Immunity, Inflammation and Disease

This work aimed to establish a translationally viable, xeno-free, serum-free platform and protocol for the isolation and expansion of human salivary stem/progenitor cells (hS/PCs) suitable for regulatory qualification and future FDA-approved first-in-human autologous regenerative therapy trials for the treatment of hyposalivation disorders. Parotid gland specimens from non-cancerous regions/tissues were collected from consented surgical patients. Primary hS/PCs were isolated from tissue specimens, cultured in animal-component-free conditions, expanded to produce millions of cells, then enriched for CD44+ stem/progenitor cells by magnetic cell sorting. Normal epithelial purity was assessed using cytokeratins 5/14. Anti-CD133/PROM1 (cancer marker) and anti- fibroblast (clone TE-7) antibodies were used to demonstrate a lack of contaminating cells. Phenotype validation was performed by flow cytometry and immunocytochemistry on both CD44+ sorted and unsorted populations. Senescence-associated beta-galactosidase (SA-{beta}-gal) assays were performed across serial passages (P1-P6). Pluripotency was demonstrated by culture under conditions supporting lineage-specific differentiation. Primary hS/PCs demonstrated consistent expansion and epithelial morphology under serum-free conditions. CD44 expression remained high (>95%) throughout expansion, with negligible detection of CD133 or fibroblast markers, confirming epithelial purity and absence of tumorigenic or stromal contamination. Immunocytochemistry corroborated these expression profiles. SA-{beta}-gal staining revealed only a minor, passage-dependent increase (5-16%) in senescent cells from multiple donors, indicating retention of proliferative potential. Our defined, animal-free culture system supports stable expansion of pure low passage hS/PCs under conditions compatible with good manufacturing practice (GMP).

Aim: To investigate whether salivary immune cell profiling can serve as a non-invasive approach to monitor periodontal disease activity and therapeutic response by characterizing innate and adaptive immune cell dynamics in periodontitis. Materials and Methods: This longitudinal study included systemically healthy adults with periodontitis and healthy controls. Periodontal parameters (PPD, BOP, plaque/calculus, and radiographic bone loss) were recorded by calibrated examiners following established criteria. Stimulated saliva and gingival biopsies were collected before and 4-6 weeks after non-surgical periodontal therapy (NSPT), and from healthy controls. Multiparametric flow cytometry was used to characterize myeloid and lymphoid cell populations and polarization markers. Bacterial transcripts and host inflammatory markers were assessed by qRT-PCR. Statistical analyses were performed using one-way ANOVA. Results: Periodontitis subjects exhibited significantly elevated salivary bacterial transcripts, which decreased but did not normalize following NSPT. Both myeloid and lymphoid immune cell populations increased in periodontitis compared with healthy controls and declined after therapy. This was accompanied by a pronounced pro-inflammatory shift with elevated IFN-gamma-producing macrophages, dendritic cells, Th1/Th17 cells, and B cells, including the novel identification of IFN-gamma-producing B cells in saliva and mirrors the gingival immune cell profiles. In contrast, anti-inflammatory populations (IL-10-producing myeloid cells, Tr1 cells, and regulatory B cells) were reduced in disease and partially restored following NSPT. Conclusions: Salivary immunophenotyping non-invasively monitors PD activity and therapeutic response by capturing dynamic immune changes that reflect gingival signatures and track post-therapy resolution.

Unlike the conventional mature neutrophils, immature neutrophils have been investigated for their regenerative properties; however, their limited availability necessitates alternative generation strategies. Here, we used a combination of dimethylsulfoxide (DMSO) and 1,25-dihydroxyvitamin D3 (D3) to differentiate myeloid leukemia (HL-60) cells into immature neutrophil-like cells. Differentiated cells exhibited reduced cell size, loss of uniformity, decreased nuclear-to-cytoplasmic ratio, band-shaped nuclei, increased proportion of CD11b+CD14+ cells (indicative of immature neutrophils), decreased proportion of CD11b+CD16+ cells (indicative of mature neutrophils), higher levels of arginase 1, TGF{beta}1 (markers of immature neutrophils), and no expression of CD16, MRC1 (markers of mature neutrophils and M2 macrophages, respectively). Proteomic analysis revealed enrichment of proteins associated with immature neutrophils and wound healing. Functionally, these cells supported limbal stem cell growth and wound closure in vitro, indicating relevance for corneal regeneration. Administration of these cells to ex-vivo and in-vivo alkali-injured corneas, resulted in significant effect on promotion of wound healing, with epithelial regeneration and decreased fibrotic markers, proving that such cells hold promise for clinical translation as a therapeutic tool for tissue repair.

ObjectiveX-linked Hypophosphatemia is associated with dental complications, including spontaneous endodontic infections (abscesses) in non-carious teeth and severe periodontal loss. Previous studies have mainly focused on dentin Hypomineralization; however, the structural basis underlying periodontal tissue failure remains unclear. We aimed to investigate histoanatomical abnormalities in the dentin and periodontium of Hyp mice to clarify structural consequences of Phex deficiency in adult molars. MethodsWe performed detailed histological and scanning electron microscopy analyses on the molar regions of untreated adult Hyp mice and wild-type littermates, with particular attention to the structural integrity of the root and periodontal ligament. Additionally, odontoblast process morphology and periodontal attachment abnormalities were evaluated. ResultsHyp molars exhibited marked root abnormalities, including radicular shunt-like defects and disorganized odontoblast processes, particularly in furcation and radicular dentin. Periodontal attachment showed characteristic asymmetry: detachment from the cementum surface was frequently observed, whereas attachment to the alveolar bone surface was relatively preserved. These changes were accompanied by thinning and discontinuity of Sharpeys fibers and increased vascularity in the periodontal ligament. ConclusionsThese findings provide a histoanatomical framework for understanding refractory dental complications in X-linked hypophosphatemia and support the importance of intervention during root development.

The 5th Edition of the World Health Organization Classification of Haematolymphoid Neoplasms (WHO5th) and the 2022 International Consensus Classification (ICC) both recognize myelodysplasia-related acute myeloid leukemia (AML-MR) as a diagnostic entity increasingly defined by integrated genomic data. Although largely concordant, the two classifications differ in various ways that should be resolved to achieve future harmonization. To address the areas of uncertainty, we retrospectively analyzed 615 newly diagnosed AML cases from adult patients treated at two large cancer centers. We demonstrate that AML-MR, whether defined by gene mutations (MR-GM) or cytogenetic abnormalities (MR-CGA), constitutes a prognostically distinct group with inferior outcome compared to most AML subtypes, second only to TP53-mutated or EVI1-rearranged AML. Isolated RUNX1 mutations were not associated with antecedent myeloid neoplasia. Neither the number of mutated MR genes nor their variant allele frequency independently impacted outcomes. Trisomy 8 and del(20q) did not confer inferior outcomes and may warrant exclusion from MR-CGA. Complex karyotype without TP53 mutations did not worsen outcomes within AML-MR and may be considered equivalent to other MR-CGA. The adverse prognosis of AML-MR appeared to be at least partly driven by ASXL1 and/or EZH2 mutations. These findings provide evidence toward a unified schema across the WHO5th and ICC.

Background Alveolar bone assessment in periodontal practice relies on radiography and clinical probing, both of which have well-documented limitations in precision. Intraoral high-frequency ultrasonography (US) offers a radiation-free alternative with potential for sub-millimeter resolution, the validity and precision for detecting minute osseous changes have not been established. The purpose of this study was to evaluate the concurrent validity and measurement precision of intraoral US for detecting alveolar bone-level changes in patients undergoing crown lengthening and osseous surgery, thereby enabling its translation to monitor osseous changes in patients with periodontitis. Methods Ten patients (28 tooth sites) undergoing crown lengthening or osseous surgery at a USC Advanced Grad Perio clinic were enrolled in this prospective observational study. Distance from the cementoenamel junction (CEJ) to the Alveolar bone crest (ABC) was measured at pre- and post-operative time points using a 40 MHz handheld intraoral US transducer and, intraoperatively, by standardized clinical photography. Agreement was assessed by Pearson correlation and Bland-Altman analysis. Measurement precision was quantified using the standard error of measurement (SEM) and minimum detectable change (MDC). Results Preoperative agreement between methods was excellent (r = 0.977; Bland-Altman bias = -0.009 mm; 95% limits of agreement [LoA]: +-0.40 mm). Post-operative correlation remained strong (r = 0.912; bias = 0.123 mm; LoA: -0.85 to +1.10 mm). Both methods detected statistically significant post-surgical increases in the ABC-to-CEJ distance (p < 0.001), as anticipated. US demonstrated substantially superior precision: preoperative SEM 0.058 mm with US versus 0.128 mm clinically, yielding MDC values of 0.160 mm (US) versus 0.354 mm (clinical), providing a 2.2-fold precision advantage. Conclusions Intraoral US demonstrated strong concurrent validity with clinical photography and a reproducible precision advantage in detecting alveolar bone-level changes in patients with periodontitis. These findings support its clinical utility as a radiation-free, high-sensitivity bone monitoring tool. Larger longitudinal studies with CBCT validation are warranted.

Rationale: Infections due to Extended-spectrum {beta}-lactamases-producing Enterobacterales (ESBL-PE) require empirical treatment with carbapenems. ESBL-PE carriage is considered as a risk factor for ESBL-PE involvement during ICU infection. Our aim was to determine factors that may predict the actual involvement of ESBL-PE. Methods: A two-periods bicentric ambispective study including ICU ESBL-PE carriers patients from April 2011 to January 2019. All ESBL-PE carriers who developed an infection were analyzed. Results: 6112 patients and 4902 patients were screened during the two periods. 384 and 232 ESBL-PE carriers were identified. Total number of infectious episodes were 146 and 114, respectively. A total of 144 pneumonias, 42 urinary tract infection and 45 digestive infections were studied. An ESBL-PE was involved in 35 (24.3%) episodes of pneumonia, and 44 (37.9%) of extra-pulmonary infections. The most frequent ESBL-PE involved were K. pneumoniae, E. cloacae and E. coli. Similar species and phenotypes were present in colonisation and infection in 29 (82.8%) of pneumonia and in 40 (90.9%) of extra-respiratory infection. Multivariate analysis identified Klebsiella pneumonia or Enterobacter cloacae carriage as risk factor for ESBL-PE involvement in pneumonia and E. coli carriage and detection of ESBL-PE carriage before ICU admission as protective factors. Conclusion: In our study an ESBL-PE involvement is infrequent in pneumonia. A known carriage before ICU admission and E. coli carriage are factors associated with the absence of ESBL-PE un the episode of respiratory infection. A confirmation of our findings could lead to a reduction in the empirical use of carbapenems in this population.

Background: Chronic rhinosinusitis (CRS) is a heterogeneous inflammatory airway disease associated with impaired mucociliary clearance and persistent inflammation. While prior work has focused on inflammatory and molecular pathways, the physicochemical properties of mucus itself remain poorly characterized. This study aimed to define compositional and biophysical features of CRS mucus that may contribute to dysfunction. Methods: A prospective cross-sectional study was conducted in 15 adults undergoing endoscopic sinus surgery (11 CRS, 4 controls). Mucus was collected from the middle meatus. Hydration was measured by lyophilization. Ionic composition was quantified using mass spectrometry. Viscoelasticity was assessed via oscillatory shear rheology. Total protein, total carbohydrate, sialic acid (Sia) and fucose (Fuc) content were quantified using enzymatic and chemical assays. Statistical comparisons were performed using nonparametric tests. Results: CRS mucus exhibited significantly higher Ca2+; and Mg2+; concentrations (approximately two-fold; p<0.05) and increased variability in hydration and ion content compared to controls. Rheology showed greater heterogeneity and a non-significant trend toward increased viscoelasticity in CRS. Total protein and carbohydrate content were not significantly different; however, the carbohydrate-to-protein ratio was significantly reduced in CRS (p=0.04). Sia content and Sia-to-carbohydrate ratio were significantly elevated in CRS (p=0.04 and p=0.002), particularly in CRS with nasal polyps. Fuc content did not differ between groups. Conclusions: CRS mucus demonstrates coordinated alterations in ionic composition and glycosylation, characterized by increased cation content, hypersialylation, and reduced carbohydrate-to-protein ratios. These changes may contribute to altered mucus properties and impaired mucociliary clearance, highlighting mucus composition as a potential therapeutic target in CRS.

BackgroundBlood transfusion is a life-saving intervention; however, transfusion-transmissible infections (TTIs) such as human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), and syphilis remain major public health concerns, particularly in low-and middle-income countries. This study assessed the seroprevalence and temporal trends of TTIs among blood donors in the Volta Region of Ghana and to identify the demographic factors associated with seropositivity MethodsA retrospective cross-sectional study was conducted using secondary data from blood donors at Ho Teaching Hospital and Hohoe Regional Hospital between January 2020 and December 2023. Data from 6,147 eligible donors were extracted and analyzed using STATA version 17. Descriptive statistics summarized prevalence, while chi-square or Fishers exact tests assessed associations. Multivariable logistic regression was used to identify predictors of TTI seropositivity at a 5% significance level. ResultsThe overall prevalence of TTIs was 8.1%, with syphilis (3.6%) being the most prevalent infection, followed by HBV (1.8%), HCV (1.8%), and HIV (1.0%). All infections peaked in 2022 before declining in 2023. Older age ([&ge;]50 years) and year of donation were significant predictors of TTI positivity. In Hohoe, male donors had lower odds of HCV infection compared to females (aOR = 0.13; 95% CI: 0.06-0.28). ConclusionsAlthough TTI prevalence was relatively low, temporal increases and age-related disparities highlight the need for strengthened donor screening, targeted recruitment of voluntary donors, and enhanced surveillance strategies to ensure blood safety.

Aim: Cross-sectional summaries of periodontitis based on clinical attachment loss (CAL) are, by definition, conditioned on surviving teeth. Because the most severely affected teeth are more likely to have been lost, these measures may underestimate cumulative disease burden and show an artificial flattening (attenuation) of severity with age. We hypothesised that measures more sensitive to severe attachment loss would show greater attenuation at older ages than measures defined across a broader range of sites. Materials and Methods: Using nationally representative data from adults aged 30+ years in NHANES 2009-2014, we examined age-specific trajectories across multiple continuous measures of periodontal severity and assessed whether divergence between measures followed the pattern predicted under severity-dependent tooth loss. Results: The proportion of observable sites declined from 93% at ages 30-34 to 68% at 80+ years, establishing the structural basis for the divergence observed across severity measures. All severity measures showed nonlinear attenuation with age, with distortion increasing with severity threshold. Higher-threshold measures exhibited the greatest attenuation, while lower-threshold measures showed more stable trajectories. Conclusions: Cross-sectional summaries of periodontitis reflect disease among surviving teeth rather than cumulative damage across teeth originally at risk. Attenuation at older ages is consistent with depletion of the most severely affected teeth rather than biological slowing. Distortion varies by measure, with higher-threshold and mean-based indices most affected, whereas the CAL 3+ mm threshold provides a more stable basis for age comparisons.

IntroductionPressure sores are a major health problem in people with spinal cord injury resulting in ischaemic tissue lesions caused by prolonged pressure against a bony surface. Conventional therapies are often defective and fundamental researches on the healing process of pressure sores must be enriched in order to understand any novel therapies that may be applied. We focalize on pressure sores fibroblasts as dermal fibroblasts perform a critic role in wound healing by populating the wound site to produce extracellular matrix. After characterizing morphological and the genetic profile of healthy fibroblasts and fibroblasts from pressure ulcers, we conducted an analysis of fibroblast proliferation, migration and myofibroblastic differentiation capacity. Materials and Methodsafter acquisition of dermal explants and fibroblasts culture, we conducted histological analysis, an evaluation of gene expression by RT-qPCR and an assessment of fibroblasts proliferation and migration capacity through IncuCyte. A study of the differentiation of fibroblasts into myofibroblasts through the detection of Alpha-Smooth Muscle Actin (-SMA) expression by immunofluorescence was also conducted. Resultshistological analysis showed histological analysis showed dermal disorganization in pressure sore compared with health skin, differences in morphological aspects and density of fibroblasts. Pressure sore fibroblasts express less genes coding for ECM proteins, metalloproteases, collagen III, Connective tissue growth factor (CTGF) and ACTA2 coding for -SMA. Pathological fibroblasts appear to proliferate less quickly than healthy fibroblasts but no differences in migration capacity were found. After stimulation under TGF-{beta}, pressure sore fibroblasts lose their ability to differentiate into myofibroblasts compared to healthy fibroblasts and this could be in relation with a less expression of ACTA2. ConclusionAll of our results highlight a morphological, genetic and functional difference between healthy and pathological fibroblasts which have a modified phenotype, less effective for skin repair. This suggests that new therapies for chronic wounds must take into account the environment in which they are applied and that pathological cells do not necessarily respond to treatments in the same way as healthy cells. Our results are not statistically significant, although several trends emerge. This is explained by the heterogeneity of the patients medical history and requires repetition of the experiments.

RationaleSepsis is a life-threatening syndrome causing significant morbidity and mortality especially in the aging population. Clonal hematopoiesis of indeterminate potential (CHIP) is an age-related condition of clonal expansion of hematopoietic stem cells harboring somatic mutations associated with increased incidence of chronic illness and all-cause mortality. ObjectiveEvaluate the association of pre-illness CHIP with mortality and morbidity in patients admitted to the ICU with sepsis. MethodsWe performed a retrospective study using a de-identified electronic health record linked with a DNA biorepository. We identified adult patients with sepsis who had DNA collected prior to ICU admission. We tested the association between CHIP status, determined from whole-genome sequencing, and ICU mortality, organ support-free days, and long-term survival adjusting for age, sex, race and Sequential Organ Failure Assessment (SOFA) score on ICU admission. Measurements and Main ResultsPre-illness CHIP was associated with increased sepsis mortality (OR = 1.54, 95% CI 1.13 to 2.07, P = 0.005) and fewer days alive and free of organ support (-1.7 days, 95% CI -3.2 to -0.2, P = 0.028) after adjusting for age, sex, race, and SOFA score. In sepsis survivors, CHIP was also associated with increased long-term mortality after discharge (HR 1.40, 95% CI 1.01 to 1.93, P = 0.041). ConclusionsPre-illness CHIP was independently associated with increased mortality and morbidity in critically-ill adults with sepsis. These findings suggest that CHIP is a risk factor for sepsis severity. Elucidating the mechanism underlying this association could uncover new therapeutic interventions for sepsis.

Abstract / SummarySurvival outcomes for pediatric Burkitt lymphoma (BL) substantially vary depending on geography (50-90%), which also serves as a proxy for the prevalence of Epstein-Barr virus (EBV) within the tumors. Although BL is considered an immunologically "cold" tumor with few tumor-infiltrating lymphocytes (TILs), their functional status has not been fully evaluated, especially for EBV-positive disease. Here, we characterize the exhaustion and activation profiles of T cells in the tumor microenvironment (TME) of EBV-positive BL using orthogonal methods, single-cell gene expression analysis, spectral flow cytometry, and immuno-histochemistry staining (IHC). We found that CD8+ TILs displayed a mosaic of immune inhibitory gene expression encoding, PD1, TIGIT, LAG3 and HAVCR2/TIM3. IHC validated the expression of PD1 and TIGIT on CD8+ TILs, as well as their respective ligands, PDL-1, PVR, and Nectin-2 on malignant B cells. Despite exhaustion-associated signatures, CD8+ TILs retain cytotoxic potential, expressing granules (i.e. Granzyme A, Perforin) and cytokines (i.e. IFN{gamma}) and demonstrate an increased uptake of metabolites such as glucose, arginine, and methionine. In peripheral blood, pediatric BL patients exhibited a significantly higher abundance of PD1+TIGIT+ CD8+ T cells compared to healthy children. Notably, these circulating T cells from BL patients express significantly lower levels of TOX, suggesting they are not irreversibly dysfunctional. Together, our results indicate that CD8+ T cells both in the TME and in circulation of children with BL are not terminally exhausted but remain poised for functional re-invigoration. These findings support the potential integration of immune checkpoint inhibitors into combination chemotherapeutic regimens to improve outcomes for these children. SignificanceEBV-positive BL tumors contain functional, metabolically active CD8+ T cells. Circulating PD1+TIGIT+CD8+ T cells found in BL patients blood are a biomarker for those in the tumor microenvironment.

Vaccine-induced immune responses are the result of an intricate interplay between different cell populations of the innate and adaptive immune system, which is so far only partly understood. In particular, the role of polymorphonuclear neutrophils (PMNs) has long been neglected. Here, we studied the effects of a whole inactivated virus influenza vaccine (WIV) in an in vitro system consisting of freshly isolated human PMNs alone or PMNs combined with autologous peripheral blood mononuclear cells (PBMCs). Isolated PMNs showed minimal responses to the vaccine with respect to apoptosis, gene expression, cytokine production, and reactive oxygen species production. However, in WIV-stimulated PMN/PBMC co-cultures, PMNs particularly enhanced monocyte dynamics, CD14-CD11c+ cell activation, effector T cell differentiation, and B cell antibody production. On the other hand, PMNs decreased T follicular helper cell frequencies. Without vaccine stimulation, PMN presence resulted in enhanced levels of baseline inflammatory cytokines in PMN/PBMC co-cultures. However, with vaccine stimulation, PMNs dampened the vaccine-induced cytokine secretion of PBMCs. These findings reveal PMNs as regulators of vaccine responses whose effects depend on crosstalk with other immune cells, balancing pro-inflammatory and adaptive immune activation. Author summaryPolymorphonuclear neutrophils (PMNs) are essential and predominant cells of the human innate immune system. Growing evidence implicates that PMNs are involved in vaccine-induced immune activation, but their exact role is so far poorly defined. In our study, human PMNs were tested alone to observe their response to whole inactivated virus influenza vaccine (WIV), or combined with autologous peripheral blood mononuclear cells (PBMCs) to investigate how their presence influences vaccine responses of various cell populations within PBMCs. Our results show that WIV had little direct effect on isolated PMNs. However, when PMNs were combined with other immune cells, PMNs acted as crucial regulators: they enhanced the activity of innate immune cells, regulated the responses to the vaccine of T and B cells, and helped control the overall level of inflammation. Our study forms the groundwork for a more comprehensive understanding of human immune cell interactions under vaccine stimulation.

Background: Plasma proteomics may identify host-response signatures in sepsis, but it is unclear whether extracellular vesicle (EV)-enriched plasma provides distinct or redundant information compared with plasma. We compared paired plasma and EV-enriched plasma proteomes in critically ill patients with sepsis and critically ill non-sepsis controls (CINS). Methods: In this prospective observational study, paired plasma and EV-enriched plasma samples were analyzed from 56 critically ill adults, including 40 patients with sepsis and 16 CINS patients. Protein abundance was quantified using liquid chromatography-tandem mass spectrometry. Analyses compared proteomic depth, protein overlap, global concordance between compartments, and differential protein abundance between CINS and sepsis. Exploratory Gene Ontology enrichment was performed as a supplementary analysis. Results: EV-enriched plasma expanded proteomic detection, identifying 2,476 filtered proteins compared with 506 in plasma. Only 386 proteins were detected in both compartments, while 2,090 were unique to EV-enriched plasma and 120 were unique to plasma. Among shared proteins, plasma and EV-enriched plasma showed modest global concordance across critically ill patients (Spearman coeff = 0.322, p = 9.19 x 10^-11), with similar findings in sepsis alone. Differential abundance analysis identified 11 sepsis-associated proteins in plasma and 22 in EV-enriched plasma. Only SAA1, SAA2, and IGFBP6 were significant in both compartments. Exploratory pathway analysis supported acute-phase and inflammatory enrichment in plasma sepsis-associated proteins, while EV-enriched signals were directionally plausible but did not meet prespecified FDR thresholds. Conclusion: Plasma and EV-enriched plasma proteomics capture related but nonredundant sepsis-associated host-response information in critically ill patients.

IntroductionTo investigate the effects of chronic hypoxic exposure simulating heavy-industrial environments on the viability, metabolic activity, stemness preservation, and osteogenic differentiation potential of human dental pulp stem cells (hDPSCs). MethodsCommercially available hDPSCs were cultured under controlled oxygen tensions representing surface atmospheric conditions (21% O2), moderate hypoxia (10% O2), deep hypoxia (5% O2), and severe hypoxia (1% O2). Cells were maintained for 1, 3, and 7 days. Cell viability was evaluated using MTT and Live/Dead assays. Reactive oxygen species (ROS) accumulation, mitochondrial membrane potential, and apoptosis were assessed using fluorescent probes and Annexin V/PI staining. Stemness marker expression (SOX2, OCT4, NANOG) and osteogenic markers (RUNX2, ALP, OCN) were analyzed via RT-qPCR. ResultsModerate hypoxia (10% O2) promoted transient increases in stemness marker expression and preserved metabolic activity. Severe hypoxia (1% O2) significantly reduced cell viability, increased ROS accumulation, disrupted mitochondrial integrity, and elevated apoptotic cell populations after prolonged exposure (p < 0.05). Osteogenic differentiation markers were significantly downregulated under severe hypoxic conditions. ConclusionsIndustrial hypoxic environments critically influence pulpal stem cell physiology and regenerative potential. While moderate oxygen reduction may transiently preserve stemness characteristics, chronic severe hypoxia impairs viability and osteogenic functionality. Chronic low-oxygen occupational environments may alter endogenous dental regenerative mechanisms and influence oral tissue healing responses.

Background: Periodontitis is defined by cumulative, irreversible tissue destruction, yet population-based measurement typically relies on cross-sectional indicators derived from retained teeth. Destruction that occurred earlier in life, particularly disease severe enough to result in tooth loss, is structurally excluded from these measures, potentially leading to systematic underestimation of lifetime periodontal burden. Objective: To develop and evaluate a measurement framework that estimates lifetime periodontal burden from cross-sectional data by explicitly incorporating informative tooth loss under etiological uncertainty. Methods: Data were drawn from 10,324 adults aged [&ge;]30 years participating in the 20090-2016 National Health and Nutrition Examination Survey (NHANES) who completed full-mouth periodontal examination and glycated hemoglobin (HbA1c) testing. Lifetime periodontal burden was estimated by combining observed clinical attachment loss in retained teeth with probabilistic contributions from missing teeth, using three alternative age-stratified attribution schedules derived from epidemiological studies of periodontal extraction. Performance was compared with conventional measures of periodontal severity and extent using distributional analyses, correlations with HbA1c, discrimination of diabetes status, and relative importance analysis. Age-adjusted models were treated as sensitivity analyses. Results: Estimated lifetime periodontal burden exhibited strong, monotonic age gradients across glycemic categories, in contrast to more attenuated patterns observed for severity and extent. Across attribution schedules, lifetime burden showed stronger correlations with HbA1c ({rho} = 0.30-0.32) than conventional measures. In multivariable models including all indices, lifetime burden retained an independent association with HbA1c, whereas severity and extent contributed little unique information. Discriminative performance for diabetes status was consistently higher for lifetime burden than for conventional measures and remained stable across attribution schedules. Conclusions: Lifetime periodontal burden can be estimated from cross-sectional data by explicitly modelling informative tooth loss rather than restricting measurement to retained teeth. Incorporating historical tissue loss under uncertainty yields a more coherent representation of cumulative periodontal destruction than snapshot-based measures and provides a methodological basis for life-course-oriented periodontal epidemiology.

BackgroundShort-chain fatty acids (SCFAs) are produced by the gut microbiota as secondary metabolites during fermentation process of dietary fibers. Although SCFAs are beneficial for immuno-related diseases because they regulate the gene expression and functions of myeloid cells, the effects of SCFAs on the development of DCs remain unclear. MethodsWe analyzed the effect of SCFAs on the expression levels of surface proteins and mRNAs, and histone modification in Flt3L-induced bone marrow-derived DCs. ResultsSCFAs, particularly butyrate, regulated the expression of surface molecules on mouse bone marrow-derived dendritic cells (DCs): increases in MHCII, CD86, CD11b, and LPAM-1 (4{beta}7) levels and the ratio of CD11c+/PDCA-1-/B220- conventional DCs (cDCs) to CD11c+/PDCA-1+/B220+ plasmacytoid DCs (pDCs). Experiments using inhibitors of histone deacetylase (HDAC) and Gi proteins, and GPR109A deficient mice indicated that butyrate regulated DCs by suppression of HDACs and not through a stimulatory effect on G protein-coupled receptors. Butyrate and the HDAC inhibitor, trichostatin A (TSA), increased the cDC/pDC ratio, surface LPAM-1 and Itga4 mRNA, while the mRNA level of Itgb7 was not affected by butyrate and was reduced by TSA. ChIP assays showed that butyrate and TSA increased histone acetylation in the Itga4 and Spi1 genes. Furthermore, the butyrate treatment increased the levels of Spi1 mRNA and PU.1 protein and decreased those of Spib/SpiB in DCs. In knockdown (KD) experiments using siRNAs, the gene expression of Itga4 was decreased by KD of Spi1 or Irf8, and cDC/pDC ratio decreased by Spi1 KD. ConclusionsButyrate controls the gene expression and development of DCs through epigenetic regulation and DC-related transcription factors.

BackgroundiPSCs generated from healthy individuals constitute an important control resource for disease modelling applications but existing biobanks are highly skewed towards populations of European ancestry while well characterized control lines from Indian populations remain limited. Given the extensive genetic diversity of the Indian subcontinent, the availability of ethnically relevant healthy control lines is important for developing accurate disease models and reducing population specific confounding effects. MethodologyWe used peripheral blood mononuclear cells (PBMNCs) of a healthy female donor of Eastern Indian origin for the generation a wild type iPSC line using non-integrating episomal reprogramming vectors. Established colonies were expanded and characterized through morphological assessment, expression of pluripotency and trilineage markers, episomal vector clearance analysis, and chromosomal stability evaluation and mycoplasma contamination analysis. ResultsThe line generated exhibited characteristic pluripotent stem cell morphology and also showed strong expression of pluripotency markers, was free from any contamination and free from the reprogramming vectors confirming an integration free system. The cells maintained a normal diploidy number during characterization. Expression of lineage specific markers associated with ectoderm, mesoderm and endoderm confirmed the developed iPSCs functional capacity to undergo trilineage differentiation. ConclusionWe have developed and validated an iPSC line from an underrepresented Indian population. This well characterized, ethnicity specific iPSC line provides a valuable cell line for establishing a high quality, well characterized control baseline, which is a major missing element in South Asian stem cell repositories and thus will provide a solid foundation for future disease specific modelling and screening.

ObjectiveThe objective of this study was to compare hospital length-of-stay and other clinical outcomes between intact fish skin graft (IFSG; Graftguide, Kerecis, Arlington, VA) and synthetic/biosynthetic dermal substitutes (SSS; Integra Dermal Regeneration Template and NovoSorb Biodegradable Temporizing Matrix) in propensity score-matched burn patients using the American Burn Association Burn Care Quality Platform. MethodsThis retrospective cohort study identified adult patients treated with a single dermal substitute product during hospitalization for acute burn injury. Patients receiving IFSG (n = 93) were matched 1:4 to patients receiving SSS (n = 372) using nearest-neighbor propensity score matching on the logit scale. Matching covariates included total body surface area burned (TBSA), patient age, sex), burn severity classification, inhalation injury, and trauma diagnosis. The primary outcome was hospital length of stay (LOS), analyzed using a gamma generalized linear mixed model (GLMM). Secondary outcomes included the incidences of sepsis, graft loss, venous thromboembolism (VTE), and hospital-acquired pressure injury (HAPI). A prespecified sensitivity analysis was performed using a broader mixed-product cohort. ResultsA total of 93 IFSG-treated patients from 17 burn centers admitted between the years 2019 and 2025 were matched 1:4 to 372 SSS-treated patients from 44 centers. Unadjusted mean LOS was 24.1 days (median 20, IQR 11-32) in the IFSG-treated group and 36.7 days (median 31, IQR 17-52) in the SSS-treated group representing a 12.6-day reduction. GLMM-adjusted estimated marginal mean LOS was 24.2 days (95% CI, 20.0-29.4) for IFSG versus 33.5 days (95% CI, 30.0-37.6) for SSS (ratio 0.723; p = 0.00245), representing a 9.3-day reduction. Sepsis (1.1% vs 4.6%), graft loss (3.2% vs 8.3%), VTE (2.2% vs 2.7%), and HAPI (2.2% vs 3.8%) were all numerically lower in the IFSG-treated arm; although GLMM-adjusted odds ratios were not statistically significant for any individual complication. The mixed-cohort sensitivity analysis (n = 229 IFSG vs 458 SSS across 67 centers) confirmed the primary finding with GLMM-adjusted LOS ratio 0.716 (p = 0.0001). ConclusionsIn this propensity score-matched analysis of the ABA registry, IFSG was associated with a statistically significant and clinically meaningful reduction in hospital length of stay compared with synthetic/biosynthetic dermal substitutes, in requiring dermal substitution and autografting, with all complication rates, sepsis, graft loss, VTE, and HAPI, numerically lower in the IFSG-treated arm. The shorter hospitalization was not achieved at the expense of safety. These findings support IFSG as a viable alternative to synthetic dermal substitutes in burns requiring dermal substitution and autografting. Prospective studies are warranted particularly in larger burns requiring staged reconstruction.