Negative regulation of Wee1 expression and Cdc2 phosphorylation during p53-mediated growth arrest and apoptosis

The G2 cell cycle checkpoint protects cells from potentially lethal mitotic entry after DNA damage. This checkpoint involves inhibitory phosphorylation of Cdc2 at the tyrosine-15 (Y15) position, mediated in part by the Wee1 protein kinase. Recent evidence suggests that p53 may accelerate mitotic entry after DNA damage and that the override of the G2 checkpoint may play a role in the induction of apoptosis by p53. To determine the biochemical mechanism by which p53 inactivates the G2 checkpoint, the effects of p53 activation on Wee1 expression, Cdc2-Y15 phosphorylation, and cyclin B1-associated Cdc2 kinase activity were examined. Under conditions of either growth arrest or apoptosis, p53 activation resulted in the down-regulation of Wee1 expression and dephosphorylation of Cdc2. A parallel increase in cyclin B1/Cdc2 kinase activity was observed during p53-mediated apoptosis. Negative regulation of the Wee1 expression and Cdc2 phosphorylation by p53 was also evident in thymus tissue from p53+/+ mice but not from p53-/- mice. Inactivation of the G2 checkpoint may contribute to the tumor suppressor activity of p53.     

Magnetic resonance imaging of the pelvic floor in the postpartum patient

Magnetic resonance imaging (MRI) was used to assess anatomical changes in the pelvic floor after childbirth. Six women underwent serial MRI examination within 30 hours and at 1 week, 2 weeks, 6 weeks and 6 months after delivery; 8 additional women were studied only within 30 hours of delivery. T-1 and T-2-weighted images of the pelvis in the transverse and sagittal planes with a 1.5-T MR imager were obtained. In the sagittal section we assessed the urethrovesical angle, urethral length, distance from the symphysis to the proximal and distal vagina, vaginal length, width and length of the sphincters, and the presence of sphincter defects. Axial sections were assessed for sphincter defects for the distance between the symphysis and midurethra, vagina and rectum. Only one parameter (distance between symphysis and distal vagina) changed significantly over time, without a clear trend in direction. Interobserver variation was reasonable (<15%) except for anal canal length, urethral length and distance between symphysis and anus. There were no significant correlations between birthweight and MRI parameters. There was a non-significant association (P = 0.09) between the sole combined sphincter defect and rectal injury, but not with episiotomy or parity. We concluded that it is feasible to determine multiple measurements on MR images to evaluate structures of the pelvic floor.     

Chronic administration of ORG 2766 for 6 months produces a subtle impairment in strategic learning by rats

Dysfunctions in serotonergic pathways may underlie several psychiatric disorders. The reuptake of serotonin (5-HT) from synaptic terminals is mediated by a specific transporter (5-HTT). Genetic variation in the gene coding for the 5-HTT protein might be involved in the predisposition to psychiatric disorders. A systematic screening of the whole coding sequence of the 5-HTT gene in mood disorder (MD) and obsessive-compulsive disorder (OCD) patients, as well as in healthy controls, using PCR and denaturing gradient gel electrophoresis (DGGE) revealed the presence of two mutations. The first was in intron 4, and the second was a C-->A transversion leading to an amino-acid exchange (Leu-->Met) in position 255 of the deduced protein sequence. No further occurrence of this substitution was found in an extended sample of patients and controls. Therefore, structural modifications of the 5-HTT gene do not seem to play either a major or minor role in the genetic predisposition to MD or OCD.     

Comparison of different oxygen exchange models

A functional distribution of coronary volume can be estimated from the response of arterio-venous O2 content difference (AVO2) to a flow step. However, the results depend on the assumed O2 exchange model. The previously used model consisted of a single mixed compartment with O2 exchange (reference model). The purpose of this study is to provide an estimate of the errors made in the volume estimations by not taking into account factors as flow heterogeneity, different mixing sites or Krogh-like O2 exchange. The approach is indirect: the response of the AVO2 to a flow step has been calculated with alternative O2 exchange models in which factors mentioned are incorporated. These transients are fitted with the reference model. The resulting estimated volumes are different from the volumes assumed in the alternative models. Large differences are obtained with some of the alternative models, e.g. the model with Krogh characteristics. However, these models seem unrealistic because capillary pO2 is higher than venous pO2. Only small differences in volume are obtained with the more realistic models. Therefore, these results indicate that the coronary volumes are approximated well by the estimations obtained with the reference model. These volume estimations were 9.9 and 3.8 ml 100 g-1 for the O2 exchange vessels and the distal venous volume, respectively.     

Effects of abstinence and family history for alcoholism on platelet adenylyl cyclase activity

Platelet adenylyl cyclase (AC) activity was measured in 32 alcohol-dependent subjects and 27 control subjects who were categorized as either family history-positive (FHP) or family history-negative (FHN) for alcoholism. The interview and blood sample collections were performed shortly after cessation of heavy drinking in the alcoholic group, and repeat blood samples were obtained at the end of the first and second weeks of monitored abstinence. Control subjects received the same interview and provided blood samples at the time of the interview. When subjects were not segregated for FHP or FHN status, there were no statistically significant differences in basal, cesium fluoride (CsF)-, or forskolin-stimulated mean AC activities between the controls and the alcoholics, at study entry or with 1 or 2 weeks of abstinence. On the other hand, over the 2-week course of sobriety from heavy drinking, the CsF-stimulated AC activity of FHP alcohol-dependent subjects decreased significantly (p = 0.03). FHP alcohol-dependent subjects after 2 weeks of sobriety had significantly lower mean CsF-stimulated AC activity than FHN controls (p = 0.04), whereas the FHN alcoholic subjects' CsF-stimulated AC activity did not differ significantly from FHN controls at this point in time. When all subjects were pooled and then categorized as either FHP or FHN, there was a significant difference in mean CsF-stimulated AC activity (p = 0.02) between the FHP and FHN subject groups. Genetic factors and abstinence appear to have roles in determining low platelet AC activity in alcoholic and nonalcoholic subjects. CsF-stimulated platelet AC activity, in particular, appears to act as a trait marker for a genetic vulnerability to developing alcoholism, but recent heavy drinking in male alcoholics is a factor that can mask differences between FHP and FHN subjects.     

Influence of genotype on the clinical course of the long-QT syndrome. International Long-QT Syndrome Registry Research Group

BACKGROUND: The congenital long-QT syndrome, caused by mutations in cardiac potassium-channel genes (KVLQT1 at the LQT1 locus and HERG at the LQT2 locus) and the sodium-channel gene (SCN5A at the LQT3 locus), has distinct repolarization patterns on electrocardiography, but it is not known whether the genotype influences the clinical course of the disease. METHODS: We determined the genotypes of 541 of 1378 members of 38 families enrolled in the International Long-QT Syndrome Registry: 112 had mutations at the LQT1 locus, 72 had mutations at the LQT2 locus, and 62 had mutations at the LQT3 locus. We determined the cumulative probability and lethality of cardiac events (syncope, aborted cardiac arrest, or sudden death) occurring from birth through the age of 40 years according to genotype in the 246 gene carriers and in all 1378 members of the families studied. RESULTS: The frequency of cardiac events was higher among subjects with mutations at the LQT1 locus (63 percent) or the LQT2 locus (46 percent) than among subjects with mutations at the LQT3 locus (18 percent) (P<0.001 for the comparison of all three groups). In a multivariate Cox analysis, the genotype and the QT interval corrected for heart rate were significant independent predictors of a first cardiac event. The cumulative mortality through the age of 40 among members of the three groups of families studied was similar; however, the likelihood of dying during a cardiac event was significantly higher (P<0.001) among families with mutations at the LQT3 locus (20 percent) than among those with mutations at the LQT1 locus (4 percent) or the LQT2 locus (4 percent). CONCLUSIONS: The genotype of the long-QT syndrome influences the clinical course. The risk of cardiac events is significantly higher among subjects with mutations at the LQT1 or LQT2 locus than among those with mutations at the LQT3 locus. Although cumulative mortality is similar regardless of the genotype, the percentage of cardiac events that are lethal is significantly higher in families with mutations at the LQT3 locus.     

Evaluation on non invasive diagnostic tests for the second look in epithelial ovarian cancer

OBJECTIVE: To determine the usefulness of diagnostic tests performed before a second look laparotomy in patients with epithelial ovarian cancer. STUDY DESIGN: Thirty-three patients with epithelial ovarian cancer attended at Fundación Jiménez Díaz from 1984 to 1995 were studied. All patients initially underwent cyto-reducing surgery, followed by at least six platinum-based chemotherapy cycles. Prior to second look laparotomy all patients were evaluated by computerized tomography (CT) of the pelvis and abdomen, CA-125, pelvic-abdominal echography and gynecologic examination. To evaluate sensitivity, specificity, positive predictive value and negative predictive value for each test contingency tables were used. RESULTS: Eleven out of the 33 second look patients (33%) had histologic or cytologic evidence of disease. Six out of the eleven positive second look had a positive CT prior to second look (sensitivity of 55%). CT showed lack of disease in 21 out of the 22 negative second look cases (specificity 95%). Positive and negative predictive values of the test were 86% and 81%, respectively. Nine cases out of the 28 who had a CA-125 obtained had a positive second look. Four out of these nine patients had an increased CA-125 value (sensitivity 44%, specificity 95%, positive predictive value 80% and negative predictive value 78%). Sensitivity, specificity, positive predictive value and negative predictive value of physical examination and echography were 36%, 100%, 100%, 76% and 27%, 95%, 75%, 72%, respectively. On the other hand, sensitivity, specificity, positive predictive value and negative predictive value of all tests taken together were 64%, 91%, 78% and 83%, with a rate of false-negative results of 17% and a rate of false-positive results of 22%. CONCLUSION: Pelvic-abdominal computerized tomography, CA-125, pelvic-abdominal echography and gynecologic examination can be an alternative to second look laparotomy for the diagnosis of persistence or recurrence of the disease in patients with epithelial ovarian cancer.     

Survey of the diagnosis and management of antisperm antibodies

A questionnaire was sent to all Human Fertilization and Embryology Authority-registered reproductive medicine centres throughout the UK to survey their policy for the diagnosis and management of antisperm antibodies. Forty-eight responses were received from the 74 units that use husbands' spermatozoa for treatments (65%). Most centres use at least one test to detect antibodies, although a minority perform no tests on the basis that their clinical practice would be unaltered if antibodies were present. Positive tests are classed as clinically significant at levels varying from > or = 10% to > or = 50% for direct sperm binding tests (mixed antiglobulin reaction, immunobead test), and ranging from any positive reaction to > or = 1:32 for the microtitre tests (gelatin and tray agglutination tests, microimmobilization test). Strategies for managing affected patients include no intervention, artificial insemination and intrauterine insemination (IUI) using spermatozoa prepared by various techniques, in-vitro fertilization (IVF) with or without increased insemination concentration, and intracytoplasmic sperm injection. Criteria for the latter are diverse, some centres managing all antibody-positive patients this way, while others resort to it only in severe cases or after other treatments have failed. Half of the respondents occasionally or regularly employ steroids, either alone or in conjunction with IUI or IVF. Overall, it appears that much confusion exists as to how best to manage couples presenting with antibody-related infertility.