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Here, we describe a detailed step-by-step protocol for the expression, purification, quantification, and activity determination of key enzymes for molecular detection of pathogens. Based on previous reports, we optimized the protocol for LbCas12a, Taq DNA polymerase, M-MLV reverse transcriptase, and TEV protease to make it compatible with minimal laboratory equipment, broadly available in low- and
During nutrient limitation, bacteria produce the alarmones (p)ppGpp as effectors of a stress signaling network termed the stringent response. RsgA, RbgA, Era, and HflX are four ribosome-associated GTPases (RA-GTPases) that bind to (p)ppGpp in Staphylococcus aureus. These enzymes are cofactors in ribosome assembly, where they cycle between the ON (GTP-bound) and OFF (GDP-bound) ribosome-associated
Low- and middle-income countries (LMICs) are significantly affected by SARS-CoV-2, partially due to their limited capacity for local production and implementation of molecular testing. Here, we provide detailed methods and validation of a molecular toolkit that can be readily produced and deployed using laboratory equipment available in LMICs. Our results show that lab-scale production of enzymes
Here, we describe a detailed step-by-step protocol to detect SARS-CoV-2 RNA using RT-PCR-mediated amplification and CRISPR/Cas-based visualization. The optimized assay uses basic molecular biology equipment such as conventional thermocyclers and transilluminators for qualitative detection. Alternatively, a fluorescence plate reader can be used for quantitative measurements. The protocol detects tw
Initiation factor IF3 is an essential protein that enhances the fidelity and speed of bacterial mRNA translation initiation. Here, we describe the dynamic interplay between IF3 domains and their alternative binding sites using pre-steady state kinetics combined with molecular modelling of available structures of initiation complexes. Our results show that IF3 accommodates its domains at velocities
Rapid Diagnostic Tests (RDTs) for malaria are restricted to a few biomarkers and antibody-mediated detection. However, the expression of commonly used biomarkers varies geographically and the sensibility of immunodetection can be affected by batch-to-batch differences or limited thermal stability. In this study we aimed to overcome these limitations by identifying a potential biomarker and by deve
During host colonization, bacteria use the alarmones (p)ppGpp to reshape their proteome by acting pleiotropically on DNA, RNA, and protein synthesis. Here, we elucidate how the initiating ribosome senses the cellular pool of guanosine nucleotides and regulates the progression towards protein synthesis. Our results show that the affinity of guanosine triphosphate (GTP) and the inhibitory concentrat
It is common in elite sport to monitor athletes' training load, injuries and illnesses, but mental distress is rarely included. An improved understanding of the epidemiology of mental distress among elite Para athletes and how their coaches perceive such monitoring would allow us to better develop and implement preventive measures. The purpose of this study was therefore to (1) prospectively descr
This study explored all-cause mortality of bacteremia diagnosed during a 60-day non-physician healthcare worker strike in 2008. A significant change, with 5.0% (95% confidence interval [CI] 1.2-8.7%, P < .01) absolute risk increase, was seen in 90-day mortality during the strike (n = 598) compared with the rest of the study period 2000-2015 (n = 75 647).
Funding is central to the viability of a class action regime. Wherever class actions have been adopted, legislators have used an array of design points to shape regimes that facilitate the requisite economic enablers and conditions. Yet one facet that has received insufficient attention in research and reforms is the potential of a public funder to advance collective claims and promote public inte
Introduces a special issue on the development of the class action regime, in light of the reforms reflected in the Supreme Court decisions in Mastercard Inc v Merricks and Google LLC v Lloyd, and a conference hosted by the Bonavero Institute of Human Rights at the University of Oxford entitled "Access to Justice and Class Actions in England and Wales".
Idén till den här antologin föddes för flera år sedan, i samband med densenaste processen kring legitimation för kuratorer inom hälso- och sjuk-vården. Under våren 2018, efter att fackförbundet SSR vid ett flertal tillfäl-len sedan 1967 initierat och drivit frågan, fattade Utbildningsdepartementet,i enlighet med Socialstyrelsens bedömning (2014), slutligen beslutet attinföra legitimation. I juli 2
Class actions in Sweden are not flourishing. This paper examines why the oldest class action regime in Scandinavia has not developed into anything more than a peripheral addition to the Swedish legal landscape, drawing on relevant procedural, political, and sociological insights. It is divided into two sections that (1) analyse the reform process in the 1990s and latest developments, including mai
An album released by Carrier Records. It includes performances and compositions and videographic works by Davíð Brynjar Franzson, Luong Hue Trinh, Kent Olofsson, Halla Steinunn Stefánsdóttir and Mirjam Tally. Information by Stefánsdóttir, taken from the album website:strengur is a set of five groups of three pieces, each group created in deep collaboration with a long-term creative partner. For th
This article approaches listening practices and the role of technological mediation within ecological sound art, building on findings through the artistic research practices of the two authors. Through documentation of the authors’ ecological sound art practices of aeolian guitar performance, curation, composition, performance on found objects and field recording, we argue that phenomenological va
A tree is so much more than just a tree! Trees provide habitat to support biodiversity, remove carbon dioxide and particulate matter from our air, cool our cities during summers, and inspire enjoyment by all. A tree planted deliberately to provide any of these services is considered a nature-based solution. In this episode, we present the concept of nature-based solutions as an approach to reduce
Aminoglycoside antibiotics are widely used to treat infectious diseases. Among them, streptomycin and kanamycin (and derivatives) are of importance to battle multidrug-resistant (MDR) Mycobacterium tuberculosis. Both drugs bind the small ribosomal subunit (30S) and inhibit protein synthesis. Genetic, structural, and biochemical studies indicate that local and long-range conformational rearrangemen
While Weighted Lasso sparse regression has appealing statistical guarantees that would entail a major real-world impact in finance, genomics, and brain imaging applications, it is typically scarcely adopted due to its complex high-dimensional space composed by thousands of hyperparameters. On the other hand, the latest progress with high-dimensional hyperparameter optimization (HD-HPO) methods for
Gruv- och ballastindustrin utgör en mycket viktig sektor i den svenska ekonomin. Där gruvindustrin operar på den globala arenan medan sten branschen operar på regional/lokal basis. Efterfrågan av metaller- och byggnadsmaterial har ökat under det senaste decenniet och mängden fasta mineraler och stenar som tas ut ur jordskorpan är nästan 80 miljarder ton per år, med ballast som den största delen.DeThe mining and quarrying industries represent a highly important sector of the Swedish economy. The mining industry operates globally, whereas the quarrying industry operates on a regional/local basis. Demands for metal and construction aggregates have increased over the last decade, and almost 80 billion tons of solid minerals and rocks are extracted from the earth’s crust each year. Construction
