Search

We investigate some aspects of the problem of the estimation of birth distributions (BD) in multi-type Galton-Watson (MGW) trees with unobserved types. More precisely, we consider a two-type MGW called spinal-structured trees. This kind of tree is characterized by a spine of special individuals whose BD $\nu$ is different from the other individuals in the tree (called normal whose BD is denoted $\mu$). In this work, we show that even in such a very structured two-types population, our ability to distinguish the two types and estimate $\mu$ and $\nu$ is constrained by a trade off between the growth-rate of the population and the similarity of $\mu$ and $\nu$. Indeed, if the growth-rate is too large, large deviations events are likely to be observed in the sampling of the normal individuals preventing us to distinguish them from special ones. Roughly speaking, our approach succeed if $r, Comment: 27 pages

Resume L’embolie pulmonaire est une maladie grave et potentiellement mortelle. Son incidence chez les enfants drepanocytaires est probablement sous-estimee et les cas rapportes dans la litterature, notamment en pediatrie, sont rares. Par ailleurs, les symptomes cliniques peuvent mimer un syndrome thoracique aigu. Nous rapportons le cas d’un adolescent drepanocytaire de 17 ans admis pour une douleur thoracique avec dyspnee et tachycardie. L’angioscanner pulmonaire a revele une embolie pulmonaire bilaterale partiellement obstructive. Le bilan etiologique n’a pas mis en evidence de thrombus veineux profond ni de thrombophilie. L’evolution a ete rapidement favorable sous traitement anticoagulant. Nous preconisons la realisation d’un angioscanner pulmonaire devant toute douleur thoracique n’evoluant pas favorablement chez un enfant drepanocytaire. Le recensement d’autres cas pediatriques sera utile afin d’etablir des recommandations diagnostiques et therapeutiques.

Pulmonary embolism is a life-threatening and potentially lethal disease. Its incidence in children with sickle cell disease is probably underestimated and pediatric case reports in the literature are rare. Moreover, symptoms can mimic an acute chest syndrome. We report on the case of a 17-year-old boy with SS sickle cell disease, admitted for chest pain with dyspnea and tachycardia. Pulmonary angiography revealed a partial bilateral obstructive pulmonary embolism. We did not find any deep venous thrombosis or thrombophilia. The progression was rapidly favorable with anticoagulant therapy. We recommend a pulmonary angiography for any chest pain that does not evolve favorably in a child with sickle cell disease. Large series of pediatric patients would be useful to establish diagnostic and therapeutic guidelines.

Les proteases jouent un role crucial dans la reparation tissulaire et le processus de croissance et invasivite tumorale. Cependant sur le plan physiopathologique, la quasi totalite de ces enzymes proteolytiques sont secretees sous forme de proproteases inactives. A l’exception de la cathepsine D qui est capable de s’autoactiver, l’ensemble des autres proteases sont activees par un mecanisme commun a partir de la plasmine, elle-meme derivant de la transformation du plasminogene, pro molecule ubiquitaire. uPA est l’initiateur de la transformation activatrice du plasminogene en plasmine. Son recepteur specifique uPAR est un recepteur de surface extra membranaire, sans domaine transmembranaire ni relie a des voies de transduction du signal intracellulaires. uPAR est cependant capable de s’heterodimeriser avec d’autres molecules membranaires (recepteurs des facteurs de croissance, molecules d’adhesion cellulaire....). Ces molecules transmembranaires, apres heterodimerisation avec uPAR, activent les voies de transduction du signal qui leur sont associees. uPAR est indispensable pour l’activation de uPA, mais egalement pour la mobilisation cellulaire en association avec le complexe integrines αvβ3. L’heterodimerisation uPAR - αvβ3 se fait au front anterieur des lamellipodes des cellules mobilisables (cellules epitheliales cancereuses EMT positives, cellules endotheliales activees, macrophages associes aux tumeurs, neofibroblastes...). PAI1, inhibiteur actif de l’activite uPA, a un role complexe dans la tumorigenese

Recently, clustering moving object trajectories kept gaining interest from both the data mining and machine learning communities. This problem, however, was studied mainly and extensively in the setting where moving objects can move freely on the euclidean space. In this paper, we study the problem of clustering trajectories of vehicles whose movement is restricted by the underlying road network. We model relations between these trajectories and road segments as a bipartite graph and we try to cluster its vertices. We demonstrate our approaches on synthetic data and show how it could be useful in inferring knowledge about the flow dynamics and the behavior of the drivers using the road network.

For the last few years, the amount of data has significantly increased in the companies. It is the reason why data analysis methods have to evolve to meet new demands. In this article, we introduce a practical analysis of a large database from a telecommunication operator. The problem is to segment a territory and characterize the retrieved areas owing to their inhabitant behavior in terms of mobile telephony. We have call detail records collected during five months in France. We propose a two stages analysis. The first one aims at grouping source antennas which originating calls are similarly distributed on target antennas and conversely for target antenna w.r.t. source antenna. A geographic projection of the data is used to display the results on a map of France. The second stage discretizes the time into periods between which we note changes in distributions of calls emerging from the clusters of source antennas. This enables an analysis of temporal changes of inhabitants behavior in every area of the country., Advances in Knowledge Discovery and Management, 615, Springer International Publishing, pp.3-17, 2015, Studies in Computational Intelligence

We suggest a novel method of clustering and exploratory analysis of temporal event sequences data (also known as categorical time series) based on three-dimensional data grid models. A data set of temporal event sequences can be represented as a data set of three-dimensional points, each point is defined by three variables: a sequence identifier, a time value and an event value. Instantiating data grid models to the 3D-points turns the problem into 3D-coclustering. The sequences are partitioned into clusters, the time variable is discretized into intervals and the events are partitioned into clusters. The cross-product of the univariate partitions forms a multivariate partition of the representation space, i.e., a grid of cells and it also represents a nonparametric estimator of the joint distribution of the sequences, time and events dimensions. Thus, the sequences are grouped together because they have similar joint distribution of time and events, i.e., similar distribution of events along the time dimension. The best data grid is computed using a parameter-free Bayesian model selection approach. We also suggest several criteria for exploiting the resulting grid through agglomerative hierarchies, for interpreting the clusters of sequences and characterizing their components through insightful visualizations. Extensive experiments on both synthetic and real-world data sets demonstrate that data grid models are efficient, effective and discover meaningful underlying patterns of categorical time series data.

A piecewise-deterministic Markov process is a stochastic process whose behavior is governed by an ordinary differential equation punctuated by random jumps occurring at random times. We focus on the nonparametric estimation problem of the jump rate for such a stochastic model observed within a long time interval under an ergodicity condition. We introduce an uncountable class (indexed by the deterministic flow) of recursive kernel estimates of the jump rate and we establish their strong pointwise consistency as well as their asymptotic normality. We propose to choose among this class the estimator with the minimal variance, which is unfortunately unknown and thus remains to be estimated. We also discuss the choice of the bandwidth parameters by cross-validation methods., 36 pages, 18 figures

Objectif Le 25I-NBOMe, hallucinogene de synthese quelquefois vendu comme du LSD, est disponible sur Internet depuis plus de 5 ans. Ingere, sniffe ou plus rarement injecte, il peut etre a l’origine d’une intoxication severe. Nous rapportons un cas avec hyperthermie maligne et arret cardiaque. Description du cas Un jeune homme de 17 ans aurait consomme du LSD lors d’une soiree chez des amis. Apres des episodes d’hallucinations, il a ete retrouve inanime quelques heures plus tard. A l’arrivee du SAMU, il est en asystolie, avec reprise de l’activite cardiaque apres 3 mn de massage. Le patient est en mydriase bilaterale areactive, il est tachycarde a 126 bpm, febrile a 40,6 °C et convulse. Il est intube, ventile, transfere en reanimation. A l’admission, le score IGSII est de 60 (gravite forte). Il est sedate et refroidi. L’ECG inscrit une tachycardie sinusale a 134 bpm avec des troubles de la repolarisation. Sur le plan biologique on note une acidose metabolique (pH 7,29, bicarbonates 20,5 mmol/L, lactate 2,7 mmol/L), une rhabdomyolyse (CPK 12 762 UI/L), une insuffisance renale aigue (creatinine 144 μmol/L), une insuffisance hepatocellulaire (TP 68 %, TGO 3425 UI/L, TGP 1943 UI/L). Il est hydrate, alcalinise, il n’y a pas necessite de vasopresseurs ni d’hemodialyse. Le patient est extube au 8e jour. Il est agite, agressif, aphasique, apraxique et agnosique. L’IRM cerebrale retrouve des lesions post-anoxiques thalamiques, fronto-parieto-temporales et occipitales. La negativite des analyses toxicologiques, dont le LSD, a conduit a elargir la recherche aux nouvelles drogues de synthese (NDS) : le 25I-NBOMe est dose dans le sang a une concentration de 6,1 ng/mL (LC-MS/MS). L’evolution sur le plan neurologique n’est pas favorable, le patient presente des sequelles : 3 mois plus tard, il a un comportement infantile, reste desoriente dans le temps et dans l’espace, il persiste des troubles attentionnels et executifs, des troubles de l’expression et de la comprehension. Discussion Le 25I-NBOMe fait parti de la famille des NBOMes (NDS) derives des phenethylamines. Ces composes ont une action principale sur les recepteurs serotoninergiques avec une selectivite envers les recepteurs S-HT2A responsables d’effets hallucinogenes. Il n’existe pas d’etude chez l’homme et l’animal, les donnees d’exposition sont celles rapportees dans la litterature medicale [1] . Le nombre d’intoxications est sous-estime, souvent par manque d’identification, seuls quelques laboratoires possedant la technique chromatographique. L’hyperthermie maligne est peu decrite [2] , [3] . Dans notre cas, elle pourrait etre a l’origine de l’arret cardiaque, mais il n’est pas exclu que l’arret ait fait suite a un etat de mal convulsif, le patient n’ayant pas ete vu pendant quelques heures. Au regard de sa toxicite, le 25I-NBOMe est inscrit en France sur la liste des stupefiants (arrete du 6 novembre 2015). Conclusion Le 25I-NBOMe est une nouvelle drogue hallucinogene emergeante. L’intoxication peut etre grave. Son achat sur Internet rend difficile le controle de sa diffusion, une information aux usagers serait souhaitable. D’autre part, il serait important que les laboratoires puissent disposer des etalons permettant son identification.

This paper analyzes taxpayer contributions to state check-off funds using four years of Kentucky household income tax data. An ordered probit shows that a taxpayer tends to give to more check-off funds as income increases and tax price of contribution decreases. A Heckman selection model shows that the Nature Fund and Child Fund are normal goods, with income elasticities of contribution of 0.20 and 0.14. With values of -0.58 and -0.29, the tax price elasticities of contribution are even greater. If refund amount is included, elasticities become smaller; the refund elasticities of contribution are 0.08 and 0.06, respectively.

In France 25,000 new cases of breast cancer are registered every year. To address this situation a new branch of tissular biopathology has been developed: biology transfer supplied by transfer laboratories. By transfer, we mean the privileged relationship between research and clinical biology, corresponding to human tumour expertise, as well as the use of biomarkers before they are put on the market. These laboratories have three scopes of activity: predictive medicine, using informative invasion and proliferation parameters, which enters the clinical strategy at a decision level to adjust therapy and permit a surveillance of therapeutical response; presumptive medicine, relative to the study of the likelihood of being affected by a hereditary cancer, and which allows high risk groups to be identified; and intervention medicine, which consists of contribution of a functional gene in repairing or improving the cell. These transfer laboratories require state of the art technology. In this context, the biomedical engineer must be trained in the terms used in clinical biology for a better understanding, and this for a better tuned analysis of the biologist's needs. He must be vigilant in respecting norms and safety rules in application of the medical biology Guide to Good Analysis Performance (GGAP) and maintain the quality of the equipment.

Results of conservative treatment with surgery and radiation therapy of 132 non-palpable ductal carcinomas in situ of the breast. Purpose. — Retrsopective analysis of results of treatment of 132 subclinical ductal carcinomas in situ, non-palpable. Material and methods. — Patients were treated with limited surgery and 70 Gy radiation therapy (70 Gy). Results. — With a median follow-up of 7 years, the total recurrence rate was 6%, and the actuarial rate at 5 years 4% and at 10 years 13% at. These have no influence on recurrence on the specific actuarial survival rate which was 100% at 10 years. In spite of five infiltrating recurrences of seven, no metastasis appeared 48 months after the salvage surgery. The global rate of breast preservation was 92% at 7 years. Discussion and conclusion. — Therapeutic indications were developed taking into account the present analysis and a literature review (2,338 in situ ductal carcinomas, palpable or not, treated with conservative surgery, with or without adjuvant radio-therapy).

Resume A cote des recepteurs des ˦strogenes et de la progesterone, de nombreux parametres biologiques tissulaires sont proposes dans le cancer du sein comme indicateurs de pronostic ou de reponse therapeutique. Il existe cependant a l'heure actuelle une dissociation entre l'evaluation parfois incomplete de ces facteurs et leur mise a disposition sur le marche, entrainant des couts non controles, voire des consequences nefastes pour les malades. La validation des parametres biologiques doit se concevoir en trois etapes: validation technique (evaluation des diverses approches methodologiques, standardisation des approches choisies et etablissement d'un controle de qualite), validation clinique dans le cadre de protocoles therapeutiques definis et selection des facteurs les plus performants. Ces etapes ne peuvent etre realisees a l'echelon individuel, du fait de l'importance de l'equipement technique et du recrutement necessaires. Le Groupe de biopathologie tissulaire et moleculaire s'est constitue en reseau de 30 laboratoires experts, universitaires, hospitaliers et de centres de lutte contre le cancer. Son but est de contribuer a la mise en place au niveau national de cette strategie d'evaluation, pour permettre la selection de nouveaux facteurs, reellement informatifs et non redondants, et susceptibles d'etre utilises en pratique clinique courante dans le respect des bonnes pratiques de laboratoire.

Objective To ascertain the influence of hormone replacement therapy on clinical and biologic prognostic factors of breast cancer. Methods Between 1976–1992, we treated 1081 postmenopausal women for breast cancer at our institution. Of these, 68 were undergoing postmenopausal hormone replacement therapy at the time of diagnosis. These patients were compared with a matched control group of 272 breast cancer patients who had not undergone prior hormone replacement therapy. Results Patients who developed breast cancer during hormone replacement therapy had fewer locally advanced cancers (large tumors and extensive lymph node involvement) and more well-differentiated cancers (infiltrating lobular cancers and grade 1 cancer). The number of patients with estradiol or progesterone receptors was lower in the hormone-treated group. Metastasis-free survival curves showed a tendency ( P = .05) for better prognosis in hormone-treated patients both overall and in stage T2. Conclusions Hormone replacement therapy per se does not affect the prognosis of breast cancer. Regular surveillance during hormone replacement therapy reduces the number of locally advanced cancers and thus improves the survival rate. The higher number of well-differentiated cancers and the distribution of hormone receptivity may reflect interaction between neoplastic tissue and exogenous hormones.

This study was carried out with three aims: to assess the variations of receptors for beta-oestradiol and progesterone, and for epidermal growth factor (EGF), alpha-fibroblast growth factor (FGF) and beta-FGF, during a spontaneous cycle in normo-ovulatory patients, 2 days before, and 2 and 7 days after the day of ovulation; to compare the control group of patients with those given ovulation induction using a multiparametric approach; and to study the role of luteal supplementation with progesterone in two groups of patients treated by a luteinizing hormone-releasing hormone agonist plus human menopausal gonadotrophin. The results were assessed by biochemical analyses and by immunohistochemistry. In spontaneous cycles, a decrease in steroid receptors occurred during the luteal phase. There was an increase in EGF and EGF receptors which was induced by oestradiol secretion, and an increase during the luteal phase of alpha-FGF and beta-FGF, which are considered to be angiogenic factors. Comparisons between spontaneous and induced cycles showed a few variations in the endometrium but no differences were related to the type of ovulation induction.

Biobanks in general, and specifically tumour banks, are considered as essential tools for the development of translational and clinical research in biology and oncology. Biobank tasks include the collection and preservation of biological samples, and their association with information that will be essential for further scientific use ("annotations" that allow for the "qualification" of biological samples in biological resource). A collection is made of a series of biological resource that are representative of a homogeneous group of individuals or patients that are defined on the basis of clinical or biological information. Collections are used by scientists that are aware of their existence. In the absence of a published catalogue, this awareness is most often limited to research teams that are geographically close, or to investigators who already established collaborative projects with medical teams within the hospital that operates the tumour bank. Publications of catalogues, especially digitalized and online catalogues, should foster the development of high-level, large-scale and multicentric scientific projects. In addition, tumour banks will formalize rules that allow publication of collections, and upstream, rules that are used to qualify biological samples in biological resource: this should translate in an improved overall quality of samples and annotations. Tumour bank catalogues remain relatively few; however, some recent achievements established the "proof of concept" and already raise questions regarding rules for publication. It will be important to demonstrate that these high expectations translate into measurable benefits.

Objectif Poser les bases d’une proposition de consensus pour la realisation d’une expertise medico-judiciaire dans un contexte de deces dans le cadre de la loi Leonetti de 2005. Methodes Monsieur P., âge de 75 ans, a ete decouvert au fond d’un puisard. Extrait par les pompiers, il restait inconscient et etait pris en charge en reanimation ou il devait finalement deceder des suites d’une anoxie cerebrale. L’examen medico-legal du corps (pesant 48 kg pour 172 cm, cachectique) realise a 4 semaines de la prise en charge initiale a conclu que l’interesse a probablement presente un malaise au fond du puisard, possiblement en rapport avec une hypoxie ou la presence de vapeurs toxiques. Le parquet a demande une expertise toxicologique. Resultats Au decours de l’autopsie, du sang cardiaque, du sang peripherique, des urines, de la bile, du contenu gastrique et des cheveux ont ete preleves. Dans le cadre d’une expertise toxicologique de reference, les resultats suivants ont ete obtenus dans le sang peripherique : alcoolemie nulle, HbCO a 1,4 %, cyanures inferieurs a 80 ng/mL, morphine libre a 261 ng/mL, morphine totale a 284 ng/mL et midazolam a 895 ng/mL. L’analyse des cheveux (couleur poivre et sel) sur 3 × 1 cm n’a pas retrouve de morphine (LOD a 20 pg/mg), ce qui confirme que le sujet n’avait pas de traitement a cet analgesique. Le rapport morphine libre/morphine totale est a 0,92, impliquant une administration de morphine peu de temps avant le deces. Les concentrations sanguines de midazolam et de morphine suffisent a expliquer le deces chez un sujet naif et l’expert s’est alors interroge sur la mise en œuvre de soins palliatifs dans le cadre de la loi du 22 avril 2005, dite loi Leonetti relative aux droits des malades et a la fin de vie. Conclusion La loi Leonetti vise a limiter l’acharnement therapeutique. Elle repose sur une ethique fondee sur l’acceptation de la mort et exprimee par la formule « Laisser mourir sans faire mourir ». Elle confirme et codifie des pratiques existantes, comme l’absence d’obstination deraisonnable ou le droit au refus de soins, mais exclut toutefois explicitement le suicide assiste et l’interruption du processus vital des personnes en fin de vie. Dans ce dossier, la morphine et le midazolam, en quantite largement supra-therapeutique, ont directement contribue au deces du patient. Des lors que s’annonce une evolution de la legislation sur la fin de vie suite a la proposition de loi des deputes Leonetti (UMP) et Claeys (PS), il serait important que la communaute des toxicologues etablisse un consensus sur les investigations a pratiquer (quelles molecules, sur quelles matrices) afin de confirmer que le cadre legal est bien respecte.

resume Le controle de qualite europeen de l'EORTC qui s'appuie sur l'analyse de preparations cytosoliques ne cor role pas tous le aspects des dosages biochimiques des recepteurs de l'œstradiol (ER) et de la progesterone (PR). L'analyse aes distribuion, obtenues en routine est, de ce fait, egalement indispensable pour assurer la stabilite des mesures dans le temps, independamment du laboratoire et de la technique employee. Cette etude porte sur 83 907 dosages de recepteurs effectues au cours des 17 dern e es annee, par 17 laboratoires affilies au Groupe francais de biopathologie tissulaire et moleculaire. Les distributions en fonction de l'age montrent une faible variabilite interlaboratoire. Une bonne stabilite dans le temps est demontree pour la technique de liaison d'un radiol gand pou ER et PR, et pour la technique immunoenzymatique (EIA) pour PR. En revanche, on observe pour ER-EIA une augmentation sensible des medane; entre 1987 et 1989.

Both DNA ploidy and S-phase ploidy are promising prognostic factors for node-negative breast cancer patients. Based largely on the analysis of one large study, much of the reported problems with these factors have been caused by some unappreciated complexities in categorizing DNA ploidy into low- and high-risk groups and the lack of some necessary adjustments to eliminate unwanted correlations between DNA S-phase and ploidy. When both DNA ploidy and S-phase are compensated properly, they become independent prognostic markers, forming a powerful prognostic model.

Objectives : The study was carried out on the site of Koussin and Lele in the southern Benin, exploring the efficiency of the supply of the pearl and super granule urea (46%) and NPK 15-15-15 for rice production in the inland valley. Specifically, the study aims to assess the effect of the different forms of urea and NPK 15- 15-15 on the growth and yield parameters of two inland rice varieties (IR841 and NERICA-L14) widely adopted in cultivated lowland of Benin. Methodology and results : The experimental design is a split-plot with two rice varieties, four forms of fertilizer (pearl urea + NPK; pearl urea + super granule NPK; super granule urea + pearl NPK and super granule urea + super granule NPK) in four replications. The growth and yield parameters were assessed. Super granule urea + super granule NPK and super granule urea + pearl NPK have significantly (P < 0.0001) induced higher plant height, numbers of tiller and rice grain per panicle compared with the other treatments whatever the rice variety. However, mean yields of paddy rice and straw were significantly higher (P < 0,0001) for super granule urea + super granule NPK (paddy rice yield : 5,52 ± 0,26 t/ha ; straw yield : 12,19 ± 0,54 t/ha) and super granule urea + pearl NPK (paddy rice yield: 4,82 ± 0,22 t/ha ; straw yield : 10,79 ± 0,61 t/ha). In general with these treatments, the highest yields were recorded with IR841 rice variety. Conclusion and application : The super granule fertilizers are recommended for improving rice production in the inland valley as they improve substantially the yields. In order to improve the efficiency of these fertilizers especially the sources of nitrogen, the study suggests to supply urea super granule + NPK super granule or urea super granule + pearl NPK to improve growth and yields parameter of IR841 rice variety. Keywords : inland valley; super granule fertilizer; soil fertility; irrigated rice production; Benin.

The lack of a standardized methodology for quantifying DNA ploidy and S-phase fraction (SPF) by flow cytometry is hindering routine use of these markers in breast cancer management. In a retrospective clinical multicenter study, we validated a standardized flow cytometry protocol. We tested 633 frozen T(1)T(2), N(0)N(1), M(0) breast tumors obtained in four institutions. Cell preparation was standardized, and precise rules for data interpretation were followed. Three SPF classes were defined on the basis of tertiles after adjustment for ploidy. DNA aneuploidy was observed in 61.0% of cases. No significant difference was observed among centers. Aneuploidy and high SPF were associated with large tumor size, node involvement, high histological grade, and hormone receptor negativity. In the overall population (median follow-up, 69 months), patients with medium and high SPF values had shorter disease-free survival (DFS) than those with low SPF values (P0.0001). Ploidy had no significant influence. By Cox analysis, SPF, pN, and estrogen receptor status were independent predictors of DFS (P = 0.0002, P = 0.001, and P = 0.05). In node-negative patients, SPF was the only predictor of DFS (P = 0.01), whereas in node-positive patients, the risk of relapse increased with both high SPF (P = 0.003) and estrogen receptor negativity (P = 0.004). Low SPF values distinguished grade II tumors with a particularly good outcome. Our results strongly support the use of SPF in multicenter studies and clinical trials and suggest that node-negative patients with slowly proliferating tumors do not require systemic adjuvant therapy.

Thymidylate synthase (TS) is a crucial target for 5-fluorouracil (5-FU) in the de novo pathway of pyrimidine synthesis, which is necessary for DNA synthesis. Thymidine kinase (TK) plays a key role in the complementary or alternative salvage pathway of pyrimidine synthesis in acute or pathological tissue stress. In the present study, the activity levels of TS and TK were determined in 257 primary breast tumors of patients who received tamoxifen as first-line systemic therapy after diagnosis of advanced disease. In 155 (60%) responding patients, the median response duration was 23 months for tumors with low TK activity, 15 months for tumors with intermediate TK activity, and 13 months for tumors with high TK activity (P = 0.003). In Cox multivariate analysis corrected for classical predictive factors including estrogen receptor and progesterone receptor, patients with intermediate and high levels of TK activity in their tumors showed a rapid disease progression (P = 0.0002) and an early death (P = 0.002) after start of tamoxifen treatment. Tumor TS activity levels were not significantly associated with the efficacy of tamoxifen treatment. In 121 patients who became resistant to tamoxifen or additional endocrine treatments and who received 5-FU-containing polychemotherapy, tumor TK activity was not significantly related to the efficacy of chemotherapy. Of the 13 patients with low tumor TS activity, only 1 (8%) responded favorably, whereas 46% (43 of 93) of those with intermediate and 73% (11 of 15) of those with high TS activity responded (P = 0.001). In Cox multivariate regression analysis in which TS was the only significant variable, intermediate and high TS activities were associated with a slow disease progression (P = 0.005) and prolonged survival (P = 0.016) on chemotherapy. In conclusion, for patients with recurrent breast cancer, high tumor TK activity is a significant marker of poor clinical outcome on tamoxifen therapy. Elevated tumor TS activity predicts a favorable outcome for 5-FU-containing polychemotherapy when applied after tumor progression on endocrine therapy.

S-phase fraction (SPF) is a reference for cell-kinetic analysis. In this study, the links between SPF and the essential enzymes participating in the pyrimidine synthesis were investigated in breast cancer and their relationships with the natural history of the disease were compared. We measured thymidine kinase (TK) for salvage synthesis, thymidylate synthase (TS) for de novo synthesis and thymidylate kinase (TMK), which is required for both pathways. Our study population consisted of 211 premenopausal women with node-negative tumors. SPF was assessed prospectively by flow cytometry, whereas enzyme activities were measured retrospectively in cytosols using radioenzymatic methods. Among the enzymes analyzed, only TK demonstrated a strong correlation with SPF (r(s) = 0.59). In univariate analysis, high SPF and high levels of TK were associated with increased risk of developing distant recurrences (p0.001). Correlations with other prognostic factors (histological grade, steroid receptors, DNA ploidy status, urokinase plasminogen activator and plasminogen activator inhibitor type 1) confirmed a parallel association of SPF and TK with the most aggressive tumors. In contrast, TS and TMK were not associated with prognosis. After adjustment for SPF, the risk of relapse increased significantly with TK values. Subgroup analysis showed that additional information was provided by TK in the tumors with low SPF. When urokinase plasminogen activator (uPA) was a candidate variable in multivariate analysis, TK remained significant. Combined with SPF and uPA, TK could be useful to define premenopausal node-negative patients with rapidly proliferating tumors at a high risk of metastatic disease.

High thymidine kinase (TK) activity in cancer cells could counteract adjuvant chemotherapy directed at the inhibition of de novo DNA synthesis. TK is an independent prognostic factor in breast cancer patients receiving adjuvant chemotherapy.In this paper, we describe the effects of extraction and dilution buffer composition on TK enzymatic activity values obtained in breast cancer cytosols with the Prolifigen serum TK-REA kit (Sangtec Medical, Sweden).The addition of MgCl2 and ATP early in the assay, preferably during the extraction of tumor tissue, seems critical to stabilise the enzyme. Furthermore, the use of normal calf serum to dilute both standards and samples is necessary to obtain satisfactory parallelism between TK values in serial dilutions of breast cancer cytosols.Based on the data reported here, the manufacturer has changed the cytosol diluent composition and is adding a specific cytosol assay insert to the Prolifigen TK-REA kit. As evidenced by the laboratory reproducibility, these modifications to the serum assay led to an adequate, standardized protocol for analyzing TK activity in breast tumor cytosols.

As part of a clinical research project, proliferative parameters were studied in primary breast cancer: standardization and technical validation of thymidine kinase (TK), thymidylate synthase (TS) and protein tyrosine kinase (PTK) are described. A total of 633 frozen tumor specimens, available in four institutions, was analyzed in three flow cytometry laboratories for DNA content and percentage of S-phase cells (%S) measurement. 1) The standardization step consisted in developing a common protocol for sample preparation; then, common cell suspensions were analyzed in order to perform an inter-laboratory control. Objective guidelines were elaborated to interpret DNA histograms in breast carcinoma. 2) DNA-aneuploidy was observed in 61% of cases of the retrospective series. Compared with DNA-aneuploid tumors, mean %S was significantly lower in case of DNA-diploidy (respectively: 6.4% and 2.2%, p0.001). When compared between the four institutions, %S distributions did not differ significantly. 3) %S is strongly correlated with TK, TS and PTK and high percentages were also observed in high grade tumors or tumor without hormone receptors. These results show that a standardization in using flow cytometers and DNA software allows multicenter studies.

Retrospective analysis of results of treatment of 132 subclinical ductal carcinomas in situ, non-palpable.Patients were treated with limited surgery and 70 Gy radiation therapy (70 Gy).With a median follow-up of 7 years, the total recurrence rate was 6%, and the actuarial rate at 5 years 4% and at 10 years 13% at. These have no influence on recurrence on the specific actuarial survival rate which was 100% at 10 years. In spite of five infiltrating recurrences of seven, no metastasis appeared 48 months after the salvage surgery. The global rate of breast preservation was 92% at 7 years.Therapeutic indications were developed taking into account the present analysis and a literature review (2,338 in situ ductal carcinomas, palpable or not, treated with conservative surgery, with or without adjuvant radio-therapy).

Thymidine kinase (TK) and thymidylate synthase (TS) play a key role in, respectively, the salvage and the de novo DNA synthesis pathways. TS is a crucial target for 5-fluorouracil(5-FU) and may also influence methotrexate(MTX) efficiency. Tyrosine kinase(TPK) has been associated with the cytoplasmic domain of growth factor receptors as well as oncoproteins. We investigated whether TK, TS and TPK are predictive factors for drug sensitivity evaluated in terms of relapse-free improvement in breast-cancer patients receiving adjuvant chemotherapy. TK, TS and TPK activities were determined in the cytosols of 154 node-positive primary breast cancers. All patients received 5-FU containing adjuvant chemotherapy. Measurements were performed using radioenzymatic methods. The levels of TK were correlated with those of TS and TPK. The levels of TS and TPK were less strongly correlated with each other. High TK levels were more often found in larger tumours, and the levels of both TK and TPK were negatively correlated with those of PgR. Patients whose tumours contained high levels of TK had increased risks of relapse and death. TS was not of prognostic value, while a high level of TPK was associated with early death. In Cox analysis, TK and TPK retained their independent prognostic value. While target enzyme activities on the de novo DNA synthesis pathway could determine response to anti-metabolics mainly inhibiting this pathway, high activities on the alternative salvage pathway could circumvent induced growth inhibition.

All French laboratories that routinely assay estradiol (ER) and progesterone (PR) receptors participate in the European EORTC quality control program based on twice-yearly analysis of 5 cytosolic preparations. This system has considerably reduced inter-laboratory variations, but does not cover all aspects of these assays. Analysis of receptor value distributions is also crucial to ensure that receptor measurements remain stable with time, independently of the laboratory and assay method. This study involved 83907 receptor assays carried out in the last 17 years by 17 laboratories belonging to the French Study Group on Tissue and Molecular Biopathology. The assays were based on radioligand binding (RLA) or immunoenzymology (EIA). For each laboratory, the medians and positivity rates were analysed according to two totally objective criteria, the patient's age and the year of assay, and according to histological grade and histological type of the tumor in order to verify the correlations classically described. Age-related distributions varied little between laboratories, compared with data published by 7 European EORTC laboratories [1]. The results remained relatively stable with time in the RLA method for ER and PR, and in the EIA method for PR. Median ER-EIA data showed a marked increase between 1987 and 1989, mainly due to changes in the quality of Abbott reagents during this period. Otherwise, this analysis confirms previous pathophysiological observations.

The urokinase type plasminogen activator (u-PA) and the plasminogen activator inhibitor-1 (PAI-1) are among the best second-generation prognostic tissue factors in breast cancer. However, different extraction procedures and assay kits are used in different laboratories. A total of 79 breast tumour tissues stored in liquid nitrogen were analysed in this study. We compared u-PA and PAI-1 levels determined with the American Diagnostics (AD) kit after various extraction procedures. The median cytosolic extraction yield in the presence of 0.4 mol/l KCl, calculated relative to extraction in the presence of 10 ml/l Triton X100 when adapted to standard laboratory working hours (incubation for 2 h instead of 12 h) was 74.4% for u-PA and 85.8% for PAI-1. In addition, the correlations were acceptable. Cytosolic extracts prepared with KCl could permit optimal u-PA and PAI-1 assays while also enabling hormone receptors to be determined with the same specimens. Further studies with clinical data are now necessary to determine the prognostic relevance of this extraction procedure.

Health visitors and lay people have been working together in Maidenhead to support women suffering from postnatal illness. Alison Jones, Tricia Watts and Sybil Romain describe how the group supplements medical and psychiatric services, by tackling the isolation women with postnatal depression can feel, and helping them through until the illness ends.

Seven laboratories of the EORTC Receptor Study Group reported the distribution of oestrogen (ER) and progesterone receptors (PR) routinely assayed in breast cancer cytosols. A low interlaboratory variability was demonstrated for the median values, and for the frequency of positive tumours as measured by enzyme immunoassay (EIA). Larger variations were found for the frequency of positive tumours, as measured by radioligand binding assay (RLA). They are probably due to differences in the cut-off levels and in the sensitivity of the assay. Analysis of the variability over time clearly demonstrated that the ER-EIA values initially increased compared with RLA. A possible source of variations could be the calibration drift in the ER-EIA kit. In conclusion, quality assessment of steroid receptors should be monitored by comparison of both common standards and distributions routinely obtained in each laboratory. In-house analysis over time is also essential for reagent survey.

A new method (enzyme-ligand immunoassay, ELIA) is described for the estimation of estrogen (ER) and progesterone (PR) receptors in microsamples of human breast cancer tissue. The technique, based on the nonisotopic measurement of receptor-bound estradiol and progesterone, involves three steps: (a) simultaneous saturation of active receptors with their respective authentic ligands, (b) heat treatment of the cytosol to release the steroids from their cognate receptors before or after absorption with dextran-coated charcoal, and (c) measurement of both steroids present in the cytosol by a modified competitive-inhibition enzyme immunoassay. The useful range of the method was 10-4000 pmol/L for ER and 6.5-1000 pmol/L for PR. The correlation coefficient (r) between the one-point and Scatchard plot analysis was 0.95 for ER and 0.99 for PR. Comparison of the one-point ELIA and expected values with the radioligand binding assay (RLBA) results for EORTC samples gave r = 0.88 and 0.99 for ER and PR, respectively. Further comparison of the one-point ELIA with RLBA and with a commercial enzyme immunoassay, in blind testing of cancer tissue microsamples from 70 patients, gave good agreement for ER with r = 0.95-0.97 and concordance of 92.9-94.4% (cutoff, 15 pmol/g protein) against the other two methods. The results were more disperse in all three methods for PR estimation, the assay correlating perhaps better with the enzyme immunoassay (r = 0.90) at a concordance of 89.4% (same cutoff value).

Pilot retrospective studies have pointed to the prognostic value of thymidine kinase (TK) in breast cancer. We studied the Prolifigen TK-REA assay (Sangtec Medical, Sweden), usually applied to serum, for TK analysis in breast cancer cytosols. Reproducibility was good, provided that small volume pipetting was performed carefully. The TK assay was not influenced by the short-term storage of cytosols in liquid nitrogen or at -80 degrees C. However, some steps appeared critical for good laboratory practice. The TK level was affected by thawing the cytosols more than twice. Tumour storage in liquid nitrogen should be preferred over storage at -80 degrees C. The components of the homogenisation buffer, especially sodium molybdate and KCl can have a marked influence on results. Finally, linearity problems arose with some cytosols. Thus, although assay of TK in cytosols is apparently simple, care must be taken in practice. The TK-REA kit should be standardised before widespread use in breast cancer.

Among the great many prognostic factors currently available in breast cancer, three classes of tissue biological parameters appear to be the most reliable in the establishment of a clinical decision flowchart, when they will have been technically and clinically validated: parameters of hormone dependence, tumour aggressiveness and invasion and parameters of proliferation. This article discusses the difficulties encountered in the evaluation of some of these parameters (hormone receptors by various methodological approaches, proteases, enzymes involved in cell proliferation), with emphasis on standardisation of techniques, development of quality controls, clinical validation and objective information of the medical and scientific communities.

Inflammatory breast carcinoma has to be defined by accurate clinical and pathological criteria. The prognosis is very poor and improvements made by chemotherapy are limited to the increase in the disease-free period and overall survival for a few months.

This study concerns the correlation between ER and PR status, menopausal status and the effect of adjuvant hormonotherapy in high risk primary breast cancer patients. We have compared the results obtained in a randomized trial (Institut Paoli-Calmettes, Marseille) with those of a historic series (Centre René Huguenin, Saint-Cloud). The patients presented the same clinical and histological criteria and received identical therapeutic protocols (chemotherapy and/or hormonotherapy). Compared with patients receiving no adjuvant treatment, it appeared that: 1) in post-menopausal patients, a significant detrimental effect of tamoxifen was found in ER-negative patients, while in ER-positive patients hormonal treatment was well correlated with both the presence and level of steroid receptors; 2) in pre-menopausal patients, hormonal therapy (oophorectomy + tamoxifen) appeared to be mediated by a complex mechanism involving more than an ER-positive cell population. In the light of the published results, the present findings underline the importance of reevaluating the indications of hormonotherapy in terms of hormone receptors and menopausal status. They also indicate the importance of biological factors in the evaluation of response to therapy. They can identify, subsets of patients in whom a given therapeutic protocol is detrimental, even though it may be beneficial for the overall population. heterogeneity in response to therapy among patient subsets is one of the most important problems which confronts medical statisticians and clinical investigators.

To test the qualities of two assays in the same laboratory on the same tumours, a single-point dextran-coated charcoal radioligand binding assay (RLA-DCC) and the Abbott enzyme immunoassay (EIA) were used to perform oestrogen receptor determinations on cytosols from 1841 breast cancers over a 2-year period. Statistical analysis of the data was performed by the Passing-Bablock linear regression technique. The final regression curve between EIA (y) and RLA-DCC (x) yielded y = 1.187 × fmol/mg of protein. However, a high variability in this correlation was observed from 1986 to 1988. This variability could be explained by calibration problems in the immunoassay kits and changes in our technical team. The binding assay appears to be more sensitive to the technicians' experience than the immunoassay. Other technical points are discussed, particularly cytosol preparation and KCl presence or absence in the homogenisation buffer. Finally, the Passing-Bablock and the least squares regression procedures are compared. The conditions allowing optimal correlation and routine determination reliability are defined and the correlation variability is discussed.

In order to test the qualities of the 2 assays, in the same laboratory and on the same tumors, a single-point dextran-coated charcoal radioligand binding assay (RLA-DCC) and Abbott enzyme immunoassay (EIA) were used for more than two years to perform estrogen receptor determinations on cytosols from 2,134 breast cancers. Statistical analysis of the data was performed according to the method of Passing-Bablock. The final regression curve between EIA (y) and RLA-DCC (x) was excellent y = 1.187 X fmol/mg of protein. However, from 1986 to 1988, a great variability was observed for this correlation. We report the study of this variability, which could be explained by several factors, especially calibration problems for the immunoassay kits and changes in our technical team. The binding assay appears to be more sensitive to the technicians' experience than the immunoassay. Technical points are discussed, particularly cytosol preparation and KCl presence or absence in the homogeneisation buffer. The conditions allowing for optimal correlation and routine determination fiability can therefore be defined.

The total amount of cathepsin-D (mature forms and pro-enzyme especially) in the cytosol of 88 breast cancers including 85 primary cancers and 11 axillary lymph node metastasis was measured by way of immuno-radiometric assay. Maximum follow-up is 58 months. Cathepsin-D was found to be independent of clinical and biological parameters including axillary lymph node involvement and oestradiol receptors (RE). Univariate analysis has pointed out a significant linkage between overall survival and cathepsin-D, using a cut-off level of 30 pmol/mg protein which is the most discriminating value. Cathepsin-D appeared to be particularly useful in lymph-node-positive and RE-negative patients but it was not significant in the node-negative population. Multivariate analysis of the overall survival (Cox model) revealed that it was strongly related to oestradiol receptor, lymph node involvement and cathepsin-D.

Urokinase-type plasminogen activator (uPA) and PAIl, are involved in invasive phenotype of several tumors, and have been recently described in malignant gliomas. Expression and tissular localization of PAI2, as well as the clinical relevance of these proteases, need however to be investigated. In the present study, 42 patients with glioma were analyzed for expression of uPA, PAIl, and PAI2 (oligodendrocytoma (n = 2), gliomagrade I–II (n = 6), grade III (n = 6). and grade IV (n = 28). (1) Median cytosolic levels of uPA and PA11 as detennined by EUSA, were respectively 0.03 ng/mg protein (range 0.003–0.16) and 11.9 ng/mg protein (range 0.25–161.8). The highest levels of PAIl were found in grade IV tumors as compared to grades I–III ( P in situ hybridization which localized PAI1 predominantly around neoangiogenic foci, both in tumor and endothelial cells. (2) Expression of PAI2 antigen was heterogeneously distributed among tumors (median = 0.18 ng/mg protein, range 0.02–6.8) but was undetectable in control tissues. This data was confirmed by in situ hybridization. (3) Univariate analysis demonstrated that high levels of PAIl are associated with a shorter disease-free survival both for the overall population ( P = 0.02), and the grades IV ( P = 0.06). In grade IV gliomas, high levels of PAI2 are, in contrast, highly correlated to a better overall survival rate at 18 months (48% vs 0%, P = 0.015). Our preliminary results suggest that, in malignant gliomas, PAIl and PAI2 may be useful in the analysis of therapeutic protocols. Further studies should precise their biological role, in order to evaluate them as potential therapeutic target.

We analyzed erbB -2 gene amplification in 170 primary breast carcinomas. Thirty-one percent of tumors exhibited additional copies of erbB -2 gene. Chi-square analysis did not elicit any association between gene amplification and either menopausal or node status. A slight trend was observed with respect to the SBR grading. In contrast, significant correlation was associated with the age of patients and we found a strong relation with the intratumoral steroid receptor status. erbB -2 amplification significantly occurs in tumors whose estrogen receptor and progesterone receptor were below the cut-off value or absent and tumors with dissociated estrogen receptor and progesterone receptor status were revealed as entities similar to both estrogen receptor and progesterone receptor negative tumors. erbB -2 gene/hormonal receptors/breast cancer