bluuebunny/crossref_metadata_2025_split huggingface dataset:
Recombinant Human TSH Fails to Induce the Proliferation and Migration of Papillary Thyroid Carcinoma Cell Lines
Authors: Kalampounias Georgios , Varemmenou Athina , Aronis Christos , Mamali Irene , Shaukat Athanasios-Nasir , Chartoumpekis Dionysios V , Katsoris Panagiotis , Michalaki Marina 
Published: April 2024

AbstractThyrotropin (TSH) suppression is required in the management of patients with papillary thyroid carcinoma (PTC) to improve their outcomes, inevitably causing iatrogenic thyrotoxicosis. Nevertheless, the evidence supporting this practice remains limited and weak, andin vitrostudies examining the mitogenic effects of TSH in cancerous cells used supraphysiological doses of bovine TSH, which produced conflicting results. Our study explores for the first time the impact of human recombinant thyrotropin (rh-TSH) on human PTC cell lines (K1 and TPC-1) that were transformed to overexpress the thyrotropin receptor (TSHR). The cells were treated with escalating doses of rh-TSH under various conditions, such as the presence or absence of insulin. The expression levels ofTSHRandthyroglobulin(Tg) were determined, and subsequently, the proliferation and migration of both transformed and non-transformed cells were assessed. Under the conditions employed, rh-TSH was not adequate to induce either the proliferation or the migration rate of the cells, whileTgexpression was increased. Our experiments indicate that clinically relevant concentrations of rh-TSH cannot induce proliferation and migration in PTC cell lines, even after overexpression of TSHR. Further research is warranted to dissect the underlying molecular mechanisms, and these results could translate into better management of PTC patients.
Rat Anterior Insula Symmetrically Represents Tickling-Induced Playful Emotions
Authors: Dagher Sarah , Ishiyama Shimpei 
Published: April 2024

AbstractSocial play, an integral aspect of animal behavior, is inherently associated with positive emotions, yet the neuronal underpinnings of these playful states remain inadequately explored. We examined the anterior insula’s involvement in processing tickle-induced playful emotions in rats. Our findings revealed diverse patterns of insular activity during tickling, with 20% of the recorded units displaying strong activation, and another 20% exhibiting inhibition. These units responded similarly to other playful contexts, such as gentle touch and hand chase, but not to neutral locomotion. Tickle-activated units demonstrated a positive correlation of firing rates with appetitive vocalization rates, whereas tickle-inhibited units showed a negative correlation. Distinct spike waveforms were associated with the tickle response patterns, suggesting potential cell-type dependencies. However, pharmacological manipulation of the global anterior insula did not yield observable effects on play behavior in rats. Anterograde tracing revealed extensive insular projections to areas including the amygdala and nucleus accumbens. Taken together, our findings suggest that the anterior insula symmetrically represents tickle-induced playful emotional states.
Dynamic Tracking of Native Precursors in Adult Mice
Authors: Liu Suying , Adams Sarah E. , Zheng Haotian , Ehnot Juliana , Jung Seul K. , Jeffrey Greer , Menna Theresa , Purton Louise E. , Lee Hongzhe , Kurre Peter 
Published: April 2024

AbstractHematopoietic dysfunction has been associated with a reduction in the number of active precursors. However, precursor quantification at homeostasis and under diseased conditions is constrained by the scarcity of available methods. To address this issue, we optimized a method for quantifying a wide range of hematopoietic precursors. Assuming the random induction of a stable label in precursors following a binomial distribution, estimates depend on the inverse correlation between precursor numbers and the variance of precursor labeling among independent samples. Experimentally validated to cover the full dynamic range of hematopoietic precursors in mice (1 to 105), we utilized this approach to demonstrate that thousands of precursors, which emerge after modest expansion during fetal-to-adult transition, contribute to native and perturbed hematopoiesis. We further estimated the number of precursors in a mouse model of Fanconi Anemia, showcasing how repopulation deficits can be classified as autologous (cell proliferation) and non-autologous (lack of precursor). Our results support an accessible and reliable approach for precursor quantification, emphasizing the contemporary perspective that native hematopoiesis is highly polyclonal.
Biomaterial Scaffold Stiffness Influences the Foreign Body Reaction, Tissue Stiffness, Angiogenesis and Neuroregeneration in Spinal Cord Injury
Authors: Zheng Yifeng , Nützl Maximilian , Schackel Thomas , Chen Jing , Weidner Norbert , Müller Rainer , Puttagunta Radhika 
Published: April 2024

AbstractBiomaterial scaffold engineering presents great potential in promoting axonal regrowth after spinal cord injury (SCI), yet persistent challenges remain, including the surrounding host foreign body reaction and improper host-implant integration. Recent advances in mechanobiology spark interest in optimizing the mechanical properties of biomaterial scaffolds to alleviate the foreign body reaction and facilitate seamless integration. The impact of scaffold stiffness on injured spinal cords has not been thoroughly investigated. Herein, we introduce stiffness-varied alginate anisotropic capillary hydrogel scaffolds implanted into adult rat C5 spinal cords post-lateral hemisection. Four weeks post-implantation, scaffolds with a stiffness approaching that of the spinal cord effectively minimize the host foreign body reaction via yes-associated protein (YAP) nuclear translocation. Concurrently, the softest scaffolds maximize cell infiltration and angiogenesis, fostering significant axonal regrowth but limiting the rostral-caudal linear growth. Furthermore, as measured by atomic force microscopy (AFM), the surrounding spinal cord softens when in contact with the stiffest scaffold while maintaining a natural level in contact with the softest one. In conclusion, our findings underscore the pivotal role of stiffness in scaffold engineering for SCIin vivo, paving the way for the optimal development of efficacious biomaterial scaffolds for tissue engineering in the central nervous system.
Tiny amphibious insects use tripod gait for seamless transition across land, water, and duckweed
Authors: O’Neil Johnathan N. , Yung Kai Lauren , Difini Gaetano , Walker Holden , Bhamla M. Saad 
Published: April 2024

AbstractInsects exhibit remarkable adaptability in their locomotive strategies across diverse environments, a crucial trait for foraging, survival, and predator avoidance.Microvelia, tiny 2-3 mm insects that adeptly walk on water surfaces, exemplify this adaptability by using the alternating tripod gait in both aquatic and terrestrial terrains. These insects commonly inhabit low-flow ponds and streams cluttered with natural debris like leaves, twigs, and duckweed. Using high-speed imaging and pose-estimation software, we analyzeMicrovelia spp.’smovement across water, sandpaper (simulating land), and varying duckweed densities (10%, 25%, and 50% coverage). Our results revealMicroveliamaintain consistent joint angles and strides of their upper and hind legs across all duckweed coverages, mirroring those seen on sandpaper.Microveliaadjust the stride length of their middle legs based on the amount of duckweed present, decreasing with increased duckweed coverage and at 50% duckweed coverage, their middle legs’ strides closely mimic their strides on sandpaper. Notably,Microveliaachieve speeds up to 56 body lengths per second on water, nearly double those observed on sandpaper and duckweed (both rough, frictional surfaces), highlighting their higher speeds on low friction surfaces such as the water’s surface. This study highlightsMicrovelia’s ecological adaptability, setting the stage for advancements in amphibious robotics that emulate their unique tripod gait for navigating complex terrains.
DTX3L ubiquitin ligase ubiquitinates single-stranded nucleic acids
Authors: Dearlove Emily L. , Chatrin Chatrin , Buetow Lori , Ahmed Syed F. , Schmidt Tobias , Bushell Martin , Smith Brian O. , Huang Danny T. 
Published: April 2024

AbstractUbiquitination typically involves covalent linking of ubiquitin (Ub) to a lysine residue on a protein substrate. Recently, new facets of this process have emerged, including Ub modification of non-proteinaceous substrates like ADP-ribose by the DELTEX E3 ligase family. Here we show that the DELTEX family member DTX3L expands this non-proteinaceous substrate repertoire to include single-stranded DNA and RNA. Although the N-terminal region of DTX3L contains single-stranded nucleic acid binding domains and motifs, the minimal catalytically competent fragment comprises the C-terminal RING and DTC domains (RD). DTX3L-RD catalyses ubiquitination of the 3’-end of single-stranded DNA and RNA, as well as double-stranded DNA with a 3’ overhang of two or more nucleotides. This modification is reversibly cleaved by deubiquitinases. NMR and biochemical analyses reveal that the DTC domain binds single-stranded DNA and facilitates the catalysis of Ub transfer from RING-bound E2-conjugated Ub. Our study unveils the direct ubiquitination of nucleic acids by DTX3L, laying the groundwork for understanding its functional implications.
Negative regulation of APC/C activation by MAPK-mediated attenuation of Cdc20Slp1under stress
Authors: Sun Li , Chen Xuejin , Song Chunlin , Shi Wenjing , Liu Libo , Bai Shuang , Wang Xi , Chen Jiali , Jiang Chengyu , Wang Shuang-min , Luo Zhou-qing , Wang Ruiwen , Wang Yamei , Jin Quan-wen 
Published: April 2024

AbstractMitotic anaphase onset is a key cellular process tightly regulated by multiple kinases. The involvement of mitogen-activated protein kinases (MAPKs) in this process has been established inXenopusegg extracts. However, the detailed regulatory cascade remains elusive, and also it is unknown whether the MAPKs-dependent mitotic regulation is evolutionarily conserved in the single cell eukaryotic organism such as fission yeast (Schizosaccharomyces pombe). Here we show that two MAPKs inS. pombeindeed act in concert to restrain anaphase-promoting complex/cyclosome (APC/C) activity upon activation of the spindle assembly checkpoint (SAC). One MAPK, Pmk1, binds and phosphorylates Slp1Cdc20, the co-activator of APC/C. Phosphorylation of Slp1Cdc20by Pmk1, but not by Cdk1, promotes its subsequent ubiquitylation and degradation. Intriguingly, Pmk1-mediated phosphorylation event is also required to sustain SAC under environmental stress. Thus, our study establishes a new underlying molecular mechanism of negative regulation of APC/C by MAPK upon stress stimuli, and provides an unappreciated framework for regulation of anaphase entry in eukaryotic cells.One-sentence summaryInhibitory effect on the activation of anaphase promoting complex/cyclosome (APC/C) by MAPK Pmk1
HOIL1 mediates MDA5 activation through ubiquitination of LGP2
Authors: Cheng Deion , Zhu Junji , Liu GuanQun , Gack Michaela U. , MacDuff Donna A. 
Published: April 2024

AbstractThe RIG-I-like receptors (RLRs), RIG-I and MDA5, are innate sensors of RNA virus infections that are critical for mounting a robust antiviral immune response. We have shown previously that HOIL1, a component of the Linear Ubiquitin Chain Assembly Complex (LUBAC), is essential for interferon (IFN) induction in response to viruses sensed by MDA5, but not for viruses sensed by RIG-I. LUBAC contains two unusual E3 ubiquitin ligases, HOIL1 and HOIP. HOIP generates methionine-1-linked polyubiquitin chains, whereas HOIL1 has recently been shown to conjugate ubiquitin onto serine and threonine residues. Here, we examined the differential requirement for HOIL1 and HOIP E3 ligase activities in RLR-mediated IFN induction. We determined that HOIL1 E3 ligase activity was critical for MDA5-dependent IFN induction, while HOIP E3 ligase activity played only a modest role in promoting IFN induction. HOIL1 E3 ligase promoted MDA5 oligomerization, its translocation to mitochondrial-associated membranes, and the formation of MAVS aggregates. We identified that HOIL1 can interact with and facilitate the ubiquitination of LGP2, a positive regulator of MDA5 oligomerization. In summary, our work identifies LGP2 ubiquitination by HOIL1 in facilitating the activation of MDA5 and the induction of a robust IFN response.
Downregulation of Semaphorin 4A in keratinocytes reflects the features of non-lesional psoriasis
Authors: Kume Miki , Koguchi-Yoshioka Hanako , Nakai Shuichi , Matsumura Yutaka , Tanemura Atsushi , Yokoi Kazunori , Matsuda Shoichi , Nakamura Yuumi , Otani Naoya , Taminato Mifue , Tomita Koichi , Kubo Tateki , Wataya-Kaneda Mari , Kumanogoh Atsushi , Fujimoto Manabu , Watanabe Rei 
Published: April 2024

AbstractPsoriasis is a multifactorial disorder mediated by IL-17-producing T cells, involving immune cells and skin-constituting cells. Semaphorin 4A (Sema4A), an immune semaphorin, is known to take part in T helper type 1/17 differentiation and activation. However, Sema4A is also crucial for maintaining peripheral tissue homeostasis and its involvement in skin remains unknown. Here, we revealed that while Sema4A expression was pronounced in psoriatic blood lymphocytes and monocytes, it was downregulated in the keratinocytes of both psoriatic lesions and non-lesions compared to controls. Imiquimod application induced more severe dermatitis in Sema4A knockout (KO) mice compared to wild-type (WT) mice. The naïve skin of Sema4AKO mice showed increased T cell infiltration and IL-17A expression along with thicker epidermis and distinct cytokeratin expression compared to WT mice, which are hallmarks of psoriatic non-lesions. Analysis of bone marrow chimeric mice suggested that Sema4A expression in keratinocytes plays a regulatory role in imiquimod-induced dermatitis. The epidermis of psoriatic non-lesion and Sema4AKO mice demonstrated mTOR complex 1 upregulation, and the application of mTOR inhibitors reversed the skewed expression of cytokeratins in Sema4AKO mice. Conclusively, Sema4A- mediated signaling cascades can be triggers for psoriasis and targets in the treatment and prevention of psoriasis.
Transcript Isoform Diversity of Y Chromosome Ampliconic Genes of Great Apes Uncovered Using Long Reads and Telomere-to-Telomere Reference Genome Assemblies
Authors: Greshnova Aleksandra , Pál Karol , Martinez Juan Francisco Iturralde , Canzar Stefan , Makova Kateryna D. 
Published: April 2024

AbstractYchromosomes of great apes harborAmpliconicGenes (YAGs)—multi-copy gene families (BPY2,CDY,DAZ,HSFY,PRY,RBMY,TSPY,VCY, andXKRY) that encode proteins important for spermatogenesis. Previous work assembled YAG transcripts based on their targeted sequencing but not using reference genome assemblies, potentially resulting in an incomplete transcript repertoire. Here we used the recently produced gapless telomere-to-telomere (T2T) Y chromosome assemblies of great ape species (bonobo, chimpanzee, human, gorilla, Bornean orangutan, and Sumatran orangutan) and analyzed RNA data from whole-testis samples for the same species. We generated hybrid transcriptome assemblies by combining targeted long reads (Pacific Biosciences), untargeted long reads (Pacific Biosciences) and untargeted short reads (Illumina)and mapping them to the T2T reference genomes. Compared to the results from the reference-free approach, average transcript length was more than two times higher, and the total number of transcripts decreased three times, improving the quality of the assembled transcriptome. The reference-based transcriptome assemblies allowed us to differentiate transcripts originating from different Y chromosome gene copies and from their non-Y chromosome homologs. We identified two sources of transcriptome diversity—alternative splicing and gene duplication with subsequent diversification of gene copies. For each gene family, we detected transcribed pseudogenes along with protein-coding gene copies. We revealed previously unannotated gene copies of YAGs as compared to currently available NCBI annotations, as well as novel isoforms for annotated gene copies. This analysis paves the way for better understanding Y chromosome gene functions, which is important given their role in spermatogenesis.
High intracellular calcium amounts inhibit activation-induced proliferation of mouse T cells
Authors: Joseph Joel P , Kumar Tanisha , Chatterjee Kaushik , Nandi Dipankar 
Published: April 2024

AbstractOptimal T cell activation is critical to orchestrate adaptive immune responses. Calcium is critical for T cell activation and integrates signaling pathways necessary to activate key transcription factors. In fact, patients with calcium channelopathies are immunodeficient. Here, we investigated the effects of different concentrations of intracellular calcium on activation of mouse T cells. High intracellular calcium amounts inhibitedin vitroT cell proliferation as evidenced by a decreased cell cycling-to-hypodiploidy ratio in two models of activation: the combination of phorbol 12-myristate 13-acetate (PMA) and Ionomycin (an ionophore)/Thapsigargin (a SERCA inhibitor) or plate bound anti-CD3 and anti-CD28. High intracellular calcium amounts increased the production of reactive oxygen species (ROS) in T cells activated with PMA and Ionomycin and scavenging excess ROS using N-acetyl cysteine (NAC) or PEGylated superoxide dismutase (PEG-SOD) rescued the decrease in cycling-to-hypodiploidy ratio. To test the universality of our observations, we studied the effects of tert-Butylhydroquinone (tBHQ), a SERCA inhibitor and Nrf2 activator. tBHQ alone did not increase intracellular calcium amounts but increase was observed along with PMA. Also, tBHQ inhibited T cell activation in a dose-dependent manner in bothin vitromodels of T cell activation. Importantly, intraperitoneal injection of tBHQ ameliorated Dextran Sodium Sulfate (DSS)-induced colitis in mice as evidenced by rescue of colon length shortening and lower disease activity index. Overall, this study identifies high calcium amounts as a potential target to lower T cell activation. The implications of these observations are discussed as this strategy may be important in treatment of some autoimmune diseases.
A disease-associated PPP2R3C-MAP3K1 phospho-regulatory module controls centrosome function
Authors: Ganga Anil Kumar , Sweeney Lauren K. , Rubio Ramos Armando , Bishop Cassandra S. , Hamel Virginie , Guichard Paul , Breslow David K. 
Published: April 2024

SummaryCentrosomes have critical roles in microtubule organization and in cell signaling.1–8However, the mechanisms that regulate centrosome function are not fully defined, and thus how defects in centrosomal regulation contribute to disease is incompletely understood. From functional genomic analyses, we find here that PPP2R3C, a PP2A phosphatase subunit, is a distal centriole protein and functional partner of centriolar proteins CEP350 and FOP. We further show that a key function of PPP2R3C is to counteract the kinase activity of MAP3K1. In support of this model,MAP3K1knockout suppresses growth defects caused byPPP2R3Cinactivation, and MAP3K1 and PPP2R3C have opposing effects on basal and microtubule stress-induced JNK signaling. Illustrating the importance of balanced MAP3K1 and PPP2R3C activities, acute overexpression of MAP3K1 severely inhibits centrosome function and triggers rapid centriole disintegration. Additionally, inactivatingPPP2R3Cmutations and activatingMAP3K1mutations both cause congenital syndromes characterized by gonadal dysgenesis.9–15As a syndromicPPP2R3Cvariant is defective in centriolar localization and binding to centriolar protein FOP, we propose that imbalanced activity of this centrosomal kinase-phosphatase pair is the shared cause of these disorders. Thus, our findings reveal a new centrosomal phospho-regulatory module, shed light on disorders of gonadal development, and illustrate the power of systems genetics to identify previously unrecognized gene functions.
Structure and function of an intermediate GPCR-Gαβγ complex
Authors: Bi Maxine , Wang Xudong , Wang Jinan , Xu Jun , Sun Wenkai , Adediwura Victor Ayo , Miao Yinglong , Cheng Yifan , Ye Libin 
Published: April 2024

AbstractUnraveling the signaling roles of intermediate complexes is pivotal for G protein-coupled receptor (GPCR) drug development. Despite hundreds of GPCR-Gαβγ structures, these snapshots primarily capture the fully activated end-state complex. Consequently, a comprehensive understanding of the conformational transitions during GPCR activation and the roles of intermediate GPCR-G protein complexes in signaling remain elusive. Guided by a conformational landscape profiled by19F quantitative NMR (19F-qNMR) and Molecular Dynamics (MD) simulations, we resolved the structure of an unliganded GPCR-G protein intermediate complex by blocking its transition to the fully activated end-state complex. More importantly, we presented direct evidence that the intermediate GPCR-Gαsβγ complex initiates a rate-limited nucleotide exchange without progressing to the fully activated end-state complex, thereby bridging a significant gap in our understanding the complexity of GPCR signaling. Understanding the roles of individual conformational states and their complexes in signaling efficacy and bias will help us to design drugs that discriminately target a disease-related conformation.
ARID1A-BAF coordinates ZIC2 genomic occupancy for epithelial to mesenchymal transition in cranial neural crest lineage commitment
Authors: Barnada Samantha M. , de Gracia Aida Giner , Morenilla-Palao Cruz , López-Cascales María Teresa , Scopa Chiara , Waltrich Francis J. , Mikkers Harald M.M. , Cicardi Maria Elena , Karlin Jonathan , Trotti Davide , Peterson Kevin A. , Brugmann Samantha A. , Santen Gijs W. E. , McMahon Steven B. , Herrera Eloísa , Trizzino Marco 
Published: April 2024

AbstractThe BAF chromatin remodeler regulates lineage commitment including cranial neural crest cell (CNCC) specification. Variants in BAF subunits cause Coffin-Siris Syndrome (CSS), a congenital disorder characterized by coarse craniofacial features and intellectual disability. Approximately 50% of CSS patients carry variants in one of the mutually exclusive BAF subunits,ARID1A/ARID1B. WhileArid1adeletion in mouse neural crest causes severe craniofacial phenotypes, little is known about the role of ARID1A in CNCC specification. Using CSS patient-derivedARID1A+/-iPSCs to model CNCC specification, we discoveredARID1A-haploinsufficiency impairs epithelial to mesenchymal transition (EMT), a process necessary for CNCC delamination and migration from the neural tube. Furthermore, wild-type ARID1A-BAF regulates enhancers associated with EMT genes. ARID1A-BAF binding at these enhancers is impaired in heterozygotes while binding at promoters is unaffected. At the sequence level, these EMT enhancers contain binding motifs for ZIC2, and ZIC2 binding at these sites is ARID1A-dependent. When excluded from EMT enhancers, ZIC2 relocates to neuronal enhancers, triggering aberrant neuronal gene activation. In mice, deletion ofZic2impairs NCC delamination, whileZIC2overexpression in chick embryos at pre-migratory neural crest stages elicits ectopic delamination from the neural tube. These findings reveal a novel ARID1A-ZIC2 axis essential for EMT and CNCC delamination.