Selenium is an essential micronutrient for normal body function and functions as an essential constituent of selenoproteins. This study was carried out to investigate effect of selenium on the formation of colonic aberrant crypt foci (ACF) and tumor formation in a mouse model. Five-week old ICR mice were acclimated for one week and fed different selenium diet (0.02, 0.1, and 0.5 ppm) for 12 weeks. Animals received three intraperitoneal injections of azoxymethane (10 mg/kg B.W. in saline for 3 weeks), followed by 2% dextran sodium sulfate in the drinking water for a week. There were four experimental groups, including a normal control group and three different selenium levels groups. After sacrifice, the total numbers of aberrant crypt (AC) and ACF were measured in the colonic mucosa after methylene blue staining. The number of tumors was noted for tumor incidence. Liver selenium concentration was measured using ICP-AES method. Gutathione peroxidase (GPx) activity was determined using a GPx assay kit in the liver and colon. TUNEL assay and proliferating cell nuclear antigen (PCNA) staining were performed to examine the cell apoptosis and cell proliferation, respectively. Immunohistochemistry of $\beta$-catenin was also performed on the mucous membrane tissue of colon. The activity of GPx in the liver and colon was decreased in the selenium-deficient diet group while it was increased in the selenium-overloaded diet group. Apoptotic positive cells were increased in the selenium-overloaded diet group but decreased in the selenium-deficient diet group. PCNA staining area was decreased in the selenium-overloaded diet group. In addition, the $\beta$-catenin protein level in the selenium-deficient diet group was increased but decreased in the selenium-overloaded diet group. These results indicate that dietary selenium might exert a modulating effect on colon cancer by inhibiting the development of ACF and colon tumor formation in this mouse model.
Purpose : Ginkgo biloba extract(GBE) is known to increase the peripheral blood circulation. This study was designed to evaluate the effect of GBE on the acute normal tissue radiation reaction. Materials and Methods : mice were divided into two groups, radiation alone and two doses GBE plus radiation, for both acute skin reaction and jejunal crypt assay. GBE was given i.p. one hour before irradiation with priming dose given one day earlier. Thirty to Fifty Gy for acute skin reaction and 11 to 14 Gy for jejunal crypt were irradiated to right hind leg and whole body, respectively. Results : Radiation doses($RD_{50}$) for Peak skin score of 2.0 were 44.2Gy (40.6-48.2Gy) for radiation alone and 44.4Gy(41.6-47.4Gy) for two doses GBE plus radiation, showing no effect of GBE on acute radiation skin damage. The numbers of regenerating jejunal crypts per circumference were also almost the same for each radiation dose level(p=0.57-0.94), and the mean lethal doses($D_o$) were 1.800y(1.57-2.09Gy) for radiation alone and 1.88Gy(1.65-2.18Gy) for two doses GBE plus radiation, indicating no effect of GBE on jejunal crypt cell survival after radiation. Conclusion : GBE doesn't increase acute normal tissue radiation reaction in this model system. As GBE was verified to enhance radiation effect on tumor, high therapeutic gain is expected when GBE is combined with radiation therapy.
Two experiments were conducted to investigate the in vitro ability of keratinase to hydrolyze soybean glycinin and ${\beta}$-conglycinin and to evaluate the in vivo effects of keratinase when included in corn-soybean diets with different levels of crude protein and fed to nursery pigs. In experiment 1, a saturated keratinase solution (1 ml) was added to two blank controls of either glycinin or ${\beta}$-conglycinin resulting in the hydrolysis of 94.74% glycinin and 88.89% ${\beta}$-conglycinin. In experiment 2, 190 pigs (8.3${\pm}$0.63 kg BW) were allotted to one of four treatments in a 2${\times}$2 factorial arrangement on the basis of body weight, and sex was balanced among the pens. The effects of crude protein (19 vs. 22%) and keratinase (0 vs. 0.05%) were studied. Each treatment was applied to six pens with seven (two pens) or eight pigs per pen. Pigs were fed the experimental diets for 21 d. Weight gain and feed conversion ratio were improved (p<0.05) with keratinase supplementation while feed intake was reduced (p<0.05). Keratinase supplementation increased (p<0.05) the apparent total tract digestibility of dry matter, energy, crude protein and phosphorus. Keratinase supplementation also increased n-butyric acid in the cecum and colon, lactobacilli and total anaerobe counts in the colon as well as the ratio of villus height to crypt depth in the ileum. Additionally, fecal score, ammonia nitrogen and branch chain volatile fatty acids in the colon, E. coli and total aerobe counts in the colon, crypt depth in the jejunum and ileum as well as serum interleukin-1 and interleukin-6 concentrations were also decreased (p<0.05) by keratinase supplementation. A reduction in dietary crude protein decreased (p<0.05) colon ammonia nitrogen concentration and cecal propionic acid and branch chain volatile fatty acid concentrations. In addition, cecal E. coli counts, colon total anaerobe counts, ileal crypt depth, and serum interleukin-1 and interleukin-6 concentrations were also decreased (p<0.05) with the reduction of dietary crude protein. With the exception of fecal scores, there were no significant interactions between crude protein and keratinase. This study provides evidence that dietary keratinase supplementation improved nursery pig performance by improving intestinal morphology and ecology, thus improving nutrient digestibility and alleviating the inflammatory response.
A total of 90 one-day-old male broilers (Ross 308) were randomly assigned to one of the three dietary treatments, each consisting of six replicates (5 broilers/cage). The dietary treatments were 1) control (CON: fresh clean water with no supplement); 2) low dose [LD: CON + 1.56% extractions from the wooden chips (EWC)] and 3) high dose (HD: CON + 12.5% EWC). Drinking water supplemented with EWC was provided using specifically designed individual nipple drinker units. Average daily water intake (ADWI), average daily gain (ADG), average daily feed intake (ADFI), and feed conversion ratio (FCR) were measured weekly for 21 days. One broiler from each cage was euthanized for measuring the visceral organ weights and collecting ileal tissue samples for ileal architecture analysis on day 21. Broilers assigned to the LD and HD watery groups showed higher ADWI than that in broilers consumed CON on day 7 (P<0.05). The broilers subjected to HD treatment showed a deeper crypt depth (P<0.05) than that in broilers subjected to LD and CON on day 21. Therefore, broilers consumed HD showed a lower (P<0.05) villus height:crypt depth ratio than that broilers consumed CON on day 21. Broilers provided drinking water containing any of the tested concentrations of EWC showed no effect (P > 0.05) on growth performance, ileal villus height, and visceral organ weights as compared with those in the CON from hatch to 21 days. In conclusion, broilers fed HD showed reduction in villus height:crypt depth ratio without impairing growth performance and visceral organ weights for the experimental period.
Aloe vera gel exhibits protective effects against insulin resistance as well as lipid-lowering and anti-diabetic effects. The anti-diabetic compounds in this gel were identified as Aloe-sterols. Aloe vera gel extract (AVGE) containing Aloe-sterols has recently been produced using a new procedure. We previously reported that AVGE reduced large-sized intestinal polyps in Apc-deficient Min mice fed a high fat diet (HFD), suggesting that Aloe vera gel may protect against colorectal cancer. In the present study, we examined the effects of Aloe vera gel powder (AVGP) and AVGE on azoxymethane-induced colorectal preneoplastic aberrant crypt foci (ACF) in mice fed a HFD. Male C57BL/6J mice were given a normal diet (ND), HFD, HFD containing 0.5% carboxymethyl cellulose solution, which was used as a solvent for AVGE (HFDC), HFD containing 3% or 1% AVGP, and HFDC containing 0.0125% (H-) or 0.00375% (L-) AVGE. The number of ACF was significantly lower in mice given 3% AVGP and H-AVGE than in those given HFD or HFDC alone. Moreover, 3% AVGP, H-AVGE and L-AVGE significantly decreased the mean Ki-67 labeling index, assessed as a measure of cell proliferation in the colonic mucosa. In addition, hepatic phase II enzyme glutathione S-transferase mRNA levels were higher in the H-AVGE group than in the HFDC group. These results suggest that both AVGP and AVGE may have chemopreventive effects on colorectal carcinogenesis under the HFD condition. Furthermore, the concentration of Aloe-sterols was similar between 3% AVGP and H-AVGE, suggesting that Aloe-sterols were the main active ingredients in this experiment.
The relationship between the intestinal histopathology and number and position of intraepithelial lymphocytes (IEL) was observed chronologically in the small intestine of rats experimentally infected with Echinostoma hortense. Sprague-Dawley rats were orally infected with 200 metacercariae obtained from Misgurnus anguillicaudatus. The rats each were sacrificed on the week 1, 2, 4, 6, 8 post-infection (PI) and samples of the intestine in the part of duodenum and jejunum were taken. The samples were stained with Hematoxylin-eosin and Giemsa. The intestinal histopathology was the severest after the week 1 PI and characterized by villous atrophy, crypt hyperplasia and decrease of villus/crypt(v/c) ratio, which continued until the week 8 Pl. The number of IEL dramatically decreased during the week 1 PI, but increased gradually thereafter with a slight decrease on the week 8 PI. In control rats, the great majority of the IEL were located at the basal region of the epithelium. During the early stage of infection, however, we found a considerable proportion of IEL to moved to the intermediate or apical regions of the epithelium. From the above results, it is sugested that the change of IEL number and position during the course of E. hortense infection should be closely related to the progression and recovery of the intestinal histopathology.
Obesity is an established risk factor for colorectal cancer. Pioglitazone is a peroxisome proliferator activated receptor$receptor{\gamma}$ ($PPAR{\gamma}$) agonist that induces differentiation in adipocytes and induces growth arrest and/or apoptosis in vitro in several cancer cell lines. In the present study, we investigated the effect of pioglitazone on the development of azoxymethane-induced colon aberrant crypt foci (ACF) in KK-$A^{\mathcal{Y}}$ obesity and diabetes model mice, and tried to clarify mechanisms by which the $PPAR{\gamma}$ ligand inhibits ACF development. Administration of 800 ppm pioglitazone reduced the number of colon ACF/mouse to 30% of those in untreated mice and improved hypertrophic changes of adipocytes in KK-$A^{\mathcal{Y}}$ mice with significant reduction of serum triglyceride and insulin levels. Moreover, mRNA levels of adipocytokines, such as leptin, monocyte chemoattractant protein-1 and plasminogen activator inhibitor-1, in the visceral fat were decreased. PCNA immunohistochemistry revealed that pioglitazone treatment suppressed cell proliferation in the colorectal epithelium with elevation of p27 and p53 gene expression. These results suggest that pioglitazone prevented obesity-associated colon carcinogenesis through improvement of dysregulated adipocytokine levels and high serum levels of triglyceride and insulin, and increase of p27 and p53 mRNA levels in the colorectal mucosa. These data indicate that pioglitazone warrants attention as a potential chemopreventive agent against obesity-associated colorectal cancer.
In order to clarify the effect of radiation on the mouse jejunal crypt cells by combined administration of administration and radiation and also to evaluate the enhancing effect of adriamycin, the authors performed this study by delivering single irradiation of 1,000 to 1,600 rad to the whole abdomen of mice by cobalt-60 teletherapy unit. In combination with adriyamycin treatment groups, the drug was administered as single dose of 10 mg/kg either 2 hours before or 4 hours after graded single dose,900 to 1,400 rad, of irradiation. The authors studied the quantitative changes of intestinal crypt cells by microcolony survival assay technique and the morphological changes of small intestinal villi by scanning electron microscope in mice following to combined therapy with adriamycin and irradiation, The average number of jejunal crypts per circumference was $130{\pm}16$ in control group. The mean lethal dose(Do) of each irradiation alone and combined therapy groups 2 hours before and 4 hours after irradiation, were 160, 170, and 170 rad in cell survival curves, respectively. The dose effect factor(DEF) of adriamycin in each groups of pre-irradiation and post-irradiation were 1.19 and 1.26, respectively. The conical shaped villi were noted on 1,200 rad in irradiation alone group and 1,000 rad in combined groups. For the proper clinical application we must be careful of the radiation injury to small bowel when the anticancer chemotherapy and radiation therapy to the abdomen and pelvic area are used as combined therapeutic modality.
Han, X.;Piao, X.S.;Zhang, H.Y.;Li, P.F.;Yi, J.Q.;Zhang, Q.;Li, P.
Asian-Australasian Journal of Animal Sciences
/
v.25
no.4
/
pp.569-576
/
2012
Two experiments were conducted to investigate the potential for Forsythia suspensa extract (FSE) to substitute for antibiotic in broiler chicken. First, a well-diffusion assay procedure and a 2-fold dilution method were used to determine the bacteriostatic activity of FSE on Escherichia coli K88, staphylococcus aureus, and salmonella was assayed. An inhibitory effect of FSE was observed on the growth of these bacteria. This effect seems to be dose depended, which disappeared after 25.00, 12.50, 1.56 mg/ml. Second, a 42-d trial with 252 broiler chickens (d 1, $38.7{\pm}1.1$ g BW) was conducted to evaluate the effect of dietary supplementation of FSE in broiler chicken. The feeding program consisted of a starter diet from d 1 to 21 and a finisher diet from d 22 to 42. Dietary treatments included were: i) NC: negative control fed a corn-soybean meal based diet; ii) PC: positive control group fed based diet with chlortetracycline; and iii) FC: a test group fed with 100 mg FSE/kg diet. In this study growth performance did not differ among treatments during the starter period. However, dietary supplemental chlortetracycline and FSE increased (p<0.05) average daily gain (ADG), average daily feed intake (ADFI) compared with NC during the finisher and overall phase. Apparent digestibility of calcium on d 21, digestibility of energy and calcium on d 42 of FC was greater (p<0.05) than NC. Moreover, cecal Escherichia coli counts for birds from FC were lower (p<0.05) than NC. Dietary FSE supplementation also improved (p<0.05) villus height and villus height to crypt depth ratios in both duodenum and ileum and decreased (p<0.05) crypt depth in the duodenum. Duodenum villus height and villus height to crypt depth ratio in both duodenum and ileum from the FC group were also greater (p<0.05). Serum growth hormone and IGF-1 were not influenced by different treatments. Apparently, FSE has the potential to substitute for antibiotic in broiler chicken.
Saenphoom, P.;Liang, J.B.;Ho, Y.W.;Loh, T.C.;Rosfarizan, M.
Asian-Australasian Journal of Animal Sciences
/
v.26
no.4
/
pp.537-544
/
2013
This study examined whether pre-treating palm kernel expeller (PKE) with exogenous enzyme would degrade its fiber content; thus improving its metabolizable energy (ME), growth performance, villus height and digesta viscosity in broiler chickens fed diets containing PKE. Our results showed that enzyme treatment decreased (p<0.05) hemicellulose and cellulose contents of PKE by 26.26 and 32.62%, respectively; and improved true ME (TME) and its nitrogen corrected value ($TME_n$) by 38% and 33%, respectively, compared to the raw sample. Average daily gain (ADG), feed intake and feed conversion ratio (FCR) of chickens fed on different dietary treatments in the grower period were not significantly different. Although there was no difference in feed intake (p>0.05) among treatment groups in the finisher period, ADG of chickens in the control (PKE-free diet) was higher (p<0.05) than in all treatment groups fed either 20 or 30% PKE, irrespective of with or without enzyme treatment. However, ADG of birds fed with 20% PKE was higher than those fed with 30% PKE. The FCR of chickens in the control was the lowest (2.20) but not significantly different from those fed 20% PKE diets while birds in the 30% PKE diets recorded higher (p>0.05) FCR. The intestinal villus height and crypt depth (duodenum, jejunum and ileum) were not different (p>0.05) among treatments except for duodenal crypt depth. The villus height and crypt depth of birds in enzyme treated PKE diets were higher (p<0.05) than those in the raw PKE groups. Viscosity of the intestinal digesta was not different (p>0.05) among treatments. Results of this study suggest that exogenous enzyme is effective in hydrolyzing the fiber (hemicellulose and cellulose) component and improved the ME values of PKE, however, the above positive effects were not reflected in the growth performance in broiler chickens fed the enzyme treated PKE compared to those received raw PKE. The results suggest that PKE can be included up to 5% in the grower diet and 20% in the finisher diet without any significant negative effect on FCR in broiler chickens.
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