Search Results (407)

Several chronic inflammatory processes are currently being studied in relation to other systems to better understand the regulation mechanisms and identify potential therapeutic targets. A significant body of evidence supports the role of the nervous system in regulating various immunological processes. This study investigates the relationship between pterygia and the sympathetic nervous system, focusing on their interaction in the inflammatory response and fibrogenic process. Sixteen surgical specimens of primary pterygia and four conjunctival tissue samples were examined, and their morphology was analyzed using hematoxylin–eosin and Masson’s trichrome stains. The gene expression of adrenergic receptors, as well as inflammatory and fibrogenic cytokines, was also assessed. Additionally, both adrenergic receptors and tyrosine hydroxylase were found to be localized within the tissues according to immunohistochemistry and immunofluorescence techniques. Increased expression of proinflammatory, fibrogenic, and adrenergic genes was observed in the pterygium compared to the healthy conjunctiva. Adrenergic receptors and tyrosine hydroxylase were localized in the basal region of the epithelium and within blood vessels, closely associated with immune cells. Neuroimmunomodulation plays a key role in the pathogenesis of pterygia by activating the sympathetic nervous system. At the intravascular level, norepinephrine promotes the migration of immune cells, thereby sustaining inflammation. Additionally, sympathetic nerve fibers located at the subepithelial level contribute to epithelial growth and the fibrosis associated with pterygia. Full article

Although cranial growth has been extensively explored, forensic and biological anthropology lack a formal incorporation of how cranial growth processes impact the adult phenotype and downstream biological profile estimations. Objectives: This research uses an ontogenetic framework to identify when interlandmark distances (ILDs) stabilize during growth to reach adult levels of variation and to evaluate patterns of cranial sexual size dimorphism. Methods: Multivariate adaptive regression splines (MARS) were conducted on standardized cranial ILDs for 595 individuals from the Subadult Virtual Anthropology Database (SVAD) and the Forensic Data Bank (FDB) aged between birth and 25 years. Cross-Validated R-squared (CVRSq) values evaluated ILD variation explained by age while knot placements identified meaningful changes in ILD growth trajectories. Results: Results reveal the ages at which males and females reach craniometric maturity across splanchnocranium, neurocranium, basicranium and cross-regional ILDs. Changes in growth patterns observed here largely align with growth milestones of integrated soft tissue and skeletal structures as well as developmental milestones like puberty. Conclusions: Our findings highlight the variability in growth by sex and cranial region and move forensic anthropologists towards recognizing cranial growth as a mosaic, continuous process with overlap between subadults and adults rather than consistently approaching subadult and adult research separately. Full article

Micro-fragmented adipose tissue (MFAT) is a promising autologous therapy for knee osteoarthritis. To avoid repeated liposuction procedures for its clinical application, MFAT obtained from patients with knee osteoarthritis was stored at −80 °C in a tissue bank. This study describes the preparation, cryopreservation, thawing, and washing, as well as comprehensive analysis of cell populations in fresh and MFAT thawed after two years. Immunophenotyping of both fresh and thawed MFAT showed a significant presence of endothelial progenitors and pericytes in the stromal vascular fraction. Viability before (59.75%) and after freezing (55.73%) showed no significant difference. However, the average cell count per gram of MFAT was significantly reduced in thawed samples (3.00 × 10⁵) compared to fresh ones (5.64 × 10⁵), likely due to processing steps. Thawed MFAT samples showed increased CD73 expression on the CD31^highCD34^high subset of EP and SA-ASC, as well as increased expression of CD105 on EP, the CD31^lowCD34^low subset of EP, pericytes, and SA-ASC. Microbiological testing confirmed 100% sterility, and double washing efficiently removed DMSO, confirming sample safety. Histological analysis revealed healthy, uniformly shaped adipocytes with intact membranes. This approach allows accurate estimation of cell yield for intra-articular injection, ensuring delivery of the target cell number into the knee. Quality control analysis confirms that cryopreserved MFAT retains high cellular and structural integrity, supporting its safety and suitability for clinical application. Full article

Background: SERPINE1, also known as plasminogen activator inhibitor (PAI), has been proposed as a potential blood biomarker for the early detection and diagnosis of Alzheimer’s disease (AD). Expanding on previous studies, this research contrasted SERPINE1 levels in CSF and brain tissue of AD patients and those at risk for AD with established AD biomarkers. Methods: Utilizing OLINK and immunoassay methods, CSF SERPINE1 protein levels were quantified across two separate cohorts: PREVENT-AD and ADNI. Microarray and RNAseq were used to measure tissue SERPINE1 mRNA levels in two separate cohorts: the Douglas-Bell Canada Brain Bank and the Mayo Clinic Brain Bank. Results: At the pre-clinical stage, elevated CSF levels of pTau, tTau and synaptic markers, alongside reduced hippocampal volume, correlate with CSF SERPINE1 levels. Elevated cortical SERPINE1 mRNA levels in autopsy-confirmed AD show weak correlation with regional plaques and tangles densities, but strong correlation with Braak staging. Conclusions: CSF SERPINE1 levels can be used as an early biomarker for the detection of pathological changes associated with AD. Higher SERPINE1 levels correlate more strongly with tau pathology than with amyloid formation or deposition. Full article

Donor human milk (DHM) can harbor microbial contaminants that cause serious infections in premature infants. Bacillus cereus is a pathogen frequently found in DHM, capable of forming spores that can resist Holder pasteurization (62.5 °C, 30 min). Since no microbial growth is acceptable in post-pasteurized DHM, microbiological testing of pre-pasteurized DHM provides information about its contamination level to determine if it should be accepted for pasteurization. Culture is the gold standard in microbiological control but it requires 24–48 h to provide results. In this study we developed and validated a non-commercial real-time PCR assay for the detection of Bacillus cereus (BC test) in DHM specimens on a fully automated high-throughput platform, the cobas^® 6800 system. The BC test showed excellent sensitivity and specificity, repeatability and linearity over an 8-log range and a low limit of detection in milk specimens, as well as good agreement with selective culture methods. BC test was then used to systematically control all milk donations (3439) over a 24-month period. Bacillus cereus was detected in 14.2% of DHM, with monthly rates ranging from 6 to 29% and a significantly higher incidence in warmer months. Incorporating this assay into our laboratory workflow improved efficiency and reduced turnaround time. Full article

Background and Objectives: To characterize the epidemiological profile of patients undergoing revision total hip arthroplasty (THA) using bone allografts from a tissue bank, and to identify clinical and surgical factors associated with the selection of graft type in cases of severe periprosthetic bone loss. Materials and Methods: This observational, cross-sectional study involved a retrospective review of medical records from a specialized referral center, including revision THA procedures performed between 2013 and 2019. Data were collected on 36 variables covering demographic details (age, sex), surgical history of both hips, comorbidities, medication use, perioperative complications, hospitalization, surgical technique, and characteristics of the bone grafts used. Patients were grouped based on the type of allograft received—structured or morselized (impacted)—and comparative analyses were performed. Results: A total of 67 revision THA cases were evaluated, with a mean patient age of 63.2 years. Nearly half (47.8%) had no prior hip revision. The average number of previous procedures per patient was 1.73, and the mean interval from primary THA to revision was 178.4 months. Morselized bone allografts were used in 66.7% of cases, and structured allografts in 33.3%. Patients receiving structured grafts had undergone a significantly higher number of prior surgeries (p = 0.01) and had a longer duration since the initial THA (p = 0.04). Conclusions: These findings suggest that younger patients undergoing primary total hip arthroplasty may be at increased risk for complex revision procedures involving structured grafts later in life, underscoring the need for long-term monitoring and tailored surgical planning in this population. Full article

Background/Objectives: Although the nanocomposite of poly(L-lactic acid) with graphene oxide (PLLA-GO) shows promise for tissue engineering, its specific bioactive interactions with diverse cell lineages during early tissue regeneration remain unclear. This study comprehensively investigated the in vitro multifaceted biocompatibility of PLLA-GO using human fibroblasts (FN1 cells), murine mesenchymal stem cells (mBMSCs), and human umbilical vein endothelial cells (HUVECs). Methods: Morphological analyses were performed using optical and scanning electron microscopy, while proliferation dynamics were assessed via CFSE staining. Cell cycle progression was evaluated using flow cytometry, mitochondrial activity was examined through TMRE staining, and inflammatory cytokine profiling was performed via Cytometric Bead Array (CBA). Results: PLLA-GO exhibited primary biocompatibility across all evaluated cell lines, characterized by efficient adhesion and proliferation. However, significant cell-type-dependent modulations were observed. The FN1 cells exhibited proliferative adaptation but induced accelerated scaffold degradation, as evidenced by a substantial increase in cellular debris (5.93% control vs. 34.38% PLLA-GO; p = 0.03). mBMSCs showed a transient initial proliferative response and a significant 21.66% increase in TNF-α production (179.67 pg/mL vs. 147.68 pg/mL in control; p = 0.03). HUVECs demonstrated heightened mitochondrial sensitivity, exhibiting a 32.19% reduction in mitochondrial electrical potential (97.07% control vs. 65.82% PLLA-GO; p ≤ 0.05), alongside reductions in pro-inflammatory cytokines TNF-α (8.73%) and IL-6 (12.47%). Conclusions: The PLLA-GO processing method is crucial for its properties and subsequent cellular interactions. Therefore, rigorous and specific preclinical evaluations—considering both cellular contexts and fabrication—are indispensable to ensure the safety and therapeutic potential of PLLA-GO in tissue engineering and regenerative medicine. Full article

The major objective was to investigate the protective capabilities of endophytic bacterial strains isolated from a number of medicinal plant species towards Aspergillus spp. secured from the internal tissues of fungi-infected peanuts. Among 32 fungal isolates surveyed for mycotoxin production in various culture media (PDA, RBCA, YES, CA), 10 isolates qualitatively producing AFB₁, besides 10 OTA-producers, were assayed by HPLC for quantitative toxin production. Aspergillus spp. isolate Be 13 produced an extraordinary quantity of 1859.18 μg mL⁻¹ AFB₁, against the lowest toxin level of 280.40 μg mL⁻¹ produced by the fungus isolate IS 4. The estimated amounts of OTA were considerably lower and fell in the range 0.88–6.00 μg mL⁻¹; isolate Sa 1 was superior, while isolate Be 7 seemed inferior. Based on ITS gene sequencing, the highly toxigenic Aspergillus spp. isolates Be 13 and Sa 1 matched the description of A. novoparasiticus and A. ochraceus, respectively, ochraceus, respectively, which are present in GenBank with identity exceeding 99%. According to 16S rRNA gene sequencing, these antagonists labeled Ar6, Ma27 and So34 showed the typical characteristics of Pseudomonas aeruginosa, Bacillus subtilis and Bacillus velezensis, respectively, with similarity percentages of 99–100. The plant growth-promoting activity measurements of the identified endophytes indicated the production of 16.96–80.00 μg/100 mL culture medium of IAA. Phosphate-solubilizing capacity varied among endophytes from 2.50 to 21.38 μg/100 mL. The polysaccharide production pool of bacterial strains ranged between 2.74 and 6.57 mg mL⁻¹. P. aeruginosa Ar6 and B. velezensis successfully produced HCN, but B. subtilis failed. The in vitro mycotoxin biodegradation potential of tested bacterial endophytes indicated the superiority of B. velezensis in degrading both mycotoxins (AFB₁-OTA) with average percentage of 88.7; B. subtilis ranked thereafter (85.6%). The 30-day old peanut (cv. Giza 6) seedlings grown in gnotobiotic system severely injured due to infection with AFB₁/OTA-producing fungi, an effect expressed in significant reductions in shoot and root growth traits. Simultaneous treatment with the endophytic antagonists greatly diminished the harmful impact of the pathogens; B. velezensis was the pioneer, not P. aeruginosa Ar6. In conclusion, these findings proved that several endophytic bacterial species have the potential as alternative tools to chemical fungicides for protecting agricultural commodities against mycotoxin-producing fungi. Full article

Background: In 2015, Spanish scientific societies published a consensus document on managing massive hemorrhage (MH). This study aimed to evaluate the knowledge and application of the Massive Hemorrhage Protocol (MHP) among healthcare professionals and to assess whether an educational intervention could improve compliance and patient outcomes. Methods: A two-phase observational study was conducted in four Spanish university hospitals. In phase one, compliance with MHP recommendations was surveyed. Based on the findings, educational sessions were implemented, focusing on the least known or followed recommendations. Compliance was then reassessed. Primary outcome was adherence to MHP; secondary outcomes included morbidity and 24 h and in-hospital mortality. Results: The MHP was activated in 303 MH episodes, mostly of surgical (42.6%) or traumatic (25%) origin. The most followed recommendation before the intervention was protocol activation (94%), which improved to 98.3% post-intervention (p = 0.049). Lesser-followed recommendations such as requesting a hemorrhage lab panel and correcting hypothermia improved after intervention from 39% to 50.4% (p = 0.05) and 31.3% to 43.8% (p = 0.027), respectively. Overall compliance increased from 68% to 73% (p = 0.05). Mortality remained high in both phases, 24 h (25.4%) and in-hospital (42.2%). Patients who required massive transfusion had higher mortality (53.6%) than those who did not (35.9%, p = 0.03). Survivors had higher protocol compliance (p = 0.003 at 24 h; p = 0.049 in-hospital). Conclusions: Educational intervention modestly improved adherence to MHP recommendations. Higher compliance was associated with better survival outcomes, supporting the need for targeted educational strategies to enhance protocol implementation and improve care in MH cases. Full article

Background/Objectives: Understanding the diagnostic landscape is essential prior to developing artificial intelligence (AI)-based diagnostic strategies for automating the diagnosis of duodenal biopsies. This study aims to (1) determine the frequencies of different diagnoses seen in endoscopic duodenal biopsies in a large, tertiary referral centre; (2) identify key words on histopathology request forms that could indicate that a biopsy may contain a serious pathology and should not be diagnosed by an AI system; and (3) investigate the proportion of cases described as showing “intraepithelial lymphocytosis” that might be coeliac disease. Methods: To achieve this, we audited 18 months’ worth of duodenal biopsy reports in our centre. Results: A total of 6245 duodenal biopsies were identified, of which 73.76% were normal and at least 8.84% fell within the spectrum of coeliac disease. Additionally, 6.47% were classified as showing non-specific inflammation, 1.86% were adenomas, 0.45% were carcinomas, 0.06% were neuroendocrine tumours, 0.10% were lymphomas, and 0.03% were cases of flat dysplasia, giving a total of 0.64% of dysplastic or malignant diagnoses. Rarer diagnoses included ulceration, Helicobacter pylori infection, giardiasis, lymphangiectasia, transplant rejection, and lymphoma. Furthermore, 227 biopsies (3.63%) showed isolated intraepithelial lymphocytosis, of which 33 cases (14.5%) gave an overall clinicopathological picture of coeliac disease. Conclusions: We present the first long-term audit of all endoscopic duodenal biopsies received by the histopathology department of a tertiary-care facility. The results indicate that a fully automated diagnostic histopathology reporting system able to identify normal duodenal biopsies and biopsies within the spectrum of coeliac disease-associated enteropathy could decrease pathologists’ endoscopic duodenal biopsy workload by up to 80%. Full article

Secondary osteonecrosis (ON) is a common complication in paediatric cancer survivors. Combining multipotent mesenchymal stromal cells (MSCs) with core decompression surgery halts disease progression and stimulates bone regeneration. However, the success of advanced therapy medicinal products (ATMPs) requires versatile “off-the-shelf” tissue engineering products (TEPs). This study evaluated the safety and efficacy of TEPs loaded with allogeneic MSCs from Wharton’s jelly (WJ-MSCs) in a large-animal model of bone regeneration to support a paediatric investigational plan for ON patients. WJ-MSC-laden fibrin-based hydrogels combined with a synthetic bone substitute (PRO-DENSE^TM) were tested in 16 juvenile sheep (8 males and 8 females) distributed in four experimental groups. Each animal received four cylindrical bone defects in the femoral and tibial epiphyses and was assessed at 6 and 12 weeks. Safety was confirmed, and bone regeneration was observed across all groups. A combination of WJ-MSCs with PRO-DENSE^TM led to improved histological scores, osteogenesis, and construct integration. Trabecular bone volume also increased more in cellular groups over time. However, effects were inconsistent across groups, reflecting the variability seen in clinical trials and highlighting the significant impact of factors such as immunogenetic compatibility, MSC batch potency, and interaction with the recipient’s microenvironment on the therapeutic effectiveness and successful clinical translation of allogeneic ATMPs. Full article

Considering the taxonomic uncertainties of Neotropical deer species, as well as the threat status of many of them, new studies and strategies for their maintenance are urgently needed. Obtaining live cells is of great importance for the conservation of wild species in order to allow cytogenetic and molecular studies to be carried out and for the construction of genomic resource banks. In order to increase the genetic diversity stored in these banks, the possibility of collecting skin fragments from dead animals (e.g., run over, hunted, deaths related to disease or natural causes) becomes a valuable source and a last alternative for obtaining material from these individuals. However, the interval between the death of the animal and the collection of tissue can directly interfere with the quality of the sample obtained and it is therefore essential to identify the maximum time during which viable cells are still found. Thus, this study sought to establish a protocol for the collection, storage, cryopreservation, and cultivation of skin obtained postmortem from individuals of the species Subulo gouazoubira (gray brocket deer) and Mazama rufa (red brocket deer). The collection of tissue fragments at different postmortem intervals (0 h, 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 7 h, 8 h, 9 h, 10 h, and 11 h) was evaluated. The tissues were analyzed for fibroblast cell viability immediately after collection. Their ability to undergo cryopreservation was evaluated based on techniques that can be directly applied to samples obtained in the field and their subsequent thawing and success of cell cultures was performed in the laboratory. Regarding the genetic integrity of the cells, the number of metaphases was observed by the mitotic index. The cell viability presented by the samples always remained above 60%. It was possible to establish cell cultures even with the tissues obtained 11 h after the death of the individuals; however, they required twice as many days to reach bottle confluence compared to the cultures performed with the tissues obtained 0 h after the death of the individuals. The results suggest that the best rates of cell viability, time to reach confluence, and number of metaphases per cell (mitotic index) are found in skin fragments collected up to 5 h after the death of individuals when their carcasses are kept at room temperature. Full article

Background: Platelet-rich plasma (PRP) is increasingly utilized for managing knee osteoarthritis (KOA), yet its clinical value remains debated due to the variability in preparation protocols and outcome measures. Methods: This narrative review synthesizes current evidence from 40 high-quality studies published between 2013 and March 2025, including randomized controlled trials, systematic reviews, and meta-analyses. The biological mechanisms, clinical effectiveness, safety, and implementation challenges of PRP therapy in KOA are examined. Results: PRP injections—particularly leukocyte-poor PRP—demonstrate superior pain relief and functional improvement compared to hyaluronic acid and corticosteroids, especially in patients with mild to moderate KOA (Kellgren–Lawrence grades I–III). However, heterogeneity in PRP formulations (platelet/leukocyte content and activation protocols), injection regimens, and follow-up durations limits direct comparability across studies. Evidence from high-quality placebo-controlled trials shows inconsistent long-term benefits, with some failing to demonstrate superiority over saline beyond 6–12 months. The GRADE assessment rates the overall certainty of evidence as moderate. PRP appears safe, with few adverse events reported, but remains costly and variably reimbursed. Guidelines from major societies remain cautious or inconclusive. Conclusions: PRP is a promising, safe, and well-tolerated option for early to moderate KOA. However, the standardization of preparation protocols, patient selection criteria, and outcome reporting is essential to improve comparability and guide clinical practice. Full article

The role of gender in osteoarthritis (OA) has been reported. However, knowledge on whether gender-specific regulatory SNPs are determining factors in OA is limited. We aimed to identify susceptible gender-specific SNPs of transcription factor binding sites in OA. We used a modified NF-κB binding motif from an RNA sequencing data-inferred OA-associated upstream regulator to define genome-wide potential NF-κB binding sites, which were aligned to the Taiwan BioBank SNP database to identify susceptible SNPs. A case-control study was conducted to verify SNPs with OA determined by a logistic model. The functional assessment was validated using the Genotype-Tissue Expression Portal database. We collected 533 OA patients and 614 healthy controls. Two of nine novel OA-associated SNPs were identified to be significant. For males, the variant of rs73164856 in the aldose reductase gene enhancer was identified to be a protective factor of severe OA patients [odds ratio (OR): 0.17, 95% confidence interval (CI): 0.04–0.73]. For females, the variant of the rs545654 in the neuronal NOS (nNOS) gene was identified to be a detrimental factor of severe OA patients (OR: 2.07, 95% CI: 1.15–3.73). The gene expression analysis demonstrated a lower expression of the AKR1B15 gene (p = 0.00019) upon the rs73164856 T allele; meanwhile, it showed a higher expression of the nNOS gene (p = 1.2 × 10⁻¹⁷) upon the rs545654 T allele. This study identifies susceptible gender-specific SNPs of NF-κB binding sites in severe OA and validates the RMCGA, which sheds light on genetic determinants by gender in advanced OA. Full article

Background: Osteoarthritis (OA) is a degenerative joint disease characterized by progressive cartilage breakdown, synovial inflammation, and pain, which leads to significant disability. IAHA is widely used because of its viscoelastic properties, which restore synovial fluid homeostasis and reduce symptoms. However, emerging evidence suggests that IAHA exerts additional biological effects including chondroprotection, inflammatory modulation, oxidative stress reduction, and pain modulation, which may influence disease progression. Objective: This narrative review examines the biological mechanisms underlying IAHA’s role in OA management. The review explored IAHA’s effects on synovial fluid viscoelasticity, inflammatory cytokine modulation, cartilage preservation, oxidative stress regulation, and pain pathways, emphasizing the influence of molecular weight variations on therapeutic efficacy. Additionally, this review evaluates IAHA’s integration into multimodal treatment strategies, its potential disease-modifying effects, and future directions for personalized treatment approaches. Methods: A comprehensive literature review was conducted using PubMed, Cochrane Library, EMBASE, Scopus, and Web of Science for studies published between January 2000 and March 2024. The search focused on IAHA’s molecular, cellular, and biochemical effects in OA and clinical findings assessing its impact on joint function, pain relief, and disease progression. Results: IAHA improves synovial fluid lubrication, reduces proinflammatory cytokines (IL-1β, TNF-α), inhibits matrix metalloproteinases (MMPs), scavenges reactive oxygen species (ROS), and modulates nociceptive pathways. High-molecular-weight IAHA demonstrates superior efficacy in advanced OA, while low-molecular-weight formulations may be better suited for early-stage disease. Although IAHA’s symptom relief is comparable to corticosteroids and NSAIDs, its favorable safety profile and emerging disease-modifying potential support its long-term use in OA management. Conclusions: IAHA represents a multifaceted therapeutic approach bridging symptomatic relief and regenerative strategies. While long-term efficacy, optimal administration protocols, and patient-specific responses remain subjects of ongoing research, refining treatment selection criteria, dosing regimens, and combination strategies may enhance clinical outcomes. Future studies should explore biomarker-driven approaches, standardize treatment protocols, and assess IAHA’s synergy with regenerative medicine to optimize its role in OA management. Full article