Anti-topoisomerase I (anti-Scl-70) antibodies are strongly associated with systemic sclerosis and fibrotic organ involvement. However, their significance in ANA-positive individuals without established systemic sclerosis remains unclear. We investigated whether isolated anti-Scl-70 positivity is associated with subclinical cardiac involvement. This retrospective single-center study included 486 ANA-positive individuals identified between 2016 and 2026. Patients were classified as anti-Scl-70-positive (n = 242) or anti-Scl-70-negative (n = 244). Individuals with systemic sclerosis, other rheumatologic diseases, cardiovascular disease (except hypertension), malignancy, or advanced renal/hepatic disease were excluded. All anti-Scl-70-positive patients remained free of systemic sclerosis during at least 12 months of follow-up. Electrocardiographic Diastolic Index-1 (EDI-1) and EDI-2 were calculated from standard 12-lead ECGs. Demographic characteristics, inflammatory markers, ANA titers, and most ENA antibodies were similar between groups. Anti-Ro52 positivity was more frequent in anti-Scl-70-positive individuals (p = 0.004). EDI-1 was significantly lower, whereas EDI-2 was significantly higher in the anti-Scl-70-positive group (both p < 0.001). Among patients with available echocardiographic data, anti-Scl-70-positive individuals demonstrated higher E/e' ratios (p = 0.004) and lower LV-GLS values (p = 0.046). EDI-1 showed a strong correlation with E/e' (rs = -0.898, p < 0.001). In multivariable analysis, anti-Scl-70 positivity was the only independent predictor of electrocardiographic diastolic dysfunction (p < 0.001). Isolated anti-Scl-70 positivity is associated with impaired electrocardiographic markers of diastolic function in ANA-positive individuals without overt systemic sclerosis. These findings suggest that isolated anti-Scl-70 positivity is associated with early subclinical myocardial involvement and support further investigation of ECG-derived diastolic indices as potential screening tools in this population.
We report the case of an 11-year-old female who presented with a progressively enlarging cystic neck swelling. CT demonstrated a multiloculated cystic lesion involving the right cervical soft tissue planes, suggestive of a cystic lymphangioma. There was no significant family history or clinical evidence of neurofibromatosis type 1 (NF1) at presentation. Surgical excision was performed, and histopathological examination with S100 immunohistochemistry confirmed the diagnosis of plexiform neurofibroma. At the one-year follow-up, the patient developed multiple café-au-lait macules and Lisch nodules of the iris, fulfilling the diagnostic criteria for NF1. The delayed appearance of these characteristic features established the diagnosis of evolving sporadic NF1. This case highlights that an isolated plexiform neurofibroma may be the initial manifestation of NF1 and can clinically and radiologically mimic other cervical soft tissue lesions. Careful long-term clinical, dermatological, and ophthalmological follow-up is essential for the timely recognition of additional diagnostic features and appropriate surveillance.
Neonatal isolated submandibular suppurative sialadenitis (NISSS) is a rare condition. We present a case of NISSS caused by Staphylococcus aureus (S. aureus) in premature triplet girls born at 28 + 1 weeks of gestation. Mother's milk culture tested positive for S. aureus, which is presumed to be the potential contributing factor of the infection. The first symptoms appeared in one of the girls at 31 days of age, followed by her sisters at 34 and 36 days of age, respectively. A search of the PubMed/Medline, ScienceDirect and Web of Science databases revealed that this is the first reported case of NISSS in triplets. We defined risk factors like an immature immune system, prolonged orogastric feeding and antidiuretic therapy for developing NISSS in neonates.
Carbapenemase-producing Enterobacterales have disseminated globally, largely via highly transmissible plasmids. Their emergence in companion animals is of particular concern, indicating that clinically important carbapenem-resistance determinants have spread beyond healthcare settings. This study characterized a multidrug-resistant Escherichia coli isolate from a healthy dog in South Korea. Antimicrobial susceptibility was determined by standardized reference methods. Hybrid whole-genome assembly resolved chromosomal and plasmid architectures, and plasmid transferability was assessed by conjugation assays. Comparative genomic analysis based on nucleotide identity and alignment coverage evaluated relatedness to publicly available blaKPC-carrying IncX3 plasmids. The E. coli ST7854 isolate was resistant to carbapenems (imipenem MIC = 8 µg/mL), third-generation cephalosporins, fluoroquinolones, tetracycline, gentamicin, phenicols, and trimethoprim-sulfamethoxazole, but susceptible to amikacin and colistin. Assembly resolved a 4.77 Mb circular chromosome and seven plasmids. The ESBL gene blaCTX-M-27 was chromosomally integrated, whereas carbapenem resistance was conferred by blaKPC-2 on a 54,805 bp IncX3 plasmid. This IncX3-blaKPC-2 plasmid was transferred to E. coli J53 at a frequency of 5.91 × 10⁻⁴, whereas the large IncFIB multidrug resistance plasmid was not co-transferred. Comparative analysis revealed extensive structural similarity with Korean clinical IncX3 plasmids, whereas most representative international plasmids shared only the conserved IncX3 backbone. A commensal E. coli ST7854 isolate from a healthy companion dog carried a conjugative IncX3-blaKPC-2 plasmid and chromosomally integrated blaCTX-M-27. Similarity to Korean clinical plasmids suggests interconnected human-animal dissemination, and this asymptomatic carriage of mobile carbapenem-resistance determinants supports continued genomic surveillance of antimicrobial resistance in companion animals within a One Health framework.
Biofilm formation is a key virulence factor in Staphylococcus aureus, contributing to bacterial persistence, antimicrobial resistance, and chronic infections. This study aimed to investigate the presence of biofilm-associated genes (fib, fnbA, fnbB, clfA, and clfB) in S. aureus isolates from dogs in Ilam, Iran. From December 2022 to September 2023, 250 swab samples were collected from nasal, oral, and rectal sites of dogs, yielding 81 S. aureus isolates confirmed by PCR amplification of the nuc gene. The prevalence of biofilm-associated genes varied, with clfA, clfB, and fnbA detected in 98.80% of isolates, fib in 63.00%, and fnbB in 16.00%. Notably, fnbA, clfA, and clfB were present in all rectal isolates, while fnbB was absent in this group. The findings highlighted the widespread presence of biofilm-related genes in S. aureus from dogs, suggesting their potential role in colonization and zoonotic transmission. The high prevalence of adhesion-associated genes underscored the need for monitoring biofilm-forming S. aureus in companion animals to mitigate antimicrobial resistance and public health risks.
•B. clausii can cause bacteremia or sepsis in pediatric or immunocompromised patients.•In immunocompromised hosts, epithelial damage and adhesion favor bloodstream entry.•WGS revealed a close genomic relationship between both B. clausii isolates.•Multiple genomic methods consistently supported isolate relatedness.
Testicular tuberculosis is a rare form of extrapulmonary tuberculosis (EPTB). Due to the nonspecific nature of its clinical manifestations and imaging features, it is often misdiagnosed as a malignant testicular tumor, leading to unnecessary radical orchiectomy. Accurate preoperative diagnosis is essential for organ preservation and appropriate antituberculous therapy. A 59‑year‑old male patient was admitted with a 4‑day history of left testicular enlargement. Pre‑admission ultrasonography suggested "a solid mass in the left testicle, suspected seminoma." Physical examination revealed enlargement of the left testicle with moderate consistency. Scrotal ultrasound, contrast‑enhanced ultrasound, and enhanced CT all supported the diagnosis of testicular malignancy. The patient underwent left radical orchiectomy. However, postoperative pathology revealed extensive acute and chronic inflammatory cell infiltration, caseous necrosis, and granulomatous inflammation within the testicular tissue, consistent with tuberculous pathology. The final pathological diagnosis was "left testicular tuberculosis." Testicular tuberculosis is an important differential diagnosis for testicular tumors. Despite advances in modern imaging techniques, atypical presentation remains highly prone to misdiagnosis. Clinicians should maintain a high index of suspicion for testicular tuberculosis in patients with atypical clinical and imaging findings of testicular masses. When feasible, obtaining tissue samples for histopathological and molecular examination preoperatively or intraoperatively may be crucial in avoiding unnecessary orchiectomy and achieving organ preservation.
Multidrug-resistant Klebsiella aerogenes poses a significant clinical challenge due to its antimicrobial resistance and biofilm-forming capacity in chronic wounds and device-associated infections. Bacteriophage therapy offers a promising alternative to conventional antibiotics, though effective delivery systems remain critical for clinical translation. This study reports the isolation, genomic characterization, and alginate hydrogel-based formulation of a lytic bacteriophage, KA_SGEB_01, targeting multidrug-resistant K. aerogenes. The phage was isolated from hospital sewage using multidrug-resistant K. aerogenes as a host. Comprehensive characterizations included plaque morphology and host range determination, multiplicity of infection (MOI) optimization, one-step growth kinetics, antibiofilm activity assays, and environmental stability testing. Complete genome sequencing and annotation were performed. Phage-loaded alginate hydrogels were developed and evaluated for release kinetics, long-term viability, and antibiofilm efficacy. KA_SGEB_01 produced clear plaques (1 ± 0.1 mm) with halos indicative of depolymerase activity and exhibited strict host specificity. The optimal MOI was 0.01 with a 15-minute latent period and the phage remained stable between -20 °C and 50 °C; and across a pH range 6-9. Biofilm assays demonstrated around 60% Crystal-Violet-stained biomass reduction as well as significant biofilm disruption visualized by Scanning Electron Microscopy. Whole-genome sequencing revealed a 175,095 bp double-stranded DNA genome encoding 289 predicted coding sequences, classified within the family Straboviridae, genus Slopekvirus, and lacking any virulence or AMR genes. Encapsulation in alginate hydrogels preserved viability (>10⁹ PFU/mL) for 14 days and enabled sustained release (∼ 67% at 72 h), resulting in significant biofilm biomass inhibition. Collectively, KA_SGEB_01-loaded alginate hydrogels represent a promising platform for treating multidrug-resistant K. aerogenes in wound and device-associated infections caused by the bacterium.
To investigate the effects of young human red blood cell-derived extracellular vesicles (Y-RBCEVs) on fracture healing. Associations of RBC-related indicators with bone mineral density and mortality in fracture populations were analyzed using NHANES data (1999-2020). A murine femoral fracture model was established to investigate RBC transfusion effects. Y-RBCEVs and O-RBCEVs were isolated to examine their effects on osteogenic differentiation and BMM polarization in vitro. In vivo distribution was assessed by IVIS, and therapeutic efficacy was evaluated in fracture mice. Proteomic sequencing identified differentially expressed proteins between Y-RBCEVs and O-RBCEVs, and key protein function was investigated by transcriptomic analysis and in vitro validation. The observational study revealed associations between RBC count and bone mineral density with fracture populations mortality. RBC transfusion modulated the expression of osteogenic and M2 macrophage polarization-related genes. In vitro, RBCEVs were efficiently internalized by BMSCs and BMMs, with Y-RBCEVs enhancing osteogenic activity in BMSCs and promoting M2 macrophage polarization, thereby partially restoring osteogenesis impaired by inflammatory conditions. In vivo, Y-RBCEVs accumulated at fracture sites and enhanced bone regeneration without detectable toxicity, accompanied by increased OCN and CD206 expression. Proteomic analysis identified VCAN as one of the potential functional cargos. Mechanistically, Y-RBCEV-derived VCAN interacted with CD44 on macrophages and was associated with activation of the PI3K/AKT pathway, contributing to M2 polarization. Y-RBCEVs enhance fracture repair by modulating osteogenesis and macrophage polarization, with VCAN-associated CD44/PI3K/AKT signaling contributing to their osteo-immunomodulatory effects. These findings highlight RBCEVs as a potential therapeutic strategy for bone regeneration.
Perilunate dislocations (PLD) and perilunate fracture-dislocations (PLFD) are uncommon but severe carpal injuries. Although open or arthroscopic-assisted reduction with capsuloligamentous repair is recommended when feasible, access to suture anchors, appropriate nonabsorbable sutures, wrist arthroscopy, and advanced imaging may be inconsistent in resource-limited trauma settings. In such circumstances, temporary K-wire stabilization without formal ligament repair may represent a pragmatic stabilization strategy. We retrospectively reviewed 11 consecutive patients with acute PLD or PLFD treated between July 2019 and June 2024 in a single-center retrospective therapeutic case series (Level IV evidence). All patients underwent attempted fluoroscopic closed reduction followed, when necessary, by open reduction. Stabilization consisted of scapholunate and lunotriquetral pinning using two 1.6-mm K-wires, without formal ligament repair; associated fractures were fixed with K-wires as indicated. Outcomes included pain (VAS), QuickDASH, PRWE, Mayo Wrist Score (MWS), wrist range of motion, and grip strength. Radiographs assessed scapholunate distance (SLD) and scapholunate angle (SLA). Mean age was 36.9 years and 81.8% of patients were male. All injuries resulted from high-energy trauma (nine road-traffic accidents and two sports-related injuries). Five patients had PLD and six had PLFD. Closed reduction was successful in three cases; eight required open reduction (six dorsal and two combined dorsal-volar). Mean follow-up was 50.5 months. Mean VAS was 3. Mean QuickDASH was 30.4, PRWE 37.5, and MWS 78.5. Mean flexion-extension arc was 101.8° and mean grip strength was 75.2% of the contralateral side. Mean SLD was 3.0 mm and mean SLA was 61.8°. No pin-tract infection, K-wire migration, avascular necrosis, or radiographic osteoarthritis was observed at mid-term follow-up. In this small Level IV case series, isolated K-wire stabilization without formal ligament repair was associated with heterogeneous mid-term outcomes after acute PLD and fracture-dislocations. Partial functional recovery was observed in many patients, but residual scapholunate malalignment persisted in some cases. These findings are descriptive and hypothesis-generating and should be interpreted cautiously in the absence of a comparative cohort. They should not be viewed as evidence of equivalence to ligament-repair techniques.
Recurrence of posterior tibial translation (PTT) is one of the challenges of posterior cruciate ligament (PCL) reconstruction. Several studies have emphasized the importance of protecting the transplanted graft. However, the impact of such protective measures on clinical outcomes has rarely been investigated. The purpose of this study was to compare postoperative outcomes between conventional and modified rehabilitation (CR and MR, respectively) protocols after double-bundle PCL reconstruction. It was hypothesized that an MR protocol designed to protect the transplanted graft, combined with a dynamic PCL brace, would improve PTT and patient-reported outcomes (PROs) compared with CR. Cohort study; Level of evidence, 3. This study included 25 patients with isolated grade 2 or 3 PCL injuries who underwent double-bundle reconstruction using autogenous hamstring grafts between 2017 and 2022. Patients were categorized into 2 groups: the CR group (n = 14), which received CR without a dynamic PCL brace, and the MR group (n = 11), which received MR with a dynamic PCL brace. PTT was evaluated by gravity sag view radiographs preoperatively, immediately after surgery, and at 3 months, 6 months, 1 year, and 2 years after surgery. PROs were assessed using the Knee injury and Osteoarthritis Outcome Score (KOOS) at 2 years. PTT decreased from preoperative to immediate postoperative evaluation in both groups without a significant difference between the 2 groups (CR group: 9.9 ± 3.0 mm to -1.0 ± 1.9 mm; MR group: 9.6 ± 3.5 mm to -1.4 ± 0.5 mm). At 3 months, 6 months, 1 year, and 2 years after surgery, the CR group demonstrated PTT of 3.7 ± 1.5, 4.0 ± 1.7, 4.4 ± 1.6, and 4.7 ± 1.7 mm, respectively, whereas the MR group demonstrated significantly smaller values of 0.8 ± 1.9, 1.9 ± 1.6, 2.6 ± 2.1, and 2.6 ± 2.2 mm, respectively (P < .05). Residual posterior sagging (PTT >5 mm) occurred in 29% of the CR group and 9% of the MR group (P = .34). No major complications were observed. The KOOS Sport and Recreation subscale score tended to be higher in the MR group. MR with a dynamic PCL brace to protect the graft significantly improved tibial position after PCL reconstruction compared with CR, without increasing the risk of complications.
Soy protein isolate (SPI)-polysaccharide-stabilized high internal phase emulsions (HIPEs) are promising fat alternatives for meat analogs. However, the distinct contributions of polysaccharide charge and viscosity to emulsion formation, stabilization and meat analog quality remain unclear. This study systematically investigated five polysaccharides with different charges and viscosities, namely cationic chitosan (CS), neutral locust bean gum (LBG) and guar gum (GG), and anionic xanthan gum (XG) and high methoxyl pectin (HP), in modulating SPI structure, HIPE performance, and plant-based patty quality. Results showed that polysaccharide charge dominated SPI-polysaccharide interactions and dictated SPI conformational remodeling. Neutral polysaccharides interacted with SPI through hydrogen bonding/hydrophobic interactions, significantly increasing the β-sheet content (GG: from 35.91% to 43.23%), and achieving the smallest initial droplet size through a high interfacial adsorption rate (GG: 6.81 μm). During emulsion stabilization and flavor retention, viscosity was the predominant factor. High-viscosity XG and GG excelled in inhibiting droplet aggregation, reducing gravitational separation, and maximizing volatile flavor retention under both heating and storage. At the application stage, viscosity predominantly influenced the hardness of patties. This study reveals the division of labor between polysaccharide charge and viscosity across different performance dimensions of HIPEs, guiding polysaccharide selection for tailoring HIPEs in meat analogs.
Actinoplanes teichomyceticus is a well-established producer of bioactive secondary metabolites, including the glycopeptide antibiotic teicoplanin. Although its antibiotic biosynthetic capacity has been extensively investigated, its siderophore diversity and any additional biological functions of these iron-chelating metabolites remain comparatively underexplored. We identified a reproducibly bioactive, teicoplanin-independent fraction that inhibited Bacillus spizizenii. Molecular networking applied to this fraction identified hydroxamate ferrioxamine and desferrioxamine-type siderophores as the dominant metabolites, including acylated analogs detected as Al3+- and Fe3+-chelated species. Robust siderophore secretion was confirmed by the CAS assay. Notably, siderophore-enriched fractions exhibited selective antibacterial activity against Gram-positive bacteria, with minimum inhibitory concentrations of approximately 16 µg/mL against B. spizizenii and partial inhibition of Staphylococcus aureus, while no activity was observed against Escherichia coli. Synthetic C7 and C9 acyl-desferrioxamine analogs showed enhanced antibacterial activity upon Al3+ chelation, indicating a metal-dependent bioactivity. These findings reveal an unexpected antibacterial role for ferrioxamine-type siderophores produced by A. teichomyceticus, extending their function beyond iron acquisition, possibly through a "Trojan horse" (or "Trojan metal") mechanism.
Azo dyes represent one of the most persistent classes of industrial pollutants due to their structural stability and resistance to biodegradation. In this study, the azo dye removal mechanism of Bacillus proteolyticus was investigated using an integrated approach combining microbiological assays, FT-IR spectroscopy, molecular docking, and molecular dynamics simulations. The isolate (B. proteolyticus OA7), obtained from textile wastewater, was identified by 16S rRNA gene analysis. Decolorization experiments demonstrated a markedly higher removal efficiency for Acid Red 88 (69.74%) compared to Acid Blue 193 (41.01%), indicating substrate-dependent biodegradation behavior. Solid medium assays revealed distinct removal patterns, where Acid Red 88 underwent both biosorption and biodegradation, while Acid Blue 193 was primarily removed via biosorption. FT-IR analysis showed significant alterations in hydroxyl, lipid, protein (Amide I-II), and extracellular polymeric substance (EPS) functional groups following dye exposure, suggesting the involvement of both surface interactions and intracellular responses. The emergence of aromatic vibration bands (900-600 cm-1) in dye-treated samples supports the occurrence of azo bond transformation. Molecular docking analyses demonstrated strong binding affinities of azo dyes toward key redox enzymes, including azoreductase, OYE-like flavoprotein reductase, FMN-dependent NADPH-quinone reductase, and laccase, indicating their potential role in enzymatic degradation. Notably, Acid Red 88 exhibited favorable binding without hydrogen bond formation, highlighting the contribution of hydrophobic interactions. According to molecular simulations, ligand binding induced moderate conformational changes and local flexibility, but did not affect the global structural stability of the enzyme. Overall, the findings support a two-stage azo dye removal mechanism involving initial biosorption followed by enzymatic redox-mediated degradation. This study provides new mechanistic insights into the role of B. proteolyticus in azo dye bioremediation and highlights the influence of dye structure on bacterial response and removal efficiency.
To quantify the magnitude and trends the national and subnational burden of Alzheimer's disease and other dementias (ADOD) in Mexico from 1990 to 2023, analyzing patterns by sex and age and exploring their association with the Socio-Demographic Index (SDI) and the Healthcare Access and Quality Index (HAQI). A secondary ecological study was conducted using updated estimates from the Global Burden of Disease and Risk Factors Study (GBD) 2023. Prevalence, incidence, mortality, and disability-adjusted life years (DALYs) were examined. Temporal trends were assessed using joinpoint regression. Pearson correlation and linear regression were used to evaluate associations between DALYs rates and SDI and HAQI. Between 1990 and 2023, ADOD prevalence and incidence increased, despite significant declines in age-standardized prevalence and incidence rates. Females consistently experienced higher mortality and disability, with age-standardized DALYs rates 1.28 times those of males. The ADOD burden increased sharply with age, peaking among those aged 85 years and older, with premature mortality accounting for 63.0% of total DALYs. A significant increase in DALYs rates occurred during 2020-2023 after previous periods of gradual decline. DALYs rates were negatively correlated with both SDI and HAQI. ADOD disproportionately affect women, while higher modeled burden was observed in several states with lower socioeconomic development and weaker health system performance. The post-2020 increase represents an epidemiological signal warranting further investigation into excess mortality among people with dementia, healthcare disruption, social isolation, and changes in long-term care during the COVID-19 period. Strengthening early diagnosis, long-term care, and management of modifiable risk factors is essential to reduce the future burden and persistent regional and sex-based inequalities in ageing populations.
This study aimed to detect Coxiella burnetii, Chlamydia abortus, and Brucella species in the abomasal contents of aborted ruminant fetuses from the Central Anatolia region of Türkiye using PCR between 2020 and 2023. The abomasal contents of a total of 97 aborted fetuses from cattle, sheep, and goats with a history of abortion, collected between the years 2020 and 2023, were tested in this study. As a result of PCR analysis of 97 abomasal contents, four (4.10%; 95.00% confidence interval [CI]: 1.33 - 10.82) of them were C. abortus, including three sheep and one goat. Two (2.10%; 95.00% CI: 0.36 - 7.96) of them were C. burnetii, including one sheep and one cow. A total of 60 (61.90%; 95.00% CI: 51.40 - 71.37) samples from 47 cattle, nine sheep, and four goats were determined by Brucella genus-specific PCR. Following multiplex PCR analysis of the positive Brucella spp. samples, 39 (65.00%; 95.00% CI: 51.52 - 76.55) samples were identified as B. abortus, including two sheep, one goat, and 36 cattle. Additionally, 19 (31.70%; 95.00% CI: 20.60 - 45.09) isolates were identified as Brucella melitensis, including five sheep, two goats, and 12 cattle. In two sheep samples, both B. melitensis and C. abortus were identified from the same animals. In conclusion, Brucella spp. were the predominant abortion-causing pathogens, with C. abortus also contributing significantly. Effective control strategies under the One Health approach are essential to prevent the uncontrolled spread and inter-species transmission of these zoonotic agents in the region and country.
Artificial intelligence (AI) is gradually permeating every aspect of daily life. AI is now more than a mere tool; it significantly influences individual capabilities, professional values, social status, and our perspectives on the future. Amid these AI-driven changes, the concept of AI anxiety (AIA) has emerged to describe the associated emotional distress; however, its psychiatric significance has not yet been clearly defined. Presently, AIA can be conceptualized not as a distinct psychiatric diagnosis, but as a stress response capable of amplifying pre-existing vulnerabilities, psychiatric symptoms, maladaptive coping mechanisms, and functional impairments. This review categorizes AIA into five interrelated domains: anxiety regarding competence and adaptation, anxiety concerning occupational displacement and role loss, anxiety related to sociotechnical mistrust and loss of control, anxiety regarding identity and the human-machine boundary, and existential or catastrophic anxiety regarding the future. The clinical assessment of patients should evaluate specific AI-related experiences, perceived threats, behavioral responses, functional consequences, comorbid symptoms, and reality-testing abilities. Cases accompanied by excessive reliance on AI, social isolation, or the substitution of human support with AI interactions warrant increased attention. Therapeutic approaches include psychoeducation, cognitive and behavioral strategies, acceptance-based approaches, meaning-centered psychotherapy, AI literacy education, and efforts to restore human connections. Future studies should clarify the definition and scope of AIA and elucidate its course and treatment responsiveness through longitudinal and clinical studies using reliable assessment tools.
Traumatic brain injury (TBI) involves both primary and secondary pathological processes, including hemorrhage, ischemia, edema, and neuroinflammation. Although tissue-derived exosomes have emerged as important mediators of intercellular communication within local tissue microenvironments, their role in TBI-associated inflammatory responses remains incompletely understood. This study integrated brain tissue-derived exosomal proteomics with publicly available hippocampal transcriptomic data to investigate the potential association between tissue-derived exosomes and chemotaxis-related inflammatory responses after TBI. Brain tissue-derived exosomes were isolated, characterized, and subjected to label-free proteomic analysis. Public hippocampal transcriptomic data from GSE173975 were analyzed, and differentially expressed proteins (DEPs) and genes (DEGs) were identified from the exosomal proteomic and transcriptomic datasets, respectively. Functional enrichment, co-enrichment, and protein-protein interaction network analyses were performed, followed by quantitative real-time PCR, exosomal western blotting, and Transwell migration assays for validation and functional assessment. In total, 190 DEPs and 465 DEGs were identified. Co-enrichment analyses highlighted immune- and inflammation-related processes, particularly chemotaxis-related processes and chemokine signaling. LGALS3 and ITGB2 were increased at the exosomal protein level in brain tissue-derived exosomes from TBI rats and also showed increased hippocampal mRNA expression, whereas CCL2, CCL3, and CCR5 were upregulated at the hippocampal mRNA level. Functionally, TBI-derived exosomes enhanced BV2 microglial migration in vitro. Overall, these findings suggest that injury-associated brain tissue-derived exosomes may be associated with chemotaxis-related inflammatory and pro-migratory responses after TBI.
Xenotransplantation using genetically engineered pig organs offers a promising solution to the shortage of donor organs for life-saving transplantations. However, human-preformed antibodies against unknown pig xenoantigens remain a significant barrier to successful xenotransplantation. Current methods for characterizing these antibodies or xenoantigens are limited to cellular-level cross-match assays. In this study, we developed a novel approach to identify pig xenoantigens, including peptide and glycopeptide epitopes, that react with human-preformed antibodies. First, human-preformed antibodies against xenoantigens were enriched from plasma using immobilized pig kidney proteins. The enriched antibodies were then immobilized and used to isolate pig kidney proteins, peptides, and intact glycopeptides, followed by liquid chromatography-tandem mass spectrometry analysis. This dual-level approach identified 221 peptides corresponding to 153 proteins, with a significant enrichment of plasma membrane and extracellular proteins. Notably, 11 peptides were unique to pig sequences, suggesting their potential role in driving xenogeneic immune responses. Glycoproteomic analysis identified 122 intact glycopeptides, predominantly complex/hybrid glycoforms, and Neu5Gc-containing glycans. Our method effectively identifies peptides and intact glycopeptides reactive to human-preformed antibodies, providing critical insights for discovering xenoantigens. These findings could guide genetic engineering strategies and enhance recipient candidate screening for xenotransplantation, ultimately increasing the feasibility and success of xenogeneic organ transplantation.
To reduce potential inconsistencies in particle transport, temporal response, and microenvironment in conventional split-type aerosol absorption-scattering systems, we developed a dual-wavelength spatiotemporally synchronized measurement method. Two integrating spheres serve as scattered-light collection cavities and acoustic buffers and form a low-frequency differential Helmholtz resonator, while the central tube supports a high-frequency differential second-order longitudinal mode. The 450 and 532 nm lasers were modulated at 2572 and 169 Hz and synchronously demodulated using an FPGA-based digital lock-in amplifier. Absorption channels were calibrated with NO₂, and scattering channels with N₂/CO₂ Rayleigh scattering. At 1 s integration and a 3σ criterion, the 532 nm absorption and scattering limits of detection were 1.2 and 1.32 Mm⁻¹ . Water-mist, controlled-combustion, and field tests evaluated channel isolation, relative spectral response, and ambient-operation feasibility. The system simultaneously measures dual-wavelength absorption and scattering within a common volume through a single sampling path.