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Variation of Nephrotoxicity Biomarkers by Urinary Storage Condition in Rats

  • Lee, Jung-Min;Han, Young-Hwan;Choi, Su-Jeong;Park, Ju-Seong;Jang, Jeong-Jun;Bae, Re-Ji-Na;Lee, Mi Ju;Kim, Myoung Jun;Lee, Yong-Hoon;Kim, Duyeol;Lee, Hye-Young;Park, Sun-Hee;Park, Cheol-Beom;Kang, Jin Seok;Kang, Jong-Koo
    • Toxicological Research
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    • v.30 no.4
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    • pp.305-309
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    • 2014
  • Recently, there has been an increase in the use of several nephrotoxicity biomarkers in preclinical experiments. In addition, it has been indicated that the result may have been influenced by secondary factors, such as sample storage condition or storage period. In this study, we have assessed the variation in urinary nephrotoxicity biomarkers as a result of urine storage conditions and storage period of the urine. Urine was sampled from specific pathogen-free Sprague-Dawley rats (19 weeks old), which were housed individually in hanged stainless steel wire mesh cages. Urine was stored at $20^{\circ}C$, at $4^{\circ}C$, or at $-70^{\circ}C$ after sampling. The levels of the biomarkers such as beta-2 microglobulin (B2M), cystatin-C (Cys-C), N-acetyl-${\beta}$-D-glucosaminidase (NAG), micro albumin (MA), micro protein (MP) were measured at 6, 24, 48 and 144 hr after sampling. The B2M level was significantly decreased at 6, 24, 48, and 144 hr compared to 0 hr at $-70^{\circ}C$ (p < 0.05, p < 0.01, p < 0.05, and p < 0.05, respectively) and 24 and 144 hr at $20^{\circ}C$ (p < 0.01, p < 0.01, respectively). The Cys-C level was significantly decreased at 144 hr compared to 0 hr at $4^{\circ}C$ (p < 0.01), at $20^{\circ}C$ (p < 0.05) and at $70^{\circ}C$ (p < 0.01). MP and MA levels were not different for 144 hr in all storage conditions. Taken together, B2M and Cys-C levels were modulated by storage temperature and period. For the enhancement of test accuracy, it is suggested that strict protocols be established for samples to minimize the effects of the storage conditions on the detected levels of biomarkers.

Comparison of International Guidelines of Dermal Absorption Tests Used in Pesticides Exposure Assessment for Operators

  • So, Jaehwan;Ahn, Junyoung;Lee, Tae-Hee;Park, Kyung-Hun;Paik, Min-Kyoung;Jeong, Mihye;Cho, Myung-Haing;Jeong, Sang-Hee
    • Toxicological Research
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    • v.30 no.4
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    • pp.251-260
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    • 2014
  • The number of farmers who have suffered from non-fatal acute pesticide poisoning has been reported to vary from 5.7% to 86.7% in South Korea since 1975. Absorption through the skin is the main route of exposure to pesticides for farmers who operate with them. Several in vitro tests using the skins of humans or animal and in vivo tests using laboratory animals are introduced for the assessment of human dermal absorption level of pesticides. The objective of this study is to evaluate and compare international guidelines and strategies of dermal absorption assessments and to propose unique approaches for applications into pesticide registration process in our situation. Until present in our situation, pesticide exposure level to operator is determined just using default value of 10 as for skin absorption ratio because of data shortage. Dermal absorption tests are requested to get exposure level of pesticides and to ultimately know the safety of pesticides for operators through the comparison with the value of AOEL. When the exposure level is higher than AOEL, the pesticide cannot be approved. We reviewed the skin absorption test guidelines recommended by OECD, EFSA and EPA. The EPA recommends assessment of skin absorption of pesticides for humans through the TPA which includes all the results of in vitro human and animal and animal in vivo skin absorption studies. OECD and EFSA, employ a tiered approach, which the requirement of further study depends on the results of the former stage study. OECD guidelines accept the analysis of pesticide level absorbed through skin without radioisotope when the recovery using the non-labeled method is within 80~120%. Various factors are reviewed in this study, including the origin of skin (gender, animal species and sites of skin), thickness, temperature and, etc., which can influence the integrity of results.

Receptor Binding Affinities of Synthetic Cannabinoids Determined by Non-Isotopic Receptor Binding Assay

  • Cha, Hye Jin;Song, Yun Jeong;Lee, Da Eun;Kim, Young-Hoon;Shin, Jisoon;Jang, Choon-Gon;Suh, Soo Kyung;Kim, Sung Jin;Yun, Jaesuk
    • Toxicological Research
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    • v.35 no.1
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    • pp.37-44
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    • 2019
  • A major predictor of the efficacy of natural or synthetic cannabinoids is their binding affinity to the cannabinoid type I receptor ($CB_1$) in the central nervous system, as the main psychological effects of cannabinoids are achieved via binding to this receptor. Conventionally, receptor binding assays have been performed using isotopes, which are inconvenient owing to the effects of radioactivity. In the present study, the binding affinities of five cannabinoids for purified $CB_1$ were measured using a surface plasmon resonance (SPR) technique as a putative non-isotopic receptor binding assay. Results were compared with those of a radio-isotope-labeled receptor binding assay. The representative natural cannabinoid ${\Delta}^9$-tetrahydrocannabinol and four synthetic cannabinoids, JWH-015, JWH-210, RCS-4, and JWH-250, were assessed using both the SPR biosensor assay and the conventional isotopic receptor binding assay. The binding affinities of the test substances to $CB_1$ were determined to be (from highest to lowest) $9.52{\times}10^{-3}M$ (JWH-210), $6.54{\times}10^{-12}M$ (JWH-250), $1.56{\times}10^{-11}M$ (${\Delta}^9$-tetrahydrocannabinol), $2.75{\times}10^{-11}M$ (RCS-4), and $6.80{\times}10^{-11}M$ (JWH-015) using the non-isotopic method. Using the conventional isotopic receptor binding assay, the same order of affinities was observed. In conclusion, our results support the use of kinetic analysis via SPR in place of the isotopic receptor binding assay. To replace the receptor binding affinity assay with SPR techniques in routine assays, further studies for method validation will be needed in the future.

Risk Assessment of 5-Chloro-2-Methylisothiazol-3(2H)-One/2-Methylisothiazol-3(2H)-One (CMIT/MIT) Used as a Preservative in Cosmetics

  • Kim, Min Kook;Kim, Kyu-Bong;Lee, Joo Young;Kwack, Seung Jun;Kwon, Yong Chan;Kang, Ji Soo;Kim, Hyung Sik;Lee, Byung-Mu
    • Toxicological Research
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    • v.35 no.2
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    • pp.103-117
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    • 2019
  • The mixture of 5-chloro-2-methylisothiazol-3(2H)-one (CMIT) and 2-methylisothiazol-3(2H)-one (MIT), CMIT/MIT, is a preservative in cosmetics. CMIT/MIT is a highly effective preservative; however, it is also a commonly known skin sensitizer. Therefore, in the present study, a risk assessment for safety management of CMIT/MIT was conducted on products containing 0.0015% of CMIT/MIT, which is the maximum MIT level allowed in current products. The no observed adverse effect level (NOAEL) for CMIT/MIT was 2.8 mg/kg bw/day obtained from a two-generation reproductive toxicity test, and the skin sensitization toxicity standard value for CMIT/MIT, or the no expected sensitization induction level (NESIL), was $1.25{\mu}g/cm^2/day$ in humans. According to a calculation of body exposure to cosmetics use, the systemic exposure dosage (SED) was calculated as 0.00423 mg/kg bw/day when leave-on and rinse-off products were considered. Additionally, the consumer exposure level (CEL) amounted to $0.77512{\mu}g/cm^2/day$ for all representative cosmetics and $0.00584{\mu}g/cm^2/day$ for rinse-off products only. As a result, the non-cancer margin of safety (MOS) was calculated as 633, and CMIT/MIT was determined to be safe when all representative cosmetics were evaluated. In addition, the skin sensitization acceptable exposure level (AEL)/CEL was calculated as 0.00538 for all representative cosmetics and 2.14225 for rinse-off products; thus, CMIT/MIT was considered a skin sensitizer when all representative cosmetics were evaluated. Current regulations indicate that CMIT/MIT can only be used at concentrations 0.0015% or less and is prohibited from use in other cosmetics products. According to the results of this risk assessment, the CMIT/MIT regulatory values currently used in cosmetics are evaluated as appropriate.

Functional MRI of Visual cortex in the Patients with Occipital Lobe Ischemia (후두엽의 허혈성 뇌졸중 환자에서 시각피질의 기능적 자기공명영상)

  • 이영준;정태섭;윤영수;한승한;조영재;배준호
    • Investigative Magnetic Resonance Imaging
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    • v.3 no.2
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    • pp.173-178
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    • 1999
  • Purpose : To evaluate the usefulness of functional MRI (fMRI) of visual cortex in patients with ischemic infarction in the occipital lobe. Materials and Methods : Four patients with the symptoms and signs of visual cortical ischemia were included. Functional MRI was performed by 2D-FLASH technique with the parameter of 90/56msec TR/TE, $40^{\circ}$ flip angle, $240{\times}240{\;}FOV,{\;}64{\times}128$ matrix number, 8.32 seconds acquisition time, 8mm slice thickness. An axial slice including both visual cortices was selected and alternative activation and resting of the visual cortex was performed using red color photostimulator. all patients undertook visual field test, and vascular abnormality was examined by MRA (n=4) and DSA (n=2). fMRI results were compared with the results of a visual field test, conventional MRI and cerebral angiography. Results : On fMRI, decreased activity of the visual cortex was found in the occipital lobe corresponding to stenosis of the posterior cerebral artery or its branch noted on angiogram. However, 2 of 4 patients showed no abnormal findings on conventional MRI. Visual field defect was noted in 3 patients, one and of whom showed no abnormality on conventional MRI and diffusion-weighted image, but revealed decreased activity in the corresponding visual cortex on fMRI. Conclusion : fMRI may be a sensitive method for detection of the status of decreased blood flow or vascular reserve which other methods can not.

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A Study and Survey on Clinical Nurses concerning the General Items, the Motives of Determining their Profession, the Attitudes toward their Profession and the Desire and Expectation to their Profession and Society (임상간호원에 대한 연구조사)

  • 이귀향;우옥자;서문자
    • Journal of Korean Academy of Nursing
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    • v.3 no.3
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    • pp.78-96
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    • 1973
  • This study of 855 clinical nurses was conducted using a questionnaire that include tour different scales; the motives of determining their profession, the attitudes toward their profession, the general items, and desire and expectation to their profession and society. The data were analyzed by Chi-Square Test and Percentage. The results of this study included Hypothesis are as follows; The respondents were 855 (78.6%) among 1088 clinical nurses who were employed by General of Educational hospitals through the city of Seoul, Pusan, Taegu, Daejun, Kwangju, and Wonju. 1) a. In the Age Distribution, the majority of respondents were under the age of 30yrs(88.2%) and the minority were above 31yrs(11.2%), and the preponderance of the majority to minority(9:1) was noted. In compared with area, a group above 31yrs old in Seoul (6.9%) was lower than other area (16.3%). b. The types of Educational background were 16.3% in Voc.Tr. School, 66.5% in Diploma and 17.1% in Degree.146 clinical nurses were from the Degree course, and 142 (97.3%) CN among those of them were occupied around Seoul and 4(2.7%) around other area. c. In the Marital Status,71,5% were the unmarried and 28.5% were the married. And compared with the area was 20.4% in Seoul and 41.4% in other area. d. Most common Length of Clinical Experience after graduation was under tile 2yrs (55.4%), 3yrs(14.2%, and 4yrs (6.2%). In compared with area, Seoul (15.3%) was lower than other area (38.1%) above 5yrs of clinical experience, and the preponderance of tile other area to Seoul as 2.5: I was noted. 2) a. Hypothesis 1 was significant relation between the types of Educational Background of the CN and their motives for selection of Nursing, P-value was below 0.01. b. There was a significance on hypothesis 2 (P<0.01): that was relation between their motives for selection of clinical nursing field after their graduation and the area which they were employed. c. Hypothesis 4 was accepted as significant relation between the level of satisfaction of their clinical experience after their graduation and the types of educational back ground, P-value was below 0.01. d. There was a significance on hypothesis 5(P<0.01) that was relation between the CN's response about the orientation program and the area which they were employed. e. Hypothesis 6 was retained as significant relation between the area and inservice educational programme of their employed hospital was practising or not. P-value was 0.01. f. Hypothesis 7 was retained as significant relation between the area and the CN's response about the inservice educational programme of their employed. P-value was below 0.01. g. There was a significance on hypothesis 8 (P<0.0l) that was relation between the CN's experience on attending the professional meeting and the area. h. Hypothesis 10 was accepted as significant relation between the response about the present licence system and their educational background. p-value was below 0.01. i. There was a significance on hypothesis 11 (P fO.01) that was relation between the carrying out the regular and delivery vacation and the area. J. Hypothesis 12 was accepted as significant relation between the CN's consideration of the lack of leisure and their marital status. p-value was below 0.01' k. There was a significance on hypothesis 13 (P <0.01) that was relation between the CN's response about their salary and their marital status. l. Hypothesis 14 was significant relation between the most difficulties of CN during their working and the hospital which they were employed. p-value was below 0.01.

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Identification and Functional Analysis of Pig β-1,4-N-Acetylglucosaminyltransferase A (MGAT4A) (돼지 유래의 β-1,4-N-acetylglucosaminyltransferase A (MGAT4A) 유전자의 동정 및 기능 분석)

  • Kim, Ji-Youn;Hwang, Hwan-Jin;Chung, Hak-Jae;Park, Mi-Ryung;Byun, Sung June;Kim, Kyung-Woon
    • Journal of Life Science
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    • v.26 no.3
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    • pp.275-281
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    • 2016
  • Glycan modification is important in pharmaceutical industry. Especially, sialic acid affects the bioactivity and stability of medicine. Milk of pig has been used as bioreactor to produce various pharmaceutical proteins. Therefore, it is necessary to modify the glycan chain in pig mammary grand. β-1,4-N-Acetylglucosaminyltransferase A (pMGAT4A) is one of the essential enzymes for increase of sialic acid content, but pig MGAT4A is unclear. In this study, the pMGAT4A was identified and characterized. The pMGAT4A has 1638 nucleotides encoding 535 amino acids and type II membrane topology, which is one of the common features in many glycosyltransferases. The gene was strongly expressed in liver and mammary gland, whereas was weakly expressed in small intestine, stomach and bladder. For functional test, HA-tagged MGAT4A was over-expressed in porcine kidney (PK-15) cell line. Forced expression of pMGAT4A gene was identified by qPCR, and we identified that pMGAT4A is located in Golgi complex by co- staining with HA antibody and BODIPY TR ceramide. In addition, we identified the increase of mannose-β-1,4-N-acetylglucosamine structure by ELISA and immunofluorescence using Datura stramonium agglutinin (DSA), which recognizes mannose-β-1,4-Nacetylglucosamine. Through the specific activity analysis, we showed that pMGAT4A modified bi-antennary to tri-antennary. This event affects sialic acid content. Therefore, we thought that over-expression of pMGAT4A will be necessary in pig mammary grand for improved medicine.

Diffusion-Weighted MR Imaging of the Brain Tumors: The Clinical Usefulness (두개강내 종양의 확산강조자기공명영상: 임상적 유용성)

  • 이영철;서정진;정광우;강형근;김윤현
    • Investigative Magnetic Resonance Imaging
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    • v.4 no.1
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    • pp.34-41
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    • 2000
  • Purpose: To evaluate the clinical usefulness of diffusion weighted MR imaging(DWI) in the differential diagnosis of brain tumors. Materials and methods: DWI and conventional MR images of nineteen patients with brain tumors(10 metastatic tumors, 4 high grade gliomas, 4 low grade astrocytomas, one oligodendroglioma)were obtained on 1.5T unit. DWI was obtained using single shot spin echo planar imaging with b-value near 1000. We analyzed the signal intensities of lesions including solid portion, necrotic or cystic portion and peritumoral edema of brain tumors (classified five grades comparison with the signal intensities of brain parenchyma and CSF)and calculate the SIR(signal intensity ratio)of lesions to the contralateral normal brain parenchyma. We analyzed statistically the signal intensities and SIR of tumors using independence T test. Results: In solid portions of tumors, all the metastatic tumors and high grade gliomas showed high signal intensities, but low grade astrocytomas and oligodendroglioma showed iso or slight high signal intensities to the normal brain parenchyma. The SIR of solid portion has positive correlation with malignant pot ential(metastatic tumors 1.52, high grade gliomas 1.38, low grade astrocytomas 1.16, oligodendroglioma 1.31)(p < 0.05). In peritumoral edema where seen in 14 tumors, seven of 10 metastatic tumors and two of 4 high grade gliomas showed iso signal intensities, whereas edemas in other 5 brain tumors showed hyperintense to the normal brain parenchyma. The SIRs of peritumoral edemas in metastatic tumors (1.14) was lower than high grade gliomas(1.31),but statistically insignificant. The SIR of cystic or necrotic portion of brain tumors was 0.63. In non enhancing solid portions, three of six cases showed hyperintense to the adjacent peritumoral edema. Conclusion: On DWI, the signal intensities of solid portion has positive correlation with malignant potential, and perilesional edema of brain tumors appear various signal intensities owing to "T2 shine through effect" and the extensiveness of vasogenic edema. Another merit using DWI on the evaluation of brain tumors is to improved better delineation of tumor margins from the adjacent edemas, especially at the non enhancing solid portion of the tumors.

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A High Essential Amino Acid Properties Rice Cultivar 'Haiami' (필수아미노산 고함유 신품종 '하이아미')

  • Hong, Ha-Cheol;Kim, Yeon-Gyu;Yang, Chang-Ihn;Hwang, Hung-Goo;Lee, Jeom-Ho;Lee, Sang-Bok;Choi, Yong-Hwan;Kim, Hong-Yeol;Lee, Kyu-Seong;Yang, Sae-Jun;Kim, Myeong-Ki;Jeong, O-Young;Cho, Young-Chan;Jeon, Yong-Hee;Choi, Im-Soo;Jeong, Eung-Gi;Oh, Sea-Kwan;O, Myeong-Gyu;Yea, Jong-Du;Shin, Young-Seoup;Kim, Jeong-Ju
    • Korean Journal of Breeding Science
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    • v.43 no.6
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    • pp.543-548
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    • 2011
  • Haiami is a new Japonica rice variety developed from a cross between 'Jinmibyeo' TR treated with ethyl methane sulfonate (EMS) EMS and 5-methytryptophan, and 'Gyehwabyeo' in order to develop a new premium quality rice variety by a rice breeding team of National Institute of Crop Science, Rural Development Administration in 2008. This variety has about 138 days of growth duration from transplanting to harvesting in central plain area of Korea. The heading date of this vareity was on $15^{th}$, August. The 'Haiami' has good semi-elect plant type and resistant to lodging with strong culm. The number of panicles/hill of 'Haiami' is more than that of 'Hwaseongbyeo'. This variety shows slow leaf senescence and considerable tolerance to viviparous germination. It is susceptible to leaf blast, bacterial blight, and insect pests, but resistance to rice stripe virus. The milled rice of this variety exhibited translucent, clear non-glutinous endosperm and short grain shape. The essential amino acid properties of 'Haiami' have more than 31% that of 'Hwaseongbyeo' in polished rice. This variety has premium palatability of cooked rice. The yield performance of this rice cultivar was about 5.38 MT/ha in milled rice in local adaptability test for three years from 2006 to 2008. 'Haiami' is adaptable to central and southern plain areas of Korea.