Therefore, these four differentially expressed proteins could be used as biomarkers to rapidly determine a possible risk of endosulfan on aquatic ecosystems, although these are not necessarily produced as a result of endocrine disruption.

Combined effects of water quality parameters on mixture toxicity of copper and chromium toward Daphnia magna.

In this study, a central composite design (CCD) was employed to evaluate the combined effects of pH, hardness and dissolved organic carbon (DOC) on the toxicity of a mixture of Cu(II) and Cr(VI) toward Daphnia magna. Overall, the results showed that increases in pH, hardness and DOC concentration led to decreased mixture toxicity of Cu(II) and Cr(VI) by reducing the concentrations of toxic species such as Cu(2+) and HCrO(4)(-). In addition, empirical models for the prediction of 24-h and 48-h mortalities of D. magna were developed and validated by using three different sources of dissolved organic matter (DOM). Because the DOMs had different Cu(II)-binding capacities, the empirical models were revised using the ligand concentration of DOMs instead of the DOC concentration; however, the prediction capability of these models did not differ significantly. These results suggest that it is not likely that the chemical property of DOM is important for prediction of the mixture toxicity of Cu(II) and Cr(VI) toward D. magna when the ligand concentration of DOMs greatly exceed the Cu(II) concentration.

Biomarker discovery and proteomic evaluation of cadmium toxicity on a collembolan species, Paronychiurus kimi (Lee)†

The goal of this study was to identify promising new biomarkers of cadmium by identifying differentially expressed proteins in Paronychiurus kimi after exposure to cadmium. Through proteomic analysis of P. kimi using 1‐D PAGE and nano‐LC‐MS/MS, 36 downregulated proteins and 40 upregulated proteins were found. Some of the downregulated and upregulated proteins were verified by LC‐MS/MS analysis after 2‐D PAGE. Downregulated proteins in response to cadmium exposure were involved in glycolysis and energy metabolism, chaperones, transcription, reproduction, and neuron growth. In contrast, proteins involved in glycolysis and energy production, neurogenesis, defense systems response to bacteria, and protein biosynthesis were upregulated in cadmium‐treated collembolans. Cubulin may be a potential biomarker for the detection of cadmium in P. kimi since this biomarker was able to low levels (3.5 mg/kg) of cadmium. The 14‐3‐3 ζ was also found to be a potential biomarker for the detection of medium levels (14 mg/kg) of cadmium. Collembolans may be an alternative tool to humans because many collembolans proteins show a high homology to human proteins.

Response surface model for predicting chronic toxicity of cadmium to Paronychiurus kimi (Collembola), with a special emphasis on the importance of soil characteristics in the reproduction test

A central composite design (CCD) was employed to investigate the effects of organic matter (OM) content and soil pH on the reproduction, and chronic toxicity (28-d EC(50-reproduction)) of cadmium for Paronychiurus kimi after 28days exposure in a standard artificial soil. Two statistical models were developed, one describing reproduction in control artificial soils as a function of OM content and pH, and the other describing cadmium toxicity to the same soil parameters. In the reproduction model, pH was the most important factor, followed by two quadratic factors of OM(2) and pH(2). The parameter pH alone could explain 75.5% of the response variation. The reproduction model will allow us to predict a mean reproduction in the non-treated control soils that contain various combinations of OM content and different pH values. In the chronic toxicity model, only the linear factor of the OM content and pH significantly (p<0.05) affect cadmium toxicity, which explains the 78.9% and 14.9% of total response variance, respectively. Therefore, the final polynomial regression describing the chronic toxicity of cadmium to P. kimi is as follows: predicted 28-d EC(50) of cadmium (mgkg(-1))=-21.231+2.794 x OM+4.874 x pH. The present study show that soil characteristics, which can alter the toxicity of cadmium, can also act as stressors themselves in regards to the reproduction of P. kimi. Based on the physico-chemical characteristics of the test media, the response surface model developed in this study can be used to provide initial toxicity information for cadmium within a region of interest in terms of OM content and pH, and may lead to more scientific based risk assessment for metals.

Joint toxic action of binary metal mixtures of copper, manganese and nickel to Paronychiurus kimi (Collembola)

The joint toxic effects of binary metal mixtures of copper (Cu), manganese (Mn) and nickel (Ni) on reproduction of Paronhchiurus kimi (Lee) was evaluated using a toxic unit (TU) approach by judging additivity across a range of effect levels (10-90%). For all metal mixtures, the joint toxic effects of metal mixtures on reproduction of P. kimi decreased in a TU-dependent manner. The joint toxic effects of metal mixtures also changed from less than additive to more than additive at an effect level lower than or equal to 50%, while a more than additive toxic effects were apparent at higher effect levels. These results indicate that the joint toxicity of metal mixtures is substantially different from that of individual metals based on additivity. Moreover, the close relationship of toxicity to effect level suggests that it is necessary to encompass a whole range of effect levels rather than a specific effect level when judging mixture toxicity. In conclusion, the less than additive toxicity at low effect levels suggests that the additivity assumption is sufficiently conservative to warrant predicting joint toxicity of metal mixtures, which may give an additional margin of safety when setting soil quality standards for ecological risk assessment.

Effect of Soil Organic Matter Content and pH on Toxicity of Cadmium to Paronychiurus kimi (Lee) (Collembola)

Abstract The heterogeneous physico-chemical properties of soil make it difficult to normalize the toxicological effects of metals in the Collembolan species Paronychiurus kimi (Lee). Furthermore, the physico-chemical properties of soil themselves can function as limiting factors that affect population fitness. In this study, the effects of soil organic matter (OM) and pH on the biological performance of P. kimi were assessed, and the influence of these properties on cadmium toxicity to P. kimi was also assessed at the individual and population levels. The OM and pH were found to significantly influence offspring production. All toxicological values of cadmium to P. kimi showed consistent patterns of increase with increasing OM and pH values, implying a decreased toxicity. The 28-d LC50 values varied from 34.9 to 115.9 mg/kg, and the 28-d EC50 and 28-d ri=0 also varied depending on the OM and pH values. These findings indicate that the OM and pH values themselves are important factors in determining not only the biological performance of P. kimi but also the toxicity of cadmium to P. kimi on the individual and population levels. Therefore, when ecological risk assessments are conducted for a certain soil, pre-examinations of the effects of abiotic factors and the selection of an appropriate endpoint must be accomplished before establishing levels of chemicals of concern (COC) for a certain species.

Exposure of mayfly Ephemera orientalis (Ephemeroptera) eggs to heavy metals and discovery of biomarkers

The objective of this study was to assess acute toxicity of heavy metals in eggs of mayfly Ephemera orientalis McLachlan, and to elucidate relationships between heavy metal toxicity and protein expression patterns determined using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Acute toxicity analysis was conducted using five heavy metals (cadmium, chromium, copper, lead, and mercury), and the toxicity endpoint was established from the egg hatching rate during a 14-day exposure period. Median hatching toxicity (HC₅₀) values were determined for each heavy metal, and the most toxic heavy metal was found to be mercury (0.11 mg/L), followed by copper (0.32 mg/L) and lead (4.39 mg/L). E. orientalis eggs were highly tolerant to cadmium and chromium (>120 mg/L). Proteinchip array analysis using a strong anion exchange proteinchip (Q10) in conjunction with SELDI-TOF-MS was used to assess the protein expression patterns after exposure to heavy metals at the EHC10 (prohibiting hatching concentration to 10% eggs), except for cadmium and chromium, which were used at concentrations of 1, 10, and 100mg/L. Three novel biomarker candidate proteins, i.e., 4269, 4283, and 4623 m/z, were identified for the detection of heavy metal toxicity in aquatic ecosystems at the level of HC₁₀ in E. orientalis eggs. SELDI-TOF MS analysis for detecting differential expression of proteins was found to be more effective than Q10 proteinchip separation in the mayfly eggs.

Combined effects of organic matter and pH on acute toxicity of cadmium to Paronychiurus kimi (Collembola): development of response surface model.

A second-order central composite design was employed to investigate the effect of organic matter (OM) content and soil acidity (pH) on the cadmium toxicity to Paronychiurus kimi (Lee), a collembolan species native to Korea. Two independent variables, OM and pH, were adjusted from 0 to 10% (in total dry wt) and 4.5 to 7.0, respectively. Cadmium concentrations tested were 10, 20, 40, 80, and 160 mg/kg dry soil. The toxic effects that cadmium exerted on P. kimi (28-day median lethal concentration (LC50s)) varied significantly with different combinations of OM content and pH. The second-order response surface model was appropriate for describing combined effects of OM and pH on the cadmium toxicity to P. kimi. Linear effects (OM and pH) were predominant contributors accounting for ≈70% of the total variance of cadmium toxicity to P. kimi. No significant quadratic (pH × pH) and interaction (OM × pH) effects were observed; therefore, the final second-order response surface model could be reduced as follows: 28-day LC50 of cadmium (mg/kg) = –94.87 + 26.69 × OM + 16.64 × pH – 1.82 × OM2. Validation of the developed response surface model using 4 soil media with different OM content and pH resulted in a significant correlation coefficient (R2 = 0.97) between predicted and observed 28-day LC50. Based on physico-chemical properties of the test media, developed models can be used to explain the variation of cadmium toxicities at various environmental conditions and may improve current environmental risk assessment procedures of metals.

SU-G-TeP2-15: Feasibility Study of Fiber-Optic Cerenkov Radiation Sensors for in Vivo Measurement: Dosimetric Characterization and Clinical Application in Proton Beams

PURPOSE To evaluate the possibility of a fiber-optic Cerenkov radiation sensor (FCRS) for in vivo dose verification in proton therapy. METHODS The Cerenkov radiation due to the proton beam was measured using a homemade phantom, consisting of a plastic optical fiber (POF, PGSCD1001-13-E, Toray, Tokyo, Japan) connected to each channel of a multianode photomultiplier tube (MAPMT:H7546, Hamamatsu Photonics, Shizuoka, Japan). Data were acquired using a multi-anode photomultiplier tube with the NI-DAQ system (National Instruments Texas, USA). The real-time monitoring graphic user interface was programmed using Labview. The FCRS was analyzed for its dosimetrics characteristic in proton beam. To determine the accuracy of the FCRS in proton dose measurements, we compared the ionization chamber dose measurements using a water phantom. We investigated the feasibility of the FCRS for the measurement of dose distributions near the superficial region for proton plans with a varying separation between the target volume and the surface of 3 patients using a humanoid phantom. RESULTS The dose-response has good linearity. Dose-rate and energy dependence were found to be within 1%. Depth-dose distributions in non-modulated proton beams obtained with the FCRS was in good agreement with the depth-dose measurements from the ionization chamber. To evaluate the dosimetric accuracy of the FCRS, the difference of isocenter dose between the delivery dose calculated by the treatment planning system and that measured by the FCRS was within 3%. With in vivo dosimetry using the humanoid phantom, the calculated surface doses overestimated measurements by 4%-8% using FCRS. CONCLUSION In previous study, our results indicate that the performance of the array-type FCRS was comparable to that of the currently used a multi-layer ion chamber system. In this study, we also believe that the fiber-optic Cerenkov radiation sensor has considerable potential for use with in vivo patient proton dosimetry.

SU-F-T-174: Patient-Specific Point Dose Measurement Using Fiber Optic Radiation Sensor Using Cerenkov Radiation for Proton Therapeutic Beam.

PURPOSE A fiber-optic radiation sensor using Cerenkov radiation (FOCR) has been widely studied for use as a dosimeter for proton therapeutic beam. We developed the FOCR, and it applied to patient-specific point dose measurement in order to evaluate the effectiveness of the FOCR system for proton therapy QA. METHODS Calibration of FOCR was performed with an ionization chamber whose absolute doses were determined according to the IAEA TRS-398 protocol. To determine the calibration curve, the FOCR was irradiated perpendicularly to the proton beam at the 13 dose levels steps. We selected five actual patient treatment plans performed at proton therapy center and compared the resulting FOCR measurements with the ionization chamber measurements. RESULTS The Cerenkov light yield of the FOCR increases linearly with as the dose measured using the ionization chamber increases from 0 cGy to 500 cGy. The results indicate that the fitting curve is linear, suggesting that dose measurement based on the light yield of the FOCR is possible. The results of proton radiation dose QA performed using the FOCR for 10 proton fields and five patients are good agreement with an ionization chamber. CONCLUSION We carried out the patient QA using the FOCR for proton therapeutic beam and evaluated the effectiveness of the FOCR as a proton therapy QA tool. Our results indicate that the FOCR is suitable for use in patient QA of clinical proton beams.