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Oral Toxicity Study and Skin Sensitization Test of a Cricket

  • Ryu, Hyeon Yeol;Lee, Somin;Ahn, Kyu Sup;Kim, Hye Jin;Lee, Sang Sik;Ko, Hyuk Ju;Lee, Jin Kyu;Cho, Myung-Haing;Ahn, Mi Young;Kim, Eun Mi;Lim, Jeong Ho;Song, Kyung Seuk
    • Toxicological Research
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    • v.32 no.2
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    • pp.159-173
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    • 2016
  • Crickets have been attracting considerable interest in the field of nutrition and toxicology due to the global exhaustion of food resulting from a growing population. The cricket is normally eaten in several countries after roasting, similar to the grasshopper; however, safety evaluation data on cricket powder is limited. Here, we performed general toxicity studies of cricket powder including a single, 2-week repeated dose range evaluation test, a 13-week repeated oral dose toxicity test in Sprague-Dawley rats, a single oral dose toxicity test in Beagle dogs, and a skin sensitization test in guinea pigs following the Organization for Economic Cooperation and Development test guidelines 406 and 408 in addition to Good Laboratory Practice. To investigate the NOAEL and target organs of cricket powder, Sprague-Dawley rats were allocated to 4 groups: vehicle control, 1,250 mg/kg, 2,500 mg/kg, 5,000 mg/kg dose test groups and cricket powder was administered over 13 weeks after single dose and dose range finding studies in rats based on the results of the single oral administration toxicity study in rats and Beagle dogs. The results of the study showed that the NOAEL of cricket powder was over 5,000 mg/kg for both sexes of rats without adverse effects in a 13-week repeated oral toxicity study and there was no skin hypersensitivity reaction. Therefore, our results reveal that crickets can be widely used as a new substitute food or nutrient resource.

Comparison of In Vitro Cell Transformation Assay Using Murine Fibroblasts and Human Keratinocytes

  • Ahn, Jun-Ho;Park, Sue-Nie;Yum, Yung-Na;Kim, Ji-Young;Lee, Michael
    • Toxicological Research
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    • v.24 no.1
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    • pp.37-44
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    • 2008
  • The in vitro cell transformation assays (CTA) were performed using BALB/3T3 murine fibroblasts and HaCaT human keratinocytes in order to evaluate concordance between both in vitro CTAs and carcinogenicity with compounds differing in their genotoxic and carcinogenic potential. Six test articles were evaluated, two each from three classes of compounds: genotoxic carcinogens (2-amino-5-nitrophenol and 4-nitroquinoline-N-oxide), genotoxic noncarcinogens (8-hydroxyquinoline and benzyl alcohol), and nongenotoxic carcinogens (methyl carbamate and N-nitrosodiphenylamine). Any foci of size $\geq$2 mm regardless of invasiveness and piling was scored as positive in CTA with BALB/3T3. As expected, four carcinogens regardless of their genotoxicity had positive outcomes in two-stage CTA using BALB/3T3 cells. However, of the two genotoxic noncarcinogens, benzyl alcohol was positive CTA finding. We concluded that, of the 6 chemicals tested, the sensitivity for BALB/3T3 system was reasonably high, being 100%. The respective specificity for BALB/3T3 assay was 50%. We also investigated the correlation between results of BALB/3T3 assay and results from HaCaT assay in order to develop a reliable human cell transformation assay. However, evaluation of staining at later time points beyond the confluency stage did not yield further assessable data because most of HaCaT cells were detached after $2{\sim}3$ days of confluency. Thus, after test article treatment, HaCaT cells were split before massive cell death began. In this modified protocol for this HaCaT system, growing attached colonies were counted instead of transformed foci 3 weeks since last subculture. Compared to BALB/3T3 assay, HaCaT assay showed moderate low sensitivity and high specificity. Despite these differences in specificity and sensitivity, both cell systems did exhibit same good concordance between in vitro CTA and rodent carcinogenicity findings (overall 83% concordant results). At present the major weakness of these in vitro CTA is lack of validation for regulatory acceptance and use. Thus, more controlled studies will be needed in order to be better able to assess and quantitatively estimate in vitro CTA data.

Gene Expression Profiling in Diethylnitrosamine Treated Mouse Liver: From Pathological Data to Microarray Analysis (Diethylnitrosamine 처리 후 병리학적 결과를 기초로 한 마우스 간에서의 유전자 발현 분석)

  • Kim, Ji-Young;Yoon, Seok-Joo;Park, Han-Jin;Kim, Yong-Bum;Cho, Jae-Woo;Koh, Woo-Suk;Lee, Michael
    • Toxicological Research
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    • v.23 no.1
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    • pp.55-63
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    • 2007
  • Diethylnitrosamine (DEN) is a nitrosamine compound that can induce a variety of liver lesions including hepatic carcinoma, forming DNA-carcinogen adducts. In the present study, microarray analyses were performed with Affymetrix Murine Genome 430A Array in order to identify the gene-expression profiles for DEN and to provide valuable information for the evaluation of potential hepatotoxicity. C57BL/6NCrj mice were orally administered once with DEN at doses of 0, 3, 7 and 20 mg/kg. Liver from each animal was removed 2, 4, 8 and 24 hrs after the administration. The histopathological analysis and serum biochemical analysis showed no significant difference in DEN-treated groups compared to control group. Conversely, the principal component analysis (PCA) profiles demonstrated that a specific normal gene expression profile in control groups differed clearly from the expression profiles of DEN-treated groups. Within groups, a little variance was found between individuals. Student's t-test on the results obtained from triplicate hybridizations was performed to identify those genes with statistically significant changes in the expression. Statistical analysis revealed that 11 genes were significantly downregulated and 28 genes were upregulated in all three animals after 2 h treatment at 20 mg/kg. The upregulated group included genes encoding Gdf15, JunD1, and Mdm2, while the genes including Sox6, Shmt2, and SIc6a6 were largely down regulated. Hierarchical clustering of gene expression also allowed the identification of functionally related clusters that encode proteins related to metabolism, and MAPK signaling pathway. Taken together, this study suggests that match with a toxicant signature can assign a putative mechanism of action to the test compound if is established a database containing response patterns to various toxic compounds.

Single and Four-Week Repeated Oral Toxicity Study of Antidiabetic Herb Extract Microcapsule in Sprague-Dawley Rats (항당뇨 한약추출고형물의 Sprague-Dawley 랫드를 이용한 단회 및 4주 반복투여 독성시험)

  • Kim, Young-Chul;Kim, Hye-Jeong;Kong, Min-Kyu;Lim, Ae-Kyoung;Kwon, Mi-Hwa;Kim, Kil-Soo;Lee, Gee-Dong
    • Toxicological Research
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    • v.23 no.1
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    • pp.87-96
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    • 2007
  • Single and repeated-dose toxicity of anti-diabetic herb extract microcapsule (ADHEM) were evaluated according to Toxicity Test Guidelines of Korea Food and Drug Administration using Sprague-Dawley rats. For single-dose toxicity test, kneading ADHEM with sterilized water were administered orally once at dose levels of 0 and 2,000 mg/kg and examined for 14 days. No dead animals, clinical signs and abnormal necropsy findings were observed and also no significant difference in body weights was found. Therefore, the $LD_{50}$ of ADHEM was considered to be higher than 2,000 mg/kg in both male and female rats. For repeated-dose toxicity test, ADHEM were mixed with powder fodder and administerd orally for 28 days at dose levels of 0, 500, 1000 and 2000 mg/kg/day. No dead animals, clinical signs and significant difference in body weights were found. In hematology and serum biochemistry, all values were included within the normal ranges. In relative organ weights, kidney or liver were significantly increased in the 500, 1000 or 2000 mg/kg/day male groups, uterus was significantly increased in the 500 mg/kg/day female group and left adrenal glands were significantly decreased in the 2000 mg/kg/day female group. In histopathological examinations, vacuolation and microgranuloma in the liver, chronic progressive nephropathy and inflammation in the kidney were observed in the 500, 1000 or 2000 mg/kg/day both male and female groups. Therefore, the no observed adverse effect level (NOAEL) of ADHEM was considered to be lower than 500 mg/kg/day in both male and female rats.

Comparison of Toxicity and Deposition of Nano-Sized Carbon Black Aerosol Prepared With or Without Dispersing Sonication

  • Kang, Mingu;Lim, Cheol-Hong;Han, Jeong-Hee
    • Toxicological Research
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    • v.29 no.2
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    • pp.121-127
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    • 2013
  • Nanotoxicological research has shown toxicity of nanomaterials to be inversely related to particle size. However, the contribution of agglomeration to the toxicity of nanomaterials has not been sufficiently studied, although it is known that agglomeration is associated with increased nanomaterial size. In this study, we prepared aerosols of nano-sized carbon black by 2 different ways to verify the effects of agglomeration on the toxicity and deposition of nano-sized carbon black. The 2 methods of preparation included the carbon black dispersion method that facilitated clustering without sonication and the carbon black dispersion method involving sonication to achieve scattering and deagglomeration. Male Sprague-Dawley rats were exposed to carbon black aerosols 6 hr a day for 3 days or for 2 weeks. The median mass aerodynamic diameter of carbon black aerosols averaged $2.08{\mu}m$ (for aerosol prepared without sonication; group N) and $1.79{\mu}m$ (for aerosol prepared without sonication; group S). The average concentration of carbon black during the exposure period for group N and group S was $13.08{\pm}3.18mg/m^3$ and $13.67{\pm}3.54mg/m^3$, respectively, in the 3-day experiment. The average concentration during the 2-week experiment was $9.83{\pm}3.42mg/m^3$ and $9.08{\pm}4.49mg/m^3$ for group N and group S, respectively. The amount of carbon black deposition in the lungs was significantly higher in group S than in group N in both 3-day and 2-week experiments. The number of total cells, macrophages and polymorphonuclear leukocytes in the bronchoalveolar lavage (BAL) fluid, and the number of total white blood cells and neutrophils in the blood in the 2-week experiment were significantly higher in group S than in normal control. However, differences were not found in the inflammatory cytokine levels (IL-$1{\beta}$, TNF-${\alpha}$, IL-6, etc.) and protein indicators of cell damage (albumin and lactate dehydrogenase) in the BAL fluid of both group N and group S as compared to the normal control. In conclusion, carbon black aerosol generated by sonication possesses smaller nanoparticles that are deposited to a greater extent in the lungs than is aerosol formulated without sonication. Additionally, rats were narrowly more affected when exposed to carbon black aerosol generated by sonication as compared to that produced without sonication.

Correlation between Instrument on Pattern Identification for Depression and Psychological Tests by Statistical Analysis (통계적 분석을 통한 우울증 변증도구와 심리검사의 상관성 연구)

  • Kim, Hwan;Lee, Hun-Soo;Lee, Eun Jung;Park, Joon-Ho;Kang, Wee-Chang;Jung, In Chul
    • Journal of Oriental Neuropsychiatry
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    • v.27 no.3
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    • pp.131-146
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    • 2016
  • Objectives: This study was performed to determine the correlation between Instrument on Pattern Identification for Depression and Psychological tests by Pearson Correlation Coefficient and Regression analysis.Methods: Two assessors carried out the evaluation using the instrument on pattern identification for depression. They also performed the following psychological tests: Beck Depression Inventory (BDI), State-Trait Anxiety Inventory (STAI), State-Trait Anger Expression Inventory (STAXI), the Temperament and Character Inventory (TCI), Insomnia Severity Index (ISI_Total), Self-disclosure Inventory, subjective well-being Inventory, Health perception Inventory, and Personality Assessment Inventory (PAI). A total of 167 participants who got HAM-D score over 12 were targeted for the evaluation. Our research team carried out Pearson correlation coefficient analyses and regression analysis between pattern identification for depression and Psychological tests. We listed the results by descending order and interpreted the results.Results: Pearson correlation analysis revealed the following results: 1) Stagnation of Liver Gi was associated with BDI (0.60) and STAI (0.55); 2) Dual Deficiency of the Heart and Spleen was associated with BDI (0.60), ISI_Total (0.52), and STAI (0.42); 3) Relieving stagnation of Phlegm-Gi was associated with BDI (0.65), STAI (0.54), and Subjective well-being inventory (−0.52); 4) Gi-deficiency Mingled with sputum was associated with BDI (0.50), ISI_Total (0.40), and STAI (0.395); 5) Stagnant Gi transforming into fire was associated with BDI (0.56), STAI_TR (0.51), and Health perception Inventory (−0.458); 6) Yin-Deficiency with Effulgent Fire was associated with BDI (0.55), ISI_total (0.54), and STAI (0.41).Conclusions: Through correlation analysis between Instrument on Pattern Identification for Depression and Psychological tests, we could suggest a System for Oriental Medical Diagnosis for Depression.

Evaluation of Embryotoxic Potential of Olaquindox and Vitamin A in Micromass Culture and in Rats

  • Kang, Hwan-Goo;Ku, Hyun-Ok;Jeong, Sang-Hee;Cho, Joon-Hyoung;Son, Seong-Wan
    • Toxicological Research
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    • v.26 no.3
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    • pp.209-216
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    • 2010
  • Limb bud (LB) and central nerve system (CNS) cells were prepared from 12.5 day old pregnant female Crj:CD (SD) rats and treated with olaquindox and vitamin A. Cytotoxicity and inhibition on differentiation were measured in each cell. Three doses of olaquindox (4, 21 and 100 mgkg), and 0.2 and 75 mg/kg of vitamin A were administered to pregnant rat for 11 days from $6^{th}$ to $16^{th}$ of pregnancy. $IC_{50}$ values of olaquindox for proliferation and differentiation in CNS cell were 22.74 and $28.32\;{\mu}g/ml$ and 79.34 and $23.29\;{\mu}g/ml$ in LB cell and those values of vitamin A were 8.13 and $5.94\;{\mu}g/ml$ in CNS cell and 0.81 and $0.05\;{\mu}g/ml$ in LB cell, respectively. Mean body weights of pregnant rats were decreased at high dose of olaquindox (110 mg/kg) but relative ovary weight, number of corpus lutea, and number of implantation were not changed. Resorption and dead fetus were increased at high dose of olaquindox, and relative ovary weight, the number of corpus lutea and implantation, and sex ratio of male to female were not significantly changed in all dose of olaquindox. Mean fetal and placenta weights were significantly (p < 0.01) decreased in rats of high group. Seven fetuses out of 103 showed external anomaly like bent tail, and 10 out of 114 fetuses showed visceral anomalies at high group. The ossification of sternebrae and metacarpals were significantly (p < 0.01) increased by low and middle dose of olaquindox but it was significantly (p < 0.01) prohibited by high dose of olaquindox. In rats treated with vitamin A, the resorption and dead fetus were increased by high dose. Mean fetal weights were significantly (p < 0.01) increased by low dose but significantly (p < 0.01) decreased by high dose. Thirty four fetuses out of 52 showed external anomaly; bent tail (1), cranioarchschisis (14), exencephaly (14), dome shaped head (22), anophthalmia (15), brcahynathia (10) and others (19). Forty five fetuses out of 52 showed soft tissue anomaly; cleft palate (42/52) and anophthalmia (22/52) by high dose of vitamin A. Sixty one fetuses out of 61 (85.2%) showed skull anomaly; defect of frontal, partial and occipital bone (21/61), defect of palatine bone (52/61) and others (50/61). In summary, we support that vitamin A is strong teratogen based on our micromass and in vivo data, and olaquindox has a weak teratogenic potential in LB cell but not in CNS cell. We provide the in vivo evidence that a high dose of olaquindox could have weak embryotoxic potential in rats.

Epidemiological Patterns of Cancer Incidence in Southern China: Based on 6 Population-based Cancer Registries

  • Liu, Jie;Yang, Xu-Li;Li, Ai;Chen, Wan-Qing;Ji, Lu;Zhao, Jun;Yan, Wei;Chen, Yi-Ying;Zhu, Li-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1471-1475
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    • 2014
  • Background: The epidemiological patterns of cancer incidence have been investigated widely in western countries. Nevertheless, information is quite limited in Jiangxi province, southern China. Materials and Methods: All data were reported by 6 population-based cancer registries in Jiangxi Province. The results were presented as incidence rates of cases by site (ICD-10), sex, crude rate (CR), age-standardized rates (ASRs) and truncated incidence rate (TR) per 100,000 person-years, using the direct method of standardization to the world population. Results: 8,765 new cancer cases were registered in our study during the period 2009-2011. Diagnosis of cancer was based on histopathology in 61.0%, clinical or radiology findings in 4.87% and death certificate only (DCO) in 3.0% of the cases. The median age at diagnosis was 62.0 years (mean, 61; standard deviation, 15). The ASRs were 170.8 per 100,000 for men and 111.2 for women. The ASRs for all invasive cancers from the urban areas (145.7 per 100,000) was higher than that of rural areas (137.1). Incidence rates for lung cancer were higher in rural (35.8) than in urban areas (27.0). Similarly, relatively high rates were observed for stomach cancer in rural (20.1) relative to urban areas (15.5). Conclusions: Our results reveal that the most common cancers were breast and lung in women and lung and liver in men. Interestingly, this study suggested a higher incidence rates for lung and stomach cancer in rural males than in urban population, which may suggest other potential causes, such as over-consumption of smoked meats and high prevalence of Helicobacter pylori infection, respectively. Public education and the promotion of healthy lifestyles should be actively carried out.

Seasonal Variation of Transparency in the Southeastern Yellow Sea (황해 남동해역 투명도의 계절 변화)

  • CHOI Yong-Kyu;KWON Jung-No
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.3
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    • pp.323-329
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    • 1998
  • The data of Secchi disc observation collected during $1966\~1990$ were analyzed to investigate the seasonal variation of transparency in the southeastern Yellow Sea. The bimonthly distribution of mean transparency showed that the isolines of transparency were roughly parallel to the isobaths. The transparency was low (3 m in february and 8 m in August) if the shallow water less than 20 m depth in comparition to the higher values (10 m in february and 17 m in August) in the deeper water. The lowest transparency was found in winter. The transparency increased in spring and the highest transparency occurred in summer. The water becomes turbid in autumn. Suspended solid concentrations in winter are ranged from 28 to 130 $mg/{\ell}$, and from 8 to 60 $mg/{\ell}$ in summer. The seasonal variation of transparency seems to be mainly affected by resuspension of solid from the bottom. The amounts of suspended solid are large in winter due to the vertical convection by cooling effect and tubulence by the strong wind, and small in summer due to the strong stratification and weak wind.

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Neural Activation in the Somatosensory Cortex by Electrotactile Stimulation of the Fingers: A Human fMRI Study

  • Seok, Ji-Woo;Jang, Un-Jung;Sohn, Jin-Hun
    • Journal of the Ergonomics Society of Korea
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    • v.33 no.5
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    • pp.395-405
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    • 2014
  • Objective: The aim of this study is to investigate 1) somatotopic arrangement of the second and third fingers in SI area 2) difference of neural activation in the SI area produced by stimulation with different frequencies 3) correlation between the intensity of tactile perception by different stimulus intensity and the level of brain activation measurable by means of fMRI. Background: Somatosensory cortex can obtain the information of environmental stimuli about "where" (e.g., on the left palm), "what" (e.g., a book or a dog), and "how" (e.g., scrub gently or scrub roughly) to organism. However, compared to visual sense, the neural mechanism underlying the processing of specific electrotactile stimulus is still unknown. Method: 10 right-handed subjects participated in this study. Non-painful electrotactile stimuli were delivered to two different finger tips of right hand. Functional brain images were collected from 3.0T MRI using the single-shot EPI method. The scanning parameters were as follows: TR and TE were 3000, 35ms, respectively, flip angle 60, FOV $24{\times}24cm$, matrix size $64{\times}64$, slice thickness 4mm (no gap). SPM5 was used to analyze the fMRI data. Results: Significant activations produced by the stimulation were found in the SI, SII, the subcentral gyrus, the precentral gyrus, and the insula. In all participants, statistically significant activation was observed in the contralateral SI area and the bilateral SII areas by the stimulation on the fingers but ipsilaterally dominant. The SI area representing the second finger generally located in the more lateral and inferior side than that of the third finger across all the subjects. But no difference in brain area was found for the stimulation of the fingers by different frequencies. And two typical patterns were observed on the relationship between the perceived psychological intensity and the amount of voxels in the primary sensory cortex during the stimulation. Conclusion: It was possible to discriminate the representation sites in the SI by electrotactile stimulation of digit2 and digit3. But we could not find the differences of the brain areas according to different stimulation frequencies from 3 to 300Hz. Application: The results of the study can provide a deeper understanding of somatosensory cortex and offer the information for tactile display for blinds.