PURPOSE: This study was performed to examine any histopathological changes occurring in the growth plate when the rats were subjected to be deprived of normal weight bearing using the hindlimb suspension model, and to search for any countermeasures for improving and/or recovering the chondrocyte activities within the growth plate. MATERIALS AND METHODS: Sixty male Sprague-Dawley rats, aged 6 weeks, were divided into 10 groups each: Group I-control to unloading; Group II-unloading 3 weeks only; Group III-unloading+application of heat shock; Group IV-unloading+application of antioxidant; Group V-unloading+application of heat shock and antioxidant; Group VI-control to reloading; Group VII-reloading 1 week only; Group VIII-reloading+application of heat shock; Group IX-reloading+application of antioxidant; Group X-reloading+application of heat shock and antioxidant. The animals were double labeled with 5-Bromo-2'-deoxydiuridin (BrdU) and BrdU immunohistochemistry was performed for the cellular kinetic analysis. Transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) assay was done for the investigation of apoptotic changes in the growth plate, and the positive cells were counted in each zones of the growth plate in both TUNEL and BrdU immunohistochemistry. Heat shock protein (HSP), indian hedgehog (Ihh), and vascular endothelial growth factor (VEGF) were immunolocalized to assess the chondrocytic activities in terms of production of extracellular matrix protein. RESULTS: Non-weight bearing induced a reduction of height of the growth plate, reduced cellular proliferation of chondrocytes, reduced expression of Ihh and VEGF, and altered expression of heat shock protein. When heat shock and/or antioxidant were applied to the unloaded and reloaded rats, only rats in the group of application of both heat shock and antioxidant showed normal cellular activities in terms of cellular proliferation and the production of extracellular matrix protein. CONCLUSION: The present results suggest that application of heat shock and antioxidant would be a countermeasure for the restoration of chondrocytic activities when the normal weight-bearing is deprived of.
Aged ; Animals ; Bromodeoxyuridine ; Cell Proliferation ; Chondrocytes ; Deoxyuridine ; Extracellular Matrix ; Growth Plate ; Heat-Shock Proteins ; Hedgehogs ; Hindlimb Suspension ; Hot Temperature ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Rats ; Rats, Sprague-Dawley ; Shock ; Ursidae ; Vascular Endothelial Growth Factor A ; Weight-Bearing

OBJECTIVE: To investigate the change of placental apoptosis and the expression of their mediator in preeclampsia women. METHODS: Placental tissues from 10 cases of preeclampsia and 15 cases of normal pregnancy were analyzed using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling(TUNEL) staining. Expressions of bcl-2, bax, caspase-3 was also assessed using immunohistochemistry. RESULTS: In TUNEL staining, the number of apoptotic nuclei were significantly increased in the trophoblast of preeclampsia than normal pregnancy. Bcl-2 was mainly expressed in syncytiotrophoblast and bax was mainly expressed in cytotrophoblast. Bcl-2 expression was decreased and bax expression was increased in the preeclampsia than normal, but the difference was not significant. Caspase-3 was mainly expressed in the cytotrophoblast and expression was significantly increased in the preeclampsia than normal pregnancy(p<0.05). CONCLUSION: Placental apoptosis, especially accompanied with increased expression of caspase-3 in cytotrophoblast, might be related with in the pathogenesis of preeclampsia.
Apoptosis* ; Caspase 3 ; Deoxyuridine ; Female ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Placenta ; Pre-Eclampsia* ; Pregnancy ; Trophoblasts

OBJECTIVE: The purpose of this study is to determine the time evolution and distribution of cerebral apoptosis using the middle cerebral artery occlusion model in rats. METHODS: A total of twenty four male rats - with 2, 3, 4, 6, 8, 12, 24 and 48 hours of middle cerebral artery occlusion respectively - were studied. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick-end labeling(TUNEL) method was used for the observation of the apoptotic cells. The apoptotic ratio was calculated and the distribution of apoptosis was inspected in the pyriform cortex, basal ganglia and middle cerebral artery territory cortex. The rats were divided into three groups(Group I: 2~4 hours of occlusion, Group II: 6~12 hours of occlusion, Group III: 24~48 hours of occlusion). RESULTS: In this study, the proportion of apoptosis increased with the duration of middle cerebral artery occlusion and reached a maximum after about 12 hours of middle cerebral artery occlusion. The mean values of the apoptotic ratio were 30.7+/-11.3% in group I, 60.8+/-2.6% in group II and 48.7+/-0.7% in group III. The distribution of apoptosis differed in the pyriform cortex, basal ganglia and middle cerebral artery territory cortex according to the duration of time of the middle cerebral artery occlusion. CONCLUSION: In the middle cerebral artery occlusion model of the rats, apoptosis is found to increase according to the occlusion time, reaching a peak after 6 hours, and the distribution of apoptosis changed from the pyriform cortex to the basal ganglia and middle cerebral artery territory cortex.
Animals ; Apoptosis* ; Basal Ganglia ; Deoxyuridine ; Humans ; Infarction, Middle Cerebral Artery* ; Ischemia ; Male ; Middle Cerebral Artery* ; Rats*

Here, we describe a uracil-DNA glycosylase (UNG)-treated reverse transcription loop-mediated isothermal amplification (uRT-LAMP) for the visual detection of all subtypes of avian influenza A virus (AIV). The uRT-LAMP assay can prevent unwanted amplification by carryover contamination of the previously amplified DNA, although the detection limit of the uRT-LAMP assay is 10-fold lower than that of the RT-LAMP without a UNG treatment. To the best of our knowledge, this is the first successful application of deoxyuridine triphosphate/UNG strategy in RT-LAMP for AIV detection, and the assay can be applied for the rapid, and reliable diagnosis of AIVs, even in contaminated samples.
Animals ; Deoxyuridine ; Diagnosis ; DNA ; Influenza in Birds* ; Limit of Detection ; Reverse Transcription* ; Uracil-DNA Glycosidase

OBJECTIVE: The purpose of this study was to investigate the degree of apoptosis in placentas with pregnancy complicated by fetal growth restriction or preeclampsia as compared with normal term pregnancy placentas and to evaluate the expression of proapoptotic proteins Bax, p53 and antiapoptotic protein Bcl-2 in their placentas. METHODS: Placentas were obtained from 40 normal term pregnancies and from 30 pregnancies complicated by fetal growth restriction or preeclampsia admitted for delivery to the department of Obstetrics and Gynecology of Inha university hospital from January 1 to November 30, 2003. Placental sections were examined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) staining, indicative of apoptosis. The expressions of Bcl-2, Bax, p53 in their placentas were analyzed by immunohistochemical staining. RESULTS: Increased apoptosis was found in the trophoblast layer of villi from pregnancies complicated by fetal growth restriction or preeclampsia (n=30) compared to normal pregnancies (n=40) (0.93 +/- 0.54: 1.5 +/- 1.2) (p=0.014, t-test). There was no statistical significance between the two groups in expressions of Bcl-2, Bax, p53 (p=0.073, p=0.424, p=0.208, chi-square test). CONCLUSION: These results suggest that placental apoptosis may play a role in the pathogenesis of fetal growth restriction or preeclampsia. But placental apoptosis does not seem to be mediated by Bcl-2, Bax, p53 in trophoblasts.
Apoptosis* ; Deoxyuridine ; Fetal Development* ; Gynecology ; Obstetrics ; Placenta ; Pre-Eclampsia* ; Pregnancy* ; Trophoblasts

Apoptosis, including the programmed cell death, is important event in normal cell turnover and maintenance of adult tissues. Apoptosis exerts a homeostatic function in relation to tissues dynamics, as the steady state of continuously renewing tissues achieved by a balance between cell replication and cell death. This study was undertaken to investigate the association between apoptosis and development of the cervical neoplasia. Archival cervical samples from normal epithelium (n 10), low-grade squamous intraepithelial lesions (LSIL, n = 10), high-grade squamous intraepithelial lesions (HSIL, n 10), microinvasive squamous cell carcinomas (n 10), and invasive squamous cell carcinomas (n = 10) were evaluated for apoptosis. We used in situ end-labeling of DNA strand breaks by terminal deoxynucleotidyltransferase incorporation of biotinylated deoxyuridine to 3-OH ends of DNA, identified by nickel-avidine-peroxidase. The apoptotic index (sum of apoptotic bodies divided by the total nuclei times 100) significantly decreased (P<0.05) as the degree of neoplasia increased: 3.1 + 0.9 % in normal epithelium, 5.5 +/- 1.4 % in LSIL, 1.6 +/- 0.4 % in HSIL, 1.9 +/- 0.5 % in microinvasive carcinomas, and 0.6 +/- 0.3 % in invasive carcinomas. Compared to normal epithelium, the total cell number per 200x field increased significantly (P<0,05): 379 +/- 47 in normal epithelium, 462 +/- 228 in LSIL, 670+/-293 in HSIL, 1035 +/- 254 in microinvasive carcinomas, and 1389 +/- 247 in invasive carcinomas. Consequently, these results suggest that progession of cervical carcinogenesis is associated with a decrease in apoptotic index and an increase in the number of the total cell.
Adult ; Apoptosis* ; Carcinogenesis ; Carcinoma, Squamous Cell ; Cell Count ; Cell Death ; Deoxyuridine ; DNA ; DNA Nucleotidylexotransferase ; Epithelium ; Humans

BACKGROUND: Survivin, a novel antiapoptotic gene has been linked with tumor development and progression in various human carcinomas including gastric carcinomas. The aim of this study was to evaluate the expression of survivin in gastric carcinoma and its correlation with tumor cell proliferation and apoptosis. METHODS: Expression of survivin was evaluated immunohistochemically in 84 surgically resected gastric carcinomas. Tumor cell apoptosis was evaluated with terminal deoxynucleotidyl transferase (TdT) mediated deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL), and Ki-67 immunostaining was used for evaluation of tumor cell proliferation. RESULTS: Expression of survivin was noted in 53.6% of the gastric carcinomas, and was significantly associated with depth of invasion, status of lymph node metastasis or tumor stage (p=0.022, 0.034, 0.040, respectively). There was an inverse correlation between survivin expression and apoptotic index (p=0.015). But there was no significant correlation between survivin expression and Ki-67 labeling index (p=0.430). CONCLUSIONS: These results suggest that survivin expression may contribute to tumor development and progression by inhibiting apoptosis in human gastric carcinoma.
Apoptosis ; Cell Proliferation ; Deoxyuracil Nucleotides ; Deoxyuridine ; DNA Nucleotidylexotransferase ; Humans ; Lymph Nodes ; Neoplasm Metastasis ; Stomach Neoplasms

BACKGROUND: Thymidylate synthase (TS) catalyzes the methylation of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP), and this is an essential step in DNA biosynthesis. The present investigation was designed to determine the expression of TS in the patients with non-small cell lung cancer (NSCLC) and to assess the possible associations between the TS status and the p53 or proliferative index (PI). METHODS: The archival tumor tissues from 56 previously untreated NSCLC patients were examined by immunohistochemistry for TS, p53 and Ki-67. RESULTS: Forty-one men and 15 women (age range: 35 to 79 years, mean age: 62 years) were included in this study. The TS expression was high in 40 patients (71.4%) and low in 16 patients (28.6%). The aberrant expression of p53 was detected in 35 patients (62.5%). The mean PI for all the patients was 31.4+/-12.1. The TS-high tumors tended to be more poorly differentiated (p=0.069). The TS expression by a semiquantitative fourscale grading system was significantly correlated with the PIs (p=0.003). No correlation was established between the TS expression and the p53 status (p=0.806) or survival (p=0.951). CONCLUSIONS: TS was not confirmed to be a useful marker for determining the prognosis of NSCLC patients. However, our data suggest that the tumor cells with higher TS expression have a higher proliferative activity.
Carcinoma, Non-Small-Cell Lung* ; Deoxyuridine ; DNA ; Female ; Humans ; Immunohistochemistry ; Lung Neoplasms ; Male ; Methylation ; Prognosis ; Thymidine ; Thymidylate Synthase*

OBJECTIVE: Our purpose was to investigate the incidence of placental apoptosis in pregnancies complicated by preeclampsia. METHODS: Placental samples were obtained from 15 uncomplicated third-trimester pregnancies and from 17 pregnancies complicated by preeclampsia. TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling) staining and electron microscopy were performed on all placental samples and identified apoptosis within the cells of the placenta. TUNEL positive stained cells were counted at each photograph(five sections were photoragphed at 1,500 magnifications for each sample). We focused on only the syncytiotrophoblast nuclei of the placenta. The number of apoptotic syncytiotrophoblast nuclei identified was expressed as a percentage of total number of syncytiotrophoblast nuclei counted. RESULTS: Quantification of apoptosis (mean+/-SD) was as follows: normal third trimester (n=15) 0.57+/- 0.47% of cells and preeclampsia third trimester (n=17) 1.41+/-0.67% of cells. The incidence of apoptosis was significantly increased in placentas from pregnancies with preeclampsia compared with normal placentas (p< 0.01). CONCLUSION: These results suggest that placental apoptosis may play a role in the pathophysiologic mechanisms of preeclampsia.
Apoptosis* ; Deoxyuridine ; Female ; Humans ; In Situ Nick-End Labeling ; Incidence ; Microscopy, Electron ; Placenta ; Pre-Eclampsia* ; Pregnancy ; Pregnancy Trimester, Third ; Trophoblasts

BACKGROUND: Cancer stem cells have been investigated as new targets for colorectal cancer (CRC) treatment. We recently reported that CD133+ colon cancer cells showed chemoresistance to 5-fluorouracil through increased survivin expression and proposed the survivin inhibitor YM155 as an effective therapy for colon cancer in an in vitro study. Here, we investigate the relationship between survivin and CD133 expression in surgically resected CRC to identify whether the results obtained in our in vitro study are applicable to clinical samples. METHODS: We performed immunohistochemical staining for survivin and CD133 in surgically resected tissue from 187 stage II or III CRC patients. We also comparatively analyzed apoptosis according to survivin and CD133 expression using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling. RESULTS: The results of the Mantel-Haenszel test established a linear association between nuclear survivin and CD133 expression (p = .018), although neither had prognostic significance, according to immunohistochemical expression level. No correlation was found between survivin expression and the following pathological parameters: invasion depth, lymph node metastasis, or histologic differentiation (p > .05). The mean apoptotic index in survivin+ and CD133+ tumors was higher than that in negative tumors: 5.116 ± 4.894 in survivin+ versus 4.103 ± 3.691 in survivin– (p = .044); 5.165 ± 4.961 in CD133+ versus 4.231 ± 3.812 in CD133– (p = .034). CONCLUSIONS: As observed in our in vitro study, survivin expression is significantly related to CD133 expression. Survivin may be considered as a new therapeutic target for chemoresistant CRC.
Apoptosis ; Colonic Neoplasms ; Colorectal Neoplasms* ; Deoxyuridine ; Fluorouracil ; Humans ; In Vitro Techniques ; Lymph Nodes ; Neoplasm Metastasis ; Neoplastic Stem Cells