bluuebunny/crossref_metadata_2025 huggingface dataset:
KRAS interaction with RAF1 RAS-binding domain and cysteine-rich domain provides insights into RAS-mediated RAF activation
Authors: Tran Timothy H. , Chan Albert H. , Young Lucy C. , Bindu Lakshman , Neale Chris , Messing Simon , Dharmaiah Srisathiyanarayanan , Taylor Troy , Denson John-Paul , Esposito Dominic , Nissley Dwight V. , Stephen Andrew G. , McCormick Frank , Simanshu Dhirendra K.
Published: August 2020

A vital first step of RAF activation involves binding to active RAS, resulting in the recruitment of RAF to the plasma membrane. To understand the molecular details of RAS-RAF interaction, we solved crystal structures of wild-type and oncogenic mutants of KRAS complexed with the RAS-binding domain (RBD) and the membrane-interacting cysteine-rich domain (CRD) from the N-terminal regulatory region of RAF1. Our structures revealed that RBD and CRD interact with each other to form one structural entity in which both RBD and CRD interact extensively with KRAS. Mutation at the KRAS-CRD interface resulted in a significant reduction in RAF1 activation despite only a modest decrease in binding affinity. Combining our structures and published data, we provide a model of RAS-RAF complexation at the membrane, and molecular insights into RAS-RAF interaction during the process of RAS-mediated RAF activation.
Force-dependent stimulation of RNA unwinding by SARS-CoV-2 nsp13 helicase
Authors: Mickolajczyk Keith J. , Shelton Patrick M.M. , Grasso Michael , Cao Xiaocong , Warrington Sara E. , Aher Amol , Liu Shixin , Kapoor Tarun M.
Published: August 2020

The superfamily-1 helicase non-structural protein 13 (nsp13) is required for SARS-CoV-2 replication, making it an important antiviral therapeutic target. The mechanism and regulation of nsp13 has not been explored at the single-molecule level. Specifically, force-dependent unwinding experiments have yet to be performed for any coronavirus helicase. Here, using optical tweezers, we find that nsp13 unwinding frequency, processivity, and velocity increase substantially when a destabilizing force is applied to the dsRNA, suggesting a passive unwinding mechanism. These results, along with bulk assays, depict nsp13 as an intrinsically weak helicase that can be potently activated by picoNewton forces. Such force-dependent behavior contrasts the known behavior of other viral monomeric helicases, drawing stronger parallels to ring-shaped helicases. Our findings suggest that mechanoregulation, which may be provided by a directly bound RNA-dependent RNA polymerase, enables on-demand helicase activity on the relevant polynucleotide substrate during viral replication.
Synaptic inputs to the inferior olive from cerebellar and vestibular nuclei have distinct release kinetics and neurotransmitters
Authors: Turecek Josef , Regehr Wade G.
Published: August 2020

The inferior olive (IO) is comprised of electrically-coupled neurons that make climbing fiber synapses onto Purkinje cells. Neurons in different IO subnuclei are inhibited by synapses with wide ranging release kinetics. Inhibition can be exclusively synchronous, asynchronous, or a mixture of both. Whether the same boutons, neurons or sources provide these kinetically distinct types of inhibition was not known. We find that the deep cerebellar nuclei (DCN) and vestibular nuclei (VN) are two major sources of inhibition to the IO that are specialized to provide inhibitory input with distinct kinetics. DCN to IO synapses lack fast synaptotagmin isoforms, release neurotransmitter asynchronously, and are exclusively GABAergic. VN to IO synapses contain fast synaptotagmin isoforms, release neurotransmitter synchronously, and are mediated by combined GABAergic and glycinergic transmission. These findings indicate that VN and DCN inhibitory inputs to the IO are suited to control different aspects of IO activity.
Transcriptional Atlas of Ileal-Anal Pouch Immune Cells from Ulcerative Colitis Patients
Authors: Devlin Joseph C. , Axelrad Jordan , Hine Ashley M. , Chang Shannon , Sarkar Suparna , Lin Jian-Da , Ruggles Kelly V. , Hudesman David , Cadwell Ken , Loke P’ng
Published: August 2020

How the human intestinal immune system is distinctly organized to respond to inflammation is still poorly understood. Here, we used single-cell RNA-sequencing to examine lamina propria CD45+ hematopoietic cells from patients with inflammatory bowel disease that have undergone ileal pouch-anal anastomosis, or the colon mucosa of ulcerative colitis patients. We identified a population ofIL1B+ antimicrobial macrophages andFOXP3/+BATF+ T cells that are associated and expanded in inflamed tissues, which we further validated in other scRNA-seq datasets from IBD patients. CD8+ T cells were unexpectedly more abundant in the pouch than colon. Cell type specific markers obtained from single-cell RNA-sequencing was used to infer representation from bulk RNA sequencing datasets, which further implicated antimicrobial macrophages expressingIL1BwithS100A8/A9calprotectin as being associated with inflammation, as well asBacteroidesandEscherichiabacterial species. Finally, we find that non-responsiveness to anti-integrin biologic therapies in UC patients is associated with the signature of this antimicrobial macrophage population in a subset of patients. This study identified conserved and distinct features of intestinal inflammation between parts of the small and large intestine undergoing similar inflammation conditions.
Structure of a type IV CRISPR-Cas effector complex
Authors: Zhou Yi , Bravo Jack P.K. , Taylor Hannah N. , Steens Jurre , Jackson Ryan N. , Staals Raymond H.J. , Taylor David W.
Published: August 2020

We reveal the structure of a type IV-B CRISPR effector (Csf) complex at 3.9 Å resolution using cryo-electron microscopy. The complex resembles the type III-A CRISPR Csm effector complex, but lacks subunits for RNA processing and target DNA cleavage, and is surprisingly assembled upon heterogeneous non-CRISPR RNA. These findings provide the first glimpse into the assembly and function of enigmatic type IV CRISPR systems.
New Pathways of Mutational Change in SARS-CoV-2 Proteomes Involve Regions of Intrinsic Disorder Important for Virus Replication and Release
Authors: Tomaszewski Tre , DeVries Ryan S. , Dong Mengyi , Bhatia Gitanshu , Norsworthy Miles D. , Zheng Xuying , Caetano-Anolles Gustavo
Published: August 2020

The massive worldwide spread of the SARS-CoV-2 virus is fueling the COVID-19 pandemic. Since the first whole-genome sequence was published in January 2020, a growing database of tens of thousands of viral genomes has been constructed. This offers opportunities to study pathways of molecular change in the expanding viral population that can help identify molecular culprits of virulence and virus spread. Here we investigate the genomic accumulation of mutations at various time points of the early pandemic to identify changes in mutationally highly active genomic regions that are occurring worldwide. We used the Wuhan NC_045512.2 sequence as a reference and sampled 15,342 indexed sequences from GISAID, translating them into proteins and grouping them by month of deposition. The per-position amino acid frequencies and Shannon entropies of the coding sequences were calculated for each month, and a map of intrinsic disorder regions and binding sites was generated. The analysis revealed dominant variants, most of which were located in loop regions and on the surface of the proteins. Mutation entropy decreased between March and April of 2020 after steady increases at several sites, including the D614G mutation site of the spike (S) protein that was previously found associated with higher case fatality rates and at sites of the NSP 12 polymerase and the NSP13 helicase proteins. Notable expanding mutations include R203K and G204R of the nucleocapsid (N) protein inter-domain linker region and G251V of the viroporin encoded by ORF3a between March and April. The regions spanning these mutations exhibited significant intrinsic disorder, which was enhanced and decreased by the N-protein and viroporin 3a protein mutations, respectively. These results predict an ongoing mutational shift from the spike and replication complex to other regions, especially to encoded molecules known to represent major β-interferon antagonists. The study provides valuable information for therapeutics and vaccine design, as well as insight into mutation tendencies that could facilitate preventive control.
An evolutionary perspective of DNA methylation patterns in skeletal tissues using a nonhuman primate model of osteoarthritis
Authors: Housman Genevieve , Quillen Ellen E. , Stone Anne C.
Published: August 2020

Objective

Epigenetic factors, such as DNA methylation, play an influential role in the development of the degenerative joint disease osteoarthritis (OA). These molecular mechanisms have been heavily studied in humans, and although OA affects several other animals in addition to humans, few efforts have taken an evolutionary perspective. This study explores the evolution of OA epigenetics by assessing the relationship between DNA methylation variation and knee OA development in baboons (Papio spp.) and by comparing these findings to human OA epigenetic associations.

Methods

Genome-wide DNA methylation patterns were identified in bone and cartilage of the right distal femora from 56 pedigreed, adult baboons (28 with and 28 without knee OA) using the Illumina Infinium MethylationEPIC BeadChip.

Results

Several significantly differentially methylated positions (DMPs) and regions (DMRs) were found between tissue types. Substantial OA-related differential methylation was also identified in cartilage, but not in bone, suggesting that cartilage epigenetics may be more influential in OA than bone epigenetics. Additionally, some genes containing OA-related DMPs overlap with and display methylation patterns similar to those previously identified in human OA, revealing a mixture of evolutionarily conserved and divergent OA-related methylation patterns in primates.

Conclusions

Overall, these findings reinforce current etiological perspectives of OA and enhance our evolutionary understanding of epigenetic mechanisms associated with OA. This work further establishes baboons as a valuable nonhuman primate model of OA, and continued investigations in baboons will help to disentangle the molecular mechanisms contributing to OA and their evolutionary histories.
The Naturalist's Journals of Gilbert White: exploring the roots of accounting for biodiversity and extinction accounting
Authors: Atkins Jill , Maroun Warren
Published: August 2020

Purpose

This paper explores the historical roots of accounting for biodiversity and extinction accounting by analysing the 18th-century Naturalist's Journals of Gilbert White and interpreting them as biodiversity accounts produced by an interested party. The authors aim to contribute to the accounting history literature by extending the form of accounting studied to include nature diaries as well as by exploring historical ecological accounts, as well as contributing to the burgeoning literature on accounting for biodiversity and extinction accounting.

Design/methodology/approach

The authors’ method involves analysing the content of Gilbert White's Naturalist's Journals by producing an 18th-century biodiversity account of species of flora and fauna and then interpretively drawing out themes from the Journals. The authors then provide a Whitean extinction account by comparing current species' status with White's biodiversity account from 250 years ago.

Findings

This paper uses Gilbert White's Naturalist's Journals as a basis for comparing biodiversity and natural capital 250 years ago with current species' status according to extinction threat and conservation status. Further the paper shows how early nature diary recording represents early (and probably the only) forms of accounting for biodiversity and extinction. The authors also highlight themes within White's accounts including social emancipation, problematisation, aesthetic elements and an example of an early audit of biodiversity accounting.

Research limitations/implications

There are limitations to analysing Gilbert White's Naturalist's Journals given that the only available source is an edited version. The authors therefore interpret their data as accounts which are indicative of biodiversity and species abundance rather than an exactly accurate account.

Practical implications

From the authors’ analysis and reflections, the authors suggest that contemporary biodiversity accounting needs to incorporate a combination of narrative, data accounting and pictorial/aesthetic representation if it is to provide a rich and accurate report of biodiversity and nature. The authors also suggest that extinction accounting should draw on historical data in order to demonstrate change in natural capital over time.

Social implications

Social implications include the understanding gleaned from the authors’ analysis of the role of Gilbert White as a nature diarist in society and the contribution made over time by his Journals and other writings to the development of nature accounting and recording, as well as to one’s understanding and knowledge of species of flora and fauna.

Originality/value

To the authors’ knowledge this is the first attempt to analyse and interpret nature diaries as accounts of biodiversity and extinction.
Deception by commission or by omission – management forecasts surrounding share repurchases
Authors: Chen Anthony , Lu Hung-Yuan (Richard)
Published: August 2020

Purpose

In this study, the authors extend upon Brockmanet al.(2008), who provide evidence that managers opportunistically accelerate bad news prior to share repurchases, but provide limited evidence that managers withhold good news until after repurchases. The authors examine management forecasts surrounding share repurchases in periods when companies must disclose detailed repurchase information. The authors argue these disclosures increase managers' legal and reputation risks of accelerating bad news, but have a lesser effect on delaying good news.

Design/methodology/approach

First, the authors examine whether managers alter the information released to the market before buying back shares by comparing managerial forecasts made within 30 days before the beginning of a repurchasing period with those made outside of this window. Second, the authors examine whether managers are more likely to provide good news forecasts, in terms of both magnitude and frequency, after buying back shares. Lastly, the authors examine the impact of CEO stock ownership on managerial forecasting behavior surrounding share buybacks.

Findings

Consistent with the authors’ hypotheses and contrary to Brockmanet al.(2008), the authors find limited evidence that the likelihood or magnitude of bad news forecasts is greater in the period before share buybacks. Instead, the authors document that the frequency and magnitude of good news forecasts increase in periods following share buybacks and that these associations are positively moderated by managerial equity incentives. The authors also find that the withholding of good news is associated with lower average repurchase prices and greater repurchase volume. The authors further show that, when litigation risk is greater, managers are less likely to accelerate bad news prior to repurchases and more likely to withhold good news until after. Overall, the study results are consistent with managers balancing the benefits of opportunistic repurchase behavior with the costs.

Originality/value

This study contributes to the management forecast and share repurchase literatures by providing evidence consistent with managers opportunistically releasing earnings forecasts in the period after buying back shares. Most importantly, the authors show that after the rule revision, managers refrain from actively disclosing bad news that carry higher legal costs. Instead, they opt for the omission of good news to repurchase stocks at lower prices. The study results reconcile the conflicting evidence of Brockmanet al.(2008) and Ge and Lennox (2011).
Industry 4.0 adoption key factors: an empirical study on manufacturing industry
Authors: Narula Sanjiv , Prakash Surya , Dwivedy Maheshwar , Talwar Vishal , Tiwari Surendra Prasad
Published: August 2020

Purpose

This research aims to outline the key factors responsible for industry 4.0 (I4.0) application in industries and establish a factor stratification model.

Design/methodology/approach

This article identifies the factor pool responsible for I4.0 from the extant literature. It aims to identify the set of key factors for the I4.0 application in the manufacturing industry and validate, classify factor pool using appropriate statistical tools, for example, factor analysis, principal component analysis and item analysis.

Findings

This study would shed light on critical factors and subfactors for implementing I4.0 in manufacturing industries from the factor pool. This study would shed light on critical factors and subfactors for implementing I4.0 in manufacturing industries. Strategy, leadership and culture are found key elements of transformation in the journey of I4.0. Additionally, design and development in the digital twin, virtual testing and simulations were also important factors to consider by manufacturing firms.

Research limitations/implications

The proposed I4.0 factor stratification model will act as a starting point while designing strategy, adopting readiness index for I4.0 and creating a roadmap for I4.0 application in manufacturing. The I4.0 factors identified and validated in this paper will act as a guide for policymakers, researchers, academicians and practitioners working on the implementation of Industry 4.0. This work establishes a solid groundwork for developing an I4.0 maturity model for manufacturing industries.

Originality/value

The existing I4.0 literature is critically examined for creating a factor pool that further presented to experts to ensure sufficient rigor and comprehensiveness, particularly checking the relevance of subfactors for the manufacturing sector. This work is an attempt to identify and validate major I4.0 factors that can impact its mass adoption that is further empirically tested for factor stratification.
The relationship between quality of work life and work-life-balance mediating role of job stress, job satisfaction and job commitment: evidence from India
Authors: Aruldoss Alex , Kowalski Kellyann Berube , Parayitam Satyanarayana
Published: August 2020

Purpose

The purpose of this study is to investigate the relationship between quality of work-life (QWL) and work-life balance (WLB).

Design/methodology/approach

Using a structured survey instrument, this paper gathered data from 445 respondents in cosmopolitan city in southern part of India. First psychometric properties of the instrument were tested, and then hierarchical regression was used as a statistical technique for analyzing the data.

Findings

The hierarchical regression results indicated that QWL is (1) negatively related to job stress, (2) positively related to job satisfaction and (3) positively related to job commitment. The results also indicated that (1) job stress is negatively related to WLB, (2) job satisfaction is positively related to WLB and (3) job commitment is positively related to WLB. The results also show partial mediation of job stress, job satisfaction, and job commitment in the relationship between QWL and WLB.

Research limitations/implications

Since the present research is based on self-report measures, the limitations of social desirability bias and common method bias are inherent. However, sufficient care is taken to minimize these limitations. The research has implications for human resource managers in work organizations.

Practical implications

This study contributes to both practicing managers and the literature on human resource management. The study suggests that employers need to be aware of the importance of quality of work-life and work-life balance in achieving organizational effectiveness.

Social implications

The study is expected to contribute to the welfare of the society in terms of identifying the antecedents of work-life balance.

Originality/value

This study provides new insights about the effects of QWL on WLB through mediating variables. This is a conceptual model developed and tested and first of its kind in India.
Improving patient safety in two French hospitals: why teamwork training is not enough
Authors: Petit dit Dariel Odessa , Cristofalo Paula
Published: August 2020

Purpose

The persistent challenges that healthcare organizations face as they strive to keep patients safe attests to a need for continued attention. To contribute to better understanding the issues currently defying patient safety initiatives, this paper reports on a study examining the aftermath of implementing a national team training program in two hospital units in France.

Design/methodology/approach

Data were drawn from a longitudinal qualitative study analyzing the implementation of a French patient safety program aimed at improving teamwork in hospitals. Data collection took place over a four-year period (2015–2019) in two urban hospitals in France and included multiple interviews with 31 participants and 150 h of observations.

Findings

Despite explicit efforts to improve inter-professional teamwork, three main obstacles interfered with healthcare professionals' attempts at safeguarding patients: perspectival variations in what constituted “patient safety”, a paradoxical injunction to do more with less and conflicting organizational priorities.

Originality/value

This paper exposes patient safety as misleadingly consensual and identifies a lack of alignment between stakeholders in the complex system that is a hospital. This ultimately interferes with patient safety objectives and highlights that even well-equipped, frontline actors cannot achieve long-term results without more systemic organizational changes.
Government funding and fundraising: an online experiment of nonprofit leader preferences and personality
Authors: Kim Mirae , Mason Dyana P.
Published: August 2020

Purpose

Many empirical studies have focused on whether public funding leverages (crowds in) or discourages (crowds out) private giving behavior, finding mixed results. Recent studies suggest the need to examine how nonprofits adjust their fundraising efforts after experiencing cuts or increases in government funding, which can then influence donor behavior.

Design/methodology/approach

In this study, the authors conduct an online survey experiment with nonprofit managers to test how nonprofits respond to changes in government funding.

Findings

The authors find some evidence that nonprofit organizations would change their fundraising expenses when facing cuts in government funding, yet the authors also find that the change could be either to increase or decrease fundraising spending. Since decisions are made by executive directors, the study also considered how executive personality type as maximizers or satisficers may interact with institutional and environmental constraints in decision-making. When funding goals are met, executives tend to behave as “satisficers” and are unlikely to make significant changes, even when their individual personality is more consistent with being a “maximizer.”

Research limitations/implications

The authors find these results to be the reflection of the current environment in which many nonprofits operate, characterized by pressures to keep operating costs low. The results of the experiment have implications for both funding agencies and nonprofits that strive to enhance the capacity of nonprofit services.

Originality/value

This study is the first attempt to untangle the multilayered relationships between government funding, fundraising, leader preferences and personalities, and donations using an experimental approach with current nonprofit leaders.
I need to know my past: Black fifth-graders on race and racism
Authors: Walker Irenea , Russell William B.
Published: August 2020

Purpose

This research study focuses on fifth-grade African American students who attend an all-Black charter school whose administration and teachers are committed to providing Black history instruction throughout the year. To fulfill the school's mission, the teachers integrate additional resources into the curriculum that includes lessons and activities about Black history. Therefore, the study sought to answer the following question: How does learning Black history throughout the school year impact African American fifth-grade students' self-esteem and positive self-image? The authors examined student work, conducted observations and listened as the participants engaged in critical discussions about race and racism.

Design/methodology/approach

Way to Go (WTG) is a K-12 public charter school located in an urban mid-size city in Florida, with a 100% Black student population; all WTG students receive scholarships and free lunch. The 15 participants in this study self-identified as African American fifth-grade students. The authors conducted a qualitative research study that included 13 observations, an analysis of five student work samples and a focus group interview with seven students. They used interpretative phenomenology to gather African American fifth-grade students' experiences and their interpretations of these experiences (Moustakas, 1994) while acquiring information about Black history.

Findings

The themes that emerged are it's time to go, unsung heroes and Black history is exciting. In the first theme, they learned why Blacks migrated from the South to northern cities and understood why it was time for them to go. Next, they explored the history of unsung Blacks who inspired them to think about a variety of careers to pursue. Finally, they were excited to learn Black history because they understood the importance of learning this history in order to grapple with current events, and they recognized that knowledge of this history would improve their self-worth and life choices.

Originality/value

WTG charter school exemplifies what schools should attain for regarding the teaching of Black history. Since elementary school provides the foundation for learning, it is the best time to teach African American students about self-esteem and what it means to be proud of their Blackness. The fifth-graders in this study exemplified how African American students take pride in their history and have a positive sense of self-worth when taught Black history. Black history lessons and activities such as the ones utilized in WTG school will benefit African American students and contribute to their success as students.
Legislators, organizations and ties: Understanding interest group recognition in the European Parliament
Authors: IBENSKAS RAIMONDAS , BUNEA ADRIANA
Published: August 2020

What explains Members of European Parliament's (MEPs’) decisions to recognize some interest groups as relevant policy actors? Addressing this question is fundamental for understanding the role of political elites in shaping patterns of interest representation and interest groups’ role in legislative decision making. Building on theories of legislative behaviour and informational theories of legislative lobbying, we argue that MEPs give recognition to those organizations that are instrumental for achieving key political goals: re‐election, career‐progression and policy influence. The pursuit of these goals generates different patterns of MEP recognition of interest groups. We contribute to the literature in three ways. Conceptually, we propose interest group recognition as a key concept for understanding interactions and links between legislative and non‐legislative actors. We illustrate the high conceptual relevance of recognition for interest groups research while noting its conspicuous neglect in the literature. We address this gap and place the concept central stage in understanding legislators’ attention to and behaviour towards interest organizations. Theoretically, we build on a classic framework explaining legislators’ behaviour and refine it through the lenses of informational theories of legislative lobbying. We argue and show that legislators recognize organizations that enhance electoral prospects in their home Member States, and that legislator–group ideological proximity and an interest group's prominence in a specific policy field affect MEPs’ decisions to recognize some organizations as relevant actors. Our argument acknowledges the importance of the broader context in which MEPs operate and pays attention to how they react to and interact with it. Empirically, we propose an original and innovative research design to identify and measure recognition with the help of social media data. Our measurement strategy constitutes a significant improvement insofar that it reduces the challenges of measurement bias usually associated with self‐reported data generated through interviews, surveys, or the textual analysis of newspaper articles and official documents. Our research design allows using fine‐grained measures of key dependent and explanatory variables and offers the very first analysis of MEP interest group recognition that holds across decision‐making events and policy areas. We test our argument on a new dataset with 4 million observations recording the recognition of more than 7,000 organizations by 80 per cent of MEPs serving in EP8. We find that MEPs are more likely to recognize organizations from their Member State, particularly under flexible‐ and open‐list electoral institutions. MEPs are also more likely to recognize organizations that share their ideological affinities and are prominent actors in policy areas legislators specialize in.
Institutionalizing problem‐oriented policing: An evaluation of the EMUN reform in Israel
Authors: Weisburd David , Hasisi Badi , Litmanovitz Yael , Carmel Tomer , Tshuva Shani
Published: August 2020

Research Summary

In 1979 Herman Goldstein proposed a radical reform—problem‐oriented policing (POP)—which has had tremendous impact on scholars and practitioners. Even though his paper and subsequent work led to a large body of literature on how to carry out problem‐oriented policing tactics, scholars have often ignored the question of how POP can be institutionalized in police agencies. In this article, we evaluate a reform in Israel—EMUN— that attempted to institutionalize problem‐oriented policing on a national scale. Focusing on property crime, we compare three treatment stations (with high, moderate, and low crime) with control stations chosen through a systematic matching procedure. We find that there are large and significant reductions in the targeted areas (termed “polygons”) for high‐ and moderate‐property‐ crime stations as compared with the control stations. We also do not find evidence of displacement but instead evidence of significant diffusions of crime control benefits. Importantly, property crime declines occurred in these stations overall. Significant benefits were not found for the low‐crime treatment station. We attribute this to the low base rate of crimes and low resource allocation in this station.

Policy Implications

These findings suggest that the EMUN reform provides a potential model for institutionalizing problem‐oriented policing as an organizational reform. EMUN attempted to support and reinforce each of the main steps of the problem‐oriented policing model. It also developed sophisticated computer tools to aid in this process that not only supported problem‐solving efforts but also allowed for wide‐scale supervision of each stage of the POP model.
Understanding inhaled Technosphere Insulin: Results of an early randomized trial in type 1 diabetes mellitus
Authors: McGill Janet B. , Weiss Daniel , Grant Marshall , Jones Marisa C. , Kendall David M. , Hoogwerf Byron J.
Published: August 2020

Background

Technosphere Insulin (TI) is an inhaled insulin. Studies comparing TI with short‐acting insulin analogues provide important insights on efficacy, dosing, and time course of action.

Methods

Planned enrollment of 230 subjects was limited to 138 due to premature study discontinuation. The primary efficacy endpoint was a noninferiority of glycosylated hemoglobin (HbA1c) of 0.4% for TI compared with insulin lispro (LIS) in a 16‐week phase 3 randomized clinical trial in type 1 diabetes mellitus.

Results

HbA1c values were similar in the TI and LIS groups at the beginning of the trial (7.8% and 7.6%, respectively) and at trial endpoint (7.7% and 7.6%, respectively). Least squares mean changes from baseline were similar between study groups. Glucose values after a standard meal were significantly lower with TI in the first 90 minutes post meal compared with LIS. Mild or moderate hypoglycemia event rates were also significantly lower with TI compared with LIS (5.97 vs 8.01, respectively; P = .0269). Cough was the most commonly reported adverse event with TI. Pulmonary function as measured by forced expiratory volume in 1 second was not different between groups at baseline, 16 weeks, or 4 weeks off study drug.

Conclusions

HbA1c was unchanged and overall glucose control was comparable between groups. Treatment with TI resulted in improved post‐meal glucose and a lower risk of hypoglycemia compared with LIS.
Cereal asparagine synthetase genes
Authors: Raffan Sarah , Halford Nigel G.
Published: August 2020

Asparagine synthetase catalyses the transfer of an amino group from glutamine to aspartate to form glutamate and asparagine. The accumulation of free (nonprotein) asparagine in crops has implications for food safety because free asparagine is the precursor for acrylamide, a carcinogenic contaminant that forms during high‐temperature cooking and processing. Here we review publicly available genome data for asparagine synthetase genes from species of the Pooideae subfamily, including bread wheat and related wheat species (Triticum and Aegilops spp.), barley (Hordeum vulgare) and rye (Secale cereale) of the Triticeae tribe. Also from the Pooideae subfamily: brachypodium (Brachypodium dIstachyon) of the Brachypodiae tribe. More diverse species are also included, comprising sorghum (Sorghum bicolor) and maize (Zea mays) of the Panicoideae subfamily and rice (Oryza sativa) of the Ehrhartoideae subfamily. The asparagine synthetase gene families of the Triticeae species each comprise five genes per genome, with the genes assigned to four groups: 1, 2, 3 (subdivided into 3.1 and 3.2) and 4. Each species has a single gene per genome in each group, except that some bread wheat varieties (genomes AABBDD) and emmer wheat (Triticum dicoccoides; genomes AABB) lack a group 2 gene in the B genome. This raises questions about the ancestry of cultivated pasta wheat and the B genome donor of bread wheat, suggesting that the hybridisation event that gave rise to hexaploid bread wheat occurred more than once. In phylogenetic analyses, genes from the other species cluster with the Triticeae genes, but brachypodium, sorghum and maize lack a group 2 gene, while rice has only two genes, one group 3 and one group 4. This means that TaASN2, the most highly expressed asparagine synthetase gene in wheat grain, has no equivalent in maize, rice, sorghum or brachypodium. An evolutionary pathway is proposed in which a series of gene duplications gave rise to the five genes found in modern Triticeae species.
Postmortem toxicological analyses of blood samples from 107 patients receiving opioid agonist treatment: substances detected and pooled opioid and benzodiazepine concentrations
Authors: Bech Anne Berit , Clausen Thomas , Waal Helge , Vindenes Vigdis , Edvardsen Hilde Erøy , Frost Joachim , Skeie Ivar
Published: August 2020

Aims

To present the substances and their concentrations detected postmortem in patients receiving opioid agonist treatment (OAT) stratified by cause of death, estimate the pooled opioid and benzodiazepine concentrations using established conversion factors for blood concentrations from the Norwegian Road Traffic Act and explore the association between drug‐induced cause of death and the pooled opioid and benzodiazepine concentrations.

Design

Cross‐sectional nation‐wide study.

Setting

Norway.

Participants

One hundred and seven patients who died during OAT (i.e. within 5 days after the last intake of OAT medication) between 1 January 2014 and 31 December 2015, with postmortem femoral blood available for toxicology. Data were collected from hospital records, the Norwegian Cause of Death Registry and autopsy reports.

Measurements

Presence of alcohol and non‐alcohol substances in the bloodstream postmortem, determined through records of toxicology of postmortem femoral blood.

Findings

A median of four substances was detected across the causes of death. At least one benzodiazepine was detected in 81 (76%) patients. The median pooled opioid concentration was significantly higher in drug‐induced deaths compared with other causes of death (362 versus 182 ng/ml, P < 0.001), in contrast to the pooled benzodiazepine concentration (5466 versus 5701 ng/ml, P = 0.353). The multivariate regression analysis showed that only increasing pooled opioid concentration (ng/ml) was associated with increased odds of a drug‐induced cause of death (odds ratio = 1.003; 95% confidence interval = 1.001–1.006).

Conclusions

In Norway, overall opioid concentration seems to play an important role in drug‐induced deaths during opioid agonist treatment in patients prescribed methadone or buprenorphine. Patients prescribed buprenorphine tend to replace their agonist with full agonists, while patients prescribed methadone tend to have high opioid concentrations from methadone as the only opioid.
The Asp298Asn polymorphism of melanocortin‐4 receptor (MC4R) in pigs: evidence for its potential effects on MC4R constitutive activity and cell surface expression
Authors: Zhang J. , Li J. , Wu C. , Hu Z. , An L. , Wan Y. , Fang C. , Zhang X. , Li J. , Wang Y.
Published: August 2020

In humans and mice, melanocortin receptor 4 (MC4R) and melanocortin receptor accessory protein 2 (MRAP2) can form a complex and control energy balance, thus regulating body weight and obesity. In pigs, a missense variant (p.Asp298Asn) of MC4R has been suggested to be associated with growth and fatness; however, the effect of Asp298Asn substitution on MC4R function is controversial, limiting its application in animal breeding. Here we examined the effect of this polymorphism on MC4R constitutive activity, cell surface expression and signaling, and its interaction with MRAP2 in pigs. We found that: (i) both pig MC4RAsp and MC4RAsn can be activated by its ligands (α‐MSH and ACTH) and stimulate cAMP/PKA signaling pathway, as detected by pGL3–CRE–luciferase reporter assay, indicating that, like pMC4RAsp, pMC4RAsn is coupled to the cAMP/PKA signaling pathway; (ii) compared with pMC4RAsp, pMC4RAsn loses the basal constitutive activity and shows a decreased surface expression, as detected by dual‐luciferase reporter assay and Nano‐HiBiT system; (iii) as in other vertebrates, both pMC4RAsp and pMC4RAsn can interact with pMRAP2, thus decreasing receptor surface expression and enhancing ligand sensitivity, although, in contrast to pMC4RAsp, the basal constitutive activity of pMC4RAsn cannot be affected by pMRAP2; and (iv) RNA‐seq data analysis revealed a co‐expression of MC4R and MRAP2 in pig hypothalamus. Taken together, our data provide convincing evidence that Asp298Asn substitution decreases the constitutive activity and cell surface expression of MC4R or MC4R–MRAP2 complex, which may affect energy balance and be a valuable selection marker for breeding programs in pigs.
Anatomy of the dura mater venous sinus of Alouatta belzebul
Authors: Sabec Pereira Dayane Kelly , de Melo Fabiano Rodrigues , de Melo Fabiana Cristina Silveira Alves , Pereira Kleber Fernando , Vulcani Valcinir Aloisio Scalla
Published: August 2020

The anatomy of the dura mater's venous sinuses is important in the veterinary clinical and surgical area, for cranial procedures and interventions of wild animals, in addition to assisting in the management of neurological diseases, which can prevent serious complications. The macroscopic anatomy of the venous sinuses of the dura mater of Alouatta belzebul was studied. Five adult specimens, males and females, were dissected and fixed in 10% formaldehyde solution. In the dura mater of the Alouatta belzebul, nine venous sinuses were observed, being them dorsal sagittal sinus, ventral sagittal sinus, transverse sinus, straight sinus, sigmoid sinus, temporal sinus, parietal sinus, basilar sinus and cavernous sinus, with morphological similarities in origin, path and destination of blood flow to the internal jugular vein, assisting in the venous drainage function of the brain in this species. These data are similar to those found in other species of non‐human and human primates such as Saimiri sciureus, Sapajus libidinosus and Homo sapiens. Of the venous sinuses observed, the cavernous sinus was of considerable clinical and surgical importance in Alouatta belzebul due to its topographic arrangement next to the internal carotid artery and pituitary gland, data that corroborate Sapajus libidinosus, Macaca fascicularis, Macaca mulatta, Papio ursinus, Cercopithecus pygerithrus and Galago senegalensis. In this context, the knowledge of the macroscopic anatomy of the venous sinuses of the dura mater may contribute to the role of veterinarians in less invasive surgical procedures in non‐human primates such as Alouatta belzebul and other mammals.
CaMK4 promotes abortion‐related Th17 cell imbalance by activating AKT/mTOR signaling pathway
Authors: Chang Shuo , Yin Tailang , He Fan , Ding Jinli , Shang Ye , Yang Jing
Published: August 2020

Problem

The balance of the immune microenvironment along the maternal‐fetal interface is closely related to pregnancy outcomes, with excessive inflammatory reactions leading to the occurrence of pathological pregnancy outcomes such as abortion. CaMK4 has been reported to play a significant role in autoimmune diseases through the regulation of Th17 cells. However, whether CaMK4 is associated with spontaneous abortion or the immune microenvironment along the maternal‐fetal interface remains unclear.

Methods of study

In this study, we constructed normal pregnancy and LPS‐induced abortion models in mice, and a CaMK4 inhibitor called KN‐93 was administered to investigate the changes in and mechanisms of the immune response. The expression of CaMK4 was evaluated in the uteroplacental complex and spleen. Furthermore, the infiltration and function of Th17 cells were estimated in peripheral tissues and the uteroplacental complex.

Results

The expression of CaMK4 in the uteroplacental complex and spleen was significantly higher in the LPS‐treated group than in the normal pregnancy group. KN‐93, the CaMK4 inhibitor, reversed fetal resorption and excessive inflammation. In detail, KN‐93 led to reduced infiltration of Th17 cells into peripheral tissues and the uteroplacental complex, and the functions of Th17 cells were inhibited. In addition, CaMK4 promoted the AKT/mTOR signaling pathway, which is one of the mechanisms that regulate the immune microenvironment.

Conclusion

CaMK4 is a critical regulator that promotes the expansion of Th17 cells and enhances their functions through the AKT/mTOR signaling pathway. The inhibition of CaMK4 can reverse the immune imbalance along the maternal‐fetal interface and improve pregnancy outcomes.
Modeling the conversion between specific IgE test platforms for nut allergens in children and adolescents
Authors: Hoang Jennifer A. , Celik Alper , Lupinek Christian , Valenta Rudolf , Duan Lucy , Dai Ruixue , Brydges May G. , Dubeau Aimée , Lépine Claire , Wong Samantha , Alexanian‐Farr Mara , Magder Ahuva , Subbarao Padmaja , Upton Julia E. M. , Schmidthaler Klara , Szépfalusi Zsolt , Ramani Arun , Eiwegger Thomas
Published: August 2020

Background

Multiplex tests allow for measurement of allergen‐specific IgE responses to multiple extracts and molecular allergens and have several advantages for large cohort studies. Due to significant methodological differences, test systems are difficult to integrate in meta‐analyses/systematic reviews since there is a lack of datasets with direct comparison. We aimed to create models for statistical integration of allergen‐specific IgE to peanut/tree nut allergens from three IgE test platforms.

Methods

Plasma from Canadian and Austrian children/adolescents with peanut/tree nut sensitization and a cohort of sensitized, high‐risk, pre‐school asthmatics (total n = 166) were measured with three R&D multiplex IgE test platforms: Allergy Explorer version 1 (ALEX) (Macro Array Dx), MeDALL‐chip (Mechanisms of Development of Allergy) (Thermo Fisher), and EUROLINE (EUROIMMUN). Skin prick test (n = 51) and ImmunoCAP (Thermo Fisher) (n = 62) results for extracts were available in a subset. Regression models (Multivariate Adaptive Regression Splines, local polynomial regression) were applied if >30% of samples were positive to the allergen. Intra‐test correlations between PR‐10 and nsLTP allergens were assessed.

Results

Using two regression methods, we demonstrated the ability to model allergen‐specific relationships with acceptable measures of fit (r2 = 94%‐56%) for peanut and tree nut sIgE testing at the extract and molecular‐level, in order from highest to lowest: Ara h 2, Ara h 6, Jug r 1, Ana o 3, Ara h 1, Jug r 2, and Cor a 9.

Conclusion

Our models support the notion that quantitative conversion is possible between sIgE multiplex platforms for extracts and molecular allergens and may provide options to aggregate data for future meta‐analysis.
Late‐onset hypogonadism: Reductio ad absurdum of the cardiovascular risk‐benefit of testosterone replacement therapy
Authors: Sesti Franz , Pofi Riccardo , Minnetti Marianna , Tenuta Marta , Gianfrilli Daniele , Isidori Andrea M.
Published: August 2020

Background

Low testosterone (T) level is considered a marker of poor cardiovascular health. Ten years ago, the Testosterone in Older Men with Mobility Limitations (TOM) trial was discontinued due to a higher number of adverse events in men receiving T compared with placebo. Since then, several studies have investigated the risks of T replacement therapy (TRT) in late‐onset hypogonadism (LOH).

Objective

To review the mechanism by which TRT could damage the cardiovascular system.

Materials and methods

Comprehensive literature search of recent clinical and experimental studies.

Results

The mechanisms of T‐mediated coronary vasodilation were reviewed with emphasis on calcium‐activated and ATP‐sensitive potassium ion channels. We showed how T regulates endothelial nitric oxide synthase (eNOS) and phosphoinositide 3‐kinase/protein kinase B/eNOS signaling pathways in vessel walls and its direct effects on cardiomyocytes via β1‐adrenergic and ryanodine receptors and provided data on myocardial infarction and heart failure. Vascular smooth muscle senescence could be explained by the modulation of growth factors, matrix metalloproteinase‐2, and angiotensin II by T. Furthermore, leukocyte trafficking, facilitated by changes in TNF‐α, could explain some of the effects of T on atheromatous plaques. Conflicting data on prothrombotic risk linked to platelet aggregation inhibition via NO‐triggered arachidonate synthesis or increased aggregability due to enhanced thromboxane A in human platelets provide evidence regarding the hypotheses on plaque maturation and rupture risk. The effects of T on cardiac electrophysiology and oxygen delivery were also reviewed.

Discussion

The effects of TRT on the cardiovascular system are complex. Although molecular studies suggest a potential benefit, several clinical observations reveal neutral or occasionally detrimental effects, mostly due to confounding factors.

Conclusions

Attempts to demonstrate that TRT damages the cardiovascular system via systematic analysis of the putative mechanisms led to the contradiction of the initial hypothesis. Current evidence indicates that TRT is safe once other comorbidities are addressed.