Genomic characteristics and clinical significance of CD56+ circulating tumor cells in small cell lung cancer.

Circulating tumor cells (CTC) have been studied in various solid tumors but clinical utility of CTC in small cell lung cancer (SCLC) remains unclear. The aim of the CTC-CPC study was to develop an EpCAM-independent CTC isolation method allowing isolation of a broader range of living CTC from SCLC and decipher their genomic and biological characteristics. CTC-CPC is a monocentric prospective non-interventional study including treatment-naïve newly diagnosed SCLC. CD56+ CTC were isolated from whole blood samples, at diagnosis and relapse after first-line treatment and submitted to whole-exome-sequencing (WES). Phenotypic study confirms tumor lineage and tumorigenic properties of isolated cells for the 4 patients analyzed with WES. WES of CD56+ CTC and matched tumor biopsy reveal genomic alteration frequently impaired in SCLC. At diagnosis CD56+ CTC were characterized by a high mutation load, a distinct mutational profile and a unique genomic signature, compared to match tumors biopsies. In addition to classical pathways altered in SCLC, we found new biological processes specifically affected in CD56+ CTC at diagnosis. High numeration of CD56+ CTC (> 7/ml) at diagnosis was associated with ES-SCLC. Comparing CD56+ CTC isolated at diagnosis and relapse, we identify differentially altered oncogenic pathways (e.g. DLL3 or MAPK pathway). We report a versatile method of CD56+ CTC detection in SCLC. Numeration of CD56+ CTC at diagnosis is correlated with disease extension. Isolated CD56+ CTC are tumorigenic and show a distinct mutational profile. We report a minimal gene set as a unique signature of CD56+ CTC and identify new affected biological pathways enriched in EpCAM-independent isolated CTC in SCLC.

History of arthropod-borne virus infections in French Polynesia.

In French Polynesia, arthropod-borne diseases are major public health problems. From the mid-1940s, the four serotypes of dengue virus (DENV-1 to -4) have caused 15 epidemics of variable severity. In 2013, for the first time, a sustained co-circulation of two different DENV serotypes (DENV-1 and -3) was reported. The same year, Zika virus (ZIKV) caused the largest outbreak ever recorded at that time. Severe neurologic complications in adults, including Guillain-Barré syndrome and central nervous system malformations in newborns and foeteuses, such as microcephaly, were reported, and a causal link with ZIKV infection was established. In addition to mosquito-borne transmission, the potential for perinatal, sexual and blood-transfusion transmission of ZIKV was demonstrated. In 2014, chikungunya virus (CHIKV) caused an explosive outbreak. Series of Guillain-Barré syndrome temporally associated with the CHIKV epidemic were reported. Except for DENV, ZIKV and CHIKV, no other arboviruses have been detected so far, but serologic evidence suggested the past silent circulation of Ross River virus. From May 2015 DENV-1 has been the only arbovirus transmitted in French Polynesia, but the reemergence of DENV-2 is highly expected since the detection of two autochthonous cases of DENV-2 infection in June 2018.

Amustaline (S-303) treatment inactivates high levels of Chikungunya virus in red-blood-cell components.

BACKGROUND AND OBJECTIVES: Chikungunya virus (CHIKV) infections have been reported in all continents, and the potential risk for CHIKV transfusion-transmitted infections (TTIs) was demonstrated by the detection of CHIKV RNA-positive donations in several countries. TTIs can be reduced by pathogen inactivation (PI) of blood products. In this study, we evaluated the efficacy of amustaline and glutathione (S-303/GSH) to inactivate CHIKV in red-blood-cell concentrates (RBCs). MATERIAL AND METHODS: Red-blood-cells were spiked with high level of CHIKV. Infectious titres and RNA loads were measured before and after PI treatment. Residual CHIKV infectivity was also assessed after five successive cell culture passages. RESULTS: The mean CHIKV titres in RBCs before inactivation was 5·81 ± 0·18 log10 50% tissue culture infectious dose (TCID50 )/mL, and the mean viral RNA load was 10·49 ± 0·15 log10 genome equivalent (GEq)/mL. No CHIKV TCID was detected after S-303 treatment nor was replicative CHIKV particles and viral RNA present after five cell culture passages of samples obtained immediately after S-303 treatment. CONCLUSION: Chikungunya virus was previously shown to be inactivated by the PI technology using amotosalen and ultraviolet A light for the treatment of plasma and platelets. This new study demonstrates that S-303/GSH can inactivate high titres of CHIKV in RBCs.

Impact of the French 3rd and 4th generation pill scare in women seeking termination of pregnancy.

INTRODUCTION: The aim of this study was to evaluate changes in the contraceptive profile of women seeking termination of pregnancy following the debate on 3rd and 4th generation pills in France in 2012. MATERIALS AND METHODS: Single-center case-control study comparing the attitude to contraception before (between 02/15/2012 and 07/16/2012) and after the debate (between 02/25/2013 and 06/24/2013). RESULTS: A total of 291 patients consulted before and 601 after the debate. We showed that there were more students (+9.5%), more single women (+8.3%) and fewer working women (-7.7%) in the cohort after the debate. After the termination procedure, prescriptions for long-acting reversible contraceptive (LARC) methods increased (+7.8%, P=0.03), in particular in patients aged 25 or younger, including nulliparous (+12.6%, P=0.02). CONCLUSION: The media alert about the pill led to a change in the contraceptive standard in the post-abortion period and altered patient profiles. An increase was notably observed in certain vulnerable populations (high school students, unemployed and single women). It remains to be seen whether these changes are transient or permanent.

Temporal and spatial discordance of programmed cell death-ligand 1 expression and lymphocyte tumor infiltration between paired primary lesions and brain metastases in lung cancer.

BACKGROUND: The dynamics of PD-L1 expression may limit its use as a tissue-based predictive biomarker. We sought to expand our understanding of the dynamics of PD-L1 expression and tumor-infiltrating lymphocytes (TILs) in patients with lung cancer-related brain metastases. EXPERIMENTAL DESIGN: Paired primary lung cancers and brain metastases were identified and assessed for PD-L1 and CD3 expression by immunohistochemistry. Lesions with 5% or greater PD-L1 expression were considered positive. Agreement statistics and the χ(2) or Fisher's exact test were used for analysis. RESULTS: We analyzed 146 paired lesions from 73 cases. There was disagreement of tumor cell PD-L1 expression in 10 cases (14%, κ = 0.71), and disagreement of TIL PD-L1 expression in 19 cases (26%, κ = 0.38). Most paired lesions with discordant tumor cell expression of PD-L1 were obtained 6 or more months apart. When specimens were categorized using a proposed tumor microenvironment categorization scheme based on PD-L1 expression and TILs, there were significant changes in the classifications because many of the brain metastases lacked either PD-L1 expression, tumor lymphocyte infiltration or both even when they were present in the primary lung cancer specimens (P = 0.009). CONCLUSIONS: We identified that there are significant differences between the tumor microenvironment of paired primary lung cancers and brain metastases. When physicians decide to treat patients with lung cancer with a PD-1 or PD-L1 inhibitor, they must do so in the context of the spatial and temporal heterogeneity of the tumor microenvironment.

ASCL1 and RET expression defines a clinically relevant subgroup of lung adenocarcinoma characterized by neuroendocrine differentiation.

ASCL1 is an important regulatory transcription factor in pulmonary neuroendocrine (NE) cell development, but its value as a biomarker of NE differentiation in lung adenocarcinoma (AD) and as a potential prognostic biomarker remains unclear. We examined ASCL1 expression in lung cancer samples of varied histologic subtype, clinical outcome and smoking status and compared with expression of traditional NE markers. ASCL1 mRNA expression was found almost exclusively in smokers with AD, in contrast to non-smokers and other lung cancer subtypes. ASCL1 protein expression by immunohistochemical (IHC) analysis correlated best with synaptophysin compared with chromogranin and CD56/NCAM. Analysis of a compendium of 367 microarray-based gene expression profiles in stage I lung adenocarcinomas identified significantly higher expression levels of the RET oncogene in ASCL1-positive tumors (ASCL1(+)) compared with ASCL1(-) tumors (q-value <10(-9)). High levels of RET expression in ASCL1(+) but not in ASCL1(-) tumors was associated with significantly shorter overall survival (OS) in stage 1 (P=0.007) and in all AD (P=0.037). RET protein expression by IHC had an association with OS in the context of ASCL1 expression. In silico gene set analysis and in vitro experiments by ASCL1 shRNA in AD cells with high endogenous expression of ASCL1 and RET implicated ASCL1 as a potential upstream regulator of the RET oncogene. Also, silencing ASCL1 in AD cells markedly reduced cell growth and motility. These results suggest that ASCL1 and RET expression defines a clinically relevant subgroup of ∼10% of AD characterized by NE differentiation.

Three-dimensional helical computed tomography in prenatal diagnosis of fetal skeletal dysplasia.

OBJECTIVES: (1) To study the use and diagnostic value, as a complement to ultrasound, of helical computed tomography (helical CT) to differentiate normal fetuses from cases of skeletal dysplasia; (2) to define the most relevant indications for helical CT; and (3) to evaluate its diagnostic performance with respect to radiological criteria considered discriminatory. METHODS: This was a retrospective study from 2005 to 2008 in 67 pregnant women who underwent helical CT after 26 weeks of gestation for suspected fetal skeletal dysplasia due to fetal shortened long bones on ultrasound (≤ 10(th) percentile), either alone or associated with other bone abnormalities. The results were compared with pediatric examinations in 41 cases and with fetal autopsy findings after elective termination of pregnancy in the others. RESULTS: Helical CT had a sensitivity of 82%, specificity of 91% and positive and negative predictive values of 90% and 83%, respectively, for diagnosis of fetal skeletal dysplasia. An etiological diagnosis that had not been suspected at ultrasound was specified in 15% of cases and diagnoses suspected at ultrasound were confirmed in 24% and discounted in 43% of cases. The prevalence of skeletal dysplasia was increased in cases of micromelia < 3(rd) percentile or if there was a combination of bone signs. Helical CT showed 69% sensitivity in identifying individual predefined pathological bone signs which were confirmed on fetal autopsy findings. CONCLUSION: Helical CT is a key examination, in combination with ultrasound, in the diagnosis of fetal skeletal dysplasia from 26 weeks of gestation. It should be reserved for cases with severe micromelia below the 3(rd) percentile and for those with micromelia ≤ 10(th) percentile associated with another bone sign. A checklist of discriminatory signs is proposed.

Integrin α9ß1 promotes malignant tumor growth and metastasis by potentiating epithelial-mesenchymal transition.

The integrin α9ß1 binds a number of extracellular matrix components to mediate cell adhesion, migration and tissue invasion. Although expressed in a variety of normal human cells including endothelium, it is also expressed in cancer cells. We have previously shown that α9ß1 binds VEGF-A to facilitate angiogenesis, an important component of the tumor microenvironment. As α9ß1 induces accelerated cancer cell migration, we wished to determine what role it played in cancer growth and metastasis. In this study, we show that α9ß1 expression induces molecular changes consistent with epithelial-mesenchymal transition. In addition, we found that α9ß1 forms a tri-partite protein complex with ß-catenin and E-cadherin, which dissociates following integrin activation and subsequent src and ß-catenin phosphorylation. These findings were consistent in cells in which: α9ß1 was exogenously over-expressed, or when its expression was suppressed in cancer cells endogenously expressing α9ß1. These in vitro results are biologically significant as α9ß1-expressing cancer cells induce greater tumor growth and metastases in mice as compared to the cells without α9ß1 expression or when integrin expression is suppressed. Furthermore, integrin α9ß1 is expressed in primary human small cell lung cancer and patients having a high expression of α9ß1 demonstrated significantly worse long-term survival compared with patients with low α9ß1 expression. These findings highlight a novel mechanism of integrin α9ß1 function in human cancer.

Iron excess limits HHIPL-2 gene expression and decreases osteoblastic activity in human MG-63 cells.

UNLABELLED: In order to understand mechanisms involved in osteoporosis observed during iron overload diseases, we analyzed the impact of iron on a human osteoblast-like cell line. Iron exposure decreases osteoblast phenotype. HHIPL-2 is an iron-modulated gene which could contribute to these alterations. Our results suggest osteoblast impairment in iron-related osteoporosis. INTRODUCTION: Iron overload may cause osteoporosis. An iron-related decrease in osteoblast activity has been suggested. METHODS: We investigated the effect of iron exposure on human osteoblast cells (MG-63) by analyzing the impact of ferric ammonium citrate (FAC) and iron citrate (FeCi) on the expression of genes involved in iron metabolism or associated with osteoblast phenotype. A transcriptomic analysis was performed to identify iron-modulated genes. RESULTS: FAC and FeCi exposure modulated cellular iron status with a decrease in TFRC mRNA level and an increase in intracellular ferritin level. FAC increased ROS level and caspase 3 activity. Ferroportin, HFE and TFR2 mRNAs were expressed in MG-63 cells under basal conditions. The level of ferroportin mRNA was increased by iron, whereas HFE mRNA level was decreased. The level of mRNA alpha 1 collagen type I chain, osteocalcin and the transcriptional factor RUNX2 were decreased by iron. Transcriptomic analysis revealed that the mRNA level of HedgeHog Interacting Protein Like-2 (HHIPL-2) gene, encoding an inhibitor of the hedgehog signaling pathway, was decreased in the presence of FAC. Specific inhibition of HHIPL-2 expression decreased osteoblast marker mRNA levels. Purmorphamine, hedgehog pathway activator, increased the mRNA level of GLI1, a target gene for the hedgehog pathway, and decreased osteoblast marker levels. GLI1 mRNA level was increased under iron exposure. CONCLUSION: We showed that in human MG-63 cells, iron exposure impacts iron metabolism and osteoblast gene expression. HHIPL-2 gene expression modulation may contribute to these alterations. Our results support a role of osteoblast impairment in iron-related osteoporosis.

18F-fluoro-2-deoxy-D-glucose positron emission tomography and positron emission tomography/computed tomography imaging of malignant pleural mesothelioma.

Malignant pleural mesothelioma (MPM) is the most common primary pleural tumor and its incidence is rising. Its diagnosis, staging and response assessment are challenging for imaging. Integrated positron emission tomography (PET)/CT increases the accuracy of overall staging in patients with mesothelioma and improves the selection of patients for curative surgical resection. It is particularly useful in identifying occult distant metastases. It may be used to predict prognosis and to assess the metabolic response to therapy.

Return to play after sports concussion in elite and non-elite athletes?

OBJECTIVE: To examine the published literature relating to the difference in concussion management strategies between elite and non-elite athletes. DESIGN: Systematic literature review of concussion management. INTERVENTION: Pubmed, Medline, Psych Info, Cochrane Library and Sport Discus databases were reviewed using the MeSH keywords brain concussion and mild traumatic brain injury, combined with athletic injuries. Each were then refined by adding the keyword "return to play" (RTP). English language and human studies only were assessed. RESULTS: For the Medline search, using "brain concussion" as a keyword, 4319 articles were found; this was decreased to 111 when RTP was used to refine the search. When "mild traumatic brain injury" was used, 2509 articles were found; this decreased to 39 when RTP was used to refine the search. Following initial review, these articles form the basis of the discussion below. CONCLUSIONS: The non-elite athlete may not have the same resources available as the elite athlete (such as the presence of trained medical staff during practice and competition, a concussion programme as part of sideline preparedness, the benefit of neuropsychological or postural testing, as well as consultants with expertise in concussion readily available) and as a result will generally be managed more conservatively. Younger athletes often have a greater incidence of concussion with longer recovery time frames; however, they are often managed with less expertise and with limited resources.

Injury surveillance in multi-sport events: the International Olympic Committee approach.

BACKGROUND: The protection of athletes' health by preventing injuries is an important task for international sports federations. Standardised injury surveillance provides not only important epidemiological information, but also directions for injury prevention, and the opportunity for monitoring long-term changes in the frequency and circumstances of injury. Numerous studies have evaluated sports injuries during the season, but few have focused on injuries during major sport events such as World Championships, World Cups or the Olympic Games. OBJECTIVES: To provide an injury surveillance system for multi-sports tournaments, using the 2008 Olympic Games in Beijing as an example. METHODS: A group of experienced researchers reviewed existing injury report systems and developed a scientific sound and concise injury surveillance system for large multi-sport events. RESULTS: The injury report system for multi-sport events is based on an established system for team sports tournaments and has proved feasible for individual sports during the International Association of Athletics Federations World Championships in Athletics 2007. The most important principles and advantages of the system are comprehensive definition of injury, injury report by the physician responsible for the athlete, a single-page report of all injuries, and daily report irrespective of whether or not an injury occurred. Implementation of the injury surveillance system, all definitions, the report form, and the analysis of data are described in detail to enable other researchers to implement the injury surveillance system in any sports tournament. CONCLUSION: The injury surveillance system has been accepted by experienced team physicians and shown to be feasible for single-sport and multi-sport events. It can be modified depending on the specific objectives of a certain sport or research question; however, a standardised use of injury definition, report forms and methodology will ensure the comparability of results.

First-trimester ultrasound diagnosis of skeletal dysplasia associated with increased nuchal translucency thickness.

A series of five cases of skeletal dysplasia is reported in which the diagnosis was reached at the 11-14-week routine ultrasound examination in our referral center. All five cases had increased nuchal translucency thickness (NT) associated with bone abnormalities. We review the current literature on skeletal dysplasia in the first trimester of pregnancy associated with increased NT.

Nurses' views on ease of patient care in postoperative pain management.

The fentanyl HCl iontophoretic transdermal system (ITS) is a compact, needle-free, pre-programmed patient-controlled analgesic system that was developed to address limitations to existing therapies for postoperative pain management. A randomized, controlled trial was conducted in 11 European countries to evaluate the efficacy and safety of postoperative pain control using fentanyl ITS compared with a standard regimen of morphine provided by an intravenous patient-controlled analgesia (IV PCA) pump. This article summarizes results from Nurse Ease-of-Care Questionnaires which were completed to assess the convenience and ease of use of each pain management modality from the perspective of the nurse. Nurses' ratings of patient-care tasks associated with each pain management system were significantly more favourable for fentanyl ITS than for morphine IV PCA. These findings suggest that nurses consider fentanyl ITS to be easier to use than morphine IV PCA.

Prenatal diagnosis and normal outcome of a 46,XX/46,XY chimera: a case report.

The phenotypic spectrum of 46,XX/46,XY chimeric patients is variable. It ranges from normal male or female genitalia to different degrees of ambiguous genitalia. Chimerism results from the amalgamation of two different zygotes in a single embryo, whereas mosaicism results from a mitotic error in a single zygote. Several other mechanisms resulting in a chimera have been discussed in the literature. Here, we report on a new case of chimerism (46,XX/46,XY) diagnosed at 17 weeks' gestation on amniocentesis performed because of advanced maternal age. Ultrasound examination revealed normal female external genitalia, and a healthy baby girl was delivered at term. We used polymorphic markers spanning the X chromosome and several autosomes in order to identify the genetic mechanism involved. Mosaicism was excluded because of the presence of 3 alleles at 11 autosomal and 4 X chromosome loci. On autosomes, the origin of this third allele was maternal for two pericentromeric markers (located on 2p11.2 band and 8p11.2 band), paternal for six markers and paternal or maternal for the other three markers. On the X chromosome, the origin of the third allele was maternal for all four markers. Thus, two different paternal and maternal haploid sets were observed. These results are compatible with a tetragametic chimera.