Nanocarrier-based drug delivery systems (DDS) offer promising strategies to enhance therapeutic efficacy, stability, and targeted delivery of bioactive compounds while reducing side effects. In this study, a multidisciplinary approach was employed to develop and characterize bioactive liposomes incorporating resveratrol (RV), a polyphenol with known therapeutic potential but limited bioavailability, and Thymus vulgaris essential oil (TEO), recognized for its antimicrobial and antioxidant properties. To obtain stable nanocarriers, liposomes were formulated using soy lecithin and prepared by thin layer evaporation technique followed by sonication process. To assess the preservation of TEO's chemical profile during formulation, Headspace Solid Phase Microextraction coupled with Gas Chromatography-Mass Spectrometry (HS-SPME/GC-MS) was performed. Quantitative analysis of thymol and carvacrol, the two main active components, was performed by HPLC-UV experiments. Physicochemical characterization of DDS was carried out employing Dynamic Light Scattering (DLS), UV-Vis spectrophotometer and Transmission electron microscopy. Antimicrobial activity was assessed by broth microdilution method, while biofilm inhibition ability was evaluated using crystal violet staining. The obtained nanocarriers exhibited appropriate physicochemical characteristics, including optimal particle size and stability. They can efficiently load both oil and RV. These features enhanced the antibacterial activity of LT-RV compared to free RV, and improved its inhibitory effect, at sub-MIC concentrations, against the sessile form of Listeria monocytogenes strains. This study highlights the importance of integrating chemical, physical, and biological evaluations to develop effective nanocarrier-based formulations with the aim to identify a suitable strategy to combat infections associated with resistant, biofilm-producing pathogens, and offer a versatile and effective delivery platform.

Therapeutic strategies to limit doxorubicin (DOX)-induced cardiomyopathy are still limited due to incomplete characterization of the underlying molecular mechanisms. The exogenous activation of autophagy was reported to exert cardioprotective effects in preclinical models of cardiovascular diseases. We tested whether restoration of autophagy by different pharmacological approaches can reduce DOX-induced cardiomyopathy. A validated preclinical murine model of DOX-induced cardiotoxicity (final cumulative dose of 15 mg/kg) was used to test the cardiac effects of both natural (trehalose and spermidine) and synthetic (Tat-Beclin 1 D11) activators of autophagy. Cardiac function was evaluated by echocardiographic analyses. We performed histological analyses (Masson trichrome staining, TUNEL assay) to investigate fibrosis and apoptosis. Biomolecular analyses, confocal and transmission electron microscopy were used to assess levels of autophagy, autophagic flux, mitophagy and mitochondrial alterations. We also evaluated the effects of autophagy activators in a validated syngeneic model with subcutaneous injection of breast cancer cells treated with DOX. We found that DOX-induced cardiotoxicity is associated with impaired autophagic flux (accumulation of LC3-II and p62). Reactivation of autophagic flux by trehalose or spermidine rescues cardiac function and cardiomyocyte survival in mice treated with DOX, along with an overall amelioration of mitochondrial health and mitophagy. Selective reactivation of autophagy by Tat-Beclin 1 D11 also recapitulates the protective effects exerted by trehalose and spermidine. Autophagy activators preserve cardiac function without affecting the antineoplastic effects of DOX in mice with cancer. Boosting autophagic flux is a suitable therapeutic approach to prevent cardiotoxicity induced by anthracyclines.

The possibility of modifying the surface chemistry of materials and synthetizing inorganic particles in natural aqueous extracts of plants (avoiding calcination), opens the doors to undoubtedly interesting scenarios for innovative functionalization strategies that are increasingly eco-sustainable and rich in interesting chemical–physical and biochemical properties. Among the aerial plants, Satureja montana exhibits interesting antibacterial, antifungal, antimicrobial, and antioxidant activities due to the rich volatile and non-volatile compounds (characterized by gas chromatography–mass spectrometry), contained in the aqueous extracts. For the first time, the latter was applied for the green synthesis of TiO2 and CaCO3 particles, characterized by X-ray diffraction, Raman, infrared spectroscopies, and scanning electron microscopy, coupled with microanalysis. Screening through antimicrobial assays under indoor passive sedimentation conditions showed opposite trends for two kinds of particles. TiO2 anatase spherical particles (400 < φ < 600 nm) increase microbial growth, proportionally to increasing particle concentration. Instead, S. montana-functionalized CaCO3 prismatic microparticles (1 µm × 1 µm × 1 µm) exhibit strong and dose-dependent antimicrobial activities, achieving near-complete inhibition at 50 mg/mL.

Abstract Immune dysregulation plays a vital role in colorectal cancer initiation and progression. Long noncoding RNAs (lncRNA) exhibit multiple functions including regulation of gene expression. Here, we identified an immune-related lncRNA, MIR17HG, whose expression was gradually upregulated in adjacent, adenoma, and colorectal cancer tissue. MIR17HG promoted tumorigenesis and metastasis in colorectal cancer cells both in vitro and in vivo. Mechanistically, MIR17HG increased the expression of NF-κB/RELA by competitively sponging the microRNA miR-375. In addition, RELA transcriptionally activated MIR17HG in a positive feedback loop by directly binding to its promoter region. Moreover, miR-17-5p, one of the transcribed miRNAs from MIR17HG, reduced the expression of the tumor suppressor B-cell linker (BLNK), resulting in increased migration and invasion of colorectal cancer cells. MIR17HG also upregulated PD-L1, indicating its potential role in immunotherapy. Overall, these findings demonstrate that MIR17HG plays an oncogenic role in colorectal cancer and may serve as a promising therapeutic target. Significance: These findings provide mechanistic insight into the role of the lncRNA MIR17HG and its miRNA members in regulating colorectal cancer carcinogenesis and progression.

The increasing global demand for sustainable food production, coupled with the micronutrient limitations of aquaponic systems, calls for innovative fertilization strategies that enhance crop yield and nutritional quality while preserving environmental integrity. This study evaluates the effectiveness of zinc-functionalized calcium carbonate microparticles (Zn-Alg-CaCO₃ MPs) as a smart foliar fertilizer in aquaponics. Tomato plants (Solanum lycopersicum L. cv. Principe Borghese) were grown in a recirculating aquaponic system and treated with three concentrations of Zn-MPs (10, 50, and 250 ppm) via foliar spray, using a control group treated with water. Each treatment was applied to 15 plants under controlled greenhouse conditions. Yield parameters, antioxidant content, and nutritional quality were assessed. The 250 ppm treatment significantly increased yield per plant (+ 74.9%), number of fruits (+ 44.4%), and average fruit weight (+ 22.1%) compared to control. Antioxidant activity, phenolic, flavonoid, and lycopene content were also enhanced, particularly at the highest dose. Importantly, no increase in zinc was detected in recirculating water, highlighting the environmental safety of foliar application. These findings underscore the potential of Zn-Alg-CaCO₃ MPs as a sustainable solution for precision fertilization in aquaponic agriculture. Further molecular and environmental studies are recommended to optimize and scale this technology.

Background: Cholesteatoma is a destructive middle ear pathology requiring precise surgical removal to prevent recurrence and preserve auditory function. The chemically assisted dissection (CADISS) system (AuXin Surgery, Ottignies-Louvain-la-Neuve, Belgium), based on Mesna (5%), was introduced to enhance tissue separation and minimize residual disease. Objective: This study aimed to compare the cleaning efficiency of CADISS-assisted dissection versus the conventional manual dissection of cholesteatoma from incus bone surfaces using quantitative ultrastructural analysis. Methods: This retrospective study evaluated 67 human incus samples collected during cholesteatoma surgery-35 treated with manual dissection and 32 with CADISS. Samples were imaged using variable pressure scanning electron microscopy (VP-SEM) in hydrated conditions. Clean area/total area ratios were calculated and analyzed statistically using non-parametric tests. Postoperative MRI follow-up at 1 month was conducted to assess residual disease. Results: CADISS-assisted samples demonstrated significantly higher clean area/total area ratios (mean: 0.2095 vs. 0.0478, p < 0.0001). Qualitative imaging showed fewer residuals > 1 mm in the CADISS group (9% vs. 77%). MRI follow-up revealed a lower recurrence rate in the CADISS group (3.1%) compared to manual dissection (11.4%). Conclusions: CADISS-assisted dissection provides superior cholesteatoma debris removal compared to manual methods, as evidenced by VP-SEM imaging and clinical follow-up. This technique may improve surgical outcomes and reduce recurrence risk in middle ear surgery.

To evaluate the cleaning efficacy of different irrigation activation techniques in removing smear layers from root canals. Ninety lower premolars with straight root canals were assigned to six experimental groups (n = 15 each): control group, conventional irrigation, passive ultrasonic activation (PUI), distilled water laser-activated irrigation (LAI), PulpSucker irrigation, and iVac irrigation. Each canal was shaped to size 30/04 and irrigated with 5

Colorectal cancer (CRC) is a major public health concern and identifying prognostic molecular biomarkers can help stratify patients based on risk profiles, thus enabling personalized medicine. Epitranscriptomic modifications play a relevant role in controlling gene expression, N6-methyladenosine (m6A) regulators play crucial roles in cancer progression, but their clinical significance in CRC cancer has thus far not been elucidated. Thus, we aimed to examine by immunohistochemical techniques and RT-qPCR, protein levels and RNAs expression of m6A writers (METTL3, WTAP) and eraser (FTO) in a cohort of 10 patients affected by CRC. The patients were followed for 5 years and values of METTL3, WTAP and FTO RNAs in alive vs dead patients were compared. Proteins expression and RNAs expression had a different trend, METTL3, WTAP and FTO proteins' expression showed an increasing trend from non-cancerous adjacent (N) tissue vs carcinoma (CA) tissue G1 stage, and then a decreasing trend from G1 to G2 and G3 stages. The most marked increase was observed in WTAP that, from a 40% of protein expression positivity in N tissue raised to the 81% of positivity in G1 stage K tissue. RNAs expression of METTL3, WTAP and FTO genes in N tissue vs G1 stage CA tissue was significantly different, the analysis and comparison of RNAs values in patient alive after 5 years (0.58±0.04) vs patients dead after 5 years (1.69±0.29) showed that only WTAP values resulted significantly high in dead patients. The fact that WTAP protein expression levels lower while WTAP RNA expression remains high, lets us hypothesize a sort of inhibition of protein expression, but further studies are needed to clarify the mechanism. Although the results suggest a relationship between biological meaning and prognostic utility of WTAP, this prognostic utility must be confirmed by further studies on a larger sample.

Background:In adults, the cerebellopontine angle (CPA) pilocytic astrocytoma (PA) is very rare. This tumor has radiological features similar to those of a vestibular schwannoma in the few cases reported in the literature. Methods:In this study, we conducted a systematic review in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses protocol and scrutinized all original studies pertaining to pontocerebellar angle PA in adult patients. We conducted an analysis of the clinical, radiological, and molecular components of all eligible articles. We have also reported a case involving a 67-year-old male individual in whom the PA exhibited radiological characteristics similar to an epidermoid cyst. Results:After the screening phase, we found four cases of PA of the pontocerebellar angle. Three cases were identified that resembled vestibular schwannoma; however, in our case, the tumor resembled an epidermoid cyst. These uncommon tumors exhibit distinctive histological patterns and molecular characteristics (adenosine triphosphate dependent helicase (ATP- dependent helicase)+, Isocitrate dehydrogenase 1-), rendering them a potential differential diagnosis for glioblastoma (GBM). Conclusion:The CPA PA has rarely been found in adult patients and should be considered in the differential diagnosis of vestibular schwannoma and epidermoid cysts. In these rare cases, the histological characteristics of PA are significant for the differential diagnosis of GBM.

The gut microbiota is an integral component of the colorectal cancer (CRC) microenvironment and is intimately associated with CRC initiation, progression, and therapeutic outcomes. We reviewed recent advancements in utilizing nanotechnology for modulating gut microbiota, discussing strategies and the mechanisms underlying their design. For future nanomedicine design, we propose a 5I principle for individualized nanomedicine in CRC management.

This study investigated the marginal adaptation of three recently introduced bioceramic root repair materials, EdgeBioCeramic RetroFill, Endocem MTA, and One-Fil PT, using VP-SEM analysis. Extracted single-rooted lower incisors were used to simulate retrograde fillings. The results showed no statistically significant differences in the marginal gap between the materials and the dentin walls. All three materials exhibited good dimensional stability, with gap sizes comparable to previously published research on similar materials. The mean GAP was 3.91 ± 2.56 for EdgeBioCeramic RetroFill, 4.32 ± 2.69 for Endocem MTA, and 4.50 ± 2.54 for One-Fil PT. This study employed VP-SEM, a valuable tool for analyzing bioceramic materials without altering their properties. The findings suggest the possibility of daily clinical use of these bioceramics by endodontists and general practitioners that could find applications in retrograde fillings and perforation repairs. However, further in vivo studies are needed to confirm long-term stability and assess the influence of sample preparation methods.

Since the discovery of graphene, there has been a wide range of the literature dealing with its versatile structure and easy binding of biomolecules as well as its large loading capacity. In the emerging field of immunotherapy, graphene and its derivatives have potential uses as drug delivery platforms directly into tumour sites or as adjuvants in cancer vaccines, as they are internalized by monocytes which in turn may activate adaptive anti-tumoral immune responses. In this study, we expose cells of the innate immune system and a human acute monocytic leukemia cell line (THP-1) to low doses of small-sized GO nanosheets functionalized with bovine serum albumin (BSA) and fluorescein isothiocyanate (FITC), to study their acute response after internalization. We show by flow cytometry, uptake in cells of GO-BSA-FITC reaches 80% and cell viability and ROS production are both unaffected by exposure to nanoparticles. On the contrary, GO-BSA nanosheets seem to have an inhibitory effect on ROS production, probably due to their antioxidant properties. We also provided results on chemotaxis of macrophages derived from peripheral blood monocytes treated with GO-BSA. In conclusion, we showed the size of nanosheets, the concentration used and the degree of functionalization were important factors for biocompatibility of GO in immune cells. Its low cytotoxicity and high adaptability to the cells of the innate immune system make it a good candidate for deployment in immunotherapy, in particular for delivering protein antigens to monocytes which activate adaptive immunity.

Background: Spinal cord lymphomas represent a minority of extranodal lymphomas and often pose diagnostic challenges by imitating primary spinal tumors or inflammatory/infective lesions. This paper presents a unique case of primary cauda equina lymphoma (PCEL) and conducts a comprehensive review to delineate the clinical and radiological characteristics of this rare entity. Case Report: A 74-year-old male presented with progressive paresthesia, motor weakness, and symptoms indicative of cauda equina syndrome. Neurological examination revealed paraparesis and sphincter dysfunction. Imaging studies initially suggested an intradural meningioma. However, surgical intervention revealed a diffuse large B-cell lymphoma infiltrating the cauda equina. Findings: A systematic review of the pertinent literature identified 18 primary cauda equina lymphoma cases. These cases exhibited diverse clinical presentations, treatments, and outcomes. The mean age at diagnosis was 61.25 years for women and 50 years for men, with an average follow-up of 16.2 months. Notably, 35% of patients were alive at 18 months, highlighting the challenging prognosis associated with PCEL. Discussion: Primary spinal cord lymphomas, especially within the cauda equina, remain rare and diagnostically complex due to their nonspecific clinical manifestations. The review highlights the need to consider spinal cord lymphoma in patients with neurological symptoms, even without a history of systemic lymphoma. Diagnostic Approaches: Magnetic resonance imaging (MRI) serves as the primary diagnostic tool but lacks specificity. Histopathological examination remains the gold standard for definitive diagnosis. The review underscores the importance of timely biopsy in suspected cases to facilitate accurate diagnosis and appropriate management. Management and Prognosis: Current management involves biopsy and chemotherapy; however, optimal treatment strategies remain ambiguous due to the rarity of PCEL. Despite aggressive therapeutic interventions, prognosis remains poor, emphasizing the urgency for enhanced diagnostic and treatment modalities. Conclusions: Primary cauda equina lymphoma poses diagnostic and therapeutic challenges, necessitating a high index of suspicion in patients with atypical spinal cord symptoms. Collaborative efforts between neurosurgical, oncological, and infectious diseases teams are imperative for timely diagnosis and management. Advancements in diagnostic precision and therapeutic options are crucial for improving patient outcomes.

Three-dimensional (3D) spheroid models aim to bridge the gap between traditional two-dimensional (2D) cultures and the complex in vivo tissue environment. These models, created by self-clustering cells to mimic a 3D environment with surrounding extracellular framework, provide a valuable research tool. The NSC-34 cell line, generated by fusing mouse spinal cord motor neurons and neuroblastoma cells, is essential for studying neurodegenerative diseases like amyotrophic lateral sclerosis (ALS), where abnormal protein accumulation, such as TAR-DNA-binding protein 43 (TDP-43), occurs in affected nerve cells. However, NSC-34 behavior in a 3D context remains underexplored, and this study represents the first attempt to create a 3D model to determine its suitability for studying pathology. We generated NSC-34 spheroids using a nonadhesive hydrogel-based template and characterized them for 6 days. Light microscopy revealed that NSC-34 cells in 3D maintained high viability, a distinct round shape, and forming stable membrane connections. Scanning electron microscopy identified multiple tunnel-like structures, while ultrastructural analysis highlighted nuclear bending and mitochondria alterations. Using inducible GFP-TDP-43-expressing NSC-34 spheroids, we explored whether 3D structure affected TDP-43 expression, localization, and aggregation. Spheroids displayed nuclear GFP-TDP-43 expression, albeit at a reduced level compared with 2D cultures and generated both TDP-35 fragments and TDP-43 aggregates. This study sheds light on the distinctive behavior of NSC-34 in 3D culture, suggesting caution in the use of the 3D model for ALS or TDP-43 pathologies. Yet, it underscores the spheroids' potential for investigating fundamental cellular mechanisms, cell adaptation in a 3D context, future bioreactor applications, and drug penetration studies.

Several theories have tried to elucidate the mechanisms behind the pathophysiology of chronic subdural hematoma (CSDH). However, this process is complex and remains mostly unknown. In this study we performed a retrospective randomised analysis comparing the cortical atrophy of 190 patients with unilateral CSDH, with 190 healthy controls. To evaluate the extent of cortical atrophy, CT scan images were utilised to develop an index that is the ratio of the maximum diameter sum of 3 cisterns divided by the maximum diameter of the skull at the temporal lobe level. Also, we reported, for the first time, the ultrastructural analyses of the CSDH using a combination of immunohistochemistry methods and transmission electron microscopy techniques. Internal validation was performed to confirm the assessment of the different degrees of cortical atrophy. Relative Cortical Atrophy Index (RCA index) refers to the sum of the maximum diameter of three cisterns (insular cistern, longitudinal cerebral fissure and cerebral sulci greatest) with the temporal bones' greatest internal distance. This index, strongly related to age in healthy controls, is positively correlated to the preoperative and post-operative maximum diameter of hematoma and the midline shift in CSDH patients. On the contrary, it negatively correlates to the Karnofsky Performance Status (KPS). The Area Under the Receiver Operating Characteristics (AUROC) showed that RCA index effectively differentiated cases from controls. Immunohistochemistry analysis showed that the newly formed CD-31 positive microvessels are higher in number than the CD34-positive microvessels in the CSDH inner membrane than in the outer membrane. Ultrastructural observations highlight the presence of a chronic inflammatory state mainly in the CSDH inner membrane. Integrating these results, we have obtained an etiopathogenetic model of CSDH. Cortical atrophy appears to be the triggering factor activating the cascade of transendothelial cellular filtration, inflammation, membrane formation and neovascularisation leading to the CSDH formation.

Glioblastoma is an aggressive brain tumor with an average life expectancy between 14 and 16 months after diagnosis. The Ki-67 labeling index (LI), a measure of cellular proliferation, is emerging as a prognostic marker in GBM. In this study, we investigated the ultrastructure of glioblastoma tissue from 9 patients with the same molecular profile (adult IDH wild-type glioblastoma, wild-type ATRX, and positive for TP53 expression, GFAP expression, and EGFR overexpression) to find possible ultrastructural features to be used as biomarkers and correlated with the only parameter that differs among our samples, the Ki-67 LI. Our main results were the visualization of the anatomical basis of astrocyte-endothelial cells crosstalk; the ultrastructural in situ imaging of clusters of hyperactivated microglia cells (MsEVs); the ultrastructural in situ imaging of microglia cells storing lipid vesicles (MsLVs); the ultrastructural in situ imaging of neoplastic cells mitophagy (NCsM). The statistical analysis of our data indicated that MsEVs and MsLVs correlate with the Ki-67 LI value. We can thus assume they are good candidates to be considered morphological biomarkers correlating to Ki-67 LI. The role of NCsM instead must be further evaluated. Our study findings demonstrate that by combining ultrastructural characteristics with molecular information, we can discover biomarkers that have the potential to enhance diagnostic precision, aid in treatment decision-making, identify targets for therapy, and enable personalized treatment plans tailored to each patient. However, further research with larger sample sizes is needed to validate these findings and fully utilize the potential of ultrastructural analysis in managing glioblastoma.

Although therapeutic targets have been developed for colorectal cancer (CRC) therapy, the therapeutic effects are not ideal and the survival rate for CRC patients remains poor. Therefore, it is crucial to recognize a specific target and develop an efficacious delivery system for CRC therapy. Herein, we demonstrate that reduced ALKBH5 mediates aberrant m6A modification and tumor progression in CRC. Mechanically, histone deacetylase 2-mediated H3K27 deacetylation inhibits ALKBH5 transcription in CRC, whereas ectopic ALKBH5 expression decreases tumorigenesis of CRC cells and protects mice from colitis-associated tumor development. Further, METTL14/ALKBH5/IGF2BPs combine to modulate JMJD8 stability in an m6A-dependent manner, which increases glycolysis and accelerates the development of CRC by enhancing the enzymatic activity of PKM2. Moreover, ALKBH5 mRNA-loaded folic acid-modified exosome-liposome hybrid nanoparticles were synthesized and significantly inhibit the progression of CRC in preclinical tumor models by modulating the ALKBH5/JMJD8/PKM2 axis and inhibiting glycolysis. Overall, our research confirms the crucial function of ALKBH5 in regulating the m6A status in CRC and provides a direct preclinical approach for using ALKBH5 mRNA nanotherapeutics for CRC.

The Locomotor System Anatomy (LSA) course, placed in the first semester of the first year of the new Master’s degree in Medicine and Surgery High Technology (MSHT) at the Sapienza University of Rome, was integrated with surgical and bioengineering content. This study investigated the educational value and the students’ perceptions of the effectiveness of these two types of integration, comparing surgical integration (SI) with engineering integration (EI). Anatomy knowledge and students’ opinions attending the LSA course in MSHT degree (n = 30) were compared with those of students (n = 32) attending another medical and surgery course not comprising EI. Data show that students in the MSHT course like in-depth SI much more than in-depth EI. However, those who like in-depth SI also like in-depth EI. Significant differences were in anatomy knowledge between the two groups in the three sections of the test. There was no significant correlation between the three test scores and the levels of liking, while there was a significant correlation between students liking SI and those liking EI. A statistically significant correlation was also found in students who correctly responded to questions on the head and trunk, with students responding correctly to questions on the upper limbs. This study will be important in optimizing the deepening of SI and EI in the LSA course.