vital christianity a textbook on god man cosmology faith power and spiritual science LINK 1 ENTER SITE >>> http://gg.gg/12e1hp <<< Download LINK 2 ENTER SITE >>> http://chilp.it/728ceb2 <<< Download PDF File Name:vital christianity a textbook on god man cosmology faith power and spiritual science.pdf Size: 4442 KB Type: PDF, ePub, eBook Uploaded: 10 May 2019, 21:27 Rating: 4.6/5 from 668 votes. Status: AVAILABLE Last checked: 6 Minutes ago! eBook includes PDF, ePub and Kindle version In order to read or download vital christianity a textbook on god man cosmology faith power and spiritual science ebook, you need to create a FREE account. ✔ Register a free 1 month Trial Account. ✔ Download as many books as you like (Personal use) ✔ Cancel the membership at any time if not satisfied. ✔ Join Over 80000 Happy Readers vital christianity a textbook on god man cosmology faith power and spiritual science Make a drawing. 6. Add ONE DROP of HCl (hydrochloric acid) to your beaker. 7. Record the pH. 8. Continue this until you see coacervate droplets form as a white precipitate. 9. Add a drop of this mixture to a microscope slide. Make a drawing. 10. Add a drop of methylene blue onto the slide and observe the coacervate s. Make a drawing. Do the coacervate s respond to the methylene blue stain the same way your own cheek cell would respond ? 11. Prepare a slide of one of your own cheek cells. Use methylene blue. Is this similiar to the coacervate response. Make a drawing 12. Continue adding HCl to the original mixture until there is a pH of 2. Do you see any evidence of coacervate s in the beaker. Page 2: questions. 13. Do coacervates exist Thank you, for helping us keep this platform clean. The editors will have a look at it as soon as possible. Learn about Easel TOOLS Easel Activities Pre-made digital activities. Add highlights, virtual manipulatives, and more. Browse Easel Activities Easel Assessments Quizzes with auto-grading that will be available for purchase on TpT soon. Resources are available in BOTH printable and digital formats. This curriculum bundle includes the PowerPoint presentations, notes, labs, homework assignments, task cards, daily quizzes, activities, worksheets, and unit tests for the 20 units listed below. Many scientists believe that the first cells were simply aggregates of organic molecules called coacervates. In this lab, students will first test and observe the effect of pH on coacervate formation. In the second part of the activity, students must design their own experiment to test the effect of a different variable on coacervate formation. This resource is two lab activities in one download. In the first lab activity: Students will follow a procedure to learn how coacervates are made, and will determine the ideal pH for coacervate formation. http://anvlaw.com/userfiles/bt-6500-user-manual.xml vital christianity a textbook on god man cosmology faith power and spiritual science. In the second lab activity: Students will design and carry out a controlled experiment to test other factors that affect the formation of coacervates. What is included in this resource. Editable lab handouts that are ready to be printed and passed out to your students. Two versions of student handouts: Do you want the students to write their own lab reports, or do you want to provide handouts for students to fill as they work? 5-Page Handout for the Student Designed Experiment. Characteristics of Life PowerPoint (11 slides) Complete instructions. Teacher Guide (6 Pages) containing tips, tricks, and suggestions. Complete Answer Key and Sample Data. Everything you need for the successful completion of this lab. This lab makes an excellent addition to your unit on the origin of life and the early history of life on Earth. I use this lab after teaching this material: The History of Life on Earth PowerPoint With Notes Purpose of the Lab: To simulate Oparin’s work by mixing a protein solution with a carbohydrate solution to produce coacervates. To observe the shared characteristics between coacervates and living cells. To determine the ideal pH level for producing the most coacervates in a solution. To design and carry out a controlled experiment to test other factors that affect coacervate formation. Materials and supplies required for this lab include: Glass test tube with cap, Graduated cylinders, Gelatin solution (protein), Gum Arabic (carbohydrate), Glass stirring rod, pH paper,.1M HCl solution, Dropping pipets, Microscope, Test tube rack, Microscope slides. Included in the download are two versions of the guided lab activity. Use Version 1 (3 pages) of the lab if you want your students to use their own notebook paper to write their own lab reports. Use Version 2 (6 pages) of the lab if you want to print the student data handouts and allow the students to record their data and answers on these handouts. http://crowngreenenergy.com/upload/bt-8568-digital-phone-manual.xml The “student designed experiment” (5 pages) is optional, but highly recommended. This is a time consuming activity, but it is well worth the time to teach your students how to design and carry out a lab experiment. In the handouts provided for the student designed experiment, 6 options are given for additional variables that can be tested. I assign one of these variables to each group of students. You may think of additional testable options that you would like to include. At the conclusion of the lab, I have each group make a short report to the class, giving their hypothesis and the results they obtained. All documents and the PowerPoint are included in multiple formats (doc, docx, PDF, ppt and pptx). Approximate time required to complete: Teacher preparation: 30 minutes Student lab activity: 45 minutes. Students may have to complete some questions for homework. Student designed experiment: 30 minutes for planning and 45 minutes for conducting lab and completing handouts. Report this resource to let us know if this resource violates TpT’s content guidelines. Standards Log in to see state-specific standards (only available in the US). CCSS RST.11-12.3 Follow precisely a complex multistep procedure when carrying out experiments, taking measurements, or performing technical tasks; analyze the specific results based on explanations in the text. CCSS RST.9-10.3 Follow precisely a complex multistep procedure when carrying out experiments, taking measurements, or performing technical tasks, attending to special cases or exceptions defined in the text. Are you getting the free resources, updates, and special offers we send out every week in our teacher newsletter? Sign Up. And I liked it so much that I will definitely use it again next year. Each school year I make myself try new and different activities.Coacervates are simply droplets that are composed ofThis activity was a thought-provoking one for my students. http://www.raumboerse-luzern.ch/mieten/bosch-maxx-wfl-1200-user-manual They clearly were able to see, using a microscope, how it is possible for molecules to come together to form cell-like structures. The first part of my lab was a guided lab activity. The students followed a lab procedure that I gave them to test the effect of different pH levels on the formation of coacervates. But the second day of the lab packed a much bigger punch!! I assigned each lab group a different variable to test. The group had to form a hypothesis and then design a lab to test their hypothesis. For example, some groups had to test the effect of temperature on coacervate formation, while other groups tested the effect of changing solution concentrations on the formation of coacervates. At the conclusion of their experiment, each student group had to make a very short report to the class discussing the results they obtained. This is a time consuming activity, but it is time well spent. Common Core standards are demanding that science students spend more time in analysis, problem solving activities. I required them to form a hypothesis, write a procedure that would gather quantitative data, design a control for the experiment, describe their experimental and control groups, graph their data, form a conclusion, and much more. Arabic (carbohydrate), GlassHCl solution, Dropping pipets, Microscope, TestPros include: Students completed the guided activity in one class period, they wrote and designed their experiment in a second class period, and they carried out their experiment in a third lab period. Some parts of the lab report had to be completed as a homework assignment. The student has to scan the entire slide and count the number of coacervates that were formed. This is a source of error. Share to Twitter Share to Facebook Share to Pinterest I just love getting kids hooked on science!We all know this story. We have lived this story over and over and over in. What will you do to make the first day of school a great day. https://academyocgc.com/images/brinkmann-pro-series-8300-owner-s-manual.pdf What is the most important day of school?. What is paper chromatography. It is a method of sep. This flame test lab is always a favorite of min. When the students go home at the end of. Does this sound familiar. It is nearing the end of the school year, standardized testing is. Sometimes called protocells, these coacervates mimic life by creating vacuoles and movement. All it takes to create these coacervates is protein, carbohydrates, and an adjusted pH. This is easily done in the lab and then the coacervates can be studied under a microscope to observe their life-like properties.Gelatin can be purchased at either the grocery store or a science supply company. Gum acacia is very affordable and can be bought from some science supply companies.There is acid used in this lab, so extra precautions should be taken when working with the chemicals. Make sure the microscope slide and coverslip are clean and ready for use. Fill up the culture tube about half way with the coacervate mix which is a combination of 5 parts gelatin (a protein) to 3 parts gum acacia (a carbohydrate). If this is done properly, it will turn somewhat cloudy. If the cloudiness disappears, add another drop of acid and invert the tube once again to mix. Continue adding drops of acid until the cloudiness stays. Most likely, this will not take more than 3 drops. If it takes more than that, check to be sure you have the right concentration of acid. When it stays cloudy, check the pH by putting a drop on pH paper and record the pH. Cover the mix with a coverslip, and search under low power for your sample. It should look like clear, round bubbles with smaller bubbles inside. If you are having trouble finding your coacervates, try adjusting the light of the microscope. Draw a typical coacervate. Take a drop of the new mix and test its pH by putting it on the pH paper. Follow all safety procedures for working with acid when cleaning. http://hoffmanowska.pl/wp-content/plugins/formcraft/file-upload/server/content/files/16288c028db615---cannondale-jekyll-400-manual.pdf What does this tell you about the acidity of the ancient oceans (if it is assumed this is how life formed)? Hypothesize how you could get the original coacervates to come back into your solution. Create a controlled experiment to test your hypothesis. It looks like your browser needs updating. For the best experience on Quizlet, please update your browser. Learn More What is the Miller Urey model.They can absorb nutrient, grow under the right environment, contain organic molecules. What are some shared characteristics between coacervates and living organisms? protein What type of organic molecule is gelatin. We can't connect to the server for this app or website at this time. There might be too much traffic or a configuration error. Try again later, or contact the app or website owner. Coacervates play an indispensable role in regulating intracellular biochemistry, and their dysfunction is associated with several diseases. Understanding of the LLPS dynamics would greatly benefit from controlled in vitro assays that mimic cells. Protein-pore-mediated permeation of small molecules into liposomes triggers LLPS passively or via active mechanisms like enzymatic polymerization of nucleic acids. This coacervate-in-liposome platform provides a versatile tool to understand intracellular phase behavior, and these hybrid systems will allow engineering complex pathways to reconstitute cellular functions and facilitate bottom-up creation of synthetic cells. These organelles are ubiquitous inside eukaryotic cells, both in the nucleus as well as in the cytoplasm, a few prominent examples being the nucleolus, germ granules, and stress granules 2, 3, 4, 5. One of the most commonly encountered types of condensates are complex coacervates, which form through the electrostatic interaction between oppositely charged polyelectrolytes. cousinsconstructionservices.com/app/webroot/files/calorex-1210-manual.pdf Biomolecules can specifically partition and concentrate inside or at the interface of these compartments, depending on molecular interactions and properties such as solubility, hydrophobic stabilization, and electrostatic complementarity 10, 11, 12, 13. The presence of membraneless compartments in cells have sprouted numerous in vitro studies to further understand their properties and dynamics. For example, enzymatic reaction rates have been observed to strongly increase within coacervates, as recently shown for transcription and translation 14. Additionally, regulation in the form of initiation and dissolution of coacervation has been demonstrated through externally driven enzymatic reactions 15, 16. However, capturing the onset of coacervation and following the subsequent dynamics (nucleation, growth, dissolution, associated chemical reactions, etc.) with high spatiotemporal resolution has found to be challenging in conventional bulk experiments. Indeed, the possibility to induce and limit the coacervation process within a defined volume through external control is highly desirable for in vitro studies. Here, we explore the use of liposomes as controllable containers for in vitro studies of coacervation. Liposomes are compartments consisting of selectively permeable phospholipid bilayers, similar to those found in living cells in the form of cell membrane as well as various intracellular organelles including mitochondria, plastids, endoplasmic reticulum, and secretory vesicles. Liposomes are ideal candidates for serving as bio-compatible micro-environments, since one can encapsulate biomolecules in their interior and membrane proteins (e.g., membrane pores) within the bilayer. However, this system did not take advantage of the functional benefits of membranous structures, such as their selective permeability due to active or passive protein pores. https://aliancegroup.su/wp-content/plugins/formcraft/file-upload/server/content/files/16288c03436cd7---Cannondale-jekyll-4-manual.pdf One feasible approach is to encapsulate one of the essential coacervate components ( C 1 ) inside the liposome and embed protein nanopores in the lipid bilayer. This will allow the size-selective diffusion-mediated transport of remaining components ( C 2 ) into the liposomal lumen, thereby providing external control over the onset and process of coacervate formation. We form the liposomes using recently developed on-chip microfluidic method, Octanol-assisted Liposome Assembly (OLA) 18. Addition of further components in the external environment allows passive transport into the lumen, thereby commencing the process of coacervate formation. We furthermore show the potential functionality of the coacervates as rudimentary membraneless organelles by sequestering protein molecules (FtsZ, a key protein for bacterial cell division) as well as supramolecular assemblies such as small unilamellar vesicles (SUVs, a lipid source to potentially achieve liposome growth and form membranous sub-compartments). Summing up, our liposome-based platform provides spatiotemporal control to study the process of forming functional coacervates in sub-picoliter confinements. Fig. 1 Controlled formation of membraneless coacervates in liposomes.By inserting bilayer-spanning protein pores, one can allow passive transport of small molecules, leading to the formation of a coacervate. Such a coacervate-in-liposome hybrid system can also be used as a scaffold for a synthetic cell, where the liposome represents the primary compartment (a cell), while the coacervate represents a sub-compartment (an organelle).Liposomes encapsulating one of the coacervate component ( C 1 ) or an enzyme catalyzing the production of C 1 are generated using OLA. The presence of high-density molecules, such as dextran, efficiently settles them at the bottom of the collection well, while the waste products (1-octanol droplets) float to the top. http://andreagarciam.com/wp-content/plugins/formcraft/file-upload/server/content/files/16288c03a2d883---cannondale-gemini-dh-manual.pdf The other coacervate component, C 2, can already be present in the chamber or can be added later to induce coacervation.The red boundary indicates the lipid bilayer while the green lumen shows encapsulated FITC-pLL molecules.The slight offset between pLL and ATP fluorescence is due to the diffusion of the coacervate between capturing of the images. Absorbance of three independent samples was measured (nine measurements per sample).Error bars in ( f, g, i ) indicate standard deviations. Source data are provided as a Source Data file Full size image The described methodology to build hybrid systems comprised of membranous and non-membranous scaffolds also opens further avenues to increase the complexity in bottom-up synthetic biology, where one of the major goals is to establish a synthetic cell from molecular components. Hybrid coacervate-in-liposome systems can potentially be endowed with unique properties that arise due to the complementarity of their constituents 19, 20, 21. While liposomes can effectively encapsulate molecules and form transmembrane gradients, coacervates enable a local heterogenous increase of charged and hydrophilic molecules 22. Thus, their combination may allow for the creation of synthetic cells with a degree of heterogeneity that typically is observed in living cells. Results On-chip experimental set-up to study coacervate dynamics We set out to induce the controlled formation of membraneless coacervates inside liposomes (Fig. 1a ). The idea was to encapsulate part of the necessary components ( C 1 ) inside the liposomes and allow the transport of the remaining necessary component ( C 2 ) through protein pores in the membrane. We separated the formed liposomes from the waste product (less dense 1-octanol droplets) using a modified version of a density-based separation technique that we have previously reported 23, 24. We implemented two major changes (Fig. www.corwell.co.uk/userfiles/files/calore-space-heater-manual.pdf 1b ): Firstly, we punched a large collection well at the end of the production channel in order to collect liposomes at its bottom and let the waste product (1-octanol) float to the top of the buffer-filled well. This improvement of encapsulating dense molecules to induce the settling of liposomes, led to their complete isolation from unwanted side-products of OLA and allowed straightforward long-term experimentation. Colocalization of the ATP signal with that of the pLL in the condensed phase clearly proved that the diffusion of ATP through the membrane pores led to the formation of coacervates (Fig. 1e ). We also performed a bulk turbidity assay to estimate the ATP concentration above which coacervation took place (see Methods). Consequently, as soon as the diffusive transport across the porous membrane increased the internal ATP concentration above this threshold, coacervation took place. The microfluidic set-up allowed us to obtain monodisperse coacervate-containing liposome samples. Indeed, the corresponding coefficients of variation of respectively 6 and 11 indicate a low degree of variability. We tested this by varying the pLL concentration inside the liposomes and measuring the size of the formed coacervates (see Methods for details). As can be seen in Fig. 1g, the mean coacervate diameter indeed increased with the amount of encapsulated pLL; cf. We tested this using fluorescently labeled dextran (AF647-dextran). The results showed no significant sequestration of dextran molecules inside the coacervates but instead showed a similar fluorescence intensity as in the rest of the liposomal lumen, with a slight accumulation at the coacervate interface (Supplementary Fig. 2 ). This observed accumulation at the interface clearly did not alter the coacervation process, as the coacervation dynamics remained unchanged, independent of whether we used sucrose or dextran to settle the liposomes. We also note that technical problems in liposome production, such as an undesirable bursting of double-emulsion droplets, can release some encapsulated coacervate components (e.g., pLL) into the collection well, leading to a residual amount of unwanted coacervation outside the liposomes, as can for example be noted in Fig. 1e. A detailed troubleshooting to ensure stable liposome production can be found in our online protocol 24. Our approach enables to measure the time dependence of the coacervation process. We captured time-lapse movies by gently introducing the ATP-containing solution into the well after the liposomes were settled to the bottom. The process is shown schematically in Fig. 2a, and the time-lapse images, visualized by the fluorescence of FITC-pLL, are shown in Fig. 2b (also see Supplementary Movie 1 ). ATP molecules rapidly diffused throughout the well, and entered the liposomes through the pores. Once the threshold ATP concentration required for coacervation was reached inside the vesicle, a rapid phase transition was observed. Small, discrete condensates appeared throughout the liposomal lumen, with a bright fluorescence indicating a high concentration of pLL molecules inside them. It interacts with pLL molecules present inside the liposome, initiating coacervation throughout the liposome. Over time, individual coacervates coalesce to form a single coacervate.Note the simultaneous formation of multiple coacervates that further coalesce to form one single entity. Time zero ( t 0 ) corresponds to the first visible sign of coacervation.The rapid transition from a homogeneous solution to a condensed phase leads to sudden rise in the fluorescence intensity, which plateaus over time. The horizontal dashed lines indicate the decrease in the fluorescence intensity just before coacervation takes place, providing evidence for nucleation events. Error bars in ( d, e ) indicate standard deviations. Dashed vertical lines in ( f, g ) indicate the onset of coacervation, the plots show the average values, with the shaded regions indicating standard deviations. Next, we attempted to detect the nucleation events that led to the formation of condensates by plotting the dilute-phase intensity over time, a quantity which corresponds to the less-bright fluorescence background intensity inside the liposomes. We observed a rapid decay, complementary with the formation of the coacervate phase (Fig. 2g ). This is expected, as coacervation considerably decreases the fraction of pLL molecules that reside in the dilute phase, decreasing the background intensity in the liposomal lumen. We speculate that this fluorescence intensity loss, just before the coacervates are observed, indicates the nucleation process that precedes the observable formation of coacervates. The observed changes in the fluorescence intensity were not a result of photobleaching, as the total fluorescence counts of a liposome not showing coacervation stayed constant over a similar period of time (Supplementary Fig. 3 ). Along with capturing the coacervation dynamics, we simultaneously assessed the continuous interaction of the formed coacervate with the surrounding dilute phase. We did this by encapsulating apyrase inside the liposome, an enzyme that degrades ATP into adenosine diphosphate (ADP) and finally into adenosine monophosphate (AMP). In contrast, the coacervates inside the liposomes remained highly stable in the presence of apyrase, even over a course of hours (Supplementary Movie 3 ). This showed that there was a constant exchange of coacervate material with the environment, i.e., ATP was continuously replenished. Note that in the absence of apyrase, the coacervation progressed in an exactly similar fashion, both in terms of the dynamics and the time scale (Supplementary Fig. 5 ). Clearly, the degradation activity of apyrase was not strong enough to counterbalance the influx of ATP through the pores and thus did not affect the induction of coacervation. As a side note, the onset and subsequent progression of coacervation seen in a minor fraction of leaky liposomes, in the absence of membrane pores, was similar to that seen in pore-containing liposomes (Supplementary Fig. 6 ). Overall, we conclude that pore-permeated liposomes present a viable strategy to form stable coacervates in a controlled manner. UDP was provided externally, and spermine was present inside and outside of the vesicle, serving as a positively charged polyelectrolyte. Note the simultaneous formation of several coacervates that further coalesce to form one single entity.The transition from a homogeneous solution into a condensed coacervate phase leads to a gradual increase in the fluorescence. As the coacervates continue to grow, there is a gradual, linear decrease in the fluorescence of the dilute phase. Error bars in ( c, d ) indicate standard deviations. Dashed vertical lines in ( e, f ) indicate the onset of coacervation, the plots show the average values, with the shaded regions indicating standard deviations. We observed that the coacervates remained stable and did not increase in size even after a few hours (Supplementary Movie 5, Supplementary Fig. 7 ). With virtually unlimited supply of spermine and UDP, one would expect the coacervates to continuously grow. However, a limited polymerization activity of PNPase and activation of the RNA exoribonuclease activity of the enzyme 33 possibly inhibited further growth. The fluorescence intensity remained relatively constant after the initial increase. We focused on two salient and well-known features of condensates that bring out their potential to act as membraneless organelles: the specific sequestration of biomolecules and their use as reaction centers 6, 10, 12, 14, 34. First, we encapsulated FtsZ protein inside the liposomes. FtsZ is a bacterial protein crucially involved in the process of cell division 35. We observed that FtsZ sequestration occurred in a highly efficient way. In the current case, however, FtsZ was already present when the phase separation occurred. We observed a similar homogenous distribution of FtsZ inside the coacervates, when the coacervates were prepared in the absence of any vesicles (Supplementary Fig. 8 ). This ruled out any effect of the membranous confinement on the spatial organization of FtsZ within the coacervate. These experiments suggest that the spatial organization of sequestered molecules within the coacervate depends on the sequence and timing of the addition of parts. Fig. 4 Sequestration of biomolecules and compartmentalization of reactions within coacervates.The circles represent the individual data points, bars represent the mean fluorescence intensity, and the error bars indicate corresponding standard deviations. A similar homogenous SUV distribution was obtained, when the coacervates were prepared in the absence of any vesicles, ruling out any effect by the compartmentalization (Supplementary Fig. 9 ). Our experiments with FtsZ and SUVs suggest that the sequence of reactions plays an important role in determining the nature of sequestration. Finally, we investigated the possibility to carry out a biochemical reaction specifically inside the coacervates. The reaction product (fluorescein) likely remained confined within the condensed phase due to hydrophobic stabilization and potential electrostatic interactions with cationic groups of pLL 12. Summing up, we successfully demonstrated two important attributes of coacervates: sequestering and concentrating biomolecules, and serving as hubs for biochemical reactions. Discussion In this paper, we reported the controlled formation of hybrid microcontainers, viz., coacervates-in-liposomes. We show that these are ideal systems to study the dynamics of LLPS, and are also well suited as potential architectural scaffolds for the design of future synthetic cells. We employed the on-chip microfluidic method OLA 18 to produce liposomes, where a collection well at the end of the production channel allowed to immediately settle and visualize the liposomes, a process facilitated by making the liposomes denser by encapsulating sucrose or dextran. With this platform at hand, we obtained temporal control over the onset of coacervation as well as the final size of condensates. The temporal control was obtained by the addition of essential components for condensation from the outside, and their entry into the liposomes through protein nanopores present in the lipid bilayer, in order to trigger LLPS. Control over the condensate size was achieved by encapsulating a precise and finite amount of material within the liposome. The ability to induce and selectively limit the coacervation process presents a crucial advantage for experiments in which spatiotemporal control and monitoring is desirable. For example, using fluorescence intensity analyses, we were able to detect the de novo nucleation of condensates in real time, a process which has been difficult to study and control so far 1, 36. We were further able to follow and compare the dynamics of two different coacervate systems to demonstrate the strength of our experimental setting: (i) A straightforward coacervation reaction mediated through diffusive transport of a necessary component across the membrane. (ii) A more complex scenario where real-time production of a multivalent polymer subsequently triggered the coacervation process. We analyzed and quantified the differences between the two systems, in terms of the evolution of the condensed and the dilute phase, as well as the number and size of the formed coacervates.