bluuebunny/crossref_metadata_2025 huggingface dataset:
Alarm sounded over declining US radiation professional workforce
Authors: Kramer David
Published: October 2023

As retirements surge, shortages threaten to slow advances in cancer therapy, diagnostics, and improved understanding of the physiological impacts of radiation.
When your academic ladder is longer
Authors: Ivie Rachel , White Susan
Published: October 2023

Race and ethnicity can affect how quickly a faculty member receives tenure.
Small lakes could destabilize Earth’s ice sheets
Authors: Poinar Kristin
Published: October 2023

In Greenland and Antarctica, pools of meltwater are one of many threats to the ice sheets’ longevity.
Possible relationships between the physical conditions of cattle and the occurrence of structural modifications of their coat hair
Authors: Kawahara Yutaka , Kawahara Naoya , Yamamoto Masaki , Wakizaka Hiroyuki , Okada Michiko , Ikeda Yoshimitsu
Published: October 2023

Context Concerning the epithelial tissue in the skin, including hair, vitamin A is required for the integrity of epithelial cells, strengthened by compounding keratin fibres. Further, the complex hormonal changes seen in pregnancy may contribute to the hair growth. Thus, the structure of the cattle coat hair may change depending on the conditions of vitamin A deficiency or pregnancy. Aims The effects of several diseases, i.e. vitamin A deficiency, pneumonia, chronic pneumonia, and chronic renal failure, or pregnancy, on the structure of the coat hair were investigated. Methods Physical properties of the coat hair, collected from Japanese black cattle (Bos Taurus) and Holstein cows (Bos taurus Taurus), were investigated to study the effects of vitamin A deficiency, other diseases, or pregnancy on the formation of hair. Key results When vitamin A concentration was lowered to about half of the minimum concentration for vitamin A adequacy of 20 μg/dL in plasma (i.e. deficiency level was high), we could perceive the interference in the self-assembly of the keratin fibrils. However, we also confirmed structural modifications of the keratin fibres obtained from the cattle suffering chronic renal failure. The coat hair collected from the pregnant cows tended to show superior tensile strengths to those of the hair from the non-pregnant ones when compared at the same diameter level, although pregnancy might have increased cortisol affecting the hair follicle function. Further, from the X-ray diffraction measurements, the non-pregnant cow coat hairs, having exhibited the inferior tensile properties, were found to be structurally modified. Conclusions The structure of the coat hair could be modified when the vitamin A concentration was severely reduced (vitamin A deficiency). The physiological conditions during pregnancy may be advantageous to serve the regular formation of the hair. Implications Hair has a function serving as a proxy of health status for a certain period of animal life.
The evolution of multiple sclerosis disease-modifying therapies: An update for pharmacists
Authors: Edinger Alexandra , Habibi Mitra
Published: October 2023

Purpose

Multiple sclerosis (MS) and the evolution of disease-modifying therapies (DMTs) and their indications, mechanisms of action, efficacy, pregnancy class, and cost are discussed.

MS is an immune-mediated, demyelinating, and progressive neurological disorder that can cause both motor and cognitive deficits. Onset of MS typically occurs between the ages of 20 and 40 years, and the disease can result in significant disability over time. Since the introduction of the first DMT for the treatment of MS in 1993, significant progress has been made in the development of new classes of DMTs with different mechanisms of action, higher efficacy, and simpler administration schedules, offering patients better alternatives. However, drawbacks with the use of DMTs include their increasing cost and formulary restrictions.

Conclusion

The treatment landscape of MS has significantly changed over the past 2 decades, and the introduction of newer classes of DMTs provides an opportunity for pharmacists to play an important role in the management of this patient population.
Gains and losses of the epiphytic lifestyle in epidendroid orchids: review and new analyses of succulence traits
Authors: Collobert Géromine , Perez-Lamarque Benoît , Dubuisson Jean-Yves , Martos Florent
Published: October 2023

Background and Aims

Epiphytism has evolved repeatedly in plants and has resulted in a considerable number of species with original characteristics. Because water supply is generally erratic compared to that in soils, succulent forms in particular are widespread in epiphytic species. However, succulent organs also exist in terrestrial plants, and the question of the concomitant evolution of epiphytism and succulence has received little attention, not even in the epidendroid orchids, which account for 67.6 % of vascular epiphytes.

Methods

We built a new time-calibrated phylogenetic tree of Epidendroideae with 203 genera treated in genus Orchidacearum, from which we reconstructed the evolution of epiphytism as well as traits related to water scarcity (stem and leaf succulence and the number of velamen layers), while testing for the correlated evolution between the two. Furthermore, we estimated the ancestral geographical ranges to evaluate the palaeoclimatic context in which epiphytism evolved.

Key Results

Epiphytism evolved at least three times: 39.0 million years ago (Mya) in the common ancestor of the Malaxideae and Cymbidieae that probably ranged from the Neotropics to Southeast Asia and Australia, 11.5 Mya in the Arethuseae in Southeast Asia and Australia, and 7.1 Mya in the neotropical Sobralieae, and it was notably lost in the Malaxidiinae, Collabieae, Calypsoeae, Bletiinae and Eulophiinae. Stem succulence is inferred to have evolved once, in a terrestrial ancestor at least 4.1 Mya before the emergence of epiphytic lineages. If lost, stem succulence was almost systematically replaced by leaf succulence in epiphytic lineages.

Conclusions

Epiphytism may have evolved in seasonally dry forests during the Eocene climatic cooling, among stem-succulent terrestrial orchids. Our results suggest that the emergence of stem succulence in early epidendroids was a key innovation in the evolution of epiphytism, facilitating the colonization of epiphytic environments that later led to the greatest diversification of epiphytic orchids.
Staminal hairs increase pollinator attraction and pollination accuracy in Tradescantia fluminensis (Commelinaceae)
Authors: Li Deng-Fei , Chen Yi-Dan , Liu Zhen , Liang Ai-Ting , Tang Ju , Yan Xian-Chun
Published: October 2023

Staminal hairs are the particular appendages of stamens, which may affect pollinator foraging behaviour and pollen transfer. However, experimental evidence of the functions of staminal hairs in pollination remains scarce. Here, we conducted staminal hair manipulation experiments in Tradescantia fluminensis (Commelinaceae) to investigate their effects on visitation and pollen transfer by bees. Our observations revealed that both visitation rates and visit duration of honeybees (Apis cerana) to control flowers were significantly higher than that of hairless flowers. Moreover, removing the staminal hairs significantly decreased pollen deposition by honeybees (A. cerana), but did not affect pollen removal. The staminal hair was similar in length to the stamen and the pistil of T. fluminensis. The staminal hairs provide more footholds for honeybees, and they lay prone on the staminal hairs to collect pollen, which increased the accuracy of pollination through the consistent pollen placement and pick-up on the ventral surface of honeybees. These results showed that the staminal hairs in T. fluminensis may represent an adaptation to attract pollinators and enhance pollination accuracy.
Assessment of phenotypic heterogeneity in Salmonella Typhimurium preadapted to ciprofloxacin and tetracycline
Authors: Dawan Jirapat , Kim Songrae , Ahn Juhee
Published: October 2023

Bacterial population exposed to stressful antibiotic conditions consists of various subpopulations such as tolerant, persister, and resistant cells. The aim of this study was to evaluate the phenotypic heterogeneity of Salmonella Typhimurium preadapted to sublethal concentrations of antibiotics. Salmonella Typhimurium cells were treated with 1/2 × MIC of antibiotics for the first 48 h and successively 1 × MIC for the second 24 h at 37°C, including untreated control (CON), no antibiotic and 1 × MIC ciprofloxacin (NON-CIP), 1/2 × MIC ciprofloxacin and 1 × MIC ciprofloxacin (CIP-CIP), 1/2 × MIC tetracycline and 1 × MIC ciprofloxacin (TET-CIP), no antibiotic and 1 × MIC tetracycline (NON-TET), 1/2 × MIC ciprofloxacin and 1 × MIC tetracycline (CIP-TET), and 1/2 × MIC tetracycline and 1 × MIC tetracycline (TET-TET). All treatments were evaluated by antibiotic susceptibility, ATP level, relative fitness, cross-resistance, and persistence. S. Typhimurium cells were more susceptible to non-adapted NON-CIP and NON-TET (>3-log reduction) than pre-adapted CIP-CIP, TET-CIP, CIP-TET, and TET-TET. CON exhibited the highest ATP level, corresponding to the viable cell number. The relative fitness levels were more than 0.95 for all treatments, except for NON-CIP (0.78). The resistance to ciprofloxacin and tetracycline was increased at all treatments with the exception of NON-TET. The persister cells were noticeably induced at CIP-TET treatment, showing more than 5 log CFU mL−1. The results suggest that the antibiotic preadaptation led to heterogeneous populations including persisters that can develop to resistance. This study provides new insight in the bacterial persistence associated with their potential risk and paves the way to design antibiotic therapy targeting dormant bacteria.
Associations of Blood Absolute Neutrophil Count and Cytokines With Cognitive Function in Dementia-Free Participants: A Population-Based Cohort Study
Authors: Fa Wenxin , Liang Xiaoyan , Liu Keke , Wang Nan , Liu Cuicui , Tian Na , Zhu Min , Ma Yixun , Song Lin , Tang Shi , Cong Lin , Wang Yongxiang , Hou Tingting , Du Yifeng , Qiu Chengxuan
Published: October 2023

Background

The relationships of neutrophils and cytokines with cognitive dysfunction are poorly defined. We aimed to investigate the association of peripheral blood absolute neutrophil count (ANC) with cognitive function in older adults and to further explore the mediating role of serum cytokines in this association.

Methods

This population-based cohort study included 1 666 dementia-free participants (age ≥60 years) derived from baseline examinations (March–September 2018) of the Multimodal Intervention to Delay Dementia and Disability in Rural China (MIND-China); of these, 1 087 participants completed follow-up examinations in October–December 2019. We used a neuropsychological test battery to assess episodic memory, verbal fluency, attention, and executive function at the baseline and follow-up examinations. We used Mindray BC-6800 automated hematology analyzer to measure ANC and Meso Scale Discovery to measure serum interleukin-6 (IL-6) and eotaxin-3.

Results

The linear regression analysis of cross-sectional data at baseline (n = 1 666) suggested that increased ANC was significantly associated with a lower episodic memory z score (β coefficient: −0.149, 95% CI: −0.274 to −0.023) and lower long-delayed free recall z score (−0.216, −0.361 to −0.070). Serum IL-6 and eotaxin-3 could mediate 16.16% to 20.21% and 7.55% to 9.35%, respectively, of these associations. The analysis of longitudinal data (n = 1 087) showed a J-shaped relationship of ANC with decline in episodic memory z score (p for nonlinear = .049), and a U-shaped relationship between ANC and decline in long-delayed free recall z score (p for nonlinear = .043).

Conclusions

Increased neutrophils are associated with poor cognitive performance and accelerated decline in episodic memory, and the cross-sectional association is partly mediated by serum cytokines.
International parliamentary institutions and political donations: the case of the European Parliament
Authors: Katsaitis Alexander
Published: October 2023

International Parliamentary Institutions (IPIs) increasingly welcome non-state actors (NSAs) within their policy-making procedures. Political donations are an important tool in NSAs' lobbying toolbox that has been well researched at the national level. Nevertheless, we know little about the links between NSAs, IPIs and political finance. Employing the European Parliament as an exploratory case-study, the article meticulously assesses micro-level financial donor data. Contrasting neo-functional and intergovernmental approaches, the results demonstrate a cleavage formed between individuals supporting anti-integration political parties, and international business donating to pro-integration parties. Theoretically, the article validates postfunctionalist approaches as a model for parts of the NSA–IPI relationship. Conceptually, the analysis shows that IPIs can encourage transnational cleavage formation and serve a representative function, which addresses the concerns about democratic deficit that come with the transfer of policy-making authority to international organizations. Simultaneously, financial donations can strengthen movements that oppose an IPI, and have an asymmetric impact on the IPI's political legitimacy. This calls for further research on NSAs' political financing of IPIs, the impact of financial donations on IPIs and the need to update relevant regulatory frameworks.
Sweden, NATO and the role of diasporas in foreign policy
Authors: Aggestam Lisbeth , Schierenbeck Isabell , Wackenhut Arne
Published: October 2023

When and how do diasporas influence the foreign policy of liberal democratic states? Few studies have sought to conceptualize how diasporas affect strategic bilateral relations between states. We argue that these non-state actors are an increasingly important factor in western liberal democratic societies, which challenge traditional theories of foreign policy. To explore when and how the transnational societal ties and interests of diaspora groups affect foreign and security relations between states, we develop a triadic analytical model of state–diaspora interactions and specify key contextual factors. To illustrate the dynamics at play, we analyse the influence of the Kurdish diaspora in Sweden on Turkish–Swedish bilateral relations in the wake of Sweden's decision to apply for full NATO membership. Our case-study builds on semi-structured interviews as well as news media sources, speeches and official documents. The study shows that the Kurdish diaspora demonstrates an independent role and an ability to shape the policy process due to the permeability of the political system. As a result, the policy-process is becoming more domesticated, thereby constraining the role of the executive. The article contributes important insights to policy-makers and diplomats on what impact transnational state–society relations can have on foreign policymaking.
Trust and calculation in international negotiations: how trust was lost after Brexit
Authors: Wille Tobias , Martill Benjamin
Published: October 2023

How does a loss of trust affect international negotiations? We know that trust enables cooperation and comes with the risk of betrayal. Yet the effects of lost trust are not well understood, with most research in International Relations (IR) focusing on how trust is initially built in adversarial relations. In this article, we conceptualize trust as the eclipsing of possible futures in which others do not behave cooperatively. When trust is lost, actors include further possibilities of non-cooperative behavior in their calculations and adjust their negotiating demands accordingly. We use this framework to explain the European Commission's response to Boris Johnson's government breaking its trust during the Brexit negotiations by deviating from the Political Declaration, introducing the Internal Market Bill, and not conducting border checks in the Irish Sea. Drawing on elite interviews, we trace how the Commission altered its expectations and consequently sought negotiated outcomes with tighter contracting, reduced discretion and retaliatory mechanisms. Analysing the interplay of trust and calculation on a granular level, we show that breaches of trust do not preclude future cooperation per se, but shape its form and extent in important ways.
Identification of the immunogenic membrane proteins, catalase, PgbA, and PgbB, as potential antigens against Helicobacter pylori
Authors: Li Zhirong , Zhang Ying , Mi Chaoyi , Deng Xiaoqing , Wang Xian , Hu Dailun , Yin Kaige , Yin Changfu , Zhao Lianmei , Shan Baoen
Published: October 2023

Aims

This study aims to investigate the specific membrane antigens that are targeted by antibodies raised against Helicobacter pylori.

Methods and results

Bovine milk antibodies were prepared using whole H. pylori, purified membrane proteins, or both. Enzyme-linked immunosorbent assay and sodium dodecyl sulfate–polyacrylamide gel electrophoresis experiments revealed that these immunogens triggered anti-H. pylori antibody production in milk. The highest antibody titer was induced by the mixture of whole bacteria and purified membrane proteins. The antibodies induced by mixed immunogens significantly inhibited H. pylori growth in vitro and were used to identify catalase, plasminogen-binding protein A (PgbA), and PgbB via western blotting, immunoprecipitation, and two-dimensional western blotting followed by liquid chromatography with tandem mass spectrophotometry. The immunogenicity of PgbA and PgbB was verified in mice vaccinated with their B-cell epitope vaccines. Following prophylactic vaccination of C57BL/6 mice, each of the three antigens alone and their combination reduced the weight loss in mice, increased antibody titers, and relieved the inflammatory status of the gastric mucosa following H. pylori infection.

Conclusions

Catalase, PgbA, and PgbB could serve as valuable membrane antigens for the development of anti-H. pylori immunotherapies.
Genomic selection for survival under naturally occurring Saprolegnia oomycete infection in farmed European whitefish Coregonus lavaretus
Authors: Calboli Federico C F , Iso-Touru Terhi , Bitz Oliver , Fischer Daniel , Nousiainen Antti , Koskinen Heikki , Tapio Miika , Tapio Ilma , Kause Antti
Published: October 2023

Saprolegnia oomycete infection causes serious economic losses and reduces fish health in aquaculture. Genomic selection based on thousands of DNA markers is a powerful tool to improve fish traits in selective breeding programs. Our goal was to develop a single nucleotide polymorphism (SNP) marker panel and to test its use in genomic selection for improved survival against Saprolegnia infection in European whitefish Coregonus lavaretus, the second most important farmed fish species in Finland. We used a double digest restriction site associated DNA (ddRAD) genotyping by sequencing method to produce a SNP panel, and we tested it analyzing data from a cohort of 1,335 fish, which were measured at different times for mortality to Saprolegnia oomycete infection and weight traits. We calculated the genetic relationship matrix (GRM) from the genome-wide genetic data, integrating it in multivariate mixed models used for the estimation of variance components and genomic breeding values (GEBVs), and to carry out Genome-Wide Association Studies for the presence of quantitative trait loci (QTL) affecting the phenotypes in analysis. We identified one major QTL on chromosome 6 affecting mortality to Saprolegnia infection, explaining 7.7% to 51.3% of genetic variance, and a QTL for weight on chromosome 4, explaining 1.8% to 5.4% of genetic variance. Heritability for mortality was 0.20 to 0.43 on the liability scale, and heritability for weight was 0.44 to 0.53. The QTL for mortality showed an additive allelic effect. We tested whether integrating the QTL for mortality as a fixed factor, together with a new GRM calculated excluding the QTL from the genetic data, would improve the accuracy estimation of GEBVs. This test was done through a cross-validation approach, which indicated that the inclusion of the QTL increased the mean accuracy of the GEBVs by 0.28 points, from 0.33 to 0.61, relative to the use of full GRM only. The area under the curve of the receiver–operator curve for mortality increased from 0.58 to 0.67 when the QTL was included in the model. The inclusion of the QTL as a fixed effect in the model increased the correlation between the GEBVs of early mortality with the late mortality, compared to a model that did not include the QTL. These results validate the usability of the produced SNP panel for genomic selection in European whitefish and highlight the opportunity for modeling QTLs in genomic evaluation of mortality due to Saprolegnia infection.
Sgo1 interacts with CENP-A to guide accurate chromosome segregation in mitosis
Authors: Wu Fengge , Akbar Hameed , Wang Chunyue , Yuan Xiao , Dou Zhen , Mullen McKay , Niu Liwen , Zhang Liang , Zang Jianye , Wang Zhikai , Yao Xuebiao , Song Xiaoyu , Liu Xing
Published: October 2023

Shugoshin-1 (Sgo1) is necessary for maintaining sister centromere cohesion and ensuring accurate chromosome segregation during mitosis. It has been reported that the localization of Sgo1 at the centromere is dependent on Bub1-mediated phosphorylation of histone H2A at T120. However, it remains uncertain whether other centromeric proteins play a role in regulating the localization and function of Sgo1 during mitosis. Here, we show that CENP-A interacts with Sgo1 and determines the localization of Sgo1 to the centromere during mitosis. Further biochemical characterization revealed that lysine and arginine residues in the C-terminal domain of Sgo1 are critical for binding CENP-A. Interestingly, the replacement of these basic amino acids with acidic amino acids perturbed the localization of Sgo1 and Aurora B to the centromere, resulting in aberrant chromosome segregation and premature chromatid separation. Taken together, these findings reveal a previously unrecognized but direct link between Sgo1 and CENP-A in centromere plasticity control and illustrate how the Sgo1–CENP-A interaction guides accurate cell division.
Hepatic angiomyolipoma, misdiagnosed as hepatocellular carcinoma
Authors: Marinho Bárbara M , Canha António G , Silva Donzília S , Rodrigues Ana P
Published: October 2023

Perivascular epithelioid cell neoplasm (PEComa) is a rare type of tumor, and hepatic PEComa is even rarer. Its preoperative diagnosis is difficult, given the absence of specific clinical manifestations, often constituting an accidental finding, and the lack of a gold standard for identification using imaging studies. Instead, the diagnosis of hepatic PEComa is based on morphological and immunohistochemical features. We describe a case of an asymptomatic hepatic PEComa, angiomyolipoma type, which appeared in a middle-aged woman with chronic liver disease, during her follow-up and screening. Given the patient's context, human immunodeficiency virus-positive with chronic hepatitis C, and the similarities between the two tumors, the hepatic lesion was interpreted as hepatocellular carcinoma. The patient underwent surgical excision of the tumor, and the positive immunohistochemical staining for human melanoma black 45 and Melan A made the definitive diagnosis. In the absence of aggressiveness tumor markers, surveillance was decided. We also provide a literature review of these tumors.
Biostimulated-sesame sprout extracts as potential agents against Leishmania mexicana
Authors: Garduño-Félix Karime G , Rochín-Medina Jesús J , Murua-López Carolina C , López-Moreno Héctor S , Ramírez Karina
Published: October 2023

Leishmania mexicana is one of the causal agents of cutaneous leishmaniasis. Current antileishmanial chemotherapeutics have demonstrated adverse side effects; thus, alternative treatments are needed. In this study, we performed in silico and in vitro analyses of the leishmanicidal potential of the most abundant phenolic compounds identified in black sesame sprouts biostimulated with Bacillus clausii. The molecular docking analysis showed strong interactions (binding free energies between −6.5 and −9.5 kcal/mol) of sesaminol 2-O-triglucoside, pinoresinol dihexoside, isoverbascoside, and apigenin with the arginase, leishmanolysin, cysteine peptidase B, and pyruvate kinase leishmanial enzymes. Furthermore, almost all phenolic compounds interacted with the active site residues of L. mexicana enzymes. In vitro, the B. clausii-biostimulated sprout phenolic extracts and apigenin inhibited the growth of promastigotes with IC50 values of 0.08 mg gallic acid equivalent/mL and 6.42 μM (0.0017 mg/mL), respectively. Additionally, in the macrophage infection model, cells treated with B. clausii-biostimulated sprout phenolic extracts and infected with L. mexicana exhibited significantly (P < 0.05) reduced nitric oxide production and decreased parasite burden. Altogether, our study provides important data related to high efficacy and less toxic natural antileishmanial candidates against promastigotes of L. mexicana.
Characterization of white rot fungi from wood decayed for lignin degradation
Authors: Nurul-Aliyaa Yakath Ali , Awang Nik Azimatolakma , Mohd Masratul Hawa
Published: October 2023

The present study was conducted to isolate and identify white rot fungi (WRF) from wood decayed and to determine their ability to produce lignin-modifying enzymes (LMEs), specifically laccase (Lac), lignin peroxidase (LiP), and manganese peroxidase (MnP), on solid and liquid media supplemented with synthetic dyes namely 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), azure B, and phenol red. A total of 23 isolates of WRF were isolated from decayed wood and identified as eight different species namely Phanerochaete australis, Perenniporia tephropora, Lentinus squarrosulus, Ganoderma australe, Trametes polyzona, Lentinus sajor-caju, Gymnopilus dilepis, and Fomitopsis palustris based on morphological characteristics, DNA sequences of the internal transcribed spacer (ITS) region, and phylogenetic inference. The fungal isolates can be divided into four groups based on the type of LMEs produced, namely A (Lac-LiP-MnP) with 16 isolates, B (Lac-MnP) (three isolates), C (Lac) (three isolates), and D (MnP) (one isolate). This study highlights P. australis (BJ38) as the best producer of Lac and LiP, while L. squarrosulus (IPS72) is the best producer of MnP. The present study is the first reported P. australis as an efficient lignin degrader by demonstrating the highest activity of two important LMEs.
Hypocalcemia and cardiovascular mortality in cinacalcet users
Authors: Goto Shunsuke , Hamano Takayuki , Fujii Hideki , Taniguchi Masatomo , Abe Masanori , Nitta Kosaku , Nishi Shinichi
Published: October 2023

Background

Calcimimetics are widely used in hemodialysis patients and influence serum calcium levels. Although the Kidney Disease: Improving Global Outcomes guidelines argued that low calcium levels induced by calcimimetics may be harmless, large observational studies investigating the association between hypocalcemia and mortality are scarce. We investigated the association between serum calcium levels and cardiovascular mortality in calcimimetics users using the nationwide Japanese registry for dialysis patients.

Methods

In this 9-year prospective cohort study, the baseline data were collected at the end of 2009. We enrolled patients on maintenance hemodialysis or hemodiafiltration. We employed three models (baseline, time-dependent and time-averaged) to conduct Cox proportional hazard regression analyses.

Results

Cinacalcet was prescribed to 12.7% (N = 22 853) at baseline. The median observation period was 98 (interquartile range 40–108) months and 108 (interquartile range 59–108) months in the whole cohort (N = 180 136) and in cinacalcet users, respectively. Three-quarters of survivors at the end of 2019 had continued calcimimetic therapy for 10 years, corresponding to a mean annual dropout rate of 2.9%. Hypocalcemia was not associated with cardiovascular mortality in the baseline or time-averaged model. In the time-dependent model, however, the lowest calcium decile (corrected calcium <8.4 mg/dL) was significantly associated with higher cardiovascular mortality than the reference (corrected calcium 8.7–8.9 mg/dL) in both cinacalcet users and all patients [hazard ratio (95% confidence interval) 1.32 (1.00, 1.75) and 1.15 (1.05, 1.26), respectively]. Hypocalcemia was especially associated with sudden death and death due to hemorrhagic stroke, heart failure and ischemic heart disease. Higher rate of fatal and non-fatal cardiovascular events was observed in hypocalcemic patients regardless of cinacalcet usage.

Conclusions

Our findings suggest that transient hypocalcemia was associated with an increased risk of cardiovascular death independent of cinacalcet usage. We should pay attention to hypocalcemia transiently induced by cinacalcet.
Gender balance and suitable positive actions to promote gender equality among healthcare professionals in neuro-oncology: The EANO positive action initiative
Authors: Le Rhun Emilie , Boele Florien , Minniti Giuseppe , Galldiks Norbert , Taphoorn Martin , Piil Karin , Rudà Roberta , Niclou Simone P , Geurts Marjolein , Preusser Matthias , Weller Michael , Short Susan C , Dirven Linda
Published: October 2023

Background

The proportion of women among healthcare and biomedical research professionals in neuro-oncology is growing. With changes in cultural expectations and work-life balance considerations, more men aspire to nonfull-time jobs, yet, leadership positions remain dominated by men.

Methods

The European Association of Neuro-Oncology (EANO) disparity committee carried out a digital survey to explore gender balance and actions suitable to promote gender equality. The survey was distributed among EANO members in 2021, with responses analyzed descriptively.

Results

In total, 262 participants completed the survey (141 women, 53.8%; median age 43). Respondents were neurosurgeons (68, 26.0%); neurologists (67, 25.6%), medical oncologists (43, 16.4%), or other healthcare or research professionals; 208 participants (79.4%) worked full-time. Positive action to enforce the role of women in neuro-oncology was deemed necessary by 180 participants (68.7%), but only 28 participants (10.7%) agreed that women only should be promoted until gender balance is reached. A majority of respondents (162, 61.8%) felt that women with an equivalent CV should be prioritized over men to reach gender balance. If in the future the balance favored women at higher positions, 112 respondents (42.7%) agreed to apply positive action for men. The top indicators considered relevant to measure gender balance were: salary for similar positions (183/228, 80.3%), paid overtime (176/228, 77.2%), number of permanent positions (164/228, 71.9%), protected time for research (161/227, 70.9%), and training opportunities (157/227, 69.2%).

Conclusions

Specific indicators may help to measure and promote gender balance and should be considered for implementation among healthcare professionals in neuro-oncology.
Revisiting the phylogeny of the scolebythid wasps (Hymenoptera: Aculeata) through Bayesian model evaluation and parsimony, with description of a new fossil family of Chrysidoidea
Authors: Lepeco Anderson , Melo Gabriel A R
Published: October 2023

Scolebythidae constitute the smallest family of Aculeata in number of extant species, but with a considerable diversity of extinct lineages. Twenty Cretaceous species are attributed to the family coming from deposits in Canada, Lebanon, Myanmar, New Jersey, and Spain. However, several of these taxa lack characters of the family groundplan. Although phylogenetic investigations have been conducted for the family based on a relatively small set of morphological characters, the substantial diversity of species from Burmese amber have not yet been accounted for in a phylogenetic perspective. In the present work, we reconstruct the phylogenetic relationships of Scolebythidae using a new morphological matrix and explore it through parsimony and Bayesian model evaluation. In Bayesian analyses, homoplasy-based partitioning outperformed anatomy-based partitioning and the unpartitioned models. Our phylogenetic results support the relocation of a few taxa previously attributed to Scolebythidae to the families Bethylidae and Chrysididae. We also describe †Chrysopsenellidae fam. nov., a new family of Chrysidoidea, based on an inclusion in Burmese amber. In all analyses the new family is recovered as closely related to Scolebythidae, with the whole clade being sister-group to Chrysididae + †Chrysobythidae. Finally, we discuss the origin of the Burmese amber biota distinctiveness.
PADI6: What we know about the elusive fifth member of the peptidyl arginine deiminase family
Authors: Williams Jack P. C. , Walport Louise J.
Published: October 2023

Peptidyl arginine deiminase 6 (PADI6) is a maternal factor that is vital for early embryonic development. Deletion and mutations of its encoding gene in female mice or women lead to early embryonic developmental arrest, female infertility, maternal imprinting defects and hyperproliferation of the trophoblast. PADI6 is the fifth and least well-characterized member of the peptidyl arginine deiminases (PADIs), which catalyse the post-translational conversion of arginine to citrulline. It is less conserved than the other PADIs, and currently has no reported catalytic activity. While there are many suggested functions of PADI6 in the early mouse embryo, including in embryonic genome activation, cytoplasmic lattice formation, maternal mRNA and ribosome regulation, and organelle distribution, the molecular mechanisms of its function remain unknown. In this review, we discuss what is known about the function of PADI6 and highlight key outstanding questions that must be answered if we are to understand the crucial role it plays in early embryo development and female fertility.

This article is part of the Theo Murphy meeting issue ‘The virtues and vices of protein citrullination’.
Deimination in epidermal barrier and hair formation
Authors: Méchin Marie-Claire , Simon Michel
Published: October 2023

Peptidylarginine deiminases (PADs) transform a protein arginine residue into the non-standard amino acid citrulline. This calcium-dependent post-translational modification of proteins is called citrullination or deimination. As described in this special issue, PADs play a role in various physiological processes, and PAD deregulations are involved in many human diseases. Three PADs are expressed in the epidermis, where their roles begin to be deciphered. PAD1 and PAD3 are involved in keratinocyte differentiation, particularly in the epidermal barrier function, keratins, filaggrin and filaggrin-related proteins being the most abundant deiminated epidermal proteins. Reduced amounts of deiminated proteins and PAD1 expression may be involved in the pathogenesis of psoriasis and atopic dermatitis, two very frequent and chronic skin inflammatory diseases. The trichohyalin/PAD3/transglutaminase three pathway is important for hair shaft formation. Mutations of the
PADI3
gene, leading to a decreased activity or abnormal localization of the corresponding isotype, are the cause of a rare hair disorder called uncombable hair syndrome, and are associated with the central centrifugal cicatricial alopecia, a frequent alopecia mainly affecting women of African ancestry.

This article is part of the Theo Murphy meeting issue ‘The virtues and vices of protein citrullination’.
Neutrophil attachment via Mac-1 (
α
M
β
2
; CD11b/CD18; CR3) integrins induces PAD4 deimination of profilin and histone H3
Authors: Neeli Indira , Moarefian Maryam , Kuseladass Jayalakshmi , Dwivedi Nishant , Jones Caroline , Radic Marko
Published: October 2023

Neutrophil adhesion to endothelia, entry into tissues and chemotaxis constitute essential steps in the immune response to infections that drive inflammation. Neutrophils bind to other cells and migrate via adhesion receptors, notably the
α
M
β
2
integrin dimer (also called Mac-1, CR3 or CD11b/CD18). Here, the response of neutrophils to integrin engagement was examined by monitoring the activity of peptidylarginine deiminase 4 (PAD4). Histone H3 deimination was strongly stimulated by manganese, an integrin-activating divalent cation, even in the absence of additional inflammatory stimuli. Manganese-induced cell attachment resulted in neutrophil swarm formation that paralleled histone deimination, whereas antibodies that impair integrin binding prevented both cell adhesion and histone deimination. Manganese treatment led to putative deimination of profilin, a protein that functions as an actin-organizing hub, as detected by two-dimensional gel electrophoresis and citrulline immunoblotting. Cl-amidine, a covalent inhibitor of PAD4, and GSK484, a specific PAD4 inhibitor, blocked profilin deimination. Neutrophil migration toward leukotriene B4 and toward synovial fluid from a rheumatoid arthritis patient were inhibited by chloramidine, thus supporting the contribution of deimination to chemotaxis. The data, based on a simplified system for integrin activation, imply a mechanism whereby integrin attachment coordinates neutrophil responses to inflammation and orchestrates deimination of nuclear and cytoskeletal proteins.

This article is part of the Theo Murphy meeting issue ‘The virtues and vices of protein citrullination’.
Small molecule activates citrullination through targeting PAD2
Authors: Zhang Xue , Shen Mengzhen , Zhu Huimin , Zhang Junjie , Yang Min , Su Kaiyan , Zhang Yirong , Fu Wei , Ke Xisong , Qu Yi
Published: October 2023

Citrullination is a post-translational modification catalysed by peptidyl arginine deiminase (PAD) enzymes, and dysregulation of protein citrullination is involved in various pathological disorders. During the past decade, a panel of citrullination inhibitors has been developed, while small molecules activating citrullination have rarely been reported so far. In this study, we screened citrullination activator using an antibody against citrullinated histone H3 (cit-H3), and a natural compound demethoxycurcumin (DMC) significantly activated citrullination. The requirement of PAD2 for DMC-activated citrullination was confirmed by a loss of function assay. Notably, DMC directly engaged with PAD2, and showed binding selectivity among PAD family enzymes. Point mutation assay indicated that residue E352 is essential for DMC targeting PAD2. Consistently, DMC induced typical phenotypes of cells with dysregulation of PAD2 activity, including citrullination-associated cell apoptosis and DNA damage. Overall, our study not only presents a strategy for rationally screening citrullination activators, but also provides a chemical approach for activating protein citrullination.

This article is part of the Theo Murphy meeting issue ‘The virtues and vices of protein citrullination’.
Co-expression of PADI isoforms during progenitor differentiation enables functional diversity
Authors: Vikhe Patil Kim , Meijer Mandy , Mak Kylie Hin-Man , Yang Wei , Falcão Ana Mendanha , Castelo-Branco Gonçalo , Genander Maria
Published: October 2023

Protein isoforms, generated through alternative splicing or promoter usage, contribute to tissue function. Here, we characterize the expression of predicted
Padi3α
and
Padi3β
isoforms in hair follicles and describe expression of
Padi2β
, a hitherto unknown PADI2 isoform, in the oligodendrocyte lineage.
Padi2β
transcription is initiated from a downstream intronic promoter, generating an N-terminally truncated, unstable, PADI2β. By contrast to the established role of the canonical PADI2 (PADI2α) (Falcao
et al
. 2019
Cell Rep
.
27
, 1090–1102.e10. (
doi:10.1016/j.celrep.2019.03.108
)), PADI2β inhibits oligodendrocyte differentiation, suggesting that PADI2 isoforms exert opposing effects on oligodendrocyte lineage progression. We localize
Padi3α
and
Padi3β
to developing hair follicles and find that both transcripts are expressed at low levels in progenitor cells, only to increase in expression concomitant with differentiation. When expressed
in vitro
, PADI3α and PADI3β are enriched in the cytoplasm and precipitate together. Whereas PADI3β protein stability is low and PADI3β fails to induce protein citrullination, we find that the enzymatic activity and protein stability of PADI3α is reduced in the presence of PADI3β. We propose that PADI3β modulates PADI3α activity by direct binding and heterodimer formation. Here, we establish expression and function of
Padi2
and
Padi3
isoforms, expanding on the mechanisms in place to regulate citrullination in complex tissues.

This article is part of the Theo Murphy meeting issue ‘The virtues and vices of protein citrullination’.
Neutrophil peptidylarginine deiminase 4 is essential for detrimental age-related cardiac remodelling and dysfunction in mice
Authors: Van Bruggen Stijn , Kraisin Sirima , Van Wauwe Jore , Bomhals Katrien , Stroobants Mathias , Carai Paolo , Frederix Liesbeth , Van De Bruaene Alexander , Witsch Thilo , Martinod Kimberly
Published: October 2023

Mice fully deficient in peptidylarginine deiminase 4 (PAD4) enzyme have preserved cardiac function and reduced collagen deposition during ageing. The cellular source of PAD4 is hypothesized to be neutrophils, likely due to PAD4's involvement in neutrophil extracellular trap release. We investigated haematopoietic PAD4 impact on myocardial remodelling and systemic inflammation in cardiac ageing by generating mice with
Padi4
deletion in circulating neutrophils under the MRP8 promoter (Ne-PAD4
−/−
), and ageing them for 2 years together with littermate controls (PAD4
fl/fl
). Ne-PAD4
−/−
mice showed protection against age-induced fibrosis, seen by reduced cardiac collagen deposition. Echocardiography analysis of structural and functional parameters also demonstrated preservation of both systolic and diastolic function with MRP8-driven PAD4 deletion. Furthermore, cardiac gene expression and plasma cytokine levels were evaluated. Cardiac genes and plasma cytokines involved in neutrophil recruitment were downregulated in aged Ne-PAD4
−/−
animals compared to PAD4
fl/fl
controls, including decreased levels of C-X-C ligand 1 (CXCL1). Our data confirm PAD4 involvement from circulating neutrophils in detrimental cardiac remodelling, leading to cardiac dysfunction with old age. Deletion of PAD4 in MRP8-expressing cells impacts the CXCL1–CXCR2 axis, known to be involved in heart failure development. This supports the future use of PAD4 inhibitors in cardiovascular disease.

This article is part of the Theo Murphy meeting issue ‘The virtues and vices of protein citrullination’.
An unbiased proteomic analysis of PAD4 in human monocytes: novel substrates, binding partners and subcellular localizations
Authors: Thomas Mekha A. , Kim Seok-Young , Curran Ashley M. , Smith Barbara , Antiochos Brendan , Na Chan Hyun , Darrah Erika
Published: October 2023

Peptidylarginine deiminase IV (PAD4) post-translationally converts arginine residues in proteins to citrullines and is implicated in playing a central role in the pathogenesis of several diseases. Although PAD4 was historically thought to be a nuclear enzyme, recent evidence has revealed a more complex localization of PAD4 with evidence of additional cytosolic and cell surface localization and activity. However, the mechanisms by which PAD4, which lacks conventional secretory signal sequences, traffics to extranuclear localizations are unknown. In this study, we show that PAD4 was enriched in the organelle fraction of monocytes with evidence of citrullination of organelle proteins. We also demonstrated that PAD4 can bind to several cytosolic, nuclear and organelle proteins that may serve as binding partners for PAD4 to traffic intracellularly. Additionally, cell surface expression of PAD4 increased with monocyte differentiation into monocyte-derived dendritic cells and co-localized with several endocytic/autophagic and conventional secretory pathway markers, implicating the use of these pathways by PAD4 to traffic within the cell. Our results suggest that PAD4 is expressed in multiple subcellular localizations and may play previously unappreciated roles in physiological and pathological conditions.

This article is part of the Theo Murphy meeting issue ‘The virtues and vices of protein citrullination’.
Peptidylarginine deiminase 4 and ADAMTS13 activity in
Staphylococcus
aureus
bacteraemia
Authors: Martens Caroline P. , Peetermans Marijke , Vanassche Thomas , Verhamme Peter , Jacquemin Marc , Martinod Kimberly
Published: October 2023

Staphylococcus aureus
infection is associated with increased levels of neutrophil extracellular traps (NETs) and von Willebrand factor (VWF), and with reduced activity of ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin type 1 motifs, member 13). Peptidylarginine deiminase 4 (PAD4) contributes to NET formation and inactivates ADAMTS13
in vitro
. The role of PADs in the dynamics of NETs, VWF and ADAMTS13 has not yet been studied. We thus aimed to assess the longitudinal evolution of NETs, PADs, VWF and ADAMTS13 activity in
S. aureus
infection. Plasma samples from
S. aureus
bacteraemia patients were longitudinally collected and analysed for NETs, PAD4/PAD2, VWF and ADAMTS13 activity. Correlation analyses with clinical data were performed. Recombinant PAD4 and
S. aureus
were assessed
in vitro
for their potential to modulate ADAMTS13 activity. Sixty-seven patients were included. Plasma levels of NETs, VWF, PAD4 and PAD2 were increased and ADAMTS13 activity was decreased. Levels of PADs were negatively correlated with ADAMTS13 activity. NETs were positively correlated with PADs, and negatively with ADAMTS13 activity.
In vitro
, recombinant PAD4 but not
S. aureus
reduced ADAMTS13 activity in plasma. Levels of PAD4 and PAD2 correlate with reduced ADAMTS13 activity, with neutrophils as the likely source of PAD activity in
S. aureus
bacteraemia.

This article is part of the Theo Murphy meeting issue ‘The virtues and vices of protein citrullination’.
Learning patterns of HIV-1 co-resistance to broadly neutralizing antibodies with reduced subtype bias using multi-task learning
Authors: Igiraneza Aime Bienfait , Zacharopoulou Panagiota , Hinch Robert , Wymant Chris , Abeler-Dörner Lucie , Frater John , Fraser Christophe
Published: October 2023

The ability to predict HIV-1 resistance to broadly neutralizing antibodies (bnAbs) will increase bnAb therapeutic benefits. Machine learning is a powerful approach for such prediction. One challenge is that some HIV-1 subtypes in currently available training datasets are underrepresented, which likely affects models’ generalizability across subtypes. A second challenge is that combinations of bnAbs are required to avoid the inevitable resistance to a single bnAb, and computationally determining optimal combinations of bnAbs is an unsolved problem. Recently, machine learning models trained using resistance outcomes for multiple antibodies at once, a strategy called multi-task learning (MTL), have been shown to achieve better performance in several cases than previous approaches. We develop a new model and show that, beyond the boost in performance, MTL also helps address the previous two challenges. Specifically, we demonstrate empirically that MTL can mitigate bias from underrepresented subtypes, and that MTL allows the model to learn patterns of co-resistance between antibodies, thus providing tools to predict antibodies’ epitopes and to potentially select optimal bnAb combinations. Our analyses, publicly available athttps://github.com/iaime/LBUM, can be adapted to other infectious diseases that are treated with antibody therapy.