Increase in subjective well-being and psychological health after application of C8-silk lipoamino acid functionalized pigments included in a foundation.

OBJECTIVE: The aim of this study was to evaluate the effects of two foundations before (D0) and after 5 days of application (D6) on psychophysiological parameters in order to compare C8-silk lipoamino acid functionalized pigments (FA) versus pure pigments (FP). METHODS: Assessment of self-esteem, stress, mood and emotion using psychological tests and evaluation of salivary cortisol concentrations were realized on 40 healthy females using a crossover study design at D0 and D6. Four saliva samples were taken on the awakening (C1), 30 min after the awakening (during the foundation application (C2)), 1 h after the foundation application (C3) and at 1900 h (C4) at DO and D6. Area under the curve was calculated in order to obtain information about the total amount of a given substance excreted in a specific time period. RESULTS: Five days of daily application of the foundation containing the pigments treated with the C8-silk lipoamino acids induced a significant increase (P < 0.001) in self-esteem and pleasant emotion (P < 0.05), and a significant decrease in general stress (P < 0.05). This was not the case after the FP application. Cortisol concentrations presented a pronounced diurnal rhythm whatever the foundation used. At DO, no significant differences were observed between the groups. At D6, cortisol concentrations measured 30 min and 1 h after the FA application were significantly lower (P < 0.05: C6.2, P < 0.05 C6.3, respectively) than those reported after FP application. AUC, a global stress response indicator, was significantly lower in FA group as compared to FP group after 5 days of application. Subjects found a decrease in tiredness signs and thought that FA has a good coverage. CONCLUSION: Our results show that incorporation of C8-silk lipoamino acid as agent of pigment functionalization brings new benefits to a foundation. Adopting a psychophysiological approach, which is not invasive to the subjects, we show the measurement of cortisol at the same time that psychological indicators provide a scientific approach to examine the beneficial effects of a cosmetic product.

OBJECTIF: Le but de cette étude randomisée croisée était d'évaluer les effets de deux fonds de teints (pigments traditionnels (FP) versus lipoaminoacides de C8 soie (FA)) avant (D0) et après 5 jours d'application (D6) sur des paramètres psychophysiologiques. METHODES: L'évaluation de l'estime de soi, du stress, de l'humeur et des émotions conjointement à l'évaluation des concentrations de cortisol salivaire ont été réalisées sur 40 femmes. Quatre prélèvements salivaires ont été effectués le matin au réveil (C1), 30 min après (pendant l'application du fond de teint (C2)), 1h après l'application du fond de teint (C3) et à 19h00 (C4) au début (D0) et en fin d'expériementation (D6). RÉSULTATS: Cinq jours d'application quotidienne du fond de teint contenant des pigments traités au lipoaminoacides de C8 soie ont induit une augmentation significative (P <0.001) de l'estime de soi et des émotions plaisantes (P <0.05), ainsi qu'une diminution significative du stress (P < 0.05). Ces résultats n'ont pas été observés après application du pigment traditionnel (FP). En début d'expérimentation (D0), aucune différence significative n'a été observée entre les groupes concernant les concentrations de cortisol salivaire, concentrations qui présentaient un rythme diurne prononcé. A D6, les concentrations de cortisol mesurées 30 minutes et 1 heure après l'application de FA étaient significativement inférieures (P < 0.05: C6.2, P < 0.05: C6.3, respectivement) à celles rapportées après l'application de FP. Les sujets ont constaté une diminution des signes de fatigue. CONCLUSION: Nos résultats montrent que l'incorporation du lipoaminoacide de C8 soie en tant qu'agent de fonctionnalisation des pigments apporte de nouveaux avantages à une base de fond de teint. L'approche psychophysiologique non invasive est une excellente approche permettant d'évaluer les effets bénéfiques d'un produit cosmétique.

Identification of T-cell epitopes from benzylpenicillin conjugated to human serum albumin and implication in penicillin allergy.

BACKGROUND: There is in vitro evidence that T cells from allergic patients react to benzylpenicillin-human serum albumin (BP-HSA) bioconjugates. Our group has recently shown the existence of naïve CD4+ T cells recognizing BP-HSA in healthy donors. However, BP-haptenated peptides from HSA participating in the immunization of allergic patients have never been identified. The purpose of the present study is to identify immunodominant BP-haptenated peptides from HSA involved in immunization of patients to BP and to refine the frequency calculation of naïve CD4+ T cells recognizing BP. METHODS: Co-cultures were established with CD4+ T cells from non-allergic donors and mature autologous dendritic cells (DCs) loaded with BP-HSA or BP-haptenated peptides from HSA. The CD4+ T-cell response specific for BP-HSA or for individual BP-haptenated peptides was measured using an interferon-γ (IFN-γ) ELISpot assay. The frequency of BP-specific CD4+ T cells was then calculated using the Poisson distribution. BP-HSA and BP-haptenated peptides recognition by allergic patients was evaluated on peripheral blood mononuclear cells (PBMCs) using a lymphocyte transformation test (LTT). RESULTS: Results showed that BP-HSA and BP-haptenated peptides were recognized by naïve T cells from 15/16 and 13/14 tested healthy donors, respectively. Most donors responded to 3 peptides with BP covalently bound on lysines 159, 212, and 525. Two of these benzylpenicilloylated peptides (lysines 159 and 525) were also found to induce PBMCs proliferation in patients with allergic reaction to penicillins. CONCLUSION: This study identifies and characterizes for the first time the BP-haptenated peptides from HSA involved in the immunization of patients to penicillins.

Low end-tidal CO2 as a real-time severity marker of intra-anaesthetic acute hypersensitivity reactions.

BACKGROUND: Prompt diagnosis of intra-anaesthetic acute hypersensitivity reactions (AHR) is challenging because of the possible absence and/or difficulty in detecting the usual clinical signs and because of the higher prevalence of alternative diagnoses. Delayed epinephrine administration during AHR, because of incorrect/delayed diagnosis, can be associated with poor prognosis. Low end-tidal CO2 (etCO2) is known to be linked to low cardiac output. Yet, its clinical utility during suspected intra-anaesthetic AHR is not well documented. METHODS: Clinical data from the 86 patients of the Neutrophil Activation in Systemic Anaphylaxis (NASA) multicentre study were analysed. Consenting patients with clinical signs consistent with intra-anaesthetic AHR to a neuromuscular blocking agent were included. Severe AHR was defined as a Grade 3-4 of the Ring and Messmer classification. Causes of AHR were explored following recommended guidelines. RESULTS: Among the 86 patients, 50% had severe AHR and 69% had a confirmed/suspected IgE-mediated event. Occurrence and minimum values of arterial hypotension, hypocapnia and hypoxaemia increased significantly with the severity of AHR. Low etCO2 was the only factor able to distinguish mild [median 3.5 (3.2;3.9) kPa] from severe AHR [median 2.4 (1.6;3.0) kPa], without overlap in inter-quartile range values, with an area under the receiver operator characteristic curve of 0.92 [95% confidence interval: 0.79-1.00]. Among the 41% of patients who received epinephrine, only half received it as first-line therapy despite international guidelines. CONCLUSIONS: An etCO2 value below 2.6 kPa (20 mm Hg) could be useful for prompt diagnosis of severe intra-anaesthetic AHR, and could facilitate early treatment with titrated doses of epinephrine. CLINICAL TRIAL REGISTRATION: NCT01637220.

OC-06 - Pro-thrombotic biomarkers in pancreatic diseases: are they specific of cancer?

INTRODUCTION: Venous thrombo-embolic events (VTE) occur frequently in patients with pancreatic cancer and contribute to elevated morbidity and mortality. Clinical risk factors for thrombosis such as cancer stage and tumor grade have been clearly identified. Recently, several biomarkers have been proposed which may help identifying cancer patients at high risk of thrombosis. Those biomarkers have been studied in heterogeneous cohorts of patients with different cancer types. AIM: To compare pro-thrombotic biomarkers in pancreatic cancer and in chronic pancreatitis to determine whether these biomarkers are related to cancer or inflammation and to validate their association with thrombotic risk in a pancreatic cancer population. MATERIALS AND METHODS: 45 patients with pancreatic cancer, 49 with intraductal papillary mucinous tumor of the pancreas (IPMN), a precancerous lesion, and 50 with chronic pancreatitis were recruited. Plasma levels of factor VIII, D-dimers, thrombin-antithrombin complexes, soluble p-selectin, tissue factor-dependent procoagulant activity of MP (TF-MP), free Tissue Factor Pathway Inhibitor (TFPI) and extracellular DNA were measured. Thrombin generation triggered by 1pM of TF was evaluated with the Calibrated Automated Thrombogram assay. RESULTS: Levels of factor VIII, D-dimers, TF-MP, TFPI and extracellular DNA were significantly higher in cancer patients compared to IPMN or chronic pancreatitis (Table 1). Patients with metastatic cancer (n=27) presented higher levels of D-dimers (mean±sd 1.77±1.28 vs 0.80±0.96 µg/ml, p=0.004) and MP-TF (54.3±53.2 vs 15.8±10.4 fM, p=0.02) compared to patients with localized lesions (n=18). Among cancer patients, 42 were followed for a median duration of 187 days (min 21-max 802 days). VTE occurred in 10 (23%) patients. All had metastatic cancer at the time of thrombosis. Only D-dimers were significantly elevated in cancer patients with VTE compared to patients without VTE (median 1.85 vs 0.7 µg/ml, p=0.02). CONCLUSIONS: Elevation of factor VIII, D-dimers, TF-MP, TFPI and extracellular DNA seems to be related to cancer process, not to local or systemic inflammation as these parameters differentiate cancer from chronic pancreatitis. Interestingly, D-dimers and TF-MP are related to the disseminated cancer stage, suggesting that vascular invasion is a prerequisite to the release of TF-MP from the primary tumor into the bloodstream and to coagulation activation. However, only D-dimers are associated with the occurrence of future VTE. We propose that D-dimers could be a useful tool to predict thrombotic events in pancreatic cancer patients. This should be confirmed in a larger population.

Identification and frequency of circulating CD4(+) T lymphocytes specific to Benzylpenicillin in healthy donors.

BACKGROUND: Drug hypersensitivity is known to rely on a drug-specific T-cell response. Amplitude of antigen-specific T-cell response is partly controlled by the size of the antigen-specific naïve CD4(+) T-cell repertoire, but estimate of this repertoire has never been investigated for allergenic drugs. The purpose of this study was to evaluate the frequency of benzylpenicillin-specific CD4(+) T lymphocytes in healthy donors. METHODS: Co-cultures were established with CD4(+) T lymphocytes from healthy donors and mature autologous dendritic cells loaded with benzylpenicillin coupled to human serum albumin. CD4(+) T lymphocytes were stimulated once a week for 4 weeks with benzylpenicillin coupled to human serum albumin. The CD4(+) T-cell response was measured using an interferon-γ ELISPOT assay. Frequency of benzylpenicillin-specific naive CD4(+) T lymphocytes was then calculated using the Poisson distribution law. RESULTS: Results showed the presence of benzylpenicillin-specific CD4(+) T lymphocytes in 9 of 10 tested healthy donors irrespective of their HLA typing, with a mean frequency of 0.29 cells per million of CD4(+) T cells. Experiments performed on naive (CD45RA(+) ) and on memory (CD45RO(+) ) CD4(+) T lymphocytes showed that these benzylpenicillin-specific CD4(+) T lymphocytes belonged to the naive T-cell subpopulation. CONCLUSION: This study showed for the first time the existence of naive CD4(+) T lymphocytes specific to benzylpenicillin in healthy donors.

Biological effect of ß-(1,3)-polyglucuronic acid sodium salt on lipid storage and adipocytes differentiation.

ß-(1,3)-Polyglucuronic acid sodium salt produced by the regioselective oxidation of ß-(1,3)-glucan have been studied for its biological impact as putative slimming agent. This ß-(1,3)-polyglucuronic acid sodium salt was synthesized using the conventional 2,2,6,6,-tetramethylpiperidine-1-oxyl radical (TEMPO)/NaBr/NaClO systems at pH 10 and 4°C. A transcriptomical study using DNA microarray analysis, demonstrated that this heparan like sodium salt locally induced an over-expression of the gene Angiopoietin-like 4 (ANGPTL4 or fasting induced adipose factor (FIAF)) leading to the increase in Adipokine ANGPTL4 synthesis and the inhibition of Lipoprotein Lipase (LPL). In vitro analysis using 3T3-L1 cells have clearly revealed that ß-(1,3)-polyglucuronic acid sodium salt could act in key steps of lipid metabolism by inhibiting the differentiation of pre-adipocytes to mature adipocytes.

Synthesis and application of a N-1' fluorescent biotinyl derivative inducing the specific carboxy-terminal dual labeling of a novel RhoB-selective scFv.

The fluorescent site-specific labeling of protein would provide a new, easy-to-use alternative to biochemical and immunochemical methods. We used an intein-mediated strategy for covalent labeling of the carboxy-terminal amino acid of a RhoB-selective scFv previously isolated from a phage display library (a human synthetic V(H) + V(L) scFv phage library). The scFv fused to the Mxe intein was produced in E. coli and purified and was then labeled with a newly synthesized fluorescent biotinyl cysteine derivative capable of inducing scFv-Mxe intein splicing. In this study, we investigated the splicing and labeling properties of various amino acids in the hinge domain between scFv and Mxe under thiol activation. In this dual labeling system, the fluorescein is used for antibody detection and biotin is used for purification, resulting in a high specific activity for fluorescence. We then checked that the purified biotinylated fluorescent scFv retained its selectivity for RhoB without modification of its affinity.

[Pharmacogenetics of anticancer drugs]. / Pharmacogénétique des médicaments anticancéreux.

Much progress has been made in treating human malignancies and there are now multiple treatment options with similar efficacy for nearly every type of cancer. However, the narrow therapeutic index of most chemotherapeutic agents and the severe consequences of undertreatment or overdosing have led to research molecular predictive factors of the toxicity and efficacy of cancer treatments. Genetic factors affecting drug metabolism and transport partly explain interindividual variability in drug response. Pharmacogenetic focuses on the molecular mechanisms involved in drug response, and its ultimate goal is the optimisation of the treatments, that combines the optimal efficacy and the minimal risk of severe side effects. Polymorphisms in genes encoding specific drug-metabolising enzymes can result in individuals in the general population being characterised as low, rapid or even ultra-rapid metabolisers. Phenotyping and genotyping tests are now available that determine or predict the metabolic status of an individual and, thus, enable the evaluation of risk of drug failure or toxicity. Some clinical applications of pharmacogenetics (5-FU, irinotecan, thiopurines) have already been developed in routine medicine resulting in significant improvement in patient treatment. The clinical validation of an increasing number of pharmacogenetic tests, as well as the development of new highly efficient technologies for genotyping (real-time PCR, DNA chips...) should further promote pharmacogenetics in clinical practice and lead to the development of a patient-tailored drug therapy.