Abstract Pompe disease, an autosomal recessive lysosomal storage disorder, is caused by deficiency of lysosomal acid alpha‐glucosidase (GAA). On cellular level, there is lysosomal‐bound and free accumulation of glycogen and subsequent damage of organelles and organs. The most severe affected tissues are skeletal muscles and heart. The only available treatment to date is an enzyme replacement therapy (ERT) with alglucosidase alfa, a recombinant human GAA (rhGAA) modified with mannose‐6‐phosphate (M6P), which is internalized via M6P‐mediated endocytosis. There is an unmet need to improve this type of therapy, especially in regard to skeletal muscle. Using different tissue culture models, we recently provided evidence that a moss‐derived nonphosphorylated rhGAA (moss‐GAA), carrying a glycosylation with terminal N‐acetylglucosamine residues (GnGn), might have the potential to improve targeting of skeletal muscle. Now, we present a pilot treatment of Gaa−/− mice with moss‐GAA. We investigated general effects as well as the uptake into different organs following short‐term treatment. Our results do confirm that moss‐GAA reaches the target disease organs and thus might have the potential to be an alternative or complementary ERT to the existing one.

Pompe disease is an autosomal recessive lysosomal storage disorder (LSD) caused by deficiency of lysosomal acid alpha-glucosidase (GAA). The result of the GAA deficiency is a ubiquitous lysosomal and non-lysosomal accumulation of glycogen. The most affected tissues are heart, skeletal muscle, liver, and the nervous system. Replacement therapy with the currently approved enzyme relies on M6P-mediated endocytosis. However, therapeutic outcomes still leave room for improvement, especially with regard to skeletal muscles. We tested the uptake, activity, and effect on glucose metabolism of a non-phosphorylated recombinant human GAA produced in moss (moss-GAA). Three variants of moss-GAA differing in glycosylation pattern have been analyzed: two with terminal mannose residues in a paucimannosidic (Man3) or high-mannose (Man 5) configuration and one with terminal N-acetylglucosamine residues (GnGn). Compared to alglucosidase alfa the moss-GAA GnGn variant showed increased uptake in differentiated myotubes. Moreover, incubation of immortalized muscle cells of Gaa−/− mice with moss-GAA GnGn led to similarly efficient clearance of accumulated glycogen as with alglucosidase alfa. These initial data suggest that M6P-residues might not always be necessary for the cellular uptake in enzyme replacement therapy (ERT) and indicate the potential of moss-GAA GnGn as novel alternative drug for targeting skeletal muscle in Pompe patients.

Host cell proteins are inevitable contaminants of biopharmaceuticals. Here, we performed detailed analyses of the host cell proteome of moss ( Physcomitrella patens) bioreactor supernatants using mass spectrometry and subsequent bioinformatics analysis. Distinguishing between the apparent secretome and intracellular contaminants, a complex extracellular proteolytic network including subtilisin-like proteases, metallo-proteases, and aspartic proteases was identified. Knockout of a subtilisin-like protease affected the overall extracellular proteolytic activity. Besides proteases, also secreted protease-inhibiting proteins such as serpins were identified. Further, we confirmed predicted cleavage sites of 40 endogenous signal peptides employing an N-terminomics approach. The present data provide novel aspects to optimize both product stability of recombinant biopharmaceuticals as well as their maturation along the secretory pathway. Data are available via ProteomeXchange with identifier PXD009517.

A survey was conducted in a South London CAMHS before and after the publication and implementation of NICE guidelines of 2005 for the treatment of depression in children and adolescents. The results for 2006 indicate that 28% of cases were receiving medication without psychological therapy. Of those prescribed medication, 96% were receiving a prescription for fluoxetine. Of those receiving psychological therapy most received cognitive behaviour therapy whilst none received interpersonal therapy. Although 72% of cases were receiving medication with psychological therapy this falls short of the 100% expectation of the NICE guidelines. Other services are encouraged to survey young people receiving ant-depressant medication against the NICE guidelines.

Background: Psychotropic medication is being prescribed with increasing frequency for children and adolescents but there are very few prescription reviews on the extent of use in the UK.Method: A retrospective prescriptions’ survey was carried out in an outpatient CAMHS in three time‐bands between 1997 and 2002.Results: We found a five‐fold increase in the number of prescriptions, a ten‐fold increase in the number of young people prescribed medications, and a dramatic increase in the range of psychotropics prescribed.Conclusions: There is a need to further evaluate prescribing for children and adolescents in the UK. Clearer guidelines should be developed to ensure safe practices.

Plant expression systems offer a valuable alternative to traditional systems for the production of recombinant biopharmaceuticals. A highly efficient polyethyleneglycol (PEG)-mediated transient expression system for secreted recombinant proteins in plants has been developed. The human vascular endothelial growth factor 121 (rhVEGF) has been successfully expressed and efficiently secreted into the culture medium by transiently transformed moss protoplasts. In order to obtain secretion efficiency data, different expressed signal peptides were analysed and time course studies were performed with expression constructs containing different promoters. The transformation procedure was optimised for high level expression (up to 10 microg/ml) and successfully performed even with a transgenic glyco-engineered strain lacking plant-specific immunogenic sugar residues in N-glycans. The amount of rhVEGF was produced in such quantity that it allowed for the analysis of biological activity, silver-staining and Western blotting, revealing the correct formation and processing of the homodimer. This fast and flexible transient expression system enables feasibility studies and construct optimisation to be concluded within a few days, thus avoiding the time consuming step of having to generate stably transformed lines.

Child and Adolescent Mental HealthVolume 9, Issue 4 p. 199-199 Social Aggression Among Girls Birgit Berg, Birgit Berg Consultant Child & Adolescent Psychiatrist, LondonSearch for more papers by this author Birgit Berg, Birgit Berg Consultant Child & Adolescent Psychiatrist, LondonSearch for more papers by this author First published: 07 October 2004 https://doi.org/10.1111/j.1475-3588.2004.00104_12.xRead the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Volume9, Issue4November 2004Pages 199-199 RelatedInformation

A survey was carried out of psychiatrists working in the North West Thames Region to assess their knowledge and clinical practice regarding the children of mothers with eating disorders. One hundred and ten psychiatrists were contacted and they returned 66 completed questionnaires. Most psychiatrists were aware that these children might be at increased risk for difficulties. They had little knowledge concerning the children whose mothers they treated for eating disorders, and the effect of the maternal disorder on children's eating, growth and psychological adjustment. The majority did not assess or treat the family or children. A previous Placement in child and adolescent psychiatry was associated with greater awareness of possible problems for children of eating-disordered mothers (p=0.008). Adult psychiatrists are well placed to identify the psychiatric needs of the children of their patients, so continued attention should be given to their training and clinical practice.