• Title/Summary/Keyword: ELISA method

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Macrophage Migration Inhibitory Factor (MIF) Concentration in the Serum of Patients with Ovarian Cysts for Differential Diagnosis of Endometrioma (자궁내막종의 감별진단을 위한 난소낭종 환자의 혈청 Macrophage Migration Inhibitory Factor (MIF) 농도의 진단적 유용성에 대한 연구)

  • Kim, Man Ki;Kim, Yu Re;Hong, Seong Hwa;Park, Yeon Jin;Ji, IL Woon;Jeong, Eun Hwan;Kim, Hak Soon
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.3
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    • pp.287-293
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    • 2005
  • Objective: To evaluate the usefulness of serum concentrations of macrophage migration inhibitory factor (MIF) of patients with ovarian cysts for differential diagnosis of endometrioama. Method: From Jan. 2003 to Dec. 2004, preoperative serum MIF levels were assessed in 28 women with endometrioma, 32 with benign epithelial tumor, 23 with functional and simple cysts, 22 with benign mature cystic teratoma, and 25 women without ovarian tumor as control. MIF levels were determined using an ELISA (Quantikine Human MIF immunoassay, R&D Systems, Inc., USA). Results: Mean MIF levels were higher in all groups with benign tumors than control (all p<0.01), but there was no significant difference between benign tumor groups (p=0.95). There was no significant correlation between MIF levels and tumor volume, body mass index (BMI) (p=0.635, 0.674 respectively) Serum MIF level had significant correlation with count of WBC and neutrophils (p=0.008, 0.024 respectively), but had no correlation with count of lymhocytes and monocytes (p=0.688, 0.294 respectively). Conclusions: This study showed a marked increase in MIF concentrations in the peripheral blood of patients with endometrioma, but there was no significant difference with other benign tumors. Serum MIF level had significant correlation with count of WBC and neutrophils. These suggest serum MIF level has no usefulness for differential diagnosis of endometrioma from other benign ovarian cysts.

Effect of Sihogayonggolmoryeotang on SPS-induced PTSD in Rats (시호가룡골모려탕(柴胡加龍骨牡蠣湯)이 흰쥐에서 SPS로 유도된 PTSD에 미치는 효과)

  • Kim, Hwi-Yeol;Lee, Tae Hee
    • Herbal Formula Science
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    • v.27 no.2
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    • pp.121-136
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    • 2019
  • Objective : To investigate the effect of sihogayonggolmoryeotang (SY) on Single Prolonged Stress(SPS)-induced Post Traumatic Stress Disorder(PTSD). Method : To confirm the effects of SY on SPS-induced PTSD, Changes in body weight, sucrose intake open field test(OFT) and forced swimming test(FST)were observed. After behavioral tests, the plasma corticosterone(CORT) from the abdominal aorta, serotonin(5-HT) from prefrontal cortex, hippocampus, amygdala and striatum, norepinephrine(NE) and dopamine(DA) from hippocampus was measured by ELISA. mRNA expression of brain-derived neurotrophic factor(BDNF) and cAMP response element-binding protein(CREB) in hippocampus was measured by RT-PCR. Result : Weight change and sucrose intakes of rats in 14th day after the administration of SY were significantly increased in the SPS + SY450 group compared to the SPS group (p<0.05). Numbers of crossing in the central zone in the OFT were significantly increased in the SPS + SY450 group (p<0.05) compared with the SPS group. The immobility time of FST was significantly decreased in SPS + SY450 group compared with SPS group (p<0.05). The change of plasma CORT concentration was significantly decreased in SPS + SY450 group compared with that in SPS group (p<0.05). The change of 5-HT concentration was significantly increased in the SPS + SY450 group at hippocampus and amygdala compared with the SPS group (p<0.05). The concentration of DA was significantly increased in the SPS + SY450 group compared with the SPS group (p<0.05). The expression of BDNF and CREB were significantly increased in SPS + SY450 group compared with the SPS group (p<0.05). Conclusion : SY administration lowered the increase of CORT caused by PTSD and increases the 5-HT concentration and reversed the decreased expression of NE and DA and BDNF and CREB by PTSD. It is postulated that SY is effective in treating PTSD by restoring cognitive function, memory impairment, unstable emotional disturbances.

Effect of Banggibongnyeongtang on LPS-induced Depression in rats (방기복령탕(防己茯苓湯)이 흰쥐에서 LPS로 유도된 우울증에 미치는 효과)

  • Park, Sung jun;Lee, Tae Hee
    • Herbal Formula Science
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    • v.27 no.2
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    • pp.137-149
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    • 2019
  • Objective : This study is conducted in order to investigate the effect of Banggibongnyeongtang(BBT) on Lipopolysaccharide (LPS)-induced depression. Method : LPS $5{\mu}g$ was injected to lateral ventricle. Experimental groups were administered BBT intraperitoneally. Depressive behavior was confirmed by weight change, sucrose preference, open field test(OFT), and forced swimming test(FST). The plasma concentration of $IL-1{\beta}$ and $TNF-{\alpha}$, Corticotropin-Releasing Factor(CRF), Adrenocorticotropin Hormone(ACTH) and Corticosterone(CORT) were measured by ELISA. Result : BBT did not change the body weight significantly than LPS group, but on sucrose preference, BBT increased significantly in LPS+BBT400 group compared to LPS group (P<0.05). In the OFT, BBT increased spending time in the central zone and decreased grooming number. LPS+BBT400 group increased central zone-spending time, and decreased grooming number than LPS group significantly (P<0.05). In the FST, LPS+BBT400 group decreased immobility time than LPS group significantly (P<0.05). BBT decreased $IL-1{\beta}$ concentration does-dependently, but only with significant decrease in LPS+BBT400 group than LPS group in plasma (P<0.05). But BBT did not decrease $TNF-{\alpha}$ concentration significantly in plasma. BBT decreased plasma CRF, ACTH, and CORT. And CRH and CORT of LPS+BBT400 group were shown significant decrease comparing with LPS group (P<0.05). Conclusion : It is postulated that the anti-depressant effect of BBT can be validated through inhibition of HPA axis abnormal activity by the anti-inflammatory effect.

Quantification of the Bioactive Components of the Rhizomes of Curcuma wenyujin and Assessment of Its Anti-inflammatory Effect in Benign Prostatic Hyperplasia-1 Cells

  • Seo, Chang-Seob;Park, Eunsook;Lee, Mee-Young;Shin, Hyeun-Kyoo
    • Natural Product Sciences
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    • v.25 no.3
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    • pp.238-243
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    • 2019
  • In this study, the marker compounds of Curcumae Rhizoma (CR) were simultaneously quantified by high-performance liquid chromatography equipped with a photodiode array detector and the anti-inflammatory effects of CR extract and marker compounds in human benign prostatic hyperplasia epithelial-1 (BPH-1) cell lines were investigated. The marker components (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, were separated on Gemini $C_{18}$ columns ($250mm{\times}4.6mm$, $5{\mu}m$) at $40^{\circ}C$ by using a gradient of two mobile phases eluting at 1.0 mL/min. Prostaglandin $E_2$ ($PGE_2$) levels in Human BPH-1 cells were determined with an ELISA kit. The coefficients of determination in a calibration curve of each analyte were all 0.9997. The limits of detection and quantification of the three compounds were $0.10-0.32{\mu}g/mL$ and $0.30-0.98{\mu}g/mL$, respectively. The content of three compounds, (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, in the CR sample were found to be 5.79 - 5.92 mg/g, 4.72 - 4.86 mg/g, and 1.06 - 1.09 mg/g, respectively. Regarding pharmacological activity against benign prostatic hyperplasia, CR and its components significantly suppressed $PGE_2$ levels of BPH-1 cells. The established analysis method will help to improve quality assessment of CR samples and related products. In addition, CR and its components exhibit antiinflammatory activity in BPH-1 cells, suggesting the inhibitory efficacy of these compounds against the pathogenesis of BPH.

Effect of vitamin C on pregnancy rate and 8-OHdG levels during heat stress in post-partum dairy cattle

  • Kirdeci, Armagan;Cetin, Hayrettin;Raza, Sanan
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.4
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    • pp.194-202
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    • 2021
  • In this study the effect of vitamin C administration on pregnancy rates during summer heat stress in dairy cows was examined. A total of 80 Holstein-Friesian cows were divided into control and treatment groups (n = 40 each). Control group animals were given 10 mL isotonic normal saline, and treatment group, Vitamin C (4 mg/kg) on artificial insemination day (day 0) and 4th, 8th and 12th day post insemination. Pregnancy diagnosis was performed on 30th day post insemination by ultrasonography. Blood samples were randomly taken from 11 animals from each group. Serum P4, GSH, MDA and plasma 8-OHdG levels were determined by using ELISA method. Results showed that 8-OHdG levels were lower in treatment group on day 4, 8 and 12 (p < 0.05) compared with the control group. Similarly, pregnancy rate was higher in treatment group (32.5%) than control (22.5%), respectively. However, MDA, P4 and GSH levels were similar in both groups at 4th, 8th and 12th day. A gradual increase in P4, and MDA levels, and a strong positive correlation between 0, 4th (r = 0.54), 4, 8th (r = 0.59) and 8, 12th (r = 0.51) day was found. Similarly, GSH levels also showed positive correlation at days 0, 4th (r = 0.47) and 4, 8th (r = 0.56). However, a strong negative correlation (r = -0.56) between MDA day 0, and GSH day 8 was found. In conclusion, vitamin C application during insemination period in postpartum cows increases pregnancy rate, and reduces oxidative stress metabolite 8-OHdG levels.

Effect of Glycyrrhizae Radix Water Extract on the depression induced by LPS in Rats (LPS로 유발된 우울증에 대한 감초(甘草)의 효과)

  • Ko, Sung-Youl;Kim, Do-Hoon;Lee, Tae-Hee
    • The Korea Journal of Herbology
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    • v.29 no.5
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    • pp.9-16
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    • 2014
  • Objectives : Investigation of the antidepressant effect of Glycyrrhizae Radix (GR) through the anti-inflammatory effect. Methods : Depression in rats was induced by LPS (i,p.3days). The rats were treated with GR100 mg/kg (GR 100) or GR400 mg/kg (GR 400). The depressive immobility was examined with Tail Suspension Test(TST) and Forced Swimming Test(FST). The expression of nuclear factor-${\kappa}B$(NF-${\kappa}B)$, $I{\kappa}B$ was measured with western blotting. The concentration of corticosterone, cytokine in plasma was measured with ELISA. The expression of c-Fos in the paraventricular nucleus(PVN) and tyrosine hydroxylase(TH) in the locus coeluleus(LC) were measured with immunostaining method. Results : In the TST, GR400 group significantly decreased immobility time compared with the LPS group. In the FST, GR100, GR400 group significantly decreased immobility time comparing with the LPS group. c-Fos expression in GR100 and GR400 group was decreased comparing with the lipoplysaccharide(LPS) group. The $I{\kappa}B$ expression of GR100 and GR400 group was increased comparing with the LPS group. The level of corticosterone of GR100 group was decreased comparing with the LPS group. The concentration of cytokine of GR100 and GR400 group was decreased comparing with the LPS group. TH expression in the LC was increased in LPS group, but in GR100 and GR400 group was not shown significant decrease. Conclusion : According to this results obtained, GR has antidepressant effects by the anti inflammatory action through the suppression of HPA axis activity, not through the action against the catecholaminergic system.

The Study of Cytokine Inhibitory Effect using Careswell (케어스웰을 이용한 사이토카인 억제 효과에 대한 연구)

  • Ha, Hyocheol;Oh, Sarang;Lee, Jihye;Jung, Ji Wook
    • The Korea Journal of Herbology
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    • v.29 no.3
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    • pp.79-85
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    • 2014
  • Objectives : In this study, we attempted to evaluate the effects of Careswell on human mast cell-mediated allergy inflammation in vitro and pruritogen-induced scratching behavior in vivo. Method : The Careswell was extract by distilled water. The anti-itching effects of Careswell were investigated on the compound 48/80 ($50{\mu}g/kg$) or histamine ($100{\mu}g/kg$) induced scratching behavior male ICR mice for 30 min by an observer blind. Terfenadine (10 mg/kg) was used as a positive control drug. The cell toxicity of Careswell was determined by 3-(4,5-dimethylthiazole-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. The regulatory effect of Careswell on interleukin (IL)-6 and tumor necrosis factor (TNF)-${\alpha}$ levels was determined by enzyme-linked immunosorbent assay (ELISA) in phorbol 12-myristate 13-acetate plus calcium ionophore A23187 (PMACI) stimulated human mast cells (HMC-1). Also, we evaluated the effect of Careswell on PMACI induced the activation of Nuclear factor-kappa B (NF-${\kappa}B$) into nucleus by Western blot analysis. Result : The results revealed that the oral administration of Careswell (200 mg/kg, p.o.) attenuated the compound 48/80 or histamine-induced scratching behavior in mice. We showed that Careswell significantly reduced the PMACI-induced the production of IL-6 (0.5-1 mg/ml) and TNF-${\alpha}$ (0.1-1 mg/ml). Additionally, Careswell significantly inhibited the activation of NF-${\kappa}B$ in PMACI-stimulated HMC-1. Conclusion : Collectively, the findings of this study provide us with a novel insight into the pharmacological actions of Careswell as a potential molecule for use in the treatment of allergic inflammation diseases.

Effects of Polygonati Rhizoma Extracts on the Collagenase Activity and Procollagen Synthesis in Hs68 Human Fibroblasts and Tyrosinase Activity

  • Park, Dong-Su;Shin, Seon-Mi;Leem, Kang-Hyun
    • The Korea Journal of Herbology
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    • v.28 no.3
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    • pp.1-5
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    • 2013
  • Objectives : This study was designed to investigate the collagen metabolism and tyrosinase activity of Polygonati Rhizoma extracts (PR). It's effects are to tonify spleen qi and augment the spleen yin. It enrichs the yin and moisten the lung. Methods : The effect of PR on type I procollagen production and collagenase (matrix metalloproteinase-1, henceforth referred as MMP-1) activity in human normal fibroblasts Hs68 after ultraviolet B (UVB, 312 nm) irradiation was measured by ELISA method. The tyrosinase activity after treatment of PR was measured. Results : There were no cytotoxicity at concentrations of 10, 30, $100{\mu}g/ml$. The reduced type I procollagen production was recovered by PR in UVB damaged Hs68 cells at a concentration of $100{\mu}g/ml$ ($16.2{\pm}0.0$ ng/ml) from control group ($13.9{\pm}0.5$ ng/ml). However there was no statistical significance. PR reduced The increased MMP-1 activity after UVB damage at concentrations of $10{\mu}g/ml$, $30{\mu}g/ml$, and $100{\mu}g/ml$ in a dose dependent manner ($42.2{\pm}20.5%$, $44.8{\pm}8.5%$, and $22.0{\pm}5.8%$). PR $100{\mu}g/ml$ treatment showed the statistical significace (p < 0.05). PR significantly reduced the tyrosinase activity at a concentration of 10 mg/ml ($32.0{\pm}12.8%$, p < 0.05). However, the L-DOPA oxidation was not changed. Conclusion : PR showed the anti-wrinkle effects and whitening effects in vitro. Although more researches are needed to validate the efficacy, these results suggest that PR may have potential as an anti-aging ingredient in cosmetic herb markets.

Effects of Bletillae Rhizoma on the Elastase, Collagenase, and Tyrosinase Activities and the Procollagen Synthesis in Hs68 Human Fibroblasts

  • Lee, Jung-Hun;Kim, Myung-Gyou;Lee, Sena;Leem, Kang-Hyun
    • The Korea Journal of Herbology
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    • v.28 no.1
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    • pp.9-14
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    • 2013
  • Objectives: Bletillae Rhizoma, the roots of Bletilla striata, is used to restrain the leakage of blood and stop bleeding. It can cure the sores, ulcers, and chapped skin. This study was designed to investigate the collagen metabolism, elastase and tyrosinase activity of Bletillae Rhizoma extract (BR). Methods : The effects of BR on type I procollagen production and collagenase activity in human normal fibroblasts Hs68 after UVB (312 nm) irradiation were measured by ELISA method. The elastase activity, tyrosinase activity, and L-DOPA oxidation after treatment of BR were measured as well. Results : In the present study, the collagen production (type I procollagen) was significantly increased to $15.7{\pm}1.8$ ng/ml at a concentration of BR 100 ${\mu}g/ml$ in UVB damaged Hs68 cells. The increased collagenase activity after UVB damage was significantly recovered to $42.7{\pm}0.7%$, $54.5{\pm}3.5%$, and $38.4{\pm}0.9%$ by BR 10, 30, and 100 ${\mu}g/ml$. The activities of BR 10 mg/ml on tyrosinase activity was significantly reduced to $45.1{\pm}8.4%$ as well. However, there were no significant effects on the elastase activity and the L-DOPA oxidation. Conclusion : BR showed the promoting effects of collagen synthesis and inhibitory effects of collagenase activity in Hs68, human normal fibroblast cells. And these could be thought to have the anti-wrinkle effects and whitening effects in vitro. These results suggest that BR may have potential as an anti-aging ingredient in cosmetic treatment.

Awareness of Reproductive Health Risks, Sex Hormone Levels and Sperm Indices among Farmers Exposed to Pesticides in Akungba Akoko, Nigeria

  • Yeiya, Evelyn Apiriboh;Emokpae, Mathias Abiodun
    • Journal of Environmental Health Sciences
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    • v.48 no.4
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    • pp.244-253
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    • 2022
  • Background: The indiscriminate use of pesticides in Nigeria may have harmful effects on reproductive health of farmers. Objectives: This study assessed the awareness of reproductive health, serum follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone, estradiol, progesterone and sperm characteristics of male farmers occupationally exposed to pesticides. Methods: Eighty four male farmers were recruited for the study. Structured questionnaire was used to obtain the socio-demographic data. Blood and semen samples were collected from the subjects in the morning for hormonal assays and semen analysis using enzyme linked immunosorbent assay (ELISA) method and SQAV sperm quality analyzer. Data were analyzed using chi square, Student's-t-test, and Regression analysis. Results: Serum FSH (p<0.01), LH (p<0.005) and Estradiol (p<0.001) were significantly higher while prolactin (p<0.02) and testosterone (p<0.001) were significantly lower among pesticides exposed farmers than nonexposed subjects. Some 34/84 (40.5%) of the pesticides exposed farmers had serum testosterone levels below the lower limit of the reference range. Those with low testosterone levels (p<0.001), also had FSH (p<0.05), LH (p<0.001) and Estradiol (p<0.002) significantly lower than those with normal testosterone levels. The sperm count among pesticides exposed farmers; total motility and percentage morphology were significantly lower than non-pesticides exposed subjects. Some 14/84 (16.7%) of the pesticides exposed farmers had sperm count below 15 million/mL (oligozoospermia). More than 70% of the farmers were not aware of the reproductive health risks associated with pesticides and only 23.8% of the farmers were using protective devices. Conclusions: Deliberate efforts to improve awareness, knowledge, personal hygiene, and interventions necessary to lessen both pesticides exposure and health risks by adopting safe practices are suggested.