OBJECTIVE: Human papillomavirus (HPV) is strongly implicated as a causative agent in the etiology of cervical cancer. Of its gene products, E6 and E7 oncoproteins play major roles by inactivation of cellular p53 and pRb tumor suppressor proteins, respectively. However, it has been recently suggested that p53 and/or pRb-independent functions of E6 and E7 are involved in cervical carcinogenesis. The purpose of this study is to identify novel a cellular target, p73, of E6 and to determine how E6 inactivates p73 function, METHODS: The interaction between E6 and p73 were identified by the yeast two-hybrid assay in vivo and the GST pull-down assay in vitro. The function of the interaction was determined by transient transfections using p21 promoter-CAT reporter plasmid. The molecular mechanism underlying the functional significance of the interaction was further assessed by in vivo and in vitro protein degradation assays, and gel mobility shift assays. RESULTS: Yeast two-hybrid and GST pull-down assays indicate a physical interaction between p73 and either HPV-16 or HPV-11 E6 proteins in vivo and in vitro, respectively. Transactivation domain (amino acid residues 1-49) is found to be absolutely required for this interaction. Transient co-expression of E6 significantly inhibits the p73-mediated activation of p21WAF1 promoter in a p53-defective C33A cell line. Using Ga14-p73 fusion protein, we demonstrate that E6 inhibition of p73 transactivation function is independent of sequence-specific DNA binding, which is confirmed by direct electrophoretic mobility shift assay. Moreover, E6 inhibits p73 function by interfering with the activity of the amino-terminal activation domain. The protein degradation assays in vivo and in vitro indicate that p73, unlike p53, is not susceptible to E6-dependent proteolysis. CONCLUSION: Throughout this study, we identified p73 as a novel cellular target of HPV-E6 protein and found that E6 binds p73 through the amino-terminal transactivation domain, and inhibits its transactivation function independent of the protein degradation and DNA binding. These overall results, consequently, suggest that in addition to the inactivation of p53, the functional interference of p73 by HPV-E6 may, at least in part, contribute to E6-mediated cellular transformation.
Carcinogenesis ; Cell Line ; DNA ; Electrophoretic Mobility Shift Assay ; Human papillomavirus 11 ; Human papillomavirus 16 ; Humans ; Oncogene Proteins ; Plasmids ; Proteolysis ; Transcriptional Activation ; Transfection ; Tumor Suppressor Proteins ; Two-Hybrid System Techniques ; Uterine Cervical Neoplasms ; Yeasts

No abstract available.
Hysterectomy* ; Sexual Behavior*

OBJECTIVES: AGUS often reflects an immediate cervical cancer precursor such as a HSIL mimicking an endocervical glandular lesion. In this study, we attempted to assess the clinical significance of a cytologic diagnosis of atypical glandular cells of undetermined significance (AGUS) and determine the usefulness of the human papillomavirus (HPV) DNA testing as the triage strategies in evaluating AGUS. METHODS: Between 1994 and 1998, 67,730 Papanicolaou smears were evaluated at Kangnam and Uijongbu St Mary's Hospital. There were 87 (0.13%) cases of AGUS smears during that time. Colposcopy was performed on all women, and HPV DNA testing was performed on 11 persons. RESULTS: Mean age of these patients was 45.8 years. Histologic diagnosis of AGUS were kolocytosis and CIN-I in 6 (6.9%), CIS in one, endometrial hyperplasia in 2 (2.3%), endometrial adenocarcinoma in 7 (8.0%), cervical adenocarcinoma in 14 (16.1%) and cervical squamous cell carcinoma in 2 (2.3%) cases. Endometriosis was 8.9% under 46 years old and none in over 46. CIN was 8.9% and 7.2%, respectively. Cervical adenocarcinoma was 6.7% under 46 and 19.1% over 46. Endometrial cancer was 4.4% and 11.9%, respectively. The risk of cervical cancer and endometrial cancer was high in the AGUS with Adenocarcinoma ; Carcinoma, Squamous Cell ; Colposcopy ; Diagnosis ; DNA* ; Endometrial Hyperplasia ; Endometrial Neoplasms ; Endometriosis ; Female ; Human Papillomavirus DNA Tests ; Humans ; Middle Aged ; Papanicolaou Test ; Triage ; Uterine Cervical Neoplasms

No abstract available.
Mass Screening* ; Uterine Cervical Neoplasms*

No abstract available.
Animals ; Macrophages* ; Mice*

For evaluating the reproductive performances of GTD patients, we found 115 cases of GTD patients, 77 HM and 38 GTT, who became pregnant after the completion of treatments and follow-up period. The results of this study suggest subsequent pregnancies after the completion of treatments may promise normal reproductive outcomes regardless of the chemotherapy.
Drug Therapy ; Follow-Up Studies ; Gestational Trophoblastic Disease* ; Humans ; Pregnancy

Hantavirus infection occuring during pregnancy is rarely progressing to adult respiratory distress syndrome (ARDS), which is accompanied by thrombocytopenia, leukocytosis, fever, oliguria, aggravation of bleeding, hematuria, lactacedemia. Pregnancy may have an unfavorable impact on the pathophysiologic characteristics of ARDS. The high oxygen demands of pregnancy are unable to be matched by oxygen delivery in ARDS and may predispose the patient to multiple organ failure, as well as fetal oxygen deprivation. We present the case with hantavirus pulmonary syndrome complicating pregnancy.
Fever ; Hantavirus Infections ; Hantavirus Pulmonary Syndrome* ; Hantavirus* ; Hematuria ; Hemorrhage ; Humans ; Leukocytosis ; Multiple Organ Failure ; Oliguria ; Oxygen ; Pregnancy* ; Respiratory Distress Syndrome, Adult ; Thrombocytopenia

No abstract available.
Classification* ; Uterine Cervical Neoplasms*

OBJECTIVE: Retinoic acids (RAs) and interferons (IFNs) have been implicated in the growth regulation of cervical cancer cells, which was suggested by clinical trials and in vitro experiments. However, the molecular mechanisms of growth regulation are not fully defined, The purpose of this study is to assess the effect of RA and/or IFN on human cervical carcinoma cells in vitro and to analyze their action mechanisms in HPV-positive cervical carcinoma cells by molecular biologic studies. METHODS: HPV-positive (CaSki, HeLa), HPV-negative (C33A, HT-3), and non-cervical cancer Cos-1 cell lines were treated with RA and/ar IFN. Their effects on cell growth were evaluated by the cell pmliferation assay and the following BrdU DNA incorporation assay. The molecular mechanism was further investigated by a series of immunoblottings and transient cotransfection assays, which were conducted in HeLa cells and C33A cells using the CAT reporter gene assay. To observe the down regulation of HPV E6/E7 gene expression by RA/IFN, reverse transcription-polymerase chain reaction (RT-PCR) was perforned. RESULTS: The powth of RA-treated cells was less suppressed than that of IFN-treated cells. Combined treatment of RA and IFN leads to additive effect on the growth suppression of HeLa and CaSki cells. The proliferation activity was most severely reduced in Hela cells by treatment of both all-trans-RA (AtRA) and IFN-r. Combined treatment of AtRA/IFN-r causes a great increase in the level of interferon regulatory factor-1 (IRF-1) protein in HeLa cells, whereas no induction of IRF-1 was observed in C33A cells. The CAT gene expression for IRF-1 was greatly induced by IFN-r in HeLa cells. Immunoblotting assays shows the concurrent induction of p21 CDK inhibitor and dephosphorylation of Rb protein in HeLa cells. In RT-PCR, an individual treatment of either RA or IFN reduced HPV E6/E7 mRNA levels and significantly cooperative when both RA and IFN were treated. By deaeasing E6 levels, the p53 level was increased in HeLs cells treated with RA and/or IFN. Transient cotransfection of IRF-1 and p53 as the transcription factors leads to the cooperative activation of a common p21 promoter to regulate the cell cycle. CONCLUSION: RA/IFN suppressed the growth of HPV-positive cervical cancer cells. When they were both treated, additive suppressive effects were observed in cellular proliferation as well as DNA synthesis. The growth suppressive effect is likely to be related to the increased expression of IRF-1 and p21 (antitumoral effect; p53-independent). The down regulation of HPV E6 gene suppression may account for the resultant increase of p53 levels (antiviral effect; p53-dependent). Both induced IRF-1 and p53 cooperatively augument tbe suppession of p21 CDK inhibitor, which results in dephosphorylation of pRb. Although clinical effects are likely complex and may include interactions of in vitro growth inhibitory effects with immunomodulatory and antiangiogeaetic effect, tbese results suggest the optimal clinical role for the combination of RA/IFN in the treatment of cervical canccers.
Animals ; Bromodeoxyuridine ; Cats ; Cell Cycle ; Cell Proliferation ; COS Cells ; DNA ; Down-Regulation ; Gene Expression ; Genes, Reporter ; HeLa Cells ; Humans ; Immunoblotting ; Interferon Regulatory Factor-1 ; Interferons* ; Retinoblastoma Protein ; RNA, Messenger ; Transcription Factors ; Tretinoin ; Uterine Cervical Neoplasms*

OBJECTIVES: To study the mechanism of alcohol craving is difficult because it involves both cognitive and emotional processes, which are discordamtly influenced by patients' avoidance and denial. The objective of this study is to examine the mechanism of craving by evaluating responses of emotional component of alcohol-related stimuli, explicitly and implicitly. METHODS: The subject group was composed of 19 patients with alcohol dependence, 25 heavy drinkers, and 20 social drinkers. An implicit association test (IAT) measuring differential association of 2 target concepts (alcohol versus beverage) with 2 attribute dimensions (positive versus negative) was completed. Explicit evaluation of emotional valence and arousal for alcohol-related and control stimuli was also completed. RESULTS: The patient group reported alcohol-related stimuli more negatively on both implicit and explicit evaluation. In explicit arousal evaluation, they evaluated alcohol-related stimuli more arousing than control stimuli, whereos control groups did not. While the strength of alcohol-negativity association was not related to any clinical variables, the strength of alcohol-positivity association was related to the arousal level of alcohol related stimuli, the severity of alcohol dependence, and the mean drinking amount per occasion. CONCLUSION: Alcohol craving can be regarded as negative emotion on both explicit and implicit levels in alcoholics. It seems that craving induced by alcohol-related stimuli may not be recognized by the patients but related to increased arousal or positive implicit evaluation of alcohol.
Alcoholics ; Alcoholism* ; Arousal ; Cues* ; Denial (Psychology) ; Drinking ; Humans