• Title/Summary/Keyword: safety in vivo

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Physical Phenotype Analysis in Obesity Study in vivo (마우스를 이용한 비만연구에서 활용되는 물리적 표현형 분석 기술)

  • Kim, Hyunkyung;Go, Gwang-woong
    • Food Science and Industry
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    • v.51 no.1
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    • pp.37-44
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    • 2018
  • The prevalence of obesity is continually increasing in South Korea; about 1/3 of adults are diagnosed with obesity and 1/2 of adults are overweight in 2016. Abnormal body fat mass increased the risk factors of metabolic syndrome (including hypertension, type II diabetes, dyslipidemia), chronic kidney failure, osteoarthritis, and cardiovascular disease. Ministry of Food and Drug Safety (MFDS) in Korea established the validation and approval system for "functional food" based on related laws and regulations. According to the guideline of MFDS, the biomarkers for obesity study in vitro, in vivo, and clinical trial are well summarized. The analysis of physical phenotypes is necessary condition to study further molecular phenotypes and pathway analysis in vivo study. Thus, we will review currently available physical phenotype analysis; dual energy X-ray absorptiometry (DEXA) and Oxylet gas analysis will be examined in-depth.

Genotoxicological Safety of Gamma-Irradiated Kwamegi(semi-dried Colobabis seira) (감마선 조사된 과메기의 유전독성학적 안전성 평가)

  • 육홍선;정영진;송현파;이주운;변명우
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.1
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    • pp.182-192
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    • 2004
  • Gamma irradiation at 5 and 10 kGy was applied to Kwamegi (semi-dried Colobabis seira) for their possible hygiene quality and carried out genotoxicological safety. In vitro genotoxicological safety of each 5 and 10 kGy-irradiated Kwamegi was evaluated by Salmonella typhimurium (TA98, TA100, TA1535 and TA1537) and E. coli WP2 uvrA reversion assay, SOS chromotest (Escherichia coli PQ37) and chromosome aberration test (Chinese hamster lung fibroblast cells) in the absence and presence of an exogenous metabolizing system (S9 mix). Gamma-irradiated samples were not different from nonirradiated-control to respective in vitro tests. And in vivo micronucleus test using ICR mice (male) micronucleus was not observed. Kwamegi exposed to 10 kGy-gamma ray revealed negative results in these three in vitro mutagenetic tests and in vivo micronucleus test up to 10,000 $\mu\textrm{g}$/plate, respectively. The results indicated that 5 and 10 kGy gamma-irradiated Kwamegi (semi-dried Colobabis seira) did not have mutagenicity.

Viable Alternatives to in vivo Tests for Evaluating the Toxicity of Engineered Carbon Nanotubes

  • Kwon, Soon-Jo;Eo, Soo-Mi
    • Journal of Environmental Health Sciences
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    • v.38 no.1
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    • pp.1-7
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    • 2012
  • Carbon nanotubes (CNTs) stand at the frontier of nanotechnology and are destined to stimulate the next industrial revolution. Rapid increase in their production and use in the technology industry have led to concerns over the effects of CNT on human health and the environment. The prominent use of CNTs in biomedical applications also increases the possibility of human exposure, while properties such as their high aspect ratio (fiber-like shape) and large surface area raise safety concerns for human health if exposure does occur. It is crucial to develop viable alternatives to in vivo tests in order to evaluate the toxicity of engineered CNTs and develop validated experimental models capable of identifying CNTs' toxic effects and predicting their level of toxicity in the human respiratory system. Human lung epithelial cells serve as a barrier at the interface between the surrounding air and lung tissues in response to exogenous particles such as air-pollutants, including CNTs. Monolayer culture of the key individual cell types has provided abundant fundamental information on the response of these cells to external perturbations. However, such systems are limited by the absence of cell-cell interactions and their dynamic nature, which are both present in vivo. In this review, we suggested two viable alternatives to in vivo tests to evaluate the health risk of human exposure to CNTs.

In vitro and in vivo application of anti-cotinine antibody and cotinine-conjugated compounds

  • Kim, Hyori;Yoon, Soomin;Chung, Junho
    • BMB Reports
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    • v.47 no.3
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    • pp.130-134
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    • 2014
  • The combination of a high-affinity antibody to a hapten, and hapten-conjugated compounds, can provide an alternative to the direct chemical cross-linking of the antibody and compounds. An optimal hapten for in vitro use is one that is absent in biological systems. For in vivo applications, additional characteristics such as pharmacological safety and physiological inertness would be beneficial. Additionally, methods for cross-linking the hapten to various chemical compounds should be available. Cotinine, a major metabolite of nicotine, is considered advantageous in these aspects. A high-affinity anti-cotinine recombinant antibody has recently become available, and can be converted into various formats, including a bispecific antibody. The bispecific anti-cotinine antibody was successfully applied to immunoblot, enzyme immunoassay, immunoaffinity purification, and pre-targeted in vivo radioimmunoimaging. The anti-cotinine IgG molecule could be complexed with aptamers to form a novel affinity unit, and extended the in vivo half-life of aptamers, opening up the possibility of applying the same strategy to therapeutic peptides and chemical compounds.

Evaluation of the anti-Toxoplasma gondii Activity of Hederagenin in vitro and in vivo

  • Zhang, Run-Hui;Jin, Runhao;Deng, Hao;Shen, Qing-Kun;Quan, Zhe-Shan;Jin, Chun-Mei
    • Parasites, Hosts and Diseases
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    • v.59 no.3
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    • pp.297-301
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    • 2021
  • Toxoplasma gondii infection is widespread worldwide, not only posing a serious threat to human food safety and animal husbandry, but also endangering human health. The selectivity index was employed to measure anti-T. gondii activity. Hederagenin (HE) exhibited potent anti-T. gondii activity and low cytotoxicity. For this reason, HE was selected for in vivo experiments. HE showed 64.8%±13.1% inhibition for peritoneal tachyzoites in mice, higher than spiramycin 56.8%±6.0%. Biochemical parameters such as alanine aminotransferase, aspartate aminotransferase, glutathione, and malondialdehyde, illustrated that HE was a good inhibitor of T. gondii in vivo. This compound was also effective in relieving T. gondii-induced liver damage. Collectively, it was demonstrated that HE had potential as an anti-T. gondii agent.

Ruminal pH pattern, fermentation characteristics and related bacteria in response to dietary live yeast (Saccharomyces cerevisiae) supplementation in beef cattle

  • Zhang, Xiangfei;Dong, Xianwen;Wanapat, Metha;Shah, Ali Mujtaba;Luo, Xiaolin;Peng, Quanhui;Kang, Kun;Hu, Rui;Guan, Jiuqiang;Wang, Zhisheng
    • Animal Bioscience
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    • v.35 no.2
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    • pp.184-195
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    • 2022
  • Objective: In this study we aimed to evaluate the effect of dietary live yeast supplementation on ruminal pH pattern, fermentation characteristics and associated bacteria in beef cattle. Methods: This work comprised of in vitro and in vivo experiments. In vitro fermentation was conducted by incubating 0%, 0.05%, 0.075%, 0.1%, 0.125%, and 0.15% active dried yeast (Saccharomyces cerevisiae, ADY) with total mixed ration substrate to determine its dose effect. According to in vitro results, 0.1% ADY inclusion level was assigned in in vivo study for continuously monitoring ruminal fermentation characteristics and microbes. Six ruminally cannulated steers were randomly assigned to 2 treatments (Control and ADY supplementation) as two-period crossover design (30-day). Blood samples were harvested before-feeding and rumen fluid was sampled at 0, 3, 6, 9, and 12 h post-feeding on 30 d. Results: After 24 h in vitro fermentation, pH and gas production were increased at 0.1% ADY where ammonia nitrogen and microbial crude protein also displayed lowest and peak values, respectively. Acetate, butyrate and total volatile fatty acids concentrations heightened with increasing ADY doses and plateaued at high levels, while acetate to propionate ratio was decreased accordingly. In in vivo study, ruminal pH was increased with ADY supplementation that also elevated acetate and propionate. Conversely, ADY reduced lactate level by dampening Streptococcus bovis and inducing greater Selenomonas ruminantium and Megasphaera elsdenii populations involved in lactate utilization. The serum urea nitrogen decreased, whereas glucose, albumin and total protein concentrations were increased with ADY supplementation. Conclusion: The results demonstrated dietary ADY improved ruminal fermentation dose-dependently. The ruminal lactate reduction through modification of lactate metabolic bacteria could be an important reason for rumen pH stabilization induced by ADY. ADY supplementation offered a complementary probiotics strategy in improving gluconeogenesis and nitrogen metabolism of beef cattle, potentially resulted from optimized rumen pH and fermentation.

Potential Probiotic Characteristics and Safety Assessment of Lactobacillus rhamnosus SKG34 Isolated from Sumbawa Mare's Milk

  • Sujaya, I Nengah;Suwardana, Gede Ngurah Rsi;Gotoh, Kazuyoshi;Sumardika, I Wayan;Nocianitri, Komang Ayu;Sriwidyani, Ni Putu;Putra, I Wayan Gede Artawan Eka;Sakaguchi, Masakiyo;Fatmawati, Ni Nengah Dwi
    • Microbiology and Biotechnology Letters
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    • v.50 no.1
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    • pp.51-62
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    • 2022
  • Lactobacillus rhamnosus SKG34 (LrSKG34), a potential probiotic strain, was successfully isolated from Sumbawa Mare's milk. Our previous studies showed that the strain is resistant to gastrointestinal conditions, possesses antioxidant activity, and lowers blood cholesterol levels. Further clarification of the potential probiotic characteristics and safety assessment are necessary. This study aimed to evaluate the adhesion of LrSKG34 to Caco-2 cell monolayers and its effect on mucosal integrity in vitro. We also examined the LrSKG34 safety profile based on antimicrobial susceptibility testing, haemolytic activity determination, Caco-2 cell monolayer translocation evaluation, and in vivo investigation of the effect of LrSKG34 on the physiology, biochemical markers, and histopathological appearance of major organs in an animal model. LrSKG34 attached to Caco-2 cell monolayers and maintained mucosal integrity in vitro. The typical resistance of lactobacilli to ciprofloxacin, gentamicin, vancomycin, trimethoprim-sulfamethoxazole, and metronidazole was confirmed for LrSKG34. No haemolytic activity was observed on blood agar plates, and no LrSKG34 translocation was observed in Caco-2 cell monolayers. Administration of LrSKG34 to Sprague-Dawley rats did not adversely affect body weight. No abnormalities in hematological parameters, serum biochemistry levels, or histopathological structures of major organs were observed in LrSKG34-treated rats. Collectively, the results implicate LrSKG34 as a promising and potentially safe probiotic candidate for further development.

Development of a Test Method for the Evaluation of DNA Damage in Mouse Spermatogonial Stem Cells

  • Jeon, Hye Lyun;Yi, Jung-Sun;Kim, Tae Sung;Oh, Youkyung;Lee, Hye Jeong;Lee, Minseong;Bang, Jin Seok;Ko, Kinarm;Ahn, Il Young;Ko, Kyungyuk;Kim, Joohwan;Park, Hye-Kyung;Lee, Jong Kwon;Sohn, Soo Jung
    • Toxicological Research
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    • v.33 no.2
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    • pp.107-118
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    • 2017
  • Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration ($IC_{50}$) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.

Genotoxicological Safety of the Gamma-Irradiated Medicinal Herbs (감마선 조사 생약재의 안전성에 관한 유전독성학적 평가)

  • 조성기
    • Journal of Food Hygiene and Safety
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    • v.12 no.3
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    • pp.217-227
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    • 1997
  • These experiments were performed to investigate the safety of the three medicinal herbs- Curcuma longa Linne, Paeonia japonica Miyabe, Scutellaria baikalensis George-irradiated with gamma rays in respect of genotoxicity. The methanol-soluble and water-soluble fractions of the methanol-water extracts of the 10 kGy gamma-irradiated herbs were examined in two short-term in vitro tests : (1) Salmonella typhimurium reversion assay (Ames test) in strain TA 98, TA 100 and TA 012 (2) Micronucleus test in cultured Chinese hamster ovary (CHO) cells. No mutagenicity was detected in the assays with or without metabolic activation. From these results, the safety of the herbs irradiated with gamma rays at practical doses could be revealed in further tests of genotoxicity in vivo, chronic and reproductive toxiceity.

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