Evaluating the expression of urokinase and tissue leukocyte being in benign and malignant breast disease

Introduction: Our objectives is to show that the expression of uPA leukocyte could be considered, in the future, as a marker of the expression of uPA in the malignant tissue and therefore a potential indicator of prognosis. Methods: We examined the expression of uPa in leukocytes and tissues of three groups of women: with breast cancer; with benign breast lesion and healthy women (control group). We used RT Real Time PCR assay. The expression of urokinase is significantly higher in malignant breast lumps compared to benign lesions. However, in women with carcinoma of the breast, malignant tissue expresses higher amounts of uPA than the healthy counterpart. There are no statistically significant differences in the expression of uPA, between tissues taken from women with benign lesions. The lymphocytes taken from healthy volunteers show a level of expression of uPA significantly lower than the other tested samples Lymphocytes extracted from cancer patients express higher amounts of uPA compared to lymphocytes belonging to women with benign breast lesions. The expression of uPA was compared with the clinical and biological parameters commonly used in clinical practice for the definition of the prognosis. The only exception found, concerns those tumors characterized by the simultaneous negativity for estrogen receptors, progesterone and HER2 (state of triple negative), in which the expression of uPA is very high. Results and conclusions: Our data show that uPA expressed by leukocytes of each individual patient is the mirror image of the one expressed by malignant nodular uPA.Introduction: Our objectives is to show that the expression of uPA leukocyte could be considered, in the future, as a marker of the expression of uPA in the malignant tissue and therefore a potential indicator of prognosis. Methods: We examined the expression of uPa in leukocytes and tissues of three groups of women: with breast cancer; with benign breast lesion and healthy women (control group). We used RT Real Time PCR assay. The expression of urokinase is significantly higher in malignant breast lumps compared to benign lesions. However, in women with carcinoma of the breast, malignant tissue expresses higher amounts of uPA than the healthy counterpart. There are no statistically significant differences in the expression of uPA, between tissues taken from women with benign lesions. The lymphocytes taken from healthy volunteers show a level of expression of uPA significantly lower than the other tested samples Lymphocytes extracted from cancer patients express higher amounts of uPA compared to lymphocytes belonging to women with benign breast lesions. The expression of uPA was compared with the clinical and biological parameters commonly used in clinical practice for the definition of the prognosis. The only exception found, concerns those tumors characterized by the simultaneous negativity for estrogen receptors, progesterone and HER2 (state of triple negative), in which the expression of uPA is very high. Results and conclusions: Our data show that uPA expressed by leukocytes of each individual patient is the mirror image of the one expressed by malignant nodular uPA

Italian randomized trial among womenwith histerectomy: tamoxifen and hormone-dependent breast cancer in hight-risk women

Abstract Tamoxifen improves outcome in women with breast cancer and reduces the incidence of estrogen receptor-positive (ER+) breast tumors in prevention trials. Tamoxifen use is associated with an increased risk of potentially serious adverse events, principally endometrial cancer and venous thromboembolic events and, therefore, detailed knowledge of the effects of tamoxifen is important. With more cases of breast cancer being found as the follow-up time increases, it is now possible to perform more detailed analysis of the Italian Randomized Trial of Tamoxifen. Women with hysterectomy (N = 5408) were randomly assigned to receive 20 mg tamoxifen per day (N = 2700) or placebo (N = 2708). After a median of 81.2 months of follow-up, 79 case subjects (34 in the tamoxifen arm and 45 in the placebo arm) were diagnosed with breast cancer. We were able to identify a group of women at increased risk of ER+ breast cancers (high-risk group) on the basis of baseline as well as reproductive and hormonal characteristics (height, age at menarche, parity, age at first birth, and oophorectomy). Tamoxifen administered to women in the high-risk group showed statistically significantly reduced incidence of breast cancer (tamoxifen, 3 and placebo, 15; P =.003), but no such effect was seen in the low-risk group (tamoxifen, 31 and placebo, 30; P =.89). The positive effect of tamoxifen on breast cancer among high-risk women is most marked for ER+ tumors (tamoxifen, 1 and placebo, 11; P =.002). Chemoprevention of breast cancer with tamoxifen appears to be effective in women at high risk of ER+ tumors but not among women at low risk, who may well be protected naturally by late age at menarche or early first pregnancy, or artificially by removal of the ovaries. Tamoxifen could be offered as a preventive agent to women identified at high-risk of breast cancer because of hormone-related risk factors. Such a strategy would greatly reduce the numbers of women who would need to take tamoxifen to obtain the same absolute reduction in breast cancer. These findings are exploratory and need to be confirmed in other randomized trials

FLAT FLAME BURNER ANALYSES

Velocity and temperature fields in the sintered material surrounding an embedded cooling coil in a Kaskan type flat flame burner are predicted. An approximate two-dimensional velocity field is obtained in closed form using a potential flow analogy. The velocity profile above the burner surface will be flat if the distance from the burner surface to the plane of the coil is sufficiently large that the coil wake is ameliorated. Quantitative results for this minimum distance are presented. Several approximations to the temperature field are discussed. Detailed temperature profiles are given for the simplest - one-dimensional with uniform velocity - and the most complex - two-dimensional with potential flow velocity field. The influence of the burner and cooling geometries, fuel type and stoichiometry, flow rate, ambient and cooling water temperatures and sintered material properties, are all described by a small set of dimensionless parameters. Optimization of burner performance in terms of these parameters is discussed. It is hoped that the results will be useful in the design of burners for flame structure studies

Dual Effect of Methylprednsolone Pulses on Apoptosis of Peripheral Leukocytes in Patients with Renal Diseases

It is well known that change in apoptosis may modulate the natural story of illness, and that many drugs may act through modulation of apoptosis, but the role of steroids in acting through apoptosis in different settings, including renal diseases, has still to be elucidated. We studied the in vivo effects of steroids by oral assumption (10 to 25 mg/deltacortene) or by intravenous pulses (300 to 1000 mg/dose) on apoptosis and cellular subsets of peripheral lymphocytes, by evaluating DNA-fragmentation and lymphocyte subsets in 79 subjects: 22 controls and 57 patients with various renal diseases (25 Lupus-GN, 19 membranous-GN (MGN), 6 rapidly progressive-GN (RPGN), 2 acute interstitial nephritis (AIN), 5 on chronic dialysis. Baseline apoptosis was present in 1/22 (4.5%) of controls, 3/25 (12%) SLE, 2/6 (33.3%) RPGN and 10/19 (52.6%) MGN. A significant decrease in CD3+CD8+ cell count and a significant increase of the CD3+CD4/CD3+CD8+ ratio were found in apoptosis-positive subjects. DNA fragmentation did not change after oral steroids, paralleling a 22 to 32% decrease in total lymphocytes. Following intravenous methylprednisolone pulses, a deeper drop of all lymphocyte subsets was observed, while DNA fragmentation turned from present to absent in 2 MGN, but not in 2 RPGN, and from absent to present in 1 ARF and 1 SLE, independently of the dosage. We demonstrated that the presence of apoptosis in renal diseases is associated with decreased CD3+CD8+ cell count. Furthermore, steroid intravenous pulses, besides inducing a profound decrease in lymphocyte subsets, do exert a dual effect on baseline leukocyte apoptosis, eventually leading to a reversal of baseline patterns, either turning from negative to positive or from positive to negative. Oral steroid therapy did not influence baseline apoptosis

Prevention of Breast cancer with tamoxifen:preliminary findings from the italian randomised trial among hysterectomised women

Prevention of breast cancer with tamoxifen: preliminary findings from the Italian randomised trial among hysterectomised women. Italian Tamoxifen Prevention Study. Veronesi U, Maisonneuve P, Costa A, Sacchini V, Maltoni C, Robertson C, Rotmensz N, Boyle P. SourceEuropean Institute of Oncology, Milan, Italy. Abstract BACKGROUND: Tamoxifen is a candidate chemopreventive agent in breast cancer, although the drug may be associated with the development of endometrial cancer. Therefore we did a trial in hysterectomised women of tamoxifen as a chemopreventive. METHODS: In October, 1992, we started a double-blind placebo-controlled, randomised trial of tamoxifen in women (mainly in Italy) who did not have breast cancer and who had had a hysterectomy. Women were randomised to receive tamoxifen 20 mg per day or placebo, both orally for 5 years. The original plan was to follow the intervention phase by 5 years' follow-up. In June, 1997, the trialists and the data-monitoring committee decided to end recruitment primarily because of the number of women dropping out of the study. Recruitment ended on July 11, 1997, and the study will continue as planned. The primary endpoints are the occurrence of and deaths from breast cancer. This preliminary interim analysis is based on intention-to-treat. FINDINGS: 5408 women were randomised; participating women have a median follow-up of 46 months for major endpoints. 41 cases of breast cancer occurred so far; there have been no deaths from breast cancer. There is no difference in breast-cancer frequency between the placebo (22 cases) and tamoxifen (19) arms. There is a statistically significant reduction of breast cancer among women receiving tamoxifen who also used hormone-replacement therapy during the trial: among 390 women on such therapy and allocated to placebo, we found eight cases of breast cancer compared with one case among 362 women allocated to tamoxifen. Compared with the placebo group, there was a significantly increased risk of vascular events and hypertriglyceridaemia among women on tamoxifen. INTERPRETATION: Although this preliminary analysis has low power, in this cohort of women at low-to-normal risk of breast cancer, the postulated protective effects of tamoxifen are not yet apparent. Women using hormone-replacement therapy appear to have benefited from use of tamoxifen. There were no deaths from breast cancer recorded in women in the study. It is essential to continue follow-up to quantify the long-term risks and benefits of tamoxifen therapy

HPV sensitizes OPSCC cells to cisplatin-induced apoptosis by inhibiting autophagy through E7-mediated degradation of AMBRA1

Oropharyngeal squamous cell carcinoma (OPSCC) is an increasing world health problem with a more favorable prognosis for patients with human papillomavirus (HPV)-positive tumors compared to those with HPV-negative OPSCC. How HPV confers a less aggressive phenotype, however, remains undefined. We demonstrated that HPV-positive OPSCC cells display reduced macroautophagy/autophagy activity, mediated by the ability of HPV-E7 to interact with AMBRA1, to compete with its binding to BECN1 and to trigger its calpain-dependent degradation. Moreover, we have shown that AMBRA1 downregulation and pharmacological inhibition of autophagy sensitized HPV-negative OPSCC cells to the cytotoxic effects of cisplatin. Importantly, semi-quantitative immunohistochemical analysis in primary OPSCCs confirmed that AMBRA1 expression is reduced in HPV-positive compared to HPV-negative tumors. Collectively, these data identify AMBRA1 as a key target of HPV to impair autophagy and propose the targeting of autophagy as a viable therapeutic strategy to improve treatment response of HPV-negative OPSCC. Abbreviations: AMBRA1: autophagy and beclin 1 regulator 1; CDDP: cisplatin (CDDP); FFPE: formalin-fixed paraffin-embedded (FFPE); HNC: head and neck cancers (HNC); HPV: human papillomavirus (HPV); hrHPV: high risk human papillomavirus (hrHPV); OCSCC: oral cavity squamous carcinomas (OCSSC); OPSCC: oropharyngeal squamous cell carcinoma (OPSCC); OS: overall survival (OS); qPCR: quantitative polymerase chain reaction; RB1: RB transcriptional corepressor 1; ROC: receiver operating characteristic curve (ROC)