Emotional hyper-reactivity and cardiometabolic risk in remitted bipolar patients: a machine learning approach.

OBJECTIVE: Remitted bipolar disorder (BD) patients frequently present with chronic mood instability and emotional hyper-reactivity, associated with poor psychosocial functioning and low-grade inflammation. We investigated emotional hyper-reactivity as a dimension for characterization of remitted BD patients, and clinical and biological factors for identifying those with and without emotional hyper-reactivity. METHOD: A total of 635 adult remitted BD patients, evaluated in the French Network of Bipolar Expert Centers from 2010-2015, were assessed for emotional reactivity using the Multidimensional Assessment of Thymic States. Machine learning algorithms were used on clinical and biological variables to enhance characterization of patients. RESULTS: After adjustment, patients with emotional hyper-reactivity (n = 306) had significantly higher levels of systolic and diastolic blood pressure (P < 1.0 × 10-8 ), high-sensitivity C-reactive protein (P < 1.0 × 10-8 ), fasting glucose (P < 2.23 × 10-6 ), glycated hemoglobin (P = 0.0008) and suicide attempts (P = 1.4 × 10-8 ). Using models of combined clinical and biological factors for distinguishing BD patients with and without emotional hyper-reactivity, the strongest predictors were: systolic and diastolic blood pressure, fasting glucose, C-reactive protein and number of suicide attempts. This predictive model identified patients with emotional hyper-reactivity with 84.9% accuracy. CONCLUSION: The assessment of emotional hyper-reactivity in remitted BD patients is clinically relevant, particularly for identifying those at higher risk of cardiometabolic dysfunction, chronic inflammation, and suicide.

Differentiation of human dendritic cell subsets for immune tolerance induction.

OBJECTIVES: Since no further progress was achieved, in order to improve the long-term organ transplantation outcome, the immune tolerance appears as an interesting therapeutic goal. Dendritic cells (DCs) are specialized cells participating in the homeostasis of the immune response. Moreover, subsets of DCs, identified in humans, appear to have their respective competences in immune response modulation. Our objective is to purify from PBMC or to differentiate DC subsets from monocytes using several strategies and evaluate their IL10 secretion. METHODS: CD14+ cells were purified from peripheral blood mononuclear cell (PBMC) by affinity beads and cultured with cytokines up to 7 days. The pDCs were purified with anti-BDCA-2 beads from PBMC fraction enriched by Percoll® gradient. The moDCs, pDCs and moLCs subsets were analyzed by phenotype labelling and FACS analyses and IL10 secretion measured by ELISA. RESULTS: The moDCs were characterized by the CD209 expression and a lower expression of CD1a markers. Expression of CD207 and CD1a markers characterized moLCs and CD123+/BDCA-2+ pDCs. Variable IL-10 secretions were shown between the three DC subsets, both at basal and activated levels. CONCLUSIONS: As the several DC populations studied have different capacities of IL-10 synthesis, they might play, among others, distinct roles in the induction of immune tolerance.

BRAF mutation in papillary thyroid carcinoma: predictive value for long-term prognosis and radioiodine sensitivity.

OBJECTIVES: BRAF pV600E mutation is the most common oncogenic event and the most specific mutation for papillary thyroid carcinoma (PTC). Many studies over the last decade have shown a direct relationship between BRAF mutation and aggressive tumour characteristics, resulting in poor prognosis. However, several recent studies have suggested that BRAF mutation is not associated with poor prognosis of PTC. The present study was designed to evaluate the association between BRAF mutation with clinicopathological factors and tumour recurrence. MATERIAL AND METHODS: In this retrospective study, BRAF mutation status was examined by direct sequencing on paraffin-embedded tumour specimens from 46 patients undergoing surgery for PTC in our institution from 1985 to 2000. The relationship between BRAF mutation and gender, advanced age, extrathyroid extension, multifocal tumour, cervical lymph node metastasis, tumour size and advanced pT stage of PTC and its predictive role for the risk of tumour recurrence were investigated with a median follow-up of 10.1 (±6.5)years. RESULTS: BRAF mutation was detected in 20 of the 46 patients (43.5%) included in the study. No statistically significant correlation was demonstrated between the presence of BRAF mutation and the various clinicopathological factors studied. No significant difference in tumour recurrence rate or radioiodine sensitivity was observed between the two subgroups: mutant BRAF and wild-type BRAF. CONCLUSION: Although BRAF mutation appears to play a role in local tumour progression, it is not a risk factor for poor prognosis or tumour recurrence in PTC.

[Amniotic fluid embolism during curettage for a pregnancy arrest. Case report]. / Embolie amniotique lors d'un curetage pour une grossesse arrêtée. A propos d'un cas.

Amniotic fluid embolism is always a serious complication during the peripartum period. We report the case of an amniotic fluid embolism during curettage for a pregnancy arrest at 13 weeks. The diagnosis was confirmed by the presence of epithelial cells into the maternal blood.

Impact of a polymorphism in the IL-12p40 gene on the outcome of kidney transplantation.

A number of factors interfere with the outcome of renal transplantation. Revealing genetic factors that impact on graft outcome may have consequences for clinical practice. Interleukin-12 (IL-12), by stimulating interferon gamma (IFNgamma) production, plays a crucial role in immune responses against both graft and viral agents. An A-to-C single nucleotide polymorphism (SNP) within the 3'-untranslated region (3'UTR) of the IL-12p40 gene has been reported to be both functionally and clinically relevant. Since the impact of this SNP on kidney graft outcome has never been reported, we investigated the impact of the 3'UTR polymorphism on clinical events after transplantation among 253 kidney recipients transplanted between 1995 and 2003. The polymorphism was genotyped using the restriction fragment length polymorphism method. Our results showed that the 3'UTR polymorphism affected neither graft survival (P = .768) nor the occurrence of delayed graft function (DGF; P = .498). C allele carriers in our study displayed more acute rejections in the first year than patients with the A/A genotype, but it did not reach statistical significance (P = .108). In contrast, the C allele appeared to be a significant risk factor for cytomegalovirus infection (odds ratio = 1.77; P = .027). In conclusion, IL12B 3'UTR polymorphism did not affect graft survival, DGF, or acute rejection episodes, but had an impact on the occurrence of cytomegalovirus infection.

Anti-CD25 antibodies decrease the ability of human dendritic cells to prime allogeneic CD4 T cells.

Anti-CD25 monoclonal antibodies are largely used in clinical transplantation to prevent acute allograft rejection episodes. Although their effects on T lymphocytes have been extensively studied, their impact on human dendritic cells (DCs) has been less reported. Furthermore, the role of the interleukin-2 in DC functions has not yet been fully elucidated. In this study, we observed that stimulation of human monocyte-derived DCs with lipopolysa ccharide or CD40L strongly induced the expression of CD25. We showed that pretreatment of DC with anti-CD25 diminished their ability to prime T-helper cells. In contrast, humanized anti-CD25 monoclonal antibodies did not affect the up-regulation of CD86, CD80, CD83, HLA-DR, or CD40 induced by lipopolysaccharide stimulation. This study supported previously unrecognized effects of anti-CD25 monoclonal antibodies on DCs that may contribute to their clinical efficacy.

Mycophenolic Acid Inhibits p38 Mitogen-Activated Protein Kinase in Human Monocyte-Derived Dendritic Cells Stimulated by Lipopolysaccharide.

Dendritic cell (DC) maturation, a crucial stage in the immune response, can be induced by various stimuli, such as lipopolysaccharide (LPS). Maturation signals trigger up-regulation of costimulatory molecule expression, increasing the ability of DCs to prime T helper cells. We and others have previously reported that mycophenolic acid (MPA) inhibits DC maturation and activation. However, the mechanisms remain unknown. The primary effect of MPA is inhibition of inosine monophosphate dehydrogenase (IMPDH), an enzyme involved in the de novo synthesis of guanosine nucleotide. The process of DC maturation is highly dependent on mitogen-activated protein kinase (MAPK) phosphorylation, especially p38MAPK. We therefore decided to study whether MPA affects these processes. Human monocyte-derived DCs were activated by LPS in the presence or absence of MPA. To assess whether the depletion of guanine affected p38MAPK phosphorylation, increasing doses of exogenous guanosine were added before stimulation. The results by flow cytometry showed that MPA inhibited p38MAPK phosphorylation by 25%. Interestingly, exogenous guanosine did not reverse the MPA inhibition. Our results suggested that MPA inhibits p38MAPK activity independent of IMPDH in human DCs. This effect of MPA may explain its capacity to inhibit maturation marker expression on DCs.

Induction of porcine regulatory cells by mycophenolic Acid-treated dendritic cells.

Tolerance induction in murine allogeneic transplantation is relatively easy, often by induction of regulatory T cells (Treg). Unfortunately, the implementation of these models in clinical situations has not yielded reliable protocols of tolerance induction in humans. Our project sought to create a preclinical model of tolerance induction in large animals. Our current efforts seek to induce and characterize porcine Treg, obtaining dendritic cells (DC) able to preferentially stimulate them. DCs were differentiated from blood monocytes with porcine recombinant interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) for 6 days. These DCs were then stimulated by human CD40 ligand-transfected L cells with or without mycophenolic acid (MPA) for 48 hours. We analyzed surface marker expression, cytokine synthesis, and ability to stimulate allogeneic peripheral blood mononuclear cells (PBMC). The porcine lymphocytes underwent 4 rounds of 1-week stimulation with allogeneic DC treated or not with MPA. At the end of this coculture we analyzed their capacity to suppress allogeneic PBMC proliferation induced by mature DC. Our results showed that porcine DCs pretreated with MPA display a low expression of B7 costimulatory molecules, produce low levels of IL-12, and induce weak proliferation of allogeneic lymphocytes. Moreover, after 4 rounds of stimulation with MPA-treated DCs, PBMCs were able to inhibit an alloreactive response. These preliminary results suggested induction of a regulatory T-cell population that we are currently seeking to characterize.

Anti-CD25 antibodies affect cytokine synthesis pattern of human dendritic cells and decrease their ability to prime allogeneic CD4+ T cells.

Anti-CD25 monoclonal antibodies are widely used in clinical transplantation to prevent acute allograft rejection. Although their effects on T lymphocytes have been extensively studied, their impact on human dendritic cells (DC) has never been reported. Furthermore, the role of the IL-2 in DC functions has not yet been fully elucidated. In this study, we confirm that the stimulation of human monocyte-derived DC with LPS strongly induced the expression of CD25 and that LPS-matured DC also expressed the beta and gamma chain of the IL-2R. We also showed that adding anti-CD25 monoclonal antibodies to LPS induced a decrease in IL-12, IL-1, TNF-alpha, IL-6, and IFN-gamma production and an increase in IL-10 synthesis by DC compared with stimulation with LPS alone. Furthermore, we showed that these modifications diminished the T helper priming ability of DC and polarized the alloimmune response toward TH2. In contrast, humanized anti-CD25 monoclonal antibodies did not affect the up-regulation of CD86, CD80, CD83, HLADR, or CD40 induced upon LPS stimulation. Taken together, this study discloses some previously unrecognized effects of anti-CD25 monoclonal antibodies on DC that may contribute to their clinical efficacy. In addition, this study also shed some light on the role of the IL-2 in human DC activation.

[The script concordance test: a new evaluation method of both clinical reasoning and skills in internal medicine]. / Le test de concordance de script: un nouvel outil d'évaluation du raisonnement et de la compétence clinique en médecine interne ?

OBJECTIVE: The script concordance test is designed to evaluate knowledge organization, which constitutes a crucial parameter of clinical skills. The objective of the present study was to assess the value of a new written evaluation tool to measure clinical skills in Internal Medicine. MATERIALS AND METHODS: A 95-item examination was completed by a group of medical students (N =17), a group of residents in Family practice (N =9), a group of residents in Internal Medicine (N =5), and a group of experienced physicians in Internal Medicine (N =7). The scores obtained were compared by analysis of variance. The reliability of the test was studied by calculating Cronbach's coefficient alpha. RESULTS: The mean score was 220.3 +/-41.7 for medical students, 230.5 +/-31.7 for residents in Family practice, 274.2 +/-32.2 for residents in Internal Medicine, and 352.1 +/-22.9 for experienced physicians in Internal Medicine. The differences observed between the scores for the various groups were significant (P <0.0001). Moreover, the value of Cronbach's coefficient alpha was 0.81 in the whole examination. CONCLUSION: Our data indicate that the script concordance test may easily allow to differentiate various levels of clinical skills in Internal Medicine. Moreover, because of Cronbach's coefficient alpha as high as 0.81, our findings suggest the validity of this test in Internal Medicine.

Conducting polymers as driving electrodes for Polymer-Dispersed Liquid-Crystals display devices: on the electro-optical efficiency.

Intrinsically conducting polymer (ICP) thin films are used as driving electrodes for Polymer-Dispersed Liquid-Crystals (PDLC) display devices. In order to investigate the electro-optical efficiency of these organic electrodes, three different kinds of conducting polymers, i.e. polyaniline doped with 10-camphorsulfonic acid (PANI(HCSA)), polypyrrole doped with dodecylbenzenesulfonic acid (PPY(DBSA)), and polyethylenedioxythiophene doped with polystyrenesulfonate (PEDOT(PSS)), were prepared or purchased, and coated either on glass or plastic substrates. Optical absorption studies in the UV-Vis range of the conducting polymer-coated substrates were first performed showing the presence of conducting species for the three types of polymers. The electrical characteristics of the resulting films were measured with the four-probes technique. PANI(HCSA) exhibits a higher conductivity sigma approximately 122 S x cm(-1) (RS=1.2x10(3) Omega x (-1)) compared to PPY(DBSA) sigma approximately 2.6 S x cm(-1) (RS=150.7x10(3) Omega x (-1)), and PEDOT(PSS) sigma approximately 1.6 S x cm(-1) (RS=637.3x10(3) Omega x (-1)). It is also shown that for a given conducting polymer, its electrical conductivity decreases when a plastic substrate is used. These observations have been related to significant morphological changes observed by scanning electron microscopy (SEM). A mixture of Norland Optical Adhesive 65 and nematic liquid-crystal E7 in the weight ratio (35:65) was used as precursor of the PDLC material. Better electro-optical responses (transmission properties, drive voltages and switching times) of PDLC films were obtained for devices prepared with (PPY(DBSA))-based electrodes. The electro-optical performances of the PDLC display devices also depend on the nature of the ICP substrate used.

Prospects for a human Toxoplasma vaccine.

Human toxoplasmosis is usually benign, but may occasionally lead to severe or lethal damages when combined with immunosuppressive states or when transmitted to the fetus during pregnancy. Only a vaccine could prevent these harmful effects. The oral route is the natural portal of entry of T. gondii. A protective immune response at the mucosal level is required to kill the parasite as soon as it penetrates the intestinal barrier thus preventing toxoplasma from invading the host and settling into tissues. The probable major roles played by both CD8 T cells and antibodies, specially IgA, suggest that the best strategy would be to stimulate both the cellular and humoral arms of the mucosal immune system. Mucosal dendritic cells have been shown to induce good protection against oral toxoplasma challenge. Our hypothesis is that an acceptable and effective human vaccine would have to carry the optimized synthetic vaccine (subunit, DNA or replicon) plus an appropriate adjuvant and to target the mucosal dendritic cells by means of an inert delivery system such as polymer microparticles, which can be endocytosed by M cells of the gut or nasal-associated lymphoid tissues.

Thermophysical properties of fluorinated acrylate homopolymers: mixing and phase separation.

The thermophysical properties of fluorinated acrylate homopolymers are investigated by differential scanning calorimetry (DSC) and optical microscopy and discussed in terms of relative lengths of the fluorinated chain and the hydrocarbon spacer between the acrylate moiety and the fluorinated chain. These compounds exhibit an intrinsic microphase-separation (Isotropic+Isotropic morphology) occurring between the fluorinated chains and the acrylate polymer backbone. It is shown that the enthalpy of mixing is a function of the length of the lateral fluorocarbon chains. The thermophysical behaviour of these materials may be regarded as demixed systems exhibiting an Upper Critical Solution Temperature. The photopolymerization process of one of the monomer is studied by isothermal photocalorimetry. High acrylate double-bond conversion and fast curing rates were obtained thus demonstrating the promising use of these materials for coating and film processing applications using UV-curing techniques.

Solid-phase synthesis of a branched hexasaccharide using a highly efficient synthetic strategy.

The solid-phase synthesis of branched lacto-N-neohexaose derivative 1 occurring in human milk is described. The new building block of lactose 3 bearing the orthogonal temporary hydroxy protecting groups 9-fluorenylmethyloxycarbonyl (Fmoc) and levulinoyl (Lev) has been prepared. Its use, together with that of lactosamine donor 4, glucosamine donor 5, and O-galactosyl trichloroacetimidate 6, has enabled the preparation of hexasaccharide 22 following two different approaches in excellent overall yield (43%, 90% per step over eight steps). An additional key feature of this work is the successful use of newly prepared ester-type linker 2, having a benzylic spacer connected to the anomeric oxygen. This linker presents the advantage of producing a benzylic anomeric moiety after cleavage from the polymer support, which could be easily removed to obtain the unprotected oligosaccharide 1.

Anti-SAG1 peptide antibodies inhibit the penetration of Toxoplasma gondii tachyzoites into enterocyte cell lines.

The initial attachment of Toxoplasma tachyzoites to the target host cell is an important event in the life-cycle of the parasite and a critical stage in infection. Previous studies have shown that polyclonal antibodies directed against the major surface antigen of Toxoplasma gondii (SAG1) inhibit the infection of enterocyte cell lines. Here, we demonstrate that antibodies raised against a central peptide (V41T) of SAG1 and the SAGI protein itself are able to inhibit the infection of various cell lines by the tachyzoites. Antibodies directed against SAG1 peptides were used to define a site on the SAGI antigen that interacts with the host cell. The epitope carried by V41T was identified on the tachyzoite surface by immunofluorescence. The peptide sequence seems to be conserved in all the members of the SAGI Related Sequence family (SRS). Using undifferentiated and differentiated Caco-2 cells, we found that tachyzoites enter preferentially via the basolateral side of the cell. These findings highlight the role of the SRS family members in the mediation of host cell invasion.