Salim, Elsayed I;Hegazi, Mona M;Kang, Jin Seok;Helmy, Hager M
Asian Pacific Journal of Cancer Prevention
/
v.17
no.3
/
pp.1023-1035
/
2016
The purpose of this study was to investigate the role of colon cancer stem cells (CSCs) during chemically-induced rat multi-step colon carcinogenesis with or without the treatment with a specific cyclooxygenase-2 inhibitor drug (celecoxib). Two experiments were performed, the first, a short term 12 week colon carcinogenesis bioassay in which only surrogate markers for colon cancer, aberrant crypt foci (ACF) lesions, were formed. The other experiment was a medium term colon cancer rat assay in which tumors had developed after 32 weeks. Treatment with celecoxib lowered the numbers of ACF, as well as the tumor volumes and multiplicities after 32 weeks. Immunohistochemical proliferating cell nuclear antigen (PCNA) labeling indexes LI (%) were downregulated after treatment by celecoxib. Also different cell surface antigens known to associate with CSCs such as the epithelial cell adhesion molecule (EpCAM), CD44 and CD133 were compared between the two experiments and showed differential expression patterns depending on the stage of carcinogenesis and treatment with celecoxib. Flow cytometric analysis demonstrated that the numbers of CD133 cells were increased in the colonic epithelium after 12 weeks while those of CD44 but not CD133 cells were increased after 32 weeks. Moreover, aldehyde dehydrogenase-1 activity levels in the colonic epithelium (a known CSC marker) detected by ELISA assay were found down-regulated after 12 weeks, but were up-regulated after 32 weeks. The data have also shown that the protective effect of celecoxib on these specific markers and populations of CSCs and on other molecular processes such as apoptosis targeted by this drug may vary depending on the genetic and phenotypic stages of carcinogenesis. Therefore, uncovering these distinction roles of CSCs during different phases of carcinogenesis and during specific treatment could be useful for targeted therapy.
This experiment was performed to identify and isolate a growth inhibiting compound from Aralia continentalis. In order to isolate the growth inhibiting compound from Aralia continentalis the bioassay test of lettuce seed germination and rice seedling growth were used. Through these bioassays the growth inhibiting compound which was spotted at $R_f$ 0.51 on Tlc was isolated. This compound inhibited the lettuce growth by 79% at the concentration of 1000ppm. When sprayed with $FeCl_3$ reagent, it developed a bule spot. It had UV-absorbance at 217 nm and 342 nm, and $OH^-$ of $3600cm^{-1}$, C=O of $1700cm^{-1}$, C=C of $1600cm^{-1}$, and C-O of $1200cm^{-1}$ on IR spectrum. Through HPLC analysis this compound was identified as a ferulic acid ($C_{10}H_{10}O_4$) having 25 min. retention time.
Recently endogenous digitalis-like substances were found in the blood of various cardiovascular diseases and they have been considered one of the causes of evoking hypertension. However, the mechanism of endogenous digitalis-like substances-induced hypertension is not clarified yet. Therefore, the effects of Na-K pump inhibition on the contractility of vascular smooth muscle[conduit and resistant artery were investigated, using organ bath and bioassay experiment. Aortic and carotid arterial rings[conduit artery and the branches of brachial and superior mesenteric artery[resistant artery were used to find the effect of Na-K pump inhibition. The results obtained were as followes;The magnitudes of contractions induced by norepinephrine, serotonin, or acetylcholine in all these arteries were significantly increased by the inhibition of Na-K pump. The increased contractile responses to these agonists, especially to serotonin, were much more prominant in resistant arteries. Nitroprusside-induced relaxations were attenuated by Na-K pump inhibition and there were no significant differences in the effects of Na-K pump inhibition on nitroprusside-induced relaxations of these blood vessels. Endothelium-dependent relaxation was suppressed by the inhibition of Na-K pump, especially by the administration of ouabain, and this inhibitory effect was much more prominent in the branches of superior mesenteric artery, compared with other arteries. In the branches of superior mesenteric arteries, endothelium-dependent relaxation was completely blocked by ouabain. The release of EDRF was partially suppressed by Na-K pump inhibition.From the above results, it is suggested that the hypertension due to the increase in vascular resistance can be evoked by the inhibition of Na-K pump and endogenous digitalis-like substances induce hypertension through this mechanism.
Interior finishing materials using noncombustible were regulated by the building codes to prevent the spread of fire and protect occupants. The average deed of stopping time of experimental mouse exposing combustion gas were measured by KS F 2271 gas toxicity test. At that time, The average deed of stopping time under 9 minutes were judged a inconsistence. This experiment method has limit to find out a cause of toxicity effect factor. In this study, particle size analysis were performed for investigate a major factor.
The nitrofen adsorption on several clay minerals was determined by sludge method to know the effect of clay minerals on the nitrofen activity. The bioassay was conducted with wheat seedlings to study the influence of the adsorbed nitrofen on the nitrofen activity. It is apparant that a four hours shaking was enough to reach the equilibrium concentration. The more the amount of sample, the more nitrofen was adsorbed by clay minerals, whereas the more nitrofen adsorption per unit gram of sample was observed at a 200 mg addition than a 400 mg in the same nitrofen solution. A little amount of nitrofen was adsorbed on Ca-zeolite or Ca-kaolinte, and much more nitrofen was adsorbed on Na-montmorillonite than the other clay minerals in the experiment. Little effect of pH on the adsorption would be attributed to physical adsorption between nitrofen molecule and clay surface. Na-and Ca-montmorillonite were the most effective in reducing the phytotoxicity of nitrofen to the growth of wheat seedlings among clay minerals which nitrofen was added to the growth media.
To verify allelopathic effects, seed germination and seedling growth test, chemical analysis and bioassay of selected species were carried out with naturally occurring chemicals of Artemisia capillaris. Seed germination ratio of Calamagrostis arundinacea. Youngia denticulata and Lactuca indica var. laciniata showed decrease in proportion to increase in aqueous extracts concentration of A. capillaris. while that of Cosmos bipinnatus and Leonurus sibiricus did not. However, dry weight growth of selected species treated with the same extracts as the above experiment was inhibited remarkably compared to the germination test. In the test at different concentrations of essential oil from A. capillaris, seedling growth of A. princeps var. orientalis and Plantago asiatica was suppressed according to the concentration of the essential oil, and root growth of the selected species was more inhibitory than that of shoot growth. Thirty-six chemical compounds were identified from A. capillaris plant by gas chromatography. Seven compounds out of 36 were bioassayed, and terpinen-4-ol was the most toxic among the tested substances.
This study was carried out to know the Trifluralin adsorption experiment was determined with slurry method and the bioassay was conducted with barley seedlings. The adsorption of Trifluralin on clay minerals nearly was reached to eqilibrium concentration by shaking for 18 hours. The more the amount of clay minerals, the more Trifluralin was adsorbed by clay minerals. However the amount of Trifluralin adsorption per gram of clay minerals was reduced. The amount of Trifluralin adsorbed was higher on Montmorillonite than Zeolite and Kaolinite. And Kd value of Trifluralin was the greatest on the Montmorillonite.
Acute toxicity of crude oil (WSF) on the mortality and respiration rates of Neomysis awatschensis was examined. This experiment was conducted by static and short-term bioassay procedure. In lethal test, the test animals were exposured to 8 different concentrations to determine $LC_{50}$ value (median lethal concentration). The concentrations of total hydrocarbon of 96-hr $LC_{50}$ value at $14^{\circ}C\;and\;20^{\circ}C$ were 1.01 ppm and 0.78 ppm, respectively. $LT_{50}$ (the median lethal time) also was determined. The $LT_{50}$ of 0.56 ppm was found within 100 hours, while the $LT_{50}$ of 5.6 ppm was 21 hours at $14^{\circ}C$. At $20^{\circ}C$, the $LT_{50}$ values of 0.56 ppm and 5.60 ppm were 95 hours and 17 hours, respectively. There was little difference between two temperature experiments. The effect of WSF on respiration rate was more sensitive than that on mortality, but no considerable difference was shown between different concentrations in this experiment. The results of these experiments indicated that relatively low concentration of dissolved crude oil fraction can impact on small crustacean in the marine ecosystem.
Inflammatory process leads to the well-known mucosal damage and therefore a further disturbance of the epithelial barrier function, resulting abnormal intestinal wall function, even further accelerating the inflammatory process[1]. Despite of the records, etiology and pathogenesis of IBD remain rather unclear. There are many studies over the past couple of years have led to great advanced in understanding the inflammatory bowel disease(IBD) and their underlying pathophysiologic mechanisms. From the current understanding, it is likely that chronic inflammation in IBD is due to aggressive cellular immune responses including increased serum concentrations of different cytokines. Therefore, targeted molecules can be specifically eliminated in their expression directly on the transcriptional level. Interesting therapeutic trials are expected against adhesion molecules and pro-inflammatory cytokines such as TNF-${\alpha}$. The future development of immune therapies in IBD therefore holds great promises for better treatment modalities of IBD but will also open important new insights into a further understanding of inflammation pathophysiology. Treatment of cytokine inhibitors such as Immunex(Enbrel) and J&J/Centocor(Remicade) which are mouse-derived monoclonal antibodies have been shown in several studies to modulate the symptoms of patients, however, theses TNF inhibitors also have an adverse effect immune-related problems and also are costly and must be administered by injection. Because of the eventual development of unwanted side effects, these two products are used in only a select patient population. The present study was performed to elucidate the ability of TNF-${\alpha}$ antibodies produced in sheep colostrums to neutralize TNF-${\alpha}$ action in a cell-based bioassay and in a small animal model of intestinal inflammation. In vitro study, inhibitory effect of anti-TNF-${\alpha}$ antibody from the sheep was determined by cell bioassay. The antibody from the sheep at 1 in 10,000 dilution was able to completely inhibit TNF-${\alpha}$ activity in the cell bioassay. The antibodies from the same sheep, but different milkings, exhibited some variability in inhibition of TNF-${\alpha}$ activity, but were all greater than the control sample. In vivo study, the degree of inflammation was severe to experiment, despite of the initial pilot trial, main trial 1 was unable to figure out of any effect of antibody to reduce the impact of PAF and LPS. Main rat trial 2 resulted no significant symptoms like characteristic acute diarrhea and weight loss of colitis. This study suggested that colostrums from sheep immunized against TNF-${\alpha}$ significantly inhibited TNF-${\alpha}$ bioactivity in the cell based assay. And the higher than anticipated variability in the two animal models precluded assessment of the ability of antibody to prevent TNF-${\alpha}$ induced intestinal damage in the intact animal. Further study will require to find out an alternative animal model, which is more acceptable to test anti-TNF-${\alpha}$ IgA therapy for reducing the impact of inflammation on gut dysfunction. And subsequent pre-clinical and clinical testing also need generation of more antibody as current supplies are low.
To investigate the modifying effect of Kwao Kreu, Pueraria mirifica (PM), we performed two kind of studies which are the non-surgical medium-term carcinogenicity study and the modulation of gap junctional intercellular communication study. The first study, a non-surgical medium-term carcinogenicity bioassay was done to investigate the modifying effect of Kwao Keru, Pueyaria mirifca (PH), a rejuvenating folk medicine from Thailand, on the male F344 rat liver. Specific pathogen free, male 6-week-old F3444 rats were divided into ten groups. To induce hepatocarcinogenesis, those in all groups were given a single i.p. injection of DEN (200 mg/kg) and were received two i.p. injection of DGA (300 mg/kg) at the ends of weeks 2 and 5. Rats of group 3-6 were given sodium phenobarbital (PB 0.05% in drink). A diet containing 10 mg/kg PM was given to group 2 during the post-initiation phase and to groups 4 and 5 during promotion and initiation phase, respectively. Group 6 was given the experimental diet alone throughout the experiment (8 weeks). Rats of group 7, 8, 9 and 10 were fed 1000 mg/kg PH in the same manner as group 2, 4, 5 and 6. All animals were sacrificed at 8 weeks after DEN administration. Result of the immunohistochemical staining of the glutathione S-transferase placental form (GST-p) indicated that the numbers and areas of the preneoplastic leisions were not significantly changed in all PM treatment group comparing to control group. Also the numbers and areas of GST-p positive foci among group 7, 8, 9 and 10 were not significantly changed in comparing to control group. To study the effect of PM on the modulation of gap junctional intercellular communication, the present study was performed scrape-loading dye transfer (SL/DT) assay in human keratinocytes. The results showed that PM could not modulate GJIC. These results indicate that Pueraria mirifica may have no carcinogenic effects on experimental hepatocarcinogenesis in rats and gap junctional intercellular communication in human keratinocyte.
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