bluuebunny/crossref_metadata_2025 huggingface dataset:
Rhizome
Authors: Colebrook Claire
Published: January 2021

The concept of the rhizome was first articulated in Gilles Deleuze and Felix Guattari’s Kafka: Toward a Minor Literature, published in French in 1975 and translated into English in 1986. Here the term emerges from a reading of Kafka’s description of movements in his novels and short stories, but it is also tied to a mode of reading and of composition. In Mille Plateaux (1980), translated into English as A Thousand Plateaus in 1987, the term has both a broad reference towards modes of thinking and analyzing that are nonhierarchical and decentered, and a more specifically literary sense of styles of writing. In A Thousand Plateaus, the term is introduced in order to describe a mode of composition that is distinct from the book, and a theory of language that is opposed to a basic structure, logic, or grammar from which variations develop. Languages and dialects do not emerge from a central grammar; instead, everything begins with variations of sound and sense. There is no universal grammar; every language has its distinct mode of growth. It is therefore illegitimate to talk of grammar “trees,” and far better to think of variation without a center. Rather than a linear development or progression, a rhizomatic text is composed of multiple points of entry. A rhizome is a lateral, decentered, proliferating, and interconnected web of relations and is therefore unlike the hierarchical (root, branch, offshoots) model of a tree.
Digital Humanities
Authors: Burrows Simon , Falk Michael
Published: January 2021

The article offers a definition, overview, and assessment of the current state of digital humanities, particularly with regard to its actual and potential contribution to literary studies. It outlines the history of humanities computing and digital humanities, its evolution as a discipline, including its institutional development and outstanding challenges it faces. It also considers some of the most cogent critiques digital humanities has faced, particularly from North American-based literary scholars, some of whom have suggested it represents a threat to centuries-old traditions of humanistic inquiry and particularly to literary scholarship based on the tradition of close reading. The article shows instead that digital humanities approaches gainfully employed offer powerful new means of illuminating both context and content of texts, to assist with both close and distant readings, offering a supplement rather than a replacement for traditional means of literary inquiry. The digital techniques it discusses include stylometry, topic modeling, literary mapping, historical bibliometrics, corpus linguistic techniques, and sequence alignment, as well as some of the contributions that they have made. Further, the article explains how many key aspirations of digital humanities scholarship, including interoperability and linked open data, have yet to be realized, and it considers some of the projects that are currently making this possible and the challenges that they face. The article concludes on a slightly cautionary note: What are the implications of the digital humanities for literary study? It is too early to tell.
Attitudes Toward LGB Families: International Policies and LGB Family Planning
Authors: Costa Pedro Alexandre
Published: January 2021

According to recent U.S. census data, there are over 700,000 same-gender couples, of which 114,00 have children. U.K. census data further revealed over 200,000 same-gender parented families, and there is evidence that these numbers have been increasing in the last few decades. Between the late 1980s and early 1990s, research on the psychosocial well-being of LGB families was established with a focus on the potential impact of parents’ sexual orientation on the psychological adjustment of their children. Interest in LGB families was evidenced by the growing political and public attention, and became a central issue within the LGBT+ movement across the Western world, especially in Europe and the United States. However, attitudes toward LGB family policies have not evolved in a linear fashion insofar as they have accompanied the constant back and forth in LGB family policies and legislation. Negative attitudes toward LGB family policies are rooted in the negative evaluations of LGB individuals based on beliefs that LGB people are less fit as parents or unable to form and sustain healthy relationships because of their sexual or gender identity. However, these negative beliefs differ according to heterosexual individuals’ characteristics. Research has shown that men, older, less educated, non-White, politically conservative, highly religious, and authoritarian, as well as those who believe that homosexuality is controllable, strictly adhere to traditional gender roles and authorities, and do not have frequent or close contact with LGB individuals, hold higher levels of sexual prejudice toward LGB individuals and LGB family policies.

As of January 2020, same-gender marriage and parenthood are recognized in around 30 countries worldwide, although some countries recognize some forms of same-gender unions, but not marriage, whereas others recognize the right of LGB individuals to have children but not to marry. LGB family policies have progressed mostly through two different pathways: (a) the judicial pathway, which has involved litigation and court rulings on specific matters related to same-gender relationships and parenthood and which was undertaken in the United States, and (b) the legislative pathway, which has relied on political discussion and policy initiatives and was undertaken in the Iberian Peninsula (Portugal and Spain). The different pathways to equality in LGB family policies have different impacts for LGB individuals. In particular, the constant negative messages regarding same-gender couples as being unable to have healthy relationships have been shown to contribute to chronic minority stress and psychological distress among LGB individuals. By contrast, the legalization of same-gender marriage and parenthood provide important benefits and protections for LGB families in addition to promoting their well-being. Examining the evolution of attitudes and legislation regarding LGB family policies is important to inform further initiatives aimed at correcting inequalities for LGB families.
Queer as Materialism
Authors: Noyé Sophie , Rebucini Gianfranco
Published: January 2021

Since the 2000s, forms of articulation between materialist and Marxist theory and queer theory have been emerging and have thus created a “queer materialism.” After a predominance of poststructuralist analyses in the social sciences in the1980s and 1990s, since the late 1990s, and even more so after the economic crisis of 2008, a materialist shift seems to be taking place. These recompositions of the Marxist, queer, and feminist, which took place in activist and academic arenas, are decisive in understanding how the new approaches are developing in their own fields.

The growing legitimacy of feminist and queer perspectives within the Marxist left is part of an evolution of Marxism on these issues. On the other side, queer activists and academics have highlighted the economic and social inequalities that the policies of austerity and capitalism in general induce among LGBTQI people and have turned to more materialist references, especially Marxist ones, to deploy an anticapitalist and antiracist argument. Even if nowadays one cannot speak of a “queer materialist” current as such, because the approaches grouped under this term are very different, it seems appropriate to look for a “family resemblance” and to group them together. Two specific kinds of “queer materialisms” can thus be identified. The first, queer Marxism, seeks to theorize together Marxist and queer theories, particularly in normalization and capitalist accumulation regimes. The second, materialist queer feminism, confronts materialist/Marxist feminist thought with queer approaches and thus works in particular on the question of heteropatriarchy based on this double tradition.
The Politics of Same-Sex Marriage Reforms
Authors: Isailović Ivana
Published: January 2021

Over the past few years, same-sex marriage reforms have become central to contemporary LGBTQ movements. As a result of their mobilizations, many countries across the world have adopted same-sex marriage reforms. According to scholars, LGBTQ movements were successful in part because they used law and legal discourse, arguing that same-sex marriage flows from states’ legal obligations to protect equality and prohibit discrimination.

The turn to law and the law of marriage in the local and transnational contexts may fail, however, to deliver substantive justice for all LGBTQ people. First, same-sex marriage reforms, rather than being just a translation of equality into law, is a product of ideological and legal battles in specific socioeconomic contexts. For instance, in the United States, same-sex marriage, rather than being another form of relationship recognition, became prominent because of the centrality of marriage in the country’s economic, cultural, and legal order. Second, the law of marriage is a system of governance historically shaped by different-sex couples’ needs, with specific one-size-fits-all rules that may not correspond to LGBTQ individuals’ desires, wishes, and lived experiences. Third, as queer theorists have shown, the law of marriage creates an “outside,” a space of exclusion that is inseparable from the legal regime of marriage and the cultural intelligibility of marriage. The emphasis on marriage by LGBTQ movements risks delegitimizing other forms of intimate relationships. The emphasis on marriage may also entrench neoliberalism in contexts in which the marriage, not the state, is seen as a primary safety net. Finally, in the global or transnational setting, claims for same-sex marriage may perhaps unintentionally feed into representations of civilizational conflicts, between those countries that recognize same-sex marriage and those that do not, while also erasing the variety of local practices around sexuality and gender norms.
Vulnerable Groups During Crisis
Authors: DeYoung Sarah E.
Published: January 2021

During crisis events such as humanitarian conflicts, population displacement, natural disasters, and others, some people are more vulnerable to long-term physical, psychological, and overall adverse outcomes. Aspects of context that affect vulnerability include: (a) the nature of the hazard or conflict event; (b) the geographic location and structural surroundings; and (c) involvement of key groups during crisis. The nature of the event includes barriers for access to well-being in high-income and low-income contexts, the speed of onset of the hazard, the scope and type of hazard (localized or catastrophic, natural or technological, and other factors). Geographic location and structural surroundings include factors such as isolation caused by an island context, structural mitigation (such as earthquake-resistant construction), pollution and environmental exposure, and implementation of land use planning or sustainable farming. Finally, with regard to involvement of key groups in crisis, it is important to consider ways in which group coordination, logistics, cultural competency, public policy, social movements, and other mechanisms can exacerbate or improve conditions for vulnerable groups.

Groups more likely to experience adverse outcomes in disasters include: ethnic and racial minoritized persons, people considered to be low caste, women, children, infants, sexual minorities, religious minorities, elders, and immigrants and refugees. Displacement and relocation are associated with specific increases in exposure to food insecurity, human trafficking, and reduced access to reproductive care. Contextual factors are also related to the severity of the adverse outcomes these groups experience in crisis. These factors include access to healthcare, access to education, and economic status. There are also unique groups whose needs, social systems, and cultural factors increase barriers to evacuation, accessing warning information, or accessing safe sheltering. These groups include persons with functional and access needs including medical or cognitive impairments, elderly individuals, people with companion animals, and people with mental illness.
The EU Migration Crisis: A Crisis Analysis Case Study
Authors: Slominski Peter
Published: January 2021

The European Union (EU) migration crisis has been part and parcel of a conglomerate of crises that have affected the EU since the late 2000s, as have the financial and sovereign debt crisis, “Brexit,” the Russia–Ukraine conflict, as well as tensions within transatlantic relations. Scholarship on the EU has devoted much attention in assessing what the migration crisis means for EU integration at large. In particular, EU scholars are interested why the migration crisis has led to political gridlock and a renationalization of border controls rather than a deepening of integration. While they differ in their explanations, these explanations shed light on different aspects of the crisis and are far from mutually exclusive.

Scholars who are more interested in the area of EU Justice and Home Affairs (JHA) largely agree with EU theorists that the field suffers from an incomplete governance design, the dominance of EU member states, and weak supranational capacities. Their analysis also focuses on intra-EU dynamics but offers a more nuanced empirical assessment of relevant EU institutions and decision-making in the course of managing the migration crisis. This growing body of research produces valuable insights and largely confirms existing scholarship, including that on the growing securitization and externalization of EU asylum and migration policy. The EU’s understanding as a norm-based power is particularly challenged by the migratory movements in the wake of the crisis. A small but growing scholarship analyses how the EU is balancing its non-entrée policy with its legal obligation, and what kind of governance arrangements result from that.

While this scholarship has enriched our understanding of the EU migration crisis, it has not generated a major refinement of the standard approaches of EU theorists and JHA scholars. To further enrich the literature on the migration crisis, scholars should go beyond studying the dynamics of EU decision-making and the role of EU institutions. Such an approach should engage more systematically with international actors and institutions that have the capacity to influence EU migration policy. At the same time, global phenomena such as war, poverty, or climate change should also be taken into account in assessing the EU’s room for maneuver in handling migratory pressures. Future research on the migration crisis as well as on migration challenges should thus not only connect with other subfields of political science, such as policy analysis or international relations, but also open up to other disciplines such as law, demography, or environmental studies.
Canada: Very “Civil” Military Relations
Authors: Sokolsky Joel J.
Published: January 2021

Civil–military relations in Canada are “civil” not because Canadians are inherently “nice,” but because there is not much opportunity, incentive, or fodder for serious and intensive disagreement over defense policy between the military and the political leadership. This does not mean that disagreements do not arise or that the military has not chaffed from time to time under Canada’s relatively strict liberal-democratic traditions of civilian control over the military.

Moreover, it is not the structure, traditions, and practices of government in Canada that best explain the relatively ordered nature of civil–military relations. Rather, one must look to the very nature of the Canadian defense and security situation. While Canada is also known for, and at times has vigorously promoted, its contributions to collective security through United Nations (UN) peacekeeping operations, it has been its participation in the North Atlantic Treaty Organization (NATO) and the North American Aerospace Defence Command (NORAD) that has determined almost all of defense policy and the posture and major weapons systems of the Canadian Armed Forces (CAF). This has had profound implications for civil–military relations in Canada and also accounts for the relatively “civil” tone, and the lack of tension and controversy, in those relations. This has meant that Canada’s political leaders and the senior leadership of the CAF have not had to engage in intensive “dialogue” on alternative conceptions of the international strategic environment and what a Canadian “grand strategy” should be in order to guide how the Canadian forces should be postured and equipped to meet security challenges. Canada does have a “grand strategy”; it is just not that grand.

In the end, what tempers and smooths civil–military relations in Canada is that the CAF remains supportive of Canada’s not-so-grand grand strategy and accompanying expeditionary strategic culture, even if that culture is disciplined by a fiscal efficiency that sometimes seeks to extract as much international involvement out of as few military assets as possible. It is well understood on both sides of the civil–military divide that the CAF needs to maintain the confidence of political leaders to be effective in supporting the foreign policy objectives of the government through the applied or apprehended use of force when requested. In order to achieve this, the senior officer corps and the military apparatus must be cognizant of the breadth of nonmilitary factors that play into civil–military relations. Acknowledging that the scope of policy choices is inherently limited by international factors beyond Canada’s control, the importance of domestic priorities, and the access that Canadian politicians have to military expertise and assessments from other sources, the CAF has, for the most part, unobtrusively provided its political master with advice consistent with the realities facing Canadians at home and abroad. Even here, however, the Canadian military, perhaps more than most, understands the unequal character of its dialogue.
Protected Environmental Areas across Brazil
Authors: Freitas Frederico
Published: January 2021

Brazil, the fifth-largest country on earth, has almost a third of its territory classified as protected areas. They include nature reserves with the strictest level of protection and areas reserved for sustainable use. They comprise, in their majority, public lands, but contain a percentage of privately owned estates. They are regulated by a centralized national system of protected areas, but their management is fragmented at all levels of government—federal, state, and municipal. They are located in the sparsely inhabited Brazilian hinterland but can also be found close to the country’s large and densely populated cities. And throughout the years, they have appealed to a diversity of values and policy goals to justify their existence: protection of natural features, development of frontiers, preservation of endangered species and their inhabitants, conservation of biodiversity, sustainability, social justice. All in all, the protected areas of Brazil represent one of the most extensive and ambitious forms of territorial intervention ever implemented in the country.

It was not always like that. With a few exceptions, protected areas controlled by the state for the sake of conserving natural landscapes and resources were mostly absent before the 20th century. It was only in the 1930s that the Vargas regime established the first national parks in Brazil. They were followed by a dozen other parks in the 1950s and 1960s, but up until the 1970s, the establishment of protected areas in Brazil lacked proper planning, institutional support, and policy goals. This situation changed in the mid-1970s, during the height of the military dictatorship, when an alliance of Brazilian and foreign conservationists began pushing for the inclusion of protected areas in Amazonian development programs. The adoption of the language and methods of dictatorship-era territorial planning by the proponents of conservation set the basis for the development of a national system of protected areas. The system took twenty years to be passed into law, a period that coincided with the end of twenty-one years of military dictatorship and the return to democracy in Brazil. One of the novelties of this period was the appearance of groups, such as Amazonian rubber tappers, with a stake in defining conservation policy. They introduced ideas of social justice in the conservation debate, in what came to be known as socio-environmentalism. It was also during the post-dictatorship period that Brazilian conservationism adopted global standards of conservation, for example protection of biodiversity and sustainable development, as justifiable policy goals. In the early 21st century Brazil counts over 2,300 distinct protected areas distributed throughout all of its twenty-six states. They are part of a single, but diverse, national system of protected areas.
Digital Resources: Study of Danish Activities in the Caribbean
Authors: Simonsen Gunvor
Published: January 2021

From the 1670s to 1917, Denmark (until 1814 Denmark–Norway) maintained colonies in the eastern Caribbean. The island of St. Thomas was colonized in 1672, St. John in 1718, and St. Croix was bought from the French in 1733. Racial slavery soon came to dominate the Danish islands and was only abolished in 1848. Most people arrived to the islands as captive Africans, while most Europeans were of either Dutch or British origin. In 1917, the islands, constituting the Danish West Indies, were sold to the United States of America and became the US Virgin Islands. As part of the centennial of 2017, commemorating the transfer of the Virgin Islands to the United States of America, major Danish cultural institutions, such as the National Archives, the Royal Library, and the National Museum, digitized large collections concerning Danish activities and Danish rule in the Caribbean, including the archive of the Danish West India and Guinea Company, the archives of local government agencies in the Caribbean, large collections of photos, drawings, and maps, as well as a significant part of the written works concerning the Danish West Indies published prior to 1917. In combination with older digital platforms, new online resources facilitate the triangulation of many different kinds of evidence, which in turn promises to generate fascinating new histories of the people who lived in the US Virgin Islands while they were under Danish rule.
Comparative Ultrastructural Analysis of Thalamocortical Innervation of the Primary Motor Cortex and Supplementary Motor Area in Control and MPTP-Treated Parkinsonian Monkeys
Authors: Villalba Rosa M , Behnke Joseph A , Pare Jean-Francois , Smith Yoland
Published: January 2021

The synaptic organization of thalamic inputs to motor cortices remains poorly understood in primates. Thus, we compared the regional and synaptic connections of vGluT2-positive thalamocortical glutamatergic terminals in the supplementary motor area (SMA) and the primary motor cortex (M1) between control and MPTP-treated parkinsonian monkeys. In controls, vGluT2-containing fibers and terminal-like profiles invaded layer II–III and Vb of M1 and SMA. A significant reduction of vGluT2 labeling was found in layer Vb, but not in layer II–III, of parkinsonian animals, suggesting a potential thalamic denervation of deep cortical layers in parkinsonism. There was a significant difference in the pattern of synaptic connectivity in layers II–III, but not in layer Vb, between M1 and SMA of control monkeys. However, this difference was abolished in parkinsonian animals. No major difference was found in the proportion of perforated versus macular post-synaptic densities at thalamocortical synapses between control and parkinsonian monkeys in both cortical regions, except for a slight increase in the prevalence of perforated axo-dendritic synapses in the SMA of parkinsonian monkeys. Our findings suggest that disruption of the thalamic innervation of M1 and SMA may underlie pathophysiological changes of the motor thalamocortical loop in the state of parkinsonism.
Acute pulmonary embolism in a patient with mild COVID-19 symptoms: a case report
Authors: Fiorini Norberto B , Garagoli Fernando , Bustamante Rosana C , Pizarro Rodolfo
Published: January 2021

Background

The venous thromboembolism (VTE) is a frequent condition, which may worsen the prognosis of hospitalized COVID-19 patients. Nevertheless, the incidence of this complication is unknown in patients with mild COVID-19 symptoms.

Case summary

A 26-year-old female nurse, who had been taking oral contraceptive pills (OCPs) treatment for the last 2 years, developed mild COVID-19 symptoms (rhinitis and anosmia). She underwent isolation at home and was subsequently followed up with telehealth visits. Fifteen days after her initial presentation, she developed acute onset sudden dyspnoea. On physical examination, she was found to be tachycardic with normal pulse oximetry. The initial risk score for VTE was moderate and laboratory results showed increased D-dimer level without other relevant findings. Computed tomography pulmonary angiography was performed, which confirmed low-risk subsegmental pulmonary embolism.

Discussion

Venous thromboembolism in patients who present with severe COVID-19 symptoms has already been described in the literature; its incidence is greater in patients hospitalized in intensive care units. Efforts to prevent VTE based on risk scores are widely recognized. However, the relationship in patients who present with mild COVID-19 symptoms and VTE is still unknown. Recently, experts on this field have introduced thromboprophylaxis guidelines including ambulatory patients based on the severity of COVID-19 symptoms and pro-thrombotic risk. Our patient showed no major risk for developing VTE; therefore, the VTE could be associated with SARS-CoV-2 infection or the eventual pro-thrombotic association with the concomitant use of OCPs.
Subannular repair for functional mitral regurgitation type IIIb in patients with ischaemic versus dilated cardiomyopathy
Authors: Pausch Jonas , Sequeira Gross Tatiana , Müller Lisa , von Stumm Maria , Kloth Benjamin , Reichenspurner Hermann , Girdauskas Evaldas
Published: January 2021

OBJECTIVES

Functional mitral regurgitation (FMR) is a sequel of left ventricular (LV) remodelling in heart failure patients. Relocation of both papillary muscles aims to specifically address mitral leaflet tethering to improve long-term durability of modern FMR repair. Nevertheless, the prognostic impact of the underlying cardiomyopathy on the outcome after FMR repair is unknown.

METHODS

We analysed 84 consecutive heart failure patients with severe FMR, LV ejection fraction <40%, LV end-diastolic diameter ≥55 mm and tenting height >10 mm, who underwent ring annuloplasty and simultaneous bilateral papillary muscles relocation between June 2016 and March 2019. One-year outcome of 54 patients with ischaemic cardiomyopathy (‘ICM group’) was prospectively compared to the remaining 30 patients with dilated cardiomyopathy (‘DCM group’).

RESULTS

One-year survival was similar in both groups (96% in the ‘ICM group’ vs 97% in the ‘DCM group’; P = 0.93). Furthermore, primary composite outcome (i.e. freedom from death or mitral regurgitation ≥ 2) at 1-year postoperatively was comparable between the study groups (94%in the ‘ICM group’ vs 87% in the ‘DCM group’; P = 0.10). LV end-diastolic diameter 1-year after surgery was significantly reduced, as compared to preoperative values, in the ‘DCM group’ (P = 0.018), but not in the ‘ICM group’ (P = 0.058). Improvement of New York Heart Association functional class and reduction of serum levels of N-terminal pro-B natriuretic peptide at 1 year was comparable in both study groups.

CONCLUSIONS

Standardized relocation of both papillary muscles to correct FMR resulted in very satisfactory in-hospital and 1-year outcomes, in both ICM and DCM. DCM patients showed similar improvement in heart failure symptoms and LV re-remodelling compared to ICM patients. Subannular repair is developing towards a valid therapeutic option in heart failure patients presenting with severe FMR.
Eif2b3 mutants recapitulate phenotypes of vanishing white matter disease and validate novel disease alleles in zebrafish
Authors: Lee Yu-Ri , Kim Se Hee , Ben-Mahmoud Afif , Kim Oc-Hee , Choi Tae-Ik , Lee Kang-Han , Ku Bonsu , Eum Juneyong , Kee Yun , Lee Sangkyu , Cha Jihoon , Won DongJu , Lee Seung-Tae , Choi Jong Rak , Lee Joon Soo , Kim Heung Dong , Kim Hyung-Goo , Bonkowsky Joshua L , Kang Hoon-Chul , Kim Cheol-Hee
Published: January 2021

Leukodystrophy with vanishing white matter (VWM), also called Childhood Ataxia with Central Nervous System Hypomyelination, is caused by mutations in the subunits of the eukaryotic translation initiation factor, EIF2B1, EIF2B2, EIF2B3, EIF2B4 or EIF2B5. However, little is known regarding the underlying pathogenetic mechanisms, and there is no curative treatment for VWM. In this study, we established the first EIF2B3 animal model for VWM disease in vertebrates by CRISPR mutagenesis of the highly conserved zebrafish ortholog eif2b3. Using CRISPR, we generated two mutant alleles in zebrafish eif2b3, 10- and 16-bp deletions, respectively. The eif2b3 mutants showed defects in myelin development and glial cell differentiation, and increased expression of genes in the induced stress response pathway. Interestingly, we also found ectopic angiogenesis and increased VEGF expression. Ectopic angiogenesis in the eif2b3 mutants was reduced by the administration of VEGF receptor inhibitor SU5416. Using the eif2b3 mutant zebrafish model together with in silico protein modeling analysis, we demonstrated the pathogenicity of 18 reported mutations in EIF2B3, as well as of a novel variant identified in a 19-month-old female patient: c.503 T > C (p.Leu168Pro). In summary, our zebrafish mutant model of eif2b3 provides novel insights into VWM pathogenesis and offers rapid functional analysis of human EIF2B3 gene variants.
Collective Deterrence in the Shadow of Shifting Power
Authors: Phillips Julianne , Wolford Scott
Published: January 2021

Twelve of twenty-six war-winning coalitions since 1815 have seen at least two members go to war against one another after victory. What separates durable and fragile war-winning coalitions? To answer this question, we analyze a game-theoretic model of shifting intra-coalition power and collective deterrence. We show that (1) shifting power within war-winning coalitions can undermine commitments to the postwar settlement, but (2) revisionist threats from a powerful defeated side can enhance the credibility of commitments within the winning coalition, securing peace when intra-coalition war would otherwise be inevitable. We also recover these patterns in empirical models of the outbreak of war between former coalition partners: shifting power within a coalition is associated with increased probabilities of intra-coalition war, but only when the defeated side is not too powerful. A common enemy can thus preserve peace between former partners who would otherwise go to war over the terms of shared victory.
Enhanced production of clavulanic acid by improving glycerol utilization using reporter-guided mutagenesis of an industrial Streptomyces clavuligerus strain
Authors: Shin Chang-Hun , Cho Hang Su , Won Hyung-Jin , Kwon Ho Jeong , Kim Chan-Wha , Yoon Yeo Joon
Published: January 2021

Clavulanic acid (CA) produced by Streptomyces clavuligerus is a clinically important β-lactamase inhibitor. It is known that glycerol utilization can significantly improve cell growth and CA production of S. clavuligerus. We found that the industrial CA-producing S. clavuligerus strain OR generated by random mutagenesis consumes less glycerol than the wild-type strain; we then developed a mutant strain in which the glycerol utilization operon is overexpressed, as compared to the parent OR strain, through iterative random mutagenesis and reporter-guided selection. The CA production of the resulting S. clavuligerus ORUN strain was increased by approximately 31.3% (5.21 ± 0.26 g/l) in a flask culture and 17.4% (6.11 ± 0.36 g/l) in a fermenter culture, as compared to that of the starting OR strain. These results confirmed the important role of glycerol utilization in CA production and demonstrated that reporter-guided mutant selection is an efficient method for further improvement of randomly mutagenized industrial strains.
A photoswitchable fluorescent protein for hours-time-lapse and sub-second-resolved super-resolution imaging
Authors: Wazawa Tetsuichi , Noma Ryohei , Uto Shusaku , Sugiura Kazunori , Washio Takashi , Nagai Takeharu
Published: January 2021

Reversibly photoswitchable fluorescent proteins (RSFPs) are a class of fluorescent proteins whose fluorescence can be turned on and off by light irradiation. RSFPs have become essential tools for super-resolution (SR) imaging. Because most SR imaging techniques require high-power-density illumination, mitigating phototoxicity in cells due to intense light irradiation has been a challenge. Although we previously developed an RSFP named Kohinoor to achieve SR imaging with low phototoxicity, the photoproperties were insufficient to move a step further to explore the cellular dynamics by SR imaging. Here, we show an improved version of RSFP, Kohinoor2.0, which is suitable for SR imaging of cellular processes. Kohinoor2.0 shows a 2.6-fold higher fluorescence intensity, 2.5-fold faster chromophore maturation and 1.5-fold faster off-switching than Kohinoor. The analysis of the pH dependence of the visible absorption band revealed that Kohinoor2.0 and Kohinoor were in equilibria among multiple fluorescently bright and dark states, with the mutations introduced into Kohinoor2.0 bringing about a higher stabilization of the fluorescently bright states compared to Kohinoor. Using Kohinoor2.0 with our SR imaging technique, super-resolution polarization demodulation/on-state polarization angle narrowing, we conducted 4-h time-lapse SR imaging of an actin filament network in mammalian cells with a total acquisition time of 480 s without a noticeable indication of phototoxicity. Furthermore, we demonstrated the SR imaging of mitochondria dynamics at a time resolution of 0.5 s, in which the fusion and fission processes were clearly visualized. Thus, Kohinoor2.0 is shown to be an invaluable RSFP for the SR imaging of cellular dynamics.
A simple preparation method for CLEM using pre-embedding immunohistochemistry with a novel fluorescent probe and stable embedding resin
Authors: Kanemaru Takaaki , Kondo Teruyoshi , Nakamura Kei-ichiro , Morimoto Hiroyuki , Nishi Kentaro , Isobe Shin-ichiro
Published: January 2021

Correlative light and electron microscopy (CLEM) is an excellent approach for examining the cellular localization of biomolecules. Here, we developed a simple method for CLEM by combining pre-embedding immunohistochemistry with a novel fluorescent probe, namely Fluolid NS Orange, and an embedding resin called ʻDurcupan™ʼ. Specimens were embedded in Durcupan™ or LR White after immunolabeling and post-fixation using glutaraldehyde and osmium tetroxide. Next, ultrathin sections were prepared on a finder grid with navigation markers. The section of the specimen embedded in Durcupan™ was found to be more stable against electron beam irradiation than specimens embedded in LR White. A fluorescence light microscopy image and a transmission electron microscopy (TEM) image, at wide-field, and low magnification, were independently obtained with the same ultrathin section. Using the three corners between finder grid bars as landmarks, fluorescence light microscopy images were superimposed with wide-field, low-magnification TEM images to identify the region of interest, which was subsequently enlarged to ascertain cellular structures localized beneath fluorescent signals. However, the enlarged TEM images appeared blurred, and fluorescence signals had a hazy appearance. To resolve this, the enlarged TEM images were replaced by high-resolution TEM images focused directly on the region of interest, thereby facilitating the collection of high-resolution CLEM images. The simple sample processing method for CLEM using osmium-resistant Fluolid NS Orange and electron beam damage-resistant Durcupan™ allowed the determination of the precise localization of fluorescence signals at subcellular levels.
A project review under the focus of ‘complexities’ on the example of exploreAT!
Authors: Dorn Amelie , Wandl-Vogt Eveline , Palfinger Thomas , Therón Roberto , Way Andy , Abgaz Yalemisew , Benito Alejandro
Published: January 2021

This article provides insights into dealing with complexities in the Digital Humanities project exploreAT!. By exploring a non-standard language collection for cultural insights, a three-fold approach is presented looking into concrete realizations and solutions of tackling challenges in terms of Open Innovation infrastructure, technology and the topic of choice, food. Methods and processes applied and developed in the project are aimed to serve as examples for future projects with similar data sets.
Polarization of hot Jupiter systems: a likely detection of stellar activity and a possible detection of planetary polarization
Authors: Bailey Jeremy , Bott Kimberly , Cotton Daniel V , Kedziora-Chudczer Lucyna , Zhao Jinglin , Evensberget Dag , Marshall Jonathan P , Wright Duncan , Lucas P W
Published: January 2021

We present high-precision linear polarization observations of four bright hot Jupiter systems (τ Boo, HD 179949, HD 189733, and 51 Peg) and use the data to search for polarized reflected light from the planets. The data for 51 Peg are consistent with a reflected light polarization signal at about the level expected with 2.8σ significance and a false alarm probability of 1.9 per cent. More data will be needed to confirm a detection of reflected light in this system. HD 189733 shows highly variable polarization that appears to be most likely the result of magnetic activity of the host star. This masks any polarization due to reflected light, but a polarization signal at the expected level of ∼20 ppm cannot be ruled out. τ Boo and HD 179949 show no evidence for polarization due to reflected light. The results are consistent with the idea that many hot Jupiters have low geometric albedos. Conclusive detection of polarized reflected light from hot Jupiters is likely to require further improvements in instrument sensitivity.
The Mastigamoeba balamuthi Genome and the Nature of the Free-Living Ancestor of Entamoeba
Authors: Žárský Vojtěch , Klimeš Vladimír , Pačes Jan , Vlček Čestmír , Hradilová Miluše , Beneš Vladimír , Nývltová Eva , Hrdý Ivan , Pyrih Jan , Mach Jan , Barlow Lael , Stairs Courtney W , Eme Laura , Hall Neil , Eliáš Marek , Dacks Joel B , Roger Andrew , Tachezy Jan
Published: January 2021

The transition of free-living organisms to parasitic organisms is a mysterious process that occurs in all major eukaryotic lineages. Parasites display seemingly unique features associated with their pathogenicity; however, it is important to distinguish ancestral preconditions to parasitism from truly new parasite-specific functions. Here, we sequenced the genome and transcriptome of anaerobic free-living Mastigamoeba balamuthi and performed phylogenomic analysis of four related members of the Archamoebae, including Entamoeba histolytica, an important intestinal pathogen of humans. We aimed to trace gene histories throughout the adaptation of the aerobic ancestor of Archamoebae to anaerobiosis and throughout the transition from a free-living to a parasitic lifestyle. These events were associated with massive gene losses that, in parasitic lineages, resulted in a reduction in structural features, complete losses of some metabolic pathways, and a reduction in metabolic complexity. By reconstructing the features of the common ancestor of Archamoebae, we estimated preconditions for the evolution of parasitism in this lineage. The ancestor could apparently form chitinous cysts, possessed proteolytic enzyme machinery, compartmentalized the sulfate activation pathway in mitochondrion-related organelles, and possessed the components for anaerobic energy metabolism. After the split of Entamoebidae, this lineage gained genes encoding surface membrane proteins that are involved in host–parasite interactions. In contrast, gene gains identified in the M. balamuthi lineage were predominantly associated with polysaccharide catabolic processes. A phylogenetic analysis of acquired genes suggested an essential role of lateral gene transfer in parasite evolution (Entamoeba) and in adaptation to anaerobic aquatic sediments (Mastigamoeba).
An Update of Neuroanesthesia for Intraoperative Brain Mapping Craniotomy
Authors: Lee Chanhung Z. , Poon Clara C. M.
Published: January 2021

The perioperative multidisciplinary team approach has probably been best exemplified by the care of awake craniotomy patients. Advancement in anesthesia and meticulous perioperative care has supported the safety and complexity of the surgical and mapping efforts in glioma resection. The discussions in this review will emphasize on anesthetic and perioperative management strategies to prevent complications and minimize their effects if they occur, including current practice guidelines in anesthesia, updates on the applications of anesthetic medications, and emerging devices. Planning the anesthetic and perioperative management is based on understanding the pharmacology of the medications, the goals of different stages of the surgery and mapping, and anticipating potential problems.
Gorgocephalidae (Digenea: Lepocreadioidea) in the Indo-West Pacific: new species, life-cycle data and perspectives on species delineation over geographic range
Authors: Huston Daniel C , Cutmore Scott C , Miller Terrence L , Sasal Pierre , Smit Nico J , Cribb Thomas H
Published: January 2021

The digenetic trematode family Gorgocephalidae comprises just a few species, and the literature devoted to the lineage consists of only a handful of reports. With one exception, all reports have been based on material collected in the Indo-West Pacific, an expansive marine ecoregion stretching from the east coast of Africa to Easter Island, Hawaii and French Polynesia. We collected adult and intramolluscan gorgocephalids from kyphosid fishes and littorinid gastropods from several Australian localities, and from South Africa and French Polynesia. Specimens of Gorgocephalus kyphosi and G. yaaji were collected from, or near, their type-localities, providing new morphological and molecular (COI, ITS2 and 28S) data needed for a revised understanding of species boundaries in the family. Two new species are recognized: Gorgocephalus euryaleae sp. nov. and Gorgocephalus graboides sp. nov. New definitive host records are provided for described species and three new intermediate hosts are identified. These new records are all associated with Kyphosus fishes and littorinid gastropods, reaffirming the restriction of gorgocephalids to these hosts. Most significantly, we provide evidence that G. yaaji is distributed from South Africa to French Polynesia, spanning the breadth of the Indo-West Pacific. Our findings have significant relevance regarding digenean species delineation over geographic range.
Antibacterial activity of the ethyl acetate part of
Abrus cantoniensis
against
Staphylococcus aureus
Authors: Nie Xu , Liu Huifang , Xu Xinyi , Zhan Yonghua , Xie Hui , Zeng Qi
Published: January 2021

Objective:

The aim of this work was to measure the antibacterial activity (against
Escherichia coli
and
Staphylococcus aureus
[
S. aureus
]) of the ethyl acetate part of
Abrus cantoniensis
and assess their potential as medicines.

Methods:

The experiment was divided into four groups: negative control group [with Mueller-Hinton broth (MHB)], positive control group (with 75% ethanol), blank group (with MHB) and test group (with the ethyl acetate part of
Abrus cantoniensis
). The antibacterial activities of the extracts were evaluated by the Oxford cup assay and minimum inhibitory concentration (MIC). Time-kill curve experiments, scanning electron microscopy, the content of DNA, RNA and protein were used to study the antibacterial mechanism of the ethyl acetate extract part on the growth and viability of
S. aureus
. The study procedures were approved by the Animal Care and Use Committee of Xi’an Jiaotong University (approval No. XJTULAC2016-412) on January 22, 2016.

Results:

The ethyl acetate part of
Abrus cantoniensis
extract exhibited the highest inhibitory activity against the growth of
S. aureus
with an inhibition zone diameter of 16.4 mm and MIC value of 0.5 μg/mL. The general activity range of the ethyl acetate part, determined using a time-killing curve, was found to be 0.5 μg/mL to 40 μg/mL (MIC to 80 × MIC). Changes in the scanning electron microscopy images and of DNA, RNA and proteins of
S. aureus
indicated possible mechanisms of the inhibitory activity of the ethyl acetate part.

Conclusion:

The ethyl acetate part of
Abrus cantoniensis
damaged bacterial cell structures, which results in protoplasm leakage, and eventually bacterial cell death.
The Diagnostic Utility of RAS Q61R Mutation-specific Immunohistochemistry in Epithelial-Myoepithelial Carcinoma
Authors: Nakaguro Masato , Tanigawa Maki , Hirai Hideaki , Yamamoto Yoshinari , Urano Makoto , Takahashi Reisuke H. , Sukeda Aoi , Okumura Yuki , Honda Shogo , Tasaki Koichiro , Shimizu Akira , Tsukahara Kiyoaki , Tada Yuichiro , Matsubayashi Jun , Faquin William C. , Sadow Peter M. , Nagao Toshitaka
Published: January 2021

Epithelial-myoepithelial carcinoma (EMC) is a rare salivary gland cancer characterized by biphasic tubular structures composed of inner ductal and outer clear myoepithelial cells. Because of its histologic variety and overlap of histologic features with other salivary gland tumors, there are broad differential diagnoses. The HRAS Q61R mutation has been reported to be frequent in and specific to EMC. We evaluated the usefulness of RAS Q61R mutant-specific immunohistochemical (IHC) staining for detecting this genetic alteration in EMC. We investigated 83 EMC cases and 66 cases of salivary gland tumors with an EMC-like component, including pleomorphic adenoma, adenoid cystic carcinoma, basal cell adenoma/adenocarcinoma, and myoepithelial carcinoma. Sanger sequencing was performed for HRAS, KRAS, and NRAS. The diffuse and membranous/cytoplasmic RAS Q61R IHC expression was observed in 65% of EMC cases, in which all cases harbored the HRAS Q61R mutation. IHC-positive cases were present only in de novo EMCs (54/76 cases, 71%) but not in EMCs ex pleomorphic adenoma. The immunoreactivity was almost always restricted to the myoepithelial cells. Conversely, all EMC cases lacking the HRAS Q61R mutation were negative on IHC. In addition, only 3% of EMC-like tumors showed the abovementioned immunopositivity. None of the cases examined carried KRAS or NRAS mutations. IHC for RAS Q61R is highly sensitive and specific for detecting the HRAS Q61R mutation in EMC. Since significant immunopositivity was almost exclusively identified in nearly two thirds of EMCs but seldom in the histologic mimics, the IHC of RAS Q61R is a useful tool for diagnosing EMC in general pathology laboratories.