[Impact of the suicidal crisis intervention training program on the confidence and skills of hospital professionals in the Hauts-de-France region]. / Impact de la formation à l'intervention de crise suicidaire sur la confiance et les aptitudes des professionnels hospitaliers des Hauts-de-France.

INTRODUCTION: Suicide is a major public health issue given its huge human and economic consequences. Symptoms prior to suicide are often not specific. Nevertheless, the majority of suicidal people express suicidal thoughts, and nearly one in two meet a health professional in the period preceding the act. Being able to recognize the warnings and intervene during the suicidal crisis, defined as a mental crisis where the major risk is suicide, is to seize the opportunity to postpone the suicidal plan and to gain time to implement in place lasting strategies to combat suffering. Thus, the training for suicidal crisis intervention is a major axis of the suicide prevention strategy. Recently, crisis intervention training programs have been updated with knowledge accumulated since the early 2000's. In France, one of the countries most concerned by suicide, the Hauts-de-France region is one of the most impacted. In this context, the Regional Health Agency of Hauts-de-France included in its Regional Health Program of 2018-2023 the training of healthcare workers who work with high suicidal risk patients. The suicidal crisis intervention training program (SCIT) has been introduced to hospital staffs in Hauts-de-France. The purpose of this study was to evaluate this program. METHODS: Eight training sessions with 15 to 21 participants were carried out from 2019 November to 2021 January in the Hauts-de-France region. Participants were volunteer healthcare professionals in direct contact with suicidal crisis patients. The training included three modules. The first one concerned the suicidal crisis intervention training: definition of the suicidal crisis, typology of the crisis, vulnerability development, crisis evaluation and crisis intervention practice. The second concerned the evaluation with the RED scale (Risk-Emergency-Danger) and the adequate patient orientation to a psychiatric unit. The third was dedicated to the Gatekeeper training with the constitution of a Gatekeeper network to enhance the capacity to detect suicidal risk and to orient the concerned person towards an adequate evaluation or care organization. We evaluated the first two levels of the Kirkpatrick's model: level 1) the participant's satisfaction (rated out of 10), and level 2) the degree of confidence in their professional abilities (rated out of 10) and their skills in responding to a person in a suicidal crisis (using the SIRI-2-VF - French version of the Suicide Intervention Response Inventory-2). The participants were interviewed before (T0), just after (T1) and at one month of training (T2). RESULTS: Among the 141 health professionals who followed the training, 139 answered the questionnaire at least one time (13 psychologists, 22 doctors, 97 nurses and 7 head nurses). The participation rates were 99.3 % at T0, 96.4 % at T1 and 46.0 % at T2. Most of the participants were nurses (69.8 %), and 33.1 % of the respondents declared they had already followed a suicidal crisis training. The satisfaction with the training was evaluated at 8.6 (± 1.3) out of 10. There was no significant difference among the professions, neither between those having already received or not a previous training. The self-perceived capacity to manage a suicidal crisis was rate 6.8 (± 1.8) out of 10 at T0. There was a significant increase just after the training (8.1±1.2 vs 6,8±1,8, p<0,001) which persisted at 1 month (8.1±1.1 vs 6.8±1.8, P<0.001). The score at the SIRI-2-VF was 15.0 (± 4.2) out of 30 at T0. There was a significant increase just after the training (17.5±3.5 vs 15.0±4.2, P<0.001), which persisted at 1 month (17.0±4.0 vs 15.0±4.2, P<0.001). DISCUSSION: This is the first evaluation of the suicidal crisis intervention training program. This program increased and homogenized the competency of the participants to manage suicidal ideation and behaviors. Those who followed a previous training maintained higher scores than the others, which shows the importance of repeated training to maintain a satisfying level of knowledge over the long term. One of the strengths of this training is the use of roleplay which enhances the learning and abilities to interact with people at suicidal risk. It seems important to integrate a suicidal crisis intervention training in the cursus of health students to avoid suicide and the dramatic consequences for the entourage and the health professionals who are confronted with it. CONCLUSION: The SCIT program showed encouraging results in terms of confidence and capacity of the healthcare professionals to intervene in suicidal crisis.

Memory and PTPIP51--a new protein in hippocampus and cerebellum.

Previously the expression of Protein Tyrosine Phosphatase Interacting Protein 51 (PTPIP51) in mouse brain was reported. Here, we investigated PTPIP51 mRNA and protein in two of the brain regions namely the hippocampus and the cerebellum of mouse brains. On a cellular level both the protein and the mRNA were related to the pyramidal cells of the hippocampal formation, the granular cells of the dentate gyrus and the cells of the adjacent strata. In the cerebellum PTPIP51 was traced in Purkinje cells, the cells of the molecular layer and the granular layer. On a subcellular level only partial co-localization was seen for the endoplasmic reticulum, but not with mitochondria. In addition the interactome of PTPIP51 was analysed. In hippocampal cells a strong interaction with PTP1B and vesicle-associated membrane protein-associated protein B (VAPB) was detected. A somewhat differing interaction profile was found in the cerebellum, where high interaction levels were found for 14-3-3, diacylglycerol kinase α (DGKα), NFκB and PTP1B. These interaction partners represent specific signalling pathways linked to building memory. PTPIP51 can be associated with nerve growth factor signalling, dendritic and axonal growth, synaptogenesis, and all processes needed for memory formation. Moreover, in HT-22 mouse hippocampal cells PTPIP51 expression was induced by administrating the fibroblast growth factor 1 (FGF-1), which is known to take part in learning/memory processes. Knocking down p38-MAPK also led to an up-regulation of PTPIP51 probably resembling a compensative mechanism. Thus, a possible connection to the processing of memories can be anticipated. Differences in the interaction profile in both regions may be attributed to the actual/local differences in memory formation.

PTPIP51: a new interaction partner of the insulin receptor and PKA in adipose tissue.

AIMS: Our previous experiments revealed an association of PTPIP51 (protein tyrosine phosphatase interacting protein 51) with the insulin signalling pathway through PTP1B and 14-3-3beta. We aimed to clarify the role of PTPIP51 in adipocyte metabolism. METHODS: Four groups of ten C57Bl/6 mice each were used. Two groups were fed a standard diet; two groups were fed a high-fat diet. Two groups (one high-fat diet and one standard diet) were submitted to endurance training, while the remaining two groups served as untrained control groups. After ten weeks, we measured glucose tolerance of the mice. Adipose tissue samples were analyzed by immunofluorescence and Duolink proximity ligation assay to quantify interactions of PTPIP51 with either insulin receptor (IR) or PKA. RESULTS: PTPIP51 and the IR and PTPIP51 and PKA, respectively, were colocalized in all groups. Standard diet animals that were submitted to endurance training showed low PTPIP51-IR and PTPIP51-PKA interactions. The interaction levels of both the IR and PKA differed between the feeding and training groups. CONCLUSION: PTPIP51 might serve as a linking protein in adipocyte metabolism by connecting the IR-triggered lipogenesis with the PKA-dependent lipolysis. PTPIP51 interacts with both proteins, therefore being a potential gateway for the cooperation of both pathways.

PTPIP51, a positive modulator of the MAPK/Erk pathway, is upregulated in glioblastoma and interacts with 14-3-3ß and PTP1B in situ.

Glioblastoma multiforme (GBM) is the most common and most malignant primary brain tumour. Protein tyrosine phosphatase interacting protein 51 (PTPIP51) is an interaction partner of 14-3-3ß, which correlates with the grade of malignancy in gliomas. In this study PTPIP51 and its interacting partners 14-3-3ß, PTP1B, c-Src, Raf-1 as well as EGFR were investigated in human glioblastoma. Twenty glioblastoma samples were analyzed on transcriptional and translational level by immunohistochemistry, in situ hybridization and RT-PCR. To compare PTPIP51 expression in gliomas of different malignancies, quantitative RT-PCR for grade II astrocytoma and GBM samples was employed. Additionally, we analyzed the correlation between PTPIP51 and 14-3-3ß transcription, and checked for in situ interaction between PTPIP51 and 14-3-3ß and PTP1B, respectively. PTPIP51 and 14-3-3ß mRNA showed a tumour grade dependent upregulation in gliomas. Glioblastoma cells displayed a strong immunoreaction of PTPIP51, which co-localized with 14-3-3ß and PTP1B. The duolink proximity ligation assay corroborated a direct in situ interaction of PTPIP51 with both proteins, known to interact with PTPIP51 in vitro. The in vitro interacting partners Raf-1 and c-Src showed a partial co-localization. Besides, immune cells located in capillaries or infiltrating the tumour tissue and endothelial cells of pseudoglomerular vessels revealed a high PTPIP51 expression. The upregulation of PTPIP51 and its connection with the EGFR/MAPK pathway by 14-3-3ß via Raf-1 and by PTP1B via c-Src, argue for a functional role of PTPIP51 in the pathogenesis of human glioblastoma.

Expression profile of PTPIP51 in mouse brain.

This study demonstrates the expression of the novel protein protein tyrosine phophatase-interacting protein 51 (PTPIP51) in mammalian brain tissue. Serial sections of the whole adult mouse brain were analyzed for PTPIP51 protein and mRNA by immunohistochemistry, immunoblotting, RT-PCR, and in situ hybridization. Recent investigations by Yu et al. (2008) describe PTPIP51 as being capable of activating Raf-1, thereby modulating the MAPK pathway. The role of Raf-1, as well as of 14-3-3, in neurological disorders is well established. PTPIP51 expression was confined to neurons in the following structures: the piriform cortex and their connections to the anterior commissure, nucleus accumbens, paraventricular and supraoptical nuclei, neurohypophysis, superior colliculus, genu of facialis nerve, spinal trigeminal tract, inferior cerebellar peduncle, and cerebellum. In the cerebellum, a subpopulation of Purkinje cells and their dendrites was strongly PTPIP51 positive. Moreover, PTPIP51 was found to be colocalized with vasopressin and its transport protein neurophysin II in the neuroendocrine nuclei and their connections to the neurohypophysis. The data presented here suggest a role of PTPIP51 in neuronal homeostasis, axonal growth, and transport.

The novel protein PTPIP51 is expressed in human keratinocyte carcinomas and their surrounding stroma.

BACKGROUND: The novel protein PTPIP51 (SwissProt accession code Q96SD6) is known to interact with two non-transmembrane protein-tyrosine phosphatases, PTP1B and TCPTP in vitro. Overexpression of the full-length protein induces apoptosis in HeLa and HEK293T cells (Lv et al. 2006). PTPIP51 shows a tissue-specific expression pattern and is associated with cellular differentiation and apoptosis in some mammalian tissues, especially in human follicular and interfollicular epidermis. PTPIP51 protein is expressed in all suprabasal layers of normal epidermis, whereas the basal layer contains PTPIP51 mRNA only but lacks the protein. OBJECTIVES: The expression of PTPIP51 was investigated in keratinocyte carcinomas, that is human basal cell carcinomas (BCCs) and squamous cell carcinomas (SCCs) as well as Bowen's disease (BD) and keratoacanthomas (KAs) on a transcriptional (mRNA) and translational (immunohistochemical) level. METHODS: Formalin-fixed, paraffin-embedded sections of BCCs, SCCs, KAs and BD, respectively, were analysed by RT-PCR, as well as immunohistochemistry and subsequent fluorescence microscopy. PTPIP51-positive cells of the tumour and the surrounding stroma were identified on the basis of specific morphological features by means of H & E staining. To obtain further information about a putative function of PTPIP51, a possible association of PTPIP51 with apoptotic cells, as well as an assumed negative correlation with proliferating cells was investigated by means of an in-situ TUNEL assay and Ki67/MIB-1 antigen staining, respectively. Co-immunostainings with PTPIP51 were performed for the following antigens: TCPTP, PTP1B and beta-catenin. RESULTS: PTPIP51-expression was detected in BCCs and SCCs of the skin, as well as in KAs and BD. Both types of keratinocyte carcinoma revealed a specific localization pattern of PTPIP51 in malignant keratinocytes. Whereas PTPIP51-positive cells of BCC were found to form two cluster types with a different subcellular localization of the protein, i.e. cytoplasmic and nuclear or predominantly membranous, investigation of SCC revealed a meshwork-like appearance of PTPIP51-positive malignant keratinocytes, created by a mainly membranous localization. BD and KA resembled the findings of PTPIP51-expression in SCC. Furthermore, we observed a partial co-localization of PTP1B and PTPIP51 in BCC. SCC and BCC showed a co-expression and partial co-localization of PTPIP51 with beta-catenin. Some PTPIP51-positive cells were found to undergo apoptosis. PTPIP51 was also expressed in cells comprising the surrounding stromal microenvironment. This was particularly noticed for endothelial cells lining peritumoural vessels as well as for infiltrating cells of both, the innate and the adaptive immune system. CONCLUSIONS: The results showed a distinct mainly membranous expression pattern of PTPIP51 in BCCs and SCCs. Since PTPIP51 was also detected in the peritumoural tissue, the protein may play a crucial role in ke

Origin of laurdan sensitivity to the vesicle-to-micelle transition of phospholipid-octylglucoside system: a time-resolved fluorescence study.

The fluorescent probe laurdan has been shown to be sensitive to the vesicle-to-micelle transition of phosphatidylcholine/octylglucoside (M. Paternostre, O. Meyer, C. Grabielle-Madelmont, S. Lesieur, and, Biophys. J. 69:2476-2488). On the other hand, a study on the photophysics of laurdan in organic solvents has shown that the complex de-excitation pathway of the probe can be described by two successive processes, i.e., an intramolecular charge transfer followed by dielectric relaxation of the solvent if polar. These two excited-state reactions lead to three emitting states, i.e., a locally excited state, a charge transfer state, and a solvent relaxed state (M. Viard, J. Gallay, M. Vincent, B. Robert and, Biophys. J. 73:2221-2234). Experiments have been performed using time-resolved fluorescence on the probe inserted in amphiphile aggregates (mixed liposomes, mixed micelles) different in detergent-to-lipid ratios. The results have been compared with those obtained for laurdan inserted in dipalmitoyl phosphatidylcholine liposomes in the gel and in the fluid lamellar phase. Except for laurdan in dipalmitoyl phosphatidylcholine liposomes in the gel lamellar phase, the red part of the emission spectra originates from the de-excitation of the relaxed excited state of laurdan, indicating that indeed the dielectric relaxation process is an important phenomena in the ground-state return pathway of this probe. On the other hand, the maximization entropy method (MEM) analysis of the fluorescence decay recorded in the blue part of the emission spectra indicates that the dielectric relaxation is not the only reaction occurring to the excited state of laurdan. Moreover, the analysis of the fluorescence decays of laurdan inserted in gel lamellar dipalmitoylphosphatidylcholine (DPPC) liposomes indicates excited-state reactions, although dielectric relaxation is impossible. These results are in agreement with the de-excitation pathway determined from laurdan behavior in organic solvent even if, in most of the aggregates studied in this work, the major phenomenon is the dielectric relaxation of the solvent. All along the vesicle-to-micelle transition, we have observed that the lifetime of the relaxed excited state of laurdan continuously decreases probably due to a dynamic quenching process by water molecules. On the other hand, the time constant of the dielectric relaxation process remains almost unchanged in the lamellar part of the transition but abruptly decreases as soon as the first mixed micelle is formed. This decrease is continuous all over the rest of the transition even if it is more pronounced in the mixed liposomes' and mixed micelles' coexistence. The increase of the octylglucoside-to-lipid ratio of the mixed micelles via the change of the size and the shape of the aggregates may facilitate the penetration and the mobility of water molecules. Therefore, during the vesicle-to-micelle transition, laurdan probes the evolution of both the amphiphile packing in the aggregates and the increase of the interface polarity. This study finally shows that the detergent-to-lipid ratio of the mixed micelles is an important parameter to control to limit the penetration and the mobility of water within the amphiphile aggregates and that laurdan is a nice tool to monitor this phenomenon.

Laurdan solvatochromism: solvent dielectric relaxation and intramolecular excited-state reaction.

Absorption, steady-state, and time-resolved fluorescence measurements have been performed on laurdan dissolved either in white viscous apolar solvents or in ethanol as a function of temperature. The heterogeneity of the absorption spectra in white oils or in ethanol is consistent with semiempirical calculations performed previously on Prodan. From steady-state and time-resolved fluorescence measurements in apolar media, an excited state reaction is evidenced. The bimodal lifetime distribution determined from the maximum entropy method (MEM) analysis is attributed to the radiative deexcitation of a "locally excited" (LE) state and of a "charge transfer" (CT) state, whereas a very short component (20 ps), the sign and the amplitude of which depend on the emission wavelength, is attributed to the kinetics of the interconvertion reaction. The observation of an isoemissive point in the temperature range from -50 degrees C to -110 degrees C in ethanol suggests an interconvertion between two average excited-state populations: unrelaxed and solvent-relaxed CT states. A further decrease in temperature (-190 degrees C), leading to frozen ethanol, induces an additional and important blue shift. This low temperature spectrum is partly attributed to the radiative deexcitation of the LE state. Time-resolved emission spectra (TRES) measurements at -80 degrees C in the ethanol liquid phase show a large spectral shift of approximately 2500 cm(-1) (stabilization energy of the excited state: 7.1 kcal x M(-1)). The time-dependent fluorescence shift (TDFS) is described for its major part by a nanosecond time constant. The initial part of the spectral shift reveals, however, a subnanosecond process that can be due to fast internal solvent reorientation and/or to intramolecular excited-state reactions. These two relaxation times are also detected in the analysis of the fluorescence decays in the middle range of emission energy. The activation energy of the longest process is approximately 3 kcal x M(-1). At -190 degrees C, one subnanosecond and one nanosecond excited-state reactions are also evidenced. They are likely due to intramolecular rearrangements after the excitation, leading to the CT state and not to solvent relaxation, which is severely hindered in these temperature conditions. Therefore, both intramolecular and solvent relaxations are responsible for the large Stokes shift displayed by this probe as a function of solvent polarity. A possible scheme is proposed for the deexcitation pathway, taking into account the kinetics observed in these different solvents.

Solubilization and reconstitution of vesicular stomatitis virus envelope using octylglucoside.

Reconstituted vesicular stomatitis virus envelopes or virosomes are formed by detergent removal from solubilized intact virus. We have monitored the solubilization process of the intact vesicular stomatitis virus by the nonionic surfactant octylglucoside at various initial virus concentrations by employing turbidity measurements. This allowed us to determine the phase boundaries between the membrane and the mixed micelles domains. We have also characterized the lipid and protein content of the solubilized material and of the reconstituted envelope. Both G and M proteins and all of the lipids of the envelope were extracted by octylglucoside and recovered in the reconstituted envelope. Fusion activity of the virosomes tested either on Vero cells or on liposomes showed kinetics and pH dependence similar to those of the intact virus.

Protein Phosphorylation Is Induced in Tobacco Cells by the Elicitor Cryptogein.

Changes in plasmalemma ion fluxes were observed when tobacco (Nicotiana tabacum) cells were treated with cryptogein, a proteinaceous elicitor from Phytophthora cryptogea. A strong alkalization of the culture medium, accompanied by a leakage of potassium, was induced within a few minutes of treatment. These effects reached a maximum after 30 to 40 min and lasted for several hours. This treatment also resulted in a rapid, but transient, production of activated oxygen species. All these physiological responses were fully sensitive to staurosporine, a known protein kinase inhibitor. Furthermore, a study of protein phosphorylation showed that cryptogein induced a staurosporine-sensitive phosphorylation of several polypeptides. These data suggest that phosphorylated proteins may be essential for the transduction of elicitor signals.