Recent climate tendencies on an East Antarctic ice shelf inferred from a shallow firn core network.

Nearly three decades of stable isotope ratios and surface mass balance (SMB) data from eight shallow firn cores retrieved at Fimbul Ice Shelf, East Antarctica, in the Austral summers 2009-2011 have been investigated. An additional longer core drilled in 2000/2001 extends the series back to the early eighteenth century. Isotope ratios and SMB from the stacked record of all cores were also related to instrumental temperature data from Neumayer Station on Ekström Ice Shelf. Since the second half of the twentieth century, the SMB shows a statistically significant negative trend, whereas the δ18O of the cores shows a significant positive trend. No trend is found in air temperature at the nearest suitable weather station, Neumayer (available since 1981). This does not correspond to the statistically significant positive trend in Southern Annular Mode (SAM) index, which is usually associated with a cooling of East Antarctica. SAM index and SMB are negatively correlated, which might be explained by a decrease in meridional exchange of energy and moisture leading to lower precipitation amounts. Future monitoring of climate change on the sensitive Antarctic ice shelves is necessary to assess its consequences for sea level change. KEY POINTS: Mass balance and stable oxygen isotope ratios from shallow firn coresDecreasing trend in surface mass balance, no trend in stable isotopesNegative correlation between SAM and SMB.

Chronology of Pu isotopes and 236U in an Arctic ice core.

In the present work, state of the art isotopic fingerprinting techniques are applied to an Arctic ice core in order to quantify deposition of U and Pu, and to identify possible tropospheric transport of debris from former Soviet Union test sites Semipalatinsk (Central Asia) and Novaya Zemlya (Arctic Ocean). An ice core chronology of (236)U, (239)Pu, and (240)Pu concentrations, and atom ratios, measured by accelerator mass spectrometry in a 28.6m deep ice core from the Austfonna glacier at Nordaustlandet, Svalbard is presented. The ice core chronology corresponds to the period 1949 to 1999. The main sources of Pu and (236)U contamination in the Arctic were the atmospheric nuclear detonations in the period 1945 to 1980, as global fallout, and tropospheric fallout from the former Soviet Union test sites Novaya Zemlya and Semipalatinsk. Activity concentrations of (239+240)Pu ranged from 0.008 to 0.254 mBq cm(-2) and (236)U from 0.0039 to 0.053 µBq cm(-2). Concentrations varied in concordance with (137)Cs concentrations in the same ice core. In contrast to previous published results, the concentrations of Pu and (236)U were found to be higher at depths corresponding to the pre-moratorium period (1949 to 1959) than to the post-moratorium period (1961 and 1962). The (240)Pu/(239)Pu ratio ranged from 0.15 to 0.19, and (236)U/(239)Pu ranged from 0.18 to 1.4. The Pu atom ratios ranged within the limits of global fallout in the most intensive period of nuclear atmospheric testing (1952 to 1962). To the best knowledge of the authors the present work is the first publication on biogeochemical cycles with respect to (236)U concentrations and (236)U/(239)Pu atom ratios in the Arctic and in ice cores.

Effects of long-term HRT and tamoxifen on the expression of progesterone receptors A and B in breast tissue from surgically postmenopausal cynomolgus macaques.

Estrogen is a well-known mitogen in breast epithelium but the role of progesterone is complex and incompletely understood. In contrast to what is seen in the endometrium, combined estrogen/progestogen treatment for postmenopausal replacement (HRT) may carry a risk for breast cancer beyond that of estrogen alone. The ratio of the two progesterone receptor (PR) isoforms, PRA/PRB may define the response to progesterone in reproductive tissues. In a primate model for long-term HRT, surgically, postmenopausal cynomolgus macaques were treated for 35 months with conjugated equine estrogens (CEE), medroxyprogesterone acetate (MPA), CEE + MPA and tamoxifen (n = 5 in all groups). The immunohistochemical expression of PRA, PRB and the androgen receptor (AR) in breast tissue was quantified by image analysis. Over all, the total PR immunostaining in glandular epithelium was more abundant during CEE (mean 12%) and tamoxifen ( 1%) treatment as compared to CEE/MPA (5%), MPA (4%) and untreated controls (6%). Differences in PRB expression were observed between treatment groups (p < 0.05). In the CEE group levels of PRA were unchanged while there was a decline in the CEE/MPA group. The mean PRA/PRB ratio in the CEE group was 2.7 and in the CEE/MPA group 0.2. Treatment with tamoxifen had effects similar to those of estrogen. There was in all groups a weak positive nuclear AR immunostaining. This is the first in vivo study on the effects on long-term hormonal treatment on the expression of PR isoforms in normal primate breast tissue. The results suggest that hormonal treatments have a different influence on the PRA/PRB balance in the breast.

Hormone receptor status in breast cancer--a comparison between surgical specimens and fine needle aspiration biopsies.

The present study was performed to evaluate the immunocytochemical analysis (ICA) of oestrogen (ER) and progesterone receptor (PR) in fine needle aspiration (FNA) biopsies from primary breast cancers as compared with the established enzyme immunoassays (ER-EIA and PR-EIA) based on cytosol homogenates from the corresponding resected tumour specimens. A total of 967 primary breast cancers were assessed for ER and PR content by both methods. Correlations between EIA and ICA expressed as percentage of tumour cells with a positive staining were highly significant (P < 0.001) for ER and PR. Staining intensity yielded only limited additional information. The concordance between the two techniques was about 80%. Evaluation of biological parameters by FNA may be useful to decide the optimal treatment for breast cancer patients.

The effect of long-term treatment with steroid hormones or tamoxifen on oestrogen receptors (alpha and beta) in the endometrium of ovariectomized cynomolgus macaques.

The effects of oestrogen are mediated by two specific intracellular receptors, oestrogen receptors (ER) alpha and beta, which function as ligand-activated transcriptional regulators. Ovariectomized macaques (Macaca fascicularis) were used to study the regulation of ERalpha and ERbeta in the endometrium by immunohistochemistry and in situ hybridization after long-term hormone treatment. Animals were treated continuously for 35 Months with either conjugated equine oestrogen (CEE), medroxyprogesterone acetate (MPA), combined CEE/MPA, or tamoxifen (TAM). Treatment with CEE/MPA down-regulated ERalpha in the superficial glands. In the superficial stroma the ERalpha level was lower in the CEE/MPA group than in the CEE and MPA groups. ERbeta immunostaining was faint with minor variation in response to treatment, but increased in the superficial stroma after MPA treatment. The ratio of ERbeta/ERalpha increased in superficial stroma and gland after CEE/MPA treatment, and also in stroma after MPA and TAM. Cystic endometrial hyperplasia was observed in TAM-treated animals, in combination with a high level of ERalpha protein expression. The present data show that long-term hormone treatment affects the ERalpha and ERbeta protein levels in the endometrium. The balance between ERalpha and ERbeta seems to be important for the proliferative response to oestrogen.

Expression of estrogen receptors (alpha, beta) and insulin-like growth factor-I in breast tissue from surgically postmenopausal cynomolgus macaques after long-term treatment with HRT and tamoxifen.

The novel estrogen receptor ERbeta could be a key factor for proliferation and breast cancer risk. In a primate model for long-term HRT, surgically postmenopausal cynomolgus macaques were treated for 35 months with conjugated equine estrogens (CEE), medroxyprogesterone acetate (MPA), CEE+MPA and tamoxifen (n=5 in all groups). The immunohistochemical expression of ERalpha, ERbeta and IGF-I in breast tissue was quantified by image analysis. Overall the levels of ERbeta were higher than for ERalpha. In untreated animals, the median area of positive cells was 58% and 21%. The lowest levels for ERbeta were seen during treatment with CEE/MPA (3%) and in this group the expression of ERbeta was lower than for ERalpha. Tamoxifen had effects similar to estrogen. ERbeta may have a role to modulate the proliferative response following activation of ERalpha. The results suggest that hormonal treatments have a different influence on the balance ERbeta/ERalpha in breast tissue.

Effects of oral contraceptives on breast epithelial proliferation.

The association between oral contraceptive (OC) use and breast cancer is not fully understood. Estrogen is a known mitogen to breast epithelial cells, but there is still a controversy about the effect of added progestogens. Fine needle aspiration (FNA) biopsies were used to assess epithelial proliferation in normal breast tissue from 106 healthy premenopausal women with and without oral contraceptives. In 26 women biopsies were performed before and after 2 months of OC use. Proliferation, expressed as percentage of Ki-67/MIB-1 positive cells, was correlated to endogenous progesterone, androgenic/anabolic compounds and exogenous progestogen. We found a higher proliferation (p = 0.03) in OC users compared to non users, with mean values of 4.8% and 2.2%, respectively. There was a positive correlation between proliferation and progesterone levels in non-users and with serum levonorgestrel concentrations in women using OCs containing this progestogen (rs = 0.43, p = 0.02). Women using OCs had significantly lower serum androgen levels compared to naturally cycling women and free testosterone levels displayed an inverse relation to breast epithelial proliferation. There was a marked variation in the response to exogenous sex steroids. In certain women after 2 months of OC use, the percentage of MIB-1 positive cells was as high as 40-50%. The results add to the growing evidence that progestogens may be mitogenic in breast tissue. Increased proliferation during hormonal contraception should be regarded as an unwanted and potentially hazardous side effect. Efforts should be made to define hormonal contraceptive regimens which minimize breast epithelial proliferation and to identify those women with the most pronounced proliferative response.

p53 expression in breast and endometrium during estrogen and tamoxifen treatment of surgically postmenopausal cynomolgus macaques.

Estrogens are important for both normal cell growth and malignant proliferation in the mammary gland as well as in the endometrium. Tamoxifen is a non-steroidal anti-estrogen widely used in breast cancer treatment. In recent years reports have been made of an increased risk of endometrial carcinoma during tamoxifen treatment. We used surgically menopausal cynomolgus macaques to study proliferation and p53 expression during hormonal replacement therapy (HRT) and tamoxifen treatment. Animals were treated continuously for 35 months with either conjugated equine estrogens (CEE; n = 20); medroxyprogesterone acetate (MPA; n = 17); the combination of CEE + MPA (n = 13); or tamoxifen (n = 17) for 35 months. We found an increased expression of p53 in normal breast and endometrial tissue linked to CEE but not tamoxifen treatment. In the breast alveoli there was an association between proliferation measured by morphometry and p53 expression in all groups. However, in the endometrium CEE induced significantly more p53 positivity than tamoxifen, 9/20 vs. 3/17 in glands and 9/19 vs. 0/17 in stroma, respectively. If indeed long-term treatment with tamoxifen as in the present study could inactivate the tumor-suppressive function of p53, endometrial cells might thereby become more susceptible to genetic lesions associated with carcinogenesis.

Expression of sex steroid receptors and IGF-1 mRNA in breast tissue--effects of hormonal treatment.

The mechanisms behind increased breast tissue proliferation and a possibly increased breast cancer risk in women using hormonal contraception (HC) and hormonal replacement therapy (HRT) are incompletely understood. We analyzed breast tissue from 20 premenopausal and seven postmenopausal women undergoing reduction mammoplasties for estrogen receptor (ER) and progesterone receptor (PR) content as well as mRNA levels for ER, PR and insulin-like growth factor-1 (IGF-1). The receptor values were correlated to IGF-1 mRNA concentrations and levels of steroid and peptide hormones and SHBG. In women using HC, we found significantly lower ER values (p = 0.02) but non-significantly lower ER mRNA levels compared to those in naturally cycling women. PR and PR mRNA were no different. Women on HC displayed a higher breast tissue proliferation (p = 0.05) expressed as Ki-67, MIB-1 positivity, which was correlated with IGF-1 mRNA (r(s) = 0.82, p = 0.04). Since the concentration of sex steroid receptors in breast tissue is comparatively low and steroid receptors are down-regulated during hormonal treatment, mechanisms other than direct sex steroid receptor action are likely to be present. Our results suggest a role for IGF-1 in the proliferative response of breast tissue during exogenous hormonal treatment.

Tamoxifen and secondary tumours. An update.

The nonsteroidal antiestrogen tamoxifen is the most widely used anticancer drug. In women with breast cancer, adjuvant therapy with tamoxifen reduces relapse and improves overall survival. In advanced breast cancer, the response rate is more than 50% in hormonal dependent disease. In women treated with adjuvant tamoxifen the incidence of new primary breast cancers is decreased. This latter observation has led to the initiation of prevention trials. In 1989 the first report from a large prospective randomised trial showed a significant increase of endometrial carcinoma among women treated with adjuvant tamoxifen. This effect may be linked to the somewhat paradoxical estrogenic properties of tamoxifen. The endometrial effects should be considered in the long term use of tamoxifen, and should also be taken into account in the evaluation of the prevention trials. Animal data indicate that tamoxifen can induce tumours in other organ systems, for example the liver, but no increase in primary liver cancer has been reported from the randomised trials. In some of these trials an increase in other gastrointestinal cancers (e.g. colon and gastric carcinoma) has been observed. The mechanism behind this may be different from that of the endometrium. In animal systems, tamoxifen has shown to induce DNA damage, with formation of DNA adducts. The risk of secondary gastrointestinal cancer needs to be further evaluated. The adverse effects of tamoxifen have led to the development of new anti-estrogenic drugs and other estrogen reducing agents (e.g. aromatase inhibitors).

Proliferation of breast epithelial cells in healthy women during the menstrual cycle.

OBJECTIVE: Our objective was to assess proliferation in normal breast epithelial cells from healthy women during the follicular and luteal phases of the menstrual cycle. STUDY DESIGN: We analyzed the proliferation marker Ki-67/MIB-1 by immunocytochemical methods in breast epithelial cells procured through fine needle aspiration biopsy from 47 healthy volunteers. Differences were assessed by Wilcoxon rank sum tests, and correlations were determined by Spearman's rank correlation coefficient. RESULTS: The proportion of KI-67/MIB-1-positive cells was higher in the luteal phase (2.04%) than in the follicular phase (1.66%). The values in women aged < 35 years were 2.29% and 1.13%, respectively (p = 0.003). In ovulating women with two aspirates during the same menstrual cycle the percentage of proliferating cells increased from the follicular phase (1.3%) to the luteal phase (2.4%) (p < 0.04). Proliferation was positively correlated with serum progesterone levels the day of aspiration (r = 0.34, p < 0.05). CONCLUSION: The fine needle aspiration biopsy technique is a valuable tool for in vivo studies of cell proliferation in the normal breast. Data clearly suggest a proliferative action of progesterone.

Blood clonal B-cell excess in patients with monoclonal gammopathy of undetermined significance (MGUS): association with malignant transformation.

Fifty-seven patients with monoclonal gammopathy of undetermined significance (MGUS) were analysed for the presence of blood clonal B-cell excess (CBE), defined as a lymphocyte surface membrane kappa/lambda light chain ratio outside the normal range (> 3.5 in kappa-type MGUS and < 0.9 in lambda-type MGUS). 15 patients (26%) had a CBE. The patients were followed for a median time of 8.4 years (range 0.5-20.2). Eight of the 15 CBE+ MGUS patients (53%) developed a B-cell malignancy as compared to 7/42 patients (17%) in the CBE group and the difference in event-free (malignancy-free) observation time was statistically significant (P = 0.01). Cox's regression analysis showed that the presence of CBE was the most powerful predictor of progression to a malignant disease. Two patients with a normal kappa/lambda ratio at first test were analysed repeatedly during follow-up. Subsequently, a CBE appeared which gradually increased in size preceding the clinical diagnosis of a malignant disease.