Search Results (2,241)

Background/Objectives: Human milk composition is shaped by gestational age at delivery and stage of lactation; however, proteomic differences between milk from mothers of preterm and term infants and their temporal patterns remain incompletely characterised. Methods: This prospective study enrolled 40 lactating mothers: 20 who delivered preterm infants (<32 weeks’ gestation) and 20 who delivered at term (37–42 weeks). Each provided milk samples during early lactation (first 10 days postpartum) and during later lactation (week five postpartum). Milk serum was analysed using quantitative data-independent acquisition mass spectrometry. Differential protein abundance was assessed separately at each time point; functional annotation was performed using Gene Ontology biological process analysis. Results: Eighty samples were analysed. On average, a total of 662 proteins were identified per sample, of which 169 were consistently quantified across all samples (1% FDR). During early lactation, 10 proteins differed significantly, with bidirectional changes and moderate effect sizes. At week five, 19 proteins were differentially abundant, predominantly higher in preterm samples. Immune-related proteins constituted the largest functional category at both stages. Immunoglobulin heavy constant gamma 4 remained consistently downregulated in preterm milk (1.6-fold lower abundance). Ferritin heavy chain (1.5) and HLA class II histocompatibility antigen gamma chain (1.8) were elevated only early, whereas calprotectin subunits S100A8 (5.6) and S100A9 (5.2) were markedly upregulated later. Conclusions: Proteomic differences vary across lactation stages, highlighting lactation stage as an essential contextual variable in comparative milk proteomics. Full article

Background: Sepsis is one of the leading causes of early death after a liver transplant, with a frequency of up to 45% and a high death rate of 50% in more severe forms. Standard diagnostic and therapeutic algorithms are often not applicable to this specific population, where immunosuppression, reperfusion injury, and systemic inflammation overlap and generate a clinical picture that is significantly different from sepsis in immunocompetent patients. Methods: This paper analyzes the available literature and clinical experiences of characteristic immune and hemodynamic profiles of sepsis after liver transplants. Biomarkers (IL-6, IL-10, HLA-DR, lactate, and IgM) are discussed as tools for assessing immune status and guiding timely interventions, including the early application of continuous renal replacement therapy (CRRT) and the selective use of IgM-enriched immunoglobulins. Results: Sepsis after liver transplantation frequently unfolds in two phases, an initial hyper-inflammatory response driven by cytokine release and reperfusion injury and a second phase of secondary immunoparalysis characterized by reduced HLA-DR expression and increased anti-inflammatory signaling. The immunometabolic shift appears to influence the clinical course and may inform therapeutic decision-making. The immunoparalysis phase is accompanied by mitochondrial dysfunction and impaired vascular reactivity. This type of mechanism contributes to hemodynamic instability and a reduced response to standard therapy. Individualized monitoring and early use of hemofiltration and immunomodulatory measures can improve results in carefully selected patients. Conclusions: In this setting, an individualized immunometabolic approach may complement standard sepsis management in liver transplant recipients. The introduction of biomarkers of immune function into routine practice and the recognition of early signs of exhaustion of the immune response can assist in timely therapeutic decision-making and improve survival. Full article

Atherosclerosis is a complex vascular disorder driven by oxidative stress, inflammation, and platelet activation. Agents capable of targeting multiple atherogenic pathways may provide improved therapeutic benefits. In this study, we evaluated the anti-atherogenic effects of chrysotoxine, a bibenzyl compound isolated from Dendrobium pulchellum, using in vitro models relevant to atherogenesis. Chrysotoxine significantly suppressed hemin-induced LDL oxidation by reducing lipid peroxidation and apolipoprotein modification. In an endothelial–monocyte co-culture model, chrysotoxine markedly attenuated lipopolysaccharide-induced monocyte adhesion, indicating inhibition of endothelial inflammatory activation. Chrysotoxine also inhibited platelet aggregation induced by arachidonic acid, ADP, and collagen in a concentration-dependent manner, with the strongest effects observed against arachidonic acid–mediated responses, suggesting modulation of the thromboxane pathway. Molecular docking analyses and cyclooxygenase activity assays further indicated that chrysotoxine may interact with both COX-1 and COX-2, exhibiting inhibitory activity in the low micromolar range. Collectively, these findings demonstrate that chrysotoxine modulates multiple key processes involved in atherogenesis, including oxidative LDL modification, vascular inflammation, and platelet activation. Although further in vivo studies are required, chrysotoxine represents a promising plant-derived candidate for the development of multi-target strategies against atherosclerotic disease. Full article

Background: Behçet’s disease (BD) is a systemic inflammatory disorder marked by recurrent mucocutaneous and ocular manifestations, predominantly affecting populations along the historic Silk Road. Genetic susceptibility, especially involving HLA-B*51, is well established. Spondyloarthritis (SpA) shares immunogenetic and clinical overlaps with BD, notably through associations with HLA class I alleles, particularly HLA-B*27. However, extended HLA-B allele profiling in these conditions remains limited. This study aimed to investigate the extended distribution of HLA-B alleles in patients presenting with clinical features suggestive of BD or SpA and to compare their clinical and laboratory profiles. Methods: In a prospective observational study at a Bulgarian rheumatology center, 120 patients with suspected BD or SpA were enrolled between January 2023 and June 2025. Diagnoses were confirmed using International Criteria for Behçet’s Disease (ICBD) and ASAS criteria for SpA. Comprehensive clinical evaluations, laboratory assessments including HLA-B typing by Sanger sequencing, and inflammatory markers were collected and analyzed. Results: Of the cohort, 15 patients (12.5%) were diagnosed with BD and 30 (25%) with SpA. HLA-B*51 was predominantly associated with BD, while HLA-B*27 and its heterozygous combinations were prevalent in SpA patients. Suspected BD patients exhibited significantly higher levels of inflammatory markers (CRP, ESR) and characteristic clinical features such as oral/genital ulcers and uveitis compared to non-BD patients. Suspected SpA patients showed longer disease duration, increased NSAID use and higher prevalence of enthesitis, psoriasis and peripheral arthritis compared to non-SpA patients. Conclusions: This study confirms the strong associations of HLA-B*51 with Behçet’s disease and HLA-B*27 with spondyloarthritis while revealing additional heterozygous and less common alleles that suggest a broader genetic influence. These findings highlight the genetic diversity and clinical heterogeneity of BD and SpA, supporting the use of extended HLA typing to improve the diagnosis and understanding of these diseases. Full article

Allopurinol, the most used urate-lowering drug for the treatment of gout, is associated with rare but life-threatening severe cutaneous adverse reactions (SCARs) such as Stevens–Johnson Syndrome (SJS), Toxic Epidermal Necrolysis (TEN), and Drug Reaction with Eosinophilia and Systemic Symptoms (DRESS) syndrome, but not Acute Generalised Exanthematous Pustulosis (AGEP). They are characterised by severe skin and systemic involvement and are associated with substantial morbidity and a high risk of mortality. This narrative review summarises evidence on the clinical presentation, epidemiology, risk factors, and preventive strategies for allopurinol-induced SCARs. Key risk factors include the presence of the HLA-B*58:01 allele, renal impairment, older age, female sex, heart disease, higher starting doses of allopurinol, and certain ethnicities, e.g., South Asian, Han Chinese, and African populations likely due to the higher prevalence of the HLA-B*58:01 allele. Risk mitigation strategies include genetic testing for HLA-B*58:01 in high-risk ethnic groups and avoiding allopurinol in those that are positive for the HLA-B*58:01 allele, starting allopurinol at a low-dose (e.g., 50–100 mg/day) and up-titrating it gradually at 4-week intervals, and avoiding high-dose allopurinol in those with risk factors (e.g., chronic kidney disease stage ≥3). In addition, risk stratification using prediction tools may enable a safer use of allopurinol. Full article

Anti-β2GPI/HLA-DR antibody, chronic endometritis (CE), and endometrial dysbiosis are likely to be associated with the etiologies of recurrent pregnancy loss (RPL). This prospective cohort study aimed to investigate these new risk factors together with conventional causes for RPL, and to evaluate pregnancy outcomes in women individually treated. A total of 87 women with RPL underwent conventional assessment together with anti-β2GPI/HLA-DR antibody measurements, CD138 immunohistochemistry for CE, and 16S rRNA sequence analysis for endometrial microbiome. Women with anti-β2GPI/HLA-DR antibody, CE, and endometrial dysbiosis received low-dose aspirin and heparin, antibiotics, and probiotics, respectively. Pregnancy outcomes of the participants were assessed. Anti-β2GPI/HLA-DR antibody, CE, non-Lactobacillus-dominant microbiome (NLDM)-1 (Lactobacillus + Bifidobacterium < 80%), and NLDM-2 (Lactobacillus without iners + Bifidobacterium < 80%) were detected in 16 (18.4%), 22 (25.3%), 27 (31.0%), and 46 (52.8%) women, respectively. Based on conventional assessment, 65.5% of women with RPL were classified as unexplained etiology; however, the percentage reduced to 16.1% when these new tests were assessed together. All 9 pregnancies with anti-β2GPI/HLA-DR antibody, 13 (92.9%) of 14 pregnancies with CE, and 24 (92.3%) of 26 pregnancies with NLDM-2 resulted in live birth. Assessment of these new tests may be clinically useful for reducing the proportion of unexplained RPL, and for providing high live birth rates if women receive relevant treatments. Full article

Background/Objectives: Vaccinations are a cost-effective strategy to reduce morbidity and mortality from infectious diseases, particularly in individuals with diabetes mellitus (DM). Despite recommendations for routine vaccination, uptake among adults with DM remains suboptimal. This study evaluates the effectiveness of interventions aimed at increasing Herpes Zoster (HZ) vaccine coverage in diabetic patients. Methods: This study included diabetic patients residing in the Romagna Local Health Authority (1.2 million inhabitants). Two complementary strategies were implemented: patient-oriented interventions (reminders/recalls) and provider-focused actions to engage general practitioners and diabetologists. Vaccination coverage in December 2023 was compared with December 2024, stratified by age, sex, and nationality. Primary outcomes were changes in coverage and intervention impact. Multivariate logistic regression assessed sociodemographic determinants of adherence. Results: As of 31 December 2023, 5182 diabetic patients had received at least one dose of Herpes Zoster vaccine (7.4% coverage). In 2024, overall coverage (11.1%) increased across all sociodemographic groups. Coverage rose to 12.8% in males and 9.0% in females and to 11.7% among Italian citizens, while foreign citizens showed lower absolute levels but a greater proportional increase. By age, individuals born after 1958 exhibited the largest relative growth, whereas those born in 1952–1958 reached the highest absolute coverage in 2024. Male sex, the 1952–1958 birth cohort, and Italian citizenship were associated with a higher likelihood of vaccination in both years. Conclusions: Our findings show a positive trend in Herpes Zoster vaccination among diabetic patients in Romagna HLA, driven by patient reminders, provider engagement, and age-targeted campaigns, while persistent sociodemographic disparities underscore the need for tailored strategies to optimize coverage. Full article

Background and Aims: Sepsis drives mortality in cirrhosis, yet the gut antimicrobial resistance (AMR) landscape remains unmapped in high-burden settings like India. This study aimed to integrate shotgun metagenomics with deep immunophenotyping to define the gut–immune–resistome axis and correlate specific microbial and genetic signatures with clinical outcomes in decompensated cirrhosis. Methods: We analysed 78 hospitalized patients with cirrhosis using stool shotgun metagenomics, multiplex cytokine arrays, and flow cytometry. The microbiome and resistome (AMR genes) were mapped and correlated with disease severity, immune function (monocyte HLA-DR, neutrophil CD64), and clinical endpoints including mortality. Results: Disease severity was characterized by a “Gram-negative bloom” (Klebsiella) alongside pathogenic Enterococcus expansion and novel markers: Clostridium sp. C5-48 (severe decompensation) and Sutterella (ascites). A specific, dense resistome predicted adverse outcomes; the quinolone-resistance gene QnrB4 correlated with mortality and immune paralysis, while the carbapenemase OXA-833 gene was linked to gastrointestinal bleeding. Notably, the commensal Ligilactobacillus salivarius was associated with systemic inflammatory cytokines. Conclusions: This study reveals a “pathogenic ecosystem” in Indian decompensated cirrhosis where the resistome is intrinsically linked to host immune failure. The identification of specific prognostic markers (QnrB4, OXA-833) and inflammatory associations with L. salivarius challenges generic probiotic use and underscores the urgent need for precision, resistome-targeted therapies. Full article

Background: Early excision and autologous split-thickness skin grafting are the cornerstone of surgical management in severe burn injuries. In patients with extremely extensive deep burns, the lack of available donor sites represents a major life-threatening limitation. In the exceptional situation of monozygotic twins, skin isografting offers a unique solution by providing immunologically compatible skin without the risk of rejection. Case report: We report the case of a 33-year-old man who sustained flame burns involving 95% of his total body surface area, resulting in an extremely poor initial prognosis (ABSI 14, UBS 245). After early resuscitation and staged surgical excisions, the absence of sufficient autologous donor sites precluded definitive coverage using conventional techniques. On day 3, the existence of a monozygotic twin brother was identified. HLA genotyping confirmed complete identity, and skin donation was authorized by an independent ethics committee. Methods: Definitive wound coverage was achieved using staged split-thickness skin isografts harvested from the donor twin. Ultra-thin grafts (<0.2 mm) were obtained in three procedures (days 7, 11, and 45), primarily from the scalp, thighs, and back, and applied following sequential excisions. Results: All grafts survived without immunological rejection. Donor-site morbidity was minimal, with rapid healing and only mild residual hypopigmentation. The patient was discharged to rehabilitation on day 145. At 5-year follow-up, wounds were fully healed, functional outcome was satisfactory, and quality of life was good, with return to work and full independence. Conclusions: Skin isografting from a monozygotic twin is a rare but effective salvage strategy for patients with massive deep burns when autologous donor sites are insufficient, provided that ethical, legal, and donor safety considerations are rigorously addressed. Full article

Influenza’s pandemic threat is driven by antigenic drift, which limits the efficacy of conventional vaccines. To address this challenge, we established a clinical serum-anchored computational design pipeline for a broad-spectrum multi-epitope mRNA vaccine (MEMV), bridging the gap between pure in silico design and clinical applicability. Using 36 longitudinal sera (d0/d28/d365) from 12 well-characterized human cohorts (6 vaccine recipients and 6 influenza patients) and high-density antibody-peptide microarrays, we empirically identified 12 immunodominant B-cell linear epitopes from the nucleoprotein (NP) of influenza A (H1N1/H3N2) and B viruses. These experimentally validated epitopes were combined with in silico-predicted conserved helper T-lymphocyte (HTL)/cytotoxic T-lymphocyte (CTL) epitopes (from NP/HA/NA) to construct MEMVs candidates, ensuring high antigenicity, non-toxicity, and 95.63% global HLA coverage. Molecular docking and 100 ns molecular dynamics (MD) simulations confirmed favorable conformational compatibility between MEMVs and Toll-like receptor 3 (TLR3) in silico immunization via C-ImmSim predicted robust B/T-cell responses and protective cytokine (IFN-γ/IL-10) production. Collectively, this pipeline shortens the preliminary design cycle for influenza vaccines, provides a standard epitope-combination strategy, and offers direct targets for follow-up in vitro/in vivo experiments. Full article

Background/Objectives: Chronic lung allograft dysfunction (CLAD) remains the leading cause of late graft failure after lung transplantation (LuTX). De novo donor-specific anti-HLA antibodies (dnDSA), especially HLA-DQ, have been implicated; we assessed associations between dnDSA (class and specificity) and CLAD after LuTX. Methods: We retrospectively analyzed all LuTX recipients transplanted from 2005–2018 at a single center (n = 585). dnDSA were measured by Luminex single-antigen bead assays (MFI > 1000) at 1, 3, 6, and 12 months and at least annually thereafter. CLAD was defined by ISHLT criteria; time-to-event comparisons used log-rank testing. Results: dnDSA developed in 151/585 recipients (25.8%), predominantly class II (129/585; 22.1%); class I dnDSA occurred in 52/585 (8.9%). CLAD occurred more frequently in dnDSA-positive than dnDSA-negative recipients (64/151; 42.4% vs. 109/434; 25.1%; p < 0.0001). Rejection-attributed death was higher in dnDSA-positive recipients (19/151; 11.3% vs. 25/434; 5.3%; p = 0.01). Both class I and class II dnDSA were associated with higher CLAD rates (log-rank p < 0.001 each). Locus-specific analyses identified HLA-DQ dnDSA as strongly associated with CLAD (p < 0.0001); DQ7 was the most frequent specificity (n = 44) and showed the strongest association (p < 0.0001). Conclusions: dnDSA after LuTX were associated with increased CLAD incidence and rejection-attributed mortality, with a prominent association for HLA-DQ—particularly DQ7. Full article

Background/Objectives: Non-steroidal anti-inflammatory drugs (NSAIDs) are widely prescribed to help alleviate pain and treat inflammation, but they are also recognized as common causes of severe cutaneous adverse reactions (SCARs), including Stevens–Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN). Despite their clinical importance, pharmacogenetic markers to predict individual susceptibility to NSAID-induced SJS/TEN remain insufficiently defined. This study investigated associations between HLA class I and II alleles, CYP2C9 polymorphisms, and NSAID-induced SJS/TEN in a Thai population. Methods: A total of 18 patients with NSAID-induced SJS/TEN and 54 NSAID-tolerant controls were enrolled. Genotype data from 183 unrelated Thai individuals without a history of drug allergy were included as a general population control group. Genotyping was performed for HLA class I and II alleles and the CYP2C9*3 variant. Results: HLA-DQB1*03:02 was significantly associated with NSAID-induced SJS/TEN (OR = 9.23, 95% CI = 2.19–38.83, p = 0.0024, Pc = 0.0312), particularly those triggered by piroxicam (OR = 13.71, 95% CI = 2.81–66.86, p = 0.0012, Pc = 0.0156). Additional associations were identified for HLA-B*56:01 and HLA-A*68:01 in the overall NSAID-induced SJS/TEN group. The subgroup analysis suggested that these alleles, along with HLA-DRB1*04:03, were associated with an increased risk of piroxicam-induced SJS/TEN. However, these associations did not remain statistically significant after Bonferroni’s correction. No significant association was identified for CYP2C9*3. Conclusions: This study identified specific HLA alleles, particularly HLA-DQB1*03:02, as candidate pharmacogenetic risk factors for NSAID-induced SJS/TEN in a Thai population, especially in piroxicam-associated cases. However, these associations should be considered exploratory. Larger, multicenter, multi-ethnic studies are required to validate these findings and clarify their potential clinical utility. Full article

Background: Nonalcoholic fatty liver disease (NAFLD) is common in rheumatoid arthritis (RA), and methotrexate (MTX) use raises concern about hepatotoxicity. We evaluated whether HLA-DRB1, PNPLA3, SLCO1B1, and MTHFR variants are associated with NAFLD, liver fibrosis, or MTX toxicity/pharmacokinetics in RA, after accounting for clinical covariates. Methods: In a cross-sectional cohort of 159 patients with RA, NAFLD, and fibrosis were assessed by FibroScan (CAP ≥ 275 dB/m; LSM > 8 kPa). We compared baseline characteristics by NAFLD status and fitted multivariable models for NAFLD, fibrosis, ALT elevation, and MTX toxicity; MTX pharmacokinetics were analyzed in 111 MTX-treated patients. Multiple testing was controlled using the Benjamini–Hochberg method. Results: The prevalence of NAFLD was 36%, and that of fibrosis was 11%. NAFLD patients had higher CAP and LSM, and markedly greater adiposity indices (body weight, BMI, waist and hip circumference, WC). BMI and WC were independently associated with NAFLD (BMI OR 1.27 per kg/m², 95% CI 1.16–1.40; WC OR 1.06 per cm, 95% CI 1.01–1.12). No HLA-DRB1, PNPLA3, SLCO1B1, or MTHFR variant showed an association that survived multiple-comparison correction. Among MTX users, 21/111 (19%) experienced toxicity. SLCO1B1 and MTHFR variants did not influence MTX pharmacokinetics; age was associated with lower dose-normalized MTX exposure, and cumulative dose was positively associated with exposure. Conclusions: In RA, adiposity—not the tested candidate pharmacogenes—drives NAFLD risk, and SLCO1B1/MTHFR variants do not support MTX dose adjustment. The findings emphasize routine clinical risk factors over single-gene testing for NAFLD and MTX hepatotoxicity in this setting. Full article

Introduction: Chronic lymphocytic leukemia (CLL) is a common adult leukemia often treated with fludarabine, cyclophosphamide, and rituximab (FCR). While effective, FCR can lead to therapy-related myeloid neoplasms (t-MN), including aggressive therapy-related acute myeloid leukemia (t-AML). Stem cell transplantation offers the best chance for long-term remission in these cases. Here, we report a rare case of t-AML with plasmacytoid dendritic cells (pDC-AML) developing after FCR treatment for CLL that was successfully treated with haplotransplantation. Case Presentation: A 57-year-old woman with CLL-B was treated with six cycles of FCR, achieving a complete response. Six years later, at age 63, she developed t-AML with a rare morphophenotypic subtype: acute myelomonocytic leukemia with plasmacytoid dendritic cells (pDC-AML) and monosomy 8. Diagnostic challenges included distinguishing this subtype from blastic plasmacytoid dendritic cell neoplasm (BPDCN). She was treated with high-dose cytarabine followed by haploidentical stem cell transplantation from her son. Haploidentical transplantation was prioritized due to the urgent clinical need in a patient with high-risk acute leukemia (therapy-related leukemia secondary to prior chemoimmunotherapy and failure to achieve complete remission following the standard 3 + 7 induction protocol). In this critical setting, the patient’s son was immediately available as an HLA-haploidentical donor. Prior to the performance of the haploidentical stem cell transplant from her son, no HLA-matched unrelated donor (MUD) could be identified. Another viable alternative would have been the utilization of umbilical cord blood-derived stem cells harvested from the patient’s twin granddaughters. She was closely monitored post-transplant for potential complications, including graft-versus-host disease (GVHD), post-transplant lymphoproliferative disorder, and thyroid dysfunction, all of which were ruled out during follow-up. The patient remains in complete remission 15 years after her initial CLL diagnosis and 8 years after the t-AML diagnosis and haplotransplantation. Notably, no residual CLL clone was detected at the time of t-AML development, and a benign polyclonal lymphocytosis observed between 2018 and 2020 spontaneously resolved without intervention. Conclusions: This case illustrates the potential for long-term survival in high-risk patients with therapy-related AML developed after cytotoxic treatment for lymphoid malignancies. Haplotransplantation from a semi-identical Human Leukocyte Antigen (HLA) donor proved to be a viable and effective treatment option despite the patient’s age and dual hematologic malignancies. Full article

Background/Objectives: Staphylococcus aureus virulence is tightly regulated by the agr (accessory gene regulator) quorum-sensing system. Targeting AgrC, the histidine kinase receptor that serves as a core regulator of agr signaling, represents a promising antivirulence strategy that circumvents conventional bactericidal pressure. Methods: In this study, structure-based virtual screening using AutoDock Vina was performed, followed by molecular dynamics simulations, to identify potent analogs of known AgrC inhibitors. Results: A cyclo[Ala-Phe-OLeu-Phe-D-Leu] exhibiting high binding affinity and stable receptor interaction was selected for further evaluation. Antimicrobial susceptibility testing confirmed that the compound did not inhibit bacterial growth. However, at a concentration of 16 µg/mL, it significantly inhibited hemolytic activity with high reproducibility, and the inhibition rate reached 77.60%. Quantitative reverse transcription PCR (RT-qPCR) demonstrated that the compound decreased some key AgrC-mediated genes, including agrC, agrA, saeS, hla, spa, fnbA, and lukS. Conclusions: These findings identify a promising cyclic pentapeptide inhibitor of AgrC that effectively attenuates S. aureus virulence without exerting bactericidal pressure. This work provides a valuable lead compound and offers novel insights for the development of advanced, safe, and effective antivirulence therapeutics. Full article