Alveolar macrophages (AMs) are key mediators of lung function and are potential targets for therapies during respiratory infections. TGFβ is an important regulator of AM differentiation and maintenance, but how TGFβ directly modulates the innate immune responses of AMs remains unclear. This shortcoming prevents effective targeting of AMs to improve lung function in health and disease. Here, we leveraged an optimized ex vivo AM model system, fetal liver-derived alveolar-like macrophages (FLAMs), to dissect the role of TGFβ in AMs. Using transcriptional analysis, we first globally defined how TGFβ regulates gene expression of resting FLAMs. We found that TGFβ maintains the baseline metabolic state of AMs by driving lipid metabolism through oxidative phosphorylation and restricting inflammation. To better understand inflammatory regulation in FLAMs, we next directly tested how TGFβ alters the response to TLR2 agonists. While both TGFβ (+) and TGFβ (-) FLAMs robustly responded to TLR2 agonists, we found an unexpected activation of type I interferon (IFN) responses in TGFβ (+) FLAMs and primary AMs. Surprisingly, mitochondrial antiviral signaling protein and the IFN regulator factors 3 and 7 were required for IFN production by TLR2 agonists and the IFN response was dependent on mitochondrial reactive oxygen species. Together, these data suggest that TGFβ modulates AM metabolic networks and innate immune signaling cascades to control inflammatory pathways in AMs.

This chapter outlines the Sale of Goods Act 1979 (SGA), which is the set of legal rules primarily concerned with the sale of goods. It presents the history of sales law, acknowledging that the modern law of sale is largely found in the SGA. The SGA primarily aims to remove the need to consult prior case law. Thus, decisions of courts in cases prior to the Act should only be examined for guidance in cases where the language of the Act is unclear or ambiguous. The chapter then reflects on the future of sales law, which is considered to be the heart of commercial activity.

Bradgate's Commercial Law provides comprehensive coverage of the principles of commercial law alongside critical discussion. It offers rigorous in-depth analysis and a modern, contextual approach which draws links to the practice of commercial law. This edition includes new chapters on international trade, foreign investment, and the future trajectory of the subject. There is expanded coverage of territorial matters, the law of the sea, and maritime delimitation, with the aim to reflect advances in the area. Also, chapters have been updated to include detailed explanation of topics such as international organisations, international environmental law, the law of treaties, and legal subjects. All chapters have been revised to incorporate new developments and all major judgments in the area.

This chapter provides an overview of the nature and sources of commercial law. It explores the definition of commercial law, referencing how it is notoriously difficult to define. Commercial law draws on principles from a number of different areas of jurisprudence, such as contract law, criminal law, and principles from tort, property, equity, and trusts. The key challenge for commercial law is therefore to strike the right balance between the competing principles regardless of how they overlap. The chapter also considers the impact and significance of consumerism, internationalisation, digitalisation, certainty, and good faith on commercial law. It then discusses the role of equity in commercial law.

This chapter focuses on title conflicts in sale transactions. Despite the technological changes in improving data capture, storage, and analysis that can help provide a greater capacity to record tidal and goods information, it is not the panacea to title conflicts arising from information asymmetry since it also occurs due to conflicting interests. Section 21 of the Sale of Goods Act 1979 (SGA) preserves the basic common law principle (nemo dat quod non habet) that is subjected to exceptions. The provision also contains the principle of estoppel. The chapter acknowledges that the nemo dat principle itself is simple but some of its laws and exceptions are outdated.

This chapter tackles the condition of the goods and statutory implied terms. If the terms of the broken contract between the seller and buyer involve a condition, the buyer may be entitled to reject the goods and terminate the contract. Sections 12 to 15 of the Sale of Goods Act 1979 (SGA) indicate the conditions of implied terms. The chapter explains that implied terms require the goods supplied to conform to the buyer's legitimate expectations, which then may allow the buyer to complain about a breach if their expectations are not met. It also considers how the seller's power to exclude the implied terms, or liability for their breach, is limited by the Unfair Contract Terms Act 1977.

AbstractImmune networks that control antimicrobial and inflammatory mechanisms have overlapping regulation and functions to ensure effective host responses. Genetic interaction studies of immune pathways that compare host responses in single and combined knockout backgrounds are a useful tool to identify new mechanisms of immune control during infection. For disease caused by pulmonaryMycobacterium tuberculosisinfections, which currently lacks an effective vaccine, understanding genetic interactions between protective immune pathways may identify new therapeutic targets or disease-associated genes. Previous studies suggested a direct link between the activation of NLRP3-Caspase1 inflammasome and the NADPH-dependent phagocyte oxidase complex during Mtb infection. Loss of the phagocyte oxidase complex alone resulted in increased activation of Caspase1 and IL1β production during Mtb infection, resulting in failed disease tolerance during the chronic stages of disease. To better understand this interaction, we generated mice lacking bothCybb, a key subunit of the phagocyte oxidase, andCaspase1/11. We found thatex vivoMtb infection ofCybb-/-Caspase1/11-/-macrophages resulted in the expected loss of IL1β secretion but an unexpected change in other inflammatory cytokines and bacterial control. Mtb infectedCybb-/-Caspase1/11-/-mice rapidly progressed to severe TB, succumbing within four weeks to disease characterized by high bacterial burden, increased inflammatory cytokines, and the recruitment of granulocytes that associated with Mtb in the lungs. These results uncover a key genetic interaction between the phagocyte oxidase complex and Caspase1/11 that controls protection against TB and highlight the need for a better understanding of the regulation of fundamental immune networks during Mtb infection.

Immune networks that control antimicrobial and inflammatory mechanisms have overlapping regulation and functions to ensure effective host responses. Genetic interaction studies of immune pathways that compare host responses in single and combined knockout backgrounds are a useful tool to identify new mechanisms of immune control during infection. For disease caused by pulmonary Mycobacterium tuberculosis infections, which currently lacks an effective vaccine, understanding genetic interactions between protective immune pathways may identify new therapeutic targets or disease-associated genes. Previous studies suggested a direct link between the activation of NLRP3-Caspase1 inflammasome and the NADPH-dependent phagocyte oxidase complex during Mtb infection. Loss of the phagocyte oxidase complex alone resulted in increased activation of Caspase1 and IL1β production during Mtb infection, resulting in failed disease tolerance during the chronic stages of disease. To better understand this interaction, we generated mice lacking both Cybb , a key subunit of the phagocyte oxidase, and Caspase1/11 . We found that ex vivo Mtb infection of Cybb -/- Caspase1/11 -/- macrophages resulted in the expected loss of IL1β secretion but an unexpected change in other inflammatory cytokines and bacterial control. Mtb infected Cybb -/- Caspase1/11 -/- mice rapidly progressed to severe TB, succumbing within four weeks to disease characterized by high bacterial burden, increased inflammatory cytokines, and the recruitment of granulocytes that associated with Mtb in the lungs. These results uncover a key genetic interaction between the phagocyte oxidase complex and Caspase1/11 that controls protection against TB and highlight the need for a better understanding of the regulation of fundamental immune networks during Mtb infection.

Using machine learning (ML), we interrogated the function of all human-chimpanzee variants in 2,645 human accelerated regions (HARs), finding 43% of HARs have variants with large opposing effects on chromatin state and 14% on neurodevelopmental enhancer activity. This pattern, consistent with compensatory evolution, was confirmed using massively parallel reporter assays in chimpanzee and human neural progenitor cells. The species-specific enhancer activity of HARs was accurately predicted from the presence and absence of transcription factor footprints in each species. Despite these striking cis effects, activity of a given HAR sequence was nearly identical in human and chimpanzee cells. This suggests that HARs did not evolve to compensate for changes in the trans environment but instead altered their ability to bind factors present in both species. Thus, ML prioritized variants with functional effects on human neurodevelopment and revealed an unexpected reason why HARs may have evolved so rapidly.

Alveolar macrophages (AMs) are tissue-resident cells in the lungs derived from the fetal liver that maintain lung homeostasis and respond to inhaled stimuli. Although the importance of AMs is undisputed, they remain refractory to standard experimental approaches and high-throughput functional genetics, as they are challenging to isolate and rapidly lose AM properties in standard culture. This limitation hinders our understanding of key regulatory mechanisms that control AM maintenance and function. In this study, we describe the development of a new model, fetal liver-derived alveolar-like macrophages (FLAMs), which maintains cellular morphologies, expression profiles, and functional mechanisms similar to murine AMs. FLAMs combine treatment with two key cytokines for AM maintenance, GM-CSF and TGF-β. We leveraged the long-term stability of FLAMs to develop functional genetic tools using CRISPR-Cas9-mediated gene editing. Targeted editing confirmed the role of AM-specific gene Marco and the IL-1 receptor Il1r1 in modulating the AM response to crystalline silica. Furthermore, a genome-wide knockout library using FLAMs identified novel genes required for surface expression of the AM marker Siglec-F, most notably those related to the peroxisome. Taken together, our results suggest that FLAMs are a stable, self-replicating model of AM function that enables previously impossible global genetic approaches to define the underlying mechanisms of AM maintenance and function.

Alveolar macrophages (AMs) are a tissue resident macrophage population within the lung that are critical to maintain lung homeostasis and respond to inhaled antigens. Mechanistic investigations of AM function employing genetic approaches like CRISPR-Cas9 are hindered by the inability to obtain large numbers or maintain the AM-likeness of these cells ex vivo. We recently developed fetal liver-derived alveolar-like macrophages (FLAMs), a model that maintains the expression profile and immune functions of AMs long-term ex vivo. Importantly, FLAMs are genetically tractable using CRISPR-Cas9, enabling previously impossible interrogation of AM functions. Leveraging this innovative tool, we generated a genome-wide knockout FLAM library and are now completing forward genetic screens to dissect the regulation of AM maintenance and function. One such screen identified genes that regulate the surface expression of the AM-specific protein, Siglec-F. Among the numerous AM-associated pathways we identified as positive regulators of Siglec-F, the most robust signature was for genes related to the peroxisome. Thus, we hypothesize that the peroxisome is an essential signaling platform for AM functions. To test this prediction further we are using chemical and genetic approaches to dissect peroxisome functions in AMs. We are examining how peroxisomes modulate gene expression and metabolic flux in AMs and determining how this alters AM responses to stimuli. Together, these results highlight the utility of our novel genetic screening platform in FLAMs to unlock our understanding of AM functional mechanisms like never before. This work was supported by the Rackham Research Endowment Award from Michigan State University.

Mycobacterium tuberculosis colonizes, survives, and grows inside macrophages. In vitro macrophage infection models, using both primary macrophages and cell lines, enable the characterization of the pathogen response to macrophage immune pressure and intracellular environmental cues. We describe methods to propagate and infect primary murine bone marrow-derived macrophages, HoxB8 conditionally immortalized myeloid cells, Max Planck Institute alveolar macrophage-like cells, and J774 and THP-1 macrophage-like cell lines. We also present methods on the characterization of M. tuberculosis intracellular survival and the preparation of infected macrophages for imaging.

ABSTRACTHuman accelerated regions (HARs) are sequences that have evolved at an accelerated rate in the human lineage. Some HARs are developmental enhancers. We used a massively parallel reporter assay (MPRA) to identify HARs with enhancer activity in a mammalian testis cell line. A subset of HARs exhibited differential activity between the human and chimpanzee orthologs, representing candidates for underlying unique human male reproductive biology. We further characterized one of these candidate testis enhancers, 2xHAR.238. CRISPR/Cas9-mediated deletion in a testis cell line and mice revealed that 2xHAR.238 enhances expression ofGli2, encoding a Hedgehog pathway effector, in testis Leydig cells. 4C-seq revealed that 2xHAR.238 contacts theGli2promoter, consistent with enhancer function. In adult male mice, deletion of 2xHAR.238 disrupted mouse male-typical behavior and male interest in female odor. Combined, our work identifies a HAR that promotes the expression ofGli2in Leydig cells and may have contributed to the evolution of human male reproductive biology.

Alveolar macrophages (AMs) are a critical element of the innate immune response to inhaled agents, yet functional and genetic studies of this unique macrophage population are lacking. Current strategies to obtain large quantities of AMs are cumbersome and inefficient. This is due largely to both the high cost of time and resources involved in the extraction of AMs and the inability to effectively culture AMs ex vivo for extended periods of time. While bone marrow derived macrophages (BMDMs) are modeled in numerous immortalized cell lines, AMs currently lack an acceptable model that can be used in vitro. Recently, self-replicating cells derived from the fetal mouse liver, termed “MPI” cells, have been shown to possess AM-like characteristics. Here, we show that early after isolation, these cells are SiglecFhi, Cd11chi, and Cd14low, while also expressing high levels of Pparg, Marco, Itgax, and Car4, akin to AMs. Additionally, like AMs, MPI cells effectively efferocytose dead cell debris and phagocytose silica particles. While these cells lose their “AM-likeness” over time, addition of the cytokine TGF-β dramatically delays this shift away from the AM-like phenotype. Gene expression analysis shows that in contrast to cells treated with TGF-β, untreated MPI cells cease expressing Tgfbr1, the receptor for TGF-β, concurrent with the shift away from the AM-phenotype. Further, these cells are amenable to viral transduction, and we have successfully employed CRISPR/Cas9 targeted genetic editing in MPI cells. These findings further our understanding of MPI cells as an accessible and genetically tractable model for AMs that allow for long-term and large-scale studies that are not possible with AMs isolated ex vivo.

Purpose This paper aims to examine the effect of circular economy's ending of waste on marginal property practices. Design/methodology/approach This paper utilises doctrinal and theoretical legal analysis, along with theoretical perspectives and qualitative empirical evidence drawn from non-legal academic disciplines. Findings The current legalistic conception of waste depends on control and value. The indeterminate status of waste as goods at the margins of consumption attracts attention from legal regimes. This process is evidenced by a commercialised treatment of goods at the margins of consumption, limiting the scope of radical marginal property practices such as freeganism (taking goods abandoned by others, to use such goods). Social implications The circular economy aims to end waste. Restriction, and ultimately elimination, of marginal property practices is necessary for circular economy. Freegans will be limited to acting in a "challenge" role, identifying breaches of commercial commodification processes. Control over the use (including disposal) of goods reduces the spaces available for marginal property practices, which in turn raises problematic normative implications for "normal" consumption practices involving waste. Originality/value This is the first examination of the impact of circular economy on freeganism. It is also the first sustained application of marginal property theory (van der Walt, 2009) in a legal analysis of circular economy and waste.

The legal aspects of the circular economy have until recently been subject to only minimal analysis. This is partly due to the absence of specific legal frameworks; what does exist has been exhortatory and ultimately unenforceable. Moves towards imposing legal obligations are growing, and thus the need for deep and broad legal analysis becomes imperative. This chapter outlines some of the legal issues that will need to be considered in any moves towards a circular economy. There is a focus on the importance of property and ownership, as well as a critical examination of the limited extent of academic work in this area. The need to reduce and remove potential contradictions between different legal regimes is highlighted.

Was there anything especially modernising about the late Victorian changes to English commercial law? What institutions changed or continued? Using the theoretical framework of material civilization developed by Fernand Braudel enables a critical analysis suggesting that English commercial law is neither modern nor especially indicative of national identity. Long standing commercial practices across jurisdictions have clear influence. There is also an explanatory aspect to this theoretical application. The importation of institutional foundations from various jurisdictions over the longue durée meant that doctrinal practices were especially able to allow for significant value transfers to the highest levels of material civilization: financial capitalism. Here the legal institution of commercial law relating to finance is addressed in its international context, alongside another institution, that of commerce itself. Within this analysis there is a more concentrated examination of the Amsterdam experience as being the preeminent commercial world city immediately prior to London's dominance. There is also reference to the role played by judges and commentators in the generation of doctrinal structure in the Victorian period, and the way in which the focus there on property fitted closely with the broader jurisprudential framework and the commercial practices spread over space and time.

Intellectual property (IP) law is everywhere. This wide-ranging collection of essays addresses a fundamental issue with IP law—how can it deal with a world where replication of information is essentially costless, and physical replication may be moving (if not at the same pace) in the same direction with 3D printing technology. The book is divided into three parts focusing on UK, USA and Australian perspectives on 3D printing and IP. The first substantive chapter is an abridged version of Lemley’s ground-breaking article ‘IP in a world without scarcity’.1 Each jurisdictional part provides chapters looking at prime elements of the IP firmament (copyright, design rights, trade marks, and patents) finishing with a more general essay on the impact of 3D printing on law.2 The concluding essay co-authored by Mendis, Lemley and Rimmer provides a useful overview of the preceding chapters, drawing out the points of tension evident throughout the three jurisdictions’ IP regimes. It is clear that whilst there is (to use a loaded term) substantial similarity between the three systems, there are aspects where protection for IP rights (IPR) holders, direct and indirect infringers and ‘good faith’ end users vary between the different jurisdictions. Throughout, a wealth of further literature is referred to. This chapter, read with the book as a whole, provides a valuable foundation for further more detailed scholarship on the issues identified.

Chronic bacterial infections are caused by pathogens that persist within their hosts and avoid clearance by the immune system. Treatment and/or detection of such pathogens is difficult, and the resulting pathologies are often deleterious or fatal. There is an urgent need to develop protective vaccines and host-directed therapies that synergize with antibiotics to prevent pathogen persistence and infection-associated pathologies. However, many persistent pathogens, such as Mycobacterium tuberculosis, actively target the very host pathways activated by vaccination. These immune evasion tactics blunt the effectiveness of immunization strategies and are impeding progress to control these infections throughout the world. Therefore, it is essential that M. tuberculosis immune evasion-related pathogen virulence strategies are considered to maximize the effectiveness of potential new treatments. In this review, we focus on how Mycobacterium tuberculosis infects antigen-presenting cells and evades effective immune clearance by the adaptive response through (i) manipulating antigen presentation, (ii) repressing T cell-activating co-stimulatory molecules, and (iii) inducing ligands that drive T cell exhaustion. In this context, we will examine the challenges that bacterial virulence strategies pose to developing new vaccines. We will then discuss new approaches that will help dissect M. tuberculosis immune evasion mechanisms and devise strategies to bypass them to promote long-term protection and prevent disease progression.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.