Effect of estradiol preceding and progesterone subsequent to ovulation on proportion of postpartum beef cows pregnant.

Two experiments were conducted to examine the effect of plasma concentrations of 17ß-estradiol (E2) preceding and progesterone (P4) subsequent to ovulation on proportions of beef cows pregnant following embryo transfer. Timing of ovulation (d 0) among postpartum cows was synchronized and cows that expressed estrus were removed from each study. In Experiment 1, plasma E2 concentration on d 0 was used to classify cows (n = 353) into Low, Medium, and High E2 groups. Pregnancy rate for cows with Low, Medium, or High E2 concentrations were different (P < 0.05). In Experiment 2, there were multiple administrations of PGF2α to evaluate the independent effects of Low or High E2 before ovulation and Low or Normal (no treatment) P4 after ovulation on proportions of cows pregnant. Treatment groups in Experiment 2, therefore, were: Low E2-Low P4 (LL; n = 71), Low E2-Normal P4 (LN; n = 69), High E2-Low P4 (HL; n = 74), and High E2-Normal P4 (HN; n = 73). Concentrations of P4 on d 7 subsequent to ovulation were less (P < 0.05) in cows of the HL compared to HN, and in LL compared to LN groups. Concentrations of E2 on d -2, 0, and change in E2 (d -2 to d 0) had a positive effect (P < 0.008) on pregnancy rates. In summary, relatively greater E2 concentrations preceding ovulation; and relatively greater P4 concentrations subsequent to ovulation combined with lesser E2 concentrations preceding ovulation had a positive effect on proportions of postpartum cows pregnant.

tRNA-derived fragments (tRFs) regulate post-transcriptional gene expression via AGO-dependent mechanism in IL-1ß stimulated chondrocytes.

OBJECTIVES: Recent studies have shown that tRNA-derived RNA fragments (tRFs) are novel regulators of post-transcriptional gene expression. However, the expression profiles and their role in post-transcriptional gene regulation in chondrocytes is unknown. Here, we determined tRFs expression profile and explored tRF-3003a role in post-transcriptional gene regulation in IL-1ß stimulated chondrocytes. METHODS: We used qPCR arrays to determine tRNAs and tRFs expression in age- and sex-matched primary human OA chondrocytes and TC28/I2 cells stimulated with IL-1ß. Chondrocytes were transfected with tRNA-CysGCA overexpression plasmid or tRF-3003a mimic and 3'UTR luciferase reporter plasmids of mRNAs harboring predicted tRF target "seed sequence". The AGO-RNA-induced silencing complex (AGO-RISC)-dependent repressive activity of tRF-3003a was determined by siRNA-mediated knockdown of AGO2. RESULTS: IL-1ß increased the expression levels of specific tRNAs and of tRF-3003a, a type 3 tRF produced by the cleavage of tRNA-CysGCA. tRF-3003a "seed sequence" was identified in the 3'UTR of JAK3 mRNA and tRNA-CysGCA overexpression or transfection of a tRF-3003a mimic in chondrocytes downregulated JAK3 expression and significantly reduced the activity of the 3'UTR reporter. RIP assay showed enrichment of tRF-3003a into AGO2/RISC in IL-1ß treated chondrocytes. The suppressive effect of tRF-3003a on JAK3 3'UTR reporter was abrogated with siRNA-mediated depletion of AGO2. CONCLUSIONS: We demonstrate that under pathological conditions chondrocytes display perturbations in the expression profile of specific tRNAs and tRFs. Furthermore, a specific tRF namely tRF-3003a can post-transcriptionally regulate JAK3 expression via AGO/RISC formation in chondrocytes. Identification of this novel mechanism may be of value in the design of precision therapies for OA.

Circulating concentrations of bovine pregnancy-associated glycoproteins and late embryonic mortality in lactating dairy herds.

The objectives of these experiments were as follows: (1) to determine the association between circulating concentrations of pregnancy-associated glycoproteins (PAG) and late embryonic mortality (EM) in lactating dairy cattle following fixed-time artificial insemination (TAI) on d 0 or timed embryo transfer (TET) on d 7, (2) to identify a circulating concentration of PAG on d 31 below which late EM would be likely to occur, and (3) to identify when during gestation (d 31-59) late EM is occurring. Cows were diagnosed pregnant on d 31 of gestation based on presence of a fetal heartbeat and reconfirmed to be pregnant on d 59 of gestation. Late EM occurred when a cow had a viable embryo on d 31 of gestation but not on d 59 following TAI or TET. Only pregnant cows on d 31 were included in the analysis (TAI-maintained, n=413; TAI-EM, n=77; TET-maintained, n=238; TET-EM, n=47). Cows that were pregnant at d 31 of gestation and maintained the pregnancy until d 59 had significantly higher circulating concentrations of PAG at d 31 of gestation compared with cows that experienced late EM between d 31 and 59 of gestation in both TAI and TET. To conduct a more stringent test of the effectiveness of a single circulating PAG concentration (d 31) to predict EM, a receiver-operating characteristic curve was generated to identify a PAG concentration on d 31 that would predict EM with ≥95% accuracy in cows that received TAI or TET. Based on positive and negative predicative value analysis, a circulating concentration of PAG below 1.4 ng/mL (TAI; minimal detectable level 0.28 ng/mL) and 1.85 ng/mL (TET) was 95% accurate in predicting EM (between d 31 and 59) at d 31 of gestation, respectively. Following TET, embryonic loss was tracked by Doppler ultrasound, progesterone, and PAG from d 24 to 59 of gestation, with more than 50% of the loss occurring between d 31 and 38 of gestation. In summary, circulating concentrations of PAG on d 31 of gestation may provide a good marker for predicting EM between d 31 and 59 of gestation, and the data suggest that this model could help predict which cows will undergo late EM.

Weekly AUC2 carboplatin in acquired platinum-resistant ovarian cancer with or without oral phenoxodiol, a sensitizer of platinum cytotoxicity: the phase III OVATURE multicenter randomized study.

BACKGROUND: Platinum-resistant ovarian cancer (PROC) constitutes a therapeutic dilemma with limited efficacy from traditional cytotoxic agents. Based on prior data suggesting that scheduling alterations of platinum would increase activity, the aim of the present study was to assess the potential therapeutic benefit of phenoxodiol (PXD), a novel biomodulator shown to have chemoresistance reversing potential, when combined with weekly AUC2-carboplatin in PROC patients. PATIENTS AND METHODS: A multicenter randomized double-blind placebo controlled phase-III-study was conducted to compare oral PXD plus AUC2-carboplatin (group 1) versus placebo plus AUC2-carboplatin (group 2) weekly in PROC patients. The primary end point was progression-free-survival (PFS). Secondary objectives included overall survival (OS), response rates, duration of response and quality of life. RESULTS: The study was terminated early 14 April 2009, after recruitment of 142 patients due to feasibility and recruitment challenges. A total of 142 patients were randomized. The groups were well balanced in terms of important baseline characteristics. The median PFS for group 1 was 15.4 weeks [95% confidence interval (CI) 11.1-21.0] versus 20.1 weeks for group 2 (95% CI = 13.1-33.4); P = 0.3. The objective response rate and median survival in group 1 versus group 2 was 0% versus 1% and 38.3 weeks (95% CI 32.0-45.3) versus 45.7 weeks (95% CI 35.6-58.0), respectively. PXD appeared to be well tolerated. The main reason for dose modification in both groups was hematologic toxicity. CONCLUSIONS: Orally delivered PXD showed no evidence of clinical activity, when combined with weekly AUC2-carboplatin in PROC. In addition, single-agent weekly AUC2-carboplatin appeared to be inactive by response criteria in a homogenously defined population of PROC. This has implications for the design of future studies.

Circulating bovine pregnancy associated glycoproteins are associated with late embryonic/fetal survival but not ovulatory follicle size in suckled beef cows.

The objective was to examine the relationship between ovulatory follicle size and embryo and fetal survival by using circulating concentrations of bovine pregnancy associated glycoproteins (bPAG) to detect the presence of an embryo or fetus and monitor placental function. Before examining the relationship between bPAG, ovulatory follicle size, and embryo and fetal survival, the half-life of bPAG was determined in Exp. 1. The half-life of bPAG after PGF2α-induced abortion on d 32 to 36 postinsemination was 35.8 ± 21.9 h (mean ± SD; range 7.1 to 78.5 h). In Exp. 2, suckled beef cows (n = 91) were treated with the CO-Synch protocol (GnRH on d -9, PGF2α on d -2, and GnRH and AI 48 h later [d 0]) and classified into 1 of 2 ovulatory follicle size groups: 1) small follicle (<12.5 mm; n = 25) or 2) large follicle (≥ 12.5 mm; n = 66). The first increase (P < 0.0001) in serum bPAG occurred in pregnant cows on d 24 after insemination and circulating bPAG decreased before a decrease in progesterone in 3 of 4 cows that lost an embryo or fetus. Pattern of secretion of bPAG in serum from d 24 to 60 after insemination (d 0) was affected by day (P < 0.0001), but not ovulatory follicle size. In Exp. 3, suckled beef cows (n = 1164) were administered the CO-Synch protocol either with (donor cows; n = 810) or without (recipient cows; n = 354) AI on d 0. Single embryos (n = 394) or oocytes (n = 45) were recovered from the donor cows [d 7; embryo transfer (ET)] and all live embryos were transferred into recipients the same day. Cows were classified on d 0 as having a small (<12.5 mm) or large (≥ 12.5 mm) ovulatory follicle, and randomly chosen as donors or recipients to remove confounding effects of ovulatory follicle size on fertility. Serum concentration of bPAG at d 28 was not affected by ovulatory follicle size (P = 0.85), embryo stage at ET (P = 0.75), embryo quality at ET (P = 0.64), estradiol at GnRH2 (P = 0.62) or serum progesterone at ET (d7; P = 0.14). Compared with cows that maintained pregnancy (n = 176), cows that exhibited late embryonic or fetal mortality (n = 19) after d 28 had decreased (P < 0.05) concentrations of bPAG on d 28. In summary, there was no relationship between serum bPAG and ovulatory follicle size or embryo stage or quality at ET; however, cows that lost an embryo after d 28 had reduced concentrations of bPAG on d 28 compared with cows that maintained pregnancy.

The mutational profile of sporadic epithelial ovarian carcinoma.

Mutations occurring in sporadic epithelial ovarian carcinomas are reviewed and their functional significance in terms of prognosis and prediction of anticancer drug activity are discussed. Alterations in the BRCA1/2 genes, TP53, PTEN, PI3Kinase, KRAS/BRAF and CTNNB1 are described. TP53 is likely to be a driver in high grade serous tumours, but is less useful than BRCA status in prediction of response to the platinum or PARPi agents. It is expected that mutation profiling will become integrated into current morphological/immunohistochemical primary diagnostic assessment of tumours once the cost and quality control issues of the technology are addressed.

Salivary gland gene expression atlas identifies a new regulator of branching morphogenesis.

During organ development, local changes in gene expression govern morphogenesis and cell fate. We have generated a microanatomical atlas of epithelial gene expression of embryonic salivary glands. The mouse submandibular salivary gland first appears as a single mass of epithelial cells surrounded by mesenchyme, and it undergoes rapid branching morphogenesis to form a complex secretory organ with acini connected to an extensive ductal system. Using laser capture microdissection, we collected samples from 14 distinct epithelial locations at embryonic days 12.5, 13.5, 14, and 15, and characterized their gene expression by microarray analysis. These microarray results were evaluated by qPCR of biological replicates and by comparisons of the gene expression dataset with published expression data. Using this gene expression atlas to search for novel regulators of branching morphogenesis, we found a substantial reduction in mRNA levels of GSK3ß at the base of forming clefts. This unexpected finding was confirmed by immunostaining, and inhibition of GSK3ß activity enhanced salivary gland branching. This first microanatomical expression atlas of a developing gland characterizes changes in local gene expression during salivary gland development and differentiation, which should facilitate the identification of key genes involved in tissue morphogenesis.

Effects of resynchronization programs on pregnancy per artificial insemination, progesterone, and pregnancy-associated glycoproteins in plasma of lactating dairy cows.

Objectives were to develop a timed artificial insemination (TAI) resynchronization program to improve pregnancy per AI and to evaluate responses of circulating progesterone and pregnancy-associated glycoproteins in lactating cows. Cows (n=1,578) were presynchronized with 2 injections of PGF2alpha, given 14 d apart starting on d 45+/-3 postpartum, followed by Ovsynch [2 injections of GnRH 7 d before and 56 h after injection of PGF2alpha, TAI 16 h after second injection (d 0)]. The Resynch-treated cows received an intravaginal progesterone insert from d 18 to 25, GnRH on d 25, and pregnancy diagnosis on d 32, and nonpregnant cows received PGF2alpha., GnRH 56 h later, and TAI 16 h later (d 35). The control cows were diagnosed for pregnancy on d 32 and nonpregnant cows received GnRH, PGF2alpha 39 d after TAI, GnRH 56 h later, and TAI 16 h later (d 42). Pregnancy was reconfirmed on d 60 after AI. Ovarian structures were examined in a subset of cows at the time of GnRH and PGF2alpha injections. Blood samples for analyses of progesterone and pregnancy-associated glycoproteins were collected every 2 d from d 18 to 30 in 100 cows, and collection continued weekly to d 60 for pregnant cows (n=43). Preenrollment pregnancies per AI on d 32 did not differ for cows subsequently treated as Resynch (45.8%, n=814) and control (45.9%, n=764), and pregnancy losses on d 60 were 6.7 and 4.0%, respectively. Resynchronized service pregnancy per AI (36%, n=441; 39.5%, n=412) and pregnancy losses (6.3 and 6.7%) did not differ for Resynch and control treatments, respectively. Days open for pregnant cows after 2 TAI were less for the Resynch treatment than for the control treatment (96.2+/-0.82 vs. 99.5+/-0.83 d). Cows in the Resynch treatment had more large follicles at the time of GnRH. The number of corpora lutea did not differ between treatments at the time of PGF2alpha. Plasma progesterone for pregnant cows was greater for Resynch cows than for control cows (18-60 d; 6.6 vs. 5.3 ng/mL), and plasma concentrations of progesterone on d 18 were greater for pregnant cows than for nonpregnant cows (5.3 vs. 4.3 ng/mL). Plasma pregnancy-associated glycoproteins during pregnancy were lower for cows in the Resynch treatment compared with control cows on d 39 (2.8 vs. 4.1 ng/mL) and 46 (1.3 vs. 3.0 ng/mL). Cows pregnant on d 32 that lost pregnancy by d 60 (n=7) had lower plasma concentrations of pregnancy-associated glycoproteins on d 30 than cows that maintained pregnancy (n=36; 2.9 vs. 5.0 ng/mL). Pregnancy-associated glycoproteins on d 30 (>0.33 ng/mL) were predictive of a positive d 32 pregnancy diagnosis (sensitivity=100%; specificity=90.6%). In conclusion, Resynch and control protocols had comparable pregnancy per AI for first and second TAI services, but pregnancy occurred 3.2 d earlier in the Resynch group because inseminations in the Resynch treatment began 7 d before those in the control treatment. Administration of an intravaginal progesterone insert, or GnRH, or both increased progesterone during pregnancy. Dynamics of pregnancy-associated glycoproteins were indicative of pregnancy status and pregnancy loss.

Platinum-based adjuvant chemotherapy for early-stage epithelial ovarian cancer: single or combination chemotherapy?

OBJECTIVE: To evaluate the clinical benefit and toxicity of two regimens; single agent carboplatin (C) and a carboplatin/paclitaxel (CP) combination in early epithelial ovarian cancer. DESIGN: A retrospective review. SETTING: Single cancer centre serving a population of 2.1 million in the northwest of England. POPULATION: All women treated with adjuvant chemotherapy for International Federation of Obstetrics and Gynecology stage Ia-IIc ovarian cancer between 2002 and 2005. METHODS: Case and operation notes were reviewed. Details of the surgery performed, performance status (PS), tumour histology, stage, grade, intended chemotherapy, chemotherapy received, acute and late toxicity, relapse and death were all recorded. MAIN OUTCOME MEASURES: Overall survival (OS), relapse-free survival (RFS), acute and late toxicity. RESULTS: Sixty women received CP and 35 received C. Younger women (P < 0.0001) and those with a better performance status (P = 0.045) were more likely to receive CP. Median follow- up was 38 months (range 0-70). Five-year OS was 62% (95% CI 44-81%) for C and 73% (95% CI 61-85%) for CP P= = 0.316. For the subgroup with stage I disease and good PS (0/1) 5-year OS was 80% (59-100%) for C and 79% (63-95%) for CP; P = 1.0. For those with stage 2 disease, 5-year OS was 29% (95% CI 0-62%) for C and 63% (95% CI 44-82%) for CP; P = 0.025. Subgroup analyses by grade or histology showed no difference in OS. P was discontinued prematurely in nine (15%) women on account of toxicity, whereas C was not stopped early. P-related neuropathy (G1/2) was reported in ten (17%) women at 6 months and in two (3%) at 1 year. CONCLUSIONS: Combination therapy is administered more often than carboplatin; especially in those with younger age, better PS and nonmucinous histology. Recurrence and death rates were similar with both treatments. Well-designed trials are needed to identify the optimum chemotherapy regimen in this group.

Measurement of the ratio of the numu charged-current single-pion production to quasielastic scattering with a 0.8 GeV neutrino beam on mineral oil.

Using high statistics samples of charged-current numu interactions, the MiniBooNE [corrected] Collaboration reports a measurement of the single-charged-pion production to quasielastic cross section ratio on mineral oil (CH2), both with and without corrections for hadron reinteractions in the target nucleus. The result is provided as a function of neutrino energy in the range 0.4 GeV

Dietary protein during gestation affects circulating indicators of placental function and fetal development in heifers.

The influences of nutritional protein during the first and second trimesters of pregnancy on placental hormones and fetal growth were determined in composite beef heifers. At artificial insemination, heifers were stratified by weight within each composite genotype into 4 treatment groups: High High (HH=1.4kg crude protein (CP)/day for first and second trimesters of gestation; n=16), High Low (HL=1.4kg CP/day for first trimester and 0.4kg CP/day for second trimester; n=19), Low High (LH=0.4kg CP/day for first trimester and 1.4kg CP/day for second trimester; n=17) or Low Low (LL=0.4kg CP/day for first and second trimesters; n=19). Maternal plasma bovine pregnancy associated glycoprotein (bPAG) and progesterone (P4) were determined at gestation day (gd) 28, 82, 179 and 271 (mean gestation length 286 days) in addition to P4 at term. Estrone sulphate (ES) and bovine placental lactogen (bPL) concentrations were measured at gd 124, 179, 236 and 271 and at term in addition to ES at gd 82. Low dietary protein increased placental function as indicated by increased bPAG (P<0.001) and ES (P=0.02) concentrations in first trimester and increased bPL concentrations (P=0.01) in the second trimester of gestation. In the third trimester, when dietary treatment had ceased, placental function was no longer associated with previous dietary treatments. Dam genotype affected placental function as measured by bPL (P<0.001) and ES concentrations (P=0.02). Calf gender, heifer age and maternal insulin-like growth factor (IGF)-I, -II and leptin did not affect hormonal indicators or circulating markers of placental function. Enhanced placental function during the third trimester, as measured by ES, was associated with increased calf birth weight (P=0.003).

Antibody-mediated enhancement of the rate, magnitude, and responsiveness of vesicular stomatitis virus induced alpha interferon production.

A majority of adults without evidence of exposure to vesicular stomatitis virus (VSV) have serum IgG antibodies that interact with pro-inflammatory TLR7 in the presence of VSV, and enhance several aspects of VSV-induced IFN-alpha production. Enhancing IgG antibody enables human PBMC to make IFN-alpha more rapidly and in higher titers in response to a broad range of VSV-concentrations that include those too low to independently stimulate IFN-alpha production. These antibody-mediated functions compensate for the inherent delay in virus-induced IFN-alpha production in vitro, and have the potential to improve the in vivo IFN-alpha response and effectively terminate infection before the occurrence of clinically apparent disease. The frequent presence of enhancing antibody in persons without predictable VSV exposure has implications for naturally occurring infections with this and other viruses, and for the use of viruses as vaccine vectors and oncolytic agents.

Promoter hypermethylation of FANCF and outcome in advanced ovarian cancer.

The Fanconi gene family has a role in DNA repair and inactivation of FANCF has been proposed as a mechanism of sensitisation to platinum chemotherapy. This study sought to confirm this hypothesis in cell lines and a large series of ovarian cancer samples. Promoter methylation was assessed by methylation-sensitive polymerase chain reaction of FANCF in nine ovarian cancer cell lines and 74 ovarian cancer samples taken from patients entered on a trial of cisplatin-based chemotherapy. This study confirmed methylation-dependent silencing of FANCF in one out of nine ovarian cancer cell lines. Methylation of FANCF was demonstrated in 13.2% of 53 evaluable ovarian tumour samples. Progression-free survival gave an HR of 3.63 (95% CI: 1.54-8.54, P=0.0016) in favour of the unmethylated cases. There was no association with overall survival. This study does not support methylation-dependent silencing of FANCF as a mechanism of sensitisation to platinum-based chemotherapy in ovarian cancer.

Identification of survivin, an inhibitor of apoptosis, in canine urinary bladder transitional cell carcinoma.

Survivin, an inhibitor of apoptosis, is overexpressed in human invasive transitional cell carcinoma (TCC) of the urinary bladder. Survivin expression in canine TCC has not been defined. This study was designed to compare survivin expression between canine TCC and normal urinary bladder tissue. Reverse-transcriptase polymerase chain reaction (PCR) and immunohistochemistry (IHC) were performed on fresh-frozen and formalin-fixed tissues, respectively. All TCC tissues (n = 6) and 11/22 normal tissues assessed by PCR were positive for survivin. This difference was not significant (P = 0.06). With regard to IHC, 28/41 TCC samples were positive for nuclear survivin, whereas 0/46 normal tissues had nuclear immunoreactivity (P < 0.001). Cytoplasmic immunoreactivity did not significantly differ between TCC (7/41) and normal tissues (17/46) (P = 0.07). We conclude that nuclear survivin is present in canine TCC, but not in normal bladder urothelium. Future studies will evaluate the role of nuclear survivin in TCC development and as a potential therapeutic target.

Pediatric ERCP in a multidisciplinary community setting: experience with a fellowship-trained general surgeon.

UNLABELLED: Endoscopic retrograde cholangiopancreatography (ERCP) has been used to evaluate and treat pancreaticobiliary disorders and trauma in the pediatric population. Still representing a small percentage of total pediatric endoscopies, this procedure has been performed most commonly by a small subset of adult and pediatric gastroenterologists at quaternary referral centers. METHODS: In this study, we present a review of one fellowship-trained general surgeon's experience with pediatric ERCP in a teaching community pediatric hospital for the purpose of comparison with national series. RESULTS: All ERCPs performed by one general surgeon as part of a multidisciplinary team over a 5-year period in patients aged 16 years or less were reviewed. Success and complication rates were compared between our series and published pediatric and adult series using Fisher's exact test. Comparisons were made of indications, type of anesthesia, final diagnosis, and therapeutic interventions to ensure similar study populations. A total of 26 ERCPs were performed in 19 patients ranging from 7 to 16 years old. Therapeutic procedures included sphincterotomy (11), stent placement (7), stone removal (3), and dilation (2). In one case, stone removal and stent placement were performed in conjunction with pancreatic lithotripsy. In two cases the involved duct was not visualized. There were no instances of pancreatitis, bleeding, or perforation related to ERCP. CONCLUSIONS: When compared with published series, our data demonstrated no significant difference in success or complication rates. Our study demonstrates that pediatric ERCP can be performed by fellowship-trained general surgeons with success and complication rates comparable to accepted standards. Integration of the ERCP-trained general surgeon into the pediatric team is a potential asset in the care of pediatric patients with pancreaticobiliary disorders.

Hormone therapy in advanced and recurrent endometrial cancer: a systematic review.

Endometrial cancer is a hormone-dependent malignancy, and the majority has a precursor phase of endometrial hyperplasia. Histologic subtypes have been recognized with differing natural history. The relationship between hormone response, histology, and molecular profile is not established, but the relevant biology is summarized. This study was a systematic review of the literature to identify which populations should be considered for hormone interventions. Systematic searches were carried out in the English literature for randomized controlled trials and phase II studies of hormone interventions in endometrial cancer. Five randomized trials and 29 phase II studies were identified comprising a total of 2471 patients. In previously untreated patients with grade 1 (G1) or G2 tumors, the response rate for progestogens and the progression-free survival is in the range of 11-56% and 2.5-14 months, respectively. Higher response rates are seen in progesterone receptor-positive cases. Phase II studies comprise the majority of the data and many are of poor quality. There was considerable heterogeneity in patient selection, prior treatment, and type of regimen, and meta-analysis was not possible. G3 or G4 toxicity was less than 5%. We conclude that hormone receptor assessments should be carried out in all patients entered on clinical trials and may aid clinical management in selected cases. Receptor-negative status should not be an absolute contraindication to hormone intervention. Integration of hormone treatment with conventional chemotherapy and growth factor-targeted therapy needs to be explored.