bluuebunny/crossref_metadata_2025 huggingface dataset:
Ophthalmic In Situ Nanocomposite Gel for Delivery of a Hydrophobic Antioxidant
Authors: Slavkova Marta , Voycheva Christina , Popova Teodora , Tzankov Borislav , Tzankova Diana , Spassova Ivanka , Kovacheva Daniela , Stefanova Denitsa , Tzankova Virginia , Yoncheva Krassimira
Published: February 2025

The topical administration of in situ hydrogels for ocular pathologies is a promising application strategy for providing high effectiveness and patient compliance. Curcumin, a natural polyphenol, possesses all the prerequisites for successful therapy of ophthalmic diseases, but unfortunately its physicochemical properties hurdle the practical use. Applying a composite in situ thermoresponsive hydrogel formulation embedded with polymer nanoparticles is a potent strategy to overcome all the identified drawbacks. In the present work we prepared uniform spherical nanoparticles (296.4 ± 3.1 nm) efficiently loaded with curcumin (EE% 82.5 ± 2.3%) based on the biocompatible and biodegradable poly-(lactic-co-glycolic acid). They were thoroughly physicochemically characterized in terms of FTIR, SEM, TGA, and DLS, in vitro release following Fickian diffusion (45.62 ± 2.37%), and stability over 6 months. Their lack of cytotoxicity was demonstrated in vitro on HaCaT cell lines, and the potential for antioxidant protection was also outlined, starting from concentrations as low as 0.1 µM and reaching 41% protection at 5 µM. An in situ thermoresponsive hydrogel (17% w/v poloxamer 407 and 0.1% Carbopol) with suitable properties for ophthalmic application was optimized with respect to gelation temperature (31.40 ± 0.36 °C), gelling time (8.99 ± 0.28 s) upon tears dilution, and gel erosion (90.75 ± 4.06%). Upon curcumin-loaded nanoparticle embedding, the in situ hydrogels demonstrated appropriate pseudoplastic behavior and viscosity at 35 °C (2129 ± 24 Pa∙s), 6-fold increase in the permeation, and prolonged release over 6 h.
Investigation of a Thermoresponsive In Situ Hydrogel Loaded with Nanotriphala: Implications for Antioxidant, Anti-Inflammatory, and Antimicrobial Therapy in Nasal Disorders
Authors: Phongpradist Rungsinee , Chittasupho Chuda , Singh Sudarshan , Ontong Julalak Chorachoo , Tadtong Sarin , Akachaipaibul Puriputt , Punvittayagul Charatda , Thongkorn Kriangkrai , Dejkriengkraikul Pornngarm , Jiaranaikulwanitch Jutamas , Chansakaow Sunee , Hongwiset Darunee
Published: February 2025

Oxidative stress plays a crucial role in chronic nasal disorders, contributing to inflammation, tissue damage, and impaired mucosal function, highlighting the need for targeted therapies. Recent advancements in nasal drug delivery systems have expanded their applications for treating respiratory and inflammatory conditions. Among these, hydrogel-based systems offer prolonged release of active pharmaceutical ingredients (APIs), enhancing therapeutic efficacy and reducing dosing frequency. This study initially evaluates the antioxidant, anti-inflammatory, antimicrobial, and cytotoxic properties of Nanotriphala, followed by its incorporation into a thermoresponsive in situ hydrogel system, which was subsequently developed and characterized as a novel formulation. Nanotriphala exhibited >90% cell viability and significantly reduced nitric oxide (NO) levels by 40.55 µg/mL at 250 µg/mL. The hydrogel was characterized by key parameters, including viscosity, gelling time, pH, gelling temperature, texture analysis, and ex vivo spreadability. Stability was assessed under various conditions, and mutagenicity and antimutagenicity were evaluated using the Ames test. Results showed that the hydrogel gelled at 34 °C, exhibited good spreadability (10.25 ± 0.28 cm), a viscosity of 227 ± 22 cP, and maintained a pH of 5.75 ± 0.01, with optimal hardness and adhesiveness suitable for nasal application. It demonstrated antimicrobial activity against E. coli, P. aeruginosa, S. aureus, and S. epidermidis at minimal bactericidal concentrations (MBCs) of 32, 2, 4, and 8 µg/mL, respectively, with low mutagenicity (mutagenic index < 2) and strong antimutagenic activity (>60%). The gallic acid content was 0.5796 ± 0.0218 µg/100 mL. Stability studies confirmed optimal storage at 4 °C. These findings suggest that in situ hydrogel loaded with Nanotriphala is a promising nasal drug delivery system for managing oxidative stress and related inflammatory conditions.
Evaluation of Bacterial Cellulose/Alginate-Based Hydrogel and Frog Skin Dressings in Equine Skin Wound Healing
Authors: Campebell Rita C. , Oliveira Andressa B. , Fagundes Jéssyca L. A. , Fortes Beatriz N. A. , Veado Henrique C. , Macedo Isabel L. , Dallago Bruno S. L. , Barud Hernane S. , Adorno José , Salvador Pablo A. V. , Santos Paulo S. , Castro Márcio B.
Published: February 2025

This study evaluates the wound-healing process in horses following the application of two treatment modalities: bacterial cellulose hydrogel with alginate (BCAW) and frog skin (FSW) dressings on experimentally induced skin wounds. Throughout the experiment, no clinical abnormalities were noted in the horses, although initial wound assessments indicated edema and sensitivity. Local hemorrhage was observed in some cases on Day 0, with granulation tissue formation evident by Day 14. Epithelialization began around Day 14 but did not reach complete healing in any group by Day 28. The analysis showed no significant differences in skin wound area or wound contraction rates among the treatment groups compared to control wounds (CWs) over the evaluation periods. Histopathological evaluations also indicated no significant differences in inflammatory responses or healing markers, such as fibroblast proliferation and neovascularization in skin wounds across groups. Despite expectations based on prior research in other species, the treatments with BCAW and FSW did not demonstrate substantial pro-healing effects in horses with induced skin wounds. These findings underscore the complexity of equine wound healing and suggest further investigation is needed to optimize treatment strategies in this species and enhance the translational potential for human clinical applications.
An Actively Homing Insertion Element in a Phage Methylase Contains a Hidden HNH Endonuclease
Authors: Arsenault Danielle , Gosselin Sophia P. , Gogarten Johann Peter
Published: February 2025

Background/Objectives: The ShiLan domain was previously identified as an insertion sequence in a phage DNA methylase gene that exhibited similar evolutionary patterns to that of an active intein or self-splicing intron but could not be identified as either. It produces no internal stop codons when read in frame with its host methylase gene, leading to the thought that it may not be an intron and rather be an abnormal type of intein. However, the sequence has no detectable self-splicing domains, which are essential for intein persistence, as preventing an intein from successfully splicing is often detrimental to proper host protein function. Methods: The analysis of alternate open reading frames for the full nucleotide sequence of this insertion element revealed the insertion to be an out-of-frame histidine-asparagine-histidine (HNH) endonuclease. A GTG start codon is located 18 bp into the insertion, and a TAA stop codon within the last four bases of the insertion (TAAC). When this frame is read, an HNH endonuclease is revealed. In-depth computational analysis could not retrieve support for this element being any known type of self-splicing element, neither intein nor intron. When read in-frame with the methylase gene, this insertion is predicted to take on a looping structure that may be able to avoid interference with the DNA methylase activity. We performed searches for sequences similar in nature to the inserted out-of-frame HNH and found several in other phages and prokaryotes. We present our survey of these out-of-frame endonuclease insertion elements as well as some speculation on how these endonucleases are getting translated to facilitate their homing activity. Conclusions: These findings expand our understanding of the possible arrangements for and prevalence of unorthodox mobile genetic elements and overlapping open reading frames in phages.
Transcriptomic Analysis Reveals Patterns of Expression of Stage-Specific Genes in Early Apis cerana Embryos
Authors: Su Runlang , Chen Yuhui , Zhu Rui , Ding Guiling , Dong Kun , Feng Mao , Huang Jiaxing
Published: February 2025

Background/Objectives: Apis cerana development is described as comprising four stages: embryo, larva, pupa, and adult. There are significant differences between workers and drones in terms of physiological functions and social roles, and the formation of the organ primordia occurs during the embryonic stage. Therefore, the objective of this study is to investigate the differential expression of and alternative splicing of genes in worker and drone embryos and to explain their unique developmental patterns. Methods: Long-read sequencing (PacBio Iso-Seq) and short-read sequencing (Illumina RNA-Seq) were used to investigate worker and drone embryo gene expression differences in A. cerana across five developmental points (12, 24, 36, 48, and 60 h). Results: The study identified 59,254 common isoforms, with 5744 and 5106 isoforms specific to worker and drone embryos, respectively. Additionally, a new transcript of the csd gene was identified. The number of differentially expressed genes (3391) and differential splicing events (470 genes) peaked at the 24-h embryonic stage. Differential splicing events of csd, dsx, and Y-y were observed in the worker and drone embryos. Conclusions: The gene expression results indicated that the 24-h embryonic point is a critical period for the expression of genes related to developmental and behavioral differences between workers and drones. The findings provide a theoretical basis for future research on the developmental differences between workers and drones.
Earthquake Damage Susceptibility Analysis in Barapani Shear Zone Using InSAR, Geological, and Geophysical Data
Authors: Sharma Gopal , Singh M. Somorjit , Nayak Karan , Dutta Pritom Pran , Sarma K. K. , Aggarwal S. P.
Published: February 2025

The identification of areas that are susceptible to damage due to earthquakes is of utmost importance in tectonically active regions like Northeast India. This may provide valuable inputs for seismic hazard analysis; however, it poses significant challenges. The present study emphasized the integration of Interferometric Synthetic Aperture Radar (InSAR) deformation rates with conventional geological and geophysical data to investigate earthquake damage susceptibility in the Barapani Shear Zone (BSZ) region of Northeast India. We used MintPy v1.5.1 (Miami INsar Timeseries software in PYthon) on the OpenSARLab platform to derive time series deformation using the Small Baseline Subset (SBAS) technique. We integrated geology, geomorphology, gravity, magnetic field, lineament density, slope, and historical earthquake records with InSAR deformation rates to derive earthquake damage susceptibility using the weighted overlay analysis technique. InSAR time series analysis revealed distinct patterns of ground deformation across the Barapani Shear Zone, with higher rates in the northern part and lower rates in the southern part. The deformation values ranged from 6 mm/yr to about 18 mm/yr in BSZ. Earthquake damage susceptibility mapping identified areas that are prone to damage in the event of earthquakes. The analysis indicated that about 46.4%, 51.2%, and 2.4% of the area were low, medium, and high-susceptibility zones for earthquake damage zone. The InSAR velocity rates were validated with Global Positioning System (GPS) velocity in the region, which indicated a good correlation (R2 = 0.921; ANOVA p-value = 0.515). Additionally, a field survey in the region suggested evidence of intense deformation in the highly susceptible earthquake damage zone. This integrated approach enhances our scientific understanding of regional tectonic dynamics, mitigating earthquake risks and enhancing community resilience.
Tectonic Setting and Spatiotemporal Earthquake Distribution in Northern Nubia and Iberia
Authors: Mantovani Enzo , Viti Marcello , Tamburelli Caterina , Babbucci Daniele , Baglione Massimo , D’Intinosante Vittorio
Published: February 2025

The spatiotemporal distribution of major earthquakes in the study area (1600–2024) is analyzed to tentatively recognize the possible connections with the short-term (from decades to centuries) evolution of the ongoing tectonic processes. This study suggests that during the period considered, seismic activity has been predominantly related to the shortening processes accommodating the convergence of northwestern Nubia with the Iberian and Moroccan plates that mainly involve the westward escape of the Alboran wedge and the NNE-ward escape of the Iberian block. This deformation pattern is inferred from the seismic activity in the North Atlantic domain, the Rif and Betics belts, the western Iberian fault system (onshore and offshore), the Transmoroccan fault system and the Pyrenean thrust front. Seismic activity in the Tell is mainly driven by the Nubia–Eurasia convergence, even though it can be also influenced by the major westward displacements of the Anatolian–Aegean–Adriatic–Pelagian system. This hypothesis could explain the marked increase in seismic activity that occurred in the Tell in the last decades, when that zone may have been affected by the perturbation triggered by the large post-1939 westward displacement of Anatolia. The pieces of evidence and the arguments reported in this study might provide insights into the possible spatial distribution of major earthquakes in the next decades.
Mindfulness Meditation for Sleep Disturbances Among Individuals with Cognitive Impairment: A Scoping Review
Authors: Chan Sunny H. W. , Cheston Richard , Steward-Anderson Charlotte , Yu Chong-Ho , Dodd Emily , Coulthard Elizabeth
Published: February 2025

Background: This scoping review investigates the effectiveness of mindfulness meditation in alleviating sleep disturbances among individuals with mild cognitive impairment (MCI) and Alzheimer’s disease (AD). With the rising prevalence of dementia and its profound impact on cognitive function and quality of life, this review aims to synthesize existing research and identify gaps in the literature. Methods: We systematically searched six electronic databases (CINAHL, Embase, Medline, PsycINFO, PubMed, and Scopus) from 2004 to 2024, yielding 462 potentially relevant articles. Screening was conducted using ASReview, an AI ranking tool, which facilitated the selection of studies. Ultimately, seven studies that met our stringent eligibility criteria were included in the review. We adhered to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) guidelines for reporting. Results: Our findings indicate that mindfulness meditation significantly improves sleep quality, reduces insomnia severity, and enhances overall well-being in this at-risk population. Notably, interventions that combine structured, face-to-face sessions with at-home practice emerged as the most effective. Conclusions: Despite these positive outcomes, methodological limitations, including small sample sizes and reliance on self-reported measures, underscore the need for more rigorous long-term studies. This review highlights the potential of mindfulness meditation as a low-cost, scalable intervention to improve sleep and cognitive health in older adults, paving the way for future research and clinical applications.
Prurigo Pigmentosa Associated with a Ketogenic Diet in a Romanian Patient: A Case Report
Authors: Iliescu Carmen Andrada , Beiu Cristina , Tebeică Tiberiu , Popa Liliana Gabriela
Published: February 2025

Background/Objectives: Prurigo pigmentosa (PP) is a rare inflammatory dermatosis characterized by recurrent pruritic eruptions resolving with reticulated hyperpigmentation. Initially described in young Asian women, PP is increasingly reported worldwide, often linked to ketosis-inducing factors such as low-carbohydrate diets. This report documents the first case of PP in a Romanian patient and highlights the importance of recognizing the condition in diverse populations. Methods: We describe a 16-year-old girl with a six-month history of pruritic eruptions on the neck, trunk, shoulders, and thighs. A detailed clinical assessment was conducted, including dietary history, physical examination, laboratory evaluation of urinary ketones, and skin biopsy for histopathological analysis. Results: Clinical examination revealed erythematous papules and plaques with peripheral pustules and post-inflammatory hyperpigmentation. Substantial weight loss and elevated urinary ketone levels suggested dietary-induced ketosis from a strict low-carbohydrate diet. Histopathological findings confirmed the diagnosis of PP. The patient’s skin lesions resolved following carbohydrate reintroduction and doxycycline therapy. Conclusions: This case highlights the association between ketosis and PP, emphasizing the importance of thorough dietary history-taking and histopathological confirmation for accurate diagnosis. As ketogenic diets become more popular globally, heightened awareness of PP is essential to ensure timely diagnosis and effective management.
Tetanus Vaccine Knowledge, Beliefs, and Attitudes Among Syrian Pregnant Women in Türkiye: A Qualitative Study
Authors: Altaş Zeynep Meva , Abdulhaq Bayan , Sezerol Mehmet Akif , Karabey Selma
Published: February 2025

Background: In this qualitative study, we aimed to evaluate the knowledge, beliefs, and attitudes of Syrian pregnant women living in Türkiye toward tetanus vaccination, vaccine hesitations, and the perceived benefits and harms of the tetanus vaccine. Methods: In face-to-face, in-depth interviews, an open-ended, semi-structured interview guide was used by an interviewer and translator who spoke Arabic, which is the mother tongue of the participants. Codes, themes, and subthemes were created. Results: In the study, face-to-face in-depth interviews were conducted with a total of 16 pregnant women. The median age of the pregnant women was 26.5 years (18.0–41.0). The median time since they arrived in Türkiye as migrants was 8.5 years (3.0–10.0). The themes were “Beliefs about vaccines”, “Information about vaccines”, “Knowledge, beliefs and attitudes about tetanus vaccine”, “Reasons for vaccination desire”, “Reasons for not being vaccinated”, and “Problems experienced while receiving health services”. Pregnant women mostly mentioned that vaccines have benefits. Most of the women stated that tetanus vaccine is needed to protect from diseases. There were no women thinking that vaccines are harmful. Some of them only mentioned the minor side effects observed after vaccination. Participants mentioned that they get information about vaccines from relatives such as family and friends, health professionals, television, and social media. However, some women talked about the fact that they had no knowledge about vaccines. A lack of knowledge and not visiting the health institution were the reasons for not being vaccinated. Conclusions: Although participants mostly believed that vaccination is necessary and has benefits, some women had no idea about the exact purpose of vaccines. This lack of knowledge may cause vaccine hesitancy and refusal.
The Predisposition of Men Who Have Sex with Men to Use Post-Exposure Prophylaxis for HIV in a Capital City in Northeast Brazil
Authors: de Castro Pereira Chaves André Felipe , da Cunha Soares Yndiara Kássia , de Melo Júnior Eugênio Barbosa , de Lima Brito Magalhães Rosilane , Lima Shirley Verônica Melo Almeida , de Tarso Moura Borges Paulo , de Araújo Telma Maria Evangelista
Published: February 2025

The aim of this study was to analyze the predisposition and factors associated with the use of Post-Exposure Prophylaxis (PEP) for HIV in men who have sex with men (MSM). This was a cross-sectional study conducted in the city of Teresina, Piauí, Brazil, between January and July 2024. The study sample consisted of 320 MSM. A questionnaire consisting of 37 previously validated questions and a risk perception scale for HIV with eight questions were used. To explain which factors would be associated with predisposition to the use of PEP, a logistic regression analysis was applied with an odds ratio. The criterion for including variables in the logistic model was an association at the 20% level (p ˂ 0.20) in the bivariate analysis. Statistical significance in the final model was set at 5%. Although the vast majority of MSM reported a willingness to use PEP (94.4%), their knowledge about prophylaxis and their HIV risk perceptions were largely unsatisfactory. It was found that living alone reduces the chances of predisposition to PEP use by 75% (AOR = 0.25; p = 0.01), and using a condom during oral sex reduces the chances of predisposition to PEP use by 91% (AOR = 0.09; p < 0.001). In light of this, the importance of greater investments in health education actions that reinforce the mechanisms of HIV transmission, as well as the use of methods for its prevention, is highlighted. In addition, targeted interventions are needed to improve knowledge about PEP and HIV risk perception.
Determinants of Safe Pesticide Handling and Application Among Rural Farmers
Authors: Oshingbade Olamide Stephanie , Moda Haruna Musa , Akinsete Shade John , Adejumo Mumuni , Hassan Norr
Published: February 2025

The study investigated the determinants of safe pesticide handling and application among farmers in rural communities of Oyo State, ssouthwestern Nigeria. A cross-sectional design utilizing 2-stage cluster sampling techniques was used to select Ido and Ibarapa central Local Government Areas and to interview 383 farmers via a structured questionnaire. Data were analyzed using descriptive statistics and logistic regression at p = 0.05. Results showed that 41.8% of the farmers had been working with pesticides on farms for at least 5 years, 33.0% attended training on pesticide application, 73.5% had good safety and health knowledge, and 72.3% had safe pesticide handling and application practices. About half (50.2%) stated that they wear coveralls, gloves, and masks to protect their body, face, and hands when applying pesticides, 9.8% use empty pesticide containers for other purposes in the house/farm, while 11.5% blow the nozzle with their mouth to unclog it if it becomes blocked. The three major health symptoms reported by the participants were skin irritation (65.0%), itchy eyes (51.3%), and excessive sweating (32.5%). Having attended training on pesticide application and use enhanced (OR = 2.821; C.I = 1.513–5.261) practicing safe pesticide handling and application. Farmers with good knowledge (OR = 5.494; C.I = 3.385–8.919) were more likely to practice safe pesticide handling and application than those with poor knowledge about pesticide use. It is essential to develop and deliver mandatory comprehensive training programs for farmers on impacts of pesticides on health and environment, along with sustainable safe handling, application, and disposal of pesticides using proper waste management techniques and recognizing early signs and seeking medical assistance. The urgent need to strengthen policy to regulate pesticide use and limit farmers’ access to banned products is also key.
The Relationship Between Methadone and Buprenorphine Enrollment and Overdose Prevention and Treatment Behaviors Among a Community Sample of People Who Use Opioids in Baltimore, Maryland
Authors: Latkin Carl A. , Dayton Lauren , Davey-Rothwell Melissa , Jones Abenaa
Published: February 2025

Background: Methadone and buprenorphine can reduce overdose-related mortality. Behavioral approaches can also reduce fatal overdoses. The current study examined the relationship between methadone and buprenorphine and overdose history and overdose prevention and treatment behaviors. Methods: Between December 2022 and August 2024, 647 individuals who used opioids in the prior month enrolled in a community recruited study on overdose. Participants were administered a face-to-face survey. Key behaviors assessed included overdose recency, testing drugs for potency, ingesting drugs slowly, using fentanyl test strips, using drugs alone, and carrying naloxone. Chi-square and logistic regression models examined the relationships between methadone and buprenorphine and overdose-related outcomes. Results: In total, 32.9% of participants were currently taking methadone and 15.5% buprenorphine. Most (69.2%) reported ever overdosing, and among those, 33.7% had overdosed within the prior 6 months. There were no significant associations between methadone or buprenorphine status and overdose prevention and care behaviors. In the multivariable logistic regression model, methadone use was associated with a lower odds ratio (aOR = 0.49, 95% CI = 0.30–0.79of a recent overdose compared to buprenorphine. Daily or almost daily crack use was associated with greater odds of a recent overdose (aOR = 2.21, 95% CI = 1.44–3.39. Discussion: Findings suggest the importance of promoting overdose prevention and care behaviors to people in drug treatment and training them to promote overdose prevention and care behaviors among their drug-using network members and other community members.
Plasma-Activated Water Improve Wound Healing in Diabetic Rats by Influencing the Inflammatory and Remodelling Phase
Authors: Rajić Jovana , Grdović Nevena , Marković Anđelija , Škoro Nikola , Dinić Svetlana , Uskoković Aleksandra , Arambašić Jovanović Jelena , Đorđević Marija , Sarić Ana , Vidaković Melita , Puač Nevena , Mihailović Mirjana
Published: February 2025

Diabetic foot ulcers have an enormous impact on patients’ quality of life and represent a major economic burden. The cause is delayed and incomplete wound healing due to hyperglycemia, reduced blood flow, infections, oxidative stress and chronic inflammation. Plasma-activated water (PAW) is emerging as a new therapeutic approach in wound treatment, as it has many of the advantages of cold atmospheric plasma but is easier to apply, thus allowing for widespread use. The aim of this study was to investigate the potential of PAW to improve wound healing in diabetic rats, with a focus on uncovering the underlying mechanisms. Two full-thickness wounds in control and diabetic animals were treated with PAW, and healing was monitored for 15 days at five time points. PAW improved wound healing in diabetic rats and mainly affected the inflammatory phase of wound healing. Application of PAW decreased the number of inflammatory cells, myeloperoxidase (MPO) and N-acetyl-b-D-glycosaminidase (NAG) activity, as well as the mRNA expression of proinflammatory genes in diabetic rats. Ten days after injury, PAW treatment increased collagen deposition in the diabetic animals by almost 10% without affecting collagen mRNA expression, and this is in correlation with a decrease in the Mmp-9/Timp-1 ratio. In conclusion, PAW treatment affects wound healing by reducing the inflammatory response and influencing extracellular matrix turnover, suggesting that it has great potential to accelerate the healing of diabetic wounds.
Assessment of Protein Immunoexpression Associated with Tumor Proliferation and Invasion in Histological Subtypes of Unicystic and Conventional Ameloblastoma
Authors: da Silveira Gabriela Cristina Avertano Rocha , Costa Rebeca Vieira , Lemos Flavia Letícia Magalhães , de Moraes Antonia Taiane Lopes , Kataoka Maria Sueli da Silva , Freitas Vanessa Morais , de Menezes Silvio Augusto Fernandes , Vasconcelos Ana Carolina Uchoa , Etges Adriana , Santos Fabricio Passador , de Araújo Vera Cavalcanti , Alves Júnior Sérgio de Melo , Jaeger Ruy Gastaldoni , Pinheiro João de Jesus Viana
Published: February 2025

The aim of this study was to verify whether the expression of proteins related to the formation of invadopodia, MT1-MMP, cortactin, Tks-4 and Tks-5 is associated with the degree of tumor invasiveness of different types of unicystic ameloblastomas. An immunohistochemical study was performed on 29 unicystic ameloblastoma (UA) samples, 9 conventional ameloblastoma (CAM) samples and 9 dental follicle (DF) samples. The potential for tumor invasiveness was assessed based on the immunoexpression of the following invadopodia-forming proteins: MT1-MMP, cortactin, Tks-4 and Tks5. Mural unicystic ameloblastoma (MUA) showed higher MT1-MMP, cortactin, Tks-4, and Tks-5 immunoexpression than luminal and intra-luminal types. Conventional ameloblastoma exhibited lower MT1-MMP, cortactin, and Tks-5 expression compared to MUA. MUA’s cystic capsule neoplastic cells had significantly higher MT1-MMP, cortactin, Tks-4, and Tks-5 expression than lumen cells. Dental follicles showed minimal expression. Neoplastic cells in the cystic capsule of mural unicystic ameloblastomas showed higher invadopodia-related protein expression than lumen and luminal/intraluminal cells, suggesting that proximity to the bone region influences the aggressive behavior of mural unicystic ameloblastomas more compared to other subtypes.
Association Between Active DNA Demethylation and Liver Fibrosis in Individuals with Metabolic-Associated Steatotic Liver Disease (MASLD)
Authors: Barchetta Ilaria , Zampieri Michele , Cimini Flavia Agata , Dule Sara , Sentinelli Federica , Passarella Giulia , Oldani Alessandro , Karpach Katsiaryna , Bacalini Maria Giulia , Baroni Marco Giorgio , Reale Anna , Cavallo Maria Gisella
Published: February 2025

Metabolic-associated steatotic liver disease (MASLD) represents the most common chronic hepatopathy worldwide and an independent risk factor for cardiovascular disease and mortality, particularly when liver fibrosis occurs. Epigenetic alterations, such as DNA methylation, may influence MASLD susceptibility and progression; yet mechanisms underlying this process are limited. This study aimed to investigate whether active DNA demethylation in peripheral blood mononuclear cells (PBMCs) from individuals with MASLD, alongside the methylation and mRNA levels of inflammation- and fibrosis-related candidate genes, is associated with liver fibrosis. For this study, global demethylation intermediates (5-hydroxymethylcytosine [5hmC], 5-formylcytosine [5fC]) were quantified in PBMCs from 89 individuals with/without MASLD using ELISA. Site-specific DNA methylation of SOCS3, SREBF1, and TXNIP was analyzed by mass spectrometry-based bisulfite sequencing; mRNA expression was assessed via RT-PCR. Individuals with MASLD and moderate-to-high fibrosis risk (estimated by the fibrosis non-alcoholic steatohepatitis (NASH) index, FNI) progressively exhibited greater global 5hmC and 5fC levels. Higher FNI was associated with reduced methylation of the SOCS3 gene and increased mRNA expression of the SOCS3, TXNIP, IL-6, and MCP-1 genes. In conclusion, elevated fibrosis risk in MASLD is associated with active global DNA demethylation, as well as differential methylation and expression patterns of genes, which are key regulators of inflammation and fibrosis. These epigenetic alterations in PBMCs may mirror DNA methylation changes in the liver, which may potentially contribute to liver fibrogenesis and represent novel biomarkers for MASLD progression toward fibrosis.
mRNA Degradation as a Therapeutic Solution for Mucopolysaccharidosis Type IIIC: Use of Antisense Oligonucleotides to Promote Downregulation of Heparan Sulfate Synthesis
Authors: Santos Juliana Inês , Gonçalves Mariana , Almeida Matilde Barbosa , Rocha Hugo , Duarte Ana Joana , Matos Liliana , Moreira Luciana Vaz , Encarnação Marisa , Gaspar Paulo , Prata Maria João , Coutinho Maria Francisca , Alves Sandra
Published: February 2025

Mucopolysaccharidosis type IIIC is a neurodegenerative lysosomal storage disorder (LSD) characterized by the accumulation of undegraded heparan sulfate (HS) due to the lack of an enzyme responsible for its degradation: acetyl-CoA:α-glucosaminide N-acetyltransferase (HGSNAT). Classical treatments are ineffective. Here, we attempt a new approach in genetic medicine, genetic substrate reduction therapy (gSRT), to counteract this neurological disorder. Briefly, we used synthetic oligonucleotides, particularly gapmer antisense oligonucleotides (ASOs), to target the synthesis of the accumulated compounds at the molecular level, downregulating a specific gene involved in the first step of HS biosynthesis, XYLT1. Our goal was to reduce HS production and, consequently, its accumulation. Initially, five gapmer ASOs were designed and their potential to decrease XYLT1 mRNA levels were tested in patient-derived fibroblasts. Subsequent analyses focused on the two best performing molecules alone. The results showed a high inhibition of the XYLT1 gene mRNA (around 90%), a decrease in xylosyltransferase I (XT-I) protein levels and a reduction in HS storage 6 and 10 days after transfection (up to 21% and 32%, respectively). Overall, our results are highly promising and may represent the initial step towards the development of a potential therapeutic option not only for MPS IIIC, but virtually for every other MPS III form. Ultimately, the same principle may also apply to other neuropathic MPS.
Regulatory T Cell- and Natural Killer Cell-Mediated Inflammation, Cerebral Vasospasm, and Delayed Cerebral Ischemia in Aneurysmal Subarachnoid Hemorrhage—A Systematic Review and Meta-Analysis Approach
Authors: Pfnür Andreas , Mayer Benjamin , Dörfer Lena , Tumani Hayrettin , Spitzer Daniel , Huber-Lang Markus , Kapapa Thomas
Published: February 2025

Aneurysmal subarachnoid hemorrhage (SAH) involves a significant influx of blood into the cerebrospinal fluid, representing a severe form of stroke. Despite advancements in aneurysm closure and neuro-intensive care, outcomes remain impaired due to cerebral vasospasm and delayed cerebral ischemia (DCI). Previous pharmacological therapies have not successfully reduced DCI while improving overall outcomes. As a result, significant efforts are underway to better understand the cellular and molecular mechanisms involved. This review focuses on the activation and effects of immune cells after SAH and their interactions with neurotoxic and vasoactive substances as well as inflammatory mediators. Particular attention is given to clinical studies highlighting the roles of natural killer (NK) cells and regulatory T cells (Treg) cells. Alongside microglia, astrocytes, and oligodendrocytes, NK cells and Treg cells are key contributors to the inflammatory cascade following SAH. Their involvement in modulating the neuro-inflammatory response, vasospasm, and DCI underscores their potential as therapeutic targets and prognostic markers in the post-SAH recovery process. We conducted a systematic review on T cell- and natural killer cell-mediated inflammation and their roles in cerebral vasospasm and delayed cerebral ischemia. We conducted a meta-analysis to evaluate outcomes and mortality in studies focused on NK cell- and T cell-mediated mechanisms.
Subgenomic RNA Detection in SARS-CoV-2 Assessing Replication and Inactivation Through Serial Passages, RT-qPCR, and Electron Microscopy
Authors: França Talita da Silva , da Silva Juliana Fernandes Amorim , da Silva Gabriella Christine Neves , dos Santos Barbara Oliveira , Silva Stephanie Almeida , Linhares José Henrique Resende , da Silva Marcos Alexandre Nunes , Barreto-Vieira Debora Ferreira , de Paula Vanessa Salete , de Morais Liliane Monteiro , Santos Renata Tourinho , Trindade Gisela Freitas
Published: February 2025

Subgenomic RNAs (sgRNAs) are potential markers of active SARS-CoV-2 replication, serving as templates for the synthesis of structural and accessory proteins in infectious viral particles. This study aimed to use RT-qPCR to quantify sgRNA and negative RNA intermediates, assessing viral replication in virus samples inactivated by β-propiolactone (βPL). Inactivated viruses subjected to five blind serial passages (BSs) were amplified by RT-qPCR using primers to target the envelope (ENV) and nucleoproteins (N1 and N2) of genomic genes, subgenomic envelope RNA (sgENV), and intermediate envelope RNA (ENV-). All positive controls showed consistent viral titers across passages (10 log10 copies/mL in N1/N2 and 11 log10 copies/mL in ENV) during BSs. Inactivated viral samples for ENV and ENV- targets ranged from 11.34 log10 copies/mL in BS1 to 11.20 log10 copies/mL in BS5. The sgENV was no longer detected in the inactivated SARS-CoV-2 samples after the second passage, suggesting successful inactivation. Replication kinetics showed consistent profiles for N1/N2, ENV, and ENV- targets in the first three post-infection hours (pih) and maintained approximately 5 log10 copies/mL at 1 pih, 2 pih, and 3 pih. A sharp exponential increase in the viral titer was observed from 24 pih onwards, peaking at 11.64 log10 copies/mL at 48 pih. Transmission electron microscopy confirmed viral particles only in cells infected with active SARS-CoV-2. These results support the use of sgRNA as a reliable marker for SARS-CoV-2 replication, especially in distinguishing between active replication and non-viable particles and in the development of diagnostic and therapeutic strategies.
Impact of High-Temperature Stress on Maize Seed Setting: Cellular and Molecular Insights of Thermotolerance
Authors: Fan Zhaoyi , Song Haoqi , Qi Mengyuan , Wang Mengqing , Bai Yunfeng , Sun Yuhui , Yu Haidong
Published: February 2025

Global warming poses a significant threat to crop production and food security, with maize (Zay mays L.) particularly vulnerable to high-temperature stress (HTS). This review explores the detrimental impacts of elevated temperatures on maize development across various growth stages, analyzed within the source–sink framework, with a particular focus on seed setting and yield reduction. It provides a broad analysis of maize cellular and molecular responses to HTS, highlighting the key roles of plant hormone abscisic acid (ABA) signaling, calcium signaling, chloroplast, and the DNA damage repair (DDR) system in maize. HTS disrupts ABA signaling pathways, impairing stomatal regulation and reducing water-use efficiency, while calcium signaling orchestrates stress responses by activating heat shock proteins and other protective mechanisms. Chloroplasts, as central to photosynthesis, are particularly sensitive to HTS, often exhibiting photosystem II damage and chlorophyll degradation. Recent studies also highlight the significance of the DDR system, with genes like ZmRAD51C playing crucial roles in maintaining genomic stability during reproductive organ development. DNA damage under HTS conditions emerges as a key factor contributing to reduced seed set, although the precise molecular mechanisms remain to be fully elucidated. Furthermore, the review examines cutting-edge genetic improvement strategies, aimed at developing thermotolerant maize cultivars. These recent research advances underscore the need for further investigation into the molecular basis of thermotolerance and open the door for future advancements in breeding thermotolerant crops.
Rational Identification of Ritonavir as IL-20 Receptor A Ligand Endowed with Antiproliferative Properties in Breast Cancer Cells
Authors: Maggisano Valentina , Gargano Adriana , Maiuolo Jessica , Ortuso Francesco , De Amicis Francesca , Alcaro Stefano , Bulotta Stefania
Published: February 2025

Targeting the tumor microenvironment (TME) is an attractive strategy for developing new drugs with anticancer activity against triple-negative breast cancer (TNBC). Interleukins (ILs) are key players in the TME cytokine network promoting cancer progression. Recent studies have highlighted the involvement of IL-20 receptor subunit alpha (IL-20RA) signalling in several cancers, including BC, in which IL-20RA is highly expressed, correlating with poor prognosis and influencing tumoral characteristics such as proliferation, cell death, invasiveness, and TME activity. Therefore, elucidating the role of the IL-20RA signalling pathway could form the basis for developing new therapeutic strategies. This study aimed to identify selective bioactive ligands able to affect IL-20RA activity. Virtual screening of over 310,000 compounds from both the DrugBank and ZINC15 databases identified four potential hit compounds tested for their anticancer activity against TNBC in vitro cell lines. Notably, Ritonavir, a well-known Human Immunodeficiency Virus Type 1 (HIV-1) protease inhibitor, significantly inhibited cell proliferation (about 40% at 50 µM, p < 0.001). IL-20 preincubation counteracted Ritonavir’s cytostatic effect while IL-20RA knockdown restored proliferation in Ritonavir-treated TNBC cells. In conclusion, these findings demonstrated that Ritonavir reduced TNBC cell proliferation through IL-20RA activity modulation, suggesting its potential repurposing as a therapeutic agent for TNBC management.
The Potential of Single-Transcription Factor Gene Expression by RT-qPCR for Subtyping Small Cell Lung Cancer
Authors: Iñañez Albert , del Rey-Vergara Raúl , Quimis Fabricio , Rocha Pedro , Galindo Miguel , Menéndez Sílvia , Masfarré Laura , Sánchez Ignacio , Carpes Marina , Martínez Carlos , Pérez-Buira Sandra , Rojo Federico , Rovira Ana , Arriola Edurne
Published: February 2025

Complex RNA-seq signatures involving the transcription factors ASCL1, NEUROD1, and POU2F3 classify Small Cell Lung Cancer (SCLC) into four subtypes: SCLC-A, SCLC-N, SCLC-P, and SCLC-I (triple negative or inflamed). Preliminary studies suggest that identifying these subtypes can guide targeted therapies and potentially improve outcomes. This study aims to evaluate whether the expression levels of these three key transcription factors can effectively classify SCLC subtypes, comparable to the use of individual antibodies in immunohistochemical (IHC) analysis of formalin-fixed, paraffin-embedded (FFPE) tumor samples. We analyzed preclinical models of increasing complexity, including eleven human and five mouse SCLC cell lines, six patient-derived xenografts (PDXs), and two circulating tumor cell (CTC)-derived xenografts (CDXs) generated in our laboratory. RT-qPCR conditions were established to detect the expression levels of ASCL1, NEUROD1, and POU2F3. Additionally, protein-level analysis was performed using Western blot for cell lines and IHC for FFPE samples of PDX and CDX tumors, following our experience with patient tumor samples from the CANTABRICO trial (NCT04712903). We found that the analyzed SCLC cell line models predominantly expressed ASCL1, NEUROD1, and POU2F3, or showed no expression, as identified by RT-qPCR, consistently matching the previously assigned subtypes for each cell line. The classification of PDX and CDX models demonstrated consistency between RT-qPCR and IHC analyses of the transcription factors. Our results show that single-gene analysis by RT-qPCR from FFPE-extracted RNA simplifies SCLC subtype classification. This approach provides a cost-effective alternative to IHC staining or expensive multi-gene RNA sequencing panels, making SCLC subtyping more accessible for both preclinical research and clinical applications.
Exploiting Silica-Binding and Silica-Forming Proteins as Versatile Tools for One-Step Enzyme Immobilization on Siliceous Materials
Authors: Lim Gyun Taek , Jo Byung Hoon
Published: February 2025

Enzyme immobilization has emerged as an essential technique in industrial applications of enzymes. Silica (SiO2) serves as a prominent support material for enzyme immobilization. Recent advancements have led to the development of various silica-binding proteins (SBPs) and silica-forming proteins (SFPs) that are invaluable tools in immobilizing enzymes on siliceous materials in a fast and simple manner. SBPs facilitate the immobilization of enzymes with controlled orientation on silica surfaces, while SFPs enable the biomimetic synthesis and encapsulation of enzymes within silica particles. In this review, we explore recent advances in the use of SBPs and SFPs in enzyme applications. We provide a comprehensive overview of their mechanisms and sequence characteristics relevant to enzyme immobilization. Additionally, we summarize the recombinant production and immobilization procedures for enzymes with SBPs or SFPs. We then categorize the available SBPs and SFPs into naturally occurring and artificially engineered types, presenting recent examples that demonstrate their utilization in enzyme immobilization. Our review highlights the strengths and limitations of various SBPs and SFPs and sheds light on future directions for the development of tailor-made biocatalytic silica.
Impact of Rearing Substrates on Black Soldier Fly Growth and Fertility: A Semi-Industrial Scale Study to Optimize Egg Collection
Authors: Zhang Qi-Hui , Puniamoorthy Nalini
Published: February 2025

Juvenile environments can influence adult phenotypes in holometabolous insects. This study examines the effects of larval rearing substrates on the reproductive outcomes of Black Soldier Flies (BSFs) at a semi-industrial scale. Larvae were reared on five substrates. Larval growth, adult size, and reproductive performance were monitored, with a specific focus on egg production and hatching rates across 17 continuous egg collection days. Egg production was positively correlated with female body weight, while male body weight appeared to influence the egg production peak. Extended larval feeding stages and shorter non-feeding periods were associated with an earlier onset of egg-laying, suggesting life-history traits as predictors for reproductive timing. Diets high in carbohydrates but low in protein and fat positively influenced larval growth, egg production, and female body weight, while diets rich in both protein and carbohydrates negatively affected all measured parameters. Notably, higher fecundity did not necessarily correlate with fertility (i.e., more viable larvae for bioconversion), highlighting the need to optimize diets for both egg quantity and hatchability to maximize productivity. These findings provide valuable insights for optimizing larval nutrition and egg collection practices, enhancing reproductive efficiency and overall productivity in industrial BSF farming.
A Genome-Wide Analysis of Nuclear Mitochondrial DNA Sequences (NUMTs) in Chrysomelidae Species (Coleoptera)
Authors: He Yeyan , Ge Siqin , Liang Hongbin
Published: February 2025

Nuclear mitochondrial DNA sequences (NUMTs) are mitochondrial DNA fragments in the nuclear genome, and their unclear distribution in Chrysomelidae species hinders the selection of accurate molecular markers for species identification and phylogenetic analysis. Our study presents a genome-wide survey of NUMTs in 32 Chrysomelidae species. Filtering strategies based on sequence length and open reading frame (ORF) features were employed to identify mitochondrial protein-coding genes (PCGs) minimally affected by NUMTs. Phylogenetic relationships were inferred from both mitochondrial PCG datasets and a COX1 dataset containing NUMTs. Our results show that NUMTs are chromosomally specific, species-specific, and widely distributed. ATP8, COX1, ND1, and ND4 are identified as relatively reliable molecular markers. Phylogenetic analysis is influenced by NUMTs and other factors such as sequence type and saturation. A total of 66 independent COX1 gene nuclear integration events were estimated across 32 species, mostly from distinct mitochondrial lineages. These findings suggest that NUMTs reflect key evolutionary processes such as gene flow and mitochondrial lineage diversification. Their prevalence emphasizes the need for refined molecular markers in species identification and phylogenetic analysis, while also highlighting the importance of NUMTs in understanding mitochondrial DNA integration and their contribution to species’ evolutionary history.