Search

Les infections urinaires occupent le deuxième rang des infections bactériennes de l’enfant. Les entérobactéries sont majoritairement en cause. Depuis la fin des années 1990, l’émergence d’entérobactéries productrices de bêta-lactamase à spectre étendu (EBLSE) est observée dans plusieurs régions du monde et dans diverses situations cliniques. L’objectif principal de notre travail est de répertorier les pyélonéphrites aiguës (PNA) à EBLSE chez les enfants pris en charge en 2014 dans 5 services pédiatriques membres du Réso-Infectio-PACA-Est, et comparer leurs caractéristiques clinico-biologiques avec celles des PNA dues à des entérobactéries non productrices de BLSE (ENBLSE). Nous avons conduit une étude rétrospective multicentrique de type cas-témoin en 2014. Nous avons recensé 12 PNA à EBLSE et 486 PNA à ENBLSE. Parmi ces dernières, 56 ont été tirées au sort et comparées avec les cas EBLSE (4 témoins pour 1 cas). Les caractéristiques cliniques des 2 populations étaient comparables, mais au moins un facteur de risque de développer une infection urinaire à EBLSE était présent dans 100 % des cas du groupe EBLSE (pLes EBLSE restent sensibles dans 78% des cas aux aminosides, qui pourraient être une alternative de choix permettant l’épargne des carbapénèmes. Malgré une adaptation thérapeutique hétérogène, après réception de l’antibiogramme, tous nos cas de PNA à EBLSE ont été considérés comme évoluant favorablement avec épargne de traitement par excès par carbapénèmes (16% des traitements d’adaptation).

Summary Testicular germ cell tumours (TGCTs), the most frequent solid tumour of the young men, originate from the primitive germ cells. They share some pluripotency stem-cell markers which may help to distinguish between seminoma, the most frequent TGCTs and non-seminoma tumours, such as embryonal carcinoma, teratocarcinoma or choriocarcinoma. Due probably to the propensity of seminoma to apoptosis, only two cell lines originated from pure testicular seminoma, TCam-2 and JKT-1 have been up to now, established, maintained and proposed as representative models of human testicular seminoma. However, both seem, following recent reports, to be able to drift. Thus, the molecular signature of embryonic stem-cell markers of the JKT-1 cells cultured in our laboratory, were studied by RT-PCR, Western blot and immunofluorescence (IF). JKT-1 cells analysed after 30 passages, expressed placenta alkaline phosphatase but not alphafoetoprotein (αFP) nor beta-human chorionic gonadotropin. JKT-1 cells also expressed markers of pluripotency such as NANOG and OCT3/4 and more specific seminoma markers, such as AP2γ and HIWI. However, protein expression of OCT3/4 and AP2y was weak and these JKT-1 cells expressed SOX2, a marker of embryonal carcinoma and did not express c-KIT usually expressed in most seminoma. Possible derivation through in vitro culture conditions was supported by looking at later passages (61) which showed a decrease of NANOG and HIWI protein expression. JKT-1 cells express a signature of markers which is still near from the one express by seminoma cells, allowing carcinogenetic studies. However, because of their great ability to drift as shown for TCam-2, it is recommended to verify and to precise this molecular signature before reporting functional results.

Background Fetal exposure to environmental estrogens may contribute to hypofertility and/or to testicular germ cell cancer. However, many of these xenoestrogens have only a weak affinity for the classical estrogen receptors (ERs,) which is 1,000-fold less potent than the affinity of 17β-estradiol (E2). Thus, several mechanisms have been suggested to explain how they could affect male germ cell proliferation at low environmental relevant concentrations. Objectives In this study we aimed to explore the possible promoting effect of bisphenol A (BPA) on human testicular seminoma cells. BPA is a well-recognized estrogenic endocrine disruptor used as a monomer to manufacture poly carbonate plastic and released from resin-lined food or beverage cans or from dental sealants. Methods and results BPA at very low concentrations (10−9 to 10−12 M) similar to those found in human fluids stimulated JKT-1 cell proliferation in vitro. BPA activated both cAMP-dependent protein kinase and cGMP-dependent protein kinase pathways and triggered a rapid (15 min) phosphorylation of the transcription factor cAMP response-element–binding protein (CREB) and the cell cycle regulator retinoblastoma protein (Rb). This nongenomic activation did not involve classical ERs because it could not be reversed by ICI 182780 (an ER antagonist) or reproduced either by E2 or by diethylstilbestrol (a potent synthetic estrogen), which instead triggered a suppressive effect. This activation was reproduced only by E2 coupled to bovine serum albumin (BSA), which is unable to enter the cell. As with E2-BSA, BPA promoted JKT-1 cell proliferation through a G-protein–coupled nonclassical membrane ER (GPCR) involving a Gαs and a Gαi/Gαq subunit, as shown by the reversible effect observed by the corresponding inhibitors NF449 and pertussis toxin. Conclusion This GPCR-mediated nongenomic action represents—in addition to the classical ER-mediated effect—a new basis for evaluating xenoestrogens such as BPA that, at low doses and with a high affinity for this GPCR, could interfere with the developmental programming of fetal germ cell proliferation and/or differentiation when they cross the placenta.

Introduction L’objectif de notre travail est de repertorier les pyelonephrites aigues (PNA) a d’enterobacteries productrices de betalactamase a spectre etendu (EBLSE) chez les enfants pris en charge en 2014 dans 5 services pediatriques membres du reso-infectio-PACA-Est et comparer leurs caracteristiques clinico-biologiques avec celles des PNA dues a des enterobacteries non productrices de BLSE (ENBLSE). Materiels et methodes Nous avons conduit une etude retrospective multicentrique sur l’annee 2014. Nous avons recense 12 PNA a EBLSE et 486 PNA a ENBLSE. Parmi ces dernieres, 56 ont ete tirees au sort et comparees avec les cas EBLSE. Des donnees complementaires permettant d’approcher l’evolution temporelle des PNA a EBLSE chez l’enfant dans le Sud-Est de la France entre 2010 et 2015 ont ete recueillies. Resultats Les caracteristiques cliniques des 2 populations etaient comparables, mais au moins un facteur de risque de developper une infection urinaire a EBLSE etait present dans 100 % des cas du groupe EBLSE ( p 0,001). Une antibiotherapie curative dans les 3 mois precedents est retrouvee chez 41,7 % des cas versus 3,6 % chez les temoins ( p = 0,001). Aucune difference significative n’a ete retrouvee dans les 2 groupes concernant la presentation clinico-biologique. Escherichia coli est majoritaire dans toutes les PNA. Aucune enterobacterie n’etait resistante aux carbapenemes. Les EBLSE restent sensibles dans 78 % des cas aux aminosides. Malgre une adaptation therapeutique heterogene, tous nos cas de PNA a EBLSE ont ete consideres comme evoluant favorablement avec epargne de traitement par exces par carbapenemes (16 % des traitements d’adaptation). Conclusion Notre etude souligne une prevalence non negligeable et stable dans le temps des PNA a EBLSE, avec une surrepresentation dans les centres universitaires et une difficulte de reperage anamnestique, compliquee par l’absence de specificite clinique et biologique de ces infections par rapport aux PNA a ENBLSE. Nous avons mis en evidence l’heterogeneite des pratiques en termes d’antibiotherapie, rapportees a l’absence, en 2014, de consensus national ou de protocoles locaux pour ces infections. Malgre cela, les PNA a EBLSE chez l’enfant dans notre etude ont fait l’objet d’une prise en charge sans exces therapeutique en particulier concernant l’epargne des carbapenemes. Les recommandations pediatriques recentes (SPILF et reseau de pediatrie regional), qui promeuvent l’usage des aminosides en monotherapie, encouragent la diminution de consommation de C3G, l’epargne des carbapenemes et la desescalade antibiotique, et nous apparaissent justifiees.

Testicular germ cell tumours are the most frequent cancer of young men between 15 and 35 years of age. While pathogenesis and reasons of an increasing incidence (about 2% each year) all over the world remain unknown, epidemiological and clinical data have suggested that fetal exposure to environmental endocrine disruptors (EEDs) with estrogenic effects, could participate to testicular germ cell carcinogenesis. However, EEDs (like bisphenol A) are often weak ligands for classical nuclear estrogen receptors. Using a human seminoma cell line (JKT-1), which expressed ER beta but not ER alpha, we previously reported that bisphenol A (BPA) could induce cell proliferation at a very low concentration (10-9M) independently of the classical estrogen receptor ER beta (Bouskine et al., Endocrinology, 2008). The non classical membrane G-protein coupled estrogen receptor (GPER or GPR30) has been recently shown to mediate the effects of several xenoestrogens through rapid non genomic act ivat ion of signal transduction pathways in various human estrogen dependent cancer cells (breast, ovary, endometrium).

Clinical and experimental studies have suggested that estrogens, the archetype of female hormones, participate in the control of male germ cell proliferation and that fetal exposure to environmental estrogens may contribute to hypofertility and/or to testicular germ cell cancer. However, the underlying mechanisms remain to be elucidated. 17β-Estradiol (E2) conjugated to BSA was able to stimulate human testicular seminoma cell proliferation by triggering a rapid, nongenomic, membrane-mediated activation of ERK1/2 and cAMP-dependent protein kinase A (PKA). Both ERK1/2 and PKA participated in this promoting effect. This activation was associated with phosphorylation of the transcription factor cAMP response element-binding protein and the nuclear factor retinoblastoma protein. Enhanced proliferation together with ERK activation could be reversed by pertussis toxin, a G protein inhibitor. Estrogen receptors (ERs) in JKT-1 were characterized by immunofluorescence, subcellular fractioning, and Western blot. JKT-1 cells did not express ERα but ERβ, which localized to the mitochondria and the nucleus but not to the membrane. Moreover, neither ICI-182,780, a classical ER antagonist, nor tamoxifen, a selective ER modulator, could reverse the 17β-estradiol-BSA-induced promoting effect. Estrogens contribute to human testicular germ cell cancer proliferation by rapid activation of ERK1/2 and PKA through a membrane nonclassical ER. This nongenomic effect represents a new basis for understanding the estrogenic control of spermatogenesis and evaluating the role of fetal exposure to xenoestrogens during malignant transformation of testicular germ stem cells.

In a recent issue of Cancer Biology & Therapy, Vitale et al. introduced the paper of Franco et al. reporting the expression of GPER (a seven-transmembrane-spanning receptor also known as GPR30) on testicular germ cell tumors (TGCTs). The paper commented on the possible pathophysiological role of this G-protein-coupled receptor (GPCR) during testicular germ cell carcinogenesis. They also proposed the potential use of selective antagonists of GPER as new-targeted therapy in these tumors. We agree with this hypothesis and would like to highlight here some published results supporting this hypothesis and give some details concerning the role of estrogens and xeno-estrogens in testicular carcinogenesis and the effects of GPER antagonist in human seminoma cells. We are sure that this information we obtained by working for several years on a human seminoma cell line will be appreciated by the readers of Cancer Biology & Therapy.

Dear Sir, Testicular germ cell tumors (TGCT) are the most frequent cancer affecting young men with an increasing incidence all over the world. Pathogenesis and reasons of this increase remain unknown. Epidemiological, clinical and molecular data have suggested that fetal exposure to environmental endocrine disruptors with estrogenic effects could participate in testicular germ cell carcinogenesis by influencing the fate of germ stem cells that share molecular markers with the malignant germ cells. However, no animal model supports this hypothesis, and even estrogen dependency of TGCT has not yet been fully demonstrated. Using the JKT-1 cell line derived from a human testicular seminoma and seminoma tumors, the most frequent TGCTs, we have previously reported that seminoma cells expressed functional aromatase, which is able to convert testosterone into estradiol as well as estrogen receptor beta (ERb) but not ERa. Estradiol 17b (E2) at physiological concentrations was able to suppress in vitro JKT-1 cell proliferation through ERb. However, when E2 was conjugated to bovine serum albumin (E2-BSA), which does not cross the membrane, the effect observed was completely different. E2BSA was able to stimulate JKT-1 cell proliferation by activating the PKA pathway with a rapid (15 min) phosphorylation of the transcription factor cAMP response element binding protein (CREB), through a membrane G-protein-coupled receptor (GPCR). Similar results were obtained with bisphenol A (BPA) at low (pM to nM) concentrations. BPA, a well-recognized estrogenic endocrine disruptor, is used as the monomer to manufacture polycarbonate plastic, and it is released from resin lining of canned foods or beverages or from dental sealants. It is believed to be involved in developmental, reproductive and malignant diseases by mimicking the natural hormone E2 and interfering with

Background Testicular germ cell tumours are the most frequent cancer of young men with an increasing incidence all over the world. Pathogenesis and reasons of this increase remain unknown but epidemiological and clinical data have suggested that fetal exposure to environmental endocrine disruptors (EEDs) with estrogenic effects, could participate to testicular germ cell carcinogenesis. However, these EEDs (like bisphenol A) are often weak ligands for classical nuclear estrogen receptors. Several research groups recently showed that the non classical membrane G-protein coupled estrogen receptor (GPER/GPR30) mediates the effects of estrogens and several xenoestrogens through rapid non genomic activation of signal transduction pathways in various human estrogen dependent cancer cells (breast, ovary, endometrium). The aim of this study was to demonstrate that GPER was overexpressed in testicular tumours and was able to trigger JKT-1 seminoma cell proliferation. Results We report here for the first time a complete morphological and functional characterization of GPER in normal and malignant human testicular germ cells. In normal adult human testes, GPER was expressed by somatic (Sertoli cells) and germ cells (spermatogonia and spermatocytes). GPER was exclusively overexpressed in seminomas, the most frequent testicular germ cell cancer, localized at the cell membrane and triggered a proliferative effect on JKT-1 cells in vitro, which was completely abolished by G15 (a GPER selective antagonist) and by siRNA invalidation. Conclusion These results demonstrate that GPER is expressed by human normal adult testicular germ cells, specifically overexpressed in seminoma tumours and able to trigger seminoma cell proliferation in vitro. It should therefore be considered rather than classical ERs when xeno-estrogens or other endocrine disruptors are assessed in testicular germ cell cancers. It may also represent a prognosis marker and/or a therapeutic target for seminomas.