En idéträdgård

Som student har jag blivit inbjuden att skapa en idéträdgård på Malmö garden show den 1-3/6 2012. Med mitt intresse i natur och hälsa blev det självklara valet en trädgård med fokus på just detta. En kontemplativ plats som ska inspirera människor till att med små medel skapa förutsättningar för en sundare livsstil och högre livskvalitet. Kombinationen meditation och trädgård driver tankarna genast till Japan och deras uråldriga trädgårdskultur som är huvudinspirationen till den gröna delen i gestaltningen. Den andra inspirationen kommer ur den arabiska orienten med svala platser under skuggande tyger och bland mjuka kuddar. Detta presenteras i en paviljong men med en nordisk tolkning. Anledningen till den till synes märkliga kombinationen grundar sig i alla kulturkrockar man dagligen möter i Malmö och resten av världen. Med detta sagt inte nödvändigtvis negativa krockar, utan möjligheten att skapa spännande möten på traditionella platser. I denna lugna miljö får besökaren uppleva vackra gröna växter som sänker pulsen och inger en ro i kroppen som bara naturen kan. Under ett besök till Nordiska trädgårdar på Älvsjömässan i Stockholm i april utförde jag en del intervjuer med besökare och utställare för att få tips inför mitt eget projekt. Framför allt har jag fått värdefulla råd angående den tekniska konstruktionen men även små saker att tänka på som att beställa tillräckliga mängder material vilket beskrevs som en återkommande miss bland de intervjuade. En stor del av arbetet består av tekniska beskrivningar av byggandet av trädgården, ritningar och fotografier. Den ekonomiska situationen som student innebar att allt material jag behövt har jag fått genom sponsorer och gåvor från vänner och familj. Detta har inneburit tidskrävande idogt arbete.As a student I was invited to create a conceptual garden at the event Malmö garden show that took place in June 1-3 2012. Considering my deep interest in the combination nature – health, this was my natural choice of theme. I wanted to create a contemplative place to inspire people to create the conditions for a healthier lifestyle and a higher quality of life, using small means. The combination of meditation and gardens immediately brings Japanese garden design to most minds; their ancient ways of treating nature in the garden is my main inspiration. My second inspiration comes from the Arabic orient with its cool pavilions, shade giving drapes and lots of soft cushions to rest tired legs. This I presented in a sitting area but with a Nordic interpretation or twist. The reason to the seemingly odd combination of styles is based on the daily cultural clashes in Malmö and many other places in the world. With this said not necessarily the negative kind of clashes, but the opportunity to create interesting meetings in traditional places. In this soothing environment the visitor can experience beautiful green plants that will lower their heart rate and submit a peacefulness that only nature could accomplish. During a trip to Nordiska trädgårdar (Nordic gardens) in Stockholm in April, I performed numerous interviews with visitors and exhibitors to gather information regarding my own project. Particularly good advice considering the construction and the importance of ordering enough material was a recurring mistake amongst the exhibitors. A big part of this work consists of technical descriptions building the garden, blueprints, drawings and photos. The economic situation of being a student is limiting and all the needed material I got through sponsors and friends and family. This meant time consuming, hard work

Alternative Exon Usage Selectively Determines Both Tissue Distribution and Subcellular Localization of the acyl-CoA Thioesterase 7 Gene Products.

Acyl-CoA thioesterases (ACOTs) catalyze the hydrolysis of acyl-CoAs to free fatty acids and coenzyme A. Recent studies have demonstrated that one gene named Acot7, reported to be mainly expressed in brain and testis, is transcribed in several different isoforms by alternative usage of first exons. Strongly decreased levels of ACOT7 activity and protein in both mitochondria and cytosol was reported in patients diagnosed with fatty acid oxidation defects, linking ACOT7 function to regulation of fatty acid oxidation in other tissues. In this study, we have identified five possible first exons in mouse Acot7 (Acot7a-e) and show that all five first exons are transcribed in a tissue specific manner. Taken together, these data show that the Acot7 gene is expressed as multiple isoforms in a tissue specific manner, and that expression in tissues other than brain and testis is likely to play important roles in fatty acid metabolism

Succinate receptor GPR91, a Gαi coupled receptor that increases intracellular calcium concentrations through PLCβ

AbstractSuccinate has been reported as the endogenous ligand for GPR91. In this study, succinate was confirmed to activate GPR91 resulting in both 3′–5′-cyclic adenosine monophosphate (cAMP) inhibition and inositol phosphate formation in a pertussis toxin (PTX)-sensitive manner. GPR91 agonist-mediated effects detected using dynamic mass redistribution (DMR) were inhibited with PTX, edelfosine and U73122 demonstrating the importance of not only the Gαi pathway but also PLCβ. These results show that GPR91 when expressed in HEK293s cells couples exclusively through the Gαi pathway and acts through Gαi not only to inhibit cAMP production but also to increase intracellular Ca2+ in an inositol phosphate dependent mechanism via PLCβ activation

Correction of a urea cycle defect after ex vivo gene editing of human hepatocytes

Ornithine transcarbamylase deficiency (OTCD) is a monogenic disease of ammonia metabolism in hepatocytes. Severe disease is frequently treated by orthotopic liver transplantation. An attractive approach is the correction of a patient's own cells to regenerate the liver with gene-repaired hepatocytes. This study investigates the efficacy and safety of ex vivo correction of primary human hepatocytes. Hepatocytes isolated from an OTCD patient were genetically corrected ex vivo, through the deletion of a mutant intronic splicing site achieving editing efficiencies >60% and the restoration of the urea cycle in vitro. The corrected hepatocytes were transplanted into the liver of FRGN mice and repopulated to high levels (>80%). Animals transplanted and liver repopulated with genetically edited patient hepatocytes displayed normal ammonia, enhanced clearance of an ammonia challenge and OTC enzyme activity, as well as lower urinary orotic acid when compared to mice repopulated with unedited patient hepatocytes. Gene expression was shown to be similar between mice transplanted with unedited or edited patient hepatocytes. Finally, a genome-wide screening by performing CIRCLE-seq and deep sequencing of >70 potential off-targets revealed no unspecific editing. Overall analysis of disease phenotype, gene expression, and possible off-target editing indicated that the gene editing of a severe genetic liver disease was safe and effective. Keywords: CRISPR; FRGN; ex vivo; genome editing; hepatocyte transplantation; liver-humanized mouse; primary hepatocytes; urea cycle disorder

Alternative Exon Usage Selectively Determines Both Tissue Distribution and Subcellular Localization of the acyl-CoA Thioesterase 7 Gene Products.

Acyl-CoA thioesterases (ACOTs) catalyze the hydrolysis of acyl-CoAs to free fatty acids and coenzyme A. Recent studies have demonstrated that one gene named Acot7, reported to be mainly expressed in brain and testis, is transcribed in several different isoforms by alternative usage of first exons. Strongly decreased levels of ACOT7 activity and protein in both mitochondria and cytosol was reported in patients diagnosed with fatty acid oxidation defects, linking ACOT7 function to regulation of fatty acid oxidation in other tissues. In this study, we have identified five possible first exons in mouse Acot7 (Acot7a-e) and show that all five first exons are transcribed in a tissue specific manner. Taken together, these data show that the Acot7 gene is expressed as multiple isoforms in a tissue specific manner, and that expression in tissues other than brain and testis is likely to play important roles in fatty acid metabolism

G protein-coupled receptor GPR35 suppresses lipid accumulation in hepatocytes

Although prevalent, nonalcoholic fatty liver disease is not currently treated effectively with medicines. Initially, using wild-type and genome-edited clones of the human hepatocyte cell line HepG2, we show that activation of the orphan G protein-coupled receptor GPR35 is both able and sufficient to block liver X-receptor-mediated lipid accumulation. Studies on hepatocytes isolated from both wild-type and GPR35 knock-out mice were consistent with a similar effect of GPR35 agonists in these cells, but because of marked differences in the pharmacology of GPR35 agonists and antagonists at the mouse and human orthologues, as well as elevated basal lipid levels in hepatocytes from the GPR35 knock-out mice, no definitive conclusion could be reached. To overcome this, we generated and characterized a transgenic knock-in mouse line in which the corresponding human GPR35 splice variant replaced the mouse orthologue. In hepatocytes from these humanized GPR35 mice, activation of this receptor was shown conclusively to prevent, and also reverse, lipid accumulation induced by liver X-receptor stimulation. These studies highlight the potential to target GPR35 in the context of fatty liver diseases

Pharmacological Treatment in Forensic Psychiatry-A Systematic Review

Background: Pharmacological treatment is of great importance in forensic psychiatry, and the vast majority of patients are treated with antipsychotic agents. There are several systematic differences between general and forensic psychiatric patients, e.g. severe violent behavior, the amount of comorbidity, such as personality disorders and/or substance abuse. Based on that, it is reasonable to suspect that effects of pharmacological treatments also may differ. The objective of this systematic review was to investigate the effects of pharmacological interventions for patients within forensic psychiatry. Methods: The systematic review protocol was pre-registered in PROSPERO (CRD42017075308). Six databases were used for literature search on January 11, 2018. Controlled trials from forensic psychiatric care reporting on the effects of antipsychotic agents, mood stabilizers, benzodiazepines, antidepressants, as well as pharmacological agents used for the treatment of addiction or ADHD, were included. Two authors independently reviewed the studies, evaluated risk of bias and assessed certainty of evidence using Grading of Recommendations Assessment, Development and Evaluation (GRADE). Results: The literature search resulted in 1783 records (titles and abstracts) out of which 10 studies were included. Most of the studies included were retrospective and non-randomized. Five of them focused on treatment with clozapine and the remaining five on other antipsychotics or mood stabilizers. Five studies with a high risk of bias indicated positive effects of clozapine on time from treatment start to discharge, crime-free time, time from discharge to readmission, improved clinical functioning, and reduction in aggressive behavior. Psychotic symptoms after treatment were more pronounced in the clozapine group. Mainly due to the high risk of bias the reliability of the evidence for all outcomes was assessed as very low. Conclusion: This systematic review highlights the shortage of knowledge on the effectiveness of pharmacological treatment within forensic psychiatry. Due to very few studies being available in this setting, as well as limitations in their execution and reporting, it is challenging to overview the outcomes of pharmacological interventions in this context. The frequent use of antipsychotics, sometimes in combination with other pharmacological agents, in this complex and heterogeneous patient group, calls for high-quality studies performed in this specific setting

ULK1 knockout cell line downregulates autophagy, upregulates recombinant transcript and improves protein secretion

To meet the ever-growing demand for effective, safe, and affordable protein therapeutics, decades of bioprocessing innovations and cell engineering modifications have vastly improved the production of recombinant proteins. Recently, new genomic technologies allow more targeted approaches in cell line development where most effort have been aimed at reducing toxic byproducts or regulate specific traits, such as apoptosis or glycosylation. By targeting a general metabolic or processing pathway a broader protein expression increase could be achieved. In order to identify a more general cell engineered platform, a small molecule screen for enhanced protein expression of three model proteins was performed. From this, ULK1, the key initiator of autophagy, emerged as an important component for improved recombinant expression. Autophagy is a large process within the cell that restores cell homeostasis during cell stress, and knockout of ULK1 improved protein expression 3-fold in a stable Cripto-Fc producing cell. Transcriptomic analysis showed a downregulation of autophagy and upregulation of transcriptional processes when ULK1 was removed. Processes within the host cell that are of a general cell maintenance character have great potential to be engineered into a universal manufacturing platform, here shown through the prevention of autophagy.QC 20210604</p