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Background: Breast cancer (BC) is a heterogeneous disease and can be classified according to the expression of four genes: estrogen receptor (ER), progesterone receptor (PGR), human epidermal growth factor receptor 2 (HER2) and marker of proliferation KI67. Four groups of BC have been described: Luminal A (ER+ or PGR+, KI67- HER2-), Luminal B (ER+ or PGR+, KI67+ or HER2+), HER2 (ER- HER2+) and triple negative (ER- PGR- HER2-). Autophagy is a lysosomal degradation pathway which plays a crucial dual role in tumorigenesis, producing pro-survival or pro-death activity. Altered autophagy has been observed in BC but no study has described transcript level variations of "autophagy genes" according to sub-groups. In order to further explore those alterations, we analyzed gene expression of 40 "autophagy genes" in normal and tumor cells using public transcriptomic data. Samples and Methods: 5497 transcriptomes were obtained from raw data downloaded from public databases. Two distinct Affymetrix series were built after GC-RMA normalization: one from HG-U133A arrays (n=2806) and one from HG-U133 plus2 arrays (n=2691). Each series was standardized with the Aroma R package that was designed to normalize multicenter extremely large Affymetrix data sets. The same computations were performed in the two data set in order to cross-validate the results. Samples were classified with a reduced number of genes (such as for St Gallen classification) and with classifiers using 50 or more than 300 genes (PAM50, CIT and IntClust centroids). Results: In each of the two series, there were about 500 Basal, 300 HER2, 1800 Luminal tumors and 100 normal epithelial breast cells. Positivity or negativity of four genes (ER, PGR, KI67, HER2) produced robust classifications concordant with PAM50 classification based upon centroids. Within the 40 "autophagy genes" studied, at least 10 genes were significantly correlated with one or two sub-groups: ATG3 and ATG9A were associated with ER- tumors including basal group; ATG2B , BECN1 and ULK2 were positively correlated with ER expression and luminal subtypes; ULK1 and RAB24 were associated with ER+HER2+ tumors and MAP1LC3B and ATG5 with ER-HER2+ subgroup; GABARAPL1 was more expressed in normal breast tissue than in cancer cells. Discussion: Exploring autophagy in large-scale transcriptome data, we confirmed a previous result showing that GABARAPL1 expression is reduced in breast cancer cells compared with the normal epithelial cells. Interestingly, transcript levels of "autophagy genes"were not evenly distributed among the different tumor subtypes. For instance, some genes were preferentially expressed in ER+ subtypes and others in ER- tumors, suggesting that autophagy might play different roles in the different subgroups, providing different potential targets for therapy. Citation Format: Feugeas J-P, Belmiloudi S, Boyer-Guittaut M, Peixoto P, Hervouet E. Relationships between breast cancer subtypes and expression of autophagy related genes [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-06-09.

Les puces a ADN en cancerologie sont des outils d’une nouvelle discipline de recherche, l’oncogenomique. Les puces dites « pangenomiques » detectent a l’echelle du genome entier les lesions de l’ADN (amplifications, deletions et autres mutations), permettent de preciser les interactions proteines-ADN dans les regions promotrices (puces ChIP-on-chip) et quantifient les niveaux d’ARN (puces d’expression, puces pour microRNA). L’analyse des donnees generees necessite des moyens statistiques specifiques aux grandes bases de donnees et est devenue une discipline a part entiere de la bioinformatique. L’objectif de ces analyses est de mettre en evidence de nouvelles cibles therapeutiques et de fournir un nombre limite de marqueurs biologiques pertinents pour la classification ou le pronostic des cancers. Ces marqueurs sont accessibles aux techniques de routine des laboratoires de biologie clinique (PCR, immundosages...) ou d’anatomie pathologie (immunohistochimie). Neanmoins la reduction des couts de production des puces rend possible aujourd’hui l’utilisation de puces dediees dans les laboratoires d’analyses pour detecter des signatures moleculaires basees sur 50 a 100 genes ou pour la detection rapide de mutations de suppresseurs de tumeurs comme p53.

Resume Objectifs Nous avons compare le dosage semi-quantitatif d’acetoacetate par bandelette urinaire avec le dosage capillaire quantitatif du principal corps cetonique, le 3-beta-hydroxybutyrate (3-HB), pour le diagnostic d’acidocetose aux urgences. Methodes Etude retrospective sur un an des patients qui presentaient lors du triage une hyperglycemie ≥ 250 mg/l. Nous avons compare sensibilite, specificite, valeurs predictives (VP) et rapports de vraisemblance (RV) de la cetonurie et de la cetonemie pour le diagnostic d’acidocetose (cetonurie ou cetonemie, bicarbonates sanguins 16 meq/l) sur l’echantillon de patients pour lesquels la cetonurie, la cetonemie et l’ionogramme etaient disponibles. Resultats Nous avons etudie 355 patients hyperglycemiques. La mediane du delai d’obtention de la cetonurie (77 % des malades) etait de 21 min. Ce delai etait superieur a 2 heures dans plus de 10 % des cas. La comparaison entre cetonurie et cetonemie etait possible pour 173 patients (dont 6 % en acidocetose). Une cetonurie egale a une croix ou moins, ou une cetonemie inferieure a 3 mmol/l permettaient d’exclure l’acidocetose (VP negative 100 %). Au seuil de deux croix de cetonurie et de 3 mmol/l de cetonemie, les deux tests avaient la meme sensibilite (100 %), mais la specificite du 3-HB (94 %) etait significativement plus elevee (p Conclusion Le dosage capillaire du 3-HB est plus rapide a obtenir et plus performant que la cetonurie pour detecter une acidocetose a l’accueil des urgences.

Background Breast cancer is currently classified in 3 groups based on estrogen receptor alpha (ER) and human epidermal growth factor receptor 2 (HER2/ERBB2) gene expression: one basal-like (ER-ERBB2-), one HER2−enriched (ERBB2+) and one luminal (ER+). Yet, in transcriptome-based classifications, ER-ERBB2+ group partially overlaps with more recently defined ER-AR+ (androgen receptor positive) group. This type was named molecular apocrine, in reference to the histopathologically characterized apocrine carcinomas (H-Apo), in which a marked activation of AR signaling was demonstrated with a distinct proteomic signature. H-Apo tumors correspond to 1% of invasive breast carcinomas and are clearly morphological distinct from other AR+ tumors. However, no specific H-Apo transcriptome signature has been reported for this sub-group. In an effort to better characterize those tumors, we have performed a meta-analysis of genomic data, focusing on the ER- AR+ breast subset. Samples and Methods: Chips were from Affymetrix array generations HG-U133. 258 profiles were unpublished and 1145 were from published or in press data. Gene expression was carried out after GC-RMA normalization. Unsupervised hierarchical clustering and other statistical analysis were performed with R software. Results: 160 of the 1403 investigated tumors were ER-AR+. An unsupervised hierarchical clustering clearly identified a small subgroup of 14 closely tumors expressing high transcripts levels of PIP, HPGD, ACSM1, AR, SDR5A1, HS3DB1. This profile was very similar to the proteomic signature previously described for the H-Apo tumors. In addition, the pathology report, although available only for 4 of those14 tumors, described them as typical apocrine carcinomas. Taken together, these data suggested that this cluster was the H-Apo subgroup. Unexpectedly, when using the transcriptomic PAM50 classification, 13 were classified as Luminal and only 1 as HER2−enriched, although the 14 tumors were all ER-negative. CGH analysis with Agilent 244K chips was carried out with 25 ER- AR+ tumors, of which 5 were H-Apo carcinomas. Importantly, those 5 H-Apo tumors exhibited fewer DNA lesions than the other ER-AR+ apocrine tumors (17% copy number alterations in H-Apo group versus 41%, p=0.02). More CGH data are currently under investigations and will be discussed. Discussion: The histopathologically characterized apocrine carcinomas (H-Apo) display transcriptomic signs of active androgen metabolism and fewer DNA lesions than others molecular apocrine tumors. This could suggest that molecular apocrine and H-apocrine tumor derive from the same cell of origin, but that only H-Apo retains morphological apocrine features, possibly due to the presence of fewer genetic lesions. In any case, the prominent androgen signaling activation warrants functional assays of anti-androgen in these breast cancer subtypes. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-05-01.

To analyse HLA and insulin-dependent diabetes mellitus (IDDM) association in the ethnically mixed population of La Reunion island, we carried out a family study on 70 diabetic subjects. HLA-DQA1, -DQB1 and -DRB1 typing was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), completed by PCR-sequence-specific oligonucleotide (SSO) and PCR-sequence-specific priming (SSP). Haplotype-relative risks (HRR) were determined with the non-transmitted parental haplotypes as controls, and relative risks (RR) were calculated with a classical case-control study. The most significant risks were found for the cis and trans combinations between DQA1*03 or *0501 (Arg52+) and DQB1*02 or *0302 (Asp57-) alleles, suggesting a direct role for the HLA-DQ heterodimer in IDDM susceptibility. Interestingly, due to the mixed origin of the population, the trans-encoded DQ molecules in the (DR3)-DQA1*0501-DQB1*02/(DR4)-DQA1*03-DQB1*0302 subjects were also found cis-encoded in patients with the (DR7 or 9)-DQA1*03-DQB1*02 haplotype and in a patient with the rare (DR11)-DQA1*0501-DQB1*0302 haplotype. A relative predispositional effect (RPE) analysis gave significant haplotype-IDDM+ associations in the following order: (DR3)-DQA1*0501-DQB1*02 > (DR4)-DQA1*03-DQB1*0302 > (DR9)-DQA1*03- DQB*02 > (DR7)-DQA1*03-DQB1*02 > (DR2)-DQA1*01-DQB1*0502. No protective effect remained significant once the susceptible haplotypes were removed. A stratification study showed a stronger influence of the DQ genes than DRB1 alleles within the DR7 haplotypes. On the other hand, IDDM subjects with only one susceptible haplotype had inherited this haplotype more often from their father than from their mother. This paternal effect could be related to the greater risk of IDDM in offspring of diabetic fathers than the risk in offspring of diabetic mothers.

In advanced breast cancers, TP53 mutation is highly predictive of complete response to high-dose epirubicin/cyclophosphamide chemotherapy. In these tumours with an altered control of genomic stability, accumulation of chemotherapy-induced genetic alterations may contribute to cell death and account for complete response. To explore the effects of chemotherapy on stability of the tumour genome, allelic profiles were obtained from microdissected tumour samples before and after chemotherapy in 29 unresponsive breast cancers (9 with TP53 mutation). Ninety-four per cent allelic profiles remained unchanged after treatment. Interestingly, 11 profiles (6%) showed important changes after treatment; allelic imbalances significantly increased (four cases) or decreased (seven cases) after chemotherapy in three distinct experiments, two of which using laser microdissected tumour cells. These genetic changes were not linked to the TP53 status, but one tumour showed complete disappearance of TP53-mutated cells in the residual tumour after treatment. Altogether, these observations carry important implications for the clonal evolution of breast cancers treated with DNA-damaging agents, as they point both to the importance of tumour heterogeneity and chemotherapy-driven selection of subclones.

In emergency, some "low cost" biological tests are too often systematically performed while others are not prescribed because they are considered "too expensive". Good practices for biological testing are in fact the real means for saving money. Here, in order to help for defining those good practices, we review emergency medical approach and, as an example, specify findings concerning the clinical utility of key blood analyses in patients with acute dyspnea. Emergency laboratory testing is usefull when it contributes to establish the diagnosis or to evaluate comorbidity or to stratify disease severity. In a given emergency context, clinical utility must be anticipated according to a bayesian approach with an estimation of the post-test probabilities using the likelihood ratios (estimated from literature) and the pretest probabilities (established by examination at the bedside). The likelihood ratio is the best criterion for diagnostic accuracy of a biological test. According to this criterion, troponin, natriuretic peptides, procalcitonin and D-dimers are four "costly" markers but which can significantly contribute to the etiologic diagnosis of an acute dyspnea. Troponin, natriuretic peptides and procalcitonin are also prognostic markers and are valuable parameters for stratifying disease severity according to their initial value and their plasmatic kinetic during the clinical course of the disease. In conclusion, it is not only the cost of the test but overall the potential impact of its result on the management of the patient's care which makes the decision of performing the test or not.

Biochemical and pharmacological tests usually prescribed in casualty department were reviewed taking into account their physiological significance and predictive value : ions, total proteins, carbohydrate and nitrogenous metabolites, enzymes, tissue markers, pharmacological drugs. Few blood components were kept with the first intention, ideally with a turn around time below one hour: sodium, potassium, chloride, bicarbonate, total proteins, pCO2 and pO2, creatinine, glucose, ketone compounds, calcium, bilirubin, transaminases, lipase, C-reactive protein, myoglobin, troponin, chorionic gonadotropin hormone. Those tests do not have to be systematically performed but prescribed only after the evaluation of pre-test probabilities by the clinician.

Self peptides bound to HLA-DQ7 (alpha1*0501-beta1*0301), one of the HLA molecules associated with protection against insulin-dependent diabetes mellitus, were characterized after their acid elution from immunoaffinity-purified HLA-DQ7 (alpha1*0501-beta1*0301) molecules. The majority of these self peptides derived from membrane-associated proteins including HLA class I, class II, class II-associated invariant chain peptide and the transferrin-receptor (TfR). By in vitro binding assays, the specificity of these endogenous peptides for HLA-DQ7 (alpha1*0501-beta1*0301) molecules was confirmed. Among these peptides, the binding specificity of the TfR 215-230 self peptide was further examined on a variety of HLA-DQ and DR dimers. Several findings emerged from this analysis: (1) this peptide displayed HLA-DQ allelic specificity, binding only to HLA-DQ7 (alpha1*0501-beta1*0301); (2) when either the DQalpha or DQbeta chain was exchanged, little or no binding was observed, indicating that specificity of HLA-DQ peptide binding was determined by polymorphic residues of both the alpha and beta chains. (3) Unexpectedly, the TfR 215-230 self peptide, eluted from DQ, was promiscuous with regard to HLA-DR binding. This distinct DR and DQ binding pattern could reflect the structure of these two molecules as recently evidenced by crystallography.

The potential role of tumour necrosis factors (TNFs) in autoimmunity and insulin-dependent diabetes mellitus (IDDM) led us to determine in vitro TNF-alpha and lymphotoxin-alpha (LT-alpha, TNF-beta) production in IDDM patients according to TNF polymorphism. LT-alpha production of peripheral blood mononuclear cells (PBMC) was lower in diabetic subjects (m = 0.30 +/- 0.2 ng.10(-6) cells) than controls (m = 0.68 +/- 0.3 ng.10(-6) cells, p0.05), and early age-at-onset was correlated with low LT-alpha production (rs = 0.8, p = 0.0006). TNF-alpha production was the same in patients and controls, but patients with HbA1cor = 8% had a higher TNF-alpha production (m = 3.05 +/- 1.2 ng.10(-6) cells) than those with HbA1c8% (m = 1.31 +/- 0.33 ng.10(-6) cells, p0.05). A study of the microsatellite TNFa region close to the LTA gene showed that the presence of the TNFa1 allele in HLA-(DR3) subjects was associated with increased risk of IDDM. TNFa1-positive subjects (both patients and controls) also had lower LT-alpha production than other subjects. These results indicate that low LT-alpha production is an additional risk factor for IDDM and that poor glycaemic control in patients is associated with enhanced PBMC TNF-alpha production which causes an imbalance between TNF-alpha and LT-alpha production in IDDM patient.

Tumor protein 53 (TP53) is the most commonly altered gene in human cancers. In breast cancers, TP53 is mutated in approximately 30% of all cases, but this frequency fluctuates widely within the different molecular subclasses. Different types of mutations may be observed, such as substitutions (replacement of a nucleotide pair by another one), or complex mutations (deletions or insertions of one or more nucleotides). Mutation types may reflect mechanism of DNA lesion or DNA repair deficiencies. Furthermore, mutations can give rise to different effects such as truncating mutations leading to loss of function, or missense mutations often leading to dominant negative activity. Those mutation effects can be advantageous in tumorigenesis and thus can be subject to selective pressure. Here we classified 572 breast tumors in three groups, according to microarray data: luminal, basal and molecular apocrine. TP53 status was assessed by a yeast-based functional assay (FASAY) and cDNA sequencing. We then assessed whether any feature of TP53 mutations would be preferentially associated to a specific subtype of breast cancer. - In term of TP53 mutation frequencies, as expected, lowest frequency was observed in luminal subgroup (26%) and highest in basal (90%) and molecular apocrine (70%). Notably, much higher rate of TP53 mutations occurred in luminal B subgroup (41%) than in luminal A one (17%), suggesting that TP53 may be an important feature in progression from luminal A to B. - In term of mutations types, luminal tumors showed high frequency of substitutions, while molecular apocrine and basal presented increased rate of deletions and insertions, reflecting probably increased rate of DNA breaks. This suggests that same mutational events may occur in basal and molecular apocrine tumors. - In term of mutation effects, we found high frequency of missense mutations in luminal tumors (notably AT to GC) and much higher rate of truncating mutations in basal tumors. These observations point to an existence of different selection pressure in each of them, such as a strong pressure for P53 mutations with potential dominant negative inhibition of P73/P63 (recently shown to favor invasion), in luminal tumors. Collectively, these results point not only to different mechanisms of P53 gene inactivation, but also different functional consequences among the different breast cancer subclasses. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-02-09.

The possible role of tumor necrosis factors (TNF) in autoimmunity led us to explore the relationship between TNF production, polymorphism of the TNF-beta gene and type one diabetes. In the diabetic group we found a low production of TNF-beta. This abnormality appeared not to be exclusively related to the TNF-beta gene and could be also due to polymorphic regulatory sequences in the major histocompatibility complex region.

Platelet transfusions were introduced in the early sixties. However they have only been developed when plastic bags could be used as blood containers. Another important step was achieved when cell separators have permit to obtain large amount from one blood donor. Platelet transfusions are now a common therapeutical approach of primary thrombocytopenia or cytopenia secondary to chemotherapy. Their development has pointed out two aspects: first the role of platelet alloantigen, second the importance of the technique used to collect platelet and the conditions of platelet storage.

Glucose transport activity was determined on erythrocytes from healthy adult and children. A large variability of values was found between each donor, but transport rate was significantly higher in erythrocytes from adults than those from children. In opposite, the same number of glucose transport sites was found on erythrocytes from adult and children, suggesting that transporters from adult erythrocytes exhibited an higher own activity than transporters from child erythrocytes. Involvement of various factors in glucose transport activity was discussed.