Autoantibodies (AAb) directed against the nuclear phosphoprotein p53 can be detected in patients with various forms of cancer. The objective was to determine the prevalence of p53 AAb at the time of diagnosis in ovarian cancer patients and to correlate the presence of p53 AAb with clinicopathological parameters. Sera of 83 patients were analyzed by an ELISA using p53 expressed from a human wild-type cDNA. p53 AAb were detectable at all stages. The overall prevalence was 46%. p53 AAb were more frequent in patients with higher age (p = 0.014), postmenopausal status (p = 0.050), or advanced tumor stage (p = 0.046). p53 AAb positivity was related to the proportion of cells positive in immunohistochemistry but not with the staining intensity. In bivariate analysis, patients with p53 AAb had a 1.96-fold risk for relapse (95% confidence interval 1.02–3.78).

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The fetus is supplied from the placenta with estradiol (E2) and progesterone (P) in increasing amounts during gestation. After delivery of a premature infant, placental supply is disrupted, resulting in a rapid decrease in E2 and P. Replacement of these placental hormones may restore intrauterine conditions and may be beneficial for bone mineral accretion, clinical course, and outcome. Thirty female infants with a median gestational age of 26.6 weeks (between 24.1-28.7) and a birth weight of 675 g (370-990) were randomized to receive E2 and P replacement, aiming to maintain plasma levels equaling the intrauterine levels, or no replacement. The E2 and P replacement was started iv and was followed by transepidermal administration for a total duration of 6 weeks. Repeated measurements included plasma levels of E2, P, FSH, and LH; uterine volume; calcium and phosphorus in spot urine specimens; and bone mineral accretion by single photon absorption densitometry. Further, the incidence of chronic lung disease and various clinical outcome data were recorded. The plasma levels of E2 and P were within the intrauterine range with median replacements of 2.30 mg/kg x day E2 (1.13-6.23) and 21.20 mg/kg x day P (11.23-27.36), iv. Three- and 6-fold higher doses of E2 and P were needed via the transepidermal route. The uterine volumes increased, and FSH and LH as indicators for biological effectiveness were significantly lowered with replacement. The bone mineral accretion rates tended to be higher, and the incidence of chronic lung disease tended to be lower (0% vs. 29%; P = 0.097). E2 and P replacement via iv and transepidermal routes is capable of maintaining plasma levels as high as those in utero with biological effectiveness. Trends toward improved postnatal bone mineral accretion and less chronic lung disease were found with the hormone replacement. Further and more extensive studies are warranted to address the role of this new approach in the care of extremely premature infants.

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SummaryThe prognostic value of disseminated tumour cells derived from 353 breast cancer patients was evaluated. Disseminated tumour cells were purified from blood using a newly established method and nucleic acids were subsequently isolated. We investigated genomic imbalances (GI) such as mutation, amplification and loss of heterozygosity of 13 tumour suppressor genes and 2 proto-oncogenes using DNA from isolated minimal residual cancer cells. Significant correlations were found between genomic alterations of the DCC-and c-erbB-2 genes in disseminated breast cancer cells and actuarial relapse-free survival. Furthermore, increasing numbers of genomic imbalances measured in disseminated tumour cells were significantly associated with worse prognosis of recurrent disease. Logistic regression and Cox multivariate analysis led to the identification of genomic imbalances as an independent prognostic factor. Determination of disseminated tumour cells by genotyping of oncogenes and tumour suppressor genes seems not only to be a useful adjunct in follow up of carcinoma patients but provides also valuable additional individualized prognostic and predictive information in breast cancer patients beyond the TNM system.

p53 Genetic alterations are associated with advanced stage and aggressive tumors in a variety of human malignancies. The aim of this study was to examine p53 for genetic alterations and to evaluate the association of these alterations with clinical outcome and response to adjuvant radiotherapy in endometrioid endometrial carcinomas. p53 mutations in exons 2-11 were assessed in 59 endometrioid carcinomas by polymerase chain reaction-single-strand conformational polymorphism and sequence analysis. Twelve mutations (20.3%) and nine polymorphisms were identified. Seven of the nine polymorphisms were codon 72 single nucleotide polymorphisms (SNP) with an Arg/Pro allelotype. Women harboring either a mutation or an Arg/Pro allelotype at codon 72 had a lower overall survival rate than women whose tumors lacked alterations in the p53 gene (P= 0.0029). Women were stratified based on p53 genetic alterations (p53 mutation or p53 codon 72 SNP) and whether or not they received adjuvant radiation therapy. Women with p53 genetic alterations who did not receive adjuvant radiotherapy had the lowest survival rate (P= 0.0005). Treated women with p53 genetic alterations and untreated women with no p53 alteration had similar rates of survival. Among women with p53 alterations, adjuvant radiotherapy substantially increased survival (P= 0.035). In multivariate analyses, the group of women with p53 genetic alterations who did not receive adjuvant radiation therapy had a 5.9-fold increased risk of death (95% confidence interval: 1.5-22.7) compared to women whose tumors lacked p53 alterations and did not receive adjuvant radiation therapy.

During pregnancy, 17beta-estradiol (E2) and progesterone (P) plasma concentrations increase up to 100-fold. The fetus is exposed to these increasing amounts of E2 and P. Within 1 d after delivery, E2 and P concentrations fall to nonpregnancy concentrations in the mother and the infant. Extremely premature infants are cut off from the placental supply of E2 and P at a very early developmental stage, and therefore they suffer from this deprivation for a longer period than infants born at term. Nothing is known about the consequences of this deprivation. The purpose of this study was to investigate how intrauterine concentrations of E2 and P could be maintained after birth. In 13 infants with a median gestational age of 26.4 wk (24.1-28.7), a phospholipid-stabilized soybean oil emulsion available for parenteral nutrition that contains different amounts of E2 and P was continuously administered, starting within the first postnatal hours. The supplementation was continued as long as venous access was indicated but not longer than 6 wk (median 20 d, 12-44). To maintain intrauterine plasma concentrations of 2000-6000 pg/mL E2 and 300-600 ng/mL P, 2.30 mg x kg(-1) x d(-1) E2 (1.13-3.42 mg x kg(-1) x d(-1)) and 21.20 mg x kg(-1) x d(-1) P (11.23-27.36 mg x kg(-1) x d(-1)) were needed. We conclude that supplementation of E2 and P to maintain intrauterine concentrations in extremely premature infants is possible intravenously. The infants in this study are enrolled in a randomized, controlled pilot study to evaluate the potential benefits of E2 and P supplementation.

A new, general methodology for 'sandwich affinity chromatography of steroid hormone receptors is proposed, the part purification of the human spleen tumor glucocorticoid receptor is quoted as an illustration. 9-Fluoro-I 6~-methyl-1 l/?,l7-dihydroxy-l,4-androstadiene-3-one-l7~~-carboxylic acid was coupled to biotin using pentamethylenediamine (BioDex 1) as a spacer. The bifunctional derivative binds to glucocorticoid receptors and avidinSepharose and efficiently protects the glucocorticoid receptor against inactivation when previously added during homogenisation. We have standardized the capacity and optimum conditions for elution of receptor-BioDex-I complexes which are bound to avidin-Sepharose. Receptor purification of several thousandfold can be obtained with good yieldIncreasing attention has been directed recently towards the development of new affinity chromatography methods for hormone receptor purification [l, 21. The most promising method for purification of these proteins seems to be using hormone-biotin conjugates which retain their high binding constant towards avidin when bound to the respective hormone receptor.Bifunctional glucocorticoid-biotin analogs added during tissue homogenization are not only able to form stable hormone-receptor complexes but also retain their high specificity for immobilized avidin [2]. We have synthesized a number of steroid derivatives which contain biotin covalently attached to the steroid side-chain. This report describes the synthesis of biotin-labelled dexamethasone. Its properties appear to satisfy the basic criteria required of a good affinity probe. EXPERIMENTAL PROCEDURES Hormones and Chc~micalsand [5'-3H]thyniidine (5 Ciimmol) were purchased from the Amcrsham International, England. Biotin was purchased from Sigma Munchen, FRG. All other chemicals were purchased from Merck Darmstadt, F R G and were of analytical grade.The scheme for synthesis of BioDex 3 -3 is outlined in Fig. 1 In the second step of synthesis, 40mg of the active ester derivative 11 were dissolved in 50 ml of ethyl acetate, 100 pl of pentamethylenediamine were added, and the mixture was allowed to react at room temperature for 16 h. The organic solvent was thoroughly washed with water and removed on a rotary evaporator. The remaining residue was redissolved in benzene and lyophilized. The amine produced one spot on TLC in solvent systems A and B, respectively and was used without further purification.The synthesis of amino compoundsIV and V was performed as described above except that ethylenediamine (IV) and cystamine (V) were used. BioDex 1. ['4C]Biotinyl-N-hydroxysuccinimide ester (specific activity 0.2 mCi I4C/g biotin), as well as the unlabelled ester, were prepared from biotin and N-hydroxysuccinimide by coupling with dicyclohexylcarbodiimide. In the next step of synthesis, 20 mg of the amine 111 were dissolved in lOmi of ethanol, 29 mg of the biotin reagent (twofold excess) were added and the mixture was allowed to react overnight at room temperature. The crude product was purified by pre...