Gutierrezguldager0860

Biological samples usually require cumbersome preparation steps before SEM imaging. Here we propose a simple, fast and inexpensive method to prepare and visualize biological cell culture samples in a few easy steps. We have tested this method with success on several adherent breast cancer and non-adherent leukemia cell lines. This method gives results comparable to other well-established techniques, and it can be convenient in day-to-day biological sample preparation for SEM imaging.•An easy and rapid method to visualize biological specimens under SEM.•Cells are grown on carbon tapes and gold coated.•Air drying without compromising the image quality.Mitochondria generate aerobic cellular energy (i.e., ATP) through the reduction of oxygen to water via oxidative phosphorylation. The efficiency of this pathway can be measured by the phosphate/oxygen (ATP/O) ratio, which is the amount of ATP produced per oxygen atom reduced. This ratio thus provides a measure of the efficiency of mitochondrial respiration that can be readily compared between species. The magnesium green (MgGr) fluorometric method permits easy measurement of ATP/O ratios from isolated mitochondria but the standard analysis approach employs an endpoint method to calculate ATP/O ratios. Here, we present a modified method of ATP/O calculation that permits dynamic observation of fluorescent measurements of the consumption of O2 (JO2) and the production of ATP (JATP). Specifically, by substituting the slope of a straight line within a given period of time (seconds to minutes) with the slope of a tangent to each time point (per second), it is possible to evaluate JO2, JATP and the ATP/O ratio in a dynamic manner.•Provides second-by-second visualization of ATP/O ratios throughout experiments vs. a single measurement.•Dynamic visualization allows for easy identification of outlying data and more accurate calculation of mean ATP/O ratios.Many agricultural research and development programs aiming at enhancing tradeoffs related to different adoption, management and policy decisions face a methodological problem in which multi-criteria ranking is used to reach acceptable compromises between different objectives (e.g. those of farms, research managers, donors or policy makers). A typical situation is where many farm management options will result in different conflicting economic, social and environmental impacts. Ranking these options and the choice of those to promote is challenging. The literature provides a set of methodological solutions that need background data organization and simulation through coding using different computing software. Here, we provide a generic solution and friendly interface, made on Shiny (an R-package) based on the Technique for Order of Preference by Similarity to Ideal Solution (TOPSIS). We apply this method for ranking different crop technological products of grain legumes and dry cereals based on their respective impacts on poverty, child malnutrition and economic benefits in more than 40 countries in eight different geographic zones across South Asia and Sub-Saharan Africa. • The developed algorithms and interface can help rank different options based on the weights (preferences) of their respective outcome indicators. • The interface allows for changing the weights (preferences) and automatically generates new ranking tables and graphs accordingly, which can serve for scenario simulations, which saves time compared to manually performing these calculations.This paper evaluated the efficiency of beach hatcheries as a conservation tool for threatened sea turtle clutches. During six nesting seasons (2013 to 2018), several thousand high-risk clutches from loggerhead turtles (Caretta caretta) were relocated to a hatchery constructed on the same beach, within the Sea Turtle Natural Reserve (STNR, Boa Vista Island, Cabo Verde). Some parameters like hatching success; incubation period, hatchlings' morphology and their behavioral response were compared to in-situ clutches.•Our findings confirmed that the in-situ nests within the STNR had extremely high egg mortality that was usually over 70 %. Mean hatching success of clutches relocated to hatcheries was significantly higher than in-situ clutches with mean values between 70 to 85 % (p 0.05) between relocated and in-situ clutches.•This study provided a detailed method and recommendations for sea turtle clutches relocation to the hatchery, that can be beneficial for endangered sea turtle population specially where hatching success is very low.Production of a protein of interest in bacteria and its purification from bacterial lysates are valuable tools for the purification of larger amounts of recombinant proteins. The low cost of culturing, and the rapid cell growth of bacteria make this host a good choice for protein production, but the folding and function of the purified protein might be altered due to the production of a eukaryotic protein in a prokaryotic host. Here, we provide a purification method for the purification of gamma (γ)-tubulin (TUBG) from soluble fractions of Escherichia (E.) coli lysates using affinity tags.•This protocol describes a method that purifies soluble GST-TUBG1 from bacteria.•Of the three tested induction conditions, the highest yield of recombinant GST-TUBG1 was obtained after the induction of E. coli with isopropyl-D-1-thiogalactopyranoside (IPTG) for 1 h at 37 °C followed by overnight incubation at room temperature.•In comparison with other methodologies (Hoog et al., 2011), the technique described here retrieves larger amounts of recombinant TUBG1 from small-scale expression cultures.Microplastics (MP) are a pollutant that can be found in all marine ecosystems. selleck products Currently one of the most used forms to classify them is through their size. However, the current size categories in use cover an extremely wide range of sizes and are not based on a biological or physical basis. Thus, here we propose to harmonize the MP size categories with the ones already in used on plankton research for more than 120 years. This will allow the implementation of more refined MP size classes that are connected to a biological reality and will also enable the comparison of a myriad of literature on plankton research with the new work on MP.In vitro stem cell models are used as alternatives to animal models and are important tools for cytotoxicity studies. Researchers can determine the effects of test substances on human cells by evaluating cell viability and differentiation. Here, we describe an in vitro model to quantify adipogenesis based on the Nile red staining of specific lipid droplets and the emission of basic lipids from human adipose tissue-derived mesenchymal stromal cells (AD-MSCs) in the presence of test substances. This assay allows for the prediction of toxicity based on the inhibition of adipogenesis in vitro in a 96-well format. The differentiation of a progenitor cell into a specialized cell, the adipocyte, is easy to monitor and quantify, making this a simple assay. The fluorescence staining of nuclei and lipid droplets is measured after 14 days of cell differentiation to determine cell number and assess cell differentiation using high-content imaging analysis, thus allowing for the identification of chemicals that impact differentiation. We also describe a protocol to assess adipocyte differentiation by fluorescence intensity using a multiplate reader.•Researchers can utilize the protocol described here for many purposes to evaluate in vitro adipogenesis.•With this method, it is possible to reduce the use of animals.Fresh horticultural products are exposed to different environmental conditions from farm to fork. Barrier properties of packaging and physiological behaviour of produce, namely respiration and transpiration can affect headspace conditions surrounding produce and consequently remaining shelf life. Packaging material also plays a role in heat and mass transfer, such as thermal conduction and permeation of O2, CO2 and water vapour. All of these behaviours are integrated together in the form of ordinary differential equations and solved using numerical methods in MATLAB. • The simulation program is useful for designing the size and number of perforations to achieve equilibrium modified atmosphere alone or in combination with packaging material having a higher water transmission rate or active moisture absorber. • The simulation program is also useful for predicting the shelf life of fresh produce under the actual supply chain conditions. • The simulation program provides a flexible system to input predefined supply chain conditions and the properties of fresh produce and packaging material, thus, minimizing the costly and time consuming experimental procedures for selecting the optimum packaging material for fresh produce.In this paper we provide a detailed description of the methodological steps involved in conducting a Service Design study in combination with Discrete Choice Experiments (DCEs). It complements the conceptual and epistemological argument developed for this methodological combination in Osborne et al. (2021, World Development, in review WD-19535). Service Design for the co-creative development of policy interventions in complex adaptive systems involves an iterative process of moving between the six methodological stages of (1) problem co-definition, (2) actor-centred mapping, (3) experience-based problem diagnosis, (4) rapid prototyping, (5) design and testing and (6) upscaling. We suggest using DCEs as a quantitative method that is contextually adaptable and comparatively fast and cheap to implement, as part of stage (6) design and testing. Whilst both methods can operate independently with their own strengths and limitations, we find their combination to add value to the processes and outcomes of each. We illustrate the general methodological approach with a step-by-step description of its application to Weather Index Insurance in eastern Uganda. Bullet points • Service Design co-creatively develops well-targeted solutions in complex adaptive systems. • Discrete Choice Experiments quantitatively elicit actors' preferences over the design of goods or services. • Their combination can bring deeply contextualised, user-centred, operational and experimentally verified ideas for development interventions prior to their implementation.The semenogelin 1 protein is secreted in the seminal vesicles. After ejaculation it is split into small peptide fragments using internal proteases. It was shown that the fragments SEM1(45-107), SEM1(49-107), SEM1(68-107) (SEM1(86-107) form amyloid fibrils, which increase the possibility of HIV infection. The article presents a protocol for the synthesis and purification of a 15N, 13C-labeled SEM1(68-107) peptide for further structural studies by high-resolution NMR spectroscopy. The work describes cloning, expression of fusion protein GB1-SEM1(68-107) in E.coli, its purification, removal of GB1 and purification of SEM1(68-107). The purity of SEM1(68-107) samples on each purification steps was evaluated by polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE) and tricine-SDS-PAGE. The developed protocol allows to obtain SEM1(68-107) peptide for NMR studies (using 3D experiments), instead of costly solid-phase synthesis.