• 제목/요약/키워드: Urea assay

검색결과 45건 처리시간 0.032초

Evaluation of Invasive and Noninvasive Methods for the Diagnosis of Helicobacter Pylori Infection

  • Cosgun, Yasemin;Yildirim, Abdullah;Yucel, Mihriban;Karakoc, Ayse Esra;Koca, Gokhan;Gonultas, Alpaslan;Gursoy, Gul;Ustun, Huseyin;Korkmaz, Meliha
    • Asian Pacific Journal of Cancer Prevention
    • /
    • 제17권12호
    • /
    • pp.5265-5272
    • /
    • 2016
  • Objective: The present study was conducted to evaluate invasive and noninvasive diagnostic methods for detection of Helicobacter pylori (H. pylori) in patients admitted with dyspeptic complaints and to compare sensitivities and specificities. Method: Sets of four gastric biopsy specimens were obtained from a total of 126 patients included in the study. The presence of H. pylori was determined by invasive tests including culture, rapid urease test, polymerase chain reaction (PCR) and histopathology. Among noninvasive tests, urea breath test, serological tests and enzyme-linked immunosorbent assay (ELISA) were performed. Results: H. pylori was isolated in 79 (62.7%) gastric biopsy cultures, whereas positivity was concluded for 105 (83.3%) patients by rapid urease test, for 106 (84.1%) by PCR, for 110 (87.3%) by histopathology, for 119 (94.4%) by urea breath test, and for 107 (84.9%) by ELISA. In the present study, the culture findings and histopathological examination findings were accepted as gold standard. According to the gold standard, urea breath test had the highest sensitivity (96.5%) and the lowest specificity (30%), whereas culture and histopathology had the highest specificities (100%). Conclusion: The use of PCR invasively with gastric biopsy samples yielded parallel results with the gold standard. PCR can be recommended for routine use in the diagnosis of H. pylori.

식품 중 대두단백질의 정량분석을 위한 효소면역측정법 (An Enzyme-Linked Immunosorbent Assay for Quantitation of Soy Proteins in Food)

  • 손동화;김현정;음병욱;김수호;김순미
    • 한국식품과학회지
    • /
    • 제32권5호
    • /
    • pp.991-996
    • /
    • 2000
  • 식품 중 대두단백질의 분석을 위한 효소면역측정법(ELISA)을 개발하고자 하였다. 대두단백질의 주 구성 성분이며 열에 안정한 glycinin의 acidic subunits(AS)에 대한 특이항체를 생산하였고 이를 이용하여 간접경합 ELISA(ciELISA)를 확립하였다. 시료처리조건은 urea와 DTT로 처리함으로써 용해시킨 다음 $100^{\circ}C$에서 1시간 열처리하였고 cystine을 함유한 용액으로 재생시켰다. 이러한 시료의 처리조건하에서 ISP를 $60-90^{\circ}C$까지 열처리한 시료에 대해서도 분석결과의 변화가 거의 없는 것으로 나타났다. 표준곡선 상에서 ISP의 검출한계는 0.3 ${\mu}g/mL$이었다. 항AS 항체의 다른 단백질에 대한 반응성을 검토한 결과, 탈지분유, 카제인, 난백분말, ovalbumin에서 0.1%이하로 미약하게 반응하거나 거의 반응하지 않는 것으로 나타났다. ISP를 0.5-2%첨가한 탈지분유에 대한 ISP 분석 회수율은 평균 76%(C.V., 11.5%)였으며 ISP를 0.5-3% 첨가하여 시험 제조한 소세지의 경우 평균 83%(C.V., 19%)로 나타났고 시판 소세지 6점에 함유된 ISP의 함량은 평균 1.27%로 나타났다.

  • PDF

Anti-Helicobacter and Anti-inflammatory Effects of Sohamhyungtang in Helicobacter pylori-Infected Human Gastric Epithelial AGS cells

  • Won, SangBum;Yim, Dongsool;Choi, SungSook
    • Natural Product Sciences
    • /
    • 제23권3호
    • /
    • pp.175-182
    • /
    • 2017
  • This study evaluated the anti-Helicobacter and anti-inflammatory effects of Sohamhyungtang (SHHT). The minimum inhibitory concentration (MIC) of SHHT against Helicobacter pylori (H. pylori) was determined by the agar dilution method. Expression of the H. pylori cagA gene in the presence of SHHT was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Inhibition of H. pylori urease by SHHT was determined by the phenol-hypochlorite assay. Antiadhesion activity of SHHT was measured by urea-phenol red reagent. Inhibition of nitric oxide (NO) production in AGS cells was measured with Griess reagent. Inducible nitric oxide synthase (iNOS) and IL-8 mRNA expression in AGS cells which were infected with H. pylori was determined by qRT-PCR. IL-8 level was measured by enzyme-linked immunosorbent assay (ELISA). The MIC of SHHT was $100{\mu}g/mL$ and the expression of cagA gene was decreased about 25 folds in the presence of SHHT. H. pylori urease was inhibited 90% by SHHT. SHHT inhibited H. pylori adhesion on AGS cell in a concentration dependent manner. mRNA expression of iNOS and IL-8 and the production of NO and IL-8 were significantly decreased in the presence of SHHT. In conclusion, SHHT showed anti-Helicobacter activity and has potent anti-inflammatory effect on H. pylori-induced inflammation in human gastric epithelial AGS cells.

Diurnal Variations in Milk and Blood Urea Nitrogen and Whole Blood Ammonia Nitrogen in Dairy Cows

  • Hwang, Sen-Yuan;Lee, Mei-Ju;Peh, Huo-Cheng
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제14권12호
    • /
    • pp.1683-1689
    • /
    • 2001
  • The levels of urea nitrogen both in blood (BUN) and milk (MUN), and milk protein (MP) reflect protein and energy intake in dairy herd feeding. Blood and milk constituents may be changes rhythmically and influence by different sampling time within a day and after feeding. Trials were conducted using five dietary treatments in both lactating and dry cows to study the effects of sampling time on concentrations of BUN, MUN and whole blood ammonia nitrogen (BAN) in practical dairy cow feeding in Taiwan. The conventional feed ingredients and forages including corn silage, alfalfa hay, timothy or pangola hay and corn grain were used as major source of the diet to follow practical dairy cow feeding. Five different diets were varying in amounts (low=L; standard=S; high=H) of crude protein (P) and energy (E) according to the NRC (1989). The energy to protein ratios in kcal/kg for the PSES, PLES, PHES, PSEH and PSEL were 10.82, 12.54, 9.41, 12.53 and 9.13 in lactating cows, and 11.38, 13.33, 9.78, 13.28 and 9.74 in dry cows, respectively. Results showed that after feeding at 9:30, BUN reached peak at 13:30 and was significantly higher than those to that sampled at 14:30 to 18:30 (p<0.05) in dry cows. Therefore the best blood sampling time for urea nitrogen assay in dry cows is 4 hours after morning feeding. In lactating cows, BUN of 13:30 was significantly higher than those of 8:30 to 11:30 (p<0.05), but there were no significant difference between the BUN values of other sampling time. Hence the suitable blood sampling time for BUN value in lactating cows was located on 3 to 8 hours after morning feeding, but the best time was 4 hours after morning feeding. MUN content is significantly higher in the afternoon collected bulk milk than the fore-strip morning milk (p<0.05), therefore the best sampling time for MUN is from afternoon collected bulk milk. Diurnal BAN changed without traceable rhythmic pattern and was negatively correlated to the BUN (r = -0.78). It is suggested that BAN may not be a good indicator for monitoring dairy cow feeding.

Evaluation of the Genetic Toxicity of Synthetic Chemicals (XI) - a Synthetic Sulfonylurea Herbicide, Pyrazosulfuron-ethyl-

  • Ryu, Jae-Chun;Kim, Eun-Young;Kim, Young-Seok;Yun, Hye-Jung
    • 한국환경성돌연변이발암원학회지
    • /
    • 제24권1호
    • /
    • pp.33-39
    • /
    • 2004
  • To validate and to estimate the chemical hazard playa very important role to environment and human health. The detection of many synthetic chemicals including agrochemicals that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. Pyrazosulfuron-ethyl [Ethyl-5-(4,6-dimethoxypyrimidin-2-ylcarbamoylsulfamoyl)-1-methylpyrazole-4-carboxylate, $C_{14}H_{18}N{6}O_{7}S,$ M.W. =414.39, CAS No. 93697-74-6], is one of well known rice herbicide belong in the sulfonyl urea group. To clarify the genotoxicity of this agrochemical, Ames bacterial reversion assay, in vitro chromosomal aberration assay with Chinese hamster lung (CHL) fibroblast and bone marrow micronucleus assay in mice were subjected. In Ames assay, although pyrazosulfuron-ethyl revealed cytotoxic at 5,000-140 $\mug/plate$ in Salmonella typhimurium TA100, no dose-dependent mutagenic potential in 4.4~70 $\mug/plate$ of S. typhimurium TA 98, TA 100, TA1535 and TA 1537 both in the absence and presence of S-9 metabolic activation system was observed. Using CHL fibroblasts, the 50% cell growth inhibition concentration $(IC_{50})$ of pyrazosulfuron-ethyl was determined as 1,243 $\mug/mL,$ and no chromosomal aberration was observed both in the absence and presence of S-9 mixture in the concentration range of 311-1,243 $\mug/mL.$ And also, in vivo micronucleus assay using mouse bone marrow, pyrazosulfuron-ethyl revealed no remarkable induction of MNPCE (micronucleated polychromatic erythrocytes/1000 polychromatic erythrocytes) in the dose range of 625-2,500 mg/kg body weight when administered orally. Consequently, Ames bacterial gene mutation with Salmonella typhimurium, in vitro chromosome aberration with mammalian cells and in vivo bone marrow micronucleus assay revealed no clastogenic potential of pyrazosulfuron-ethyl in this study.

  • PDF

돼지에서 신장 허혈 관류 손상에 미치는 환원형 L-glutathione의 효과 (The Effects of Reduced L-glutathione on Renal Ischemia-Reperfusion Injury in Pigs)

  • 이재연;김현수;지현철;정성목;조성환;박창식;김명철
    • 한국임상수의학회지
    • /
    • 제26권3호
    • /
    • pp.200-204
    • /
    • 2009
  • This study was performed to evaluate the effects of reduced L-glutathione on the oxidant/antioxidant status(superoxide dismutase(SOD), catalase(CAT), glutathione peroxidase(GPx), protein carbonyl and lipid hydroperoxide(LPO) concentration), renal function(blood urea nitrogen(BUN) and serum creatinine levels), and microscopy of renal tissues in pigs undergoing unilateral renal ischemia-reperfusion(I/R). Sixteen Landrace and Yorkshire mixed-breed pigs were divided randomly into two groups: untreated control group and reduced L-glutathione-treated group(4 mg/kg IV). Each group had 8 pigs. Pigs were unilaterally nephrectomized and the kidney was subject to 30 min of renal pedicle occlusion. Blood samples for biochemical assay were collected on days 1, 3, 5, 7, and 14 post nephrectomy. Renal I/R injury were evaluated histopathologically by the microscopic observation of renal tissue sections and biochemically by the measurement of the plasma creatinine and urea levels. Parameters of oxidative stress such as SOD, GPx, CAT, protein carbonyl and LPO were measured. The elevation of creatine and BUN levels was lower in the treated group, compared with the control group. The activities of antioxidant-enzyme were higher in the treated group, compared with the control group. In histological findings, the severity of damage in the reduced L-glutathione treated group was less when compared to the control group.

Protective effect of ginsenosides Rk3 and Rh4 on cisplatin-induced acute kidney injury in vitro and in vivo

  • Baek, Seung-Hoon;Shin, Byong-kyu;Kim, Nam Jae;Chang, Sun-Young;Park, Jeong Hill
    • Journal of Ginseng Research
    • /
    • 제41권3호
    • /
    • pp.233-239
    • /
    • 2017
  • Background: Nephrotoxicity is the major side effect in cisplatin chemotherapy. Previously, we reported that the ginsenosides Rk3 and Rh4 reduced cisplatin toxicity on porcine renal proximal epithelial tubular cells (LLC-PK1). Here, we aimed to evaluate the protective effect of ginsenosides Rk3 and Rh4 on kidney function and elucidate their antioxidant effect using in vitro and in vivo models of cisplatin-induced acute renal failure. Methods: An enriched mixture of ginsenosides Rk3 and Rh4 (KG-KH; 49.3% and 43.1%, respectively) was purified from sun ginseng (heat processed Panax ginseng). Cytotoxicity was induced by treatment of $20{\mu}M$ cisplatin to LLC-PK1 cells and rat model of acute renal failure was generated by single intraperitoneal injection of 5 mg/kg cisplatin. Protective effects were assessed by determining cell viability, reactive oxygen species generation, blood urea nitrogen, serum creatinine, antioxidant enzyme activity, and histopathological examination. Results: The in vitro assay demonstrated that KG-KH ($50{\mu}g/mL$) significantly increased cell viability (4.6-fold), superoxide dismutase activity (2.8-fold), and glutathione reductase activity (1.5-fold), but reduced reactive oxygen species generation (56%) compared to cisplatin control cells. KG-KH (6 mg/kg, per os) also significantly inhibited renal edema (87% kidney index) and dysfunction (71.4% blood urea nitrogen, 67.4% creatinine) compared to cisplatin control rats. Of note, KG-KH significantly recovered the kidney levels of catalase (1.2-fold) and superoxide dismutase (1.5-fold). Conclusion: Considering the oxidative injury as an early trigger of cisplatin nephrotoxicity, our findings suggest that ginsenosides Rk3 and Rh4 protect the kidney from cisplatin-induced oxidative injury and help to recover renal function by restoring intrinsic antioxidant defenses.

Melittin induces autophagy to alleviate chronic renal failure in 5/6-nephrectomized rats and angiotensin II-induced damage in podocytes

  • Yufan Zhang;Huaping Xu;Hongwei Qiao;Ya Zhao;Minmin Jiang
    • Nutrition Research and Practice
    • /
    • 제18권2호
    • /
    • pp.210-222
    • /
    • 2024
  • BACKGROUND/OBJECTIVES: Chronic renal failure (CRF) is a complex pathological condition that lacks a cure. Certain Chinese medicines, such as melittin, a major component in bee venom, have shown efficacy in treating CRF patients. On the other hand, the mechanisms underlying the therapeutic effects of melittin are unclear. MATERIALS/METHODS: A 5/6 nephrectomy model (5/6 Nx) of renal failure was established on rats for in vivo assays, and mouse podocyte clone 5 (MPC5) mouse podocyte cells were treated with angiotensin II (AngII) to establish an in vitro podocyte damage model. The 24-h urine protein, serum creatinine, and blood urea nitrogen levels were evaluated after one, 2, and 4 weeks. Hematoxylin and eosin staining, Masson staining, and periodic acid-Schiff staining were used to examine the pathological changes in kidney tissues. A cell counting kit 8 assay was used to assess the cell viability. Reverse transcription polymerase chain reaction and Western blot were used to assess the mRNA and protein levels in the cells, respectively. RESULTS: In the rat 5/6 Nx, melittin reduced the 24-h urinary protein excretion and the serum creatinine and blood urea nitrogen levels. Furthermore, the renal pathology was improved in the melittin-treated 5/6 Nx rats. Melittin promoted podocin, nephrin, Beclin 1, and the LC3II/LC3I ratio and inhibited phosphorylated mammalian target of rapamycin (mTOR)/mTOR in 5/6 Nx-induced rats and AngII-induced MPC5 mouse podocyte cells. Moreover, inhibiting autophagy with 3-MA weakened the effects of melittin on podocin, nephrin, and the LC3II/LC3I ratio in podocytes. CONCLUSION: Melittin may offer protection against kidney injury, probably by regulating podocyte autophagy. These results provide the theoretical basis for applying melittin in CRF therapy.

새로운 CPS1 유전자 돌연변이에 의한 신생아형 carbamoyl phosphate synthetase 1 결핍 1례 (A Case of Lethal Neonatal Type Carbamoyl Phosphate Synthetase 1 Deficiency with Novel Mutation of CPS1)

  • 서승현;김유미;변신연;손승국;김성헌;김형태;김구환;유한욱
    • 대한유전성대사질환학회지
    • /
    • 제16권2호
    • /
    • pp.109-114
    • /
    • 2016
  • Carbamoyl phosphate synthetase 1 (CPS1) 결핍은 상염색체 열성 유전을 하는 매우 드문 유전질환으로, 요소 회로의 첫 번째 효소인 carbamoyl phosphate synthetase의 결핍에 의해 고암모니아혈증을 유발한다. CPS1 결핍은 신생아시기부터 성인까지 다양한 시기에 고암모니아혈증이 발현될 수 있으나 대부분 신생아 시기의 치명적인 고암모니아혈증으로 발현하여 예후가 불량하며 응급 투석 및 집중 치료가 필요하다. 본 증례는 드문 유전 질환인 CPS1 결핍 신생아에서 새로운 CPS1 유전자의 돌연변이를 발견하였고 조직적인 팀 접근을 통해 심각한 고암모니아혈증에 대한 신속한 투석 및 집중 치료를 시행하여 심한 뇌 병변 및 사망을 예방하고 양호한 경과를 보였음을 보고하는 바이다.

  • PDF

Investigation of Goats' Milk Adulteration with Cows' Milk by PCR

  • Cheng, Yeong-Hsiang;Chen, Su-Der;Weng, Ching-Feng
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제19권10호
    • /
    • pp.1503-1507
    • /
    • 2006
  • Goats' milk adulteration with cows' milk is becoming a big problem. In the past, the urea-polyacrylamide gel electrophoresis assay with different motility of ${\alpha}S1$-casein has been applied for the identification of cows' milk adulteration. The detection sensitivity is 1.0%. The aim of this study was to develop a faster and more sensitive method to detect cows' milk which may be present in adulterated goats' milk and goats' milk powder. The published primer was targeted at highly conserved regions in bovine mitochondrial DNA (a 271 bp amplicon). This amplicon was cloned and sequenced to further confirm bovine specific sequence. The chelex-100 was used to separate bovine somatic cells from goats' milk or goats' milk powder samples. Random sampling of different brands of goats' milk powder and tablets from various regions of Taiwan showed the adulterated rate was 20 out of 80 (25%) in goats' milk powders and 12 out of 24 (50%) in goats' milk tablets. With this system, as low as 0.1% cows' milk or cows' milk powder in goat milk or goat milk powder could be identified. This chelex DNA isolation approach provides a fast, highly reproducible and sensitive method for detecting the adulteration of goats' milk products.