A Pap smear is the most important screening test for the diagnosis of cervical cancer. However, subjective judgment by the operator cannot be excluded, and replicability may greatly be reduced if uncertain specimens are examined. Examiners often experience difficulties in differentiating atrophy with inflammatory changes and ASCUS when diagnosing squamous epithelial lesions from a pap smear. Reports often vary between cytologists and pathologists, and misdiagnosis may result in delayed follow-ups and advanced diseases. Hence, auxiliary examinations are necessary when confusing results between atrophy and ASCUS are obtained. The importance of p16INK4a activation due to HPV infection, which is an important factor in the outbreak of cervical cancer, has been highlighted. Recent studies have reported that p16INK4a immunocytochemical staining and HPV high-risk type tests using liquid-based cervical specimens are effective to detect the presence of lesions of grade HSIL or higher in patients with ASC-H. However, no research exists on the utility of HPV and p16INK4a tests on the differential diagnosis of atrophy and ASCUS. This study focused on whether p16INK4a immunocytochemical staining and HPV tests can help diagnose borderline lesions between atrophy and ASCUS. The results reported that p16INK4a activation can significantly (P<0.001) differentiate atrophy from ASCUS in atrophic lesions infected with High risk-HPV. Therefore, it may be concluded that p16INK4a immunocytochemical staining is an effective auxiliary test in lesions infected with HR-HPV when atrophic lesions are difficult to differentiate by morphology. Such results are expected to help decide on adequate follow-up and treatment.
Lipids extracted from two kinds of sun-dried laver samples, wild Porphyra suborbiculata and cultured Porphyra tenera, produced at Wan-do in Korea were studied. Dried P. suborbiculata contained $0.8\%$ total lipid(TL) which consisted of $21.4\%$ neutral lipid(NL), $53.4\%$ glycolipid(GL) and $25.2\%$ phospholipid(PL), and dried P. tenera contained $1.2\%$ TL which consisted of $30.5\%$ NL, $50.3\%$ GL and $19.2\%$ PL. Among the NL of dried P. suborbiculate and P. tenera, free fatty acid ($41.4\%,\;39.0\%$), triglyceride($25.6\%,\;28.8\%$) and free sterol ($22.1\%,\;16.7\%$) were predominant. Digalactosyl diglyceride ($34.7\%,\;46.6\%$) and monogalactosyl diglyceride ($19.2\%,\;18.0\%$) were the major components among the GL. Sulfoquinovosyl digylceride ($4.2\%$) was also identified in P. tenera only. And main lipids in the PL of P. suborbiculata and P. tenera were phosphatidyl ethanolamine ($40.3\%,\;35.7\%$) and phosphatidyl choline ($28.6\%,\;30.7\%$) and followed by phosphatidyl serine($15.1\%,\;19.2\%$) and phosphatidyl inositol ($16.0\%,\;14.4\%$). The major fatty acids in the TL of the dried P. suborbiculata were 20:5 ($29.4\%$), 16:0 ($23.4\%$) and 20:4 ($13.0\%$), and those of the dried P. tenera were 20:5 ($36.7\%$), 16:0 ($16.2\%$), 16:1 ($10.7\%$) and 18:1 ($9.7\%$). The fatty acid composition of the both samples in the NL fraction were similar to the pattern in those of the TL. The abundant fatty acids in the PL of the both dried laver were 20:5, 16:0 and 18:1. In case of the GL fraction, the main fatty acids of the dried P. suborbiculata were 16:0, 20:5, 18:1 and 20:4, while those of the dried P. tenera were 20:5, 16:0 and 18:1.
The optimal culture condition of Bacillus sp. P16 was investigated for production of an extracellular endo-splitting chitosanase. The best carbon and nitrogen sources for the chitosanase production were chitosan and tryptone, respectively. The best condition for the maximum activity was at $37^{\circ}C$ in a medium containing 0.5% powdered chitosan, 1% tryptone, and 1% NaCl(at initial pH 7.0) in a rotary shaker(200 rpm). In a jar fermenter, the culture duration shortened to $6{\sim}12$ hr for maximum activity and the enzyme activity increased about 100% compared with that of flask culture.
In the present investigation, we studied the modulating effects of caffeic acid, chlorogenic acid, and (-)-epigallocatechin-3-gallate(EGCG) on the methylation status of promoter regions of cell cycle regulator, p16, in human breast cancer T-47D cells. We demonstrated that treatment of T-47D cells with caffeic acid, chlorogenic acid, or EGCG partially inhibited the methylation status of the promoter regions of p16 genes determined by methylation-specific PCR. In contrast, unmethylated p16 genes were increased with the treatment of T-47D cells with $20{\mu}M$ of caffeic acid or chlorogenic acid for 6 days. Treatment of T-47D cells with 5, 20 or $50{\mu}M$ of EGCG increased the unmethylation status of p16 gene up to 100%, and the methylation-specific bands of this gene were decreased up to 50% in a concentration-dependent manner. The finding of present study demonstrated that coffee polyphenols and EGCG have strong inhibitory effects of the cellular DNA methylation process through increased formation of S-adenosyl-homocysteine(SAH) during the catechol-O-methyltransferase (COMT)- mediated O-methylation of these dietary chemicals or an direct inhibition of the DNA methyltransferases. In conclusion, various dietary polyphenols could reverse the methylation status of p16 gene in human breast T-47D cells.
Dendritic macromolecules of the first to fourth generation were synthesized, using alkenylation and hydrosilation cycles with allylmagnesium bromide and dichloromethylsilane as building blocks and siloxane tetramer $(Me(CH_2=CH)SiO)_4$, as core molecule. By the reaction of the dichloromethylsilyl-capped generation (G4P) with p-bromophenol, p-phenylphenol and lithium phenyethynylide, dendrimers with specific functions (G4P-BP (Mw: 16,300), G4P-PP (16,121), and CAP-PA (11,764)) have been produced. Analysis of new dendrimers by NMR, UV and MALD1 mass spectrometry suggests that they are pure and unified.
Objectives: The purpose of the present study was to investigate the emotional state and personality characteristics of patient with panic disorder. Methods: Twenty patients with panic disorder, 21 patients with somatoform disorder, and 20 normal healthy controls were studied. Korean version of Beck Depression Inventory(BDI), State and Trait Anxiety Inventory(STAI) and Korean standardized edition of Cattell's 16 Personality Factors Questionnaire(16-PF) were used for assessment. Statistically, One-way ANOVA with Scheffe test were used by SPSS/PC for windows. Results: 1) Total score of BDI was higher in the panic group than the somatoform group and normal control group(p<0.001). 2) Total score of state anxiety and trait anxiety in the panic group was higher than the somatoform group and normal control group(p<0.001). 3) In 16 PF, there were no definitely abnormal range of scores. But in first-stratum source traits of 16PF, the panic group was higher than the somatoform group and normal healthy group in O-factor (p<0.01) and Q4-factor(p<0.001). In second-stratum source traits of 16PF, the panic group was higher than the somatoform group in ANX-factor(p<0.05), but lower than normal control group in TOUfactor(p<0.05). 4) There were no differences in the panic subgroup according to sex and cutoff points of BDI score 16 and STAI-T score 54. 5) According to cutoff point of STAI-S score 52, the panic subgroup above 52 was higher than the panic subgroup under 52 in G-factor(p<0.001), Q3-factor(p<0.05) and SUP-factor(p<0.001), but lower in L-factor(p<0.05). Conclusion: These results suggest that emotional state of patients with panic disorder are depressive and anxious as compared with patients with somatoform disorder and normal controls. Patients with panic disorder did not show any abnormal personality characteristics but were more guilt-prone, anxious, emotionally sensitive. We propose that the understanding of panic patients' emotional state and personality characteristics will helpful to treat and manage in patient with panic disorder.
Purpose: Methylation of gene regulatory elements plays an important role in gene inactivation without genetic alteration. Gastric cancer is one of the tumors that exhibit a high frequency of CpG island hypermethylation. The purpose of this study was to investigate the occurrence of CpG island hypermethylation in gastric carcinoma in relation to H. pylori infection, CIMP and clincopathologic variables. Materials and Methods: We investigated the promoter methylation Status of six genes (hMLH1, p16, p14, COX-2, MGMT, E-cadherin) and CIMP in 36 gastric carcinoma tissues as well as in nontumor tissues. CIMP status was investigated by examining the methylation status of MINT 1, 2, 12, 25 and 31. The methylation status of the promoter was examined by methylation-specific PCR (MSP) and H. pylori infection was examined by histological diagnosis after staining with Warthin-Starry silver. Results: Among the 36 gastric carcinoma tissues, DNA hypermethylation was detected in the following frequencies: 14 (38.9%) for p14, 13 (36.1%) for p16, 8 (22.2%) for MGMT, 10 (27.8%) for COX-2, 21 (58.3%) for E-cadherin, and 6 (16.7%) for hMLH1. The frequencies for MINT1 and MINT25 hypermethylation were significantly higher in tumor tissues than in nontumor tissues. 16 (44.4%) of the 36 gastric carcinoma tissues were positive for the CIMP CIMP-H tumors were associated with older patients and larger tumor size than CIMP-L tumors. We found a significant association between the presence of the CIMP and hypermethylation of p16. Hypermethylation of p16 and MINT2 were significantly different when compared by age. MINT1 gene methylation was significantly associated with H. pylori infection (P=0.004). Conclusion: Our results suggest that aberrant hypermethylation of multiple tumor related genes (hMLH1, p16, p14, COX-2, MGMT, E-cadherin, MINT1, 2, 12, 25, 31) occurs frequently in gastric carcinoma tissues. The hypermethylation of MINT1 was significantly higher in the tumor tissues and was associated with H. pylori infection.
BACKGROUND/OBJECTIVES: Previous studies have indicated that when compared to young mice, old mice have lower global DNA methylation and higher p16 promoter methylation in colonic mucosa, which is a common finding in colon cancer. It is also known that a Western-style diet (WSD) high in fat and calories, and low in calcium, vitamin D, fiber, methionine and choline (based on the AIN 76A diet) is tumorigenic in colons of mice. Because DNA methylation is modifiable by diet, we investigate whether a WSD disrupts DNA methylation patterns, creating a tumorigenic environment. SUBJECTVIES/METHODS: We investigated the effects of a WSD and aging on global and p16 promoter DNA methylation in the colon. Two month old male C57BL/6 mice were fed either a WSD or a control diet (AIN76A) for 6, 12 or 17 months. Global DNA methylation, p16 promoter methylation and p16 expression were determined by LC/MS, methyl-specific PCR and real time RT-PCR, respectively. RESULTS: The WSD group demonstrated significantly decreased global DNA methylation compared with the control at 17 months (4.05 vs 4.31%, P = 0.019). While both diets did not change global DNA methylation over time, mice fed the WSD had lower global methylation relative to controls when comparing all animals (4.13 vs 4.30%, P = 0.0005). There was an increase in p16 promoter methylation from 6 to 17 months in both diet groups (P < 0.05) but no differences were observed between diet groups. Expression of p16 increased with age in both control and WSD groups. CONCLUSIONS: In this model a WSD reduces global DNA methylation, whereas aging itself has no affect. Although the epigenetic effect of aging was not strong enough to alter global DNA methylation, changes in promoter-specific methylation and gene expression occurred with aging regardless of diet, demonstrating the complexity of epigenetic patterns.
Background: Cancer of the gallbladder is a relatively rare neoplasm with a poor prognosis. The exact mechanisms of its genesis are not known and very little information is available on molecular events leading to labeling this as an orphan cancer. Materials and Methods: In this prospective case control study we evaluated the expression of p16, pRb and cyclin D1 by immunohistochemistry to study the G1-S cell-cycle check point and its possible role in gallbladder carcinogenesis. A total of 25 patients with gallbladder carcinoma (group I), 25 with cholelithiasis (group II) and 10 normal controls. were enrolled Results: Cyclin D1 expression was seen in 10 (40%) patients each with carcinoma and cholelithiasis while only in 2 (20%) of the normal gallbladders but differences were not statistically significant (p value=0.488). p16 was expressed in 12% patients of carcinoma of the gallbladder and 28% of cholelithiasis, however this difference was not statistically significant (p value=0.095). Retinoblastoma protein was found to be expressed in 50% of normal gallbladders and 6 (24%) of carcinoma and 8 (32%) of gallstones. The present study failed to demonstrate any conclusive role of cyclin D1/RB/ p16 pathway in carcinoma of the gallbladder. Conclusions: The positive relation observed between tumor metastasis and cyclinD1 expression and p16 with nodal metastasis suggested that higher cyclin D1/p16 expression may act as a predictive biomarker for aggressive behavior of gallbladder malignancies.
This study was conducted to investigate the optimal CP level in growing goat kids. Forty male goats were divided into four treatment groups fed diets containing CP 12, 14, 16, and 18% of concentrate feed, and rice straw, respectively. Results are summarized as follows. Average daily gain’s(ADG) of groups fed diets with CP 14% and CP 18% were 84.0 and 83.0g/d each, which were higher than group fed diets with CP 12% grown at 69.2g of ADG(P<0.05). Daily feed intakes of concentrate and rice straw were 590g and 45g each and there was no difference found between treatments. The amounts of feed required for body weight gain(g) were similar in the range of 7.0-7.3 for groups fed diets with CP 14%, 16% or 18%. But the group fed diets with CP 12% required somewhat more feed for gain at 8.8g. Dressing percentage of groups fed diet with CP 12% was 61.7%, which was higher than groups fed diets with CP 14% or CP 18%(P<0.05) but similar to that fed diet with CP 16%. Meat percentage from goats fed diet with CP 16% was 51.7%, which were higher than goats fed diets with CP 12% or CP 14%(P<0.05). Fat deposition of CP 12% group was higher than the other groups(P<0.05). Percentage of bone weight averaged 17.0% without any difference among treatment groups. Shear force and cooking loss, which are physical properties of goat meats, were lower in CP 16% and CP 14% groups than the other two groups. Water holding capacity of goats meats from CP 16% was higher than those from CP 18% group(P<0.05). Results from panel test showed that juiciness of meats from CP 16% group was higher than those from CP 12% group(P<0.05). Meats from CP 16% also was tested to be more tender than meats from the other groups. Results from this work suggest that the optimum crude protein level in growing goat's concentrate is 14-16% and that increase above this level seems not to improve meat production.
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