bluuebunny/crossref_metadata_2025 huggingface dataset:
Проблема кваліфікації статусу ОРДЛО у міжнародному праві
Authors: КРЕСІН ОЛЕКСІЙ , КРЕСІНА ІРИНА
Published: January 2021

У контексті необхідності визначення суб’єкта або суб’єктів, відповідальних за дотримання нематеріальних і матеріальних прав людини та юридичних осіб, а також за порушення цих прав на території окремих районів Донецької та Луган ської областей (ОРДЛО), щодо яких Уряд України тимчасово втратив контроль, слід точно визначити статус цієї території згідно з міжнародним правом. Цю проблему можна розглядати як похідну від кваліфікації збройного конфлікту на сході України як міжнародного чи неміжнародного (хоча існують і проміжні визначення). Але вона може розглядатись і як самостійна проблема у контексті так званого права окупації та суттєвої трансформації останнього після Другої світової війни. У такому розумінні поставлена проблема може набути не лише практичного, а й наукового значення. Зрозуміло, що в цій статті йтиметься лише про постановку, а не вирішення поставлених завдань.
Автори припускають, що внаслідок тривалого розвитку класичних норм міжнародного гуманітарного права відбулося узагальнення права окупації. Можна стверджувати про існування єдиного правового режиму незаконного контролю однієї держави щодо території іншої держави. Формами такого контролю є, власне, окупація, що передбачає офіційне визнання державою здійснення нею контролю щодо території (частини території) іншої держави, та чітко не розмежовані ефективний, всезагальний, загальний, фактичний контроль. На думку авторів, останні можна вважати єдиною формою ефективного контролю, що передбачає неоголошені дії та неофіційні за соби його здійснення, намагання представити підконтрольні військові структури і військово-адміністративні інституції як самостійних недержавних акторів міжнародних відносин.
Кваліфікація статусу ОРДЛО в міжнародному праві неоднозначна. Мінські угоди 2014 р. уникали визначення другої сторони конфлікту (перша – Україна), а в 2015 р. другою стороною названі збройні формування ОРДЛО. При цьому не вказано, кому вони підпорядковуються і хто може вважатися їхніми представниками. Резо люції Парламентської Асамблеї Ради Європи (ПАРЄ) визнають подвійний факт наявності тимчасової окупації ОРДЛО Росією і здійснення нею ефективного контролю щодо незаконних збройних формувань на цій території. Важливо, що ПАРЄ вказує на застосовність щодо збройного конфлікту в Україні й зокрема ситуації в ОРДЛО IV Женевської конвенції 1949 р. Водночас повноцінне визнання наявності міжнародного збройного конфлікту між Україною і Росією на території ОРДЛО, а також незаконного контролю Росії щодо ОРДЛО (у формі окупації, ефективного чи іншого контролю) слід очікувати в процесі розгляду низки справ у Міжнародному суді Організації Об’єднаних Націй, Міжнародному кримінальному суді, Європейському суді з прав людини.
Значення державної реєстрації для набуття права власності на нерухомість
Authors: СПАСИБО-ФАТЄЄВА ІННА
Published: January 2021

Державна реєстрація має істотне значення для набуття права власності на нерухоме майно та його захисту. В Україні існує законодавство про державну реєстрацію речових прав та їх обтяжень, яке спирається на принцип внесення, тобто запроваджує титульну негативну систему. Відповідно, право власності на нерухомість має набуватися з моменту його державної реєстрації. Належне функціонування системи державної реєстрації прав на нерухоме майно має значення не тільки для приватних, а й для публічних відносин. Оскільки придбання нерухомого майна (набуття права власності на нього) є етапним, то кожному юридичному факту в юридичному складі має приділятися своє значення. При цьому якщо нерухомість придбавається за договором, то самому договору не може приділятися самодостатнього значення для набуття права власності. Таке значення має лише державна реєстрація права на майно, що в жодному разі не принижує чи не скасовує роль договору у набутті права власності.
Метою статті є визначення моменту набуття права власності на нерухоме майно через його зв’язок із державною реєстрацією.
В Україні давно назріла необхідність визначитися зі значенням державної реєстрації взагалі і для набуття права власності на нерухоме майно зокрема. Слід послідовно рухатися тим шляхом, як усі країни, в яких введена державна реєстрація прав, яка, на відміну від державної реєстрації правочинів, має не обліковий, а правовстановлюючий характер. Тлумачення закону, зокрема й вищими судовими органами при вирішенні спорів, має співвідноситися з системою державної реєстрації прав, до якої приєдналася Україна, і виходити з сутності та ідеї державної реєстрації, яка полягає у такому: всі права на нерухомість повинні бути зареєстровані; відо-мості державної реєстрації повинні бути достовірними, що має велике значення для цивільного обороту.
Спори про право на нерухомість повинні вирішуватися у кожному конкретному випадку, спираючись на принципи правомірності, добросовісності та розумності,не встановленням судом правила про значення державної реєстрації та її ролі в набутті (моменту) права власності.
Ejemplos de modelado del comportamiento a flexión de secciones de hormigón con fibras mediante la hipótesis de fisura plana
Authors: Carmona Jacinto Ruiz , Rey Juan Rey , López Gonzalo Ruiz
Published: January 2021

Este trabajo presenta un modelo para evaluar el comportamiento a flexión de secciones de hormigón reforzado con fibras (FRC). El comportamiento a tracción del hormigón con fibras se representa mediante un modelo cohesivo a través de la ley de ablandamiento lineal incluida en el Código Modelo-2010. Como ecuación de compatibilidad se hace uso de la hipótesis de fisura plana, es decir, se asume que las caras de la fisura permanecen planas durante todo el proceso de fractura, hipótesis que ha sido contrastada recientemente mediante correlación digital de imágenes [1]. El comportamiento a compresión del hormigón se representa a través de un modelo elástico, usando la hipótesis de Navier. La apertura de fisura se evalúa a partir del momento aplicado en la sección y de la profundidad de la fisura a través de la expresión propuesta por Tada et al. [2].

El modelo reproduce el efecto de escala existente en la resistencia a flexotracción (módulo de ruptura) en hormigón y hormigón con fibras. Asimismo se define un número de fragilidad que permite caracterizar el comportamiento de las secciones tanto de hormigón en masa como FRC. La metodología presentada propone el análisis de secciones de FRC a través de diagramas que representen el momento aplicado frente a la apertura de fisura, en vez de la curvatura. Esta metodología, extensible a hormigón armado, se considera una aproximación más física al análisis de secciones fisuradas y plantea a la hipótesis de fisura plana como ecuación de compatibilidad alternativa a la hipótesis de Navier, que es la normalmente incluida en las normativas.

Referencias:

[1]        S. Gali, K.V.L. Subramaniam. 2018. Multi-linear stress-crack separation relationship for steel fiber reinforced concrete: Analytical framework and experimental evaluation, Theoretical and Applied Fracture Mechanics, 93, 33-43

[2]        Tada, H. et al. 1973. The stress analysis of cracks handbook. Del Research Corporation
Puente extradosado sobre el Río Barrow. Diseño y análisis estructural del proceso constructivo, control geométrico y asistencia técnica durante la construcción
Authors: Sanchez Sanchez Marcos
Published: January 2021

En esta comunicación se presentan los elementos de diseño asociados a la ejecución del puente sobre el rio Barrow en New Ross, Irlanda. Con dos luces principales de 230 m en una configuración de puente extradorsal de tres torres, incluyendo el paso sobre el río Barrow con un canal de navegación en activo y una longitud total de 890 m distribuida en 9 vanos, los procesos constructivos del puente han sido determinantes para la optimización del proyecto. Para ello se han empleado dos procesos constructivos diferentes:

Los viaductos de acceso en ambos lados de los vanos principales extradosados se han construido en dos fases desde el punto de vista seccional. El núcleo del cajón mediante cimbra semi rígida al suelo para el núcleo central y una segunda fase mediante un carro de alas para los voladizos transversales y losas/puntal prefabricados.

Los dos vanos principales se han construido mediante voladizos sucesivos en la pila central y frente de voladizo único en las pilas laterales, en ambos casos, con carros de sección completa. Debido a la asimetría de las torres y los planos de atirantamiento (18 cables en la pila central y 8 cables en las pilas laterales) los voladizos en cada vano no son simétricos, encontrándose el punto de cierre a 140 m de la pila central y 90 de las laterales lo que requirió especial cuidado en el diseño de las fases de cierre. Asimismo, en su máxima longitud de voladizo, la pila central alcanzó 140 m en hormigón soportado por cables extradorsales, lo que constituye un récord del mundo en esta configuración
Lo pequeño es bello: algunos ejemplos de estructuras de pequeña escala
Authors: Rey Rey Juan , Ruiz Carmona Jacinto , Vegas González Pablo
Published: January 2021

Habitualmente los proyectos más divulgados son aquellos de mayor tamaño o relevancia económica. Sin embargo, en no pocas ocasiones es en los más pequeños o de menor impacto mediático en los que se desarrollan propuestas más interesantes, arriesgadas o innovadoras. Esto puede deberse a varios factores: por una parte, en los proyectos de pequeña escala es más asumible tomar un cierto grado de riesgo en las soluciones propuestas, motivado en parte por la menor envergadura económica de lo que está en juego. Por otro lado, en este tipo de proyectos cada rincón del mismo es especial y único, descartándose así en muchas ocasiones de entrada las soluciones convencionales.Dentro del campo de la edificación, las viviendas unifamiliares son una tipología muy dada a la experimentación y a la apuesta por soluciones singulares desde el punto de vista estructural. En algunos casos las singularidades vienen de la mano de grandes luces o voladizos y en otras de su complejidad geométrica o formal. En el presente artículo se analizan algunos ejemplos de esta tipología desarrollados por Mecanismo Ingeniería, como: Casa Campos en Salobreña, Vivienda “Where Eagles Dare” en Mallorca o Casa de las Hormigas en Collado Villalba (Fig. 1). Reduciendo todavía más la escala, cabe destacar que, en algunos casos, el diseño y cálculo de una escalera puede tener mayor complejidad técnica y requerir más horas de ingeniería que el propio edificio que la alberga. Así, se presentan también algunos ejemplos desarrollados como la escalera de terrazo estructural en vivienda en Beaulieu sur Mer, escalera de Casa C o escalera de Casa Cárcavas (Fig. 2).Por último, se presentan también ejemplos de otros elementos de pequeña escala como pérgolas, marquesinas o cubiertas ligeras, los cuales son frecuentemente proyectos de alta complejidad y de elevado interés. Se incluye dentro de esta clasificación la instalación artística 1.78 de Janet Echelman en la Plaza Mayor de Madrid o el Mirador da Cova de Adegas Moure en Lugo (Fig. 3).
The Truth and Myth of the Advantages of Authoritarian Countries to COVID-19
Authors: Annaka Susumu
Published: January 2021

The COVID-19 pandemic—the worst pandemic since the Spanish flu—has dramatically changed the world, with a significant number of people still suffering and dying from the disease. Some scholars argue that the pandemic has severely damaged democratic countries, mainly because they cannot intervene in their citizens' lives, as opposed to their authoritarian counterparts. This paper reports using cross-national data for 163 countries that authoritarian countries tend to have lower COVID-19 deaths than their democratic counterparts, but stringent intervention is not necessarily a key determinant. A higher number of tests is a more critical determinant of authoritarian countries' advantage. Testing seems to be a vital tool to reduce deaths. This study uses statistical evidence to demonstrate that authoritarian countries are likely to conduct more tests leading to lower death rates. The result implies that authoritarian states can strengthen citizens' support for their government through the COVID-19 pandemic.
High Content Analysis Across Signaling Modulation Treatments for Subcellular Target Identification Reveals Heterogeneity in Cellular Response
Authors: Biswas Sayan
Published: January 2021

Cellular phenotypes on bioactive compound treatment are a result of the downstream targets of the respective treatment. Here, a computational approach is taken for downstream subcellular target identification to understand the basis of the cellular response. This response is a readout of cellular phenotypes captured from cell-painting-based light microscopy images. The readouts are morphological profiles measured simultaneously from multiple cellular organelles. Cellular profiles generated from roughly 270 diverse treatments on bone cancer cell line form the high content screen used in this study. Phenotypic diversity across these treatments is demonstrated, depending on the image-based phenotypic profiles. Furthermore, the impact of the treatments on specific organelles and associated organelle sensitivities are determined. This revealed that endoplasmic reticulum has a higher likelihood of being targeted. Employing multivariate regression overall cellular response is predicted based on fewer organelle responses. This prediction model is validated against 1,000 new candidate compounds. Different compounds despite driving specific modulation outcomes elicit a varying effect on cellular integrity. Strikingly, this confirms that phenotypic responses are not conserved that enables quantification of signaling heterogeneity. Agonist-antagonist signaling pairs demonstrate switch of the targets in the cascades hinting toward evidence of signaling plasticity. Quantitative analysis of the screen has enabled the identification of these underlying signatures. Together, these image-based profiling approaches can be employed for target identification in drug and diseased states and understand the hallmark of cellular response.
The Identification of Plasma Exosomal miR-423-3p as a Potential Predictive Biomarker for Prostate Cancer Castration-Resistance Development by Plasma Exosomal miRNA Sequencing
Authors: Guo Tianyu , Wang Yang , Jia Jing , Mao Xueying , Stankiewicz Elzbieta , Scandura Glenda , Burke Edwina , Xu Lei , Marzec Jacek , Davies Caitlin R. , Lu Jiaying Jasmin , Rajan Prabhakar , Grey Alistair , Tipples Karen , Hines John , Kudahetti Sakunthala , Oliver Tim , Powles Thomas , Alifrangis Constantine , Kohli Manish , Shaw Greg , Wang Wen , Feng Ninghan , Shamash Jonathan , Berney Daniel , Wang Liang , Lu Yong-Jie
Published: January 2021

Castration-resistant prostate cancer (CRPC) is the major cause of death from prostate cancer. Biomarkers to improve early detection and prediction of CRPC especially using non-invasive liquid biopsies could improve outcomes. Therefore, we investigated the plasma exosomal miRNAs associated with CRPC and their potential for development into non-invasive early detection biomarkers for resistance to treatment. RNA-sequencing, which generated approximately five million reads per patient, was performed to identify differentially expressed plasma exosomal miRNAs in 24 treatment-naive prostate cancer and 24 CRPC patients. RT-qPCR was used to confirm the differential expressions of six exosomal miRNAs, miR-423-3p, miR-320a, miR-99a-5p, miR-320d, miR-320b, and miR-150-5p (p = 7.3 × 10−8, 0.0020, 0.018, 0.0028, 0.0013, and 0.0058, respectively) firstly in a validation cohort of 108 treatment-naive prostate cancer and 42 CRPC patients. The most significant differentially expressed miRNA, miR-423-3p, was shown to be associated with CRPC with area under the ROC curve (AUC) = 0.784. Combining miR-423-3p with prostate-specific antigen (PSA) enhanced the prediction of CRPC (AUC = 0.908). A separate research center validation with 30 treatment-naive and 30 CRPC patients also confirmed the differential expression of miR-423-3p (p = 0.016). Finally, plasma exosomal miR-423-3p expression in CRPC patients was compared to 36 non-CRPC patients under androgen depletion therapy, which showed significantly higher expression in CRPC than treated non-CRPC patients (p < 0.0001) with AUC = 0.879 to predict CRPC with no difference between treatment-naive and treated non-CRPC patients. Therefore, our findings demonstrate that a number of plasma exosomal miRNAs are associated with CRPC and miR-423-3p may serve as a biomarker for early detection/prediction of castration-resistance.
Rbm24a Is Necessary for Hair Cell Development Through Regulating mRNA Stability in Zebrafish
Authors: Zhang Yan , Wang Yanfei , Yao Xuebo , Wang Changquan , Chen Fangyi , Liu Dong , Shao Ming , Xu Zhigang
Published: January 2021

Hair cells in the inner ear and lateral lines are mechanosensitive receptor cells whose development and function are tightly regulated. Several transcription factors as well as splicing factors have been identified to play important roles in hair cell development, whereas the role of RNA stability in this process is poorly understood. In the present work, we report that RNA-binding motif protein 24a (Rbm24a) is indispensable for hair cell development in zebrafish.Rbm24aexpression is detected in the inner ear as well as lateral line neuromasts. Albeitrbm24adeficient zebrafish do not survive beyond 9 days post fertilization (dpf) due to effects outside of the inner ear,rbm24adeficiency does not affect the early development of inner ear except for delayed otolith formation and semicircular canal fusion. However, hair cell development is severely affected and hair bundle is disorganized inrbm24amutants. As a result, the auditory and vestibular function ofrbm24amutants are compromised. RNAseq analyses identified several Rbm24a-target mRNAs that are directly bound by Rbm24a and are dysregulated inrbm24amutants. Among the identified Rbm24a-target genes,lrrc23,dfna5b, andsmpxare particularly interesting as their dysregulation might contribute to the inner ear phenotypes inrbm24amutants. In conclusion, our data suggest that Rbm24a affects hair cell development in zebrafish through regulating mRNA stability.
3D Spheroids of Umbilical Cord Blood MSC-Derived Schwann Cells Promote Peripheral Nerve Regeneration
Authors: Lin Yu-Jie , Lee Yun-Wei , Chang Che-Wei , Huang Chieh-Cheng
Published: January 2021

Schwann cells (SCs) are promising candidates for cell therapy due to their ability to promote peripheral nerve regeneration. However, SC-based therapies are hindered by the lack of a clinically renewable source of SCs. In this study, using a well-defined non-genetic approach, umbilical cord blood mesenchymal stem cells (cbMSCs), a clinically applicable cell type, were phenotypically, epigenetically, and functionally converted into SC-like cells (SCLCs) that stimulated effective sprouting of neuritic processes from neuronal cells. To further enhance their therapeutic capability, the cbMSC-derived SCLCs were assembled into three-dimensional (3D) cell spheroids by using a methylcellulose hydrogel system. The cell–cell and cell–extracellular matrix interactions were well-preserved within the formed 3D SCLC spheroids, and marked increases in neurotrophic, proangiogenic and anti-apoptotic factors were detected compared with cells that were harvested using conventional trypsin-based methods, demonstrating the superior advantage of SCLCs assembled into 3D spheroids. Transplantation of 3D SCLC spheroids into crush-injured rat sciatic nerves effectively promoted the recovery of motor function and enhanced nerve structure regeneration. In summary, by simply assembling cells into a 3D-spheroid conformation, the therapeutic potential of SCLCs derived from clinically available cbMSCs for promoting nerve regeneration was enhanced significantly. Thus, these cells hold great potential for translation to clinical applications for treating peripheral nerve injury.
Caspase-8 Inhibition Prevents the Cleavage and Degradation of E3 Ligase Substrate Receptor Cereblon and Potentiates Its Biological Function
Authors: Zhou Liang , Yu Wenjun , Jayabalan David S. , Niesvizky Ruben , Jaffrey Samie R. , Huang Xiangao , Xu Guoqiang
Published: January 2021

Cereblon (CRBN), a substrate receptor of cullin 4-RING E3 ligase (CRL4), mediates the ubiquitination and degradation of constitutive substrates and immunomodulatory drug-induced neo-substrates including MEIS2, c-Jun, CLC1, IKZF1/3, CK1α, and SALL4. It has been reported that CRBN itself could be degraded through the ubiquitin-proteasome system by its associated or other cullin-RING E3 ligases, thus influencing its biological functions. However, it is unknown whether the CRBN stability and its biological function could be modulated by caspases. In this study, using model cell lines, we found that activation of the death receptor using tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) leads to the decreased CRBN protein level. Through pharmacological inhibition and activation of caspase-8 (CASP-8), we disclosed that CASP-8 regulates CRBN cleavage in cell lines. Site mapping experiments revealed that CRBN is cleaved after Asp9 upon CASP-8 activation, resulting in the reduced stability. Using myeloma as a model system, we further revealed that either inhibition or genetic depletion of CASP-8 enhances the anti-myeloma activity of lenalidomide (Len) by impairing CRBN cleavage, leading to the attenuated IKZF1 and IKZF3 protein levels and the reduced viability of myeloma cell lines and primary myeloma cells from patients. The present study discovered that the stability of the substrate receptor of an E3 ligase can be modulated by CASP-8 and suggested that administration of CASP-8 inhibitors enhances the overall effectiveness of Len-based combination therapy in myeloma.
Loss of G-Protein Pathway Suppressor 2 Promotes Tumor Growth Through Activation of AKT Signaling
Authors: Chan Stefanie , Smith Emma , Gao Yuan , Kwan Julian , Blum Benjamin C. , Tilston-Lunel Andrew M. , Turcinovic Isabella , Varelas Xaralabos , Cardamone Maria Dafne , Monti Stefano , Emili Andrew , Perissi Valentina
Published: January 2021

G Protein Suppressor 2 (GPS2) is a multifunctional protein that exerts important roles in inflammation and metabolism in adipose, liver, and immune cells. GPS2 has recently been identified as a significantly mutated gene in breast cancer and other malignancies and proposed to work as a putative tumor suppressor. However, molecular mechanisms by which GPS2 prevents cancer development and/or progression are largely unknown. Here, we have profiled the phenotypic changes induced by GPS2 depletion in MDA-MB-231 triple negative breast cancer cells and investigated the underlying molecular mechanisms. We found that GPS2-deleted MDA-MB-231 cells exhibited increased proliferative, migratory, and invasive propertiesin vitro, and conferred greater tumor burdenin vivoin an orthotopic xenograft mouse model. Transcriptomic, proteomic and phospho-proteomic profiling of GPS2-deleted MBA-MB-231 revealed a network of altered signals that relate to cell growth and PI3K/AKT signaling. Overlay of GPS2-regulated gene expression with MDA-MB-231 cells modified to express constitutively active AKT showed significant overlap, suggesting that sustained AKT activation is associated with loss of GPS2. Accordingly, we demonstrate that the pro-oncogenic phenotypes associated with GPS2 deletion are rescued by pharmacological inhibition of AKT with MK2206. Collectively, these observations confirm a tumor suppressor role for GPS2 and reveal that loss of GPS2 promotes breast cancer cell proliferation and tumor growth through uncontrolled activation of AKT signaling. Moreover, our study points to GPS2 as a potential biomarker for a subclass of breast cancers that would be responsive to PI3K-class inhibitor drugs.
Relationship Between Mitochondrial Structure and Bioenergetics in Pseudoxanthoma elasticum Dermal Fibroblasts
Authors: Lofaro Francesco Demetrio , Boraldi Federica , Garcia-Fernandez Maria , Estrella Lara , Valdivielso Pedro , Quaglino Daniela
Published: January 2021

Pseudoxanthoma elasticum (PXE) is a genetic disease considered as a paradigm of ectopic mineralization disorders, being characterized by multisystem clinical manifestations due to progressive calcification of skin, eyes, and the cardiovascular system, resembling an age-related phenotype. Although fibroblasts do not express the pathogenic ABCC6 gene, nevertheless these cells are still under investigation because they regulate connective tissue homeostasis, generating the “arena” where cells and extracellular matrix components can promote pathologic calcification and where activation of pro-osteogenic factors can be associated to pathways involving mitochondrial metabolism. The aim of the present study was to integrate structural and bioenergenetic features to deeply investigate mitochondria from control and from PXE fibroblasts cultured in standard conditions and to explore the role of mitochondria in the development of the PXE fibroblasts’ pathologic phenotype. Proteomic, biochemical, and morphological data provide new evidence that in basal culture conditions (1) the protein profile of PXE mitochondria reveals a number of differentially expressed proteins, suggesting changes in redox balance, oxidative phosphorylation, and calcium homeostasis in addition to modified structure and organization, (2) measure of oxygen consumption indicates that the PXE mitochondria have a low ability to cope with a sudden increased need for ATP via oxidative phosphorylation, (3) mitochondrial membranes are highly polarized in PXE fibroblasts, and this condition contributes to increased reactive oxygen species levels, (4) ultrastructural alterations in PXE mitochondria are associated with functional changes, and (5) PXE fibroblasts exhibit a more abundant, branched, and interconnected mitochondrial network compared to control cells, indicating that fusion prevail over fission events. In summary, the present study demonstrates that mitochondria are modified in PXE fibroblasts. Since mitochondria are key players in the development of the aging process, fibroblasts cultured from aged individuals or aged in vitro are more prone to calcify, and in PXE, calcified tissues remind features of premature aging syndromes; it can be hypothesized that mitochondria represent a common link contributing to the development of ectopic calcification in aging and in diseases. Therefore, ameliorating mitochondrial functions and cell metabolism could open new strategies to positively regulate a number of signaling pathways associated to pathologic calcification.
Identification of lncRNA-mRNA Regulatory Module to Explore the Pathogenesis and Prognosis of Melanoma
Authors: Zhang Jiaqi , Liu Hui , Zhang Wenhao , Li Yinfang , Fan Zhigang , Jiang Hua , Luo Judong
Published: January 2021

Skin cutaneous melanoma (SKCM) is an aggressive form of skin cancer that results in high mortality rate worldwide. It is vital to discover effective prognostic biomarkers and therapeutic targets for the treatment of melanoma. Long non-coding RNA (lncRNA) has been verified to play an essential role in the regulation of gene expression in diseases and tumors. Therefore, it is significant to explore the function of lncRNAs in the development and progression of SKCM. In this paper, a set of differentially expressed lncRNAs (DElncRNAs) and mRNAs (DEmRNAs) were first screened out using 471 cutaneous melanoma samples and 813 normal skin samples. Gene Ontology and KEGG pathway enrichment analysis were performed to obtain the significant function annotations and pathways of DEmRNAs. We also ran survival analysis on both DElncRNAs and DEmRNAs to identify prognostic-related lncRNAs and mRNAs. Next, a set of hub genes derived from protein-protein interaction (PPI) network analysis and lncRNA target genes screened from starbase-ENCORI database were integrated to construct a lncRNA-mRNA regulatory module, which includes 6 lncRNAs 4 target mRNAs. We further checked the capacity of these lncRNA and mRNA in the diagnosis of melanoma, and found that single lncRNA can effectively distinguish tumor and normal tissue. Moreover, we ran CMap analysis to select a list of small molecule drugs for SKCM, such as EGFR inhibitor AG-490, growth factor receptor inhibitor GW-441756 and apoptosis stimulant betulinic-acid, which have shown therapeutic effect in the treatment of melanoma.
Get Out and Stay Out: New Insights Into DNA Methylation Reprogramming in Mammals
Authors: Greenberg Maxim V. C.
Published: January 2021

Vertebrate genomes are marked by notably high levels of 5-cytosine DNA methylation (5meC). The clearest function of DNA methylation among members of the subphylum is repression of potentially deleterious transposable elements (TEs). However, enrichment in the bodies of protein coding genes and pericentromeric heterochromatin indicate an important role for 5meC in those genomic compartments as well. Moreover, DNA methylation plays an important role in silencing of germline-specific genes. Impaired function of major components of DNA methylation machinery results in lethality in fish, amphibians and mammals. Despite such apparent importance, mammals exhibit a dramatic loss and regain of DNA methylation in early embryogenesis prior to implantation, and then again in the cells specified for the germline. In this minireview we will highlight recent studies that shine light on two major aspects of embryonic DNA methylation reprogramming: (1) The mechanism of DNA methylation loss after fertilization and (2) the protection of discrete loci from ectopic DNA methylation deposition during reestablishment. Finally, we will conclude with some extrapolations for the evolutionary underpinnings of such extraordinary events that seemingly put the genome under unnecessary risk during a particularly vulnerable window of development.
The Viral Janus: Viruses as Aetiological Agents and Treatment Options in Colorectal Cancer
Authors: Turkington Christopher J. R. , Varadan Ambarish C. , Grenier Shea F. , Grasis Juris A.
Published: January 2021

In recent years, our understanding of the importance of microorganisms on and within our bodies has been revolutionized by the ability to characterize entire microbial communities. No more so is this true than in cases of disease. Community studies have revealed strong associations between microbial populations and disease states where such concomitance was previously absent from aetiology: including in cancers. The study of viruses, in particular, has benefited from the development of new community profiling techniques and we are now realising that their prominence within our physiology is nearly as broad as the diversity of the organisms themselves. Here, we examine the relationship between viruses and colorectal cancer (CRC), the leading cause of gastrointestinal cancer-related death worldwide. In CRC, viruses have been suggested to be involved in oncogenesis both directly, through infection of our cells, and indirectly, through modulating the composition of bacterial communities. Interestingly though, these characteristics have also led to their examination from another perspective—as options for treatment. Advances in our understanding of molecular and viral biology have caused many to look at viruses as potential modular biotherapeutics, where deleterious characteristics can be tamed and desirable characteristics exploited. In this article, we will explore both of these perspectives, covering how viral infections and involvement in microbiome dynamics may contribute to CRC, and examine ways in which viruses themselves could be harnessed to treat the very condition their contemporaries may have had a hand in creating.
Detection of Left Atrial Enlargement Using a Convolutional Neural Network-Enabled Electrocardiogram
Authors: Jiang Junrong , Deng Hai , Xue Yumei , Liao Hongtao , Wu Shulin
Published: January 2021

Background: Left atrial enlargement (LAE) can independently predict the development of a variety of cardiovascular diseases.

Objectives: This study sought to develop an artificial intelligence approach for the detection of LAE based on 12-lead electrocardiography (ECG).

Methods: The study population came from an epidemiological survey of heart disease in Guangzhou. Elderly people (3,391) over 65 years old who had both 10-s 12 lead ECG and echocardiography were enrolled in this study. The left atrial (LA) anteroposterior diameter >40 mm on echocardiography was diagnosed as LAE, and the LA anteroposterior diameter was indexed by body surface area (BSA) to classify LAE into different degrees. A convolutional neural network (CNN) was trained and validated to detect LAE from normal ECGs. The performance of the model was evaluated by calculating the area under the curve (AUC), accuracy, sensitivity, specificity, and F1 score.

Results: In this study, gender, obesity, hypertension, and valvular heart disease seemed to be related to left atrial enlargement. The AI-enabled ECG identified LAE with an AUC of 0.949 (95% CI: 0.911–0.987). The sensitivity, specificity, accuracy, precision, and F1 score were 84.0%, 92.0%, 88.0%, 91.3%, and 0.875, respectively. Physicians identified LAE with sensitivity, specificity, accuracy, precision, and F1 scores of 38.0%, 84.0%, 61.0%, 70.4%, and 0.494, respectively. In classifying LAE in different degrees, the AUCs of identifying normal, mild LAE, and moderate-severe LAE ECGs were 0.942 (95% CI: 0.903–0.981), 0.951 (95% CI: 0.917–0.987), and 0.998 (95% CI: 0.996–1.00), respectively. The sensitivity, specificity, accuracy, positive predictive value, and F1 scores of diagnosing mild LAE were 82.0%, 92.0%, 88.7%, 89.1%, and 0.854, while the sensitivity, specificity, accuracy, positive predictive value, and F1 scores of diagnosing moderate-severe LAE were 98.0%, 84.0%, 88.7%, 96.1%, and 0.969, respectively.

Conclusions: An AI-enabled ECG acquired during sinus rhythm permits identification of individuals with a high likelihood of LAE. This model requires further refinement and external validation, but it may hold promise for LAE screening.
The Ubiquitin Proteasome System in Periodontal Disease: A Comprehensive Review
Authors: Machado Vanessa , Carvalho Rui , Mendes José João , Botelho João
Published: January 2021

The turnover of intracellular proteins is a highly selective and regulated process. This process is responsible for avoiding injury and irreparable breakdown of cellular constituents. Its impairment disrupts cellular stability, integrity, and homeostasis. The ubiquitin-proteasome system (UPS) is responsible for this programmed degradation of most intracellular proteins. This process involves a cascade of enzymes that involves the ubiquitin conjugation to a target substrate protein, its recognition and degradation by the proteasome. The turn-over of intracellular proteins is a non-stop ubiquitous process that regulates a series of mechanisms, for instance transcription, translation, endocytosis. In addition, proteasome act by releasing peptides that may serve to other purposes, such as antigen presentation in immune actions and enzymatic flagging toward biosynthesis and gluconeogenesis. The role of the UPS impairment in periodontal diseases is gaining growing. This acquaintance might contribute to the development of novel therapeutic applications. Thus, this review focuses on the latest progresses on the role of the UPS and its signaling pathways in Periodontal Medicine. Furthermore, we discuss the potential of UPS-based drugs development to be used in periodontal disease therapy.
A Theoretical Framework on Reflection in Service Learning: Deepening Reflection Through Identity Development
Authors: Kawai Toru
Published: January 2021

In higher education, well-designed service learning combines service activities and academic knowledge in reflection, generating essential learning outcomes: academic enhancement, personal growth, and civic engagement. As research on reflection in service learning has shown, the process of reflection deepens through description of service experiences, examination of those experiences and articulation of learning. This article provides a theoretical explanation of deepening the reflection process by incorporating reflection theory and identity theory of college student development, professional development, and general identity development. Expanding the theoretical explanation of the reflective process clarifies the conditions of the deepening student reflection process in service learning in the following ways. First, it focuses on concrete experience then-and-there at that moment rather than abstract impressions by paying attention to personal dissonance in the experience. In addition, it finds discrepancies from differences of views, perspectives, and backgrounds between those of students and others. It connects outward exploration of those differences and inward exploration to construct internal voices toward self-authorship. The deep reflection process requires confronting contradictions through dialogical interplays among the I-positions of their own and others. It bridges discontinuities between past, present, and future selves by expanding the time perspective retrospectively and prospectively, and solving contradictions embedding in their prejudice. Furthermore, it activates plurality in social norms and values. The above conditions should be design principles for deepening critical and dialogical reflection in high-impact service learning. Through deepening reflection in service learning, it can be expected to activate mutuality and support generativity toward solidarity against hostility.
The Adrenal Cortex, an Underestimated Site of SARS-CoV-2 Infection
Authors: Mao Yanfei , Xu Bo , Guan Wenbin , Xu Dunfeng , Li Feng , Ren Rongrong , Zhu Xiaoyan , Gao Yuan , Jiang Lai
Published: January 2021

Background

The majority of the critically ill patients may have critical illness-related corticosteroid insufficiency (CIRCI). The therapeutic effect of dexamethasone may be related to its ability to improve cortical function. Recent study showed that dexamethasone can reduce COVID-19 deaths by up to one third in critically ill patients. The aim of this article is to investigate whether SARS-CoV-2 can attack the adrenal cortex to aggravate the relative adrenal insufficiency.

Methods

We summarized the clinical features of COVID-19 reported in currently available observational studies. ACE2 and TMPRSS2 expression was examined in human adrenal glands by immunohistochemical staining. We retrospectively analyzed serum cortisol levels in critically ill patients with or without COVID-19.

Results

High percentage of critically ill patients with SARS-COV-2 infection in the study were treated with vasopressors. ACE2 receptor and TMPRSS2 serine protease were colocalized in adrenocortical cells in zona fasciculata and zona reticularis. We collected plasma cortisol concentrations in nine critically ill patients with COVID-19. The cortisol levels of critically ill patients with COVID-19 were lower than those in non-COVID-19 critically ill group. Six of the nine COVID-19 critically ill patients had random plasma cortisol concentrations below 10 µg/dl, which met the criteria for the diagnosis of CIRCI.

Conclusion

We demonstrate that ACE2 and TMPRSS2 are colocalized in adrenocortical cells, and that the cortisol levels are lower in critically ill patients with COVID-19 as compared to those of non-COVID-19 critically ill patients. Based on our findings, we recommend measuring plasma cortisol level to guide hormonal therapy.
Identification of mRNAs and lncRNAs Involved in the Regulation of Follicle Development in Goat
Authors: Zhao Zhifeng , Zou Xian , Lu Tingting , Deng Ming , Li Yaokun , Guo Yongqing , Sun Baoli , Liu Guangbin , Liu Dewu
Published: January 2021

Follicular development and maturation has a significant impact on goat reproductive performance, and it is therefore important to understand the molecular basis of this process. The importance of long non-coding RNAs (lncRNAs) in mammalian reproduction has been established, but little is known about the roles of lncRNAs in different follicular stages, especially in goats. In this study, RNA sequencing (RNA-seq) of large follicles (>10 mm) and small follicles (<3 mm) of Chuanzhong black goats was performed to investigate the regulatory mechanisms of lncRNAs and mRNAs in follicular development and maturation. A total of 8 differentially expressed lncRNAs (DElncRNAs) and 1,799 DEmRNAs were identified, and the majority of these were upregulated in small follicles. MRO, TC2N, CDO1, and NTRK1 were potentially associated with follicular maturation. KEGG pathway analysis showed that the DEmRNAs involved in ovarian steroidogenesis (BMP6, CYP11A1, CYP19A1, 3BHSD, STAR, LHCGR, and CYP51A1) and cAMP signaling play roles in regulating follicular maturation and developmental inhibition respectively. Five target pairs of DElncRNA-DEmRNA, namely, ENSCHIT00000001255-OTX2, ENSCHIT00000006005-PEG3, ENSCHIT00000009455-PIWIL3, ENSCHIT00000007977-POMP, and ENSCHIT00000000834-ACTR3 in co-expression analysis provide a clue in follicular development and maturation of lncRNA-mRNA interaction. Our findings provide a valuable resource for lncRNA studies, and could potentially provide a deeper understanding of the genetic basis and molecular mechanisms of goat follicular development and maturation.
Intranasal Delivery of Inactivated Influenza Virus and Poly(I:C) Adsorbed Corn-Based Nanoparticle Vaccine Elicited Robust Antigen-Specific Cell-Mediated Immune Responses in Maternal Antibody Positive Nursery Pigs
Authors: Patil Veerupaxagouda , Renu Sankar , Feliciano-Ruiz Ninoshkaly , Han Yi , Ramesh Anikethana , Schrock Jennifer , Dhakal Santosh , HogenEsch Harm , Renukaradhya Gourapura J.
Published: January 2021

We designed the killed swine influenza A virus (SwIAV) H1N2 antigen (KAg) with polyriboinosinic:polyribocytidylic acid [(Poly(I:C)] adsorbed corn-derived Nano-11 particle based nanovaccine called Nano-11-KAg+Poly(I:C), and evaluated its immune correlates in maternally derived antibody (MDA)-positive pigs against a heterologous H1N1 SwIAV infection. Immunologically, in tracheobronchial lymph nodes (TBLN) detected enhanced H1N2-specific cytotoxic T-lymphocytes (CTLs) in Nano-11-KAg+Poly(I:C) vaccinates, and in commercial vaccinates detected CTLs with mainly IL-17A+ and early effector phenotypes specific to both H1N2 and H1N1 SwAIV. In commercial vaccinates, activated H1N2- and H1N1-specific IFNγ+&TNFα+, IL-17A+ and central memory T-helper/Memory cells, and in Nano-11-KAg+Poly(I:C) vaccinates H1N2-specific central memory, IFNγ+ and IFNγ+&TNFα+, and H1N1-specific IL-17A+ T-helper/Memory cells were observed. Systemically, Nano-11-KAg+Poly(I:C) vaccine augmented H1N2-specific IFNγ+ CTLs and H1N1-specific IFNγ+ T-helper/Memory cells, and commercial vaccine boosted H1N2- specific early effector CTLs and H1N1-specific IFNγ+&TNFα+ CTLs, as well as H1N2- and H1N1-specific T-helper/Memory cells with central memory, IFNγ+&TNFα+, and IL-17A+ phenotypes. Remarkably, commercial vaccine induced an increase in H1N1-specific T-helper cells in TBLN and naive T-helper cells in both TBLN and peripheral blood mononuclear cells (PBMCs), while H1N1- and H1N2-specific only T-helper cells were augmented in Nano-11-KAg+Poly(I:C) vaccinates in both TBLN and PBMCs. Furthermore, the Nano-11-KAg+Poly(I:C) vaccine stimulated robust cross-reactive IgG and secretory IgA (SIgA) responses in lungs, while the commercial vaccine elicited high levels of serum and lung IgG and serum hemagglutination inhibition (HI) titers. In conclusion, despite vast genetic difference (77% in HA gene identity) between the vaccine H1N2 and H1N1 challenge viruses in Nano-11-KAg+Poly(I:C) vaccinates, compared to over 95% identity between H1N1 of commercial vaccine and challenge viruses, the virus load and macroscopic lesions in the lungs of both types of vaccinates were comparable, but the Nano-11-KAg+Poly(I:C) vaccine cleared the virus from the nasal passage better. These data suggested the important role played by Nano-11 and Poly(I:C) in the induction of polyfunctional, cross-protective cell-mediated immunity against SwIAV in MDA-positive pigs.
Actin Remodeling Defects Leading to Autoinflammation and Immune Dysregulation
Authors: Papa Riccardo , Penco Federica , Volpi Stefano , Gattorno Marco
Published: January 2021

A growing number of monogenic immune-mediated diseases have been related to genes involved in pathways of actin cytoskeleton remodeling. Increasing evidences associate cytoskeleton defects to autoinflammatory diseases and primary immunodeficiencies. We reviewed the pathways of actin cytoskeleton remodeling in order to identify inflammatory and immunological manifestations associated to pathological variants. We list more than twenty monogenic diseases, ranging from pure autoinflammatory conditions as familial Mediterranean fever, mevalonate kinase deficiency and PAPA syndrome, to classic and novel primary immunodeficiencies as Wiskott-Aldrich syndrome and DOCK8 deficiency, characterized by the presence of concomitant inflammatory and autoimmune manifestations, such as vasculitis and cytopenia, to severe and recurrent infections. We classify these disorders according to the role of the mutant gene in actin cytoskeleton remodeling, and in particular as disorders of transcription, elongation, branching and activation of actin. This expanding field of rare immune disorders offers a new perspective to all immunologists to better understand the physiological and pathological role of actin cytoskeleton in cells of innate and adaptive immunity.
Protocol for Sampling Sequential Fin Spine Growth Intervals for Isotope Analysis in the Atlantic Bluefin Tuna
Authors: Luque Patricia Lastra , Sakai Saburo , Murua Hilario , Arrizabalaga Haritz
Published: January 2021

Background

Micromilled fish otoliths (ear bones) have been widely used for stable oxygen (δ18O) and carbon (δ13C) isotope analysis. The first dorsal fin spine is routinely used for ageing in the eastern Atlantic bluefin tuna (ABFT) population; however, stable isotope analysis remains unapplied in this hard structure. The objective of the present protocol is to achieve a sequential sampling of growth layers in the ABFT fin spine at high spatial resolution and along growth trajectory for stable isotope analysis.

Methods

We used a micromilling and micro-powder collecting technique for sequentially sampling annual growth layers of the fin spine bone collected from freshly caught ABFT. We assessed the carbonate content and the optimal drilling amount to ensure enough powder quantity was recovered from each annual growth band to accommodate accurate measurement of the δ18O and δ13C values.

Results

The optimal drilling path included 20 drilling lines in 49 μm, with 400 μm depth and 900 μm-line width, which represents a time resolution of about 2.5 months. The minimum powder quantity required from each annual growth layer for δ18O and δ13C isotope analysis was approximately 180 μg because the carbonate contents in the fin spine powder was 5–6%. The δ18O profile assayed in the last growth intervals coincide with the surface δ18Oseawater for the southern west waters off the Norwegian coast, where the tuna was caught. The fluctuating pattern in the second growth interval may reflect certain fidelity to the Norwegian SE and Swedish SW coast and/or otherwise to the US west coast. The δ13Cspine displayed a larger temporal variability along the growth transect that may be explained by several factors including differences in habitat use, behavior, and even trophic level.

Conclusion

Based on the result obtained, we present an optimal standard protocol for the sampling of sequential, annually formed growth layers in the fin spine bone for stable isotope analysis using the micro-milling as a high precision technique. This protocol is particularly useful in endangered and/or protected species for which fin spines represent a non-lethal alternative to otoliths opening new research avenues to improve their management and conservation.