Search Results (809)

Detailed analysis of gene expression by real time-quantitative polymerase chain reaction (RT-qPCR) has become a widespread method. To normalize the expression of target genes, this approach relies on constitutively expressed internal controls known as housekeeping genes (HKGs). Their proper selection is a critically important methodological step, since all the studied gene expression will be recalculated based on HKG expression. This concise review aims to discuss the selection of HKGs for endometrial cancer (EC) studies. We draw attention to the fact that the commonly used gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is unsuitable as a HKG for research on the normal endometrium, EC, as well as many other tissues. In contrast, accumulating evidence suggests that GAPDH is a pan-cancer marker and an EC marker. Work on GAPDH overexpression in EC in relation to overall and relapse-free survival is lacking. Both original research and overviews indicate that at least two HKGs should be used for target gene expression recalculations, a rarely applied technical aspect of final data processing. The insufficiently careful selection in many studies of only one HKG, e.g., GAPDH, can be held responsible for broad discrepancies in published results obtained by this RT-qPCR technique. We provide an account of the discrepancies reported for sex hormone receptors expression in EC. Achieving consensus on the selection and validation of HKGs for research on this cancer is of crucial importance. Ideally, this trusted gene combination should be universal for any EC histotype and grade, irrespective of the final anatomopathological result. Full article

Background: The COVID-19 pandemic provided a unique opportunity to evaluate how the human immune system responded to a novel pathogen and to determine whether immune responses initiated through natural infection differ from those elicited by vaccination against the same antigen. Here, we provide a comprehensive analysis of SARS-CoV-2 Spike (S-antigen)-reactive memory B cells (B_mem) elicited in previously immunologically naïve subjects following their first infection with the original Wuhan-Hu-1 (WH1)-like strain or their initial COVID-19 mRNA prime-boost regimen encoding the same WH1-S-antigen. In particular, we tested the hypothesis that the primary encounter of SARS-CoV-2 S-antigen in lung mucosal tissues during infection vs. intramuscular COVID-19 mRNA injection would elicit different B_mem responses. Methods: Cryopreserved peripheral blood mononuclear cell (PBMC) samples collected following primary infection with the WH1 strain or completion of the initial prime-boost vaccination regimen were tested in ImmunoSpot^® assays to assess the frequency, Ig class/subclass usage, and cross-reactivity of the S-antigen-reactive B_mem compartment; pre-pandemic blood draws served as naïve controls. Results: The B_mem repertoires generated post-infection vs. post-vaccination were found to be quite similar but with some subtle differences. In both cases, the prevalent induction of IgG1-expressing B_mem in similar frequencies was seen, ~30% of which targeted the receptor binding domain (RBD) of the WH1-S-antigen. Also, the extent of cross-reactivity with the future Omicron (BA.1) RBD was found to be similar for both cohorts. However, IgA⁺ B_mem were preferentially induced after infection, while IgG4⁺ B_mem were detected only after vaccination. Conclusions: B_mem elicited in naïve human subjects following SARS-CoV-2 infection or after WH1-S encoding mRNA vaccination were only subtly different, although the relevance of these differences as it relates to immune protection warrants further investigation. Our findings serve to illustrate the usefulness and feasibility of performing comprehensive monitoring of antigen-specific B cell memory in larger cohorts using the ImmunoSpot^® technique. Full article

Changes in the habitat of wild mallard ducks (Anas platyrhynchos L.) and thus in their diet can result in significant differences in the content of sodium and potassium in their tissues and organs. There is little data in the available literature regarding the qualitative analysis of mallard meat and organs. The aim of the study was to determine the sodium and potassium content in biological material (breast muscle, leg muscles, and liver) from wild mallards and the effect of sex and place of origin (hunting district) on these parameters. Sodium and potassium in the biological material were analyzed by inductively coupled plasma atomic emission spectroscopy (ICP-OES). The sodium and potassium levels in the tissues were shown to be influenced by the sex of the mallards and the site where they were harvested. Sodium content was significantly higher in the liver of male mallards than in females. In most cases, the tissues and organs of mallards harvested in the Siedlce hunting district had higher levels of sodium and potassium, apart from potassium content in the breast muscle. This may indicate greater abundance of these elements in this district. Full article

Background/Objectives: HER2, a critical diagnostic marker and therapeutic target in breast cancer, is a membrane receptor that forms diverse signaling complexes, the constituents of which have not been fully characterized in actual breast cancer tissues. Methods: In this study, we applied the Rapid Immunoprecipitation Mass Spectrometry of Endogenous Proteins (RIME) method, originally developed to explore transcription factor complexes, to identify the complexes formed by HER2 in HER2-positive breast cancer specimens. Results: Through our approach, we successfully identified multiple complex components, including MARCKS, a novel HER2-interacting partner, which we verified using both proximal ligation assay in cultured cells and immunohistochemistry in tissue sections. TCGA analysis further revealed that high MARCKS expression significantly correlates with ER negativity, as confirmed by multivariate analysis, suggesting its potential role as a prognostic indicator in aggressive breast cancer subtypes. Conclusions: These results demonstrate the capability of RIME to elucidate interactomes of membrane proteins such as HER2 in clinical tissue specimens. Furthermore, this study highlights its broader applicability beyond nuclear proteins, underscoring its potential for discovering novel prognostic and diagnostic clinical markers in diverse cancer types. Full article

Background/Objectives: This systematic review aims to synthesize recent studies on the integration of artificial intelligence (AI) into robotic surgery for oncological patients. It focuses on studies using real patient data and AI tools in robotic oncologic surgery. Methods: This systematic review followed PRISMA guidelines to ensure a robust methodology. A comprehensive search was conducted in June 2025 across Embase, Medline, Web of Science, medRxiv, Google Scholar, and IEEE databases, using MeSH terms, relevant keywords, and Boolean logic. Eligible studies were original research articles published in English between 2024 and 2025, focusing on AI applications in robotic cancer surgery using real patient data. Studies were excluded if they were non-peer-reviewed, used synthetic/preclinical data, addressed non-oncologic indications, or explored non-robotic AI applications. This approach ensured the selection of studies with practical clinical relevance. Results: The search identified 989 articles, with 17 duplicates removed. After screening, 921 were excluded, and 37 others were eliminated for reasons such as misalignment with inclusion criteria or lack of full text. Ultimately, 14 articles were included, with 8 using a retrospective design and 6 based on prospective data. These included articles that varied significantly in terms of the number of participants, ranging from several dozen to several thousand. These studies explored the application of AI across various stages of robotic oncologic surgery, including preoperative planning, intraoperative support, and postoperative predictions. The quality of 11 included studies was very good and good. Conclusions: AI significantly supports robotic oncologic surgery at various stages. In preoperative planning, it helps estimate the risk of conversion from minimally invasive to open colectomy in colon cancer. During surgery, AI enables precise tumor and vascular structure localization, enhancing resection accuracy, preserving healthy tissue, and reducing warm ischemia time. Postoperatively, AI’s flexibility in predicting functional and oncological outcomes through context-specific models demonstrates its value in improving patient care. Due to the relatively small number of cases analyzed, further analysis of the issues presented in this review is necessary. Full article

Objectives: Connective tissue diseases (CTDs) include a diverse group of systemic autoimmune conditions, among which interstitial lung disease (ILD) is acknowledged as a major determinant of prognosis. High-resolution computed tomography (HRCT) is the gold standard for ILD assessment. The distribution of HRCT patterns across CTDs remain incompletely defined. The objective of this systematic review is to synthesize available evidence regarding the prevalence of specific radiological patterns within CTD-ILDs and to assess whether specific patterns occur at different frequencies among individual CTDs. Methods: The inclusion criteria encompassed original human studies published in English between 2015 and 2024, involving adult participants (≥18 years) with CTD-ILDs assessed primarily by HRCT and designed as retrospective, prospective, or cross-sectional trials with extractable data. We systematically searched PubMed, Scopus, and Web of Science (January 2025). Risk of bias was evaluated using the Newcastle–Ottawa Scale (NOS) for cohort and case–control studies, and the JBI Critical Appraisal Checklist for cross-sectional studies. Data were extracted and categorized by HRCT pattern for each CTD, and then summarized descriptively and statistically. Results: We analyzed 23 studies published between 2015 and 2024, which included 2020 patients with CTD-ILDs. The analysis revealed non-specific interstitial pneumonia (NSIP) as the most prevalent pattern overall (36.5%), followed by definite usual interstitial pneumonia (UIP) (24.8%), organizing pneumonia (OP) (9.8%) and lymphoid interstitial pneumonia (LIP) (1.25%). HRCT distribution varied by CTD: NSIP predominated in systemic sclerosis, idiopathic inflammatory myopathies, and mixed connective tissue disease; UIP was most frequent in rheumatoid arthritis; LIP was more common in Sjögren’s syndrome. While global differences were statistically significant, pairwise comparisons often lacked significance, likely due to sample size constraints. Discussion: Limitations include varying risk of bias across study designs, heterogeneity in HRCT reporting, small sample sizes, and inconsistent follow-up, which may reduce precision and generalizability. In addition to the quantitative synthesis, this review offers a detailed description of each radiologic pattern mentioned above, illustrated by representative examples to support the recognition in clinical settings. Furthermore, it includes a brief overview of the major CTDs associated with ILD, summarizing their epidemiological data, risk factors for ILD and clinical presentation and diagnostic recommendations. Conclusions: NSIP emerged as the most common HRCT pattern across CTD-ILDs, with UIP predominating in RA. Although inter-disease differences were observed, statistical significance was limited, likely reflecting sample size constraints. These findings emphasize the diagnostic and prognostic relevance of HRCT pattern recognition and highlight the need for larger, standardized studies. Full article

The interventricular septum is recognized as the first region to undergo remodeling, and a septal bulge is described as an early echocardiographic sign of hypertensive heart disease. Using third-generation microscopic ultrasonography in an animal model, we validated, for the first time, that remodeling originates in the basal septum, presenting as basal septal hypertrophy (BSH), an early imaging biomarker, and subsequently progresses to other regions, leading to tissue dysfunction and heart failure. We have termed this finding the “stress septal sign” (Triple S) because a variety of stress stimuli, such as treadmill exercise or pressure overload from aortic banding in animals, induced BSH, a region with more intensive sympathetic innervation than the mid-apex. This finding also represents a conjunctive point between functional etiologies, such as hypertension, and emotional etiologies that precipitate acute stress cardiomyopathy. Microscopic analysis of the remodeling revealed that hemodynamic stress has a specific effect on cardiac geometry. The Triple S is associated with exercise-induced hypertension and high stress scores in patients with hypertension. Furthermore, three-dimensional segmental remodeling is more effective than cross-sectional measurements for detecting the impact of superimposed multiple stressors. A high-rate pressure product and blood pressure variability in patients exhibiting the Triple S should be managed comprehensively through an integrated approach to stress and hypertension to avoid high mortality in clinical practice. A precise etiologic evaluation of incidentally detected BSH may contribute to the early diagnosis of hypertensive disease. The integrated and timely management of stress and hypertension is important for patients presenting with the Triple S and high stress scores. This management strategy may provide a practical solution for avoiding the adverse hypertensive consequences of global remodeling and maladaptation to superimposed multiple stressors. Full article

Inflammatory bowel diseases (IBDs), such as ulcerative colitis, and Crohn’s disease are chronic idiopathic inflammatory diseases of the gastrointestinal system involving interaction between genetic and environmental factors mediating the occurrence of oxidative stress and inflammation. There is no permanent cure for IBD except long-term treatment or surgery (resection of the intestine), and the available agents in the long term appear unsatisfactory and elicit numerous adverse effects. To keep the disease in remission, prevent relapses and minimize adverse effects of currently used medicines, novel dietary compounds of natural origin convincingly appear to be one of the important therapeutic strategies for the pharmacological targeting of oxidative stress and inflammation. Therefore, it is imperative to investigate plant-derived dietary agents to overcome the debilitating conditions of IBD. In the present study, the effect of α-Bisabolol (BSB), a dietary bioactive monoterpene commonly found in many edible plants as well as important components of traditional medicines, was investigated in acetic acid (AA)-induced colitis model in rats. BSB was orally administered to Wistar male rats at a dose of 50 mg/kg/day either for 3 days before or 30 min after induction of IBD for 7 days through intrarectal administration of AA. The changes in body weight, macroscopic and microscopic analysis of the colon and calprotectin levels in the colon of rats from different experimental groups were observed on day 0, 2, 4, and 7. The levels of myeloperoxidase (MPO), a marker of neutrophil activation, reduced glutathione (GSH) and malondialdehyde (MDA), a marker of lipid peroxidation, and the levels of pro-inflammatory cytokines were measured. AA caused a significant reduction in body weight and induced macroscopic and microscopic ulcers, along with a significant decline of endogenous antioxidants (superoxide dismutase (SOD), catalase, and GSH), with a concomitant increase in MDA level and MPO activity. BSB significantly improved the AA-induced reduction in body weight, colonic mucosal histology, inhibited MDA formation, and restored antioxidant levels along with a reduction in MPO activity. AA also induced the release of pro-inflammatory cytokines such as interleukin-1 (IL-1), interleukin-23 (IL-23) and tumor necrosis factor-α (TNF-α). Furthermore, AA also increased levels of calprotectin, a protein released by neutrophils under inflammatory conditions of the gastrointestinal tract. BSB treatment significantly reduced the release of calprotectin and pro-inflammatory cytokines. The findings of the present study demonstrate that BSB has the potential to improve disease activity and rescue colonic tissues from damage by inhibiting oxidative stress, lipid peroxidation and inflammation. The findings are suggestive of the benefits of BSB in IBD treatment and substantiate its usefulness in colitis management, along with its gastroprotective effects in gastric ulcer. Full article

Osteosarcoma (OS) and extraskeletal osteosarcoma (EOS) in dogs exhibit histological similarities but differ in anatomical origin, which poses a challenge to diagnostic accuracy. We adopted a marker-first strategy to enhance molecular classification by selecting RUNX2, KPNA2, and SATB2, three validated immunohistochemical (IHC) markers, as primary targets. Bioinformatic screening identified the miR-30 family as the only miRNA group predicted to coordinately regulate RUNX2, KPNA2, and SATB2, justifying its prioritization for expression analysis. RT-qPCR on FFPE tissues from 14 OS, 19 EOS, and 10 healthy controls revealed that miR-30a was significantly downregulated in OS and inversely correlated with RUNX2 nuclear expression, confirmed by IHC. MiR-30e also showed high diagnostic accuracy, while miR-30b and miR-30c distinguished EOS from OS. Non-seed interaction modeling (i.e., outside the canonical “seed” region, spanning nucleotides 2–8 of the miRNA) suggested divergent regulatory affinities within the PI3K/AKT/RUNX2 axis among miR-30 family members. MiR-30a and miR-30e exhibited the highest diagnostic power (LR⁺ 7.7 and 6.8, respectively), supporting their role as biomarkers. These results highlight a miR–30–centered regulatory axis with relevance for diagnosis and molecular stratification of canine osteogenic tumors. Full article

Background and Clinical Significance: The 45,X0/46,XY mosaic karyotype is categorized as a disorder of sex development and can lead to atypical sexual development. Latent mosaicism involving Y chromosomal segments may be much more prevalent than previously assumed, according to a growing number of findings. This primarily depends on how sensitive cytogenetic methods are—such as traditional karyotype screening, FISH methods, or molecular analyses. Case Presentation: We present the case of a 10-week-old infant with hermaphroditic external genitalia. During pregnancy, ultrasonography revealed severe fetal development difficulties, including severe widespread edema. An abnormal 45,X0/46,XY mosaic karyotype was discovered during a genetic amniocentesis conducted during the 16th week of pregnancy. The infant was born in average general condition at 39 + 6 weeks of gestation. Physical examination of the infant revealed features of facial dysmorphia, webbed neck, and hermaphroditic external genitalia. The testicle was palpable on the left side, but the gonad was absent on the right. Laboratory tests revealed a typical hormonal profile of the mini-puberty period in boys. Moreover, a hormone panel and thyroid ultrasound were performed; congenital hypothyroidism was diagnosed. Three separate independent sources of biological material were used in cytogenetic analysis to determine the karyotype: skin fibroblasts (to confirm tissue mosaicism), oral epithelial cells (FISH), and peripheral blood lymphocytes. It showed that a mosaic occurred very early in embryogenesis by confirming the existence of karyotypes 45,X and 46,XY in various tissues (mosaic tissue distribution). Conclusions: Tissue mosaicism should be compared to the analysis of tissues from other embryonic origins, including blood and oral tissue. Support for gender identity and treatment decisions, including the prediction of the future risk of gonadoblastoma, as well as multidisciplinary care, is necessary. Full article

Squamosa Promoter-Binding Protein Like (SPL) is a critical transcription factor that plays a significant role in regulating plant growth and development. Mining the coconut SPL family offers valuable insights into the regulation of important agronomic traits, including the length of the juvenile phase. In this study, 25 CnSPLs were identified and were classified into eight subfamilies. Analysis of gene structure and conserved protein motifs indicated a high conservation of CnSPLs within the same subfamilies; however, variations in protein structure and gene length were observed across different subfamilies. Gene expansion analysis indicated that most gene members within subfamilies originated from duplications of the same genomic segment, and transposable element insertion contributed to the divergence of gene sequences within these subfamilies. Characterization of the miR156 target sequence in SPL transcripts revealed that subfamilies IV to VIII contained these sequences, while subfamilies I to III did not. In both coconut and 14 other plant species, some SPLs lost their miR156-binding loci due to gene structure variations. The gene expression profiles revealed significant divergence between miR156-targeted and non-targeted CnSPLs; the former exhibited low expression levels in the endosperm, while the latter showed comparable expression across all tissues. Notably, CnSPL15A demonstrated steadily increasing expression levels in leaves throughout successive leaf primordia and significantly promoted flowering when overexpressed in Arabidopsis. Transient expression assays and 5′ RACE confirmed that CnSPLs are targeted by miR156. This study establishes a foundation for investigating the evolutionary characteristics of CnSPLs and provides a theoretical framework for analyzing the functions of key CnSPLs involved in the coconut flowering control pathway. Full article

Reactive oxygen species (ROS) are fundamental components found in cells that exist in an oxygen environment. While they are often viewed as detrimental metabolic byproducts that can harm cells, leading to aging and cell death, they can also play a role in cellular regulatory processes and have beneficial effects. One of the main ROS present in all cells is hydrogen peroxide (H₂O₂), which can function as a signaling molecule in extra- and intracellular signaling. To enhance our understanding of how various enzymes regulate cellular H₂O₂ levels, we created a mathematical model of H₂O₂ neutralization and performed computer simulations to estimate the neutralization efficiency in various types of cells. Data on gene expression for genes participating in this process were incorporated into the calculations, along with the regulation of enzymes in oxidation and reduction processes. The conducted simulations demonstrate that cells originating from different tissues utilize systems neutralizing H₂O₂ variously, which results in differences in H₂O₂ cellular levels. The simulation findings suggest that the differences in radiosensitivity seen in various cancer cell types may be linked to their effectiveness in scavenging H₂O₂. Analysis of results from model simulations for colorectal, lung, and breast cancer cell lines indicated that radiosensitive cell lines exhibited elevated levels of H₂O₂, attributed to the reduced efficiency of neutralizing enzymes. By highlighting cell-type-specific differences in H₂O₂ neutralization, our findings may contribute to a deeper understanding of redox regulation in cancer cells and reveal new potential correlations with radioresistance. Full article

Background/Objectives: Our work was designed to study the physicochemical properties, safety profile, pharmacokinetics, and prophylactic efficacy of an original iodine–dextrin-based pharmaceutical formulation (PA), both alone and in combination with azithromycin (AZ), in a murine model of LPS-induced sepsis. Methods/Results: UV–vis and ¹H-NMR spectroscopy confirmed the formation of a stable iodine–dextrin complex, with triiodide anions stabilized by hydrogen bonding and donor–acceptor interactions. No clinical signs of acute toxicity were observed at doses up to 5000 mg/kg, and subacute administration (62.5 and 125 mg/kg) showed no adverse effects on hematological or biochemical parameters. A mild, non-pathological enlargement of thyrocytes and parallel increases in TSH, T3, and T4 levels were observed at 125 mg/kg, consistent with physiological adaptation to iodine. Pharmacokinetic analysis revealed high oral bioavailability (~92%), prolonged half-life (~21 h), and wide tissue distribution with low clearance. In the sepsis model, pretreatment with AZ+PA alleviated clinical symptoms, maintained body weight, and significantly improved hematological parameters, reducing WBCs and CRP levels. The combination also decreased plasma IL-6 and TNF-α concentrations more effectively than either agent alone, indicating a synergistic anti-inflammatory effect. Histological analysis confirmed that PA, particularly in combination with AZ, mitigated LPS-induced tissue injury in the liver, kidney, and lungs. Conclusions: These findings suggest that PA is a safe, bioavailable compound with immunomodulatory properties that enhance azithromycin’s protective effects during systemic inflammation. This supports its potential use as a prophylactic agent in clinical settings, such as preoperative immune modulation to prevent sepsis-related complications. Full article

Background: Mesenchymal stem cells (MSCs) are a promising tool in regenerative medicine due to their ability to secrete paracrine factors that modulate tissue repair. Extracellular vesicles (EVs) released by MSCs contain bioactive molecules (e.g., mRNAs, miRNAs, proteins) and play a key role in intercellular communication. Methods: This study compared the transcriptomic profiles (mRNA and miRNA) of equine MSCs derived from adipose tissue (AT-MSCs), bone marrow (BM-MSCs), and ovarian fibroblasts (as a differentiated control). Additionally, miRNAs present in EVs secreted by these cells were characterized using next-generation sequencing. Results: All cell types met ISCT criteria for MSCs, including CD90 expression, lack of MHC II, trilineage differentiation, and adherence. EVs were isolated using ultracentrifugation and validated with nanoparticle tracking analysis and flow cytometry (CD63, CD81). Differential expression analysis revealed distinct mRNA and miRNA profiles across cell types and their secreted EVs, correlating with tissue origin. BM-MSCs showed unique regulation of genes linked to early development and osteogenesis. EVs contained diverse RNA species, including miRNA, mRNA, lncRNA, rRNA, and others. In total, 227 and 256 mature miRNAs were detected in BM-MSCs and AT-MSCs, respectively, including two novel miRNAs per MSC type. Fibroblasts expressed 209 mature miRNAs, including one novel miRNA also found in MSCs. Compared to fibroblasts, 60 and 92 differentially expressed miRNAs were identified in AT-MSCs and BM-MSCs, respectively. Conclusions: The results indicate that MSC tissue origin influences both transcriptomic profiles and EV miRNA content, which may help to interpret their therapeutic potential. Identifying key mRNAs and miRNAs could aid in future optimizing of MSC-based therapies in horses. Full article

Autism spectrum disorder (ASD) is a complex neurodevelopmental condition with early-life origins. Maternal obesity has been associated with increased ASD risk, yet the mechanisms and timing of susceptibility remain unclear. Using a mouse model combining in vitro fertilization (IVF) and embryo transfer, we separated the effects of pre-conception and gestational obesity. We found that maternal high fat diet (HFD) exposure prior to conception alone was sufficient to induce ASD-like behaviors in male offspring—including altered vocalizations, reduced sociability, and increased repetitive grooming—without anxiety-related changes. These phenotypes were absent in female offspring and those exposed only during gestation. Cortical transcriptome analysis revealed dysregulation and isoform shifts in genes implicated in ASD, including Homer1 and Zswim6. Whole-genome bisulfite sequencing of hippocampal tissue showed hypomethylation of an alternative Homer1 promoter, correlating with increased expression of the short isoform Homer1a, which is known to disrupt synaptic scaffolding. This pattern was specific to mice with ASD-like behaviors. Our findings show that pre-conceptional maternal obesity can lead to lasting, isoform-specific transcriptomic and epigenetic changes in the offspring’s brain. These results underscore the importance of maternal health before pregnancy as a critical and modifiable factor in ASD risk. Full article