• Biomolecules & Therapeutics
• /
• v.20 no.2
• /
• pp.177-182
• /
• 2012

Tetrazolium violet is a tetrazolium salt and has been proposed as an antitumor agent. In this study, we reported for the first time that tetrazolium violet not only inhibited human lung cancer A549 cell proliferation but also induced apoptosis and blocked cell cycle progression in the G1 phase. The results showed that tetrazolium violet significantly decreased the viability of A549 cells at $5-15{\mu}M$. Tetrazolium violet -induced apoptosis in A549 cells was confirmed by H33258 staining assay. In A549, tetrazolium violet blocked the progression of the cell cycle at G1 phase by inducing p53 expression and further up-regulating p21/WAF1 expression. In addition, an enhancement in Fas/APO-1 and its two forms of ligands, membrane-bound Fas ligand (mFasL) and soluble Fas ligand (sFasL), as well as caspase, were responsible for the apoptotic effect induced by tetrazolium violet. The conclusion of this study is that tetrazolium violet induced p53 expression which caused cell cycle arrest and apoptosis. These findings suggest that tetrazolium violet has strong potential for development as an agent for treatment lung cancer.

• Biomedical Science Letters
• /
• v.2 no.1
• /
• pp.49-55
• /
• 1996

The objective of this study was to develop a paper strip which could determine E. coli qualitatively and quantitatively in water, wastewater, drinks, or food. This paper strip method was a simple and rapid test method that determine E. coli by visual identification. In this study, nutrient culture media were formulated and characterized for optimum conditions. Paper strips were then prepared by impregnating into the media and dried at $60^\circ$C. The test procedure is quite simple to use. The paper strip was dipped into a sample, and excess sample was removed. The strip was then incubated at $37^\circ$C for 16 to 20 hours and the number of colonies on the strip was counted. The color of the colony spots produced by microorganisms varied depending on the media formulation. Violet-red spots were produced by E. coli. The test method was simple, rapid and no special laboratory equipment was necessary for visual identification. Therefore, this test method is applicable to on-site tests such as field tests or home tests. The paper strip method was compared with the standard agar plate method and Japanese commercial product. The method of the economical preparation of test strips was studied for production on industrial scale.

• The Journal of Korean Medicine
• /
• v.22 no.1
• /
• pp.10-21
• /
• 2001

Objective : This study was performed to investigate the protective effect of Stephania tetrandra(ST) against ischemic brain damage after a middle cerebral artery(MCA) occlusion. The effect was evaluated using histological tests, neurobehavioral tests, and biochemical tests. Methods : Rats(Sprague-Dawley) were divided into four groups : sham operated group, MCA occluded group, post MCA occlusion Stephania tetrandra administrated (7.6mg/l00g) group, and normal group. The MCA was occluded by intraluminal method. Stephania tetrandra was administrated orally twice at 1 and 4 hours after MCA occlusion. The neurobehavioral test was performed at 3, 6, 9 and 24 hours after MCA occlusion by posture reflex test and swimming behavioral test. All groups were sacrificed then. The brain tissues were stained with 2% triphenyl tetrazolium chloride(TTC) or 1 % cresyl violet solution, to examine infarct size, volume and cell number. Tumor necrosis $factor-{\alpha}$ level was measured from sera using Enzyme-Linked Immunoabsorbent Assay(ELISA). The mRNA expression level of inflammatory cytokines and related receptor type I and II, $IL-1{\beta}$, IL-6, and IL-10 6hours after MCA occlusion were also studied by reverse transcriptase polymerase chain reaction(RTPCR). Results : The results showed that : Stephania tetrandra (1) reduced infarct size and total infarct volume by 52.2% compared to the control group; (2) attenuated significantly in neuronal death, which was shown by a decrease in cell number(P<0.01) and size(P<0.01) in the boundary area of the infarction; (3) significantly reduced serum $TNF-{\alpha}$ level, and increased the mRNA level of IL-10 in the cortex region(P<0.01). However, there was no significant effect on motor deficit in swimming behavioral test. Conclusions : In conclusion, Stephania tetrandra has protective effects against ischemic brain damage at the early stage of ischemia.

• The Korea Journal of Herbology
• /
• v.26 no.3
• /
• pp.75-82
• /
• 2011

Objectives : In this study, we evaluated the effect of Chungganhaeju-hwan(CGHJH) on hydrogen peroxide($H_2O_2$)-induced and ethanol(EtOH)-induced neuronal damage in vitro and in vivo, respectively. Methods:We carried out the anti-oxidant effects of CGHJH against hydrogen peroxide($H_2O_2$)-induced toxicity in HT22 and PC12 cells using thiazolyl blue tetrazolium bromide. Then, to investigate the protective effect on CGHJH against EtOH-induced memory impairment and hippocampal cell damage in male ICR mice, we performed novel object recognition test(NORT), and analysed the brain tissues after immunohistochemistry and western blotting. Results:CGHJH showed protective effect from $H_2O_2$-induced cell toxicity at doses of $1\sim100{\mu}g$/mL in both HT22 and PC12 cells. CGHJH had also recovery effect from EtOH-induced memory impairment in ICR mice from NORT and it protected hippocampal cells against EtOH toxicity in the result of cresyl violet and NeuN immunoreactivity. Conclusion : These results demonstrate that CGHJH has protective effect in neuronal cells against $H_2O_2$ and EtOH toxicities and this effect could be a main role of recovery effect on EtOH-induced memory loss.

• The Journal of Korean Medicine
• /
• v.21 no.2
• /
• pp.25-36
• /
• 2000

Objectives : This study demonstrates the effects of Daejo-hwan on learning and memory impairment induced by L-NAME (75 mg/kg) treatment and on cerebral ischemic damage induced by middle cerebral artery (MCA) occlusion in rats. Methods : Daejo-hwan emulsion (73.3 mg/100 g/l ml) was administered to rats along a timed study schedule. The Moms water maze was used for learning and memory test of the rats. The MCA was occluded by using the intraluminal thread method. The brain slices were stained by 2 % triphenyl tetrazolium chloride (TTC) and 1 % cresyl violet solution. Infarct size, neuron cell number and size in penumbra was measured by using computer image analysis system. Results : 1. The escape latency of the Daejo-hwan treated group decreased significantly with respect to the control group. 2.The memory score of the Daejo-hwan treated group showed increase tendency, And the swimming distance was not different between the normal, the control, and the Daejo-hwan treated group. 3. The infarct size of the Daejo-hwan treated group decreased significantly with respect to the control group. 4. The total infarct volume of the Daejo-hwan treated group showed decrease tendency. And the brain edema index of the Daejo-hwan treated group decreased significantly with respect to the control group. 5. The neuron cell number and cell size in penumbra of the Daejo-hwan treated group increased significantly with respect to the control group. Conclusions : According to the above results, it is supposed that Daejo-hwan is clinically applicable to the vascular dementia.

• The Journal of Korean Medicine
• /
• v.21 no.2
• /
• pp.68-78
• /
• 2000

Objective : This research was performed to investigate protective effects of Sophora subprostrata, against ischemic brain damage after a middle cerebral artery(MCA) occlusion. The effect was estimated using histological test, neurobehavioural test, and biochemical test. Methods : Rats(Sprague-Dawley) were divided into four groups: Sham operated group, MCA occluded group, Sophora subprostrata administrated group after MCA occlusion, and Normal group. The MCA was occluded by intraluminal method. Sophora subprostrata was administrated orally twice(l and 4 hours) after middle cerebral artery occlusion. The neurobeavioural test was performed at 3 hours, 6 hours, 9 hours and 24 hours after the surgery by posture reflex test and swimming behavioural test. All groups were sacrificed at 24 hours after the surgery. The brain tissue was stained with 2% triphenyl tetrazolium chioride(TTC) or 1 % cresyl violet solution, to examine effect of Sophora subprostrata on ischemic brain tissue. The blood samples were obtained from the heart of rats. Tumor necrosis factor-a level was measured from sera using Enzyme-Linked Immunoabsorbent Assay(ELISA). Results : The results showed that (1) Sophora subprostrata reduced infarct size and total infarct volume by 54.8% compared to the control group, (2) that neuronal death, which was shown by decrease in cell number and size, was attenuated significantly in the boundary area of the infarction, (3) that serum $TNF-{\alpha}$ㆍlevel was reduced significantly, and finally, there was significant recovery of motor deficit at 3 hours after MCA occluded by Swimming behavioural test. Conclusions :In conclusion, Sophora subprostrata has protective effects against ischemic brain damage at the early stage of ischemia.

• Restorative Dentistry and Endodontics
• /
• v.45 no.4
• /
• pp.47.1-47.11
• /
• 2020

Objectives: The aim of this study was to assess the physicochemical properties, cytotoxicity and penetration into dentinal tubules of ChlorCid^TM Surf (3% sodium hypochlorite [NaOCl] with surfactant) in comparison to ChlorCid^TM (3% NaOCl without surfactant). Materials and Methods: The physicochemical properties evaluated were pH, surface tension, free available chlorine (FAC) and contact angle. Cytotoxicity was evaluated in L929 fibroblasts exposed to the solutions by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and neutral red assays. Assessment of penetration into dentinal tubules was performed by staining single-rooted permanent human teeth with crystal violet (n = 9), which were irrigated with the solutions and analyzed in cervical, middle and apical segments. Data were analyzed by one-way analysis of variance (ANOVA) and Tukey's post-test, 2-way ANOVA and Bonferroni's post-test or t-test (α = 0.05). Results: ChlorCid^TM Surf and ChlorCid^TM FAC values were close to those indicated by the manufacturer. ChlorCid^TM Surf showed lower surface tension and contact angle on dentin, and higher pH than ChlorCid^TM (p < 0.05). The penetration of ChlorCid^TM Surf was higher in cervical and middle segments, compared with ChlorCid^TM (p < 0.05). There was no difference in irrigant cytotoxicity (p > 0.05). Conclusions: ChlorCid^TM Surf showed lower surface tension, lower contact angle on root canal dentin, higher penetration into dentinal tubules and more alkaline pH, compared with ChlorCid^TM. However, both solutions showed similar cytotoxicity and FAC content.

• The Journal of Korean Medicine
• /
• v.26 no.2 s.62
• /
• pp.217-230
• /
• 2005

Objectives: Chungpaesagan-tang (CPSGT), which is frequently used for treating patients of cerebrovascular disease, has not been reported by clinical doctors concerning the effect of neuronal aptosis caused by brain ischemia. To study the effect of CPSGT on focal cerebral ischemia in normal and diabetic rats and SHR, focal cerebral ischemia was induced by transient MCAO, and after onset CPSGT was administrated. Methods: Rats (Sprague-Dawley) were divided into four groups: sham-operated group, MCA-occluded group, CPSGT­administrated group after MCA occlusion, and normal group. The MCA was occluded by intraluminal method. CPSGT was administrated orally twice (l and 4 hours) after middle cerebral artery occlusion. All groups were sacrificed at 24 hours after the surgery. The brain tissue Was stained with $2\%$ triphenyl tetrazolium chloride (TTC) or $1\%$ cresyl violet solution, to examine effect of CPSGT on ischemic brain tissue. The blood samples were obtained from the heart.~. Tumor necrosis $factor-\alpha$ level and interleukin-6 level of serum was measured from sera using enzyme-linked immunoabsorbent assay (ELISA). Then changes of immunohistochemical expression of $TNF-\alpha$ in ischemic damaged areas were observed. Results: In NC+MCAO+CP and DM+MCAO+CP, CPSGT significantly (p<0.01) decreased the number of neuron cells compared to the control group. CPSGT markedly reduced (p<0.01) the infarct size of the forebrain in distance from the interaural line on cerebral ischemia in diabetic rats. CPSGT significantly reduced the $TNF-\alpha$ expression in penumbra region of damaged hemisphere in diabetic rats. Conclusions: CPSGT had a protective effect on cerebral ischemia in SD rats, especially in diabetic rats compared with normal SD rats.

• The Journal of Internal Korean Medicine
• /
• v.34 no.2
• /
• pp.178-191
• /
• 2013

Objectives : In this study, we made an effort to investigate the protective mechanism of Ukgan-san (UGS) extracts on hypoxia-induced C6 glial cell death. Methods : The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were analysed with microscope after staining with crystal violet (CV). Reactive oxygen species (ROS) formation was assessed by flow cytometer after staining with 2'7'-dichlorofluorescein diacetate (DCF-DA). We also analyzed expression of hypoxia-inducible factor-1 alpha (HIF-$1{\alpha}$) and p53, processing of procaspase-3 and procyclic acidic repetitive protein (PARP) by western blot method. Results : We estimated the elevated cell viability by UGS extract on $CoCl_2$-induced C6 glial cells. UGS attenuated $CoCl_2$-induced ROS formation in C6 glial cells and also showed a protective activity compared to antioxidants and exhibited abrogation of LDH-released by $CoCl_2$. UGS suppressed the typical apoptotic cell death markers, caspase-3 and PARP activation. UGS inhibited $CoCl_2$-induced HIF-1${\alpha}$ expression which is known as a major regulator for hypoxia-induced cell death, and suppressed p53 expression. Conclusions : These results suggest that UGS extract contains protective constituents for hypoxia-induced C6 glial cell death.

• Microbiology and Biotechnology Letters
• /
• v.16 no.6
• /
• pp.517-521
• /
• 1988

This study focused on the psychrotrophic post-pasteurization contamination of fluid milk pro-ducts which were processed by HTST system. Pasteurized line samples and container samples of each fluid milk product (whole milk and skim milk) were taken in a large fluid milk plant. tine samples were collected through nine and five different sampling locations for whole milk and skim milk products, respectively. Each sample was subjected to preliminary incubation (PI) at 21$^{\circ}C$ for 16h followed by standard plate count (SPC) and crystal violet tetrazolium agar count (CVT). Flavor, SPC, and psychrotrophic bacteria count (PBC) were determined after 7 d at 7.2$^{\circ}C$. In addition, ten sequential container samples (packaged in 1000ml paperboard containers) were taken from a filler at the beginning of each product run. These samples were used for PI followed by SPC and CVT. In addition, flavor evaluations, SPC and PBC tests were conducted after 7,10, and 14 d at 7.2$^{\circ}C$. The mean PI-CVT values for the line samples showed differences depending on the location. There was major contamination between pasteurized storage tank and the filler. The PI-CVT counts for each container sample were negatively correlated with flayer scores at 10 and 140. There were good correlations among PI-CVT values of line samples and the percentage of total container samples with acceptable flavor after 10d.