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Title: The impact of Clown Therapy in a Palliative Care Unit (PCU); a pilot studyBackground: Although several studies on clown therapy have demonstrated benefits for children and the elderly, few studies exist on its impact on quality of life in adult terminal patients.We devised this monocentric, prospective, descriptive pilot study, covering both quantitative and qualitative aspects, to test the hypothesis that “clowning” in the PCU would help to improve the quality of life of patients, and benefit those accompanying them and their carers.Methods: 30 terminal patients [age range 63-79, 19 women, 11 men] from the palliative care unit at Nîmes University Hospital were included in the study from 31/03/2016 to 08/11/2016. All had given written informed consent. 2 clowns visited the patients and their companions, inviting them to take part in 15 to 30-minute improvised role-plays to prepare them for the inevitable separation. Primary outcome measures for the impact of the clowns’ interventions were: The Edmonton Symptom Assessment System evaluating patients’ physical symptoms and the McGill Quality of Life Questionnaire assessing patients’ quality of life. The secondary outcome measure was Alceste textual data analysis software to analyse semi-structured interviews held with patients, their companions and carers. Results: However long the clown’s intervention, the ESAS scores indicated a significant improvement in nausea (p=0.0248) and dyspnea (p=0.0476). Scores for depression (p=0.0160) also showed an improvement. A slight difference in the scores for anxiety (p=0.0900) and drowsiness (p=0.0812) was also observed. 2 days after the clowns’ intervention, the Mc Gill scores revealed a significantly greater physical quality of life (p=0.0044) and a very slight increase in the patients’ overall quality of life (p=0.0807). Results of the lexical analysis with Alceste revealed benefits for all those involved and gave an indication for better patient management at the PCU. Conclusions: This pilot study demonstrating the impact of “clowning” on various physical and mental symptoms in adult patients indicates the benefits of such interventions for patients, their companions and carers. Although the usefulness of clowns for children with life-threatening diseases is clear, the results obtained with our study clearly indicate the benefits of these interventions in adult, end-of-life patients. Further studies would be required to confirm these findings.Date of first registration : 20/11/2015Trial registration: ClinicalTrials.gov ID: NCT02610452This research was registered under the following reference : IDRCB 2015-A00719-40 and approved by the local committee for the protection of persons, CPP Sud-Méditerranée III-Nîmes on November 6th, 2015 under the reference 2015.06.03.

Reactive gliosis, leading to the formation of glial scar, is the response of astrocytes to CNS injury. It is an ubiquitous reaction observed in a large number of pathological conditions, such as mechanical and chemical lesions, as well as degenerative processes (Fulcrand and Privat, 1977; Eng and De Armond, 1982). This reaction is twofold, being characterized by an astrocyte multiplication (hyperplasia) as well as an hypertrophy of the perikarya and processes (Hain et al., 1960; Nathaniel and Nathaniel, 1981). The main intracellular event is an increase of the number of gliafilaments, paralleled by a raise in GFAP immunoreactivity.

The expression of glial fibrillary acidic protein (GFAP)-mRNA during mouse brain development and in astroglial primary cultures has been investigated by using two approaches: Northern-blot evaluation using a specific cDNA probe, and cell-free translation associated with immunoprecipitation. During brain maturation (4-56 days postnatal), the GFAP-mRNA underwent a biphasic evolution. An increase was observed between birth and day 15 (i.e., during the period of astroglial proliferation), which was followed by a decrease until day 56 (i.e., during astroglial cell differentiation). At older stages (300 days), an increase was observed, which might reflect gliosis. During astroglial in vitro development (7-32 days in culture), the GFAP-mRNA showed similar variations. An increase, observed during the period of astroglial proliferation (7-18 days), was followed by a decrease which occurred in parallel to marked changes in cell shape, cell process outgrowth, and the organization and accumulation of gliofilaments. During the same culture period (7-32 days), alpha-tubulin mRNA, which was used as an internal standard, did not vary significantly. These results show that the increase of the GFAP protein and of gliofilaments observed both in vivo and in vitro during astroglial differentiation cannot be ascribed to an accumulation of the GFAP-mRNA. It might be that more than one mechanism regulates the levels of free and polymerized GFAP and of its encoding mRNA.

The time course changes in levels of mRNA encoding glutamic acid decarboxylase (GAD) and proenkephalin (PPE) was analyzed in the rat striatum following unilateral lesion of substantia nigra with 6-hydroxydopamine. The levels of both GAD and PPE mRNAs increased after the dopaminergic deafferentation, reaching concomitantly a maximal twofold increase on day 25. Thereafter, the mRNA levels declined; at 4 months, the amount of PPE mRNA remained slightly elevated whereas GAD mRNA had returned to the control value, suggesting the action of a compensatory mechanism. We also observed a rise of glial fibrillary acidic protein mRNA level which reflects a reactive astrocytosis. In contrast, alpha-tubulin mRNA level remained unchanged, indicating that no significant synaptogenesis occurs in this experimental situation. No obvious modification in mRNA levels was detected in the striatum contralateral to the lesion. These results highlight the role of the modulation of gene expression in adaptive processes to dopamine deficiency in striatal efferent pathways. Its relevance to the pathophysiology of Parkinson's disease is discussed.

Sweet clover leaves accumulate o-coumaric acid glucoside in the vacuole of the mesophyll cells. Protoplasts isolated from these cells are able to synthesize o-coumaric acid glucoside from exogeneous [14C]phenylalanine. Using this simplified system, kinetic analyses have shown that the glucoside is synthesized in the cytoplasm and thereafter transferred into the vacuole. No further degradation of the glucoside occurs in the isolated protoplasts after this transfer. Under our conditions, the vacuolar pH is reduced by light/dark transition and by inhibitors of photosynthesis. These treatments also depressed synthesis and transport of o-coumaric acid glucoside. The data support a relationship between the functioning of photosynthesis and pH cytoplasm/vacuole-dependent transport of o-coumaric acid glucoside into the vacuole. However as o-[ 14 C ] coumaric acid glucoside is taken up by isolated vacuoles without any energy requirement, no final conclusions can be drawn as to the coupling of o-coumaric acid glucoside transport with the acidification process. In intact plant or isolated protoplasts o-coumaric acid glucoside is synthesized as the trans isomer but accumulated in the vacuole in the cis form. When loaded with trans o-[ 14 C ] coumaric acid glucoside, reconstituted tonoplastic vesicles released this compound during molecular sieving on a Sepharose 4B column. In contrast, the labelled cis form was completely retained inside the vesicular lumen in a parallel experiment. These results suggest a selective permeability of the tonoplast to the two isomers which could be involved, in vivo, in the trapping of the glucoside inside the vacuole and its irreversible accumulation.

International audience; GFAP mRNA levels were quantified by Northern blot analysis using a human GFAP (glial fibrillary acidic protein) cDNA probe in association with immunocytochemistry. Ten days after a unilateral lesion of substantia nigra with 6-hydroxydopamine (6-OHDA), GFAP mRNA level is increased 1.4-fold in the ipsilateral striatum; thereafter it declined continuously to reach the control level 4 months later. This effect contrasted with the lower and more sustained increase of preproenkephalin (PPE) mRNA, a marker of neuronal target of nigrostriatal pathway. Following ibotenic acid-induced neuronal degeneration of the neostriatum in the rat, we observed a sharp elevation of the GFAP transcripts (4-fold) as soon as 2 days after the lesion both in the striatum and in the substantia nigra. Whereas in the striatum GFAP mRNA level already declined at 5 days postlesion, it remained stable in the substantia nigra. In comparison GFAP immunoreactivity was slightly delayed. No obvious modification was observed in the contralateral side to the lesion whatever the denervation condition studied. Implication of these results on the understanding and the therapeutic approach of glial scarring is discussed.