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.

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.

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.

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.

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.

Tubulin is phosphorylated at tyrosine by pp60c-src in nerve growth cone membranes.

We show here that tubulin is the major in vivo substrate of the tyrosine-specific protein kinase pp60c-src in nerve growth cone membranes. Phosphotyrosine antibodies were used to demonstrate phosphotyrosyl residues in a subpopulation of alpha- and beta-tubulin that was highly enriched in a subcellular fraction of growth cone membranes from fetal rat brain. The presence of phosphotyrosine-modified isoforms of alpha- and beta-tubulin in vivo was confirmed by 32p labeling of rat cortical neurons in culture. Tubulin in growth cone membranes was phosphorylated at tyrosine in endogenous membrane phosphorylation reactions (0.068 mol phosphotyrosine/mol alpha-tubulin and 0.045 mol phosphotyrosine/mol beta-tubulin), and phosphorylation was specifically inhibited by antibodies directed against pp60c-src, which is localized in the growth cone membranes. pp60c-src was capable of directly phosphorylating tubulin as shown in immune complex kinase assays with purified brain tubulin. Phosphopeptide mapping revealed a limited number of sites of tyrosine phosphorylation in alpha- and beta-tubulin, with similar phosphopeptides observed in vivo and in vitro. These results reveal a novel posttranslational modification of tubulin that could regulate microtubule dynamics at the growth cone.

LSD: no teratogenic action in rats, mice, and hamsters.

Lysergic acid diethylamide tartrate was given to 98 pregnant rats, 67 mice, and 22 hamsters as a single dose of 5 to 500 micrograms per kilogram of body weight per day either at the beginning of gestation or during the period of organogenesis. Examination of the 1003 rat fetuses, 521 mouse fetuses, and 189 hamster fetuses obtained failed to prove any abortifacient, teratogenic, or growth-depressing effects.

The Eocene/Oligocene boundary event in the deep sea.

Analysis of middle Eocene to early, Oligocene calcareous and siliceous microfossils shows gradual biotic changes with no massive extinction event across the Eocene/Oligocene boundary. Biotic changes in the late Paleogene appear to reflect changing paleoclimatic and paleoceanographic conditions and do not support suggestions of a catastrophic biotic event caused by a bolide impact at the Eocenel Oligocene boundary.

Laminated diatomaceous sediments from the guaymas basin slope (central gulf of california): 250,000-year climate record.

During Deep Sea Drilling Project-International Program of Ocean Drilling leg 64, December 1978 to January 1979, the initial test of the Deep Sea Drilling Project's hydraulic piston corer obtained an almost undisturbed section from a 152-meter hole into the sediments of the oxygen minimum zone at a depth of 655 meters along the Guaymas slope in the central Gulf of California. The section records variations in climate, productivity, and circulation for more than 250,000 years of Late Pleistocene to Holocene history with recordings of seasonal variations in these parameters in the laminated sections.

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.

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.

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.

Phenotypic spectrum of CHARGE syndrome in fetuses with CHD7 truncating mutations correlates with expression during human development.

BACKGROUND: The acronym CHARGE refers to a non-random cluster of malformations including coloboma, heart malformation, choanal atresia, retardation of growth and/or development, genital anomalies, and ear anomalies. This set of multiple congenital anomalies is frequent, despite rare patients with normal intelligence, and prognosis remains poor. Recently, CHD7 gene mutations have been identified in CHARGE patients; however, the function of CHD7 during development remains unknown. METHODS: We studied a series of 10 antenatal cases in whom the diagnosis of CHARGE syndrome was suspected, considering that a careful pathological description would shed light on the CHD7 function during development. CHD7 sequence analysis and in situ hybridisation were employed. RESULTS: The diagnosis of CHARGE syndrome was confirmed in all 10 fetuses by the identification of a CHD7 heterozygous truncating mutation. Interestingly, arhinencephaly and semi-circular canal agenesis were two constant features which are not included in formal diagnostic criteria so far. In situ hybridisation analysis of the CHD7 gene during early human development emphasised the role of CHD7 in the development of the central nervous system, internal ear, and neural crest of pharyngeal arches, and more generally showed a good correlation between specific CHD7 expression pattern and the developmental anomalies observed in CHARGE syndrome. CONCLUSIONS: These results allowed us to further refine the phenotypic spectrum of developmental anomalies resulting from CHD7 dysfunction.

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.

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.

Overview of concussion consensus statements since 2000.

More refereed publications on sports-related concussion have appeared since 2000 than in all previous years combined. Three international consensus statements, documents from the National Athletic Trainers' Association (NATA) and the American College of Sports Medicine (ACSM), and entire issues of the Clinical Journal of Sport Medicine and the Journal of Athletic Training have been devoted to this subject. The object of this article is to critique the consensus statements and NATA and ACSM documents, pointing out areas of controversy.

A kinetic study of the inhibition of human and bovine trypsins and chymotrypsins by the inter-alpha-inhibitor from human plasma.

Human plasma inter-alpha-inhibitor forms 1:1 inactive complexes with human and bovine trypsins (EC 3.4.21.4) and chymotrypsins (EC 3.4.21.1). The association and dissociation rate constants as well as the equilibrium dissociation constants (Ki) of the complexes formed of inter-alpha-inhibitor and the four proteases have been measured. The most stable complexes are those formed with the bovine enzymes. For instance, Ki = 2.1-10-11 M for bovine trypsin whereas Ki = 1.2 - 10-8 M for human trypsin. Whatever the species, the complexes formed with the chymotrypsins are less stable than those formed with the trypsins.

Monoclonal antibodies as probes for the transmembrane structure of neutral endopeptidase 24.11 ('enkephalinase').

The neutral endopeptidase (EC 3.4.24.11) ('enkephalinase') is a membrane-bound metalloendopeptidase that is present in large amounts in the microvilli of the kidney proximal tubules. By immunizing mice with purified rabbit kidney brush-border membranes, we have obtained four different monoclonal antibodies that recognize this enzyme in dot-blot and Western-blot assays and can be used for immunoprecipitation of neutral endopeptidase from crude kidney solubilizates. One of these monoclonal antibodies (2B12) allows the labeling of proximal tubule cells with colloidal gold particles. This monoclonal antibody also binds to native brush-border membrane vesicles (which are mostly in the right-side-out configuration) and recognizes an epitope which is destroyed after reduction and alkylation of the protein. By contrast, all three other monoclonal antibodies (21G10, 23B11 and 22E2) compete for another epitope of neutral endopeptidase that is not exposed at the extracytoplasmic surface either in intact cells or in sealed brush-border vesicles. Permeabilization of the vesicles with digitonin, however, restores the full binding activity. Binding of these antibodies is not altered by prior reduction and alkylation of the protein. Taken together, these results strongly suggest that the 2B12 monoclonal antibody binds a conformational epitope located on the ectodomain of the enzyme, whereas the three others (21G10, 23B11 and 22E2) bind to a common or to overlapping epitopes located on the cytosolic domain. These results also demonstrate unambiguously the transmembrane nature of neutral endopeptidase.

Duration of smoking abstinence as a predictor for non-small-cell lung cancer survival in women.

BACKGROUND: Previous studies have attempted to investigate the impact of smoking cessation on lung cancer survival but have been limited by small numbers of former smokers and incomplete data. METHODS: Over a six-year period, 5229 patients with non-small-cell lung cancer (NSCLC) and small-cell lung cancer (SCLC) were enrolled in a prospective cohort of whom 2052 were former smokers. Patient's characteristics were obtained from medical records and a baseline interview. Vital status was determined through multiple sources. Cox proportional hazards models were used to estimate the effect of smoking abstinence on post-diagnosis mortality. RESULTS: For all patients with NSCLC, the median survival among never, former, and current smokers was 1.4 years, 1.3 years, and 1.1 years, respectively (P < 0.01). Female NSCLC patients had a significantly lower risk of mortality with a longer duration of smoking abstinence (RR per 10 years of smoking abstinence = 0.85; 95% CI: 0.75, 0.97). No effect of smoking abstinence on mortality was observed for women with SCLC or for men with either histologic group. CONCLUSIONS: The identification of smoking history as a prognostic factor in lung cancer survival supports previous research suggesting a direct biologic effect of smoking on survival. However, this effect may vary by sex and type of lung cancer.

Thrombomodulin promoter mutations, venous thrombosis, and varicose veins.

We analyzed the distal promoter region of the thrombomodulin (TM) gene (nucleotides -300 to -2052) in subjects from the Paris Thrombosis Study (PATHROS), a French case-control study of venous thrombosis, to identify polymorphisms that might modify TM gene expression. Eight novel mutations were found in the 40 DNA samples initially screened. Two of these mutations (-1748G/C and -1208/-1209 del TT) were frequent. One rare transition (-1166G/A) might have functional consequences owing to its position. These 3 mutations were screened for in the entire study population of 327 patients and 398 controls. None of the 3 was significantly associated with thrombosis. Interestingly, the -1208/-1209 TT deletion was associated with varicose veins in the patients. This mutation was in tight linkage disequilibrium with the +1418 C/T change in the coding sequence, a known polymorphism that predicts an Ala 455 Val substitution in the sixth epidermal growth factor-like TM module, a domain previously implicated in the proliferative functions of TM. This linkage suggests that the Ala 455 Val mutation may promote changes in these functions and thus be involved in varicose vein formation.