bluuebunny/crossref_metadata_2025_split huggingface dataset:
Northern blotting of endogenous full-length human-specific LINE-1 RNA
Authors: Alkailani Maisa I 
Published: May 2024

Abstract
LINE-1 belongs to a family of DNA elements that move to new locations in the genome in a process called “retrotransposition.” This is achieved by a copy-and-paste mechanism with the aid of an RNA intermediate. The full-length LINE-1 is responsible for most retrotransposition activity in the human genome. Detecting the active LINE-1 RNA at the endogenous level is challenging due to its small percentage among inactive copies and its different forms of transcripts. Here, we describe a method of designing RNA probes to detect active LINE-1 by northern blotting and use optimized conditions and tools to make the detection practical. This method uses a classical long RNA probe and provides an alternative way to detect LINE-1 RNA using multiple short RNA probes.
Implications of lung shunt fraction calculation discrepancy in Yttrium-90 radioembolization treatment from 2D planar vs 3D single photon emission CT imaging
Authors: Gamero Kubota Paula , Molvar Christopher , Wagner Robert , Allam Emad , Halama James , James Judy R 
Published: May 2024

Abstract
The safety and efficacy of Yttrium-90 (Y-90) radio-embolization therapy is partly dependent on the lung shunt fraction (LSF). There may be a notable disparity between LSF when calculated using 2D planar imaging vs 3D single photon emission CT (SPECT); this can affect the total allowable Y-90 dose delivered and therefore change the effectiveness of the procedure. The case presented demonstrates an 81% decrease in LSF when calculated by SPECT as compared to 2D planar imaging. This case highlights the importance of considering the imaging technique and the potential discrepancies that can arise between planar and SPECT imaging in LSF assessment.
Chelate extraction of metal ions in aqueous/chloroform system based on molecular crowding environment
Authors: Miyagawa Akihisa , Komatsu Hiroyuki , Nagatomo Shigenori , Nakatani Kiyoharu 
Published: May 2024

Abstract
In this study, we propose a novel concept for the solvent extraction of metal ions (Co, Zn, and Pb) by mimicking a molecular crowding environment using dextran (Dex). The metal ions were extracted from the aqueous phase into the organic phase (chloroform) in the presence of 8-hydroxyquinoline (HQ). The extraction constant of the metal complex (Kex) increased with increasing Dex concentration (CDex) for all metal ions. When examining the dependence of CDex on the four equilibrium constants (distribution coefficient of HQ, acid dissociation of HQ, complexation constant of metal complex (β), and distribution coefficient of the metal complex) that contribute to Kex, only β increased with CDex. This suggests that an increase in, β, a parameter reflecting the molecular crowding effect, results in an increase in Kex. The increase in β was analyzed based on volume exclusion and osmotic pressure effects. The analytical model effectively explained the enhanced the complexation due to the increase in β and volume exclusion, whereas the osmotic pressure suppressed β. Consequently, we unveiled the effect of molecular crowding on the solvent extraction of metal ions for the first time.
Soberanas de nuestros cuerpos: networked framing of abortion legalization in Argentina
Authors: Cabas-Mijares Ayleen , Wichowsky Amber 
Published: May 2024

Abstract
This study presents a critical discourse analysis (CDA) of the networked frames built in Twitter trending hashtags by proponents and opponents of abortion legalization in Argentina. The online discourses exemplified the dynamics of networked framing, where activists reshaped their framing of issues through crowdsourcing practices. While both sides focused their attention on defining violence and deservingness, the impact of abortion on democracy, and the appropriate geopolitical scope of the debate, they developed stark oppositional views. Supporters of legalization defined illegal abortion as state-sponsored violence against gestating people and highlighted transnational solidarities. On the other hand, pro-life users identified legal abortion as state-sponsored violence against the unborn and accused pro-abortion politicians of subverting the democratic rights of the pro-life majority. Both movements portrayed themselves as embodying reason and righteousness while projecting negative connotations onto their counterparts. Our findings provide a nuanced examination of the discursive work of Global South feminists whose advocacy shifted policy against formidable opposition.
Heat exposure limits pentose phosphate pathway activity in bumblebees
Authors: Rossi Giulia S , Elbassiouny Alaa , Jamison Jerrica , Welch Jr. Kenneth C 
Published: May 2024

Abstract
Bumblebee populations across the globe are experiencing substantial declines due to climate change, with major consequences for pollination services in both natural and agricultural settings. Using an economically important species, Bombus impatiens, we explored the physiological mechanisms that may cause susceptibility to extreme heat events. We tested the hypothesis that heat exposure limits the activity of the pentose phosphate pathway (PPP)—a parallel pathway to glycolysis that can use nectar sugar to generate antioxidant potential and combat oxidative stress. Using isotopically labelled glucose, we tracked PPP activity in B. impatiens at rest, during exercise and during a post-exercise recovery period under two different temperature regimes (22°C and 32°C). We found that the PPP is routinely used by B. impatiens at moderate temperatures, but that its activity is markedly reduced when ATP demands are high, such as during periods of exercise and heat exposure. We also exposed B. impatiens to either 22°C or 32°C for 5 hours and assessed levels of oxidative damage (lipid peroxidation, protein carbonyls) and antioxidant potential [reduced (GSH) and oxidized (GSSG) glutathione concentrations]. Interestingly, bees exhibited little oxidative damage after the thermal exposure, but we found a lower GSH:GSSG ratio in 32°C-exposed bees, reflecting lower antioxidant potential. Overall, our study demonstrates that acute heat stress severely limits PPP activity and may constrain antioxidant potential in B. impatiens. The repeated attenuation of this pathway in a warming climate may have more severe physiological consequences for this species, with potential implications for pollination services across North America.
miR-1 modulates HDAC4, calmodulin and MAPK1/Erk2 during early atrial differentiation
Authors: Franco D , Garcia-Padilla C , Lozano-Velasco E , Garcia-Lopez V , Aranega A , Garcia-Martinez V , Lopez-Sanchez C 
Published: May 2024

Abstract

Funding Acknowledgements
None.

Introduction
A large diversity of epigenetic factors, such as microRNAs and histones modifications, are known to be capable of regulating gene expression without altering DNA sequence itself. In particular, miR-1 is considered the first essential microRNA in cardiac development.

Purpose
The aim of this work is to analyze the miR-1 modulation role in cardiac chamber differentiation through specific signaling pathways.

Methods
By means of microinjections in both primitive endocardial tubes of chick embryo culture, gain- and loss-of-function experiments were performed with premiR-1 and anti-miR-1, respectively. Subsequently, embryos were subjected to whole mount in situ hybridization with Tbx5, Gata4 and AMHC1 probes, as well as immunohistochemistry with Mef2c, HDAC4, Calmodulin/Calm1 and MAPK1/Erk2 antibodies. Also, we carried out RT-qPCR analysis of control and experimental embryos, including CRABPI, CRABPII and retinoic acid receptors (RAR and RXR).

Results
Our results reveal that miR-1 increases specific atrial gene expression such as Tbx5, Gata4 and AMHC1, while this microRNA diminishes Mef2c expression. Furthermore, we observed that miR-1 upregulates CRABPII and RARß, and downregulates CRABPI, which are three crucial factors in retinoic acid signaling pathway. Interestingly, we also observed that miR-1 actively interacts with HDAC4, Calmodulin and Erk2/MAPK1, key factors involved in Mef2c regulation.

Conclusion
All these data suggest that miR-1 functions as an epigenetic factor modulating complementary actions performed by retinoic acid and Mef2c, which are required to properly assign cells as sinoatrial precursors.
MicroRNA-127-3p regulates inflammatory response in injury-induced vascular remodeling and cellular senescence in vascular smooth muscle cells through mTor
Authors: Daniel F , Kalies K , Hehl L , Guerlach S , Koester J M , Boettcher S , Knoepp K , Dutzmann J , Sedding D 
Published: May 2024

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Martin-Luther-Universität Halle-Wittenberg

Background and Purpose
The accumulation of senescent cells in the vasculature contributes to impaired vascular function over the lifetime. Micro RNAs (miRs) are promising therapeutic targets due to their cell-specific functions. Targeting miR-127-3p emerges as an attractive approach to enhance vascular healing and limit angioplasty-related complications.
Here, we focus on elucidating the role of miR-127-3p in vascular smooth muscle cells (VSMC) to characterize their role in cellular function and vascular senescence.

Methods
Utilizing reverse transcription and qRT-PCR we checked expression levels of miR-127-3p in human and murine aortic VSMCs. After overexpression and downregulation via transfection of pre-miR and antagomiR, we evaluated the influence of miR-127-3p on cellular functions such as proliferation and migration of human (non-) replicative senescent VSMCs. Potential targets were selected via target prediction analysis and verified on mRNA and protein levels. Morphological changes were visualized via high-resolution fluorescence microscopy. Senescence marker expression was checked after transfection by qRT-PCR.

Results
MiR-127-3p is highly upregulated in femoral artery tissue on day 10 after wire-induced injury in C57Bl6 mice, serving as restenosis model (p&lt;0.001) and aged C57BL/6 mice (3 months vs. 22 months, p&lt;0.0001). Replicative senescent human VSMC exhibit increased miR-127-3p expression (p&lt;0.05). In vitro, miR-127-3p overexpression reduces proliferation in senescent (p&lt;0.001) and non-senescent (p&lt;0.01) cells. It hinders migration in non-senescent cells (p&lt;0.0001) but has a reverse effect in senescent cells (p&lt;0.01). Potential targets affecting cellular functions, inflammatory pathways, and cellular aging were identified as mTOR, AXL, and TEAD3 (p&lt;0.05). Transfection of pre-miR leads to increase in cell size and the acquisition of a flattened morphology of human VSMC. P14arf and laminB1, as senescence-associated genes, showed downregulation after overexpression of miR-127-3p on mRNA level (p&lt;0.05). When VSMCs were stimulated with anti-miR-127-3p, increased mRNA levels of p14arf and laminB1 were detected (p&lt;0.05).

Conclusion
This study identifies miR-127-3p as a contributor to cellular functions including cellular proliferation and migration and seems to have opposing effects in non- and replicative senescent VSMC. Further research is needed to validate miR-127-3p as a therapeutic target for vascular aging and remodeling in vitro and in vivo.
Acquired control over proliferation and differentiation of ventricular cardiomyocytes to break down the mechanisms of cardiac repair
Authors: Ramkisoensing A , Zhang J , Pijnappels D A , De Vries A A F 
Published: May 2024

Abstract
Funding Acknowledgements
None.
Loss of myocardial tissue remains a leading cause of morbidity and mortality by causing heart rhythm disturbances and heart failure. The (pre-)clinical effects of inducing cardiac regeneration by cardiac cell therapy have been disappointing. In order to find new curative options for these disabling cardiac diseases a shift of focus towards steering the heart to regenerate itself is needed. The aim of this study was to generate a robust ventricular cardiomyocyte model in which proliferation and differentiation could be tightly controlled to investigate cardiac regeneration. To accomplish this we have generated a line of immortalized neonatal rat ventricular cardiomyocytes (nrVMs) through expression of the cell cycle regulators cyclin-dependent kinase 4 and cyclin D1 (CDK4-D1). The CDK4-D1 expressing nrVMs were positive for the proliferation marker Ki67 in contrast to control nrVMs. Cardiomyogenic differentiation could still be induced after multiple passages (&gt; passage 10) as evidenced by immunocytochemistry and optical voltage mapping. These contractile and excitable cells were indeed positive for the cardiac-specific sarcomeric proteins alpha actinin and troponin T in a sarcomeric pattern similar to native nrVMs. Passaging of the control nrVM cultures, however, ultimately led to a senescent cell population lacking sarcomeric markers after passage 3. Electrophysiological analysis of immortalized monolayers, after inducing cardiomyogenic differentiation, showed not only uniform propagation of electrical signal throughout the cultures, but also re-entrant activity although at low frequency. In conclusion, these data show that were able to immortalize ventricular cardiomyocytes by preserving their contractile and excitable phenotype upon significant expansion. Further optimisation of this model will lead to a research model of ventricular cardiomyocytes which will give us the opportunity to unravel the process of cardiomyogenesis and uncover mechanisms which could allow us to enhance the ability of the heart to regenerate itself.
MiR-92b-3p modulates apoptosis in smooth muscle cells through BCL2L11 regulation, unravelling novel therapeutic strategies for unstable plaques
Authors: Koester J , Kalies K , Boettcher S , Guerlach S , Daniel F , Hehl L , Sedding D G 
Published: May 2024

Abstract
Funding Acknowledgements
None.
Smooth muscle cell (SMC) apoptosis is a contributory factor to plaque rupture in atherosclerosis, thereby promoting plaque destabilization through various mechanisms such as thrombogenic activation and plaque microcalcification. Immunohistochemical analysis has revealed elevated expression of proteins associated with intrinsic apoptosis, such as BCL2L11 (Bim), in unstable atherosclerotic plaques. Our hypothesis posits that Bim is a direct target of miR-92b-3p, an understudied microRNA, prompting an investigation into its role in regulating apoptosis in SMCs. Our objective is to discern a novel therapeutic axis for preventing plaque rupture by employing miRNA-based targeted interventions.
Within a model of primary human vascular cells, we employed various in vitro techniques, including PCR, Western Blot, Live cell imaging, and ELISA. Lipotransfection facilitated transient modulation of miR-92b-3p, using both knockdown and elevation, while an siRNA transfection model was also utilized.
MiR-92b-3p exhibits robust expression in late atherosclerotic lesions in vivo (p&lt;0.001) and demonstrates heightened levels in smooth muscle cells (SMCs) compared to endothelial cells (ECs) in vitro (p&lt;0.01) and in vivo in respective tissues (p&lt;0.01). Bim, a direct target of miR-92b-3p, experiences upregulation post-miR-92b-3p knockdown and downregulation following pre-miR-92b-3p introduction into SMCs, both at mRNA and protein levels (p&lt;0.01, p&lt;0.01). Apoptosis increases in growth-medium cultured SMCs after miR-92b-3p knockdown (p&lt;0.05), but not in senescent SMCs, ECs, or basal-medium treated SMCs. Live-cell imaging reveals a reduction in SMC count after anti-miR-92b-3p transfection (p&lt;0.01), accompanied by cell shrinkage indicative of apoptosis 72 hours post-transfection (p&lt;0.05). Growth factor stimulation results in miR-92b-3p upregulation (p&gt;0.05) and Bim downregulation in SMCs. SiRNA knockdown of BIM in SMCs abolishes apoptosis induction through miR-92b-3p knockdown (p&lt;0.05). Conversely, in apoptotic SMCs, miR-92b-3p upregulation protects cells from apoptosis (p&lt;0.05).
MiR-92b-3p effectively modulates mitochondrial apoptosis in SMCs by targeting BIM. This regulatory impact is exclusive to proliferative SMCs, known for their deleterious involvement in atherosclerotic plaques. Cumulatively, the elevation of miR-92b-3p via targeted miRNA therapeutics or miRNA-drug interactions in the advanced stages of atherosclerosis presents a potential therapeutic avenue for averting plaque rupture.
Mechanical load regulates the proliferation of multiple cell types in the heart
Authors: Ciucci G , Colliva A , Vodret S , Texler B , Cardini B , Oberhuber R , Vuerich R , Zago E , Maglione M , Sinagra G , Giacca M , Eschenhagen T , Golino P , Loffredo F , Zacchigna S 
Published: May 2024

Abstract

Funding Acknowledgements
None.

Introduction
The adult mammalian heart has a poor regenerative and angiogenic potential. At the same time, both primary cardiac tumors and cardiac metastases are extremely rare. This suggests that shared mechanisms may halt the proliferation of cardiac (both cardiomyocyte and endothelial) and cancer cells. Among the mechanisms that have been claimed to be responsible for the loss of proliferative potential of cardiomyocytes early after birth is a sudden increase in mechanical load.

Purpose
Here, we investigate whether alterations in mechanical load affect the proliferation of multiple cell types in the heart.

Methods
Adult heart unloading in vivo was achieved using a mouse model of heterotopic heart transplantation, followed by EdU injection to label proliferating cells. Engineered heart tissues (EHTs) were generated using neonatal cardiomyocytes and fibroblasts, with a modified protocol allowing to finely tune mechanical load. Primary cardiac endothelial cells and multiple types of cancer cells (lung, melanoma and colon cancer) along with adult cardiac endothelial cells were included in EHTs.

Results
In line with our hypothesis, unloaded, heterotransplanted hearts showed a significant number of proliferating cardiomyocytes and endothelial cells, which were almost absent in native hearts. While cancer cells did not grow in native hearts, they massively proliferated in unloaded hearts. Consistent with in vivo results, mechanical unloading in EHTs significantly increased the percentage of EdU-positive cardiomyocytes, endothelial and cancer cells. In contrast, increasing afterload reduced cell proliferation in all cell types. Mechanistically, we identified Nesprin2 as an essential factor in sensing mechanical load and translating it into a cell proliferation arrest.

Conclusions
Overall, these data indicate that variations in mechanical load have a dramatic effect on the proliferation of multiple cell types within the heart tissue, including cardiomyocytes, endothelial and cancer cells and that Nesprin2 is required to mediate this effect.Unloading in the heartUnloading in EHT
SGK1 inhibition normalizes action potential duration in transgenic LQT2 rabbits but not in LQT1, suggesting a novel gene-specific therapeutic approach in long QT syndrome
Authors: Barbieri M , Lourdaour J , Alerni N , Giammarino L , Nimani S , Matas L , Horvath A , Odening K E 
Published: May 2024

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): SNSF (Swiss National Science Fundation)

Background
Increasing evidence suggests that secondary remodeling in Na+ and Ca2+ homeostasis may further enhance the propensity to develop arrhythmias in K-channelopathies LQT1 and LQT2 syndrome. In this context, the Serum and Glucocorticoid regulated Kinase-1 (SGK-1) was recently identified as a major regulator of sodium channels in cardiomyocytes. It is mainly activated in pathological conditions, leading to increased INaLate, prolongation of action potential duration (APD) and increased susceptibility to ventricular arrhythmias, recapitulating LQTS phenotype. SGK1 inhibition might hence be a novel therapeutic target also in genetic LQTS subtypes.

Methods
In cardiomyocytes (CM) isolated from wild type (WT), LQT1 and LQT2 adult rabbits patch-clamp experiments were performed to record AP with SGK1-inhibitor(SGK1-inh, 3 µM) or DMSO vehicle control.

Results
In the perforated patch configuration at 37°C, 3 μM of SGK1-inh shortened APD90 in LQT2 CMs at 1 Hz (DMSO-control [ms]: 477.3 vs treated cells[ms]: 362.8 p &lt; 0.0001) and at 2 Hz (DMSO-cells: 325.7 vs treated cells:295.4,p &lt;0.05). In contrast, in WT and LQT1 CMs, no effect on APD90 was observed neither at 1 Hz (WT, DMSO-cells: 329.2 vs treated cells: 301.6 p=ns; LQT1, DMSO-cells: 429.8 vs treated cells: 431.6, p=ns) nor at 2 Hz (WT, DMSO-cells:256.8 vs treated cells:242 p=ns; LQT1,DMSO cells:310.4 vs treated cells: 315.8,p=ns). Importantly, the SGK1-inhibitor effect in LQT2 rabbit CMs was so pronounced that it normalized APD to the wildtype level (LQT2+SGK1-Inh: 362.8 vs WT:329.2, ps=ns). Furthermore, short-term-variability in APD was markedly reduced by SGK1-Inh in LQT2, indicating an antiarrhythmic effect.

Conclusions
SGK1-inhibition exerts a beneficial APD-shortening effect only in LQT2, indicating a gene-specific efficacy. The underlying mechanism is still unclear. We hypothesize that the SGK1 pathway/INaLate activation in LQT1 might not be as active as in LQT2. We are currently conducting more experiments on potential genotype differences in SGK1 activity and SGK1-mediated INaLate inhibition to further elucidate the differences.Effects of SGK1-inh on AP characteristic
The smooth-walled human RVOT contains trabeculations that cause conduction delay
Authors: Jensen B , Bosada F M , Blok M , Scholman K T , Efimov I R , Boukens B J 
Published: May 2024

Abstract

Funding Acknowledgements
None.

Background
The right ventricular outflow tract (RVOT) is the outlet from the right ventricle and is the initiating substrate of life-threatening arrhythmias. While the luminal wall of the RVOT is often assumed to be without the complex trabecular meshwork that characterizes the right ventricular free wall, the anatomy of the RVOT is an understudied subject.

Aim
to investigate whether trabeculations occur in the RVOT and to assess whether this impacts electrical propagation.

Methods
Human hearts were investigated using high-resolution MRI, histology, RNA-sequencing, and optical and electrical mapping.

Results
We used high-resolution MRI and serial sectioning to reconstruct the macroscopic details of the human RVOT and identified cases exhibiting much trabeculation. The smooth lumen of the RVOT varied between 9% and 23% of the total RV anterior surface (N=11). Histological analysis on additional six hearts indicated that the RVOT compact layer is thinner when trabeculations are present. RNA sequencing of four human donor hearts revealed enrichment in the subendocardial region of 88 genes associated with cardiac conduction and trabeculations (P adjusted&lt;0.05). Finally, we selected two human donor hearts showing trabeculations in the RVOT from which we generated wedge preparation and performed optical and electrical mapping. The trabecular regions demonstrated high degree of fractionation when compared to non-trabeculated regions, which coincided with delayed activation.

Conclusion
Trabeculations are found in the RVOT, and their extent varies among individuals. This impacts on the thickness of the compact wall in the RVOT, restricting the depth of tissue at which clinical interventions can be performed, as well as influencing electrical propagation and possible arrhythmogenicity.