• Title/Summary/Keyword: Blood Assay

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Effects of Fermentation on the Metabolic Activities of Pine Needle Juice (발효과정이 솔잎 착즙액의 항산화, alpha-Glucosidase 및 Angiotensin Converting Enzyme 저해 활성에 미치는 영향)

  • Kim, So-Yun;Lee, Hyun-Jung;Park, Jae-Hee;Kim, Rae-Young;Cheong, Hyeonsook;Park, Eunju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.3
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    • pp.325-334
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    • 2013
  • The objective of this study was to compare the content and metabolic activities between fresh pine needle juice (PNJ) and fermented pine needle juice (FPNJ). A variety of factors were measured, including total phenolic content (TPC), antioxidant activity [DPPH radical scavenging activity (RSA), total radical-trapping antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), cellular antioxidant capacity (CAC)], anti-genotoxic activity, ${\alpha}$-glucosidase inhibitory activity, and angiotensin converting enzyme (ACE) inhibitory activity. The TPC was $17.3{\pm}0.2$ and $4.6{\pm}0.0$ mg GAE/g in PNJ and FPNJ, respectively. The DPPH RSA, TRAP, and ORAC values increased in a dose-dependent manner for both PNJ and FPNJ, with significantly higher activities in PNJ than FPNJ. The CAC against AAPH-induced oxidative stress in HepG2 cells was protected by both PNJ and FPNJ. Pretreatment with PNJ and FPNJ in human leukocytes produced significant reductions in $H_2O_2$-induced DNA damage at a concentration of $50{\mu}g/mL$. ${\alpha}$-Glucosidase inhibitory activity was significantly higher in FPNJ than PNJ. The ACE inhibitory activity was about 87.1% and 60.0% in 1:1 diluted PNJ and FPNJ, respectively. This study suggests that the fermentation of PNJ could enhance the regulation of blood glucose metabolism and both PNJ and FPNJ might be a new potential source of natural antioxidant, anti-diabetic, and anti-hypertensive agents applicable to food.

Estimation of Anti-proliferative Activity of Saccharin against Various Cancer Cell Lines and MSCs (다양한 암세포 주와 MSCs에 대한 Saccharin의 항증식성 평가)

  • Choi, Jeong Su;Park, Sang Yong;Yang, Man Gil;Lee, Dong Beom;Lee, Tae Bok;Heo, Ji Hye;Lee, Min Woo;Kim, Suhng Wook
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.169-175
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    • 2016
  • Saccharin (o-benzoic sulfimide) is the first artificial and non-caloric sweetener that was first synthesized in 1879. In this study, we examined the biological activity of saccharin against various human cancer cell lines and human bone marrow-derived mesenchymal stem cells. A viability assay based on the conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was performed to test for the cytotoxicity of saccharin about the four human cancer cell lines (H460, H157, A549 and SKOV3), one murine cancer cellline (Raw264.7), and MSCs. In order to find the differentially expressed gene in saccharin-treated MSCs against untreated MSCs, we performed annealing control primer (ACP)-based differential display reverse transcriptionp-olymerase chain reaction (DDRT-PCR). All tested cells were treated with saccharin at various concentrations (0.0, 4.8, 7.2, 9.6, 12.0, 14.4 mg/mL) for 48 hr. The number of metabolically active cancer cells decreased when treated with the saccharin at various concentrations for 48 hr as compared with the untreated cells. The decrease in cell survival was more evident with increasing concentrations of saccharin. Moreover, novel candidate genes, which were differentially expressed in MSCs in response to saccharin, were identified in 16 bands on 2% agarose gel. This revealed 16-7 up-regulated and 9 down-regulated-differentially expressed genes indicated by arrows. One of these candidate genes was a FK506-binding protein gene. The functional roles of FK506 binding proteins, with respect to the activities of stem cell proliferation, were not characterized. Further studies are required to get a better understanding of FK506-binding proteins in its roles in increasing stem cell proliferative activities from using saccharin.

Effect of Cosmetics Contained Isotonic Water Mimicked Body Fluid on Cell Activities and Skin (생체 모사수 화장품이 세포 활성과 피부에 미치는 효과)

  • Park, Sun Young;Lee, Sung Hoon;Kim, Eun Joo;Choi, So Woong;Kim, Ji Young;Cho, Seong A;Cho, Jun Cheol;Lee, Hae Kwang
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.2
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    • pp.195-201
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    • 2014
  • Body fluid has been studied for diverse fields like Ringer's solutions, artificial joint fluids, cell growth culture media because it plays a crucial role in controlling body temperature and acts as a solvent for diverse metabolite processes in the body and delivery media of mineral, energy source, hormone, signal and drug from and to cell via blood or lymphatic vessel by osmotic pressure or active uptake. Stratum corneum containing extracellular lipids and NMF (natural moisturizing factor) absorbs atmospheric water residing outside of cells and utilize it to hydrate inside of their own. This process is related to skin barrier function. In this study, we conducted the cell viability test with Cell Bio Fluid $Sync^{TM}$, which mimicks body fluids including amino acids, peptides, and monosaccharides to strengthen skin barrier, and the clinical skin improvement test with cosmetics containing Cell Bio Fluid $Sync^{TM}$. In the cell viability test, HaCaT cell was treated with PBS for 3 hours, followed by the treatment of a cell culture medium (DMEM) and isotonic solution (PBS) and Cell Bio Fluid $Sync^{TM}$ for 3 hours each. Then, MTT assay and image analysis were conducted. In the clinical skin improvement test, twenty-one healthy women participated. Participants applied cosmetics containing Cell Bio Fluid $Sync^{TM}$ on their face for a week and evaluated the skin hydration, skin roughness, brightness and evenness. All measurements were conducted after they washed off their face and took a rest under the constant temperature ($22{\pm}2^{\circ}C$) and constant humidity conditions ($50{\pm}5%$) for 20 minutes. All the data were analyzed by SPSS (version 21) software program. Results showed that Cell Bio Fluid $Sync^{TM}$ improved both the cell viability and in vivo skin conditions such as skin hydration, roughness, brightness and evenness.

Embryotoxicity and Teratogenicity of Excess Zinc on Xenopus laevis (과량의 아연에 의한 아프리카 발톱개구리 (Xenopus laevis)의 배발생 이상과 독성)

  • Yoon, Chun-Sik;Jin, Jung-Hyo;Cheong, Seon-Woo
    • Korean Journal of Ecology and Environment
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    • v.36 no.1 s.102
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    • pp.83-94
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    • 2003
  • Concentrated releases of zinc into water usually results from discharges associated with industrial purpose. The released zinc into soil is corroded and released into water. In aquatic environment, exess zinc is toxic to the organisms and causes the growth inhibition and malformation of them as a heavy metal. In this study, excess zinc toxicity was tested by FETAX (frog embryo teratogenetic assay with Xenopus)as in vivo system. Xenopus embryos at st.9 were exposed to $100{\sim}900\;{\mu}M$ of zinc for 7 days and 81% of individuals were survived in 100 ${\mu}M$, and 25% were survived in 1000M of zinc solution. In external malformations, swelled belly and intestinal dysplasia were common, and all of tested individuals showed these malformations in 200 ${\mu}M$ or higher concentration of zinc. In 400 ${\mu}M$ or higher concentration, all of tested tadpoles showed faded heart. Also, hypo-pigmentation, lens hernia and loose digestive track were very frequently found in 100 ${\mu}M$ of zinc. The histological study with paraffin section of zinc treated tadpoles showed following abnormalities; regeneration of photoreceptor on retina, reduced vitreous chamber in eye, reduction of red blood cells in heart, abnormal liver, swelling of pronephric cell, muscle dysplasia and palatal papilloma. These abnormalities may be caused by the degeneration of mitochondria, inhibition of cell adhesion, and the formation of leghemoglobin by zinc due to the substitution of $Ca^{2+}$ by $Zn^{2+}$. The body length was reduced due to the excess zinc. From a statistical result, body lengths of 300 ${\mu}M$ or higher concentrative g개ups was significantly reduced comparing that of control group. Recently, many spontaneous malformations and reduction of amphibians are reported, From the results of present study, excess zinc mi호t be a factor of amphibian reduction, and the control of zinc discharges is very important.

Comparison of Stress Response between Korean Native Chickens and Single Comb White Leghorns subjected to a High Stocking Density (닭의 품종 간 스트레스 반응 정도 비교 분석)

  • Sohn, Sea Hwan;Cho, Eun Jung;Park, Dhan Bee;Jang, In Surk;Moon, Yang Soo
    • Korean Journal of Poultry Science
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    • v.41 no.2
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    • pp.115-125
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    • 2014
  • With Single Comb White Leghorn (WL) and Korean Native Chicken (KNC) breeds, we compared the stress response with chicken breeds that were subjected to a high stocking density. Stress response was analyzed by the quantity of telomeric DNA, the rate of DNA damage and the expression levels of heat shock proteins (HSPs) and hydroxyl-3-methyl-glutaryl coenzyme A reductase (HMGCR) genes on tissues and blood. The telomere length and telomere shortening rates were analyzed by quantitative fluorescence in situ hybridization on the nuclei of lymphocytes and tissues. The DNA damage rate of lymphocytes was quantified by the comet assay. The expression levels of HSP70, HSP90-${\alpha}$, HSP90-${\beta}$ and HMGCR genes were measured by quantitative real-time polymerase chain reaction in lymphocytes. There was no significant difference between KNC and WL in body weight, weight gain, telomere shortening rate and DNA damage rate. However, the growth rate significantly decreased in chickens raised under high stocking density conditions, as compared to the control group. The telomere-shortening rate, DNA damage and HSPs expression of the lymphocytes were significantly higher in the high stocking density group than the control. The stress condition and breeds had a significant effect on the expressions of HSP70, HSP90-${\alpha}$ and HSP90-${\beta}$ in lymphocytes, except HMGCR. The stress response of WL was higher than that of KNC, as analyzed to the expression of HSP70 and HSP90-${\alpha}$. Therefore, we concluded that the chickens which were exposed to a high stocking density had increased the individual physiological stress response regardless of breeds, and White Leghorns are more susceptible to stress condition than Korean Native Chickens.

Usefulness of Interferon-$\gamma$ Measurement Following Stimulation of Tuberculosis-specific Antigens for Diagnosis of Latent Tuberculosis Infection in Children Exposed to Pulmonary Tuberculosis (폐결핵 접촉력을 가진 소아에서 잠복 결핵 감염의 진단을 위한 결핵 특이항원 자극 인터페론-감마 측정의 유용성)

  • Sung, Ji Yeon;Ki, Joo Hwa;Yang, Mi Ae;Kim, So Hee;Eun, Byung Wook;Kim, Nam Hee;Park, Kyoung Un;Lee, Jina;Choi, Eun Hwa;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.15 no.1
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    • pp.52-58
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    • 2008
  • Purpose : Whole blood interferon-$\gamma$ assay was developed and many studies showed its usefulness in diagnosing tuberculosis (TB) including latent tuberculosis infection (LTBI). However, assessment in children has been limited. This study was undertaken to evaluate the usefulness of QuantiFERON-TB Gold for the diagnosis of LTBI in children exposed to pulmonary TB. Methods : Children who visited Seoul National University Bundang Hospital with a history of TB exposure were enrolled from January 2006 to December 2007. They were evaluated with chest x-rays, tuberculin skin test (TST) and QuantiFERON-TB Gold test. TST was retested 3 months later for those with initial negative reactivity. Definition of LTBI was made on the basis of the TST reactivity. Results : Among the 103 children with a history of TB exposure, 49 children were tested with chest x-ray, TST, and QuantiFERON-TB Gold. Twenty-two were males. Median age was 7.5 years (range; 3 months to 14.7 years). According to TST reactivity, LTBI was in 8 (19%), no infection was in 21 (50%), possible LTBI was in 13 (31%). QuantiFERON-TB Gold test was positive in 5 of the 49 subjects (10%); 3 of the 13 subjects (23.1%) in unknown status, 1 of the 8 subjects (13%) in LTBI, and 1 of the 21 subjects (5%) without infection. The agreement between the QuantiFERON-TB Gold and the TST was poor (${\kappa}=0.101$). Conclusion : QuantiFERON-TB Gold showed poor sensitivity for the diagnosis of LTBI in children with exposure to TB. QuantiFERON-TB Gold alone does not seem to be useful in the diagnosis of LTBI in children.

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Prevalence of Feline Leukemia Virus Infection in Cats in Bangladesh (방글라데시의 고양이 백혈병 바이러스의 감염율 조사)

  • Rahman, Siddiqur;Bhuiyan, Salauddin;Islam, Taohidul;Nahar, Azimun;Sarker, Roma Rani;Alam, Emtiaj;Chakrabarty, Amitavo;Sarker, Abu Sayed;Akhter, Laila;Chae, Joon-Seok
    • Journal of Veterinary Clinics
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    • v.31 no.1
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    • pp.1-5
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    • 2014
  • Feline leukemia virus (FeLV) is a retrovirus that represents one of the most common and important infectious diseases of cats worldwide and it is responsible for more deaths among cats than any other infectious diseases. Prevalence data are necessary to define prophylactic, management and therapeutic measures for stray, feral and owned cats which are lacking in Bangladesh. The study was carried out to determine the prevalence of FeLV infection in Mymensingh district of Bangladesh using RapiGEN$^{(R)}$ FeLV Ag Test Kit (RapiGEN$^{(R)}$ Inc., Republic of Korea), a rapid one-step immunochromatographic assay. Blood samples from total 130 cats (23 owned cats and 107 unowned cats) were collected and tested following the manufacturer's instruction. An overall prevalence of FeLV infection was 1.54% (2/130). Prevalence was found 1.79% (2/112) on Day 0-up to one year aged cats (young) but no positive case was found in above 1 year (Adult) aged group. In male and female cats, the prevalence was 1.72% (1/58) and 1.39% (1/72), respectively. In un-owned cats the prevalence was 1.87%. Positive cases to FeLV were found only in clinically sick cats. No significant relationship was found according to age, sex, ownership status and health status. To the best of our knowledge this is the first report of the prevalence of FeLV infection in Bangladesh using RapiGEN$^{(R)}$ FeLV test kits which is very much effective because it is easy to apply, less expensive and quick screening of such type of infection.

Role of p-38 MAP Kinase in apoptosis of hypoxia-induced osteoblasts (저산소 상태로 인한 조골세포 고사사기전에서 p-38 MAP kinase의 역할에 관한 연구)

  • Yoon, Jeong-Hyeon;Jeong, Ae-Jin;Kang, Kyung-Hwa;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.33 no.3 s.98
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    • pp.169-183
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    • 2003
  • Tooth movement by orthodontic force effects great tissue changes within the periodontium, especially by shifting the blood flow in the pressure side and resulting in a hypoxic state of low oxygen tension. The aim of this study is to elucidate the possible mechanism of apoptosis in response to hypoxia in MC3T3El osteoblasts, the main cells in bone remodeling during orthodontic tooth movement. MC3T3El osteoblasts under hypoxic conditions ($2\%$ orygen) resulted in apoptosis in a time-dependent manner as estimated by DNA fragmentation assay and nuclear morphology stained with fluorescent dye, Hoechst 33258. Pretreatment with Z-VAD-FMK, a pancaspase inhibitor, or Z-DEVD-CHO, a specific caspase-3 inhibitor, completely suppressed the DNA ladder in response to hypoxia. An increase in caspase-3-like protease (DEVDase) activity was observed during apoptosis, but no caspase-1 activity (YVADase) was detected. To confirm what caspases are involved in apoptosis, Western blot analysis was performed using anti-caspase-3 or -6 antibodies. The 10-kDa protein, corresponding to the active products of caspase-3, and the 10-kDa protein of the active protein of caspase-6 were generated in hypoxia-challenged cells in which the processing of the full length form of caspase-3 and -6 was evident. While a time course similar to this caspase-3 and -6 activation was evident, hypoxic stress caused the cleavage of lamin A, which was typical of caspase-6 activity. In addition, the stress elicited the release of cytochrome c into the cytosol during apoptosis. Furthermore, we observed that pre-treatment with SB203580, a selective p38 mitogen activated protein kinase inhibitor, attenuated the hypoxia-induced apoptosis. The addition of SB203S80 suppressed caspase-3 and -6-like protease activity by hypoxia up to $50\%$. In contrast, PD98059 had no effect on the hypoxia-induced apoptosis. To confirm the involvement of MAP kinase, JNK/SAPK, ERK, or p38 kinase assay was performed. Although p38 MAPK was activated in response to hypoxic treatment, the other MAPK -JNK/SAPK or ERK- was either only modestly activated or not at all. These results suggest that p38 MAPK is involved in hypoxia-induced apoptosis in MC3T3El osteoblasts.

A Study on The Content of Liver Protein, Nucleic Acids, and Guanine Deaminase Activity of Mouse During Acute Starvation (급성(急性) 기아(饑餓)마우스의 간단백질(肝蛋白質), 핵산(核酸) 및 Guanine Deaminase 활성(活性)에 관(關)한 연구(硏究))

  • Park, Seung-Hee;Kim, Seung-Won
    • Journal of Nutrition and Health
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    • v.1 no.2
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    • pp.107-115
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    • 1968
  • Number of aspects, not only nutritional but social as well as political involved in human starvation pose nowadays global problems. In order to help establish the minimum nutritional requirements in the daily life of a man and to free people as well from either undernourishment, malnutrition or even starvation many workers have devoted themselves so far on the research programs to know what and how number of metabolic events take place in animals in vivo. It is the purpose of the present paper to examine in effect to what extent both of the protein and nucleic acids (DNA & RNA) together with an enzyme, guanine deaminase, which converts guanine into xanthine and in turn ends up to uric acid as an end product, undergo changes, quantitatively during acute starvation, using the mouse as an experimental animal. The mouse was strictly inhibited from taking foods except drinking water ad libitum and was sacriflced 24, 48, and 72 hours following starvation thus acutely induced. The animals consisted of two experimental groups, one control and another starvation groups, each being consisted of 6-24 mice of whose body weights ranged in the vicinity of 10 g. The animals were sacriflced by a blow on the head, followed by immediate excision of their livers into ice-cold distilled water, washing adherent blood and other contaminant tissues. The liver was minced foramin, by an all-glass homogenizer immersing it in an ice-bath, followed by subsequent fractionatin of the homogenate (10% W/V in 0.25M sucrose solution made up with 0.05M phosphate buffer of pH 7.4). For the liver protein and guanine deaminase assay, the 10% homogenate was centrifuged at 600 x g for 10 minutes to eliminate the nuclear fraction; and for the estimation of DNA and RNA, the homogenate was prepared by the addition of 10% trichloroacetic acid in order to free the homogenate from the acid-soluble fraction, the remaining residue being delipidate by the addition of alcohol and dried in vacuo for later KOH (IN) hydrolysis. The changes in body and liver wegihts during acute starvation were checked gravimetrically. Protein contents in the liver were monitored by the method of Lowry et al; and guanine deaminase activities were followed by the assay of liberated ammonia from the substrate utilizing the Caraway's colorimetry. The extraction of both DNA and RNA was performed by the Schmidt-Thannhauser's method, which was followed by Marmur's method of purification for DNA and by Chargaff's method of purification for RNA. The determinations of both DNA and RNA were carried out by the diphenylamine reaction for the former and by the orcinol reaction for the latter. The following resume was the results of the present work. 1. It was observed that the body as well as liver weights fall abruptly during starvation, and that the loss of body weight showed no statistical correlation with the decreases in the content of liver protein. 2. The content of liver protein and activity of liver guanine deaminase activity as well decline dramatically, and the specific activities of the enzyme (activity/protein), however, decreased gradually as starvation proceeded. 3. Both of the nucleic acids, DNA and RNA, showed decrements in the liver of mouse during acute starvation; the latter, however, being more striking in the decline as compared to the former. 4. The decreases in the liver protein content as resulted from the acute starvation had no statistically significant correlation with the decrements of DNA in the same tissue, but had regressed with a significant statistical correlation with the fall of RNA in the tissue. 5. The decrease in the activity of guanine deaminase in the liver of mouse during acute starvation was functionally more proportional to the decrease in RNA than DNA, and moreover correlated with the changes in the content of the liver protein. 6. The possible mechanisms involved during in this acute starvation as bring the decreases in the contents of DNA, protein, and guanine deaminase were discussed briefly.

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Urine endothelin-1 as a Biomarker for Urinary Tract Infections in Children (Urine endothelin-1의 소아 요로 감염 환아에서의 진단적 유용성)

  • Kee, Hyung Min;Yi, Dae Yong;Yun, Ki Wook;Lim, In Seok;Ha, Tae-Seon
    • Childhood Kidney Diseases
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    • v.18 no.1
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    • pp.36-41
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    • 2014
  • Purpose: Urinary tract infections (UTIs) are the most common source of bacterial infections in infants and young children. Accurate diagnosis and treatment is important because of their association with renal scarring, which can lead to complications. Urine endothelin-1 (ET-1) is the major renal isoform produced and released by renal mesangial cells in response to glomerular injury. This study aimed to investigate whether urinary levels of ET-1 can be used as a biomarker for UTI diagnosis. Method: We conducted a prospective study using medical records of 70 patients below the age of 18 years, who visited Chung-Ang University Hospital from July 2012 to July 2013. We classified the patients into the UTI and control groups based on urine culture studies. The UTI group was further divided into upper and lower UTI groups using 99m-Technetium dimercaptosuccinic acid scintigraphy. Urine ET-1 was measured using enzyme linked immunosorbent assay with 0.3 mL urine. Results: The UTI and control groups were comprised of 45 and 25 patients, respectively. Mean urine ET-1 levels were significantly higher in the UTI group than in the control group ($1.41{\pm}0.35$ pg/mL vs. $0.33{\pm}0.07$ pg/mL, P =0.04). There was no significance difference in the quantitative value between the upper and lower UTI groups (P =0.552). There was no correlation between urine ET-1 and serum C-reactive protein (Pearson correlation [R]=0.24), urine ET-1 and serum white blood cell count (R=0.19). Conclusion: Our study suggests that urine ET-1 can be used for early diagnosis of UTI in children.