bluuebunny/crossref_metadata_2025 huggingface dataset:
An Evaluation of Retro-Reflective Screens to Aid Conspicuity of Freight Trains at Passive-Control Level Crossings
Authors: Thompson James , Baldock Matthew , Stokes Christopher
Published: May 2021

Freight trains already passing through level crossings in rural areas at night can be difficult for approaching motorists to see. Crashes can occur if the crossing has ‘passive’ controls (Give way/Stop signs) and motorists fail to stop. Retroreflective screens on the far side of the crossing to motorists that reflect headlights and produce a ‘strobing’ effect between carriages could increase train conspicuity. A prototype screen was applied to a crossing in South Australia. Four videos of freight trains at night from the perspective of an approaching vehicle (conditions: high versus low beam headlights, screen versus no screen) were recorded and used in a reaction time experiment with N=29 drivers. Mean reaction times to the four videos were examined using multivariate analysis of variance. Results were mixed. With high beam headlights, the screen led to shorter reaction times, which suggests it increased train visibility. With low beam headlights, it led to longer reaction times, which suggests it reduced train visibility or that it confused drivers. The detrimental effect of the screen with low beam headlights could be, at least partly, due to methodological limitations relating to differences between trains in the videos, the instructions given to participants, and the degree to which the experiment replicated real-world driver behaviour. However, the screen may genuinely have confused or distracted participants and may do so in realworld conditions. Further experimental testing would be required to determine whether the results in low beam conditions persist when potential methodological limitations are addressed.
Türkakımı Projesinin Hukuki İncelemesi
Authors: DINLEYICI Ahmet
Published: May 2021

Elektrik üretiminde doğalgazın kullanımının artması ve LNG’nin birçok ülke bakımından henüz ucuz bir seçenek olmamasından dolayı gaz boru hattı projeleri önemini korumaya devam etmektedir. Her ne kadar boru hattı projeleri dünya genelinde yaygın bir biçimde yapılmaya ve uygulanmaya devam etse de, uluslararası boru hattı yasal rejimi konusunda buna mukabil bir gelişme görülmemektedir. En yeni boru hattı projelerinden biri olan ve yasal altyapısı Türkiye ve Rusya arasındaki hükümetler arası anlaşma, ev sahibi hükümet anlaşması ve diğer uluslararası anlaşmaların bazı hükümlerinden oluşan Türkakımı projesi de bu anlamda yekpare bir yasal rejim eksikligini en açık biçimde göstermektedir. Her bir projenin kendine ait bir yasal rejiminin olmasından kaynaklı farklılık ve tutarsızlıkların önüne geçmek amacıyla Enerji Şartı Sekreterligi örnek hükümetler arası anlaşma hazırlamıştır. Bu makalede, işbu örnek anlaşma metni esas olmak üzere çeşitli organizasyon raporları, kitaplar ve dergi makaleleri incelenerek Türkakımı projesi, eksik ve güçlü yönlerini tespit edebilmek amacıyla hukuki açıdan ele alınmıştır. Tüm bu araştırmaların sonucunda Türkakımı projesinin hukuksal olarak sağlam ve örnek anlaşma ile çoğunlukla örtüşmekte olduğu; ancak bazı hükümlerin eksikliğinin projenin gelecekte uygulanması aşamasında birtakım sorunlara yol açabileceği tespit edilmiştir.
Police and Judicial Cooperation in Criminal Matters in the European Union
Authors: OKUYUCU ERGÜN Güneş
Published: May 2021

Cooperation in criminal matters in the EU is probably one of the most delicate issues in the area of harmonisation as it deals with two very sensitive matters, namely, the sovereignty of Member States and the protection of human rights. Despite such delicacy, the police and judicial cooperation in criminal matters has been one of the fastest-growing areas of the EU law especially after the Treaty of Lisbon where certain competences in criminal matters were conferred to the EU. This paper concentrates on the main areas for police and judicial cooperation and the types of available legal instruments. This paper suggests that increasing the cooperation in an effective and sustainable manner would require a balanced approach between the security and the protection of fundamental rights.
Bir Sapma Davranışı Olarak Fuhuş
Authors: ARTANTAŞ Günce
Published: May 2021

Bütün hukuk düzenlerinde ortak bir suç olarak öngörülmemesi sebebiyle fuhuş, bu çalışmada bir sapma davranışı olarak nitelendirilmiş ve suçu açıklamakta da kullanılan kriminolojik teoriler ekseninde değerlendirilmiştir. Kingsley Davis’in işlevselci teorisi bu teoriler içinde önemli bir yere sahiptir. Zira bir yandan görünüşte fuhuşu kınayan cinsel ahlakın, diğer yandan ironik bir biçimde ve belki farkında olmaksızın fuhuşun sosyal bir kurum olarak varlığını sürdürmesini sağladığını ortaya koymuştur. Bu alanda ön plana çıkan bir diğer görüş ise radikal feminist teoridir. Söz konusu teoriye göre fuhuşu bir meslek olarak kabul etmenin zorluğu, insan onuruna ve vücuduna yüklenen anlamdan kaynaklanmaktadır. Fuhuşun yasal kabul edildiği Hollanda ve Yeni Zelanda gibi ülkelerde kadınların ekonomik, duygusal ya da cinsel sömürüsünün engellenemediği görülmektedir. Fuhuşu yasal hale getirmek, aslında kişi özgürlüğü adına hukuk düzeninin kişinin organlarını satmasına izin vermesinden pek de farklı değildir. Fuhuşla ilgili olarak farklı modelleri benimseyen ülkelerin yasal düzenlemeleri incelendiğinde fuhuşu bir insan hakları sorunu olarak değerlendiren, bu itibarla seks satın almayı suç haline getiren ve bedenini satmak zorunda kalan kadını ise mağdur olarak nitelendirerek ona destek sağlayan İsveç modeli, bütün ülkelerin benimsemesi gereken en modern ve hümanist çözüm olarak karşımıza çıkmaktadır.
O amor romântico no epicentro da violência doméstica contra a mulher no contexto da cultura da agressão
Authors: Gnoato Gilberto
Published: May 2021

Este artigo tem como objetivo a revisão de temáticas implicadas nos estudos da violência doméstica contra a mulher e pretende contribuir para o avanço das pesquisas de gênero, sobretudo aquelas conduzidas pelo reducionismo da “analítica do poder” (FOUCAULT, 1984). As mulheres são sempre vítimas da violência dos homens, mas isto não é tudo. Para além da teoria do patriarcado e da judicialização da intimidade, uma revisão histórico-cultural sobre o conceito de amor é fundamental para o entendimento das malhas que enredam a própria mulher no sofrimento amoroso. Adotamos a expressão “casais violentos”, segundo Machado e Magalhães (1998), analisada pela perspectiva dialógica-relacional de Santos e Izumino (2005). Destaca-se a pequena importância dada pelas pesquisas feministas, acerca do discurso do amor-paixão ou romântico nos referenciais analisados. Desta revisão, conclui-se que, uma categoria específica de mulheres, espera muito mais do amor e dependem mais dele do que esperam os homens. Utilizamos como instrumento de análise quantitativa e qualitativa, a pesquisa da antropóloga Gregori (1993) e do psicoterapeuta Gnoato (2019). Em busca de uma interdisciplinaridade, o mirante teórico deste artigo guiou-se pelo diálogo da Antropologia, com a Filosofia da Linguagem e a Psicanálise, fazendo um percurso que vai da sondagem minúscula da intimidade do casal, para uma imbricação macroscópica da cultura da agressão no Brasil, tremendamente violento, emotivo, hierárquico e paradoxal.
Biomechanical-Based Protocol for in vitro Study of Cartilage Response to Cyclic Loading: A Proof-of-Concept in Knee Osteoarthritis
Authors: Caravaggi Paolo , Assirelli Elisa , Ensini Andrea , Ortolani Maurizio , Mariani Erminia , Leardini Alberto , Neri Simona , Belvedere Claudio
Published: May 2021

Osteoarthritis (OA) is an evolving disease and a major cause of pain and impaired mobility. A deeper understanding of cartilage metabolism in response to loading is critical to achieve greater insight into OA mechanisms. While physiological joint loading helps maintain cartilage integrity, reduced or excessive loading have catabolic effects. The main scope of this study is to present an original methodology potentially capable to elucidate the effect of cyclic joint loading on cartilage metabolism, to identify mechanisms involved in preventing or slowing down OA progression, and to provide preliminary data on its application. In the proposed protocol, the combination of biomechanical data and medical imaging are integrated with molecular information about chondrocyte mechanotransduction and tissue homeostasis. The protocol appears to be flexible and suitable to analyze human OA knee cartilage explants, with different degrees of degeneration, undergoing ex vivo realistic cyclic joint loading estimated via gait analysis in patients simulating mild activities of daily living. The modulation of molecules involved in cartilage homeostasis, mechanotransduction, inflammation, pain and wound healing can be analyzed in chondrocytes and culture supernatants. A thorough analysis performed with the proposed methodology, combining in vivo functional biomechanical evaluations with ex vivo molecular assessments is expected to provide new insights on the beneficial effects of physiological loading and contribute to the design and optimization of non-pharmacological treatments limiting OA progression.
The Effects of Artificial Lake Space on Satisfaction and Restorativeness of the Overall Environment and Soundscape in Urban Parks
Authors: Qi Ying , Fang Xingyue , Gao Tian , Qiu Ling
Published: May 2021

Several studies have proven that soundscape in blue space is conducive to human health and well-being, but few studies have explored which blue space characteristics would contribute to a better soundscape and visiting experience. Therefore, an on-site questionnaire investigation was conducted at two artificial lakes in Xi’an, China. The eight Perceived Sensory Dimensions (PSDs) as a landscape assessment tool were applied to identify the characteristics of artificial lake space in urban parks. The results showed that (1) In artificial lake space, overall environment and soundscape reached a very satisfactory level in general, while the respondents’ perceived level of overall restorativeness and soundscape restorativeness as just medium, which indicated that the quality of artificial lake space needs to be improved. (2) According to people’s perceptions, artificial lake spaces had the most obvious characteristics of prospect, social and space; serene and nature were medium; refuge, rich in species, and culture were the least. (3) The eight PSDs of artificial lake space, except for social, were positively correlated with soundscape satisfaction, overall satisfaction, soundscape restorativeness, and overall restorativeness. Moreover, among them, serene was the most significant characteristic in artificial lake space. These findings could be instructive to the design of urban parks with artificial lakes for improving users’ visiting satisfaction and restorativeness.
Biological Characteristics of Umbilical Cord Mesenchymal Stem Cells and Its Therapeutic Potential for Hematological Disorders
Authors: Shang Yufeng , Guan Haotong , Zhou Fuling
Published: May 2021

Umbilical cord mesenchymal stem cells (UC-MSCs) are a class of multifunctional stem cells isolated and cultured from umbilical cord. They possessed the characteristics of highly self-renewal, multi-directional differentiation potential and low immunogenicity. Its application in the field of tissue engineering and gene therapy has achieved a series of results. Recent studies have confirmed their characteristics of inhibiting tumor cell proliferation and migration to nest of cancer. The ability of UC-MSCs to support hematopoietic microenvironment and suppress immune system suggests that they can improve engraftment after hematopoietic stem cell transplantation, which shows great potential in treatment of hematologic diseases. This review will focus on the latest advances in biological characteristics and mechanism of UC-MSCs in treatment of hematological diseases.
Circular RNA Circ0021205 Promotes Cholangiocarcinoma Progression Through MiR-204-5p/RAB22A Axis
Authors: Tu Jianfei , Chen Weiqian , Zheng Liyun , Fang Shiji , Zhang Dengke , Kong Chunli , Yang Yang , Qiu Rongfang , Zhao Zhongwei , Lu Chenying , Lu Xiaojie , Ji Jiansong
Published: May 2021

Cholangiocarcinomas (CCA) are biliary tract tumors that are often challenging to diagnosis and treatment. Accumulated evidence reveals that circular RNAs (circRNAs) are involved in multiple cancer progression. However, the function of circRNAs in cholangiocarcinoma remains largely unclear. In this study, we found that circ_0021205 expression was up-regulated in CCA and positively correlated with tumor size and TNM stage. To further explore the role of circ_0021205 in CCA, cell functional assays were performed. The results showed that circ_0021205 promoted the proliferation, migration, and invasion of CCA cells. In vivo experiments showed that circ_0021205 inhibition reduced tumorigenesis in mice. In addition, mechanisms investigation demonstrated that circ_0021205 exerts its oncogenic function by sponging miR-204-5p to regulate the expression of RAB22A. Overall, this study revealed that circ_0021205 might serve as a potential diagnostic biomarker or therapeutic target for CCA.
Oocyte Meiotic Competence in the Domestic Cat Model: Novel Roles for Nuclear Proteins BRD2 and NPM1
Authors: Chavez Daniela R. , Lee Pei-Chih , Comizzoli Pierre
Published: May 2021

To participate in fertilization and embryo development, oocytes stored within the mammalian female ovary must resume meiosis as they are arrested in meiotic prophase I. This ability to resume meiosis, known as meiotic competence, requires the tight regulation of cellular metabolism and chromatin configuration. Previously, we identified nuclear proteins associated with the transition from the pre-antral to the antral follicular stage, the time at which oocytes gain meiotic competence. In this study, the objective was to specifically investigate three candidate nuclear factors: bromodomain containing protein 2 (BRD2), nucleophosmin 1 (NPM1), and asparaginase-like 1 (ASRGL1). Although these three factors have been implicated with folliculogenesis or reproductive pathologies, their requirement during oocyte maturation is unproven in any system. Experiments were conducted using different stages of oocytes isolated from adult cat ovaries. The presence of candidate factors in developing oocytes was confirmed by immunostaining. While BRD2 and ASRGL1 protein increased between pre-antral and the antral stages, changes in NPM1 protein levels between stages were not observed. Using protein inhibition experiments, we found that most BRD2 or NPM1-inhibited oocytes were incapable of participating in fertilization or embryo development. Further exploration revealed that inhibition of BRD2 and NPM-1 in cumulus-oocyte-complexes prevented oocytes from maturing to the metaphase II stage. Rather, they remained at the germinal vesicle stage or arrested shortly after meiotic resumption. We therefore have identified novel factors playing critical roles in domestic cat oocyte meiotic competence. The identification of these factors will contribute to improvement of domestic cat assisted reproduction and could serve as biomarkers of meiotically competent oocytes in other species.
How Pointing is Integrated into Language: Evidence From Speakers and Signers
Authors: Cooperrider Kensy , Fenlon Jordan , Keane Jonathan , Brentari Diane , Goldin-Meadow Susan
Published: May 2021

When people speak or sign, they not only describe using words but also depict and indicate. How are these different methods of communication integrated? Here, we focus on pointing and, in particular, on commonalities and differences in how pointing is integrated into language by speakers and signers. One aspect of this integration is semantic—how pointing is integrated with the meaning conveyed by the surrounding language. Another aspect is structural—how pointing as a manual signal is integrated with other signals, vocal in speech, or manual in sign. We investigated both of these aspects of integration in a novel pointing elicitation task. Participants viewed brief live-action scenarios and then responded to questions about the locations and objects involved. The questions were designed to elicit utterances in which pointing would serve different semantic functions, sometimes bearing the full load of reference (‘load-bearing points’) and other times sharing this load with lexical resources (‘load-sharing points’). The elicited utterances also provided an opportunity to investigate issues of structural integration. We found that, in both speakers and signers, pointing was produced with greater arm extension when it was load bearing, reflecting a common principle of semantic integration. However, the duration of the points patterned differently in the two groups. Speakers’ points tended to span across words (or even bridge over adjacent utterances), whereas signers’ points tended to slot in between lexical signs. Speakers and signers thus integrate pointing into language according to common principles, but in a way that reflects the differing structural constraints of their language. These results shed light on how language users integrate gradient, less conventionalized elements with those elements that have been the traditional focus of linguistic inquiry.
Case Report: Phloroglucinol-Induced Kounis Syndrome
Authors: Wu Hao-Yu , Gao Tian-Jiao , Cao Yi-Wei , You Peng-Hua
Published: May 2021

Background: Kounis syndrome is an allergy-related acute coronary syndrome that is induced by various pharmacological and environmental factors. Given that many clinicians are not aware of this condition, many cases may be underdiagnosed. We report a case of type II Kounis syndrome induced by phloroglucinol.

Case Summary: A 52-year-old man with pre-existing coronary artery stenosis presented with a 30-min history of chest pain and erythematous rash after intramuscular administration of phloroglucinol. An electrocardiogram demonstrated ST-segment elevation in leads II, III and aVF. Emergency coronary angiography revealed severe stenosis in the distal right coronary artery. Intravascular ultrasound showed plaque rupture and thrombosis, and the minimum lumen area was 3.0 mm2. A 3.5 × 38 mm stent was implanted in the distal right coronary artery. Troponin I levels were elevated. A diagnosis of type II Kounis syndrome induced by phloroglucinol was made, and the condition manifested as acute ST-segment elevation myocardial infarction.

Conclusions: Clinicians should be aware of Kounis syndrome as a possible diagnosis in a patient who presents with chest pain and allergic manifestations given that an increasing number of triggers are being reported.
Persistent Symptoms and Health Needs of Women and Men With Non-Obstructed Coronary Arteries in the Years Following Coronary Angiography
Authors: Groepenhoff Floor , Eikendal Anouk L. M. , Rittersma Z. H. Saskia , Gijsberts Crystel M. , Asselbergs Folkert W. , Hoefer Imo E. , Pasterkamp Gerard , Rutten Frans H. , Onland-Moret N. Charlotte , Den Ruijter Hester M.
Published: May 2021

Background: The prognosis of women and men with persistent anginal complaints and non-obstructed coronary arteries is impaired as compared with asymptomatic women and men. The increased healthcare burden in the hospital due to repeated coronary angiography in these women and men has been documented, yet little is known about the percentage of women and men who remain symptomatic and under care of the general practitioner in the years following a coronary angiographic outcome of non-obstructed coronary arteries.

Methods: From the Utrecht Coronary Biobank study, including individuals who underwent a coronary angiography from 2011 to 2015 (N = 2,546, 27% women), we selected women and men with non-obstructed coronary arteries (N = 687, 39% women). This population was linked to the Julius General Practitioners Network (JGPN); a database with routine care data of general practitioners. For every individual with non-obstructed coronary arteries, we selected an asymptomatic non-referred age-, sex-, and general practitioner-matched individual from the JGPN. We compared the healthcare consumption of men and women with non-obstructed coronary arteries to these matched individuals. The McNemar's test was used for pairwise comparison, and sex differences were assessed using stratified analyses.

Results: The prevalence of non-obstructed coronary arteries was higher in women as compared with men (39 vs. 23%). During a median follow-up of 7 years [IQR 6.4–8.0], 89% of the individuals with non-obstructed coronary arteries (91% women and 87% men) visited their general practitioner for one or more cardiovascular consultations. This was compared to 34% of the matched individuals (89 vs. 34%, p < 0.001). The consultations were most often for angina (equivalents) (57 vs. 11%, p < 0.001) and heart failure (10 vs. 2%, p = 0.015). In addition, they more often consulted the general practitioner for psychosocial complaints (31 vs. 15%, p = 0.005). Findings were similar for women and men.

Conclusions: A coronary angiographic outcome of non-obstructed coronary arteries is more common in women than in men. In the years following the coronary angiography, the majority of the population remains symptomatic. Both women and men with non-obstructed coronary arteries had higher health needs for angina, heart failure, and psychosocial complaints than matched asymptomatic individuals.
Appraisals of Wildlife During Restorative Opportunities in Local Natural Settings
Authors: Johansson Maria , Flykt Anders , Frank Jens , Hartig Terry
Published: May 2021

Many call for a broad approach to valuation of nature’s contribution to people, one that provides a contextualized understanding of what may be experienced as a value in different cultures, groups and settings. In the present paper we address contributions of nature to psychological well-being as realized through restorative processes during encounters with wildlife. Although restorative benefits of nature experience have received much consideration, sparse attention has been given to the role of the presence or absence of different animals in the settings investigated. The presence of a liked species may increase appreciation for and engagement with a natural setting, but fear of encountering some species may counter the desire to visit a setting with otherwise high restorative quality. This paper proposes a psychological framework for understanding how wildlife may contribute to or hinder people’s opportunities to restore in local natural settings. The framework addresses the transaction between the individual and their surroundings, making use of an appraisal theory of emotion and theories about the restorative benefits of nature experience. We focus upon encounters in landscapes shared by humans and wildlife, and we elaborate on our reasoning with scenarios from Sweden involving local people’s appraisal of wolves and roe deer. An integrated understanding of the psychological processes at work would facilitate communication and decision-making about the contribution of wildlife in nature conservation and management.
How to Promote Compliance Management in the Electricity Market? An Analysis Based on the Evolutionary Game Model
Authors: Jin Luosong , Chen Cheng , Li Yun , Wang Xiangyang , Cheng Yuanyuan
Published: May 2021

In this paper, we aim to analyze how to effectively promote compliance management in the electricity market. We construct an evolutionary game model under the two different scenarios, i.e., the scenario without governmental supervision and the scenario with governmental supervision, and explicitly describes the strategic behaviors and dynamic evolution process of power enterprises and regulators in the power market. According to the results of the evolutionary stable strategy, we find that, in the absence of governmental supervision, the long-term stable equilibrium of power enterprises' choice of strategy “Compliance” and regulatory agencies' choice of strategy “Not bribery” is hard to be realized. Only if the government effectively supervises the compliance management of the electricity market can the ideal compliance behavior of the two stakeholders be achieved. Furthermore, we conduct a simulation study to analyze the impacts of the various model parameters on the dynamic evolution process. The specific results show that the lower compliance cost, the higher psychological cost, and the larger profit loss of the power enterprises, as well as the lower inspection cost, the higher psychological cost, and the larger rewards of the regulatory agencies, can promote the formation of compliance management. Besides, the larger penalty charged by the government is also conducive to the compliance management of the electricity market.
A Timely Review of Cross-Kingdom Regulation of Plant-Derived MicroRNAs
Authors: Li Dan , Yang Jianhui , Yang Yong , Liu Jianxin , Li Hui , Li Rongfei , Cao Chunya , Shi Liping , Wu Weihua , He Kai
Published: May 2021

MicroRNAs (miRNAs) belong to a class of non-coding RNAs that suppress gene expression by complementary oligonucleotide binding to the sites in target messenger RNAs. Numerous studies have demonstrated that miRNAs play crucial role in virtually all cellular processes of both plants and animals, such as cell growth, cell division, differentiation, proliferation and apoptosis. The study of rice MIR168a has demonstrated for the first time that exogenous plant MIR168a influences cholesterol transport in mice by inhibiting low-density lipoprotein receptor adapter protein 1 expression. Inspired by this finding, the cross-kingdom regulation of plant-derived miRNAs has drawn a lot of attention because of its capability to provide novel therapeutic agents in the treatment of miRNA deregulation-related diseases. Notably, unlike mRNA, some plant miRNAs are robust because of their 3′ end modification, high G, C content, and the protection by microvesicles, miRNAs protein cofactors or plant ingredients. The stability of these small molecules guarantees the reliability of plant miRNAs in clinical application. Although the function of endogenous miRNAs has been widely investigated, the cross-kingdom regulation of plant-derived miRNAs is still in its infancy. Herein, this review summarizes the current knowledge regarding the anti-virus, anti-tumor, anti-inflammatory, anti-apoptosis, immune modulation, and intestinal function regulation effects of plant-derived miRNAs in mammals. It is expected that exploring the versatile role of plant-derived miRNAs may lay the foundation for further study and application of these newly recognized, non-toxic, and inexpensive plant active ingredients.
Predicting Drug-Disease Association Based on Ensemble Strategy
Authors: Wang Jianlin , Wang Wenxiu , Yan Chaokun , Luo Junwei , Zhang Ge
Published: May 2021

Drug repositioning is used to find new uses for existing drugs, effectively shortening the drug research and development cycle and reducing costs and risks. A new model of drug repositioning based on ensemble learning is proposed. This work develops a novel computational drug repositioning approach called CMAF to discover potential drug-disease associations. First, for new drugs and diseases or unknown drug-disease pairs, based on their known neighbor information, an association probability can be obtained by implementing the weighted K nearest known neighbors (WKNKN) method and improving the drug-disease association information. Then, a new drug similarity network and new disease similarity network can be constructed. Three prediction models are applied and ensembled to enable the final association of drug-disease pairs based on improved drug-disease association information and the constructed similarity network. The experimental results demonstrate that the developed approach outperforms recent state-of-the-art prediction models. Case studies further confirm the predictive ability of the proposed method. Our proposed method can effectively improve the prediction results.
Increased Dysfunctional and Plastic Regulatory T Cells in Idiopathic Orbital Inflammation
Authors: Chen Jingqiao , Ye Huijing , Xiao Wei , Mao Yuxiang , Ai Siming , Chen Rongxin , Lian Xiufen , Shi Lu , Wang Xing , Bi Shaowei , Yang Shenglan , Ji Xian , Zhang Te , Yang Huasheng
Published: May 2021

Background

Idiopathic orbital inflammation (IOI) is a disfiguring and vision-threatening fibroinflammatory disorder. The pathogenesis of IOI has not been elucidated. We sought to clarify the regulatory T cell (Treg) distribution and function in patients with IOI.

Methods

The frequency, phenotype and function of Tregs were identified by multicolor flow cytometry andin vitrocell functional assays. Plasma and tissue samples were obtained to investigate cytokines, chemokines and their receptors of interest by relative real-time polymerase chain reaction (PCR) and Luminex assays.

Results

Compared with healthy subjects, patients with IOI exhibited obvious increases of Tregs in peripheral blood and affected orbital tissues. Circulating Tregs from patients with IOI were significantly more polarized to a Th17-like phenotype with defective regulatory function, whereas orbit-derived Tregs were polarized to a Th2-like phenotype. Furthermore, ST2 expression levels in circulating Tregs and interleukin (IL)-33 mRNA levels in orbital tissues were decreased in IOI. IL-33 restored the suppressive function of Tregs, reduced interferon (IFN)-γ production by Tregs and decreased the activation of orbital fibroblasts (OFs) cocultured with Tregs in IOI.

Conclusion

Increased Tregs with proinflammatory and profibrotic polarization were first identified in IOI, suggesting that Treg plasticity and heterogeneity plays an essential role in IOI pathogenesis. Additionally, our study identified a regulatory effect of IL-33 on inflammation and fibrosis in IOI. Reversing the plastic TregsviaIL-33 might be a potential option for IOI patients.
Applications of Protein Microarrays in Biomarker Discovery for Autoimmune Diseases
Authors: Li Siting , Song Guang , Bai Yina , Song Ning , Zhao Jiuliang , Liu Jian , Hu Chaojun
Published: May 2021

Dysregulated autoantibodies and cytokines were deemed to provide important cues for potential illnesses, such as various carcinomas and autoimmune diseases. Increasing biotechnological approaches have been applied to screen and identify the specific alterations of these biomolecules as distinctive biomarkers in diseases, especially autoimmune diseases. As a versatile and robust platform, protein microarray technology allows researchers to easily profile dysregulated autoantibodies and cytokines associated with autoimmune diseases using various biological specimens, mainly serum samples. Here, we summarize the applications of protein microarrays in biomarker discovery for autoimmune diseases. In addition, the key issues in the process of using this approach are presented for improving future studies.
Resident Memory T Cells in Autoimmune Skin Diseases
Authors: Ryan Grace E. , Harris John E. , Richmond Jillian M.
Published: May 2021

Tissue resident memory T cells (TRM) are a critical component of the immune system, providing the body with an immediate and highly specific response against pathogens re-infecting peripheral tissues. More recently, however, it has been demonstrated that TRM cells also form during autoimmunity. TRM mediated autoimmune diseases are particularly destructive, because unlike foreign antigens, the self-antigens are never cleared, continuously activating self-reactive TRM T cells. In this article, we will focus on how TRMs mediate disease in autoimmune skin conditions, specifically vitiligo, psoriasis, cutaneous lupus erythematosus, alopecia areata and frontal fibrosing alopecia.
Clozapine Regulates Microglia and Is Effective in Chronic Experimental Autoimmune Encephalomyelitis
Authors: Ceylan Ulaş , Haupeltshofer Steffen , Kämper Laura , Dann Justus , Ambrosius Björn , Gold Ralf , Faissner Simon
Published: May 2021

Objective

Progressive multiple sclerosis is characterized by chronic inflammation with microglial activation, oxidative stress, accumulation of iron and continuous neurodegeneration with inadequate effectiveness of medications used so far. We now investigated effects of iron on microglia and used the previously identified neuroprotective antipsychotic clozapine in vitro and in chronic experimental autoimmune encephalomyelitis (EAE).

Methods

Microglia were treated with iron and clozapine followed by analysis of cell death and response to oxidative stress, cytokine release and neuronal phagocytosis. Clozapine was investigated in chronic EAE regarding optimal dosing and therapeutic effectiveness in different treatment paradigms. Animals were scored clinically by blinded raters. Spinal cords were analyzed histologically for inflammation, demyelination, microglial activation and iron accumulation and for transcription changes of regulators of iron metabolism and inflammation. Effects on immune cells were analyzed using flow cytometry.

Results

Iron impaired microglial function in vitro regarding phagocytosis and markers of inflammation; this was regulated by clozapine, reflected in reduced release of IL-6 and normalization of neuronal phagocytosis. In chronic EAE, clozapine dose-dependently attenuated clinical signs and still had an effect if applied in a therapeutic setting. Early mild sedative effects habituated over time. Histologically, demyelination was reduced by clozapine and positive effects on inflammation strongly correlated with reduced iron deposition. This was accompanied by reduced expression of DMT-1, an iron transport protein.

Conclusions

Clozapine regulates microglial function and attenuates chronic EAE, even in a therapeutic treatment paradigm. This well-defined generic medication might therefore be considered as promising add-on therapeutic for further development in progressive MS.
Cholesterol-Induced M4-Like Macrophages Recruit Neutrophils and Induce NETosis
Authors: Maretti-Mira Ana C. , Golden-Mason Lucy , Salomon Matthew P. , Kaplan Mariana J. , Rosen Hugo R.
Published: May 2021

The liver is the central organ for cholesterol synthesis and homeostasis. The effects of dietary cholesterol on hepatic injury, mainly of oxidized low-density lipoproteins (OxLDL), are not fully understood. Here, we show that the degree of cholesterol oxidation had different impacts on the global gene expression of human M2-like macrophages, with highly oxidized LDL causing the most dramatic changes. M2-like macrophages and Kupffer cells undergo M4-like polarization, decreasing the expression of important markers, such as IL10, MRC1, and CD163. These cells also displayed functional changes, with reduced phagocytic capacity, increased neutrophil recruitment, and more effective neutrophil extracellular traps (NETs) induction. Our findings provide a link between LDL oxidation and modification of peripheral and liver macrophage function.
Marine Streptomyces sp. Isolated From the Brazilian Endemic Tunicate Euherdmania sp. Produces Dihydroeponemycin and Analogs With Potent Antiglioma Activity
Authors: Furtado Luciana C. , Bauermeister Anelize , de Felicio Rafael , Ortega Raquel , Pinto Francisco das Chagas L. , Machado-Neto João Agostinho , Trivella Daniela B. B. , Pessoa Otilia D. L. , Wilke Diego V. , Lopes Norberto P. , Jimenez Paula C. , Costa-Lotufo Leticia V.
Published: May 2021

Marine natural products have emerged as an important source for drug development, notably in the field of anticancer therapy. Still, the limited effectiveness of current therapies for central nervous system tumors indicates the need to identify new therapeutic targets and also novel pharmacological agents. In this context, proteasome inhibitors are appearing as a promising new treatment for these diseases. Herein, cytotoxic extracts produced by four marine bacteria recovered from the Brazilian endemic ascidian Euherdmania sp. were screened to evaluate their potential as proteasome inhibitors. The extract from marine Streptomyces sp. BRA-346 was selected for further investigation due to the potent proteasome inhibitory activity it displayed. Bioassay-guided fractionation led to an enriched fraction (proteasome inhibition IC50 = 45 ng/mL), in which the presence of dihydroeponemycin (DHE), known for its proteasome inhibitory effect, and related compounds were annotated by mass spectrometry and further confirmed by comparison with DHE standard. Both DHE and the epoxyketone-containing fraction were evaluated in glioma cell lines, displaying high cytotoxicity in HOG and T98G cells (GI50 of 1.6 and 1.7 ng/mL for DHE, and 17.6 and 28.2 ng/mL for the BRA-346 fraction, respectively). Additional studies showed that the epoxyketone-containing fraction (at GI50 levels) led to an accumulation of ubiquitinated proteins and up-regulation of genes related to ER-stress response, suggesting treated cells are under proteasome inhibition. DHE induced similar effects in treated cells but at concentrations 25 times its GI50, suggesting that the other epoxyketone compounds in the bacteria extract derived fraction may contribute to enhance proteasome inhibition and further cellular effects in glioma cells. These findings revealed the molecular pathways modulated by this class of compounds in glioma cells and, moreover, reinforced the potential of this marine bacteria in producing a cocktail of structurally-related compounds that affect the viability of glioma cells.
Low Rate of Acquired Linezolid Resistance in Multidrug-Resistant Tuberculosis Treated With Bedaquiline-Linezolid Combination
Authors: Du Jian , Gao Jingtao , Yu Yanhong , Li Qingfeng , Bai Guanghong , Shu Wei , Gao Mengqiu , Liu Yuhong , Wang Lu , Wang Yufeng , Xue Zhongtan , Huo Fengmin , Li Liang , Pang Yu
Published: May 2021

In this retrospective study in China, we aimed to: (1) determine the prevalence of linezolid (LZD) resistance among multidrug-resistant tuberculosis (MDR-TB)-infected patients; (2) monitor for dynamic LZD susceptibility changes during anti-TB treatment; and (3) explore molecular mechanisms conferring LZD resistance. A total of 277 MDR-TB patients receiving bedaquiline (BDQ)-containing regimens in 13 TB specialized hospitals across China were enrolled in the study. LZD and BDQ susceptibility rates were determined using the minimum inhibitory concentration (MIC) method, then DNA sequences of patient isolates were analyzed using Sanger sequencing to detect mutations conferring LZD resistance. Of 277 patients in our cohort, 115 (115/277, 41.5%) with prior LZD exposure yielded 19 (19/277, 6.9%) isolates exhibiting LZD resistance. The LZD resistance rate of LZD-exposed group isolates significantly exceeded the corresponding rate for non-exposed group isolates (P = 0.047). Genetic mutations were observed in 10 (52.6%, 10/19) LZD-resistant isolates, of which a Cys154Arg (36.8%, 7/19) substitution within ribosomal protein L3 was most prevalent. Analysis of sequential positive cultures obtained from 81 LZD-treated patients indicated that cultured organisms obtained from most patients (85.2%, 69/81) retained original LZD MIC values; however, organisms cultured later from two patients exhibited significantly increased MIC values that were attributed to the rplC substitution T460C. Overall, LZD resistance was detected in 6.9% of patients of an MDR-TB cohort in China. Low rate of acquired LZD resistance was noted in MDR-TB treated with BDQ-LZD combination.
The Consumption of Energy Drinks Induces Blood-Brain Barrier Dysfunction in Wild-Type Mice
Authors: Graneri Liam , Lam Virginie , D'Alonzo Zachary , Nesbit Michael , Mamo John C. L. , Takechi Ryusuke
Published: May 2021

Energy drinks containing significant quantities of caffeine and sugar are increasingly consumed, particularly by adolescents and young adults. Chronic ingestion of energy drinks may potentially regulate vascular risk factors. This study investigated the effects of chronic ingestion of energy drinks on blood-brain barrier (BBB) integrity and neuroinflammation. Male C57BL/6J mice were maintained on water (control), MotherTM (ED), sugar-free MotherTM (sfED), or Coca ColaTM soft drink (SD) for 13 weeks. The BBB integrity and neuroinflammation were analyzed with semi-quantitative immunofluorescent microscopy. Blood pressure, plasma inflammatory cytokine levels and blood glucose were also considered. Following 13 weeks of intervention, mice treated with ED, sfED, and SD showed significant disruption of BBB. However, marked neuroinflammation was observed only in sfED group mice. The consumption of ED and sfED significantly altered the blood pressure and plasma concentrations of inflammatory cytokines, TNF-a, IL-4, IL-6, and IL-10, and both increased plasma glucose. Correlation analyses showed significant associations between BBB dysfunction and hypotension, hyperglycaemia and cytokine dyshomeostasis. The intake of energy drink, particularly the sugar free formulation, may compromise the integrity of BBB and induce neuroinflammation via hypotension, hyperglycaemia and inflammatory pathways.
The Impact of Intraoperative Frozen Section on Resection Margin Status and Survival of Patients Underwent Pancreatoduodenectomy for Distal Cholangiocarcinoma
Authors: Chen Zhiqiang , Yu Bingran , Bai Jiaping , Li Qiong , Xu Bowen , Dong Zhaoru , Zhi Xuting , Li Tao
Published: May 2021

Background

Intraoperative frozen section (FS) is broadly used during pancreaticoduodenectomy (PD) to ensure a negative margin status, but its survival benefits on obtaining a secondary R0 resection for distal cholangiocarcinoma (dCCA) is controversial and unclear.

Methods

Clinical data of 107 patients who underwent PD for dCCA was retrospectively collected and divided into different groups based on use of FS (FS and non-FS groups) and status of resection margin (pR0, sR0 and R1 groups), and clinical parameters and survival of patients were compared and analyzed accordingly.

Results

There were 50 patients in FS group with a median survival of 28 months, 57 patients in non-FS group with a median survival of 27 months. There was no statistical difference between the two groups with Kaplan-Meier survival analysis (P = 0.347). There were 98 patients in R0 group (88 in pR0 and 10 in sR0) and nine patients in R1 group, with a median survival of 29 months and 22 months respectively, which showed a better survival in R0 group than in R1 group (P = 0.006). Survival analyses between subgroups revealed difference between pR0 and R1 group (P = 0.005), while no statistical difference concerning pR0 vs. sR0 (P = 0.211) and sR0 vs. R1 groups (P = 0.262). Multivariate Cox regression analysis revealed resection margin status, pre-operative biliary drainage and lymph node invasion to be independent prognostic factors for dCCA patients.

Conclusions

Intraoperative FS should be recommended as it significantly increased the rate of R0 resection, which was positively related to a better survival. A primary R0 resection should also be encouraged and if not, a secondary R0 could be considered at the discretion of surgeons as it showed similar survival with primary R0 resection.
Hypoxia-Induced MALAT1 Promotes the Proliferation and Migration of Breast Cancer Cells by Sponging MiR-3064-5p
Authors: Shih Chung-Hsien , Chuang Li-Ling , Tsai Mong-Hsun , Chen Li-Han , Chuang Eric Y. , Lu Tzu-Pin , Lai Liang-Chuan
Published: May 2021

Hypoxia, a common process during tumor growth, can lead to tumor aggressiveness and is tightly associated with poor prognosis. Long noncoding RNAs (lncRNAs) are long ribonucleotides (>200 bases) with limited ability to translate proteins, and are known to affect many aspects of cellular function. One of their regulatory mechanisms is to function as a sponge for microRNA (miRNA) to modulate its biological functions. Previously, MALAT1 was identified as a hypoxia-induced lncRNA. However, the regulatory mechanism and functions of MALAT1 in breast cancer are still unclear. Therefore, we explored whether MALAT1 can regulate the functions of breast cancer cells through miRNAs. Our results showed the expression levels of MALAT1 were significantly up-regulated under hypoxia and regulated by HIF-1α and HIF-2α. Next, in contrast to previous reports, nuclear and cytoplasmic fractionation assays and fluorescence in situ hybridization indicated that MALAT1 was mainly located in the cytoplasm. Therefore, the labeling of MALAT1 as a nuclear marker should be done with the caveat. Furthermore, expression levels of miRNAs and RNA immunoprecipitation using antibody against AGO2 showed that MALAT1 functioned as a sponge of miRNA miR-3064-5p. Lastly, functional assays revealed that MALAT1 could promote cellular migration and proliferation of breast cancer cells. Our findings provide evidence that hypoxia-responsive long non-coding MALAT1 could be transcriptionally activated by HIF-1α and HIF-2α, act as a miRNA sponge of miR-3064-5p, and promote tumor growth and migration in breast cancer cells. These data suggest that MALAT1 may be a candidate for therapeutic targeting of breast cancer progression.
Efficacy and Safety of Apatinib Treatment for Advanced Cholangiocarcinoma After Failed Gemcitabine-Based Chemotherapy: An Open-Label Phase II Prospective Study
Authors: Zhang Ge , Gong Shuai , Pang Lina , Hou Lixia , He Wei
Published: May 2021

Purpose

As a novel small-molecule vascular endothelial growth factor receptor-2 tyrosine kinase inhibitor (VEGFR2-TKI), Methylsulfonic apatinib (apatinib) exhibits a specific antitumor effect in various solid tumors via inhibition of angiogenesis. The present study was performed to evaluate the clinical efficacy and safety of apatinib in the treatment of advanced cholangiocarcinoma after failed gemcitabine-based chemotherapy.

Patients and Methods

This was a prospective open-label phase II trial (NCT03521219). A total of 32 patients, in whom gemcitabine-based first-line chemotherapy for advanced intrahepatic cholangiocarcinoma had failed, were consecutively enrolled in a prospective, open, exploratory, and single-center clinical trial from November 2017 to November 2018. They were treated with apatinib mesylate second-line monotherapy (orally, 500 mg per day for a cycle of 28 days) until progressive disease or unacceptable toxicity. Using Response Evaluation Criteria in Solid Tumor version 1.1 (RECIST 1.1) and the Common Terminology Criteria for Adverse Events version 4.0 (NCI-CTCAE 4.0), the efficacy and adverse were evaluated, respectively. Kaplan-Meier method was used for survival analysis.

Results

Twenty-six patients were enrolled in full analysis set. At the end of follow-up, two patients were lost to follow-up, 24 of 26 patients in FAS were included in efficacy analyses. For the efficacy analysis set, the objective response rate (ORR) was 20.8% [95% confidence interval (CI): 9.24–40.47%] and the disease control rate (DCR) was 62.5% (95% CI: 112.86–387.14 days). One patient (4%) showed complete response (CR), 4 patients (17%) showed partial response (PR), 10 patients (41.7%) stable disease (SD), and 9 patients (37.5%) had progressive disease (PD). Meanwhile, apatinib therapy achieved the median progression-free survival PFS was 95 days (95% CI: 79.70–154.34 days), and the median OS was 250 days (95% CI: 112.86–387.14 days). Furthermore, univariate analysis revealed that age and tumor’s anatomic location significantly affected PFS (P < 0.05). The most common clinically adverse events (AEs) included myelosuppression (69.2%), hypertension (57.7%), proteinuria (46.2%). The AEs were mild, mainly in grade 1 or 2, and no toxicity-induced death occurred.

Conclusion

Apatinib monotherapy is an effective and promising regimen for treating patients with advanced cholangiocarcinoma who experienced failure of gemcitabine-based chemotherapy.