bluuebunny/crossref_metadata_2025 huggingface dataset:
Elucidating Sex-Specific Immune Profiles in a Breast Cancer Model
Authors: Hargrove-Wiley Ebony , Obodo Dora , Bindeman Wendy , Fingleton Barbara
Published: December 2024

Breast cancer is commonly thought of as a “women’s disease”. However, men are increasingly diagnosed with the disease, and their mortality rates are disparately higher than those of female patients. The abundance and composition of the immune microenvironment are determinants of breast cancer progression and survival. It is well documented that there are sex-specific differences in the immune response to several diseases, including various cancers. However, the effects of these differences in the context of breast cancer remain to be explored. This study demonstrates sex differences in the hormonal and immune landscape of the MMTV-PyMT transgenic murine model of female and male ER+ breast cancer using single-cell RNA sequencing (scRNA-Seq), whole-slide immunohistochemistry, and flow cytometry. Mammary tumors of transgenic male mice had increased estrogen receptor alpha expression and enriched nuclear binding signatures compared to female tumors. In the tumor immune compartment, male mice had lower intratumoral leukocyte infiltration. Yet, scRNA-Seq analysis reveals a more immunostimulatory microenvironment and increased antitumor immune populations in the primary and metastatic lungs as compared to transgenic females. Despite a more favorable innate immune profile, the metastatic burden was increased in male mice. Our data support a sex-dependent immune response in mammary carcinoma associated with the tumor, and likely host, hormonal environment. With emerging therapeutics targeting the tumor immune microenvironment, characterizing immune profiles is critical for optimizing their use in all breast cancer patients.
Hypoxia-Inducible Factor-2α Promotes Liver Fibrosis by Inducing Hepatocellular Death
Authors: Mooli Raja Gopal Reddy , Mukhi Dhanunjay , Watt Mikayla , Nagati Veerababu , Reed Sara M. , Gandhi Nikita K. , Oertel Michael , Ramakrishnan Sadeesh K.
Published: December 2024

The activation of hypoxia-inducible factors (HIF)-1α and 2α in the liver is closely linked to the progression of fatty liver diseases. Prior studies indicated that disrupting hepatocyte HIF-2α attenuates diet-induced hepatic steatosis, subsequently decreasing fibrosis. However, the direct role of hepatocyte HIF-2α in liver fibrosis has not been addressed. Hepatic HIF-2α expression was examined in mouse model of carbon tetrachloride (CCl4)-induced liver fibrosis. Conditional hepatocyte Hif-2α knockout mice were employed to investigate the role of hepatocyte HIF-2α in fibrosis. Markers of apoptosis, proliferation, inflammation, and fibrosis were assessed through biochemical, molecular, and histological analyses. We found an induction of HIF-2α in CCL4-injected liver injury and fibrosis mouse models. Hepatocyte-specific deletion of HIF-2α attenuated stellate cell activation and fibrosis, with no significant difference in inflammation. Disrupting hepatocyte HIF-2α led to reduced injury-mediated hepatocellular apoptosis. Surviving hepatocytes exhibited hypertrophy, which was strongly associated with the activation of c-JUN signaling. Our study demonstrates a direct role of hepatocyte HIF-2α in liver fibrosis by promoting hepatocyte apoptosis. The reduction in apoptosis and induction of hepatocyte hypertrophy following HIF-2α disruption is closely linked to enhanced c-JUN signaling, a survival mechanism in response to liver injury. These findings highlight HIF-2α as a potential therapeutic target for liver fibrosis.
Growth, Quality, and Nitrogen Metabolism of Medicago sativa Under Continuous Light from Red–Blue–Green LEDs Responded Better to High Nitrogen Concentrations than Under Red–Blue LEDs
Authors: Chen Ren , Chen Yanqi , Lin Kunming , Ding Yiming , Liu Wenke , Wang Shurong
Published: December 2024

Alfalfa is a widely grown forage with a high crude protein content. Clarifying the interactions between light quality and nitrogen level on yield and nitrogen metabolism can purposely improve alfalfa productivity in plant factories with artificial light (PFAL). In this study, the growth, quality, and nitrogen metabolism of alfalfa grown in PFAL were investigated using three nitrate-nitrogen concentrations (10, 15, and 20 mM, labeled as N10, N15, and N20) and continuous light (CL) with two light qualities (red–blue and red–blue–green light, labeled as RB-C and RBG-C). The results showed that the adaptation performance of alfalfa to nitrogen concentrations differed under red–blue and red–blue–green CL. Plant height, stem diameter, leaf area, yield, Chl a + b, Chl a, Chl b, crude protein contents, and NiR activity under the RB-CN15 treatment were significantly higher than RB-CN10 and RB-CN20 treatments. The RB-CN20 treatment showed morphological damage, such as plant dwarfing and leaf chlorosis, and physiological damage, including the accumulation of proline, H2O2, and MDA. However, the difference was that under red–blue–green CL, the leaf area, yield, and Chl a + b, carotenoid, nitrate, and glutamate contents under RBG-CN20 treatment were significantly higher than in the RBG-CN10 and RBG-CN15 treatments. Meanwhile, the contents of soluble sugar, starch, and cysteine were significantly lower. However, the crude protein content reached 21.15 mg·g−1. The fresh yield, dry yield, stomatal conductance, leaf area, plant height, stem diameter, crude protein, GS, and free amino acids of alfalfa were positively correlated with increased green light. In addition, with the increase in nitrogen concentration, photosynthetic capacity, NiR, and GOGAT activities increased, promoting growth and improving feeding value. The growth, yield, photosynthetic pigments, carbon, nitrogen substances, and enzyme activities of alfalfa were significantly affected by the interaction between nitrogen concentration and light quality, whereas leaf/stem ratio and DPPH had no effect. In conclusion, RB-CN15 and RBG-CN20 are suitable for the production of alfalfa in PFAL, and green light can increase the threshold for the nitrogen concentration adaptation of alfalfa.
Robust In Vitro Models for Studying Parkinson’s Disease? LUHMES Cells and SH-SH5Y Cells
Authors: Keighron Cameron Noah , Avazzedeh Sahar , Quinlan Leo R.
Published: December 2024

As our population ages, there is an increased unmet clinical need surrounding neurodegenerative diseases such as Parkinson’s disease (PD). To tackle this ever-increasing problem, we must ensure that the cell models that we use to develop therapeutics in vitro are robust, reliable, and replicable. In this study, we compared SH-SY5Y cells with LUHMES cells in response to 6-Hydroxydopamine (6OHDA) and 1-Methyl-4-phenylpyridinium (MPP+), two common Parkinson’s insults used in in vitro analysis. Both these cell types have apparent dopaminergic phenotypes, which could aid us in understanding their potential in this race to novel therapies. The LUHMES cells showed consistent dopaminergic (DA) expression through tyrosine hydroxylase (TH) positivity, alongside depleted ATP levels and elevated reactive oxygen species (ROS) production, whereas the SH-SH5Y cells displayed resilience to both chemical insults, raising questions about their utility in accurately modelling PD pathology. Further electrophysiological analysis revealed comparable firing rates and ion channel signalling between both cell types; however, LUHMES cells demonstrated stronger calcium signalling responses, further supporting their use as a more robust PD model. While SH-SY5Y cells showed some adaptability in vitro, their inconsistent DA phenotype and limited response to chemical insults limit their suitability for advanced research, suggesting that LUHMES cells should and must take their place as a hallmark in Parkinson’s disease research.
Non-Helicobacter pylori Helicobacters, a Treatable Provocateur of Parkinson’s Disease: Hypothesis, Evidence and Species Specificity
Authors: Wang Wenjing , Smith Melvyn , Ellis Richard , Savio Antonella , Nevel Amanda , Umamahesan Chianna , Pavlidis Polychronis , Hayee Bu’ Hussain , Taylor David , Young Allan H. , Charlett André , Dobbs Sylvia M. , Dobbs R. John
Published: December 2024

Epidemiological and eradication trial evidence indicates that Helicobacter pylori, a major causative factor in peptic ulcer and gastric cancer, is a driver of the hypokinesia of Parkinson’s disease (PD). Psychological (cognitive impairment, depression and anxiety) and gastrointestinal (peptic ulceration and constipation) PD features can precede the symptomatic onset of motor features by decades. We hypothesise that the non-H. pylori Helicobacters (NHPH), which have farm, companion and wild animals as their main hosts, can have a role in PD aetiopathogenesis. In those occupationally at risk of NHPH infection, we address whether there is increased mortality with PD, or depression or suicide. Our systematic review gave evidence that occupational exposure to animals/their products is associated with excess mortality with PD. Indeed, whilst livestock farming increased the risk, crop farming decreased it. Moreover, excess mortality from non-Hodgkin lymphoma in livestock farmers is compatible with NHPH being causal. Our scoping review showed that farmers, veterinarians and abattoir workers have an increased risk of depression and suicide; whether their depression is associated with being down the pathway to PD and/or the presence of Helicobacter infection needs investigation. Regarding Helicobacter species specificity, the link between the presence of NHPH in gastric biopsy and PD was described using a ureA polymerase chain reaction (PCR) assay, targeting the most-commonly named NHPH, H. suis. We describe its redesign and optimisation as a probe-based PCR, confirming the exclusion of H. pylori but not H. suis specificity (additionally identifying 6 species of a 22-NHPH-species panel). The exploration of the zoonotic hypothesis requires a non-invasive pan-Helicobacter PCR screen, allowing the detection and molecular grouping of Helicobacter species.
Homogalacturonans and Hemicelluloses in the External Glands of Utricularia dichotoma Traps
Authors: Płachno Bartosz J. , Kapusta Małgorzata , Feldo Marcin , Świątek Piotr
Published: December 2024

The Utricularia (bladderworts) species are carnivorous plants that prey mainly on invertebrates using traps (bladders) of leaf origin. On the outer surfaces of the trap, there are dome-shaped glands (capitate trichomes). Each such trichome consists of a basal cell, a pedestal cell, and a terminal cell. During the maturation of these external glands, there are changes in the cell wall of the terminal cell of the gland (deposited layers of secondary wall material). Thus, due to changes in the cell wall, these glands are excellent models for studying the specialization of cell walls. The main aim of this study was to check whether different cell wall layers in terminal gland cells have a different composition in the case of homogalacturonans (low-methylesterified HGs, fully de-esterified HGs, and galactan) and hemicelluloses (galactoxyloglucan, xyloglucan, and xylan). The antibodies were used against cell wall components (anti-pectins JIM5, JIM7, LM19, CCRC-M38, and LM5 and anti-hemicelluloses LM25, LM15, CCRC-M1, and CCRC-M138). The localization of the examined compounds was determined using immunohistochemistry techniques, Carbotrace 680, and Calcofluor White. Our study showed the presence of various components in the cell walls of external gland cells: methylesterified and demethylesterified homogalacturonans, galactan, xylan, galactoxyloglucan, and xyloglucan. In the terminal cell, the primary cell wall contains different pectins in contrast to the secondary wall material, which is rich in cellulose and hemicelluloses. We also found that the basal cell differs from the other gland cells by the presence of galactan in the cell wall, which resembles the epidermal cells and parenchyma of traps. A particularly noteworthy part of the cell wall functions as a Casparian strip in the pedestal cell. Here, we found no labeling with Carbotrace 680, possibly due to cell wall modification or cell wall chemical composition variation. We have shown that the apoplastic space formed by the cell walls of the terminal cell is mainly composed of cellulose and hemicelluloses (galactoxyloglucan and xyloglucan). This composition of the cell walls allows the easy uptake of components from the external environment. Our research supports the external glands’ function as hydropotens.
Pheromone-Binding Protein 1 Performs a Dual Function for Intra- and Intersexual Signaling in a Moth
Authors: Zhan Yidi , Zhang Jiahui , Xu Mengxian , Francis Frederic , Liu Yong
Published: December 2024

Moths use pheromones to ensure intraspecific communication. Nevertheless, few studies are focused on both intra- and intersexual communication based on pheromone recognition. Pheromone-binding proteins (PBPs) are generally believed pivotal for male moths in recognizing female pheromones. Our research revealed that PBP1 of Agriphila aeneociliella (AaenPBP1) serves a dual function in both intra- and intersexual pheromone recognition. Here, a total of 20 odorant-binding protein (OBP) family genes from A. aeneociliella were identified and subjected to transcriptional analysis. Among these, AaenPBP1 was primarily highly expressed in the antennae. Competitive fluorescence binding assays and molecular docking analyses demonstrated that AaenPBP1 exhibits a strong binding affinity for the female sex pheromone (Z)-9-Hexadecenyl acetate and the male pheromone 1-Nonanal. Notably, hydrogen bonds were observed between Ser56 and the ligands. The analysis of pheromone components and PBPs in lepidopteran lineage suggested that their strong and precise interactions, shaped by coevolution, may play a crucial role in facilitating reproductive isolation in moths. Our findings provide valuable insight into the functional significance of PBPs in invertebrates and support the development of behavioral regulation tools as part of an integrated pest management strategy targeting crambid pests.
Functional Study of PTSMAD4 in the Spermatogenesis of the Swimming Crab Portunus trituberculatus
Authors: Zhang Yu , Xiang Qiu-Meng , Mu Chang-Kao , Wang Chun-Lin , Hou Cong-Cong
Published: December 2024

Portunus trituberculatus holds significant economic value. The spermatogenesis is regulated by numerous signaling pathways. Among them, the TGF-β signaling pathway plays an important role in the development of testes and spermatogenesis. Smad4 is a Co-Smad protein that forms a complex with R-Smad to regulate the expression of target genes. The sperm structure in crustaceans differs greatly from that in mammals, with mature sperm lacking tails. Our previous studies have reported the function of R-Smad in the spermatogenesis of P. trituberculatus. In this study, we cloned the full-length cDNA sequence of PTSMAD4; immunofluorescence technology revealed that PTSMAD4 is expressed throughout all stages of spermatogenesis. We knocked down the expression of PTSMAD4 in P. trituberculatus using RNAi technology, and the immunofluorescence results show abnormal co-localization and weakened signals of PTSMAD4 and PTSMAD2. Additionally, transcriptome sequencing results enriched functional genes and pathways related to spermatogenesis. This study indicates that PTSMAD4 may participate in the spermatogenesis process through its involvement in signal transduction. This research not only lays the foundation for further study of the function of the TGF-β signaling pathway in spermatogenesis but also provides a theoretical basis for further investigation of the spermatogenesis mechanism in crustaceans.
Second Generation I-Body AD-214 Attenuates Unilateral Ureteral Obstruction (UUO)-Induced Kidney Fibrosis Through Inhibiting Leukocyte Infiltration and Macrophage Migration
Authors: Cao Qinghua , Foley Michael , Gill Anthony J. , Chou Angela , Chen Xin-Ming , Pollock Carol A.
Published: December 2024

Kidney fibrosis is the common pathological pathway in progressive chronic kidney disease (CKD), and current treatments are largely ineffective. The C-X-C chemokine receptor 4 (CXCR4) is crucial to fibrosis development. By using neural cell adhesion molecules as scaffolds with binding loops that mimic the shape of shark antibodies, fully humanized single-domain i-bodies have been developed. The first-generation i-body, AD-114, demonstrated antifibrotic effects in a mouse model of folic acid (FA)-induced renal fibrosis. The second-generation i-body, AD-214, is an Fc-fusion protein with an extended half-life, enhanced activity, and a mutated Fc domain to prevent immune activation. To investigate the renoprotective mechanisms of AD-214, RPTEC/TERT1 cells (a human proximal tubular cell line) were incubated with TGF-b1 with/without AD-214 and the supernatant was collected to measure collagen levels by Western blot. Mice with unilateral ureteral obstruction (UUO) received AD-214 intraperitoneally (i.p.) every two days for 14 days. Kidney fibrosis markers and kidney function were then analyzed. AD-214 suppressed TGF-b1-induced collagen overexpression in RPTEC/TERT1 cells. In UUO mice, AD-214 reduced extracellular matrix (ECM) deposition, restored kidney function, and limited leukocyte infiltration. In a scratch assay, AD-214 also inhibited macrophage migration. To conclude, i-body AD-214 attenuates UUO-induced kidney fibrosis by inhibiting leukocyte infiltration and macrophage migration.
Magnetic Isolation of the Linear Trinuclear Anion in [Cu(Him)6] {Cu(Him)4[Cu(μ-EDTA)(Him)]2}·6H2O (1) as the Novel Imidazolium(+) Salt (H2im)2[Cu(Him)4{(µ-EDTA)Cu(Him)}2]·2H2O (2)—A Comparative Look to Their Crystal Structures, Thermal, Spectral and Magnetic Properties and DFT Calculations
Authors: Belmont-Sánchez Jeannette Carolina , Choquesillo-Lazarte Duane , Frontera Antonio , Lezama Luis , Castiñeiras Alfonso , Niclós-Gutiérrez Juan
Published: December 2024

Inspired by the reported crystal structure of compound 1, we aimed to synthesize and determine the structure of compound 2, where two imidazolium (H2im+) ions serve as diamagnetic countercations. Here, we report the thermal stabilities, FT–IR, visible, and RSE spectra, as well as the magnetic properties of both compounds. In these structures, µ-EDTA acts as a pentadentate chelator for both terminal Cu centers within the centrosymmetric linear trinuclear anion. The Cu(µ-EDTA) chelates bind to the central Cu(Him)4 unit in subtly different ways: in compound 1, µ-EDTA has a free acetate arm and binds the central Cu(II) center through a syn,anti-carboxylate group. In contrast, in compound 2, the non-chelating acetate arm serves as a monodentate O-donor to the central Cu(II) atom, increasing the Cu(terminal)···Cu(central) distance from 6.08 Å in 1 to 6.80 Å in 2. Additionally, pairs of H2im+ ions in compound 2 display antiparallel π-stacking interactions. We conclude that the H2im+ counterions in compound 2 enable the magnetic isolation of the nearly identical trinuclear anion present in both compounds. DFT calculations further support the role of different interactions in stabilizing each crystal structure. In compound 2, dominant contributions from N–H···O hydrogen bonds and π-stacking interactions are accompanied by other, less conventional interactions, such as multiple C–H···O contacts and an O···CO(π-hole) interaction within the trinuclear anion.
Evaluation of Spliceosome Protein SmD2 as a Potential Target for Cancer Therapy
Authors: Li Jing , Li Peiyu , Brachtlova Tereza , van der Meulen-Muileman Ida H. , Dekker Henk , Kumar Vishal S. , Fransen Marieke , Bahce Idris , Felley-Bosco Emanuela , van Beusechem Victor W.
Published: December 2024

The core spliceosome Sm proteins are gaining attention as potential targets for cancer treatment. Here, we evaluate this, with focus on SmD2. A pan-cancer analysis including 26 solid tumor types revealed that the SmD2-encoding SNRPD2 gene was overexpressed in almost all cancers. In several cancers, high SNRPD2 expression was associated with a poor prognosis. To investigate the vulnerability of human cells to the loss of SmD2 expression, we silenced SNRPD2 using a short hairpin-expressing lentiviral vector in established cancer cell lines; in short-term cultured melanoma cells; and in several normal cell cultures, including cancer-associated fibroblasts cultured from non-small cell lung cancer resections. Additionally, we analyzed publicly available cell viability datasets for the dependency of cancer cell lines to SmD2 expression. Together, these studies clearly established SmD2 as a cancer-selective lethal target. Delving into genes with similar essentiality profiles to SNRPD2, we uncovered the intersected lethal stress between the loss of SmD2 and the loss of gene products participating in not only different mRNA processing steps including mRNA splicing, but also processes for coordinated protein production, as well as mitosis. Furthermore, we could correlate SNRPD2 expression to the responses of cancer cells to several FDA-approved anti-tumor drugs, especially to drugs inhibiting the cell cycle. Overall, our study confirms the anticipated role for targeting SmD2 in cancer treatment and reveals non-canonical SmD2 functions beyond mRNA splicing that could contribute to the dependency of cancer cells to high SNRPD2 expression.
Ezetimibe Enhances Lipid Droplet and Mitochondria Contact Formation, Improving Fatty Acid Transfer and Reducing Lipotoxicity in Alport Syndrome Podocytes
Authors: Kim Jin-Ju , Yang Eun-Jeong , Molina David Judith , Cho Sunjoo , Ficarella Maria , Pape Nils , Schiffer Josephin Elizabeth , Njeim Rachel , Kim Stephanie S. , Lo Re Claudia , Fontanella Antonio , Kaber Maria , Sloan Alexis , Merscher Sandra , Fornoni Alessia
Published: December 2024

Mitochondrial dysfunction is a critical factor in the pathogenesis of Alport syndrome (AS), contributing to podocyte injury and disease progression. Ezetimibe, a lipid-lowering drug, is known to inhibit cholesterol and fatty acid uptake and to reduce triglyceride content in the kidney cortex of mice with AS. However, its effects on lipid droplet (LD) utilization by mitochondria have not been explored. Transmission electron microscopy (TEM) and mitochondrial functional assays (ATP production, mitochondrial membrane potential, and citrate synthase activity) were used to investigate the impact of ezetimibe on LD–mitochondria contact formation and mitochondrial function in Col4a3KO (AS) and wildtype (WT) podocytes. TEM analysis revealed significant mitochondrial abnormalities in AS podocytes, including swollen mitochondria and reduced cristae density, while mitochondrial function assays showed decreased ATP production and lowered mitochondrial membrane potential. AS podocytes also demonstrated a higher content of LD but with reduced LD–mitochondria contact sites. Ezetimibe treatment significantly increased the number of LD–mitochondria contact sites, enhanced fatty acid transfer efficiency, and reduced intracellular lipid accumulation. These changes were associated with a marked reduction in the markers of lipotoxicity, such as apoptosis and oxidative stress. Mitochondrial function was significantly improved, evidenced by increased basal respiration, ATP production, maximal respiration capacity, and the restoration of mitochondrial membrane potential. Additionally, mitochondrial swelling was significantly reduced in ezetimibe-treated AS podocytes. Our findings reveal a novel role for ezetimibe in enhancing LD–mitochondria contact formation, leading to more efficient fatty acid transfer, reduced lipotoxicity, and improved mitochondrial function in AS podocytes. These results suggest that ezetimibe could be a promising therapeutic agent for treating mitochondrial dysfunction and lipid metabolism abnormalities in AS.
Exploring the Promoter Generation and Prediction of Halomonas spp. Based on GAN and Multi-Model Fusion Methods
Authors: Zhao Cuihuan , Guan Yuying , Yan Shuan , Li Jiahang
Published: December 2024

Promoters, as core elements in the regulation of gene expression, play a pivotal role in genetic engineering and synthetic biology. The accurate prediction and optimization of promoter strength are essential for advancing these fields. Here, we present the first promoter strength database tailored to Halomonas, an extremophilic microorganism, and propose a novel promoter design and prediction method based on generative adversarial networks (GANs) and multi-model fusion. The GAN model effectively learns the key features of Halomonas promoter sequences, such as the GC content and Moran’s coefficients, to generate biologically plausible promoter sequences. To enhance prediction accuracy, we developed a multi-model fusion framework integrating deep learning and machine learning approaches. Deep learning models, incorporating BiLSTM and CNN architectures, capture k-mer and PSSM features, whereas machine learning models utilize engineered string and non-string features to construct comprehensive feature matrices for the multidimensional analysis and prediction of promoter strength. Using the proposed framework, newly generated promoters via mutation were predicted, and their functional validity was experimentally confirmed. The integration of multiple models significantly reduced the experimental validation space through an intersection-based strategy, achieving a notable improvement in top quantile prediction accuracy, particularly within the top five quantiles. The robustness and applicability of this model were further validated on diverse datasets, including test sets and out-of-sample promoters. This study not only introduces an innovative approach for promoter design and prediction in Halomonas but also lays a foundation for advancing industrial biotechnology. Additionally, the proposed strategy of GAN-based generation coupled with multi-model prediction demonstrates versatility, offering a valuable reference for promoter design and strength prediction in other extremophiles. Our findings highlight the promising synergy between artificial intelligence and synthetic biology, underscoring their profound academic and practical implications.
Evasion of the Antiviral Innate Immunity by PRV
Authors: Wang Chenlong , Li Longxi , Zhai Xinyu , Chang Hongtao , Liu Huimin
Published: December 2024

Pseudorabies virus (PRV) establishes persistent latent infections by effectively evading the host’s antiviral innate immune response. PRV has developed sophisticated strategies to bypass immune surveillance through coevolution with its host. Currently, no effective vaccine exists to prevent or treat infections caused by emerging PRV variants, and the interactions between PRV and the host’s innate immune defenses remain incompletely understood. Nevertheless, ongoing research is uncovering insights that may lead to novel treatments and preventive approaches for herpesvirus-related diseases. This review summarizes recent advances in understanding how PRV disrupts key adaptors in immune signaling pathways to evade antiviral immunity. Additionally, we explored the intrinsic cellular defenses that play crucial roles in combating viral invasion. A deeper understanding of the immune evasion strategies of PRV could inform the development of new therapeutic targets and vaccines.
Comprehensive Gene Expression Analysis Using Human Induced Pluripotent Stem Cells Derived from Patients with Sleep Bruxism: A Preliminary In Vitro Study
Authors: Sato Taro , Yamaguchi Akihiro , Onishi Mayu , Abe Yuka , Shiga Takahiro , Ishikawa Kei-ichi , Baba Kazuyoshi , Akamatsu Wado
Published: December 2024

Sleep bruxism (SB) involves involuntary jaw movements during sleep and is potentially caused by motor neuronal hyperexcitability and GABAergic system dysfunction. However, the molecular basis remains unclear. In this study, we aimed to investigate changes in the expression of several genes associated with the pathophysiology of SB. Bulk RNA sequencing (bulk RNA-seq) and single-nucleus RNA sequencing (snRNA-seq) of neurons derived from patient and control human induced pluripotent stem cells (hiPSCs) were performed to comprehensively assess gene expression and cell type-specific alterations, respectively. Bulk RNA-seq revealed significant upregulation of calcium signaling-related genes in SB neurons, including those encoding G protein-coupled receptors and receptor-operated calcium channels. snRNA-seq confirmed the increased expression of GRIN2B (an N-methyl-D-aspartate receptor subunit) and CHRM3 (an M3 muscarinic acetylcholine receptor), particularly in glutamatergic and GABAergic neurons. These alterations were linked to hyperexcitability, with GRIN2B contributing to glutamatergic signaling and CHRM3 contributing to cholinergic signaling. These findings suggest that disrupted calcium signaling and overexpression of GRIN2B and CHRM3 drive neuronal hyperexcitability, providing insight into the pathophysiology of SB. Targeting these pathways may inform therapeutic strategies for SB treatment.
Metagenomic Analysis of the Buccal Microbiome by Nanopore Sequencing Reveals Structural Differences in the Microbiome of a Patient with Molar Incisor Hypomineralization (MIH) Compared to a Healthy Child—Case Study
Authors: Tynior Wojciech , Kłósek Małgorzata , Salatino Silvia , Cuber Piotr , Hudy Dorota , Nałęcz Dariusz , Chan Yuen-Ting , Gustave Carla , Strzelczyk Joanna Katarzyna
Published: December 2024

Molar incisor hypomineralization (MIH) is a qualitative developmental defect that affects the enamel tissue of permanent molars and can also occur in permanent incisors. Enamel affected by MIH has reduced hardness, increased porosity, and a higher organic content than unaffected enamel. These characteristics predispose the enamel to accumulation of bacteria and a higher prevalence of caries lesions. Through a groundbreaking metagenomic analysis of the buccal mucosal sample from a patient with MIH, we explored the intricacies of its microbiome compared to a healthy control using state-of-the-art nanopore long-read sequencing. Out of the 210 bacterial taxa identified in the MIH microbiome, we found Streptococcus and Haemophilus to be the most abundant genera. The bacteria with the highest read counts in the patient with MIH included Streptococcus mitis, Haemophilus parainfluenzae, Streptococcus pneumoniae, Rothia dentocariosa, and Gemella haemolysans. Our results revealed a striking contrast between healthy and MIH affected children, with a higher dominance and number of pathogenic species (S. pneumoniae, H. influenzae, and N. meningitidis) and reduced diversity in the MIH-affected patient. This distinct microbial profile not only sheds light on MIH-affected patients, but paves the way for future research, inspiring deeper understanding and larger scale studies.
Hesperetin Increases Lifespan and Antioxidant Ability Correlating with IIS, HSP, mtUPR, and JNK Pathways of Chronic Oxidative Stress in Caenorhabditis elegans
Authors: Wang Run-Jia , Ni Ya-Jing , Liu Yan-Qiang
Published: December 2024

Hesperetin (Hst) is a common citrus fruit flavonoid with antioxidant, anti-inflammatory, and anti-neurodegenerative effects. To explore the antioxidant and anti-aging effects and mechanisms of Hst, we induced chronic oxidative stress in Caenorhabditis elegans (C. elegans) using low-concentration H2O2 and examined its effects on lifespan, healthy life index, reactive oxygen species (ROS), antioxidant enzymes, and transcriptomic metrics. Hst significantly prolonged lifespan, increased body bending and pharyngeal pumping frequency, decreased ROS accumulation, and increased antioxidant enzyme activity in normal and stressed C. elegans. Hst significantly upregulated daf-18, daf-16, gst-2, gst-3, gst-4, gst-39, hsp-16.11, sip-1, clpp-1, and dve-1 and downregulated ist-1 and kgb-1 mRNAs in stressed C. elegans. These genes are involved in the insulin/insulin-like growth factor-1 signaling (IIS), heat shock protein (HSP), mitochondrial unfolded protein response (mtUPR), and c-Jun N-terminal kinase (JNK) pathways. In summary, Hst increases lifespan and antioxidant ability, correlating with these pathways, during chronic oxidative stress in C. elegans.
A Practical Format to Organize Cancer Constellations Using Innate Immune System Biomarkers: Implications for Early Diagnosis and Prognostication
Authors: Tobi Martin , Talwar Harvinder , Rossi Noreen F. , Lockette Warren , McVicker Benita
Published: December 2024

Cancer discovery is directed at the identification of a specific cancer type which allows for specific therapeutic interventions. Background/Objectives: Recently, similar immune checkpoint therapeutics have been applied with success across several cancer types, opening the field for other immune disruptive interventions that have practical applications. Methods: We have discovered an innate immune system (InImS) biomarker that allows for the characterization of allied cancer subtypes and outliers that might aid with diagnosis, treatment, and prognostication. Results: These InImS biomarkers are related to PD-L1 treatment outcomes and can be potentially manipulated by dietary means. Conclusions: The FERAD (ferritin–fecal p87) and absolute neutrophil/lymphocyte (aNLR) ratios are two such InImS biomarkers and we show herein, that they allow for the discovery of diagnosis and prognostication patterns, as demonstrated by this study.
Recent Advances in Marine-Derived Bioactives Towards Cancer Therapy
Authors: Tamzi Nafisa Nawar , Rahman Md Motiur , Das Subhadeep
Published: December 2024

The increase in cancer incidence in recent years necessitates urgent exploration of novel and alternative sources of natural bioactives for targeted cancer therapy. Approximately 75% of the Earth’s surface is covered by oceans, which are thought to harbor untapped physiologically active compounds with potential efficacy against cancer. Recently, a growing focus has been on isolating and investigating novel bioactive compounds derived from marine sources. Bioactive metabolites with diverse chemical structures, isolated from various marine species such as algae, mollusks, and actinomycetes, demonstrate potential efficacy against a wide range of cancers. To our knowledge, this is one of the articles that has reviewed recent papers on the application of marine-derived bioactives in targeted cancer therapy. This study aims to showcase some of the most current developments in targeted cancer therapy with various bioactives that have been identified from marine sources.
Guardians of the Genome: Iron–Sulfur Proteins in the Nucleus
Authors: Novoa-Aponte Lorena , Leon-Torres Andres , Philpott Caroline C.
Published: December 2024

Iron–sulfur (Fe-S) clusters are essential cofactors found in many proteins in the mitochondria, cytosol, and nucleus of the cell. These versatile cofactors may undergo reversible oxidation–reduction reactions to enable electron transfers; they may be structural and confer stability to a folded protein; they may be regulatory and transduce an iron signal that alters the function or stability of a recipient protein. Of the nearly 70 proteins described in mammalian cells that bind Fe-S clusters, about half localize exclusively or partially to the nucleus, where they are required for DNA replication and repair, telomere maintenance, transcription, mitosis, and cell cycle control. Most nuclear Fe-S cluster proteins interact with DNA, including DNA polymerases, primase, helicases, and glycosylases. However, the specific roles of the clusters in the enzymatic activities of these proteins and their interplay with DNA remain a matter of debate. Defects in the metallation of nuclear Fe-S proteins cause genome instability and alter the regulation of cell division and proliferation, which are hallmarks of various genetic diseases and cancers. Here, we provide an inventory of the nuclear Fe-S cluster-binding proteins and discuss cluster types, binding sites, the process of cluster acquisition, and the potential roles of the cluster in the function of the proteins. However, many questions remain unresolved. We highlight critical gaps in our understanding of cluster delivery to nuclear client proteins, the potential for cluster repair, and the mechanistic roles that clusters play in these enzymes. Taken together, this review brings the focus to the nucleus of the human cell as a hotspot for Fe-S cluster proteins and aims to inspire new research on the roles of iron in DNA metabolism and the maintenance of genome integrity.
Mitogenomics Provide New Phylogenetic Insights of the Family Apataniidae (Trichoptera: Integripalpia)
Authors: Ge Xinyu , Wang Jingyuan , Zang Haoming , Chai Lu , Liu Wenbin , Zhang Jiwei , Yan Chuncai , Wang Beixin
Published: December 2024

The family Apataniidae consists of two subfamilies, Apataniinae and Moropsychinae. Currently, there are 204 valid species of Apataniidae, which are widely distributed throughout the northern hemisphere. The larvae typically inhabit cold-water environments, and they serve as biological indicators for monitoring the health of freshwater ecosystems. The phylogenetic relationships within Apataniidae are not fully understood. Moreover, the available molecular data of Apataniidae are still limited. Herein, we provided the mitochondrial genomes of eight apataniid species and compared them with the published mitochondrial genomes of Apataniidae. The nine newly obtained sequences ranged from 15,070 bp to 16,737 bp in length. The results of the nonsynonymous with synonymous substitution rates displayed that ATP8 had the highest evolutionary rate, while COXI exhibited the lowest. The ND4L may be an effective molecular marker for the classification of the Apataniidae. Based on the published mitogenomes, we constructed a phylogenetic tree for Limnephiloidea and conducted a preliminary analysis of its advanced phylogeny. The ML and BI analyses recover the monophyly of Apataniidae and Limnephilidae. Except for PCG, BI tree based on other matrices consistently showed the topology: (Apataniana + (Moropsyche + (Apatidelia + Apatania))). The taxonomic status of Apatania and Apatidelia were also preliminarily explored. The mitochondrial genome of Apataniidae provides critical genomic resources for understanding the phylogenetic relationships of Apataniidae.
A Honey Bee In-and-Out Counting Method Based on Multiple Object Tracking Algorithm
Authors: Lei Chaokai , Lu Yuntao , Xing Zhiyuan , Zhang Jie , Li Shijuan , Wu Wei , Liu Shengping
Published: December 2024

The honey bee (Apis mellifera) is of great significance to both the ecological environment and human society, providing bee products and making a significant contribution to the pollination of crops [...]
Gla-Rich Protein Is Associated with Vascular Calcification, Inflammation, and Mineral Markers in Peritoneal Dialysis Patients
Authors: Marreiros Catarina , Viegas Carla , Guedes Anabela Malho , Silva Ana Paula , Águas Ana Catarina , Faísca Marília , Schurgers Leon , Simes Dina Costa
Published: December 2024

Background/Objectives: Vascular calcification (VC) is a crucial risk factor for cardiovascular diseases (CVD), particularly in chronic kidney disease (CKD) populations. However, the specific relationship between VC and end-stage renal disease (ESRD) patients undergoing peritoneal dialysis (PD) remains to be fully understood. The identification of new biomarkers to improve VC diagnosis and monitoring would significantly impact cardiovascular risk management in these high-risk patients. Gla-rich protein (GRP) is a VC inhibitor and an anti-inflammatory agent and thus is a potential VC marker in CKD. Here we explored the potential role of GRP as a marker for CVD and investigated the impact of VC in 101 PD patients. Methods: Circulating total Gla-rich protein (tGRP) was quantified in serum and in 24 h dialysate samples. VC score (VCS) was determined using the Adragão method. Results: Serum tGRP was negatively associated with VCS, serum calcium (Ca), phosphate (P), and high-sensitivity C-reactive protein (hsCRP), while it was positively associated with magnesium (Mg). A total of 35.6% of PD patients presented with extensive calcifications (VCS ≥ 3), and the lowest tGRP serum levels were present in this group (419.4 ± 198.5 pg/mL). tGRP in the 24 h dialysate was also negatively associated with VCS and with serum Ca and P. Moreover, serum Ca, P, and VCS were identified as independent determinants of serum tGRP levels. Conclusions: The association of serum tGRP with VC, mineral, and inflammation markers reinforces its potential use as a novel VC biomarker in CKD patients undergoing PD.
Percutaneous Atrial Septostomy in Adult Patients on Veno-Arterial Extracorporeal Membrane Oxygenation for Cardiogenic Shock: A Canadian Single-Center Experience
Authors: El Yamani Nidal , Mengi Siddhartha , Sénéchal Mario , Charbonneau Eric , Laflamme Maxime , Farjat-Pasos Julio , Rodés-Cabau Josep , Paradis Jean-Michel
Published: December 2024

Background/Objectives: Patients with cardiogenic shock on veno-arterial extracorporeal membrane oxygenation (VA-ECMO) frequently develop left ventricular (LV) distension and pulmonary edema due to an increased LV afterload. A balloon atrial septostomy (BAS) is a technique used to alleviate LV pressure and facilitate left atrial decompression. While primarily performed in pediatric populations, this procedure’s feasibility in adult patients is less studied. This study aimed to evaluate the procedural outcomes, including the safety and effectiveness, of BASs in adult patients with cardiogenic shock supported by VA-ECMO. Methods: This single-center retrospective study included 11 adult patients with cardiogenic shock on VA-ECMO, who underwent a BAS between 2012 and 2023. Multiple parameters were used to evaluate the global clinical impact of a BAS on patients with cardiogenic shock. Results: Between 2012 and 2023, 11 patients with cardiogenic shock on VA-ECMO underwent a BAS procedure in our institution. The mean time from the BAS to advanced therapy was 6.4 days. Procedural success was achieved in all patients with no complications. Nine patients (82%) had an improvement in PaO2/FiO2 24 h post-BAS procedure. All patients had an improvement in the pulmonary edema on the chest X-ray 24 to 48 h after the procedure, with clear radiography achieved in nine patients (82%) in a mean time of 7 days (range: 1.5–13 days). A total of five patients (45%) had in-hospital mortality due to non-procedural complications and the mortality timing from BAS was between 5 to 23 days. Among those discharged, all six patients were alive at the 1-year follow-up. Conclusions: A BAS is a feasible and safe technique for decompressing the left atrium in adult patients on VA-ECMO. It significantly improved pulmonary edema and oxygenation in most cases. Further studies with larger populations are needed to evaluate its impact on long-term outcomes.
Development and Initial Outcomes of the Interdisciplinary ‘Early Identification and Intervention for Infants Network’ (Ei3) in Los Angeles
Authors: Butera Christiana D. , Yeh Amy , Biniwale Manoj , Bloch Edward , Craddock Debi , Doyle Mary , Iyer Sai N. , Kretch Kari S. , Liu Nora , Mirzaian Christine B. , Sargent Barbara , Solano Priscilla , Swaine Margaret , Dusing Stacey C.
Published: December 2024

Background/Objectives: The Early Identification and Intervention for Infants (Ei3) Network is an interdisciplinary team dedicated to improving early detection and intervention of cerebral palsy (CP) in California. This paper describes the key (1) awareness-building and (2) capacity-building strategies utilized by the Ei3 Network in the first two years. Methods: Awareness-building methods included interactive conference discussions, resource deliverable creation, and the creation of a framework for dissemination. Capacity-building methods were hosting assessment training, gathering stakeholder feedback, and implementation training. All deliverables were created with a minimum of 3 review and revision cycles. We planned, hosted, and provided scholarships for training, including the Hammersmith Infant Neurological Examination (HINE), Prechtl’s GMA, and an implementation conference. Preliminary descriptive statistics and paired samples t-tests were performed on HINE training surveys. Results: Seven resource deliverables were created and distributed. A professional website, @steps2home.org, was launched. Online channels gained followers (146, Instagram; 198, X; 298, Mailchimp). Providers participated in various trainings: the Sykes Symposium on early detection and intervention (70), HINE (211), GMA Basic or Advanced (46), and Implementation Conference (six facilities from April 2023 to June 2024). HINE training participants reported increased ability to identify CP (p = 0.001), knowledge of (p = 0.004) early detection, ability to implement early detection guidelines (p < 0.001), and confidence in performing the HINE on the post-test (79.87%). Conclusions: The first two years of Ei3 resulted in increased awareness of the early detection guidelines through resources to help providers and families navigate complex care systems and dissemination through online and in-person strategies. We built capacity by training an increased number of providers to identify infants at risk of CP and implement the early detection guidelines. Engagement of stakeholders in focus groups, reviewing documents, and including a parent panel throughout the process increased the value of this work and should support the expansion of the network in the next year.
Association Between Serum Concentrations of (Certain) Metals and Type 2 Diabetes Mellitus
Authors: Tyczyńska Magdalena , Hunek Gabriela , Kawecka Weronika , Brachet Adam , Gędek Marta , Kulczycka Kinga , Czarnek Katarzyna , Flieger Jolanta , Baj Jacek
Published: December 2024

The findings regarding trace element concentrations in patients diagnosed with type 2 diabetes and healthy controls are inconsistent, and therefore, we determined to gather them in the form of a review to further indicate the need for more advanced knowledge development. In our study, we reviewed articles and studies that involved the topics of micronutrient and metal associations with the occurrence and development of type 2 diabetes. We mainly included works regarding human-based studies, but with limited research results, animal-based research was also taken into account. With some newer studies, we reached for initial assumptions of previous statements. The results indicated that higher serum levels of lead, cadmium, arsenic, bromine, barium, strontium, nickel, aluminum, calcium, copper, and ferritin are positively associated with diabetic prevalence. Both too-low and too-high levels of zinc, selenium, and magnesium may be connected to the development of diabetes. Chromium has the capability of insulin response modulation, with enhanced insulin-cell binding, and thus, lower serum levels of chromium can be found in diabetic patients. There are contradictory discoveries regarding manganese. Its supplementation can possibly cease the development of insulin resistance and type 2 diabetes. On the contrary, other studies reported that there is no such connection. Our work indicates that, as micronutrients play a significant role in the pathogenesis of metabolic disorders, more research regarding their bodily homeostasis and type 2 diabetes should be conducted.