Cadmium is one of the well-known environmental toxicants and induces cancer in rodents and human, but its carcinogenic mechanism has not been well demonstrated until now. Genotoxic effects of cadmium in Salmonella typhimurium TA98, TA100 and TA1535/pSK1002 or in WB-F344 rat liver epithelial cells were investigated to elucidate the tumor initiating effects of cadmium. TA98, TA100 and TA1535/pSK1002 tester strains were used to detect frameshift mutation, base-pair mutation and SOS repair response, respectively, in Salmonella mutation test. Reverse mutations from histidine to $histidin^+$ of Salmonella typhimurium TA98 and TA100 by $CdCl_2$ were not significantly different from control up to the maximum doses ($100{\mu}M$ and $200{\mu}M$ in TA98 and TA100, respectively) at which non-cytotoxicity was observed. DNA SOS repair responses(${\beta}$-galactosidase activity) generally did not show significant increases compared to control in both of the conditions with or without metabolic activation in Salmonella typhimurium TA1535/pSK1002 by $CdCl_2$. But the activities of ${\beta}$-galactosidase by $400{\mu}M$ of $CdCl_2$ in metabolic activation condition and by 130 and $400{\mu}M$ of $CdCl_2$ in non-metabolic activation condition were more decreased than those of control. DNA single strand breaks for 4hrs were observed only in WB-F344 rat liver epithelial cells treated with $200{\mu}M$ of $CdCl_2$. As a conclusion, $CdCl_2$ did not induce gene mutation in microbials but induce DNA single strand breaks in rat liver epithelial cells.
The aim of this study was 1) to confirm the practical efficiency of a routine milk P4 monitoring system for postpartum reproductive management of a dairy herd, and 2) to evaluate the relationship between the blood metabolic profiles, milk quality and body weight of individual cows in the farm records, which may reflect the postpartum nutritional condition, and the time of postpartum resumption of ovarian activity of dairy cows. A total of 116 Holstein cows was used in the present study. First, during the period of Experiment 1, postpartum reproductive management based on weekly measured milk P4 concentration from individual cows was conducted. Compared with the reproductive records of the past two years without P4 monitoring, although the day from calving to first AI did not change, both the number of AI until pregnant (with P4; 1.9 times vs. without P4; 2.9 times) and the days open (with P4; 95.1 days vs. without P4; 135.8 days and 133.8 days) were significantly decreased. In Experiment 2, the measurement of blood constituents such as albumin, blood urea nitrogen, packed cell volume, ammonia, glucose, total cholesterol, non-esterified, AST and $\gamma$-GTP was performed on the blood samples taken once approximately 14 days postpartum, to monitor both health and nutritional conditions. The milk constituent parameters, such as milk protein (MP), milk fat (MF), SNF and lactose, collected from the monthly progeny test of individual cows, were used to monitor the postpartum nutritional status. Furthermore, the data obtained from the routine measurements of body weight were used to calculate the rate of peripartum body weight loss. The resumption day of the postpartum estrous cycle was assumed from the milk P4 profiles of individual cows. There was no clear relationship between each parameter from blood examination and those from resumption time. However, the cows had low values of MP, and SNF, which significantly affected the resumption of the postpartum estrous cycle. Similarly, a higher rate of body weight loss indicated a significant delay (more than 1 month) in the resumption of the postpartum estrous cycle, compared with the groups that had a medium or lower rate of body weight loss. The results of the present study demonstrated that the implementation of routine milk P4 monitoring-based postpartum reproductive management, together with milk quality parameters and routine BW data available in field conditions may be utilized as a practical approach for increasing the postpartum reproductive efficiency of a high yielding dairy herd.
Raman microspectroscopy is a promising tool for microbial analysis at single cell level since it can rapidly measure the cell materials including lipids, nucleic acids, and proteins by measuring the inelastic scattering of a molecule irradiated by monochromatic lights. Using Raman spectra provides high specificity and sensitivity in classification of bacteria at the strain level. In addition, a Raman approach coupled with stabled isotope such as $^{13}C$ and $^2H$ is able to detect and quantify general metabolic activity at single cell level. After bacterial detection process by Raman microspectroscopy, interested unculturable cell sorting and single cell genomics can be accomplished by combination with optical tweezer and microfluidic devices. In this review, the characteristics and applications of Raman microspectroscopy were reviewed and summarized in order to provide a better understanding of microbial analysis using Raman spectroscopy.
Objective: This study aimed to determine the protective efficacy of Buddha's Temple (BT) extract against tert-butyl hydroperoxide (t-BHP)-induced oxidative stress in Gallus gallus chicken embryo fibroblast cell line (DF-1) and its effects on the cell lipid metabolism. Methods: In this experimental study, Gallus gallus DF-1 fibroblast cells were pretreated with BT 10-7 for 24 hours, followed by their six-hour exposure to t-BHP (100 μM). Water-soluble tetrazolium salt-8 (WST-8) assays were performed, and the growth curve was computed. The intracellular gene expression changes caused by BT extract were confirmed through quantitative polymerase chain reaction (qPCR). Flow cytometry, oil red O staining experiment, and thin-layer chromatography were performed for the detection of intracellular metabolic mechanism changes. Results: The WST-8 assay results showed that the BT pretreatment of Gallus gallus DF-1 fibroblast cell increased their cell survival rate by 1.08%±0.04%, decreased the reactive oxygen species (ROS) level by 0.93%±0.12% even after exposure to oxidants, and stabilized mitochondrial activity by 1.37%±0.36%. In addition, qPCR results confirmed that the gene expression levels of tumor necrosis factor α (TNFα), TIR domain-containing adapter inducing IFN-beta (TICAM1), and glucose-regulated protein 78 (GRP78) were regulated, which contributed to cell stabilization. Thin-layer chromatography and oil red O analyses showed a clear decrease in the contents of lipid metabolites such as triacylglycerol and free fatty acids. Conclusion: In this study, we confirmed that the examined BT extract exerted selective protective effects on Gallus gallus DF-1 fibroblast cells against cell damage caused by t-BHP, which is a strong oxidative inducer. Furthermore, we established that this extract significantly reduced the intracellular ROS accumulation due to oxidative stress, which contributes to an increase in poultry production and higher incomes.
Journal of Physiology & Pathology in Korean Medicine
/
v.21
no.6
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pp.1506-1512
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2007
In order to investigate the totale yield, the content of soluble polysaccharide and the others of FEW extract from the yeast fermentated Ganoderma lucidium by supersonic method, the yeast strain was inoculated after pretreatment and subsequently followed fermentation and supersonic extract. The main construction of the extract method is composed of the main glucose and together with the xylose, fucose, galactose and mannose. Results show that because of the generated lactic acid and ethanol, pH value of extract decreases and the safety as well as the preservation is improved. The extract yield, the total soluble polysaccharide, SOD-like activity, cytotoxic effect and growth inhibitory effect against cancer cell line are much higher in FE method than RE method, especially FEW3 extracts fermented during 24hrs. It is concluded that yeast fermentation makes the extract yield increase because of the cell disintegration, the useful ingredients of the germ body, the metabolic products, the insoluble ingredients due to the generation of ethanol, and the cell fragmentation caused by the supersonic waves vibration. Content of generated ethanol, total soluble polysaccharide and extract yield all increase during the fermentation time from 24 to 72 hours and the optimum fermentation condition is at $27^{\circ}C$ for 72 hours. The bitter taste and smell of the Ganoderma lucidium extract is diminished, fragrant-bitter taste and smell is generated so that the whole functional quality is improved.
N-acetylsphingosine (C2-ceramide) is a synthetic water-soluble ceramide mimicking the activity of natural ceramides. By fixing chiral conformation on carbon numbers 2 and 3 in the ceramide structure, four chiral C2-ceramides naming d-erythro-, I-erythro-, d-threo-and 1-three C2-ceramide were synthesized. We have investigated the chiral effects of these C2-ceramides on the sphingolipid metabolism, particularly on both the sphingolipid bio- synthetic pathway and on the degradation pathway. In both HL-60 and U937 cells, the chiral C2-ceramide ($10{\mu}\textrm{m}$) showed sphingosine accumulation monitored fluoromatrically by a high performance liquid chromatographic separation of the sphingoid bases. Most importantly, in HL-60 cells, l-erythro C2-ceramide induced a 50 fold increase in sphingosine as compared to the control, while l-threo C2-ceramide exhibited a minimal 7-fold in-crease. In contrast, sphinganine, another sphingoid base, showed less accumulation by any chiral C2-ceramide tested under the same conditions. These results suggested that chiral C2-ceramide primarilyacts on the sphingolipid degradation pathway rather than on the sphingolipid biosynthetic route. The strong $C_0/G_1$ phase arrest in the cell cycle by treatment of I-erythro C2-ceramide indicates that the blockade of the sphingolipid degradation pathway might be concomitantly involved in the dysfunction of the cell cycle. On the other hand, the fact that all chiral C2-ceramides tested failed to inhibit the activity of sphingosine kinase acting on the removal of sphingosine by producing sphingosine-1 -phosphate demonstrates that chiral C2- ceramides may increase sphingosine by activating various ceramidases by which natural ceramides are divided into sphingosine and free fatty acids. However, the precise steps involved in this interaction are still unknown.
Background: Rb3 is a ginsenoside with anti-inflammatory properties in many cell types and has been reported to attenuate inflammation-related metabolic diseases such as insulin resistance, nonalcoholic fatty liver disease, and cardiovascular disease. However, the effect of Rb3 on podocyte apoptosis under hyperlipidemic conditions, which contributes to the development of obesity-mediated renal disease, remains unclear. In the current study, we aimed to investigate the effect of Rb3 on podocyte apoptosis in the presence of palmitate and explore its underlying molecular mechanisms. Methods: Human podocytes (CIHP-1 cells) were exposed to Rb3 in the presence of palmitate as a model of hyperlipidemia. Cell viability was assessed by MTT assay. The effects of Rb3 on the expression of various proteins were analyzed by Western blotting. Apoptosis levels were determined by MTT assay, caspase 3 activity assay, and cleaved caspase 3 expression. Results: We found that Rb3 treatment alleviated the impairment of cell viability and increased caspase 3 activity as well as inflammatory markers in palmitate-treated podocytes. Treatment with Rb3 dosedependently increased PPARδ and SIRT6 expression. Knockdown of PPARδ or SIRT6 reduced the effects of Rb3 on apoptosis as well as inflammation and oxidative stress in cultured podocytes. Conclusions: The current results suggest that Rb3 alleviates inflammation and oxidative stress via PPARδ-or SIRT6-mediated signaling, thereby attenuating apoptosis in podocytes in the presence of palmitate. The present study provides Rb3 as an effective strategy for treating obesity-mediated renal injury.
Ha, Eun-Suk;Hwang, Soo-Hyun;Shin, Kwang-Soon;Yu, Kwang-Won;Lee, Keyong-Ho;Choi, Joo-Sun;Park, Woo-Mun;Yoon, Taek-Joon
Archives of Pharmacal Research
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v.27
no.2
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pp.217-224
/
2004
Previously, we reported that water-extracted Acanthopanax senticasus exhibited anti-meta-static activity by stimulating the immune system. In this study, we fractionated glycoproteins (EN-SP) from the soluble protein layer (GF-AS) of A. senticasus and determined their basic chemical properties. We also investigated the anti-tumor and immunostimulating activities of the fractionated glycoprotein, EN-SP. We found that intravenous (i.v.) administration of GF-AS dramatically inhibited metastasis of colon26-M3.1 carcinoma cells to the lung in a dose-dependent manner. In vitro analysis showed GF-AS to enhance the proliferation of splenocytes. GF-AS also stimulated peritoneal macrophage, which was followed by the production of various cytokines such as IL-1$\beta$, TNF-$\alpha$, IL-12 and IFN-${\gamma}$. Furthermore, the production of these cytokines was partially blocked when peritoneal macrophage was cultured with the polyclonal antibodies against GF-AS. The depletion of NK cells by rabbit anti-asialo GM1 serum partly abolished the inhibitory effect of GF-AS on lung metastasis of colon26-M3.1 cells. Using gel filtration, EN-SP, an active glycoprotein fraction, is isolated from GF-AS. While both GF-AS and EN-SP stimulated the proliferatation of splenocytes of normal mice, EN-SP showed higher anti-metastatic activity and more potently stimulated the proliferation of splenocytes compared to GF-AS. These results suggest the use of EN-SP, the fractionated glycoprotein from A. senticasus, can be used as a therapeutical reagent to prevent or inhibit tumor metastasis.
Journal of The Korean Society of Clinical Toxicology
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v.17
no.2
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pp.66-78
/
2019
Purpose: Thallium (TI+) autometallography is often used for the imaging of neuronal metabolic activity in the rodent brain under various pathophysiologic conditions. The purpose of this study was to apply a thallium autometallographic technique to observe changes in neuronal activity in the forebrain of rats following acute carbon monoxide (CO) intoxication. Methods: In order to induce acute CO intoxication, adult Sprague-Dawley rats were exposed to 1100 ppm of CO for 40 minutes, followed by 3000 ppm of CO for 20 minutes. Animals were sacrificed at 30 minutes and 5 days after induction of acute CO intoxication for thallium autometallography. Immunohistochemical staining and toluidine blue staining were performed to observe cellular damage in the forebrain following intoxication. Results: Acute CO intoxication resulted in significant reduction of TI+ uptake in major forebrain structures, including the cortex, hippocampus, thalamus, and striatum. In the cortex and hippocampal CA1 area, marked reduction of TI+ uptake was observed in the cell bodies and dendrites of pyramidal neurons at 30 minutes following acute CO intoxication. There was also strong uptake of TI+ in astrocytes in the hippocampal CA3 area following acute CO intoxication. However, there were no significant histological findings of cell death and no reduction of NeuN (+) neuronal populations in the cortex and hippocampus at 5 days after acute CO intoxication. Conclusion: The results of this study suggest that thallium autometallography can be a new and useful technique for imaging functional changes in neural activity of the forebrain structure following mild to moderate CO intoxication.
Ha Heon-Seok;Kim Chang-Whe;Lim Young-Jun;Kim Myung-Joo
The Journal of Korean Academy of Prosthodontics
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v.44
no.3
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pp.343-355
/
2006
Statement of problem. The success of osseointegration can be enhanced with an implant that has improved surface characteristics. Anodic oxidation is one of the surface modifying method to achieve osseointegration. Voltage of anodic oxidation can change surface characteristics and cell activity Purpose. This study was performed to evaluate MG63 cell responses such as affinity, proliferation and to compare surface characteristics of anodic oxidized titanium in various voltage. Material and method. The disks for cell culture were fabricated from grade 3 commercially pure titanium,1 m in thickness and 12 mm in diameter. Surfaces of 4 different roughness were prepared. Group 1 had a machined surface, used as control. Group 2 was anodized under 220 V, group 3 was anodized under 300 V and group 4 was anodized under 320 V. The microtopography of specimens was observed by scanning electron microscope (JSM-840A, JEOL, Japan) and atomic force microscope(Autoprobe CP, Park Scientific Instrument, USA). The surface roughness was measured by confocal laser scanning microscope(Pascal, LSM5, Zeiss, Germany). The crystal structure of the titanium surface was analyzed with x-ray diffractometer(D8 advanced, Broker, Germany). MG63 osteoblast-like cells were cultured on these specimens. The cell morpholgy was observed by field emission electron microscope(Hitachi S-4700, Japan). The cell metabolic and proliferative activity was evaluated by MTT assay Results and conclusion. With in limitations of this in vitro study, the following conclusions were drawn. 1. In anodizing titanium surface, we could see pores which did not show in control group. In higher anodizing voltage, pore size was increased. 2. In anodizing titanium surface, we could see anatase. In higher anodizing voltage, thicker oxide layer increased crystallinity(anatase, anatase and rutile mixed). 3. MG63 cells showed more irregular, polarized and polygonal shape and developed more lamellipodi in anodizing group as voltage increased. 4. The activity of cells in MTT assay increased significantly in group 3 and 4 in comparison with group 1 and 2. However, there was no difference between group 3 and 4 at P<0.05. Proliferation of MG63 cells increased significantly in pore size($3-5.5{\mu}m$) of group 3 and 4 in comparison with in pore size($0.2-1{\mu}m$ ) of group 2.
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