Analysis Of Markers Research Articles

Analysis and identification of ferroptosis-related diagnostic markers in rheumatoid arthritis.

Rheumatoid arthritis (RA) is an autoimmune, inflammatory joint disease. There is growing evidence that ferroptosis is involved in the pathogenesis of RA. This study aimed to search for diagnostic markers of ferroptosis in RA and to analyse the potential mechanisms and clinical value. RA-associated datasets were used from the publicly available GEO database. Three methods of machine learning were applied to screen biomarkers. The diagnostic efficacy of the results was also verified by receiver operating characteristic (ROC) curve, external dataset, qRT-PCR and Western blot. Enrichment analysis was performed in this process, while protein-protein interaction (PPI) analysis and immune infiltration correlation analysis were performed using biomarkers, and competing endogenous RNA (ceRNA) networks were constructed to search for prospective therapeutic targets. MMP13 and GABARAPL1 can be used as ferroptosis diagnostic genes in RA. The ROC curve and PPI result demonstrated that MMP13 and GABARAPL1 had an excellent diagnostic value. The results of signature genes in the external dataset, qRT-PCR and Western blot further confirm our findings. The enrichment analysis showed that p53, MAPK and NOD-like receptor signalling pathways may be involved in the process of ferroptosis in RA. In addition, two ferroptosis diagnostic genes in RA participate in the occurrence of ferroptosis in RA via oxidative stress, metabolism and immune response. Immune infiltration analysis showed that RA extensively infiltrated B cells, T cells, macrophages and other immune cells. Persistent immune activation may be an essential reason for the progression of ferroptosis in RA. We also obtained five potential therapeutic agents for RA and some long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) regulating ferroptosis diagnostic genes. Our study suggests that MMP13 and GABARAPL1, which are closely linked with oxidative stress and immunological modulation, can be used as ferroptosis-related potential diagnostic markers in RA and provide new clues regarding the diagnostic and therapeutic targets of ferroptosis in RA.

Predictive value analysis of albumin-related inflammatory markers for short-term outcomes in patients with In-hospital cardiac arrest

ABSTRACT Objective This study investigated the predictive value of albumin-related inflammatory markers for short-term outcomes in in-hospital cardiac arrest (IHCA) patients. Methods A linear mixed model investigated the dynamic changes of markers within 72 hours after return of spontaneous circulation (ROSC). Time-Dependent COX regression explored the predictive value. Mediation analysis quantified the association of markers with organ dysfunctions and adverse outcomes. Results Prognostic Nutritional Index (PNI) and RDW-Albumin Ratio (RAR) slightly changed (p > 0.05). Procalcitonin-Albumin Ratio (PAR1) initially increased and then slowly decreased. Neutrophil-Albumin Ratio (NAR) and Platelet-Albumin Ratio (PAR2) decreased slightly during 24-48 hours (all p＜0.05). PNI (HR = 1.646, 95%CI (1.033,2.623)), PAR1(HR = 1.69, 95%CI (1.057,2.701)), RAR (HR = 1.752,95%CI (1.103,2.783)) and NAR (HR = 1.724,95%CI (1.078,2.759)) were independently associated with in-hospital mortality. PNI (PM = 45.64%, 95%CI (17.05%,87.02%)), RAR (PM = 45.07%,95%CI (14.59%,93.70%)) and NAR (PM = 46.23%,95%CI (14.59%,93.70%)) indirectly influenced in-hospital mortality by increasing SOFA (central) scores. PNI (PM = 21.75%, 95%CI(0.67%,67.75%)) may also indirectly influenced outcome by increasing SOFA (renal) scores (all p＜0.05). Conclusions Within 72 hours after ROSC, albumin-related inflammatory markers (PNI, PAR1, RAR, and NAR) were identified as potential predictors of short-term prognosis in IHCA patients. They may mediate the adverse outcomes of patients by causing damages to the central nervous system and renal function.

Assessment of two benzylation strategies for the analysis of nerve-agent derived ethyl- and pinacolyl methyl phosphonic acids in sandy loam soil by GC–MS

Despite their prohibition by the Chemical Weapons Convention, nerve agents (NAs) remain in use against military and civilian targets. Due to their high reactivity, NAs readily degrade to phosphonic acids, making them important markers in the inspection of areas of presumed NA use. In this work, we assess the use of benzylation to modify ethyl- and pinacolyl methylphosphonic acids, degradation products of VX and Soman respectively, for their efficient detection in a soil matrix at ∼10 and ∼1 μg/g using GC–MS. The soil matrix, Sandy Loam (SL), was chosen for its ubiquitous nature, complex composition with silica particles embedded in clay, and low organic content. In this study, we demonstrate that benzylation via benzyl bromide yields a LOD = 25.6 ng/mL for benzylated-EMPA and LOD = 30.1 ng/mL for benzylated-PMPA. This is superior to the use of p-methoxybenzyl trichloroacetimidate in providing stable phosphonic acid ester derivatives for analysis. A base-modified procedure for p-methoxybenzylation was explored in this study yielding a LOD = 29.1 ng/mL for p-methoxybenzylated-EMPA and LOD = 39.8 ng/mL for p-methoxybenzylated-PMPA. Both benzylation pathways (benzyl bromide and p-methoxybenzyl trichloroacetimidate) can be used to yield phosphonic acid derivatives that provide further confirmation of these Soman and VX degradation products in soil samples in investigative scenarios. The work herein represents the first application of benzylation methods for the analysis of these NA markers in the acidic, silicon-based SL soil.

Genomic insights into the diversity of mango cultivars in Saudi Arabia: Fluorescent SSR marker analysis

ObjectivesThis study presents a comprehensive genetic analysis of twelve mango cultivars from Saudi Arabia, investigating their genetic diversity, population structure, and potential loci under selection. MethodsUtilizing ten SSR loci, we conducted a thorough assessment of allele variation, revealing diverse patterns across loci and defined genetic diversity among cultivars. ResultsNotably, certain loci exhibited extensive allele diversity, with Loci M02 and M06 displaying five and seven alleles, respectively, while others showed more limited profiles, such as Locus M01 with only two alleles. Surprisingly, clustering patterns based on genetic relatedness did not consistently align with anticipated geographic groupings, suggesting additional factors influencing genetic structure beyond geography. Further analysis of allele sharing provided insights into the genetic relatedness among cultivars, highlighting both close genetic affinities and significant divergence. Subsequent Allele Sharing Matrix analysis unveiled sub-clustering within clusters, revealing finer-scale genetic structures within the population. Assessment of genetic differentiation using Analysis of Molecular Variance demonstrated significant variations among clusters and individuals, with variations among clusters ranging from 3% to 53%. Additionally, testing for loci under selection identified potential markers, such as locus M11, with observed heterozygosity greater than expected (1.024) and a relatively low observed FST value of 0.2695. ConclusionsOverall, our findings contribute to the characterization of mango genetic resources in Saudi Arabia, providing crucial information for breeding programs, germplasm conservation efforts, and agricultural practices. By leveraging advanced genomic technologies and interdisciplinary collaborations, future research can delve deeper into the genetic basis of adaptive traits and agronomic characteristics, paving the way for innovative strategies to enhance mango production and sustainability in Saudi Arabia.

Testing the informativeness of Y-STR and mitochondrial DNA control region markers in an attempt to predict ancestry of World War II victims from Slovenian mass grave.

Identification of human remains is a challenge in forensic genetics without relatives or personal items available. In Slovenia, a Konfin II mass grave from the Second World War (WWII) was found, containing skeletal remains of 65 victims. The archival documents detailing victims' information describe 45 persons of which 33 could be considered Germanic and 12 Slavic. This study aims to check for concordance between the victim list and actual victims found by using uniparental markers to differentiate between Slavic and non-Slavic origin by attempting to infer ancestry by analyzing the control region (CR) of mitochondrial DNA (mtDNA) and Y-chromosomal STRs. Diaphyses of femurs were used as a DNA source. Next Generation Sequencing (NGS) technology was used for mtDNA- namely HID Ion Chef™ Instrument, Precision ID mtDNA Control Region Panel, and Ion GeneStudio™ S5 System. For the Y-chromosome, PowerPlex® Y23 System (Promega) kit and SeqStudio™ for human identification (HID) were used. European DNA Profiling mtDNA Population Database (EMPOP) and Y-Chromosome STR Haplotype Reference Database (YHRD) were searched for haplotype matches. Closest haplogroups were predicted using EMPOP, Y-DNA Haplogroup Predictor- NevGen, and Whit Athey's Haplogroup Predictor. Despite mitotypes being more diverse than Y-haplotypes, the Y-haplotypes had more database matches and more unequivocal differentiation between populations. 16 victims could be considered Slavic, 15 non-Slavic, and the remaining 34 had a rather scarce informativeness- either unclear or not providing any match. To address ancestry inference more comprehensively, analysis of autosomal ancestry informative markers as well as expansion on haploid markers will be conducted in future research.

The Politics of Identity Markers: A Critical Discourse Analysis of Pakistani English Newspapers

This study looked at how politicians come about with their political identities using identity markers to communicate their ideology(s) and ideas in political discourse. This study attempted to examine such identifying markers employed by Imran Khan (PTI), Fazal-ur-Rehman (JUI-F), Nawaz Sharif (PML-N), and Bilawal Bhutto (PPP) in Dawn and The Express Tribune news between 2018 and 2023. Data was obtained from the sites of a number of newspapers using a qualitative research method, using Fairclough's 3D model. Important markers of identification were phrases such as youth, corruption, accountability, and Naya Pakistan (new Pakistan), as well as cultural symbols, like flags and turbans. The ongoing use of pronouns, metaphors, and authoritative language by politicians helped to make societal topics such as power dynamics, corruption, and development lively. It is found that Fairclough's approach to the analysis of political identity markers can be put to good use.

Analysis of Tumor Proliferation Markers in Early-Stage Luminal Breast Cancer: A Comprehensive Study Using Mitotic Activity Index, Ki-67, and Phosphohistone H3 Expression.

Introduction and Aim: Routinely used proliferation markers such as mitotic activity index (MAI) and Ki-67 index show limited reproducibility due to high interobserver variability in breast cancer assessment. Phosphohistone H3 (PhH3), a novel proliferation marker, is gaining attention in breast cancer research. This study aimed to evaluate the inter-rater agreement among MAI, Ki-67, and PhH3 expressions in early-stage luminal breast cancer and assess the impact of replacing MAI with PhH3 index on tumor histological grading. Materials and Methods: Three pathologists assessed MAI, Ki-67, and PhH3 expressions in 66 early-stage luminal breast cancer specimens. Mitotic Activity Index was scored based on mitotic figures in an area of 2 mm2 while Ki-67 index utilized a 14% threshold for positively stained nuclei. Phosphohistone H3 expression cutoff was set at 13 positive cells per 2 mm2. The inter-rater agreement for the 3 variables was analyzed using Cohen kappa statistics. Results: Among the 3 parameters, the kappa score of the PhH3 expression reflected very strong agreement between the 3 observers (κ = 0.991, 0.907, and 0.916). Only moderate agreement was noted for MAI (κ = 0.898, 0.562, and 0.592) and substantial agreement for Ki-67 index (κ = 0.869, 0.673, and 0.678). Moreover, replacing MAI with PhH3 index led to upgrade of histological grade in 15% to 16% of patients. Conclusion: Our study demonstrated that PhH3 is a more reproducible proliferation marker than MAI and Ki-67. Incorporation of PhH3-based mitotic index in breast cancer grading might reduce the variation in the assessment of histological grade.

Abstract A004: Full spectrum isolation technique-based dual circulating rare cells analysis to maximize early cancer diagnosis

Abstract Liquid biopsy (LB) is essential in precision medicine, enabling real-time tumor monitoring through repeated sampling. However, its diagnostic sensitivity and accuracy are limited by the low detection rate of circulating tumor cells (CTCs). For example, in breast cancer, the CTC detection rate exhibited only 20%, significantly limiting its clinical utility. This limitation arises since existing technologies selectively isolate specific phenotypes of CTCs, failing to capture others, such as mesenchymal CTCs, which are associated with poor prognosis. These mesenchymal CTCs often evade detection due to their lack of EpCAM expression and high deformability, making difficult to isolate them with traditional filter or EpCAM based methods. Additionally, conventional LB methods fail to provide critical insights into the tumor microenvironment (TME). To overcome these limitations, we developed a novel approach that improves LB diagnostic sensitivity and accuracy by (1) creating a technology to isolate all types of CTCs, regardless of marker expression or size, and (2) simultaneously analyzing CTCs and circulating cancer-associated fibroblasts (cCAFs). This dual approach addresses the challenges of low sensitivity and accuracy by enabling the isolation of a broader range of CTCs, including mesenchymal types, and by incorporating cCAF analysis. cCAFs, derived from the TME and found in the bloodstream, provide valuable insights into the TME that are hard to obtain through traditional methods. Analyzing cCAFs alongside CTCs offers a more comprehensive understanding of tumor biology and its microenvironment, leading to improved diagnostic accuracy, especially in early-stage cancer. In our study involving 55 breast cancer patients, we demonstrated that while CTCs were undetectable in 42.9% of early-stage patients, cCAFs were detected in all cases and were found to be 10 times more abundant than CTCs. Importantly, for the first time, this study uncovers that the diverse CAF subtypes previously identified in tissue samples also exist in the bloodstream, highlighting their critical role in providing TME-related insights. In addition, cCAF subtypes vary with hormonal receptor expression, and their marker analysis allows for 80.9% accurate classification of breast cancer subtypes. We applied this approach to early-stage breast cancer patients, for whom diagnosis through liquid biopsy is particularly difficult, and found that the combined analysis of CTCs and cCAFs achieved a 18% improvement in accuracy, with the AUC increasing from 0.6817 to 0.8009 compared to CTC analysis alone. This innovative approach significantly enhances LB by simultaneously isolating all CTC subtypes and incorporating cCAF analysis, thereby offering a more sensitive and accurate diagnostic tool. Beyond improving breast cancer diagnostics, this method has the potential to be applied to other tumor types and could also prove valuable for prognosis prediction and therapeutic response monitoring, ultimately leading to better patient outcomes and more effective cancer treatment strategies. Citation Format: Hyeongjung Woo, Simon A Joosse, Frederike Harten, Felix Hilpert, Klaus Pantel, Minseok S. Kim. Full spectrum isolation technique-based dual circulating rare cells analysis to maximize early cancer diagnosis [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A004.

Evaluating Various Methods Used by Direct-to-Consumer Products Promising Personalized Nutrition

The increasing availability of direct-to-consumer personalized nutrition products has raised questions about their scope of personalization and added value compared to general dietary guidelines. This study evaluated eleven personalized nutrition products across six distinct methodologies, including DNA analysis, blood marker analysis, microbiome testing, blood glucose monitoring, questionnaire-based personalization, and consultation with a dietitian, comparatively evaluating their consistency and added value compared to general dietary guidelines. A 24-year-old obese male with no pre-existing illnesses was chosen as the case participant to represent a typical individual seeking personalized nutrition advice for weight loss. Significant variability and inconsistency were observed across and within the different methodologies, particularly in recommendations for energy intake and macronutrient distribution. Most advice was generic and aligned closely with existing dietary guidelines, with a notable lack of behavioral change strategies. This study underscores the challenges in personalized nutrition, highlighting the problem that current practices often fall short of delivering truly tailored recommendations. There is a critical need for clearer definitions, evidence-based guidelines, and more comprehensive approaches to fully realize the potential of personalized nutrition in improving individual health.

Cytogenetic and molecular identification of novel wheat-Elymus sibiricus addition lines with resistance to leaf rust and the presence of leaf pubescence trait

IntroductionEmerging new races of leaf rust (Puccinia triticina Eriks) are threatening global wheat (Triticum aestivum L.) production. Identifying additional resistance genes from all available gene pools is crucial to expanding wheat resistance to these virulent leaf rust races. Siberian wild rye (Elymus sibiricus L.) possesses numerous beneficial traits that can be valuable in wheat improvement. Three new wheat-E. sibiricus addition lines, O27-2 (BC8), O27-3 (BC12) and O193-3 (BC12), were developed through a backcrossing scheme in this study, using leaf rust field evaluations, molecular marker assays and cytogenetic analysis. MethodsThese three lines were derived from progeny of the bread wheat cultivar ‘Obriy’ (2n = 6x = 42, AABBDD) and partial octoploid amphiploid wheat-E. sibiricus (2n = 8x = 56, AABBDDStSt). Results and discussionThe lines (O27-2, O27-3 and O193-3) demonstrated strong specific leaf pubescence (hairiness) and resistance at the adult stage to a local population of leaf rust races. The response to leaf rust in these three lines significantly differed from that of the Lr24 gene, providing evidence for a distinct resistance mechanism associated with the 3St chromosome. This study is the first to report the transfer of an E. sibiricus chromosome into wheat that confers leaf rust resistance. Molecular marker analysis and genomic in situ hybridization confirmed that lines O27-2, O27-3 and O193-3 each possess one pair of E. sibiricus 3St chromosomes. The resistance gene was determined to be on the additional alien chromosome in these lines. Molecular markers (Xwmc221, Lr29F18, Sr24/Lr24) confirmed that the lines O27-2, O27-3, and O193-3 each contain a pair of E. sibiricus 3St chromosomes carrying leaf rust resistance genes. These findings demonstrate that the E. sibiricus 3St chromosome carries the leaf rust resistance gene and that the O27-2, O27-3, and O193-3 lines can serve as novel germplasm sources for introducing this resistance into wheat breeding programs. This study contributes to broadening the genetic diversity of resistance genes available for combating leaf rust in wheat.

Videos to study Interactions in AGEing (VIntAGE)

The Videos to study Interactions in AGEing (VIntAGE) corpus aims to investigate the complex relationship between language, cognition, and aging, focusing on verbal and non-verbal pragmatic markers in older persons with mild cognitive impairment (MCI). This multimodal and longitudinal corpus incorporates an analysis of gestural and verbal markers in discourse, aligned with neurolinguistic models. It provides a rich dataset for analyzing how aging impacts communicative competence in individuals with MCI. The VIntAGE corpus comprises approximately 18 hours of video recordings from 36 face-to-face interviews conducted by a close acquaintance of each of the nine women, all over 75 years old. Five participants were selected for in-depth analysis due to significant changes in their cognitive status. The participants underwent a series of semi-structured interviews over 15 months. The data were processed using transcription tools (for verbal discourse) and annotation tools (for gestures) and then subjected to Principal Component Analyses to manage each individual's diverse dataset and discursive modalities. The corpus includes the annotation of 6,351 verbal pragmatic markers (VPMs) and 8,044 non-verbal pragmatic markers (NVPMs). The data reveal an average decrease in MoCA scores from 23/30 to 20/30 over one year, highlighting cognitive decline's effects on verbal and non-verbal communication.

CNSC-06. NAVIGATING BRAIN CANCER: PATIENT STORIES AND MEDICAL BREAKTHROUGH

Abstract The complex interplay of genetic mutation, dys regulated signalling pathways, Epigenetic modifications and Interaction with tumor microenvironment are the major aggravating factors for Brain Tumors. Neurological complications involve Increased Intracranial pressure, Seizures, Motor and Sensory deficits, Aphasia, Hydrocephaly, Behavioral and personality changes, Cerebellar Symptoms. It is diagnosed in a sequential steps. Collecting good history from patient in primary then examining the patient all through and making some investigations to come to a final diagnosis. On asking history to a patient he complains of recurrent convulsions, Personality changes and some motor activity deficit and loss of few senses. On examination we can reveal defect in cranial nerve functions, Motor or sensory deficits, loss of reflexes and coordination. Then Investigations being the next step for diagnosis, We ask the patient to undergo for a test for Blood Biomarkers and CSF analysis in case of suspection of metastatic Brain tumors. Biopsy to obtain for Histopathological Examination. Histopathological examination is done for analysis of genetic mutations and molecular markers. The other imaging techniques like MRI, MRS, CT, PET, EEG are done. This gives a clear picture of what actually caused tumor and shows a way for treatment. Now the treatment involves both Medical and Surgical treatment. Medical treatment includes Chemotherapy along with radiation therapy where the drugs given are Temozolomide and targeted therapy included EGFR BRAF inhibitors for glioblastoma and low grade gliomas respectively. Tumor treating fields- Optune is given. Surgical treatment involves Craniotomy for removal of primary Brain tumors, Neuro Endoscopy if tumor is present in ventricles or base of Brain, Laser interstitial Thermal therapy for deap seated and recurrent brain tumors. This is all about the treatment and now the patient management is very important for further clinical help. Physical, occupational and speech therapy helps patients recover. Symptomatic approach for inoperable tumors Provide comprehensive support for patients nearing end of life, Emphasise confidence and Quality of life.

Impaired Bone Tissue Quality Associated with Inflammation in HIV-Immunological Non-Responders: A Cross-Sectional Analysis.

People with HIV (PWH) with poor immune response despite adequate antiretroviral treatment (ART) are susceptible to non-AIDS-related health issues. This study seeks to evaluate bone quality in Immunological Non-Responders (INRs) in comparison to those with proper immune response (IRs) using in vivo microindentation to quantify bone quality, in addition to conventional bone mineral density (BMD) evaluations. A cross-sectional study was conducted at Hospital del Mar in Barcelona from January 2019 to June 2023. Participants were matched in a 1:2-ratio (INRs:IR) based on age, sex, body mass index (BMI), and ART. Participants underwent bone quality assessment using in vivo microindentation, BMD and analysis of bone turnover and inflammation markers. Statistical analyses involved multivariable regression to adjust for potential confounding variables. A total of 159 PWH were included, 53 INRs and 106 IRs. INRs had worse bone quality, with lower median Bone Material Strength index (BMSi) compared to IRs (79 (76-87) vs. 86 (82-89); p<0.001), and similar BMD. INRs shown increased high-sensitive C-Reactive Protein levels with lower 25-(OH)-Vitamin D3. A significant negative correlation between inflammation and bone quality was found, especially noticeable in INRs. Multivariable linear regression shown that INR status is a major predictor of decreased bone quality, regardless of conventional risk factors. INRs condition is significantly associated with higher inflammatory levels, which may contribute to a deleterious effect on bone quality as measured by in vivo microindentation. Further studies are needed to confirm these results and to focus on non-AIDS comorbidities in this subgroup of PWH.

BIOM-09. CLINICAL EFFECT OF CD25 ON THE MTX-BASED CHEMOSENSITIVITY OF PRIMARY CENTRAL NERVUS SYSTEM LYMPHOMA

Abstract INTRODUCTION Primary central nervous system diffuse large B-cell lymphoma (PCNSL) is treated with chemotherapy, mainly with high-dose methotrexate (HD-MTX), but some cases are resistant to treatment and relapse in a short period. However, a biomarker that predicts the chemosensitivity and recurrence has not yet been identified. In this study, we used surface marker analysis of tumor tissue using flow cytometry to search for biomarkers related to chemosensitivity and early recurrence. METHODS From 2017 to 2023, 25 patients were diagnosed with PCNSL by biopsy in Kobe University Hospital, and flow cytometry of the biopsy tissues were performed. Cerebrospinal fluid (CSF) examination including sIL2R, β2MG, and IL-10 was performed in all cases before surgery, and treatment based on methotrexate was administered postoperatively. We retrospectively analyzed patient characteristics, flow cytometry data, treatment details, response rate, time to recurrence, and overall survival. RESULTS The mean age of the patients was 71 (range, 37-80) years. In univariate analysis, high expression level of CD25 (interleukin-2 receptor, IL-2R) in the tissue was associated with a lower response rate to MTX-based chemotherapy (Fisher’s exact test, p=0.0072) and had a tendency of shorter time to recurrence. However, there were no significant differences in progression-free survival and overall survival. There was no correlation between expression level of CD25 in tissue and sIL-2R in CSF. Expression level of sIL-2R in CSF was independent of CD25 expression in tissue. CONCLUSION There are some reports that CD25 can be a prognostic factor in follicular lymphoma, but no reports in PCNSL. Here, we suggested that high expression level of CD25 in PCNSL tissue was associated to chemoresistance and early recurrence. In cases of PCNSL with high CD25 expression level, additional whole-brain irradiation and transition to tirabrutinib be considered because there are possibilities of chemoresistance and early recurrence.

Therapeutic potential of mackerel-derived peptides and the synthetic tetrapeptide TVGF for sleep disorders in a light-induced anxiety zebrafish model

IntroductionAnxiety-like insomnia is a known risk factor for the onset and worsening of certain neurological diseases, including Alzheimer’s disease. Due to the adverse effects of current anti-insomnia medications, such as drug dependence and limited safety, researchers are actively exploring natural bioactive compounds to mitigate anxiety-like insomnia with fewer side effects. Mackerel (Pneumatophorus japonicus), a traditional Chinese medicine, is known for its tonic effects and is commonly used to treat neurasthenia. The use of mackerel protein extract has been shown to effectively improve symptoms of light-induced anxiety-like insomnia in a zebrafish model.MethodsThis study examines the effects of mackerel bone peptides (MW &amp;lt; 1 kDa, MBP1) and the synthetic peptide Thr-Val-Gly-Phe (TVGF) on light-induced anxiety-like insomnia in zebrafish. The evaluation is conducted through behavioral observation, biochemical marker analysis, and gene transcriptome profiling.ResultsMBP1 significantly alleviated abnormal hyperactivity and restored neurotransmitter levels (dopamine and γ-aminobutyric acid) to normal. Moreover, it mitigated oxidative stress by reducing reactive oxygen species production and malonaldehyde levels, while enhancing antioxidant enzyme activities (superoxide dismutase and catalase). This was further attributed to the regulation of lipid accumulation and protein homeostasis. Furthermore, MBP1 ameliorated sleep disturbances primarily by restoring normal expression levels of genes involved in circadian rhythm (per2 and sik1) and visual function (opn1mw2, zgc:73075, and arr3b). Molecular docking analysis indicated that TVGF exhibited good affinity for receptors linked to sleep disturbances, including IL6, HTR1A, and MAOA. TVGF exhibited sedative effects in behavioral assays, mainly mediated by regulating the normal expression of genes associated with circadian rhythm (cry1bb, cry1ba, per2, per1b and sik1), visual function (opn1mw1, gnb3b, arr3b, gnat2), purine metabolism (pnp5a), and stress recovery (fkbp5).DiscussionThese findings suggest that MBP1 and TVGF could be promising therapies for light-induced anxiety-like insomnia in humans, offering safer alternatives to current medications. Additionally, the regulation of genes related to circadian rhythm and visual perception may be a key mechanism by which MBP1 and TVGF effectively relieve anxiety-like insomnia.

Enhancing the Biological Properties of Organic–Inorganic Hybrid Calcium Silicate Cements: An In Vitro Study

(1) Background: This study aimed to enhance the biological properties of hydraulic calcium silicate cements (HCSCs) by incorporating organic and inorganic components, specifically elastin-like polypeptides (ELPs) and bioactive glass (BAG). We focused on the effects of these composites on the viability, migration, and osteogenic differentiation of human periodontal ligament fibroblasts (hPDLFs). (2) Methods: Proroot MTA was supplemented with 1–5 wt% 63S BAG and 10 wt% ELP. The experimental groups contained various combinations of HSCS with ELP and BAG. Cell viability was assessed using an MTT assay, cell migration was evaluated using wound healing and transwell assays, and osteogenic activity was determined through Alizarin Red S staining and a gene expression analysis of osteogenic markers (ALP, RUNX-2, OCN, and Col1A2). (3) Results: The combination of ELP and BAG significantly enhanced the viability of hPDLFs with an optimal BAG concentration of 1–4%. Cell migration assays demonstrated faster migration rates in groups with 2–4% BAG and ELP incorporation. Osteogenic activity was the highest with 2–3% BAG incorporation with ELP, as evidenced by intense Alizarin Red S staining and the upregulation of osteogenic differentiation markers. (4) Conclusions: The incorporation of ELP (organic) and BAG (inorganic) into HCSC significantly enhances the viability, migration, and osteogenic differentiation of hPDLFs. These findings suggest that composite HCSC might support healing in destructed bone lesions in endodontics.

Development of Microsatellite Markers from Transcriptome of Eriocheir sinensis and Their Application in Multiplex PCR Panels

Background: The development of genetic markers is crucial for aquaculture and population genetics. Polyallelic microsatellite markers are widely used for breeding and family management of Eriocheir sinensis (E. sinensis), contributing to the sustainable development of the industry. Methods: Through transcriptome analysis, 100 highly polymorphic SSR markers were identified and developed. From these, four multiplex PCR panels, each containing 16 markers, were constructed and applied to parentage assignment in six E. sinensis families. Results: The number of alleles (Na) ranged from 7 to 21, the observed heterozygosity (Ho) varied from 0.356 to 0.951, and the expected heterozygosity (He) varied from 0.656 to 0.909. The polymorphism information content (PIC) ranged from 0.633 to 0.900. The CERVUS analysis showed that using only the two most informative multiplex PCR panels, assignment success rates exceeded 90.34%, with 100% accuracy in assigning offspring to parents when three or four panels were used. Conclusions: This method significantly enhances the efficiency of microsatellite marker analysis, which provides a convenient and cost-effective method for breeding programs and family management of E. sinensis.

The Interaction of Apical Periodontitis, Cigarette Smoke, and Alcohol Consumption on Liver Antioxidant Status in Rats

This study aimed to investigate the impact of alcohol (A), secondhand cigarette smoking (ShS), and their combined effect on liver antioxidant activity and hepatic damage in rats with induced apical periodontitis (AP). Thirty-five female Wistar rats were randomly allocated into five groups (n = 7): (1) control (rats without ShS, alcoholic diet, or AP), (2) control-AP (induced AP only), (3) ShS-AP (ShS exposure and induced AP), (4) A-AP (alcoholic diet and induced AP), and (5) A+ShS-AP (alcoholic diet, ShS exposure, and induced AP). Alcohol was administered through semi-voluntary intake, while ShS exposure involved the daily inhalation of cigarette smoke. The experimental period lasted 8 weeks, with AP induction occurring in the 4th week following molar pulp exposure. Liver samples were collected post-euthanasia for histomorphometric and antioxidant marker analyses. All AP-induced groups exhibited increased liver sinusoidal dilation compared to the control group (p &lt; 0.05). AP significantly reduced total antioxidant capacity (FRAP) across all groups (p &lt; 0.05). In AP-induced groups, FRAP levels were further decreased in ShS-AP and A+ShS-AP compared to control-AP (p &lt; 0.05). AP also led to a decrease in the glutathione defense system (p &lt; 0.05). Rats with alcohol exposure (A-AP and A+ShS-AP) showed reduced glutathione peroxidase activity (p &lt; 0.05). Glutathione reductase activity was comparable in the control and control-AP groups (p &gt; 0.05), but significantly decreased in the alcohol and ShS-exposed groups (p &lt; 0.05). Apical periodontitis can relate to morphological changes in the liver’s sinusoidal spaces and impairment of liver’s antioxidant capacity of rats, particularly when combined with chronic alcohol consumption and exposure to cigarette smoke.

Development and characterization of a novel wheat-rye T2DS·2DL-2RL translocation line with high stripe rust resistance

Rye (Secale cereale L.), a close relative of common wheat, represents a valuable genetic resource for enhancing the disease resistance of common wheat. Introducing novel rye-derived genes into wheat can potentially improve disease resistance. In this study, we successfully developed a novel wheat-rye derivative line LCR4 through hybridization between hexaploid triticale line Currency and common wheat cultivar Jimai 22 (JM22). We confirmed that LCR4 was a T2DS·2DL-2RL translocation line via comprehensive molecular cytogenetic analyses, including genomic in situ hybridization, multi-color fluorescence in situ hybridization, molecular marker analysis, and wheat SNP-arrays genotyping. Notably, upon inoculation with Puccinia striiformis f. sp. tritici (Pst) race V26 at the seedling stage and mixed Pst races at the adult stage, LCR4 exhibited robust resistance against stripe rust infection at both stages. Subsequent genetic analysis further elucidated that the translocated 2RL chromosome segment is responsible for this resistance. Consequently, LCR4 harboring elite agronomic traits can be effectively employed in breeding programs against stripe rust.

Mir-509-3p targets SLC25A13 to regulate ferroptosis and protect retinal endothelial cells in diabetic retinopathy.

Diabetic retinopathy (DR) is a major complication of diabetes that leads to vision impairment. The aim of this study was to investigate the regulatory role of miR-509-3p in DR, focusing on its interaction with SLC25A13 and its impact on retinal endothelial cell function, oxidative stress, apoptosis, and ferroptosis. HRVECs were cultured in high-glucose (HG) conditions to establish an in vitro DR model. miR-509-3p mimics and inhibitors were transfected into HRVECs to assess their effects on SLC25A13 expression, cell viability, apoptosis, reactive oxygen species (ROS) levels, and ferroptosis markers. A luciferase reporter assay and RNA immunoprecipitation were used to confirm the binding of miR-509-3p to SLC25A13 mRNA. For in vivo validation, agomiR-509-3p was injected into the vitreous of DR mice, and retinal thickness, pathological damage, and apoptosis were evaluated. Ferroptosis-related markers (GPX4, TlR4, ASCL4) were analyzed in HRVECs to explore the mechanism of miR-509-3p in regulating ferroptosis. In vitro, miR-509-3p significantly decreased SLC25A13 expression, resulting in enhanced HRVEC viability, reduced apoptosis, and lower ROS levels under HG conditions. Overexpression of SLC25A13 reversed these protective effects, while miR-509-3p knockdown exacerbated oxidative stress and apoptosis. In vivo, agomiR-509-3p increased retinal thickness, reduced pathological damage, and decreased apoptosis in DR mice. Ferroptosis marker analysis revealed that miR-509-3p upregulated GPX4 expression and downregulated TlR4 and ASCL4, whereas SLC25A13 overexpression reversed these effects, further linking miR-509-3p to the regulation of ferroptosis. miR-509-3p exerts a protective effect in DR by targeting SLC25A13, reducing oxidative stress, apoptosis, and ferroptosis in retinal endothelial cells. These findings highlight the potential of miR-509-3p as a therapeutic target for DR management.