• Journal of Dairy Science and Biotechnology
• /
• v.34 no.2
• /
• pp.99-116
• /
• 2016

We herein performed animal safety assessment in accordance with Good Laboratory Practice (GLP) regulations with the aim of developing sialic acid from glycomacropeptide (hereafter referred to as "GMP") as an index ingredient and functional component in functional foods. GMP is a type of whey protein derived from milk and a safe food, with multiple functions, such as antiviral activity. A test substance was produced containing 7% (w/w) sialic acid and mostly-hydrolyzed whey protein (hereafter referred to as "7%-GNANA") by enzymatic treatment of substrate GMP. The maximum intake test dose level was selected based on 5,000 mg/kg/day dose set for male NOEL (no-observed-effect-level) and female NOAEL (no-observed-adverse-effect-level) determined by a dose-range finding (DRF) test (GLP Center of Catholic University of Daegu, Report No. 15-NREO-001) that was previously conducted with the same test substance. To evaluate the toxicity of a repeated oral dose of the test substance in connection with the previous DRF study, 1,250, 2,500, and 5,000 mg/kg of the substance were administered by a probe into the stomachs of 6-week-old SPF Sprague-Dawley male and female rats for 90 d. Each test group consisted of 10 male and 10 female rats. To determine the toxicity index, all parameters, such as observation of common signs; measurements of body weight and food consumption; ophthalmic examination; urinalysis, electrolyte, hematological, and serum biochemical examination; measurement of organ weights during autopsy; and visual and histopathological examinations were conducted according to GLP standards. After evaluating the results based on the test toxicity assessment criteria, it was determined that NOAEL of the test substance, 7%-GNANA, was 5,000 mg/kg/day, for both male and female rats. No animal death was noted in any of the test groups, including the control group, during the study period, and there was no significant difference associated with test substance, as compared with the control group, with respect to general symptoms, body weight changes, food consumption, ophthalmic examination, urinalysis, hematological and serum biochemical examination, and electrolyte and blood coagulation tests during the administration period (P<0.05). As assessed by the effects of the test substance on organ weights, food consumption, autopsy, and histopathological safety, change in kidney weight as an indicator of male NOAEL revealed up to 20% kidney weight increase in the high-dose group (5,000 mg/kg/day) compared with the change in the control group. However, it was concluded that this effect of the test substance was minor. In the case of female rats, reduction of food consumption, increase of kidney weight, and decrease of thymus weight were observed in the high-dose group. The kidney weight increased by 10.2% (left) and 8.9% (right) in the high-dose group, with a slight dose-dependency compared with that of the control group. It was observed that the thymus weight decreased by 25.3% in the high-dose group, but it was a minor test substance-associated effect. During the autopsy, botryoid tumor was detected on the ribs of one subject in the high-dose group, but we concluded that the tumor has been caused by a naturally occurring (non-test) substance. Histopathological examination revealed lesions on the kidney, liver, spleen, and other organs in the low-dose test group. Since these lesions were considered a separate phenomenon, or naturally occurring and associated with aging, it was checked whether any target organ showed clear symptoms caused by the test substance. In conclusion, different concentrations of the test substance were fed to rats and, consequently, it was verified that only a minor effect was associated with the test substance in the high-dose (5,000 mg/kg/day) group of both male and female rats, without any other significant effects associated with the test substance. Therefore, it was concluded that NOAEL of 7%-GNANA (product name: Helicobactrol) with male and female rats as test animals was 5,000 mg/kg/day, and it thus was determined that the substance is safe for the ultimate use as an ingredient of health functional foods.

• Tuberculosis and Respiratory Diseases
• /
• v.50 no.2
• /
• pp.182-195
• /
• 2001

Background : Idiopathic pulmonary fibrosis(IPF) is a devastating illness for which there is little effective treatment. The key cytokines currently implicated in the fibrotic process are the transforming growth factor-${\beta}_1$(TGF-${\beta}_1$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), endothelin-1(ET-1) and interferon-$\gamma$(IFN-$\gamma$). The rat model for paraquat-induced pulmonary fibrosis was chosen to investigate the role of ET-1 in this disease. Both ET-1 and TGF-${\beta}_1$ expression in lung lesions were examined using immunohistochemical staining. After $Bosentan^{(R)}$ administration, an orally active ET-$l_A$ and ET-$1_B$ receptor antagonist, the degree of pulmonary fibrosis and ET-1 and TGF-${\beta}_1$ expression were analyzed. Method : Sprague-Dawley rats were divided into three groups, the control group, the fibrosis group, and the fibrosis-$Bosentan^{(R)}$-treated group. The animals were sacrificed periodically at 1, 3, 5, 7, 10, 14 days after administering saline or paraquat. The effects between groups were compared with the results of light microscopy and immunohistochemical staining for ET-1 and TGF-${\beta}_1$. The degree of fibrosis was evaluated by H&E and Masson's trichrome staining, which were graded by a computerized image analyzer. The degree of immunohistochemical staining was categorized by a semi-quantitative analysis method. Results : The lung collagen content had increased in the paraquat instillated animals by day 3, and continued to increase up to day 14. A daily treatment by gavage with $Bosentan^{(R)}$ (100mg/kg) did not prevent the increase in collagen deposition on the lung that was induced by paraquat instillation. There were increased immunohistochemical stains of ET-1 on the exudate, macrophages, vascular endothelial cells and pneumocytes in the paraquat instillated group. Furthermore, TGF-${\beta}_1$ expression was higher on the exudate, macrophages, some inflammatory cells, pneumocytes( type I, and II), vascular endothelium and the respiratory epithelial cells around the fibrotic area. After Bosentan treatment, there were no definite changes in ET-1 and TGF-${\beta}_1$ expression. Conclusion : Fibrosis of the Paraquat instillated group was more advanced when compared with the control group. In addition, there was increased ET-1 and TGF-${\beta}_1$ expression around the fibrotic area. ET-1 is associated with lung fibrosis but there was little effect of the ET-1 receptor blocker($Bosentan^{(R)}$) on antifibrosis.Background : Idiopathic pulmonary fibrosis(IPF) is a devastating illness for which there is little effective treatment. The key cytokines currently implicated in the fibrotic process are the transforming growth factor-${\beta}_1$(TGF-${\beta}_1$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), endothelin-1(ET-1) and interferon-$\gamma$(IFN-$\gamma$). The rat model for paraquat-induced pulmonary fibrosis was chosen to investigate the role of ET-1 in this disease. Both ET-1 and TGF-${\beta}_1$ expression in lung lesions were examined using immunohistochemical staining. After $Bosentan^{(R)}$ administration, an orally active ET-$1_A$ and ET-$1_B$ receptor antagonist, the degree of pulmonary fibrosis and ET-1 and TGF-${\beta}_1$ expression were analyzed. Method : Sprague-Dawley rats were divided into three groups, the control group, the fibrosis group, and the fibrosis-$Bosentan^{(R)}$-treated group. The animals were sacrificed periodically at 1, 3, 5, 7, 10, 14 days after administering saline or paraquat. The effects between groups were compared with the results of light microscopy and immunohistochemical staining for ET-1 and TGF-${\beta}_1$. The degree of fibrosis was evaluated by H&E and Masson's trichrome staining, which were graded by a computerized image analyzer. The degree of immunohistochemical staining was categorized by a semi-quantitative analysis method. Results : The lung collagen content had increased in the paraquat instillated animals by day 3, and continued to increase up to day 14. A daily treatment by gavage with $Bosentan^{(R)}$ (100mg/kg) did not prevent the increase in collagen deposition on the lung that was induced by paraquat instillation. There were increased immunohistochemical stains of ET-1 on the exudate, macrophages, vascular endothelial cells and pneumocytes in the paraquat instillated group. Furthermore, TGF-${\beta}_1$ expression was higher on the exudate, macrophages, some inflammatory cells, pneumocytes( type I, and II), vascular endothelium and the respiratory epithelial cells around the fibrotic area. After Bosentan treatment, there were no definite changes in ET-1 and TGF-${\beta}_1$ expression. Conclusion : Fibrosis of the Paraquat instillated group was more advanced when compared with the control group. In addition, there was increased ET-1 and TGF-${\beta}_1$ expression around the fibrotic area. ET-1 is associated with lung fibrosis but there was little effect of the ET-1 receptor blocker($Bosentan^{(R)}$) on antifibrosis.

• Radiation Oncology Journal
• /
• v.19 no.4
• /
• pp.327-334
• /
• 2001

Purpose : To find out the role of postoperative adjuvant radiotherapy in the treatment of rectal cancer by comparing survival, pelvic control, complication rate, and any prognostic factor between surgery alone and postoperative radiotherapy group. Materials and methods : From Feb. 1982 to Dec. 1996 total 212 patients were treated by radical surgery with or without postoperative radiotherapy due to rectal carcinoma of modified Astler-Coiler stage $B2\~C3$. Of them, 18 patients had incomplete radiotherapy and so the remaining 194 patients were the database analyzed in this study. One hundred four patients received postoperative radiotherapy and the other 90 patients had surgery only. Radiotherapy was peformed in the range of $39.6\~55.8\;Gy$ (mean: 49.9 Gy) to the whole pelvis and if necessary, tumor bed was boosted by $5.4\~10\;Gy$. Both survival and pelvic control rates were calculated by Kaplan-Meier method and their statistical significance was tested by Log-rank test. Multivariate analysis was peformed by Cox proportional hazards model. Results : 5-year actuarial survival rate (5YSR) and 5-year disease-free survival rate (5YDFSR) of entire patients were $53\%\;and\;49\%$, respectively. 5YSRs of surgery alone group and adjuvant radiotherapy group were $63\%\;vs\;45\%$, respectively (p=0.03). This difference is thought to reflect uneven distribution of stages between two treatment groups (p<0.05 by $\chi^2-test$) with more advanced disease patients in adjuvant radiotherapy group. 5YSRs of surgery alone vs adjuvant radiotherapy group in MAC B2+3, C1, C2+3 were $68\%\;vs\;55\%$ (p=0.09), $100\%\;vs\;100\%$, $40\%\;vs\;33\%$ (p=0.71), respectively. 5YDFSRs of surgery alone vs adjuvant radiotherapy group in above three stages were $65\%\;vs\;49\%$ (p=0.14), $100\%\;vs\;100\%$, $33\%\;vs\;31\%$ (p=0.46), respectively. 5-year pelvic control rate (5YPCR) of entire patients was $72.5\%$. 5YPCRs of surgery alone and adjuvant radiotherapy group were $71\%\;vs\;74\%$, respectively (p=0.41). 5YPCRs of surgery alone vs adjuvant radiotherapy group in B2+3, C1, C2+3 were $79\%\;vs\;75\%$ (p=0.88), $100\%\;vs\;100\%$, $44\%\;vs\;68\%$ (p=0.01), respectively. Multivariate analysis showed that only stage was significant factor affecting overall and disease-free survival in entire patients and also in both treatment groups. In view of pelvic control, stage and operation type were significant in entire patients and only stage in surgery alone group but in adjuvant radiotherapy group, operation type instead of stage was the only significant factor in multivariate analysis as a negative prognostic factor in abdominoperineal resection cases. Conclusion : Our retrospective study showed that postoperative adjuvant radiotherapy could improve the pelvic control in MAC C2+3 group. To improve both pelvic control and survival in all patients with MAC B2 or more, other treatment modality such as concurrent continuous infusion of 5-FU, which is the most standard agent, with radiotherapy should be considered.

• Radiation Oncology Journal
• /
• v.17 no.2
• /
• pp.91-99
• /
• 1999

Propose : This study was performed to find out the prognostic factors affecting local control, survival and disease free survival rate in nasopharyngeal carcinomas treated with chemotherapy and radiation therapy. Materials and Methods : We analysed 47 patients of nasopharyngeal carcinomas, histologically confirmed and treated at Chonnam University Hospital between July 1986 and June 1996, retrospectively. Range of patients' age were from 16 to 80 years (median; 52 years). Thirty three (70$\%$) patients was male. Histological types were composed of 3 (6$\%$) keratinizing, 30 (64$\%$) nonkeratinizing squamous cell carcinoma and 13 (28$\%$) undifferentiated carcinoma. Histoiogicai type was not known in 1 patient (2$\%$). We restaged according to the staging system of 1997 American Joint Committee on Cancer Forty seven patients were recorded as follows: 71: 11 (23$\%$), T2a; 6 (13$\%$), T2b; 9 (19$\%$), 73; 7 (15$\%$), 74: 14 (30$\%$), and NO; 7 (15$\%$), Nl: 14 (30$\%$), N2; 21 (45%), N3: 5 (10%). Clinical staging was grouped as follows: Stage 1; 2 (4$\%$), IIA: 2 (4$\%$), IIB; 10 (21$\%$), III; 14 (30$\%$), IVA; 14 (30$\%$) and IVB; 5 (11$\%$). Radiation therapy was done using 6 MV and 10 MV X- ray of linear accelerator. Electron beam was used for the Iymph nodes of posterior neck after 4500 cGy. The range of total radiation dose delivered to the primary tumor was from 6120 to 7920 cGy (median; 7020 cGy). Neoadjuvant chemotherapy was performed with cisplatin +5-fluorouracil (25 patients) or cisplatin+pepleomycin (17 patients) with one to three cycles. Five patients did not received chemotherapy. Local control rate, survival and disease free suwival rate were calculated by Kaplan-Meier method. Generalized Wilcoxon test was used to evaluate the difference of survival rates between groups. multivariate analysis using Cox proportional hazard model was done for finding prognostic factors. Results: Local control rate was 81$\%$ in 5 year. Five year survival rate was 60$\%$ (median survival; 100 months). We included age, sex, cranial nerve deflicit, histologic type, stage group, chemotherapy, elapsed days between chemotherapy and radiotherapy, total radiation dose, period of radiotherapy as potential prognostic factors in multivariate analysis. As a result, cranial none deficit (P=0.004) had statistical significance in local control rate. Stage group and total radiation dose were significant prognostic factors in survival (P=0.000, P=0.012), and in disease free survival rates (P=0.003, P=0.008), respectively. Common complications were xerostomia, tooth and ear problems. Hypothyroidism was developed in 2 patients. Conclusion : In our study, cranial none deficit was a significant prognostic factor in local control rate, and stage group and total radiation dose were significant factors in both survival and disease free survival of nasopharyngeal carcinoma. We have concluded that chemotherapy and radiotherapy used in our patients were effective without any serious complication.

• Nuclear Medicine and Molecular Imaging
• /
• v.40 no.4
• /
• pp.218-227
• /
• 2006

Purpose: The HSV1-tk reporter gene system is the most widely used system because of its advantage that direct monitoring is possible without the introduction of a separate reporter gene in case of HSV1-tk suicide gene therapy. In this study, we investigate the usefulness of the reporter probe (substrate), $9-(4-[^{18}F]Fluoro-3-hydroxymethylbutyl)$guanine ($[^{18}F]FHBG$) for non-invasive reporter gene imaging using PET in HSV1-tk expressing hepatoma model. Materials and Methods: Radiolabeled FHBG was prepared in 8 steps from a commercially available triester. The labeling reaction was carried out by NCA nucleophilic substitution with $K[^{18}F]/K2.2.2.$ in acetonitrile using N2-monomethoxytrityl-9-14-(tosyl)-3-monomethoxytritylmethylbutyl]guanine as a precursor, followed by deprotection with 1 N HCl. Preliminary biological properties of the probe were evaluated with MCA cells and MCA-tk cells transduced with HSV1-tk reporter gene. In vitro uptake and release-out studies of $[^{18}F]FHBG$ were performed, and was analyzed correlation between $[^{18}F]FHBG$ uptake ratio according to increasing numeric count of MCA-tk cells and degree of gene expression. MicroPET scan image was obtained with MCA and MCA-tk tumor bearing Balb/c-nude mouse model. Results: $[^{18}F]FHBG$ was purified by reverse phase semi-HPLC system and collected at around 16-18 min. Radiothemical yield was about 20-25%) (corrected for decay), radiochemical purity was >95% and specific activity was around >55.5 $GBq/{\mu}\;mol$. Specific accumulation of $[^{18}F]FHBG$ was observed in HSV1-tk gene transduced MCA-tk cells but not in MCA cells, and consecutive 1 hour release-out results showed more than 86% of uptaked $[^{18}F]FHBG$ was retained inside of cells. The uptake of $[^{18}F]FHBG$ was showed a highly significant linear correlation ($R^2=0.995$) with increasing percentage of MCA-tk numeric cell count. In microPET scan images, remarkable difference of accumulation was observed for the two type of tumors. Conclusion: $[^{18}F]FHBG$ appears to be a useful as non-invasive PET imaging substrate in HSV1-tk expressing hepatoma model.

• Clinical and Experimental Pediatrics
• /
• v.50 no.11
• /
• pp.1097-1103
• /
• 2007

Purpose : Acute symptomatic seizure is defined as a temporary seizure together with acute systemic, metabolic, or toxic insult in association with an acute central nervous system insult. And unprovoked seizure is defined as seizure without provocating factors. We studied the risk factors of unprovoked seizures after acute symptomatic seizure in children. Methods : We retrospectively reviewed the records of one hundred and ten children with acute symptomatic seizures who were admitted to the pediatric department of Chungbuk National University Hospital between January, 1998 and December, 2003. We analyzed overall risk factors of unprovoked seizures after acute symptomatic seizures involving etiology, incidence, type of seizure, duration and neuroimaging. Results : We analyzed records of 110 children with acute symptomatic seizures aged from 1 month to 17 years. 24 children had unprovoked seizures (21.8%) after acute symptomatic seizures. Causes in order of frequency were encephalopathy, central nervous system infection, brain tumor, cerebrovascular disease. The risk of unprovoked seizure was significantly greater for those with status epilepticus (68.4%) than without status epilepticus, with partial seizure (64.7%) than generalized seizure. And the risk of unprovoked seizure was strongly associated with abnormal finding of electroencephalogram (79.1%) and neuroimaging (41.6%). Conclusion : In conclusion, the leading cause of subsequent unprovoked seizure in children with acute symptomatic seizure was encephalopathy and age specific incidence was high in the group aged 24-72 months. The risk for subsequent unprovoked seizure was greater for those with partial seizure, status epilepticus, abnormal finding of neuroimaging and electroencephalography.

• Journal of Oral Medicine and Pain
• /
• v.30 no.3
• /
• pp.287-300
• /
• 2005

Squamous cell carcinoma (SCC) in head and neck show a variability in the response to chemotherapy, even when it present with similar histological tumor type, grade, and clinical stage. The purpose of present study it to identify predictive bio-marker for the sensitivity or resistance to conventional chemotherapeutic agents, 5-fluorouracil (5-FU) and Cisplatin Oral cancer cell lines were used in present study. MTT assay was performed to evaluate the sensitivity and/or resistance to 5-FU and Cisplatin. And RT-PCR was carried out for evaluation of the mRNA expressions of various genes associated with mutation, inflammation (COX pathway), cell cycle, senescence and extracellular matrix (ECM). The molecules which are correlated with the sensitivity to 5-FU are XPA, XPC, OGG, APEX, COX-2, PPAR, Cyclin E, Cyclin B1, CDC2, hTERT, hTR, TIMP-3, TIMP-4 and HSP47. And the molecules are correlated with the sensitivity to Cisplatin are COX-1, iNOS, eNOS, PCNA, collagen 1 and MMP-9. Taken together, when choosing the appropriate chemotherpeutic agents for patients, considering the molecules which are correlated or reversely correlated is helpful to choose the resonable agents for cancer patients.

• Radiation Oncology Journal
• /
• v.13 no.1
• /
• pp.69-78
• /
• 1995

Seventy four patients with soft tissue sarcomas treated by postoperative radiotherapy in the Department of Therapeutic Radiology, Seoul National University Hospital between August 1979 and September 1990 were analyzed. The follow-up Period ranged from 3 to 145 months with a median of 51 months. Liposarcoma and malignant fibrous histiocytoma(MFH) constituted $54\%$ of the cases and the histologic grades of tumors are as follows: grade I, 23 cases; grade II, 17 cases; grade III, 24 cases: unknown grade, 10 cases. The patients were treated by marginal(17 cases), wide(55 cases) or compartmental(2 cases) excision followed by Postoperative radiotherapy. The total radiation doses were 4200-8820 cGy (median 6000 cGy), 180-200 cGy daily. 5 times per week. Of 74 Patients, 35 ultimately failed. The local control was $62.2\%$ at 5 years and cumulative risk of distant metastasis was $22,3\%$ at 5 years. The overall survival and disease free survival were $72.3\%$, and $53.3\%$ at 5 years, respectively. Survival after appearance of metastasis was $15.1\%$ at 3 years. Patients with liposarcoma experienced better local control than those with other histologic type and tumor grade and surgical resection margin significantly correlated with local recurrence, distant metastasis and overall survival on univariate analysis. In conclusion, re-excision is needed for patients with positive surgical resection margin to improve local control and further therapeutic measures using effective chemotherapy should be explored in the hope of improving overall survival.

• Tuberculosis and Respiratory Diseases
• /
• v.44 no.5
• /
• pp.1019-1029
• /
• 1997

Background : Elevation of resting energy expenditure(REE) in patients with lung cancer has been described in earlier studies and may contribute to cancer cachexia, but limited information is available regarding the prevalence and determinants of the increased REE. The aim of this study was to assess the prevalence and contributing factors of a hypermetabolic state in newly detected patients with lung cancer and to assess the energy balance in order to improve our knowledge about weight loss in patients with lung cancer. Method : Thirty one consecutive, newly detected patients with lung cancer and 20 control patients with benign lung diseases were included in this study. Resting energy expenditure(REE) was measured by indirect calorimetry using ventilated hood system and predicted REE was calculated by the Harris-Benedict formular. Results : The energy balance in newly detected lung cancer patients was disturbed in a high proportion of patients, and hypermetabolic state occurred in 61% of the patients. Tumor volume, cancer type, location, stage, the presence of atelectasis or infiltration, pulmonary function, or smoking behavior were not associated with increase in REE. But patients with distant metastasis had significantly higher REE comparing with patients without metastasis. Thirty nine percents of the patients with lung cancer had substantial loss of more than 10% of their pre-illness weight. Weight losing patients with lung cancer were not accompanied by an increase in REE. Conclusion : We concluded that the REE was elevated in a higher proportion of patients with lung cancer and distant metastasis was found to be contributing factor to the elevated REE.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.42 no.3
• /
• pp.335-341
• /
• 2013

Kyungokgos purchased in local markets in Korea vary in their combination and mixing ratios during processing. This study was investigated qualities of Kyungokgos manufactured traditionally to evaluating its qualities. The general components of Kyungokgos were moisture (18.62~49.78%), ash (0.198~1.211%), protein (0.89~3.58%), lipid (0.16~1.14%) and carbohydrates (47.95~77.08%). The color values of L, a, and b were 26.49~73.87, 16.51~38.64, and 45.41~88.94, respectively. The viscosity was classified into three non-Newtonian type groups: high, medium, and non-dilatant, according to the increase of loop execution times. Three extracts (KOG-1, -7, and -8, in a 30-fold dilution) showed no cytotoxicity toward RAW 264.7 cells, while the extracts of KOG-2, -4, and -5 showed a low cytotoxic effect. KOG-1 and -2 extracts with low cytotoxicity markedly inhibited the production of the inflammatory mediators-nitric oxide (NO) and tumor necrosis factor-alpha (TNF-${\alpha}$) in LPS-stimulated RAW 264.7 cells. These results indicate that KOG-1 and -2 extracts have anti-inflammatory activity in LPS-stimulated RAW 264.7 macrophages.