Marine DNA polymerases as tools for next generation molecular diagnostics solutions

Globally, there is a high demand to monitor and diagnose critical infectious diseases. Current molecular diagnostic tests require long “sample in - answer out” time and expensive equipment. Isothermal Nucleic Acid Amplification Technology (INAAT) is an enabling technology possibly offering rapid, sensitive and specific molecular diagnosis of infectious diseases. However, the implementation of INAAT into Point of Care diagnostic platforms are halted due to a very limited selection of suitable enzymes, i.e. polymerases, performing at ambient temperatures. The overall goal with the project is to bring a panel of new and improved polymerase enzymes to market for use in next generation INAAT molecular diagnostic platforms and other potential application areas, e.g. next-generation sequencing and single-cell genomics. We have identified and characterized a large selection of polymerase enzymes based on marine bioprospecting of the arctic and subarctic region. These act as starting point for the development of 2nd generation isothermal amplification polymerases. Through use of our highly versatile molecular evolution library, combined with rational design studies, tailor-made enzymes for various application areas will be developed. This project emphasizes on early market feedback to confirm relevance and applicability and to ensure that enzymes are developed according to customer needs

Sensory-Based Interventions in Inpatient Mental Health

This systematic review aimed to answer the following question: What are effective sensory-based interventions that increase self-regulation skills and decrease maladaptive behaviors for adults in inpatient mental health settings? Researchers collaborated with local occupational therapy practitioner, Katrina LaRossa, from a Multicare Behavioral Health Hospital in Western Washington. Sixteen articles met the inclusion criteria and were rigorously reviewed and categorized by the researchers. Evidence from the literature indicates that sensory-based interventions can be an inexpensive and effective method to reduce agitation and maladaptive behaviors. The knowledge translation project involved development of a guide that provided implementation strategies for the sensory-based interventions identified in the research process. The implementation guide was organized in a binder, and included the implementation methods, outcomes, credibility, cost breakdown, and special considerations for each sensory-based intervention, as well as the article itself. To monitor the impact of our knowledge translation project, a survey was conducted to measure the clinician’s perception and satisfaction with the implementation guide. Researchers recommend that future research focuses on sensory-based interventions for adults in inpatient mental health and that studies take into account restrictions that are typical for inpatient mental health

Døgnvariasjoner for innleggelser ved akuttpsykiatriske enheter

Akuttpsykiatriske enheter gir øyeblikkelig hjelp til pasienter med særlig vanskelige og kompliserte tilstander med behov utover det som kan ivaretas i kommunehelsetjenesten eller ved distriktspsykiatriske senter (DPS). Flere studier har de siste årene sett på antallet innleggelser og hyppigheten av tvangsbruk i akuttpsykiatriske enheter i Norge (Bjerke et al., 2019; Tøgersen et al., 2015; Wynn, 2018). Bjerke, Gjelstad og Ruud viser at antallet tvangsinnleggelser holdt seg stabilt i perioden 2010 til 2017 (Bjerke et al., 2019). Videre er det funnet at den gjennomsnittlige liggetiden i psykisk helsevern er redusert med 30 % i perioden fra 2009 til 2018 (Statistisk sentralbyrå, 2019). Noen studier har også sett på tidspunkt for innleggelser i akuttpsykiatrien, det vil si når på døgnet pasienter blir lagt inn. Dette har imidlertid ikke blitt undersøkt de siste 10 årene og etter at samhandlingsreformen trådte i kraft (St.meld. nr. 47, 2008). Innleggelser ved akuttpsykiatriske enheter viser noe ulike mønstre. En norsk multisenterstudie fra 2006 der 19 akuttpsykiatriske enheter deltok, viste at gjennomsnittlig 48 % av innleggelsene i akuttpsykiatrien fant sted på dagtid, mens 42 % på kveldstid og 10 % på natt (Gråwe, Hatling & Ruud, 2006). En annen norsk studie fant at 24 % av pasientene ble innlagt mellom klokken (kl.) 16:00 og 22:00, mens 19 % ble lagt inn mellom kl. 22:00 og kl. 08:00, og øvrige innleggelser fant sted på dagtid (Deraas et al., 2006). Videre viste en undersøkelse av 1323 psykiatriske akuttinnleggelser på Lovisenberg Sykehus at 52 % av innleggelsene fant sted på dagtid, altså mellom klokken 08:00 og 16:00 (Berg, 2007). Det ble funnet at 39 % ble innlagt mellom klokken 16:00 og 01:00 og 9 % i tidsrommet 01:00–08:00. Forskjellene mellom tidspunktene var imidlertid ikke signifikante. Hensikten med den nåværende studien er å undersøke hvordan innleggelser i akuttpsykiatriske enheter fordeler seg over døgnet. Vi forventet å finne at flertallet av innleggelsene skjedde på dagtid

A mitochondria-targeted mass spectrometry probe to detect glyoxals: implications for diabetes

The glycation of protein and nucleic acids that occurs as a consequence of hyperglycaemia disrupts cell function and contributes to many pathologies, including those associated with diabetes and aging. Intracellular glycation occurs following the generation of the reactive 1,2-dicarbonyls methylglyoxal and glyoxal and disruption to mitochondrial function is associated with hyperglycemia. However, the contribution of these reactive dicarbonyls to mitochondrial damage in pathology is unclear due to uncertainties about their levels within mitochondria in cells and in vivo. To address this we have developed a mitochondria-targeted reagent (MitoG) designed to assess the levels of mitochondrial dicarbonyls within cells. MitoG comprises a lipophilic triphenylphosphonium cationic function, which directs the molecules to mitochondria within cells and an o-phenylenediamine moiety that reacts with dicarbonyls to give distinctive and stable products. The extent of accumulation of these diagnostic heterocyclic products can be readily and sensitively quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS), enabling changes to be determined. Using the MitoG-based analysis we assessed the formation of methylglyoxal and glyoxal in response to hyperglycaemia in cells in culture and in the Akita mouse model of diabetes in vivo. These findings indicated that the levels of methylglyoxal and glyoxal within mitochondria increase during hyperglycaemia in both cells and in vivo, suggesting that they can contribute to the pathological mitochondrial dysfunction that occurs in diabetes and aging

Identification of the decumbenone biosynthetic gene cluster in and the importance for production of calbistrin

Background: Filamentous fungi are important producers of secondary metabolites, low molecular weight molecules that often have bioactive properties. Calbistrin A is a secondary metabolite with an interesting structure that was recently found to have bioactivity against leukemia cells. It consists of two polyketides linked by an ester bond: a bicyclic decalin containing polyketide with structural similarities to lovastatin, and a linear 12 carbon dioic acid structure. Calbistrin A is known to be produced by several uniseriate black Aspergilli, Aspergillus versicolor-related species, and Penicillia. Penicillium decumbens produces calbistrin A and B as well as several putative intermediates of the calbistrin pathway, such as decumbenone A-B and versiol. Results: A comparative genomics study focused on the polyketide synthase (PKS) sets found in three full genome sequence calbistrin producing fungal species, P. decumbens, A. aculeatus and A. versicolor, resulted in the identification of a novel, putative 13-membered calbistrin producing gene cluster (calA to calM). Implementation of the CRISPR/Cas9 technology in P. decumbens allowed the targeted deletion of genes encoding a polyketide synthase (calA), a major facilitator pump (calB) and a binuclear zinc cluster transcription factor (calC). Detailed metabolic profiling, using UHPLC-MS, of the ∆calA (PKS) and ∆calC (TF) strains confirmed the suspected involvement in calbistrin productions as neither strains produced calbistrin nor any of the putative intermediates in the pathway. Similarly analysis of the excreted metabolites in the ∆calB (MFC-pump) strain showed that the encoded pump was required for efficient export of calbistrin A and B. Conclusion: Here we report the discovery of a gene cluster (calA-M) involved in the biosynthesis of the polyketide calbistrin in P. decumbens. Targeted gene deletions proved the involvement of CalA (polyketide synthase) in the biosynthesis of calbistrin, CalB (major facilitator pump) for the export of calbistrin A and B and CalC for the transcriptional regulation of the cal-cluster. This study lays the foundation for further characterization of the calbistrin biosynthetic pathway in multiple species and the development of an efficient calbistrin producing cell factory