Expression of invasion/metastasis suppressor, E-cadherin, is reduced in many types of human carcinomas. Although somatic and germline mutations in the CDH1, which encodes the human E-cadherin, have frequently been reported in cases with diffuse gastric and lobular breast cancers, irreversible genetic inactivations are rare in other human carcinomas. Recently, it has been well documented that some genes in human cancers may be inactivated by altered CpG methylation. Herein, we determined the expression and methylation status of E-cadherin in oral squamous cell carcinoma(SCC) by immunohistochemistry and methylation-specific PCR. The expression of E-cadherin was significantly higher in the well-differentiated oral SCCs than the moderately or poorly differentiated ones. None of eight tested benign epithelial hyperplasias showed aberrant methylation, whereas five of 12 oral squamous cell carcinomas showed aberrant methylation. When we compared E-cadherin expression with methylation status, oral SCCs with normal methylation showed a higher expression of E-cadherin than those with methylation. These findings suggest that aberrant CpG methylation of CDH1 promoter region is closely associated with transcriptional inactivation and might be involved in tumor progression of the oral mucosa.
Objective: The effects of maternal undernutrition during midgestation on muscle fiber histology, myosin heavy chain (MyHC) expression, methylation modification of myogenic factors, and the mammalian target of rapamycin (mTOR) signaling pathway in the skeletal muscles of prenatal and postnatal goats were examined. Methods: Twenty-four pregnant goats were assigned to a control (100% of the nutrients requirement, n = 12) or a restricted group (60% of the nutrients requirement, n = 12) between 45 and 100 days of gestation. Descendants were harvested at day 100 of gestation and at day 90 after birth to collect the femoris muscle tissue. Results: Maternal undernutrition increased (p<0.05) the fiber area of the vastus muscle in the fetuses and enhanced (p<0.01) the proportions of MyHCI and MyHCIIA fibers in offspring, while the proportion of MyHCIIX fibers was decreased (p<0.01). DNA methylation at the +530 cytosine-guanine dinucleotide (CpG) site of the myogenic factor 5 (MYF5) promoter in restricted fetuses was increased (p<0.05), but the methylation of the MYF5 gene at the +274,280 CpG site and of the myogenic differentiation (MYOD) gene at the +252 CpG site in restricted kids was reduced (p<0.05). mTOR protein signals were down-regulated (p<0.05) in the restricted offspring. Conclusion: Maternal undernutrition altered the muscle fiber type in offspring, but its relationship with methylation in the promoter regions of myogenic genes needs to be elucidated.
cDNAs complementary to the 3'-terminal regions of two potyvirus genomes were cloned and sequenced. The clone G7 contains one open reading frame (ORF) of 1,338 nucleotides and a 3' untranslated region (3'-UTR) of 403 nucleotides at the 3'-end excluding the 3'end poly(A) tail. The putative viral coat protein (CP) shows 55%-92% amino acid sequence homology to those of Allium potyviruses. The genome size of the virus was analyzed to be about 9.0 kb by Northern blot analysis. Five cDNA clones were screened out using GPV2 oligonucleotide as a probe. One of these clones, DEA72, which has a longest cDNA insert, contains one ORF of 1,459 nucleotides and a 3'-UTR of 590 nucleotides at the 3'-end excluding the 3'-end poly(A) tail. The putative viral CP shows 57%-88% amino acid sequence homologies to those of Allium potyviruses. The genome size of the virus was analyzed to be about 9.6 kb by Northern blot analysis. The results of immunoblot and Northern blot analyses suggest that almost all of the tested garlic plants showing mosaic or streak symptoms are infected with DEA72-potyvirus in variable degrees but rarely infected with G7-potyvirus in variable degrees but rarely infected with DEA72-potyvirus in variable degrees but rarely infected with G7-potyvirus. Immunoelectron microscopy using anti-DEA72 CP antibody shows that this potyvirus is about 750 nm long and flexuous rod shaped.
Phuc, Bui Huy Nhu;Lindberg, Jan Erik;Ogle, Brian;Thomke, Sigvard
Asian-Australasian Journal of Animal Sciences
/
v.14
no.7
/
pp.986-993
/
2001
In balance experiments with rats either 25 or 50% of the casein protein in the control diet was replaced with one of the following eight sun-dried tropical biomass products: water spinach plants (WS) (Ipomoea aquatica Forsk), leucaena leaves (LL) (Leuceana leucocephala), duckweed plants (DW) (Lemma minor L.), groundnut foliage (OF) (Arachis hypogaea L.), trichantera leaves (Tric) (Trichantera gigantea), indicago leaves (Ind) (Indigofera hirsuta), mungbean foliage (Mb) (Phaseolus aureus), and cassava leaves (CL) (Manihot esculenta Crantz). The experiment included 102 rats with six individuals per treatment group. In three of the 16 biomass treatment groups, feed intake and weight gain of the rats were unacceptably low, and therefore they were excluded from the statistical evaluation, The crude protein (CP) content of the biomass products varied between 20.9% (Tric) and 33.2% (DW), whereas the content of NDF varied between 18.5% (Ind) and 32.2% (DW) of dry matter (DM). The total content of essential amino acids (g/16 g N) was comparable with that of alfalfa meal, except for GF and Tric, which were inferior. Between plant species, differences in dietary digestibility of organic matter (dOM) and CP (dCP) were observed (p<0.001). Also, the replacement level negatively influenced dOM and dCP (p<0.001). The lowest values for dOM (p<0.001) were observed for diets including biomass products with the highest content of NDF (OF, Tric, Mb, LL). Digestibility of CP was negatively affected by level of protein replacement. Significant (p<0.001) differences were found in N-retention and biological value among diets with different biomass products. The most favourable overall results were obtained for DW, WS and CL. The main factors affecting the nutritive value of the diets tested were their NDF content, dCP and AA profile of the biomass. Also antinutritive component(s) may have influenced the process of digestion and metabolism of some of the biomass products.
Kim, Sung Hwan;Lee, In Chul;Ko, Je Won;Moon, Changjong;Kim, Sung Ho;Shin, In Sik;Seo, Young Won;Kim, Hyoung Chin;Kim, Jong Choon
Biomolecules & Therapeutics
/
v.23
no.2
/
pp.180-188
/
2015
This study investigated the possible effects and molecular mechanisms of diallyl disulfide (DADS) against cyclophosphamide (CP)-induced hemorrhagic cystitis (HC) in rats. Inflammation response was assessed by histopathology and serum cytokines levels. We determined the protein expressions of nuclear transcription factor kappa-B (NF-${\kappa}B$), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), oxidative stress, urinary nitrite-nitrate, malondialdehyde (MDA), and 8-hydroxy-2'-deoxyguanosine (8-OHdG). Finally, we studied the involvement of mitogen-activated protein kinases (MAPKs) signaling in the protective effects of DADS against CP-induced HC. CP treatment caused a HC which was evidenced by an increase in histopathological changes, proinflammatory cytokines levels, urinary nitrite-nitrate level, and the protein expression of NF-${\kappa}B$, COX-2, iNOS, TNF-${\alpha}$, p-c-Jun N-terminal kinase (JNK), and p-extracellular signal regulated kinase (ERK). The significant decreases in glutathione content and glutathione-S-transferase and glutathione reductase activities, and the significant increase in MDA content and urinary MDA and 8-OHdG levels indicated that CP-induced bladder injury was mediated through oxidative DNA damage. In contrast, DADS pretreatment attenuated CP-induced HC, including histopathological lesion, serum cytokines levels, oxidative damage, and urinary oxidative DNA damage. DADS also caused significantly decreased the protein expressions of NF-${\kappa}B$, COX-2, iNOS, TNF-${\alpha}$, p-JNK, and p-ERK. These results indicate that DADS prevents CP-induced HC and that the protective effects of DADS may be due to its ability to regulate proinflammatory cytokines production by inhibition of NF-${\kappa}B$ and MAPKs expressions, and its potent anti-oxidative capability through reduction of oxidative DNA damage in the bladder.
The physicochemical properties of tagatose, a low calorie sweetener, was investigated. Rheological property of tagatose solution was found to be Bingham fluid. As the concentration of tagatose increased from 10 to 50% at $25^{\circ}C$, the viscosity increased from 1.65 to 5.14 cp. When the temperature of 40% tagatose solution increased from 15 to $55^{\circ}C$, the viscosity decreased from 4.59 to 2.33cp. The melting onset temperature and endothermic enthalpy of tagatose were $130.4^{\circ}C$ and -202.3 J/g, respectively, which were obtained from the analysis of differential scanning calorimetry. Tagatose showed higher water absorption than sucrose under $85{\sim}100%$ of relative humidity. Tagatose was less soluble than sucrose at $20{\sim}70^{\circ}C$. Water activity of tagatose in 60% concentration was 0.892, which was lower than 0.957 of sucrose solution. Tagatose solution adjusted from pH 2 to pH 12 was stable after 3 days. Amount of tagatose was not changed after heat treatment at $154^{\circ}C$ for 4 hours. But a browning reaction was found and absorbance of a tagatose solution increased with heat treatment.
Simultaneous reduction of Cr(VI) and degradation of phenol was observed in batch and bench-scale continuous stirred tank reactors using Rhodococcus sp. CP01 isolated from leachate. The strain CP01, which was capable of utilizing phenol as a sole source of carbon and energy, completely reduced added hexavalent chromium (0.25 mM) to its trivalent form during 60 hr batch assay under optimal conditions (pH 7.0 and 1,000 mg/L of phenol concentration). The rates of Cr(VI) reduction and phenol degradation were estimated as 4.17 $\mu$M Cr(VI) and 38.4 mg phenol.$L^{-1}{\cdot}hr^{-1}$, respectively. The continuous culture experiment was conducted for 46 days using synthetic feed containing different levels of chromate (0.0625 to 0.25 mM) and phenol(1,000 to 4,000 mg/L). With a hydraulic retention time of 100 hr, Cr(VI) reduction efficiency was mostly 100% for influent Cr(VI) and phenol concentrations of 0.125 mM and 3,000 mg/L, respectively. During quasi-steady-state operation, specific rate of Cr(VI) reduction was calculated as 0.34 mg Cr(VI).g $protein^{-1}{\cdot}hr^{-1}$ which was comparable to reported values obtained by using glucose as growth substrate. The results suggest the potential application of biological treatment for detoxification of wastewater contaminated simultaneously with Cr(VI) and pheonol.
Chang, Yoon Soo;Lee, Ho-Young;Kim, Young Sam;Kim, Hyung Jung;Chang, Joon;Ahn, Chul Min;Kim, Sung Kyu;Kim, Se Kyu
Tuberculosis and Respiratory Diseases
/
v.56
no.5
/
pp.465-484
/
2004
Background : Insulin-like growth factor (IGF)-binding protein-3 (IGFBP-3) inhibits the proliferation of non-small cell lung cancer (NSCLC) cells by inducing apoptosis. Methods : In this study, we investigated whether hypermethylation of IGFBP-3 promoter play an important role in the loss of IGFBP-3 expression in NSCLC. We also studied the mechanisms that mediate the silencing of IGFBP-3 expression in the cell lines which have hypermethylated IGFBP-3 promoter. Results : The IGFBP-3 promoter has hypermethylation in 7 of 15 (46.7%) NSCLC cell lines and 16 (69.7%) of 23, 7 (77.8%) of 9, 4 (80%) of 5, 4 (66.7 %) of 6, and 6 (100%) of 6 tumor specimens from patients with stage I, II, IIIA, IIIB, and IV NSCLC, respectively. The methylation status correlated with the level of protein and mRNA in NSCLC cell lines. Expression of IGFBP-3 was restored by the demethylating agent 5'-aza-2'-deoxycytidine (5'-aza-dC) in a subset of NSCLC cell lines. The Sp-1/ Sp-3 binding element in the IGFBP-3 promoter, important for promoter activity, was methylated in the NSCLC cell lines which have reduced IGFBP-3 expression and the methylation of this element suppressed the binding of the Sp-1 transcription factor. A ChIP assay showed that the methylation status of the IGFBP-3 promoter influenced the binding of Sp-1, methyl-CpG binding protein-2 (MeCP2), and histone deacetylase (HDAC) to Sp-1/Sp-3 binding element, which were reversed by by 5'-aza-dC. In vitro methylation of the IGFBP-3 promoter containing the Sp-1/Sp-3 binding element significantly reduced promoter activity, which was further suppressed by the overexpression of MeCP2. This reduction in activity was rescued by 5'-aza-dC. Conclusion : These findings indicate that hypermethylation of the IGFBP-3 promoter is one mechanism by which IGFBP-3 expression is silenced and MeCP2, with recruitment of HDAC, may play a role in silencing of IGFBP-3 expression. The frequency of this abnormality is also associated with advanced stages among the patients with NSCLC, suggesting that IGFBP-3 plays an important role in lung carcinogenesis/progression and that the promoter methylation status of IGFBP-3 may be a marker for early molecular detection and/or for monitoring chemoprevention efforts.
Xande, X.;Despois, E.;Giorgi, M.;Gourdinegi, J.L.;Archimedee, H.;Renaudeau, D.
Asian-Australasian Journal of Animal Sciences
/
v.22
no.1
/
pp.90-98
/
2009
The aim of this study was to evaluate the effect of a milling by-product diet and two sugar cane diets on the local Creole pig breed (CR). A total of 48 CR pigs (24 females and 24 castrated males) were randomly assigned to four different groups of 12 animals. Pigs were allotted to one of 4 dietary treatments: fed with a control soya-bean meal-corn diet containing 19.1% crude protein (CP) and 15.4 MJ DE/kg (diet 1), with an experimental milling by-product diet (soya-bean meal and wheat by-products) containing 19.4% CP and 13.0 MJ DE/kg (diet 2), with ground cane stalks (GCS) or with fresh sugar cane juice (SCJ). Both GCS and SCJ were supplemented with soya-bean meal complement (400 g/d of a 48.7% CP and 16.1 MJ DE/kg diet) in order to obtain diets 3 and 4, respectively. Pigs were fed close to ad libitum level and had free access to water. All the pigs were slaughtered at 65 kg BW. Between 30 and 65 kg BW, growth performance was significantly (p<0.001) affected by dietary treatments: average daily BW gain was 657, 530, 546 and 200 g/d for diets 1, 2, 4, and 3, respectively. Average daily DM intake was 1.8, 1.9, 2.5 and 1.4 kg/d for diets 1, 2, 4, and 3, respectively. Fat cuts (backfat+leaf fat) and backfat thickness were significantly lower on diet 3 than for other treatments (127 vs. 192, 166 g/kg of left half-carcass weight and 24.6 vs. 39.0, 35.3 mm for diet 3 vs. diets 1 and 4, and diet 2, respectively; p<0.001). The dressing weight was significantly lower on diets 2 (82.7 vs. 84.0%; p<0.001). The entire empty digestive tract (DT) weight was higher on diet 2 (73.1 vs. 66.7 g/kg empty BW). However, stomach and large intestine were more developed on diet 3: 12.8 vs. 9.3 g/100 g empty DT (p<0.001) and 26.4 vs. 23.8 g/100 g empty DT (p<0.05), respectively. In conclusion, this study suggests the CR pig has the ability to reach rather good growth and carcass performance with a well-formulated sugar cane meal and/or with a milling by-product diet refined according to its low requirements.
This experiment was conducted to investigate the effects of varying levels of metabolizable energy (ME) and crude protein (CP) on growth performance and carcass characteristics in layer-type growing male chicks. Nine hundred 1-d-old Hy-Line Brown male chicks were randomly allocated to 30 pens in a $2{\times}3$ factorial design. The experimental diets contained 2 levels of ME (2,800 kcal/kg and 2,950 kcal/kg) in combination with 3 levels of CP (17%, 18.5%, and, 20%). A significant interaction of ME and CP on feed intake was observed (p<0.05). No interaction was observed between ME and CP for 53 d BW gain or FCR, which improved linearly with dietary CP levels (p<0.05). A significant interaction or tendency was observed between ME and CP levels. The intake of ME for 1 g BW gain was linearly decreased with increasing CP levels (p<0.001). The intake of CP per bird was significantly increased in low ME (2,800 kcal/kg) treatment than that of the high ME treatment (2,950 kcal/kg) (p<0.05), and dietary CP level had more influence on CP intake for gram BW gain than level of ME. The relative weights of liver, spleen, breast meat and, leg were not influenced by the dietary treatments. Serum BUN, albumin, creatinine, and the activities of GOT and GPT were not influenced significantly by the diet treatment. In conclusion, the growth performance in layer-type male chicks was linearly increased when the level of dietary CP increased. The ME and CP did not affect the carcass characteristics and blood profiles. Therefore, the optimum levels of dietary ME and CP to improve the growth were 2,800 kcal/kg and above 18.5% in layer-type growing male chicks, respectively.
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