Modulation of sulfur metabolism enables efficient glucosinolate engineering

<p>Abstract</p> <p>Background</p> <p>Metabolic engineering in heterologous organisms is an attractive approach to achieve efficient production of valuable natural products. Glucosinolates represent a good example of such compounds as they are thought to be the cancer-preventive agents in cruciferous plants. We have recently demonstrated that it is feasible to engineer benzylglucosinolate (BGLS) in the non-cruciferous plant <it>Nicotiana benthamiana </it>by transient expression of five genes from <it>Arabidopsis thaliana</it>. In the same study, we showed that co-expression of a sixth <it>Arabidopsis </it>gene, <it>γ-glutamyl peptidase 1 </it>(<it>GGP1</it>), resolved a metabolic bottleneck, thereby increasing BGLS accumulation. However, the accumulation did not reach the expected levels, leaving room for further optimization.</p> <p>Results</p> <p>To optimize heterologous glucosinolate production, we have in this study performed a comparative metabolite analysis of BGLS-producing <it>N. benthamiana </it>leaves in the presence or absence of <it>GGP1</it>. The analysis revealed that the increased BGLS levels in the presence of <it>GGP1 </it>were accompanied by a high accumulation of the last intermediate, desulfoBGLS, and a derivative thereof. This evidenced a bottleneck in the last step of the pathway, the transfer of sulfate from 3'-phosphoadenosine-5'-phosphosulfate (PAPS) to desulfoBGLS by the sulfotransferase AtSOT16. While substitution of AtSOT16 with alternative sulfotransferases did not alleviate the bottleneck, experiments with the three genes involved in the formation and recycling of PAPS showed that co-expression of <it>adenosine 5'-phosphosulfate kinase 2 </it>(<it>APK2</it>) alone reduced the accumulation of desulfoBGLS and its derivative by more than 98% and increased BGLS accumulation 16-fold.</p> <p>Conclusion</p> <p>Adjusting sulfur metabolism by directing sulfur from primary to secondary metabolism leads to a remarkable improvement in BGLS accumulation and thereby represents an important step towards a clean and efficient production of glucosinolates in heterologous hosts. Our study emphasizes the importance of considering co-substrates and their biological nature in metabolic engineering projects.</p

Danish Foundation Models

Large language models, sometimes referred to as foundation models, have transformed multiple fields of research. However, smaller languages risk falling behind due to high training costs and small incentives for large companies to train these models. To combat this, the Danish Foundation Models project seeks to provide and maintain open, well-documented, and high-quality foundation models for the Danish language. This is achieved through broad cooperation with public and private institutions, to ensure high data quality and applicability of the trained models. We present the motivation of the project, the current status, and future perspectives.Comment: 4 pages, 2 table

Mesenchymal stromal cell treatment of donor kidneys during ex-vivo normothermic machine perfusion:A porcine renal autotransplantation study

Normothermic machine perfusion (NMP) of injured kidneys offers the opportunity for interventions to metabolically active organs prior to transplantation. Mesenchymal stromal cells (MSCs) can exert regenerative and anti-inflammatory effects in ischaemia-reperfusion injury. The aims of this study were to evaluate the safety and feasibility of MSC treatment of kidneys during NMP using a porcine auto-transplantation model, and examine potential MSC treatment-associated kidney improvements up to 14 days post transplantation. After 75 minutes of kidney warm ischaemia, four experimental groups of n=7 underwent 14 hours of oxygenated hypothermic machine perfusion. In three groups this was followed by 240 minutes of NMP with infusion of vehicle, ten million porcine or ten million human adipose derived MSCs. All kidneys were auto-transplanted after contralateral nephrectomy. MSC treatment did not affect perfusion haemodynamics during NMP or cause adverse effects at reperfusion, with 100% animal survival. MSCs did not affect plasma creatinine, glomerular filtration rate, neutrophil gelatinase-associated lipocalin concentrations or kidney damage assessed by histology during the 14 days, and MSCs retention was demonstrated in renal cortex. Infusing MSCs during ex vivo NMP of porcine kidneys was safe and feasible. Within the short post- transplant

Risk sharing arrangements for pharmaceuticals: potential considerations and recommendations for European payers

<p>Abstract</p> <p>Background</p> <p>There has been an increase in 'risk sharing' schemes for pharmaceuticals between healthcare institutions and pharmaceutical companies in Europe in recent years as an additional approach to provide continued comprehensive and equitable healthcare. There is though confusion surrounding the terminology as well as concerns with existing schemes.</p> <p>Methods</p> <p>Aliterature review was undertaken to identify existing schemes supplemented with additional internal documents or web-based references known to the authors. This was combined with the extensive knowledge of health authority personnel from 14 different countries and locations involved with these schemes.</p> <p>Results and discussion</p> <p>A large number of 'risk sharing' schemes with pharmaceuticals are in existence incorporating both financial-based models and performance-based/outcomes-based models. In view of this, a new logical definition is proposed. This is "<it>risk sharing' schemes should be considered as agreements concluded by payers and pharmaceutical companies to diminish the impact on payers' budgets for new and existing schemes brought about by uncertainty and/or the need to work within finite budgets</it>". There are a number of concerns with existing schemes. These include potentially high administration costs, lack of transparency, conflicts of interest, and whether health authorities will end up funding an appreciable proportion of a new drug's development costs. In addition, there is a paucity of published evaluations of existing schemes with pharmaceuticals.</p> <p>Conclusion</p> <p>We believe there are only a limited number of situations where 'risk sharing' schemes should be considered as well as factors that should be considered by payers in advance of implementation. This includes their objective, appropriateness, the availability of competent staff to fully evaluate proposed schemes as well as access to IT support. This also includes whether systematic evaluations have been built into proposed schemes.</p

Netnography of Female Use of the Synthetic Growth Hormone CJC-1295: Pulses and Potions

Background. Communal online folk pharmacology fuels the drive for short cuts in attaining muscle enhancement, fat loss, and youthful skin. Objectives. The study used “netnography” to explore female use of CJC-1295, a synthetic growth hormone analogue from the perspectives contained in Internet forum activity. Methods. A systematic Internet search was conducted using variation of the term “CJC-1295”; and combined with “forum.” Ninety-six hits related to bodybuilding websites where CJC-1295 was mentioned. Following application of exclusion criteria to confine to female use and evidence of forum activity, 9 sites remained. These were searched internally for reference to CJC-1295. Twenty-three discussion threads relating to female use of CJC-1295 formed the end data set, and analyzed using the Empirical Phenomenological Psychological method. Results. Forum users appeared well versed and experienced in the poly use of performance and image drug supplementation. Choice to use CJC-1295 centered on weight loss, muscle enhancement, youthful skin, improved sleep, and injury healing. Concerns were described relating to female consequences of use given gender variations in growth hormone pulses affecting estimation of dosage, cycling, and long-term consequences. Conclusions. Public health interventions should consider female self-medicating use of synthetic growth hormone within a repertoire of product supplementation, and related adverse health consequences