The use of transient gene expression assays for the study of natural or engineered plant promoters is affected by a considerable degree of inter-experiment variability. As a means of obtaining interpretable data from a limited number of experiments, we worked out conditions for the simultaneous determi nation of the activity of two reporter genes, a "sample" and a "reference", ona single extract of co-transformed protoplasts. ß-glucuronidase (GUS) and chloramphenicol acetyl transferase (CAT) genes, both under the control of the CaMV 35S promoter, were transferred into tobacco (Nicotiana tabacum L.) protoplasts on two independent plasmids. The parallel expression of the two reporter genes in several independent co-transformation experiments was verified. Conditions for the use of a single protoplast extraction buffer and for the simultaneous assay of both reporter gene activities were set up. A HPLC method for the non-radioactive determination of both enzyme activities on a single aliquot of the reaction mixture was developed. The resulting procedure was tested using the GUS gene as "reference" and the CAT gene, under the control of either wild type or upstream-deleted (-90) CaMV 35S promoter, as "sample". The protocol is simple and allows the fast analysis of plant promoters in the presence of a true internal standard under conditions in which assay manipulations are reduced to a minimum and both reporter gene activities are subjected to the same experimental treatments.

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IntroductionSelf Disorders (SDs) are regarded as the subjective phenotype of Schizophrenia vulnerability. The EASE (Examination of Anomalous Self-Experiences) scale is the most detailed and widely used instrument to investigate SDs, but it requires long administration times and specific training. The IPASE (Inventory of Psychotic-like Anomalous Self-Experiences) scale might be a self-administered instrument of widespread use for an easier SDs investigation.ObjectivesThe present study was aimed at validating the Italian version of IPASE, testing its internal consistency and usability for a first level SDs survey. A secondary objective was to confirm the correlations between IPASE, EASE, main symptom dimensions, subjective bodily experiences, symptoms of schizophrenic autism as well as levels of global functioning.MethodsFifty patients with Schizophrenia were administered the IPASE scale in its Italian version, the Examination of Anomalous Self-Experiences scale (EASE), the Positive And Negative Symptoms Scale (PANSS), the Social and Occupational Functioning Assessment Scale (SOFAS) to assess global functioning, the Autism Rating Scale (ARS) and the Abnormal Bodily Phenomena questionnaire (ABPq). The internal consistency of IPASE in its Italian version was investigated and the correlations between IPASE, EASE, ABP, ARS, PANSS and SOFAS were explored.ResultsThe internal consistency of the Italian version of IPASE was high (α 0.97). The IPASE and EASE total scores were positively correlated with each other, as were many of the conceptually related subdomains of both scales. The IPASE score was negatively correlated with global functioning (SOFAS) and positively correlated with total PANSS scores and with PANSS negative domain. Moreover, the IPASE total score was positively correlated with autism dimension (ARS), while anomalies in subjective experience of the lived body were coherently correlated with higher scores in IPASE “somatization” subdomain.Conclusions The IPASE may be an easy instrument with high internal consistency for an initial investigation of SDs. IPASE domains appear to be correlated with the SDs investigated through EASE and with the main symptomatologic dimensions of Schizophrenia, in particular with negative symptoms. IPASE might also be a useful instrument for a first level investigation of subjective experiences concerning intersubjectivity and bodily dimensions.SDs are confirmed to be a core feature of the schizophrenia psychopathology, with a adverse impact on global functioning.Disclosure of InterestNone Declared

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