Somatic intragenic recombination of the arylsulfatase A gene in a metachromatic leukodystrophy patient

A metachromatic leukodystrophy (MLD) patient was found to carry two additional arylsulfatase A (ARSA) alleles besides the two inherited. The additional alleles arose from an event of mitotic intragenic recombination between the inherited alleles, thus leading to a case of somatic mosaicism. As suggested by in vitro expression, the recombination was ineffective in generating a significantly advantaged ARSA allele compared to the inherited alleles. Although the phenotype in this patient was not modified by the recombination, similar events could potentially yield significant clinical benefits.     

The role of voluntary exercise in enriched rearing: a behavioral analysis

The effects of postweaning enriched rearing and home cage voluntary wheel-running exercise in adulthood were contrasted on a comprehensive battery of tests designed to assess mnemonic, attentional, emotional, and motor functions. In a 2 x 2 factorial design, female C57BL/6 mice were housed in groups in either standard or enriched cages, which were equipped with either a running or a locked wheel. They were maintained in the corresponding housing conditions for 2 months postweaning prior to, and throughout, testing. Enriched rearing was associated with anxiogenesis, hypolocomotor activity, enhanced motor skills, retarded extinction of conditioned responding, and improved water maze performance. Exercise as such enhanced motor coordination and facilitated extinction of contextual conditioning. Evidence for an interaction between enrichment and exercise was apparent in the open field test, conditioned freezing to a tone stimulus, prepulse inhibition, and acquisition of water maze reference memory. Hence, care should be taken to control for the unique contribution of wheel-running exercise when it is included as an integral component of the enrichment procedure.     

Decrease of current responses at human recombinant P2X3 receptors after substitution by Asp of Ser/Thr residues in protein kinase C phosphorylation sites of their ecto-domains

The whole-cell patch-clamp technique was used to record current responses to nucleotides in HEK 293 cells transiently transfected with the human (h) P2X(3) receptor. When GDP-beta-S was included into the pipette solution, UTP at concentrations which did not alter the holding current, facilitated the alpha,beta-methylene ATP (alpha,beta-meATP)-induced current. The substitution of Ser/Thr residues situated within protein kinase C (PKC) consensus phosphorylation sites of the P2X(3) receptor ecto-domain by the neutral amino acid Ala either abolished (T134A, S178A) or did not alter (T196A, S269A) the UTP-induced potentiation of the alpha,beta-meATP current. The substitution of the same Ser/Thr residues in all four PKC sites by the negatively charged Asp prevented the potentiation by UTP. The Asp mutations abolished the first, fast offset time-constant, but did not alter, or in the case of S269D even increased, the second, slow offset time-constant; at the same time such mutations invariably increased the onset time-constant and massively depressed the peak current amplitude. None of the Ala mutations (with the exception of S269A) influenced the time-course of desensitisation or the peak current amplitude. It is concluded that constitutive activation of PKC sites at the ecto-domain of the hP2X(3) receptor both abolishes the UTP-induced potentiation of the alpha,beta-meATP current and accelerates its rate of desensitisation.     

Cerebellar neurometabolite abnormalities in pediatric attention/deficit hyperactivity disorder: A proton MR spectroscopic study

We designed a case–control proton magnetic resonance spectroscopic study comparing the cerebellar and prefrontal regions of a group of 17 ADHD (attention deficit/hyperactivity disorder) medicated children and a group of 17 control children matched for laterality, gender and age. As we had found decreased gray matter volume in the right prefrontal region and the left cerebellar hemisphere in a previous voxel-based morphometry study conducted on an independent ADHD sample, we tested the hypothesis that these regions should show neurometabolite abnormalities. MRI (magnetic resonance imaging) was performed with a 1.5 T system; spectral acquisition was performed with a single-voxel technique and a PRESS sequence. Two volumes of interest were selected in the right prefrontal region and the left cerebellar hemisphere. NAA (N-acetylaspartate), Cre (creatine), Cho (choline), MI (myo-inositol) and Glx (glutamate-glutamine) resonance intensities were absolutely quantified. In the left cerebellar hemisphere, ADHD children showed significant decreased MI and NAA absolute concentrations with high effect sizes (p = 0.004, ES = 1.184; p = 0.001, ES = 1.083). The diminished absolute concentration of the NAA could be related to a gray matter volume decrease in the same cerebellar region found in the previous voxel-based morphometry MRI study, while the reduced MI absolute concentration could express a decreased glial density. This is the first proton MR spectroscopic study examining the cerebellum and it provides additional support for the role of cerebellum in the ADHD neurobiology.     

Professional competencies learned through working on a medication education project

Objectives

To implement a medication education project and assess the competencies students learned and implemented in professional practice after graduation.

Design

Fourth-year pharmacy students planned, carried out, and reported on a real-life project during 1 study year. Outside experts and 2 faculty members facilitated the work. The aim of the medication education project was to create material that schoolteachers could use to teach children about rational use of medicines.

Assessment

All students who had participated in the medication education program during its 3 years were contacted (n = 31). A questionnaire was sent to the 21 students who had graduated (18 responded), and a focus group was conducted with the 10 students completing their final year of pharmacy school (9 participants). The competencies that the students reported learning most were teamwork and social interaction skills. They considered the project motivating but also found it challenging and the deadlines frustrating.

Conclusions

Through participation in a medication education project, students learned interpersonal skills, time management, conflict resolution, and other skills that many of them already were finding valuable in their professional practice.     

Anti-inflammatory and analgesic effects displayed by peptides derived from PKI55 protein, an endogenous protein kinase C inhibitor

We recently characterized the PKI55 protein as an endogenous protein kinase C (PKC) inhibitor and investigated, in vitro, the potential anti-inflammatory actions of its N-terminal peptides 1–16 (peptide 5), 1–8 (peptide 8) and 1–5 (peptide 9). We showed their ability to inhibit chemotaxis in human polymorphonuclear leukocytes activated by the N-formyl tripeptide for-Met-Leu-Phe-OMe. In this work, we evaluated the anti-inflammatory and the analgesic effects of the selected peptides by in vivo experiments carried out in the mouse. The peptides 5, 8 and 9 (0.1 and 10 nmol i.c.v.) were effective in both the parameters chosen to test the anti-inflammatory activity, i.e., the xylene-induced ear edema and the acetic acid-induced infiltration of neutrophils in the peritoneal cavity. In addition, they displayed analgesic effect, evaluated by the acetic acid-induced writhing test. All the peptides’ effects were shared by the reference compounds, dexamethasone and indomethacin (10 mg?kg^?1?i.p.), but not by the 9-scramble peptide (10 nmol i.c.v.). The peptide 9, which represents the shortest active sequence of the PKI55 protein, was tested in the ear edema model even following intraperitoneal (i.p.) administration and proved to be effective in the range doses 3–30 mg?kg^?1. Moreover, an increase in plasma corticosterone levels was detected in mice treated with the peptide 9, but not with the 9-scramble peptide (both at 10 nmol i.c.v.). The anti-inflammatory and analgesic effects of the PKI55-derived synthetic peptides, possibly related both to PKC inhibition and hypothalamic–pituitary–adrenal axis activation, deserve further investigation in view of potential therapeutic exploitation.