Reducing Suffering During Conflict: The Interface Between Buddhism And International Humanitarian Law

This article stems from a project launched by the International Committee of the Red Cross (ICRC) in 2017 to examine the degree to which Buddhism might complement or enhance international humanitarian law (IHL), also known as ‘the law of war’ or ‘the law of armed conflict’. Given that Buddhist teachings discourage violence, scholarship has critiqued Buddhists’ involvement in armed conflict rather than considered how Buddhism might contribute to regulating the conduct of hostilities once war has broken out. Yet the Buddhist aim to reduce suffering is particularly relevant during armed conflict, and the empirical realism of early Buddhist texts shows that early Buddhist communities were very much aware of its grim reality. The article investigates the evidence for this empirical realism before exploring a range of concepts, doctrines and practices from within Buddhism that are pertinent to the recognition and implementation of IHL principles and the conduct of war. While IHL lays down explicit rules to follow during war, Buddhism emphasises broader ethical principles to be applied, so as not to dilute its ideal of non-violence. At a deeper level, it addresses the intention or motivation of parties to armed conflict, and possesses psychological insights and resources to help change their behaviour

DNA compaction induced by a cationic polymer or surfactant impact gene expression and DNA degradation

There is an increasing interest in achieving gene regulation in biotechnological and biomedical applications by using synthetic DNA-binding agents. Most studies have so far focused on synthetic sequence-specific DNA-binding agents. Such approaches are relatively complicated and cost intensive and their level of sophistication is not always required, in particular for biotechnological application. Our study is inspired by in vivo data that suggest that DNA compaction might contribute to gene regulation. This study exploits the potential of using synthetic DNA compacting agents that are not sequence-specific to achieve gene regulation for in vitro systems. The semi-synthetic in vitro system we use include common cationic DNA-compacting agents, poly(amido amine) (PAMAM) dendrimers and the surfactant hexadecyltrimethylammonium bromide (CTAB), which we apply to linearized plasmid DNA encoding for the luciferase reporter gene. We show that complexing the DNA with either of the cationic agents leads to gene expression inhibition in a manner that depends on the extent of compaction. This is demonstrated by using a coupled in vitro transcription-translation system. We show that compaction can also protect DNA against degradation in a dose-dependent manner. Furthermore, our study shows that these effects are reversible and DNA can be released from the complexes. Release of DNA leads to restoration of gene expression and makes the DNA susceptible to degradation by Dnase. A highly charged polyelectrolyte, heparin, is needed to release DNA from dendrimers, while DNA complexed with CTAB dissociates with the non-ionic surfactant C12E5. Our results demonstrate the relation between DNA compaction by non-specific DNA-binding agents and gene expression and gene regulation can be achieved in vitro systems in a reliable dose-dependent and reversible manner

Proxy agents, auxiliary forces, and sovereign defection: assessing the outcomes of using non-state actors in civil conflicts

This article interrogates the role of non-state armed actors in the Ukrainian civil conflict. The aim of this article is twofold. First, it seeks to identify the differences between the patterns of military intervention in Crimea (direct, covert intervention), and those in the South-East (mixed direct and indirect – proxy – intervention). It does so by assessing the extent of Russian troop involvement and that of external sponsorship to non-state actors. Second, it puts forward a tentative theoretical framework that allows distinguishing between the different outcomes the two patterns of intervention generate. Here, the focus is on the role of non-state actors in the two interventionist scenarios. The core argument is that the use of nonstate actors is aimed at sovereign defection. The article introduces the concept of sovereign defection and defines it as a break-away from an existing state. To capture the differences between the outcomes of the interventions in Crimea and South-East, sovereign defection is classified into two categories: inward and outward. Outward sovereign defection is equated to the territorial seizure of the Crimean Peninsula by Russian Special Forces, aided by existing criminal gangs acting in an auxiliary capacity. Inward sovereign defection refers to the external sponsorship of the secessionist rebels in South-East Ukraine and their use as proxy forces with the purpose of creating a political buffer-zone in the shape of a frozen conflict. To demonstrate these claims, the article analyses the configuration of the dynamics of violence in both regions. It effectively argues that, in pursuing sovereign defection, the auxiliary and proxy forces operate under two competing dynamics of violence, delegative and non-delegative, with distinct implications to the course and future of the conflict

DNA condensation using G4 dendrimers and CTAB surfactants.

<p>(A) The fluorescence intensity of GelStar bound to DNA, shown as a function of <i>r</i>_charge in solutions containing 10 mM NaBr for G4 dendrimers (▵) and CTAB (•). Data are normalized to the amounts produced in the samples only containing DNA (in the absence of dendrimer or surfactant) and the <i>I</i>_max value is linearly dependent on the amount of DNA that is available to bind GelStar. The DNA concentration is 2 μg mL⁻¹ and error bars are smaller or equal to the size of the markers. (B) The mean number of G4 per compacted DNA chain at varying <i>r</i>_charge, calculated as described in the text. Note that below charge neutralization the amount of bound G4 per DNA strand is constant, that is each complex contains the same number of G4. Once the neutralization point is reached, the solution only contains compacted DNA and the number of G4 per compacted DNA increases. The results from the electrophoreses study - DNA condensation by G4 dendrimers and CTAB surfactants - are shown in (C) (D), respectively. Lane 1 in both C and D displays free linearized plasmid DNA in the absence of any compacting agent (control, 4331 bp). Samples in lanes 2–11 contain increasing amounts of the compacting agent and the corresponding <i>r</i>_charge values are indicated. The DNA concentration was 25 μg mL⁻¹ and the gels were stained with Ethidium Bromide (EtBr).</p

Cryo-TEM images of DNA (0.1 mg mL⁻¹) complexes.

<p>(A, C) G4/DNA of <i>r</i>_charge  = 0.5, and (B, D) CTAB/DNA of <i>r</i>_charge  = 7.5. All samples were prepared in aqueous solutions of 10 mM NaBr, but (A) and (B) display G4/DNA and CTAB/DNA complexes, respectively, after being transferred to the tRB used for <i>in vitro</i> transcription/translation experiments. (C) and (D) are the reference samples in the 10 mM NaBr solution used for DNA compaction. Scale bars are 100 nm, the arrow indicates frost (artifact), the white stars indicate the carbon film and the black star shows a fracture of the vitreous film.</p

Luciferase gene expression and DNA accessibility as a function of <i>r</i>_charge using pre-casted RNA gels.

<p>(A) G4 dendrimers and (B) CTAB. The synthesized amounts of RNA are displayed and samples were not pretreated with Dnase I. References are displayed in B where lane 1 shows the sample consisting only of DNA and without any compacting agent or transcriptional activity. Lane 2 shows the control sample containing DNA and the <i>in vitro</i> transcription mixture in the absence of compacting agents. Gels were post-stained using GelStar.</p

Protection of DNA against Dnase I digestion using gels stained with GelStar.

<p>(A) G4/DNA complexes with <i>r</i>_charge  = 0.4 are used and the untreated complex is loaded on lane 1. The 2^nd lane displays the dissociated complex after treatment with 10 μg mL⁻¹ heparin for 30 min. All other samples (lanes 3–7) are treated with 1 unit of Dnase I for the indicated time periods. To the samples in lanes 4–7, heparin was added after the Dnase I enzyme was heat inactivated. (B) Linearized plasmid DNA only is loaded onto lane 1 and the sample loaded onto lane 2 contains DNA, treated with Dnase I for 30 min. Samples loaded onto lanes 3–7 contain DNA and CTAB (<i>r</i>_charge  = 7.5). Samples loaded onto lanes 4–7 are treated with Dnase I for the time periods indicated.</p

Cryo-TEM of DNA dissolved in the tRB used for <i>in vitro</i> transcription and translation experiments.

<p>Coexisting domains of free DNA (A) and tightly packed clusters of DNA (B) are shown. Scale bars are 100 nm and the DNA concentration is 0.1 mg mL⁻¹.</p

The solvent effect on DNA compaction using steady state fluorescence spectroscopy.

<p>Columns marked as NaBr correspond to samples in aqueous solutions of 10<i>in vitro</i> gene expression kit. Data are normalized to the emitted intensity in the sample only containing DNA (without compacting agent) in 10 mM NaBr solution (1^st column). The <i>r</i>_charge values reported for the samples containing compacted DNA are 7.5 for CTAB/DNA and 0.5 for G4/DNA. The DNA concentration is 2 μg mL⁻¹ and the dye used is GelStar.</p

DNA condensation using G4 dendrimers and CTAB surfactants.

<p>(A) The fluorescence intensity of GelStar bound to DNA, shown as a function of <i>r</i>_charge in solutions containing 10 mM NaBr for G4 dendrimers (▵) and CTAB (•). Data are normalized to the amounts produced in the samples only containing DNA (in the absence of dendrimer or surfactant) and the <i>I</i>_max value is linearly dependent on the amount of DNA that is available to bind GelStar. The DNA concentration is 2 μg mL⁻¹ and error bars are smaller or equal to the size of the markers. (B) The mean number of G4 per compacted DNA chain at varying <i>r</i>_charge, calculated as described in the text. Note that below charge neutralization the amount of bound G4 per DNA strand is constant, that is each complex contains the same number of G4. Once the neutralization point is reached, the solution only contains compacted DNA and the number of G4 per compacted DNA increases. The results from the electrophoreses study - DNA condensation by G4 dendrimers and CTAB surfactants - are shown in (C) (D), respectively. Lane 1 in both C and D displays free linearized plasmid DNA in the absence of any compacting agent (control, 4331 bp). Samples in lanes 2–11 contain increasing amounts of the compacting agent and the corresponding <i>r</i>_charge values are indicated. The DNA concentration was 25 μg mL⁻¹ and the gels were stained with Ethidium Bromide (EtBr).</p