The scarred original position: Re-testing foundational political theory after collapse

This study demonstrated the theory of Thomas Hobbes on the necessity to move from the state of nature and form a state. Law is a prerequisite for ensuring peace, tranquillity and effective governance in a society. When law is in place, it fosters human beings' interaction with each other and creates a conducive environment for socio-economic empowerment. In the state of nature, according to Thomas Hobbes, there existed no government, no law, and nobody was above the other. There was no law, industry, civilisation, social life, economic and political order. If there was no law in place, then there was no morality. If there was no morality, then there was no justice or injustice. Life here was characterised by brutality, selfishness, and solitude and hence was short. The study was guided by Thomas Hobbes's political theory, which emphasises consent and consensus in the formation of government so as to move from the state of nature where life was unbearable. The study was library-based, and the researcher used a purposive sampling method to purposely sample three books that are inclined to materials related to Thomas Hobbes's political theory on the state of nature. The data was collected through reading texts, both published and unpublished. The location of the study was Chuka University Library. The study adopted a descriptive design and used a critical method that questions assumptions, examines the evidence, identifies biases, and strives for a deeper understanding of the state of nature and the necessity of forming a society. The study found that there is a need to have an organised society guided by law. The study recommended the need for equal distribution of resources.

Political philosophers often draw their conclusions on how political systems ought to be by first investigating human nature and then proposing recommendations extrapolating from those investigations. They attempt to do this by creating a hypothetical ‘state of nature’ where human beings would be unaffected by social, political, and cultural paradigms and can act freely in pursuit of their instincts, thereby revealing their true nature. English philosopher Thomas Hobbes followed this method of investigation and found that human beings are naturally violent, individualistic, and warlike and therefore concluded that political theory ought to be as such to prevent human beings from acting out their baser impulses through draconian authoritative measures when necessary. In this paper, I challenge Hobbes’s claim on human nature by citing contemporary research on cognitive sciences which demonstrates that human beings have a natural predisposition to believe in supernatural agency (God/s) and therefore in the state of nature must behave in such a way as contemporary psychologists discovered investigating the effect of religiosity on human beings such as increased cooperation, selfless behavior towards genetically unrelated strangers, reduced crime rate deterred by supernatural punishment, etcetera. I argue that if we take into account the human propensity to believe in the supernatural agency then the state of nature hypnotized by Hobbes would need to be modified and therefore so would his political theory. Given the integration of cognitive science of religion into the state of nature, I conclude that the ideal political system must be centered around religion. Philosophy and Progress, Vol#67-68; No#1-2; Jan-Dec 2020 P 77-118

The concept of State of Nature is important in political theory because it helps us understand the origin of political authority and the role of government in society. The State of Nature refers to the hypothetical condition of human beings before or without political association. The analysis is about the comparison of Islamic and western views by using two figures, Al-Mawardi and Thomas Hobbes. This research aims to compare their opinion regarding the State of Nature from the perspective of Siyasah Syar’iyyah. This research uses a comparative approach and content analysis. This study discusses the critical analysis of Thomas Hobbes and Al-Mawardi's views on the concept of a state of nature. Thomas Hobbes conducted that the state of nature is the basic nature of humans who are suspicious of each other at war to find happiness or Hobbes calls it Homo humini lupus, which mean the state of the werewolf for other humans, in contrast to Al-Mawardi's view that humans are deliberately created as weak creatures so that they need interaction to help each other, humans are social creatures who need other people to survive. The research results show that Al-Mawardi and Thomas Hobbes have different views about the State of Nature. According to Al-Mawardi, humans are born in a good natural state, whereas according to Thomas Hobbes, humans are born in a bad natural state. Apart from that, Al-Mawardi argued that humans have rights that must be respected by the state, while Thomas Hobbes argued that humans must surrender their rights to the state for the sake of security and peace.

The Analysis of the Relationship between God, Religion and Politics in Thomas Hobbes’s Leviathan and De Cive

Article Figures and data Abstract Editor's evaluation eLife digest Introduction Results Discussion Materials and methods Data availability References Decision letter Author response Article and author information Metrics Abstract Fixing cells with paraformaldehyde (PFA) is an essential step in numerous biological techniques as it is thought to preserve a snapshot of biomolecular transactions in living cells. Fixed-cell imaging techniques such as immunofluorescence have been widely used to detect liquid–liquid phase separation (LLPS) in vivo. Here, we compared images, before and after fixation, of cells expressing intrinsically disordered proteins that are able to undergo LLPS. Surprisingly, we found that PFA fixation can both enhance and diminish putative LLPS behaviors. For specific proteins, fixation can even cause their droplet-like puncta to artificially appear in cells that do not have any detectable puncta in the live condition. Fixing cells in the presence of glycine, a molecule that modulates fixation rates, can reverse the fixation effect from enhancing to diminishing LLPS appearance. We further established a kinetic model of fixation in the context of dynamic protein–protein interactions. Simulations based on the model suggest that protein localization in fixed cells depends on an intricate balance of protein–protein interaction dynamics, the overall rate of fixation, and notably, the difference between fixation rates of different proteins. Consistent with simulations, live-cell single-molecule imaging experiments showed that a fast overall rate of fixation relative to protein–protein interaction dynamics can minimize fixation artifacts. Our work reveals that PFA fixation changes the appearance of LLPS from living cells, presents a caveat in studying LLPS using fixation-based methods, and suggests a mechanism underlying the fixation artifact. Editor's evaluation Chemically fixing cells for fluorescence microscopy is a common practice in cell biology. However, fixation artifacts can lead the incorrect interpretations of experimental results. This article presents compelling evidence showing that in the context of liquid condensates formed by liquid–liquid phase separation (LLPS), paraformaldehyde (PFA) fixation creates a number of artifacts – such as changes in the number, appearance, or disappearance of liquid condensates. These important findings will be of great interest not only for those in the LLPS field but for any cell biologists using fixed samples for microscopy. https://doi.org/10.7554/eLife.79903.sa0 Decision letter Reviews on Sciety eLife's review process eLife digest A typical human cell is a crowded soup of thousands of different proteins. One way that the cell organizes this complex mix of contents is by creating separate droplets within the cell, like oil in water. These droplets can form through a process known as liquid-liquid phase separation, or LLPS, where specific proteins gather in high concentrations to carry out their cellular roles. The critical role of LLPS in cellular organization means that it is widely studied by biologists. To detect LLPS, researchers often subject the cells to treatments designed to hold all the proteins in place, creating a snapshot of their natural state. This process, known as fixing, allows scientists to easily label a protein with a fluorescent tag, take pictures of the cells, and look at whether the protein forms droplets in its natural state. This is often easier to do than imaging cells live, but it relies on LLPS being well-preserved upon fixation. To test if this is true, Irgen-Gioro, Yoshida et al. looked at protein droplets in live cells, and then fixed the cells to check whether the appearance of the droplets had changed. The images taken showed that fixation could alter the size and number of droplets depending on the protein being studied. To explain why the effects of fixing change depending on the protein, Irgen-Gioro, Yoshida et al. hypothesized that a faster fixation – relative to how quickly proteins can bind and unbind to their droplets – can better preserve the LLPS droplets. They verified their idea using a microscopy technique in which they imaged single molecules, allowing them to see how different fixation speeds relative to protein binding affected the droplets. The work of Irgen-Gioro, Yoshida et al. identifies an important caveat to using fixation for the study of LLPS in cells. Their findings suggest that researchers should be cautious when interpreting the results of such studies. Given that LLPS in cells is an area of research with a lot of interest, these results could benefit a broad range of biological and medical fields. In the future, Irgen-Gioro, Yoshida et al.’s findings could prompt scientists to develop new fixing methods that better preserve LLPS in cells. Introduction Fixing cells to preserve a snapshot of biomolecular transactions in vivo is a widely used strategy in numerous techniques in biology and medicine. Due to its small size and high reactivity with a wide range of biological entities, paraformaldehyde (PFA) is one of the most commonly used fixatives to create covalent cross-linking between biomolecules, for example, proteins and nucleic acids. PFA nonselectively ‘fixes’ or cross-links molecules in proximity to enable characterization of biomolecular interactions formed in living cells. Examples of popular techniques that use PFA to fix cells include ChIP-sequencing (Robertson et al., 2007; Solomon and Varshavsky, 1985), chromosome conformation capture (3C)-based techniques (Dekker et al., 2002), immunofluorescence (Richter et al., 2018), fluorescence in situ hybridization (FISH) (Moter and Göbel, 2000), cross-linking mass spectrometry (Sutherland et al., 2008), super-resolution expansion microscopy (Chen et al., 2015), and super-resolution localization microscopies such as stochastic optical reconstruction microscopy (STORM) (Rust et al., 2006). Although PFA fixation has been used to faithfully preserve live-cell conditions in many scenarios, a number of studies have uncovered situations in which fixation fails to cross-link DNA–protein interactions formed in living cells. By imaging different transcription factors (TFs) in live and fixed cells, Schmiedeberg et al., 2009 showed that TFs bound to DNA with fast dissociation dynamics (<5 s residence times as determined by fluorescence recovery after photobleaching [FRAP]) are not cross-linked to DNA upon PFA fixation. Using live-cell single-molecule imaging, Teves et al., 2016 showed that TFs stay bound to chromosome during mitosis and fixing cells can artificially deplete transiently bound TFs from mitotic chromosomes. These studies exemplify the fact that fixation, with limited reaction rates, cannot provide an instantaneous snapshot and may miss or obfuscate biomolecular interactions that happen either at or faster than the timescale of fixation. What further complicates the result of cell fixation is that the reactivity and reaction rates of PFA are variable and dependent on its biomolecule substrates (Gavrilov et al., 2015; Shishodia et al., 2018). For example, the efficiency and rates at which PFA reacts with proteins can vary by orders of magnitude (Kamps et al., 2019) and are dependent on their amino acid sequences (Kamps et al., 2019; Metz et al., 2004; Sutherland et al., 2008) and tertiary structures (Hoffman et al., 2015). Among the numerous biomolecular transactions investigated using fixed-cell imaging is liquid–liquid phase separation (LLPS), a long-observed behavior of polymers in solution (Gibbs, 1879; Graham, 1861; Hyman et al., 2014) that has recently generated much excitement in biological research communities due to its proposed roles in cellular organization and functions (Banani et al., 2017; Boeynaems et al., 2018; Mitrea and Kriwacki, 2016; Shin and Brangwynne, 2017). LLPS is driven by excessive levels of transient, selective, and multivalent protein–protein interactions mediated by intrinsically disordered regions (IDRs) within the proteins of interest (Chong et al., 2018; Kato and McKnight, 2018; Li et al., 2012). Whereas rigorous characterization of LLPS in vivo has been challenging and remains a question under active investigation (McSwiggen et al., 2019b), detection of discrete puncta that have a spherical shape, undergo fusion and fission, and dynamically exchange biomolecules with the surrounding according to FRAP is often considered evidence of putative LLPS in living cells. While such diverse measurements have been widely used for studying proteins under overexpression conditions, far fewer approaches are available to probe LLPS under physiological conditions. Detecting local high-concentration regions or puncta of an endogenously expressed protein using immunofluorescence of fixed cells has been used in many studies as evidence of LLPS (Boija et al., 2018; Guo et al., 2019; Owen et al., 2021; Xie et al., 2022; Yang et al., 2020). Not only is the detection of puncta an inconclusive metric for establishing LLPS, whether a punctate distribution observed in fixed cells actually represents the live-cell scenario remains unclear as fixation has only been assumed, but not directly shown, to faithfully preserve multivalent interactions and LLPS formed in living cells. This knowledge gap motivated us to image cells that overexpress various known IDR-containing proteins before and after fixation to evaluate the ability of PFA fixation to preserve LLPS behaviors. We found that, interestingly, fixation can significantly alter the appearance of droplet-like puncta in cells. Our quantitative image analysis suggests that depending on the LLPS-driving protein, fixing cells can either enhance or diminish the apparent LLPS behaviors in vivo. In certain cases, fixation can even cause droplet-like puncta to artificially appear in cells that have a homogeneous protein distribution and no detectable puncta in the live condition. Conversely, fixation can also cause droplet-like puncta in living cells to completely disappear. Combining experiments that modulate fixation rates, live-cell single-molecule imaging that quantifies protein binding dynamics, and simulations based on a kinetic model, we further demonstrated that protein localization in fixed cells depends on an intricate balance of protein–protein interaction dynamics, the overall rate of fixation, and the difference between protein fixation rates in and out of droplet-like puncta. Our work urges caution in the interpretation of previous claims of in vivo phase separation based solely on immunofluorescence imaging of fixed cells and serves to guide future judicious application of PFA fixation. Results Fixation enhances the LLPS appearance of FET family proteins To investigate the effect of PFA fixation on the appearance of LLPS, we first compared confocal fluorescence images of live and fixed U2OS cells that transiently express an IDR tagged with EGFP and a nuclear localization sequence (NLS). We focused on the FET family protein IDRs (AA2-214 of FUS, AA47-266 of EWS, and AA2-205 of TAF15) that are reported to undergo putative LLPS in cells upon overexpression (Altmeyer et al., 2015; Chong et al., 2018; Wang et al., 2018). Figure 1, Video 1, and Figure 1—figure supplement 1 compare the same cells before and after treatment of 4% PFA for 10 min unless otherwise noted, a typical condition utilized for fixed-cell imaging techniques such as immunofluorescence. At high enough expression levels, all three IDRs are able to form discrete and spherical puncta in the live cell nucleus, which show fusion and fission behaviors and are thereby consistent with LLPS droplets (Alberti et al., 2019; Banani et al., 2017). Interestingly, after fixation, the puncta of all three IDRs appear to increase in their numbers, sizes, and contrast compared with the dilute phase. In particular, PFA fixation was able to artificially turn a cell with EGFP-EWS(IDR) homogeneously distributed in the nucleus without any puncta into one with many discrete puncta (Figure 1). We quantified the fixation-induced changes of LLPS appearance by calculating three parameters from the fluorescence images of cells, including the number of puncta, surface roughness, and punctate percentage, and found a significant increase in all three parameters after fixation (Figure 1D–F, Figure 1—source data 1). The number of puncta and punctate percentage (percentage of intranuclear fluorescence intensity in the concentrated phase) are indicators of the propensity to phase separate (Berry et al., 2015). The surface roughness (standard deviation of pixel intensities across the nucleus) quantifies the uneven distribution of a fluorescently labeled protein in the nucleus, allowing for detection of puncta appearance or disappearance without the need for an algorithm to identify individual puncta in the cell. Figure 1 with 4 supplements see all Download asset Open asset Fixation can change the apparent liquid–liquid phase separation (LLPS) behaviors of proteins. (A) EGFP-EWS(IDR), (B) EGFP-FUS(IDR), and (C) EGFP-TAF15(IDR) are transiently expressed in U2OS cells and imaged before and after fixation using confocal fluorescence microscopy. A schematic of each protein construct is shown on the left. A maximum z-projection of a representative live cell expressing its respective protein is shown next to that of the same cell after 10 min of fixation with 4% paraformaldehyde (PFA). The inserts show a zoomed-in region of the cell. (D–F) Quantification of percentage change of LLPS parameters after fixation. The values are averaged from 34 (D), 17 (E), or 24 (F) cells measured in 3 (D), 2 (E), or 2 (F) independent transfection and imaging sessions. Error bars represent standard errors. Asterisks indicate a significant difference compared with 0 (p<0.05, Wilcoxon signed-rank test). Figure 1—source data 1 Quantification of puncta parameters used to generate the bar plots. https://cdn.elifesciences.org/articles/79903/elife-79903-fig1-data1-v3.xlsx Download elife-79903-fig1-data1-v3.xlsx Video 1 Download asset This video cannot be played in place because your browser does support HTML5 video. You may still download the video for offline viewing. Download as MPEG-4 Download as WebM Download as Ogg Real-time imaging of a U2OS cell expressing EGFP-FUS(IDR) during paraformaldehyde (PFA) fixation. We next tested how the fixation artifact is dependent on the length of PFA treatment, PFA concentration, and the type of fixatives. We performed real-time imaging of live cells expressing EGFP-FUS(IDR) and found that their morphology and LLPS appearance start to change immediately upon PFA treatment and reach a steady state after ~100 s of treatment (Video 1, Figure 1—figure supplement 2). We treated cells expressing EGFP-EWS(IDR) with different concentrations of PFA (1, 2, 4, and 8%) and observed statistically significant changes to the above three LLPS-describing parameters upon fixation at all the concentrations (Figure 1—figure supplement 3). PFA in combination with glutaraldehyde (GA) has been shown to reduce fixation artifacts in imaging the distribution of cell membrane receptors (Stanly et al., 2016). However, we still observed statistically significant fixation-induced changes to the apparent LLPS behavior of EGFP-EWS(IDR) using 4% PFA and 0.2% GA in combination (Figure 1—figure supplement 4). We next compared the intracellular distribution of TAF15(IDR) tagged with different fluorescent tags, including, EGFP, DsRed2, and HaloTag, before and after fixation with 4% PFA. The LLPS behavior of DsRed2-TAF15(IDR) is enhanced upon fixation like EGFP-TAF15(IDR) (Figure 2A), but the enhancement has a different appearance. Whereas there is not a significant change to the large preformed DsRed2-TAF15(IDR) puncta, thousands of smaller puncta emerge in the dilute phase within the nucleus (Figure 2B). In contrast, Halo-TAF15(IDR) displays a diminished LLPS behavior after fixation, with its puncta becoming smaller and dimmer or completely disappearing (Figure 2C, Figure 2—figure supplement 1). Quantification of the number of puncta, surface roughness, and punctate percentage of the TAF15(IDR) LLPS systems before and after fixation further confirmed these observations (Figure 2D–F, Figure 2—source data 1). The fact that different phase-separating proteins can have bifurcating behaviors upon fixation is interesting. While it is known that EGFP and DsRed2 can dimerize and HaloTag cannot (Costantini et al., 2012; Sacchetti et al., 2002), it is unclear whether and how the dimerization potential might contribute to the proteins’ bifurcating responses to PFA fixation. We note that the fixation-induced changes to LLPS appearance can affect the physical characterization of in vivo LLPS systems based on fixed-cell imaging, such as the Gibbs energy of transfer between dilute and concentrated phases (Riback et al., 2020) and how far from the critical concentration a system is (Bracha et al., 2018), potentially affecting the interpretation of the functional role of LLPS in cellular processes. Moreover, the fact that PFA fixation can artificially promote puncta formation even in cells without detectable puncta in the live condition presents an important caveat in fixation-based approaches that have been commonly used for characterizing LLPS under physiological conditions, for example, immunofluorescence. Figure 2 with 1 supplement see all Download asset Open asset Paraformaldehyde (PFA) fixation can both enhance and diminish liquid–liquid phase separation (LLPS) appearance. U2OS cells expressing (A) EGFP-TAF15(IDR), (B) DsRed2-TAF15(IDR), and (C) Halo-TAF15(IDR), ligated with the JFX549 Halo ligand, are imaged using confocal fluorescence microscopy before and after 10 min of fixation with 4% PFA. Schematics of the protein constructs are shown on the left. Live- and fixed-cell images are compared. (D–F) Quantification of LLPS parameters after fixation. The values are averaged from 24 (D), 23 (E), or 10 (F) cells measured in 2 (D), 2 (E), or 3 (F) independent transfection and imaging sessions. Error bars represent standard errors. Asterisks indicate a significant difference compared with 0 (p<0.05, Wilcoxon signed-rank test). Figure 2—source data 1 Quantification of puncta parameters used to generate the bar plots. https://cdn.elifesciences.org/articles/79903/elife-79903-fig2-data1-v3.xlsx Download elife-79903-fig2-data1-v3.xlsx Furthermore, to examine whether all phase-separating proteins show the fixation artifact, we compared live- and fixed-cell images of EGFP-tagged full-length FUS (FUS(FL)). Full-length FUS is reported to have a greater LLPS propensity in vitro than its IDR alone (Wang et al., 2018). We found that EGFP-FUS(FL) overexpressed in live U2OS cells forms many small puncta throughout the nucleus, and we did not observe a significant change of this behavior after PFA fixation (Figure 3A, Figure 3—source data 1). We also fused Halo-tagged TAF15(IDR) to FTH1 that forms a 24-mer (Bellapadrona and Elbaum, 2014 and Bracha et al., 2018) to make an artificial protein with a high LLPS propensity. We found that TAF15(IDR)-Halo-FTH1 overexpressed in live U2OS cells forms large droplet-like puncta and the appearance of LLPS does not significantly change after PFA fixation (Figure 3B, Figure 3—source data 1). In addition, we looked into a native IDR-containing protein, EWS::FLI1, an oncogenic TF causing Ewing sarcoma (Grünewald et al., 2018) and known to form local high-concentration hubs at target genes associated with GGAA microsatellites (Chong et al., 2018). Although there is no convincing evidence that EWS::FLI1 undergoes LLPS under physiological conditions, the formation of its hubs is mediated by the homotypic multivalent interactions of EWS(IDR) within the protein. Excessive levels of such multivalent interactions often result in LLPS (Li et al., 2012). We previously Halo-tagged endogenous EWS::FLI1 in an Ewing sarcoma cell line A673 using CRISPR/Cas9-mediated genome editing (Chong et al., 2018). Here, we compared live and fixed A673 cell images of endogenous EWS::FLI1-Halo and did not observe a significant difference in its distribution (Figure 3C, Figure 3—source data 1). This result suggests that PFA fixation does not change the intracellular distribution of all proteins that have a LLPS potential. Figure 3 Download asset Open asset Not all puncta-forming proteins show the fixation artifact. U2OS cells expressing (A) EGFP-FUS(FL) and (B) TAF15(IDR)-Halo-FTH1, and (C) an A673 cell expressing endogenous EWS::FLI1-Halo are imaged using confocal fluorescence microscopy before and after 10 min of fixation with 4% paraformaldehyde (PFA). Halo-tagged proteins are ligated with the JFX549 Halo ligand before imaging. Schematics of the protein constructs are shown on the left. Live- and fixed-cell images are compared. (D–F) Quantification of puncta parameters after fixation. The values are averaged from 21 (D), 16 (E), or 15 (F) cells measured in 1 (D), 4 (E), or 2 (F) independent transfection and imaging sessions. Error bars represent standard errors. NS: not significant difference compared with 0 (p<0.05, Wilcoxon signed-rank test). None of the examined proteins show significant changes in their liquid–liquid phase separation (LLPS) or hub appearance in the fixed-cell image as compared to the live-cell image. Figure 3—source data 1 Quantification of puncta parameters used to generate the bar plots. https://cdn.elifesciences.org/articles/79903/elife-79903-fig3-data1-v3.xlsx Download elife-79903-fig3-data1-v3.xlsx Switching between enhancing and diminishing the LLPS appearance depends on fixation kinetics To understand what factors are underlying the diverging fixation artifact of in vivo LLPS we performed the fixation imaging with to live cells to PFA fixation. is with and is commonly used to the formation of protein–protein cross-linked by quickly and cross-linked (Hoffman et al., 2015). We utilized to generate a fixation reaction in the cell protein–protein fixation. We found that to live U2OS cells that overexpress DsRed2-TAF15(IDR) the punctate percentage from to from 23 an increase in the of LLPS. Although the underlying mechanism of such increase is we this might be because that an important role in TAF15(IDR) LLPS et al., are enhanced by the presence of of the fixation effect on the LLPS behavior of Whereas PFA fixation in the of enhances the LLPS appearance (Figure Figure in the presence of glycine, fixation many of the smaller puncta formed in live cells to completely and preformed puncta to turn into a shape, with the of the puncta still but the of the protein (Figure None of these fixed-cell images are of live cells, but it that the critical parameters that the artifact of PFA fixation. The that the appearance of droplet-like puncta in fixed cells can be by the presence of that the kinetics of fixation can an essential role in the appearance of LLPS in fixed cells. Figure 4 Download asset Open asset fixation creates a fixation artifact. (A) Fixing U2OS cells that express DsRed2-TAF15(IDR) in the of many small puncta to (B) Fixing cells in the presence of results in a in the number of puncta, with large puncta In both (A) and cells are imaged using confocal fluorescence microscopy before and after 10 min of fixation with 4% paraformaldehyde (PFA). the fixation artifact Given that fixation kinetics are critical to the appearance of LLPS in fixed cells, we a kinetic model et al., 2006). shown in Figure and the model on one protein of interest which before fixation can either be in state – or – of molecules are dynamically in and out of puncta, the percentage of is at an determined by the of the binding and the dissociation These are the exchange rates between and and do not the potential in the rates at the For example, individual molecules at the surface and of a might with different rates, but model does not these We the that PFA is as and fixed of which are cross-linked to proteins within with a fixation rate of and cross-linked to proteins with a fixation rate of fixing to both and are when the cell is fixed after of PFA there is no any concentration in and The fixation artifact of an LLPS system can be as the change in punctate percentage, or the of to after Figure Download asset Open asset bifurcating fixation artifacts. (A) that fixation of a phase-separating protein of interest in the cell. (B) The kinetic model with associated kinetic rates the different (C) of the fixation artifact as a of the punctate percentage and the relative fixation rate the overall fixation rate as as overall protein binding and dissociation rates are fixation liquid–liquid phase separation (LLPS) behavior to be fixation LLPS behavior to be of the fixation artifact as a of the punctate percentage and the relative overall fixation rate individual fixation rates are overall fixation rate compared with protein–protein interaction dynamics the fixation artifact. (C) and punctate from to are on (C) and We hypothesized that the balance between interaction and fixation dynamics in a LLPS system the fixation artifact and tested the by calculating as a of various kinetic and is that the dilute and concentrated phases of an LLPS system have different protein and concentrations and 2022; et al., et al., 2015; et al., The rate of fixation is known to vary with both factors by orders of with the timescale of fixation from to (Hoffman et al., 2015; et al., 2019; Metz et al., Metz et al., protein–protein interactions that LLPS are dynamic with binding residence times in the range of to of (Chong et al., 2018), fixation with either or rates than protein binding and We first examined whether different fixation rates of in and out of puncta can cause a fixation artifact, the overall fixation rates are than protein binding and and how the fixation artifact may on protein–protein interaction we as a of the punctate percentage and the relative fixation rate when the relative overall fixation rate is (Figure In the scenario where the rate of fixation is the same in and out of the puncta the live-cell is in fixed cells of the punctate percentage However, when one fixation rate is faster than the we observe a bifurcating the fixation rate the puncta is greater than the puncta the fixed cell will have a punctate percentage than the live cell, that fixation enhances the apparent LLPS behaviors. the balance is the fixed cells will have diminished apparent LLPS behaviors than in the live cell. For where the punctate percentage is or due to significantly different binding and the dissociation rates or no significant change to LLPS appearance after fixation In suggests that fixation rates in and out of puncta is to cause a fixation artifact of LLPS systems a

A typical human cell is a crowded soup of thousands of different proteins. One way that the cell organizes this complex mix of contents is by creating separate droplets within the cell, like oil in water. These droplets can form through a process known as liquid-liquid phase separation, or LLPS, where specific proteins gather in high concentrations to carry out their cellular roles. The critical role of LLPS in cellular organization means that it is widely studied by biologists. To detect LLPS, researchers often subject the cells to treatments designed to hold all the proteins in place, creating a snapshot of their natural state. This process, known as fixing, allows scientists to easily label a protein with a fluorescent tag, take pictures of the cells, and look at whether the protein forms droplets in its natural state. This is often easier to do than imaging cells live, but it relies on LLPS being well-preserved upon fixation. To test if this is true, Irgen-Gioro, Yoshida et al. looked at protein droplets in live cells, and then fixed the cells to check whether the appearance of the droplets had changed. The images taken showed that fixation could alter the size and number of droplets depending on the protein being studied. To explain why the effects of fixing change depending on the protein, Irgen-Gioro, Yoshida et al. hypothesized that a faster fixation – relative to how quickly proteins can bind and unbind to their droplets – can better preserve the LLPS droplets. They verified their idea using a microscopy technique in which they imaged single molecules, allowing them to see how different fixation speeds relative to protein binding affected the droplets. The work of Irgen-Gioro, Yoshida et al. identifies an important caveat to using fixation for the study of LLPS in cells. Their findings suggest that researchers should be cautious when interpreting the results of such studies. Given that LLPS in cells is an area of research with a lot of interest, these results could benefit a broad range of biological and medical fields. In the future, Irgen-Gioro, Yoshida et al.’s findings could prompt scientists to develop new fixing methods that better preserve LLPS in cells.

The Enlightenment philosophy, particularly the ideas of Thomas Hobbes and his concepts surrounding the State and Society, serves as a philosophical foundation for numerous subsequent discussions in the fields of social and political sciences. Hobbes’ perspective on human nature and his portrayal of the natural state versus civilization are undeniably among the central tenets of modern thought. He characterizes humanity as the ‘wolf-man’ and underscores the necessity of a social contract-based civilized state to ensure security and safeguard collective interests. Hobbes is primarily concerned with peace, and his political framework, rooted in a modern scientific sense, is designed with the explicit goal of establishing and preserving peace. In the natural state, where humanity exists in an uncivilized condition, each individual is self-interested and disconnected, resulting in a landscape fraught with danger and insecurity. However, a glimmer of hope arises from the fact that human awareness of this predicament serves as the very genesis of contemplation and thought directed towards salvation. The cornerstone of security in the natural state is the equality of all individuals within it. According to Hobbes, society does not represent the spontaneous formation of a fundamental condition for collective life but, rather, the conscious regulation of a potentially volatile state perpetually on the brink of conflict. Through surveys, Hobbes asserts that a majority of people within a society willingly delegate their power and authority to an individual or a group, enabling them to consolidate the various wills into a unified collective will through a voting process. This article aims to explore a fundamental shift in ethics as we transition from classical ethics centered around ‘good’ to modern social ethics founded on ‘security’ and ‘stability.’

Hobbes and Locke differ over the state of nature. In Hobbes the state of nature is a state of generalized insecurity. Each person runs the risk of losing everything, and each person has the right of taking anything—another’s life, possessions—whatever seems a help to his own self-preservation and prosperity. In Locke, as Simmons points out,1 it is not easy to say all at once and briefly what the state of nature is. Many different states of nature are recognized. Not all are violent. Not all discourage productive labour. Not all permit wholesale violations in practice of natural law. Not all exist in the absence of some sort of authority or order. Instead, there are a whole range of states of nature, unified not by a set of inconveniences, but by the fact that people stand outside a legitimate political order. It is often thought that Locke’s theory of the state of nature (and of government) not only differs systematically from Hobbes’s, but that it was arrived at in opposition to Hobbes’s. This seems unlikely, for reasons given long ago by Laslett.2 Locke could not be clearer that his target in the Two Treatises is Filmer. And though Filmer’s theory of political power supports a kind of absolutism, as does Hobbes’s, it is Filmer’s sort of absolutism, patriarchal absolutism, and not Hobbes’s, that is at issue.3 Even if the Two Treatises are not directed at Hobbes, they are comparable to Hobbes’s political writings in this respect: they can be understood as having a message for people who believe they are under tyranny and want to escape it. The message of Hobbes’s political treatises is that, unless it puts people at risk of immediate death, they act unjustly if they try to escape or resist tyranny, since

Review of: Images of anarchy: the rhetoric and science in Hobbes’s state of nature, by Ioannis D. Evrigenis, Cambridge, Cambridge University Press, 2014, 309 pp., £60.00 (hardback), ISBN 978-0-5215-1372-2

Perhaps no single idea is more deeply ingrained in the psyche of liberalism than the political and moral salience of the “individual.” In this respect, John Locke is often regarded as perhaps the preeminent founder of liberalism for to many it is in the account of the state of nature in Locke's Second Treatise that modernity was first introduced to the natural rights-bearing, pre-civil individual who made possible the liberal civic person “constituted by moral sovereignty over one's core beliefs and practices.” Yet the state of nature motif also presents something of a paradox for those trying to understand the individualist premises of Locke's political theory. On the one hand, the state of nature is a decidedly abstract account of the rights of the generic individual in a pre-civil condition, and as such looks like a compendium of purely formal logical deductions from a set of distinct moral propositions. On the other hand, Locke presents this theoretical account as the philosophical basis of individual rights, such as the right to own property and the right to rebel against government, that have enormous practical significance in virtually any conceivable form of political life. It is this combination of theory and practice, abstraction and contextualization, that makes the state of nature concept at once so central to Locke's political teaching and yet so difficult to grasp.

In Anarchy, State, and Utopia, Robert Nozick contends that the question of political philosophy is whether there should be a political state at all. Why not, he asks, have anarchy?1 This is a serious question which, he insists, demands consideration prior to any discussion of the proper structure of a political state. State of nature theory provides, in Nozick's estimation, the most profitable starting point for addressing the question of anarchy. In fact, he perceives a two-fold purpose in state of nature theory, the first justificatory, the second explanatory. In its justificatory role, state of nature theory establishes that if one can demonstrate that a political state (a) would be superior to the best situation of anarchy (i.e., state of nature) 2 which can reasonably be hoped for, (b) would come into being without taking any morally impermissible steps, that is, without violating anyone's natural rights, or (c) would be an improvement over the state of nature, then formation of a political state is justified.3 The explanatory purpose of state of nature theory relates to what Nozick calls a fundamental explanation of (a| realm.4 Such an explanation is valuable because it explains the realm in terms of its most basic, important, and unavoidable features. Hence, he maintains that if one can explain the political realm in terms of the nonpolitical (the state of nature), then one has achieved the most desirable and complete explanation of the political realm possible. The plausibility of Nozick's admittedly libertarian 5 political philosophy thus turns in large part upon the state of nature which undergirds it. In this paper 1 suggest that shortcomings in his formulation of that state of nature undermine his political theory's credibility. In particular, I argue that the state of nature he conceives rests upon two

This collection explores eighteenth-century theories of international market competition that continue to be relevant for the twenty-first century. 'Jealousy of Trade' refers to a particular conjunction between politics and the economy that emerged when success in international trade became a matter of the military and political survival of nations. Today, it would be called 'economic nationalism', and in this book Istvan Hont connects the commercial politics of nationalism and globalization in the eighteenth century to theories of commercial society and Enlightenment ideas of the economic limits of politics. The book begins with an analysis of how the notion of 'commerce' was added to Hobbes' 'state of nature' by Samuel Pufendorf. Hont then considers British neo-Machiavellian political economy after the Glorious Revolution. From there he moves to a novel interpretation of the political economy of the Scottish Enlightenment, particularly of David Hume and Adam Smith, concluding with a conceptual history of nation-state and nationalism in the French Revolution. Jealousy of Trade combines political theory with intellectual history, illuminating the past but also considering the challenges of today.

Chapter 1 shows how Typee (1846) and Omoo (1847) speak back to the Enlightenment debate between Hobbes, Locke, and Rousseau on the state of nature. These books are jointly concerned to reject Rousseau’s “pure” state of nature as a fantasy that does not, and cannot, exist in the world. By contrast, they have little difficulty locating Hobbes’s “corrupted” state of nature, whether in the Typee valley or on board a South Seas whaler. The chapter argues that Melville’s state of nature is simply the state of war prior to, or in the absence of, the state as Leviathan (a Hobbesian term in political theory for the state as a “dominant protective agency” with a “monopoly on violence”).

Causes of Variability W hen we compare the individuals of the same variety or subvariety of our older cultivated plants and animals, one of the first points which strikes us is, that they generally differ more from each other than do the individuals of any one species or variety in a state of nature. And if we reflect on the vast diversity of the plants and animals which have been cultivated, and which have varied during all ages under the most different climates and treatment, we are driven to conclude that this great variability is due to our domestic productions having been raised under conditions of life not so uniform as, and somewhat different from, those to which the parent-species had been exposed under nature. There is, also, some probability in the view propounded by Andrew Knight, that this variability may be partly connected with excess of food. It seems clear that organic beings must be exposed during several generations to new conditions to cause any great amount of variation; and that, when the organisation has once begun to vary, it generally continues varying for many generations. No case is on record of a variable organism ceasing to vary under cultivation. Our oldest cultivated plants, such as wheat, still yield new varieties: our oldest domesticated animals are still capable of rapid improvement or modification.

It is further to be observed that just as the self-assertion necessary to the maintenance of society against the state of nature will destroy that society if it is allowed free operation within, so the self-restraint, the essence of the ethical process, which is no less an essential condition of the existence of every polity may, by excess, become ruinous to it. In a large proportion of cases, crime and pauperism have nothing to do with heredity; but are the consequence partly of circumstances and partly of the possession of qualities which under different conditions of life might have excited esteem and even admiration. The benevolence and openhanded generosity which adorn a rich man may make a pauper of a poor one; the energy and courage to which the successful soldier owes his rise, the cool and daring subtlety to which the great financier owes his fortune may very easily