Background:

Current treatments for traumatic brain injury (TBI) commonly focus on symptom management and lack strategies to prevent or delay the development of neurodegenerative processes. The lack of understanding of TBI mechanisms hinders the development of effective and safe approaches for anti-inflammatory therapy and the stimulation of regeneration in damaged and/or lost neurons. The study of the blood system in TBI may provide new biomarkers for the development of neurodegenerative processes and methods for regulating neuroinflammation and neuroregeneration.

Materials and Methods:

Male ICR mice, 12-14 weeks old, were used in this experiment. Traumatic brain injury was induced by focal impact using a weight-drop model [Chakraborty, N., Hammamieh, R., Gautam, A., et al., 2021]. The neurological status of the mice was assessed before and at 6 hours, as well as on days 1, 3, 7, 14, 21, and 42 post-injury. Histological examination (hematoxylin and eosin staining), immunohistochemical staining, and analysis of the expression of astrocyte, mature, and immature neuron markers in the brain were performed. The content of hematopoietic cells in the blood and bone marrow was studied staining by the May–Grünwald–Giemsa staining method, along with the quantitative and qualitative composition of hematopoietic niches [Crocker P., Gordon S., 1985]. Additionally, an in vitro analysis of hematopoietic and progenitor cells was performed. The sympatholytic drug reserpine was used to modulate neuroinflammation and neurogenesis.

Results:

Reserpine, via decreasing hematopoietic activity, reduced brain damage, prevented neuroinflammation, and facilitated neurogenesis in mice subjected to TBI.

Conclusion:

Pharmacological blockade of the sympathetic component of hematopoiesis regulation may serve as a basis for developing new approaches to reduce neuroinflammation and promote neurogenesis in TBIs.

G-protein-coupled receptors (GPCRs) are integral membrane proteins that mediate critical physiological processes by translating extracellular signals into intracellular responses. Among them, the β2-Adrenergic Receptor (β2-AR) plays a significant role in smooth-muscle relaxation, bronchodilation, and cardiovascular regulation, making it a primary target for treating conditions such as asthma, hypertension, and chronic obstructive pulmonary disease (COPD) [1]. While synthetic drugs are widely used to target β2-AR, they often lead to adverse effects and drug resistance, necessitating the exploration of alternative therapeutic approaches [2].

This study utilized molecular docking methods with AutoDock 4.6 to investigate the interactions of natural compounds, including ephedrine, quercetin, catechin, and resveratrol, with β2-AR. The receptor's 3D structure was retrieved from the Protein Data Bank (PDB ID: 2RH1) [3], and docking studies evaluated the binding energies, hydrogen bonding, and other stabilizing interactions between the ligands and the receptor's active site. Ephedrine was observed to form hydrogen bonds with key residues, consistent with previous studies [4]. Quercetin exhibited strong binding interactions with the β2-AR's active site, confirming its potential as a natural inhibitor [5].

These findings highlight the promise of natural compounds in modulating GPCR activity, particularly β2-AR, as safe and effective alternatives to synthetic drugs. The application of computational methods like molecular docking further emphasizes their utility in identifying bio-inspired strategies for drug discovery targeting GPCR-related diseases.

References

[1] Johnson M. Paediatr Respir Rev. 2006;7(Suppl 1):S3–S7.

[2] Liu Y, He Y, Wang K. Phytother Res. 2019;33(3):571–582.

[3] Cherezov V, Rosenbaum DM, Hanson MA, et al. Science. 2007;318(5854):1258–1265.

[4] Kim YJ, et al. J Mol Graph Model. 2017;76:1-9.

[5] Wang L, et al. Biomed Pharmacother. 2021;134:111066.

Background: G-protein-coupled receptors (GPCRs) are a diverse group of membrane proteins that mediate critical physiological processes by converting extracellular signals into intracellular responses. The β2-Adrenergic Receptor (β2-AR), a key GPCR, plays a pivotal role in smooth-muscle relaxation, bronchodilation, and cardiovascular function, making it a therapeutic target for conditions such hypertension or asthma. Diketopiperazines (DPKs), as the simplest cyclic peptides, have emerged as promising scaffolds for inhibiting protein interactions and modulating receptor activity, offering a novel, appealing therapeutic approach with potentially fewer side effects compared to small-molecule inhibitors.

Methods: In this study, five DPK derivatives were obtained from PubChem and evaluated for their binding affinity to 3D structure of β2-AR (PDB ID = 2RH1) through molecular docking studies using Autodock 4.6 and MGLTools. Each compound’s binding energy and hydrogen bond formation were assessed to determine their interaction efficiency.

Results: Among the five compounds, tryptophan–proline diketopiperazine (compound 3) exhibited the highest binding affinity with a binding energy of -5.89 kcal/mol and formed two hydrogen bonds. The enhanced interaction is attributed to the aromatic nature of tryptophan, which promotes strong π-π stacking interactions, and the rigidity of proline, which allows for optimal fitting within the receptor's binding pocket. The hydrophobic interactions further stabilized the complex.

Conclusion: This study highlights that diketopiperazine (DPK) derivatives, particularly tryptophan–proline diketopiperazine, are promising inhibitors of the β2-Adrenergic Receptor (β2-AR), a key G-protein-coupled receptor (GPCR). The compound’s aromaticity and rigidity enhance receptor interaction, providing insights into the design of peptide-based inhibitors for β2-AR and other GPCR-related diseases, with potential for improved specificity and fewer side effects.

Introduction. Postoperative infectious complications, especially in critically ill patients, have a high mortality risk. The early diagnosis of such complications and mortality prediction in the intensive care unit (ICU) are urgent tasks.

Objective. We aimed to identify and measure tyrosine and phenylalanine metabolites that are clinically significant for the diagnosis of infectious complications and to carry out mortality prediction in critically ill patients.

Materials and methods. A total of 298 patients were examined, including patients with acute abdominal surgical diseases (n = 58); patients after cardiac surgery (n = 79) or neurosurgery (n = 82); and other high-risk patients on the day of admission to the ICU, regardless of the main pathology (n = 79). The level of aromatic metabolites in the blood serum or cerebrospinal fluid samples of patients was determined by means of gas chromatography.

Results. In patients with acute surgical diseases of the abdominal cavity with bacterial–inflammatory complications (n=35), the median serum levels of phenyllactic (PhLA) and 4-hydroxyphenyllactic acid (p-HPhLA) were 2.5 (p<0.001) and 1.5 (p=0.048) times higher, respectively, than those of patients without an infectious process (n=23). Six hours after cardiac surgery, in patients who subsequently developed infectious complications (n=26), the median serum levels of p-HPhLA and the sum of sepsis-associated metabolites were 1.4 (p=0.010) and 1.6 (p=0.002) times higher, respectively, than those of patients who did not develop complications (n=36). In post-neurosurgical patients with signs of secondary meningitis (n=30), the median level of p-HPhLA in the cerebrospinal fluid was 2.7 times higher (p<0.001) than that of patients without signs of secondary meningitis (n=52). On admission to the ICU, non-survived patients (n=35) had at least 5-fold higher median serum levels of p-HPhLA and other sepsis-associated metabolites (p<0.001) compared with survivors (n=44).

Conclusions: The levels of sepsis-associated metabolites of tyrosine and phenylalanine can be used to predict the development of infectious complications and mortality in high-risk patients.

Leishmaniasis, caused by around 20 species of Leishmania, affects millions of people in endemic areas, particularly in tropical and subtropical regions. However, current treatments face challenges due to significant side effects and increasing drug resistance. This study aims to develop new anti-leishmanial therapies by focusing on the threonine biosynthesis pathway, due to its uniqueness in parasites. This pathway allows the parasite to synthesize threonine, an essential amino acid, while humans must obtain it through their diet. Therefore, targeting this pathway presents a promising opportunity for selective drug action. Homoserine kinase (HSK), a rate-limiting enzyme in the threonine biosynthesis pathway, was identified as a promising therapeutic target due to its crucial role in converting L-homoserine to ortho-phospho homoserine, a precursor to threonine. Using a multifaceted approach—including in silico virtual screening, in vitro biochemical assays, and parasite culture techniques—we screened the Maybridge chemical library and discovered two HSK inhibitors, RH00038 and S02587, with strong potential for further development. Both inhibitors demonstrated significant efficacy, inducing mortality in L. donovani parasites at both the amastigote (intracellular) and promastigote (extracellular) stages, effectively targeting multiple life stages vital to the parasite's life cycle. Of these, S02587 showed particularly high selectivity, effectively killing the parasite while preserving host macrophage cell viability at the same dose, and minimizing risks of side effects on human cells. This selective toxicity underscores S02587's promise as a therapeutic candidate for leishmaniasis with fewer adverse effects. The findings establish HSK as a viable drug target for L. donovani and validate S02587 as a potent and selective inhibitor worth further exploration. By focusing on essential, parasite-specific metabolic pathways, this study advances a strategic approach for safer, more effective anti-leishmanial therapies, potentially leading to new treatment options with improved safety profiles.

Background and aims: Stanford type A aortic dissection (AD) is a degenerative aortic remodelling disease marked by an exceedingly high mortality without effective pharmacologic therapies. Smooth muscle cells (SMCs) lining tunica media adopt a range of states, and their transformation from contractile to synthetic phenotypes fundamentally triggers AD. However, the underlying pathomechanisms governing this population shift and subsequent AD, particularly at distinct disease temporal stages, remain elusive.

Methods: Ascending aortas from nine patients undergoing ascending aorta replacement and five individuals undergoing heart transplantation were subjected to single-cell RNA sequencing. The pathogenic targets governing the phenotypic switch of SMCs were identified by trajectory inference, functional scoring, single-cell regulatory network inference and clustering, regulon, and interactome analyses and were confirmed using human ascending aortas, primary SMCs, and a β-aminopropionitrile monofumarate-induced AD model.

Results: The transcriptional profiles of 93 397 cells revealed a dynamic temporal-specific phenotypic transition and a marked elevation of the activator protein-1 (AP-1) complex, actively enabling synthetic SMC expansion. Mechanistically, tumour necrosis factor signalling enhanced AP-1 transcriptional activity by dampening mitochondrial oxidative phosphorylation (OXPHOS). Targeting this axis with the OXPHOS enhancer coenzyme Q10 or AP-1-specific inhibitor T-5224 impedes phenotypic transition and aortic degeneration while improving survival by 42.88% (58.3%-83.3% for coenzyme Q10 treatment), 150.15% (33.3%-83.3% for 2-week T-5224), and 175.38% (33.3%-91.7% for 3-week T-5224) in the β-aminopropionitrile monofumarate-induced AD model.

Conclusions: This cross-sectional compendium of cellular atlas of human ascending aortas during AD progression provides previously unappreciated insights into a transcriptional programme permitting aortic degeneration, highlighting a translational proof of concept for an anti-remodelling intervention as an attractive strategy to manage temporal-specific AD by modulating the tumour necrosis factor-OXPHOS-AP-1 axis.

Introduction: Asthma has a strong allergic component, while Celiac Disease (CD) is an autoimmune disorder triggered by gluten ingestion in genetically predisposed individuals. This study investigates whether asthma and CD share genetic risk factors, offering insights into their biological interplay and overlapping pathways.

Methods: Pediatric patients diagnosed with asthma (n = 4848) or CD (n = 346) were compared to controls who had never been diagnosed of any allergic or autoimmune diseases (n = 1,963). Ninety-eight patients were diagnosed with both diseases. Asthma (PGS002727) and CD (PGS002067) polygenic risk scores (PRSs) were analyzed separately for females and males. All participants were of European ancestry and recruited by the Center for Applied Genomics (CAG) at the Children’s Hospital of Philadelphia (CHOP).

Results: We conducted a cross-examination, excluding patients diagnosed with both asthma and CD to avoid potential confounding effects. Key findings are as follows:

• Female Asthma Patients: Increased asthma PRS (P=2.37E-05) and CD PRS (P=0.018).
• Female CD Patients: Elevated asthma PRS (P=0.007) and CD PRS (P=1.04E-25).
• Male Asthma Patients: Increased asthma PRS (P=4.77E-04); CD PRS not significant (P=0.420).
• Male CD Patients: Higher asthma PRS (P=9.83E-06) and CD PRS (P=4.31E-16).
• Controls: Asthma and CD PRS correlated significantly (females: r=0.249, P=6.39E-15; males: r=0.232, P=8.56E-14).

Pathway PRS analyses using ImmuneSigDB C7 (5219 gene sets) showed 630 significantly altered pathway CD PRSs after Bonferroni correction. Pathway PRS associations for asthma were less pronounced. Two pathways identified in CD patients were also increased in asthma patients (P < 0.001 in both sexes): [THAKAR_PBMC_INACTIVATED_INFLUENZA_AGE_21_30YO_RESPONDERS_28DY_DN] and [GSE3039_CD4_TCELL_VS_B1_BCELL_UP].

Conclusions: Our findings highlight a shared genetic risk between asthma and CD. Two shared gene sets related to altered PBMC activity and CD4+ T cells were identified. These findings provide valuable insights into these overlapping mechanisms and identify targets for therapeutic interventions.

Thymic epithelial tumors (TETs), including thymomas and thymic carcinomas (TCs), are rare neoplasms comprising 0.2–1.5% of malignancies, characterized by heterogeneity in clinical behavior and molecular features. The Hippo signaling pathway, a key regulator of cellular proliferation and differentiation, has been implicated in TET progression, yet its precise role remains unclear. This study investigates the expression and regulation of core Hippo pathway components—YAP1, TAZ, SAV1, MOB1, LATS1, MST1, and TEAD4—in TETs through an immunohistochemical analysis of 77 samples. MOB1 and SAV1 were consistently expressed across all TET subtypes, suggesting stable Hippo pathway activity. MST1 showed low to moderate expression, with higher levels in TCs. LATS1 exhibited very strong expression in TCs and moderate to strong levels in other subtypes. TEAD4 displayed variable expression, being stronger in TCs. TAZ expression showed marked variability in A-B3 thymomas, while TAZ was nuclear negative in most B1-B3 thymomas but strongly expressed in the cytoplasm. In contrast, TAZ in TCs was predominantly cytoplasmically negative and showed little nuclear expression. YAP showed weak to moderate nuclear expression in A-B3 thymomas but was largely absent in TCs, with occasional cytoplasmic positivity. The findings indicate the differential regulation of the Hippo pathway across TET subtypes. The OFF state of the Hippo pathway in thymoma A may allow for "controlled" proliferation, supporting homeostasis without aggressive behavior. In contrast, TCs exhibit complex Hippo pathway dysregulation. The moderate to strong expression of TAZ, MST1, LATS1, and TEAD4 in TCs suggests a mechanism where TAZ substitutes for YAP, promoting pro-oncogenic TEAD4-mediated signaling, potentially driving uncontrolled proliferation and tumor progression. This highlights the potential involvement of other pathways or epigenetic factors in TCs. Further validation through Real-Time PCR and Next-Generation Sequencing is needed to confirm these findings, identify mutations, and uncover the mechanisms underlying TET tumorigenesis. These insights may inform therapeutic strategies targeting the Hippo pathway in TETs.

Type 2 diabetes mellitus (T2D) represents an indirect cause of mortality and a key factor in reduced quality of life due to its progressive impact on cardiometabolic health. Genetic predisposition is estimated to account for more than 50% of the risk of developing T2D, although most genetic variants remain unidentified. Identifying heterogeneous biomarkers, including genetic variants and their interaction with non-genetic factors, is crucial for advancing the understanding of T2D aetiology. This study aimed to identify genetic variants (SNPs) associated with T2D through Next-Generation Sequencing (NGS) and to investigate their interaction with environmental, clinical, and anthropometric factors in a cohort of the Spanish general population (DI@BET.ES dataset).

An exploratory analysis was conducted to evaluate the effect of prevalent genetic variants on T2D predisposition, as well as their interaction with environmental and clinical variables. To prioritize the identified variants, the SEQENS algorithm was employed to generate relevance rankings in stratified populations. Three different models were developed for the analysis: a genetic model, an environmental model, and a combined model. The performance of the prioritization models was evaluated using the concordance index of machine learning models predicting the risk of developing T2D as a function of age with the identified risk factors.

The combined model outperformed the individual models in variant prioritization and interaction analysis. The most relevant variables selected included fasting glucose, BMI, waist circumference, basal insulin, HDL and LDL cholesterol, triglycerides, consanguinity with first-degree relatives with T2D, body weight, WHR, monthly beer intake, nickel in particulate matter, and a specific genetic variant, rs78416608. This variant, located in the SCARA5 gene, is linked to immunity, apoptotic cell clearance, and lipid metabolism regulation, processes associated with T2D pathophysiology. These findings highlight the importance of an integrative approach to understanding T2D.

Introduction
The incidence of total knee and hip arthroplasty (TKA and THA) isrising due to demographic changes, leading to an increase in postoperative complications like prosthetic joint infection (PJI), aseptic failure, and chronic pain. The diagnosis of PJI is complicated and lacks a gold standard. Additionally, the diagnosis of chronic joint pain, which does not require revision surgery, is difficult to distinguish from PJI and aseptic loosening.

Methods
This study utilized blood plasma samples from the Prosthetic-Related-Infection and Pain (PRIS) study. Patients (N=98) were separated into several disease and outcome groups and samples were taken at up to four timepoints from baseline to follow-ups. Plasma samples were analyzed by utilizing quantitative label-free liquid chromatography–mass spectrometry. The statistical analysis was conducted in Perseus and Stata. Bioinformatic analyses were explored using the STRING database.

Results
The deep proteome analysis of the serum samples revealed more than 650 quantifiable proteins, of which multiple proteins were associated with clinical outcome measures like pain. A comprehensive analysis of the regulated proteins by, e.g., STRING network analysis of protein–protein interactions revealed that chronic joint pain is associated with pathways in coagulation, cholesterol metabolism and actin filament-based processes. Further, PJI and aseptic loosening were associated with pathways of the extracellular matrix. Subsequently, volcano plots identified significantly upregulated expression of seven proteins (Protein s100-A8/9, C-reactive protein, Serum amyloid A-1/2, Histone H2A type 1-H, and Von Wildebrand factor) in the PJI group and four proteins (Kallistatin, Haemoglobin subunit beta, 4-Aminobutyrate aminotransferase, mitochondrial, and Phosphatidylinositol-glycan-specific phospholipase D) in the chronic pain group.

Conclusion
The comprehensive panel of biomarkers identified in this study could possibly aid in the diagnosis of PJI and chronic joint pain following TKA or THA. Subsets of proteins allowed separation of the patient groups. These findings have the potential of enhancing patient outcomes and quality of life.