Modular detergents tailor the purification and structural analysis of membrane proteins including G-protein coupled receptors

Detergents enable the purification of membrane proteins and are indispensable reagents instructural biology. Even though a large variety of detergents have been developed in the lastcentury, the challenge remains to identify guidelines that allowfine-tuning of detergents forindividual applications in membrane protein research. Addressing this challenge, here weintroduce the family of oligoglycerol detergents (OGDs). Native mass spectrometry (MS)reveals that the modular OGD architecture offers the ability to control protein purificationand to preserve interactions with native membrane lipids during purification. In addition to abroad range of bacterial membrane proteins, OGDs also enable the purification and analysisof a functional G-protein coupled receptor (GPCR). Moreover, given the modular design ofthese detergents, we anticipatefine-tuning of their properties for specific applications instructural biology. Seen from a broader perspective, this represents a significant advance forthe investigation of membrane proteins and their interactions with lipids

Only small changes in soil organic carbon and charcoal concentrations found one year after experimental slash-and-burn in a temperate deciduous forest

International audienceAnthropogenic fires affected the temperate deciduous forests of Central Europe over millennia. Biomass burning releases carbon to the atmosphere and produces charcoal, which potentially contributes to the stable soil carbon pools and is an important archive of environmental history. The fate of charcoal in soils of temperate deciduous forests, i.e. the processes of charcoal incorporation and transportation, and the effects on soil organic matter are still not clear. In a long-term experimental burning site, we investigated the effects of slash-and-burn and determined soil organic carbon, charcoal carbon and nitrogen concentrations and the soil lightness of colour (L*) in the topmost soil material (0?1, 1?2.5 and 2.5?5 cm depths) before, immediately after the fire and one year after burning. The main results are that (i) only few charcoal particles from the forest floor were incorporated into the soil matrix by soil mixing animals. In 0?1 cm and during one year, the charcoal C concentrations increased only by 0.4 g kg?1 and the proportion of charcoal C to SOC concentrations increased from 2.8 to 3.4%; (ii) the SOC concentrations did not show any significant differences; (iii) soil lightness significantly decreased in the topmost soil layer and correlated with the concentrations of charcoal C (r=-0.87**) and SOC (r=?0.94**) in samples 0?5 cm. We concluded that the soil colour depends on the proportion of aromatic charcoal carbon in total organic matter and that Holocene burning could have influenced soil charcoal concentrations and soil colour

The splicing landscape is globally reprogrammed during male meiosis

Meiosis requires conserved transcriptional changes, but it is not known whether there is a corresponding set of RNA splicing switches. Here, we used RNAseq of mouse testis to identify changes associated with the progression from mitotic spermatogonia to meiotic spermatocytes. We identified ∼150 splicing switches, most of which affect conserved protein-coding exons. The expression of many key splicing regulators changed in the course of meiosis, including downregulation of polypyrimidine tract binding protein (PTBP1) and heterogeneous nuclear RNP A1, and upregulation of nPTB, Tra2β, muscleblind, CELF proteins, Sam68 and T-STAR. The sequences near the regulated exons were significantly enriched in target sites for PTB, Tra2β and STAR proteins. Reporter minigene experiments investigating representative exons in transfected cells showed that PTB binding sites were critical for splicing of a cassette exon in the Ralgps2 mRNA and a shift in alternative 5′ splice site usage in the Bptf mRNA. We speculate that nPTB might functionally replace PTBP1 during meiosis for some target exons, with changes in the expression of other splicing factors helping to establish meiotic splicing patterns. Our data suggest that there are substantial changes in the determinants and patterns of alternative splicing in the mitotic-to-meiotic transition of the germ cell cycle

Modular detergents tailor the purification and structural analysis of membrane proteins including G-protein coupled receptors

Detergents enable the purification of membrane proteins and are indispensable reagents in structural biology. Even though a large variety of detergents have been developed in the last century, the challenge remains to identify guidelines that allow fine-tuning of detergents for individual applications in membrane protein research. Addressing this challenge, here we introduce the family of oligoglycerol detergents (OGDs). Native mass spectrometry (MS) reveals that the modular OGD architecture offers the ability to control protein purification and to preserve interactions with native membrane lipids during purification. In addition to a broad range of bacterial membrane proteins, OGDs also enable the purification and analysis of a functional G-protein coupled receptor (GPCR). Moreover, given the modular design of these detergents, we anticipate fine-tuning of their properties for specific applications in structural biology. Seen from a broader perspective, this represents a significant advance for the investigation of membrane proteins and their interactions with lipids

A prospective clinical trial on the influence of a triamcinolone/demeclocycline and a calcium hydroxide based temporary cement on pain perception

<p>Abstract</p> <p>Introduction</p> <p>The aim of this clinical trial was to compare the degree of short term post-operative irritation after application of a triamcinolone/demeclocycyline based or a calcium hydroxide based provisional cement.</p> <p>Methods</p> <p>A total of 109 patients (55 female and 54 male; mean age: 51 ± 14 years) with primary or secondary dentinal caries were randomly assigned to the two treatment groups of this biomedical clinical trial (phase III). Selection criteria were good systemic health and treated teeth, which were vital and showed no symptoms of pulpitis. Up to three teeth were prepared for indirect metallic restorations, and the provisional restorations were cemented with a triamcinolone/demeclocycyline (Ledermix) or a calcium hydroxide (Provicol) based material. The intensity of post-operative pain experienced was documented according to the VAS (4, 12, 20, 24, and 82 h) and compared to VAS baseline.</p> <p>Results</p> <p>A total of 159 teeth were treated (Ledermix: 83 teeth, Provicol: 76 teeth). The minor irritation of the teeth, experienced prior to treatment, was similar in both groups; however, 4 h after treatment this value was significantly higher in the Provicol group than in the Ledermix group (p < 0.005, t-test). After 12 h, the difference was no longer significant. The number of patients taking analgesics for post-treatment pain was higher in the Provicol group (n = 11/53) than in the Ledermix group (n = 3/56).</p> <p>Conclusions</p> <p>The patients had no long term post-operative pain experience in both groups. However, within the first hours after cementation the sensation of pain was considerably higher in the Provicol group than in the Ledermix group.</p

Franchise Business Development Model: Theoretical Considerations

Franchising is not a popular business development form in Lithuania. Only 0.02% of all Lithuanian companies use franchising for business development, while in most of the developed countries franchise is recognized as a convenient business expansion form. So, what factors determine such differences in the use of franchising? Analysis of related literature showed, that mainly researchers analyse some aspects of franchising, however there is no systematized analysis which covers all the main franchise development elements. Therefore, the aim of this article is to systematize franchise business development phases, main elements and factors, influencing this process and to propose a theoretical franchise system development model. Thus, theoretical considerations about the main phases and elements of franchise system development as well as advantages and disadvantages of the system are analysed in the article. These theoretical considerations are generalized in theoretical franchise system development model, which shows, that development of franchise business consists of particular steps, starting from favourable conditions for the system to emerge; intention and ability of franchisor to form franchise based on his business; the establishment and initial development of franchise system; and its further functioning. The article does not test the model and even does not discuss the operationalization of it. Operationalization of the model as well as empirical evidence is presented by the authors in the forthcoming articles

Opening up FAIR in-situ land-use reference data: current gaps, obstacles and future challenges

It is becoming increasingly obvious that in order to address current global challenges and achieve the SDGs in the land-use sector, monitoring and evaluation using remote sensing technologies are essential. In particular, with the Copernicus program of the European Union, unprecedented free and open Earth observation data are becoming available. However, in order to improve our remotely sensed based machine learning models, training data in the form of in-situ or annotated land-use or land cover data which are based on the visual interpretation of aerial photographs or very high resolution satellite data are of utmost importance. Without sufficient training data, many land-use and land cover maps lack sufficient quality. The presentation will provide an overview of existing and open in-situ data in the field of land-use science. It will highlight what land-use data are currently available including data collected though crowdsourcing and the Geo-Wiki toolbox. In particular, it will provide insights into current gaps in land cover, land-use, livestock, forest as well as crop type information globally. It will draw on existing global data products such as those from the Copernicus global land monitoring service, and more recently generated products such as WorldCover and WorldCereal. Furthermore, tools to close those data gaps will be shown. The presentation will furthermore explore current obstacles and limitations to data sharing and debunk current arguments that are often put forth for not sharing in-situ data. These arguments include limited resources, quality issues, competition, as well as time constraints, etc. Specific attention will be given to the role of doners and funders in more clearly defining open and FAIR requirements for in-situ data. The presentation will close by making the audience aware of the LUCKINet consortium, which is trying to make more reference data openly accessible and to build a consistent global land-use change dataset as well as work done on in-situ data within the EU LAMASUS and OEMC project

Fitness, motor competence and body composition as correlates of adolescent neck/shoulder pain: an exploratory cross-sectional study

<p>Abstract</p> <p>Background</p> <p>Adolescent neck/shoulder pain (NSP) is a common and sometimes debilitating problem. Several risk factors for this condition have been investigated, but no studies have previously evaluated associations between fitness, motor competence, body composition and adolescent NSP.</p> <p>Methods</p> <p>1608 males and females of mean age 14 years answered questions on their history of NSP (4 measures), and were tested for aerobic fitness, upper and lower limb power, trunk endurance, grip strength, shoulder flexibility, motor competence and anthropometric factors. Univariate and multivariate logistic regressions were used to test for associations between NSP and physical variables.</p> <p>Results</p> <p>There were significant gender differences for most physical and pain variables. After multivariate analysis, males had lower odds of NSP if they had reduced back endurance [OR: 0.66 (95% CI: 0.46–0.97)], reduced persistent control [0.42 (0.19–0.95], and increased muscle power [0.33 (0.12–0.94)], and higher odds of NSP if they had a higher basketball throw [2.47 (1.22–5.00)] and jump performance [3.47 (1.55–7.74)]. Females had lower odds for NSP if they had a reduced jump performance [0.61(0.41–0.92)], a better basketball throw [0.60(0.40–0.90)], lower shoulder flexibility [0.54 (0.30–0.98)] and a higher aerobic capacity [0.61 (0.40–0.93)], and higher odds for NSP if they had greater abdominal endurance [1.57(1.07–2.31)] and greater bimanual dexterity [1.77(1.18–2.65)]. Females showed a U shaped relationship between NSP and back endurance [low: 2.12 (1.20–3.74); high 2.12 (1.18–3.83)].</p> <p>Conclusion</p> <p>Adolescent NSP was associated with fitness and motor competence, although the associations varied with gender, and their strength was limited.</p