Magnetic and pH-responsive magnetite/chitosan (core/shell) nanoparticles for dual-targeted methotrexate delivery in cancer therapy.

Methotrexate successful therapy encounters various challenges in chemotherapy, such as poor oral bioavailability, low specificity, side effects and the development of drug resistances. In this study, it is proposed a dual-targeted nanocarrier comprising magnetite/chitosan nanoparticles for an efficient Methotrexate delivery. The formation of the particles was confirmed through morphological analysis using electron microscopy and elemental mappings via energy dispersive X-ray spectroscopy. These nanoparticles exhibited a size of ≈ 270 nm, a zeta potential of ≈ 24 mV, and magnetic responsiveness, as demonstrated by hysteresis cycle analysis and visual observations under a magnetic field. In addition, these particles displayed high stability, as evidenced by size and surface electric charge measurements, during storage at both 4 ºC and 25 ºC for at least 30 days. Electrophoretic properties were examined in relation to pH and ionic strength, confirming these core/shell nanostructure. The nanoparticles demonstrated a pH-responsive drug release as observed by a sustained Methotrexate release over the next 90 h under pH ≈ 7.4, while complete release occurred within 3 h under acidic conditions (pH ≈ 5.5). In the biocompatibility assessment, the magnetite/chitosan particles showed excellent hemocompatibility ex vivo and no cytotoxic effects on normal MCF-10 A and cancer MCF-7 cells. Furthermore, the Methotrexate-loaded nanoparticles significantly enhanced the antitumor activity reducing the half-maximal inhibitory concentration by ≈ 2.7-fold less compared to the free chemotherapeutic.

Mitigating flood risk and environmental change in show caves: Key challenges in the management of the Las Güixas cave (Pyrenees, Spain).

A successful management of a show cave requires knowledge of cave dynamics and the main risk factors. Show caves close to the water table are prone to sporadic flooding, which can threaten visitor safety and result in severe economic losses. Las Güixas cave, located in the Collarada Massif (Pyrenees - Spain), is representative of a show cave close to the water table that is exposed to energetic flash floods. We conducted a five-year comprehensive cave monitoring study including air temperature, relative humidity, CO2 concentration and water level. Additionally, we measured outside temperature and precipitation. Air temperature variations and ventilation dynamics occurring in most of the cave are controlled by the outside temperature due to entrances at different elevations, except in a non-ventilated area showing more stable hygrothermal characteristics and higher summer values of CO2 concentration. The study also identifies distinct CO2 sources related to the degassing of water and visitors' breathing. Monitoring data show that the rapid degassing of cave water during flooding may increase subsurface CO2 concentrations to levels well above the exposure limits. However, the strong ventilation observed inside the cave rapidly removes CO2 peaks produced by flooding and limits the anthropic CO2 rise to ∼100 ppm. Hydrograph analysis revealed a response time of 8-12 h in the cave water levels to external rainfall/snowmelt events. Based on these results, a flood alarm system supports sustainable show cave management and the number of visitors is optimized according to the environmental conditions of the cave. This monitoring study has greatly contributed to our knowledge of cave dynamics, which can serve to improve flood risk management and increase the profitability of the show cave. Nonetheless, extreme floods remain a significant concern for potential economic losses in the future, considering current climate change scenarios. Hydrological studies together with a long-term monitoring will allow evaluating the impact of future changes in climate and environmental parameters.

"Skeletal surgical approach in Down Syndrome with Obstructive Sleep Apnea."

Failing to address Obstructive Sleep Apnea (OSA) in Down Syndrome (DS) patients can have serious consequences, leading to increased mortality. Moreover, a notable portion of individuals find it challenging to tolerate Continuous Positive Airway Pressure (CPAP). Therefore, this study aims to share our experiences in treating adult DS patients with moderate to severe OSA who don't tolerate CPAP using various surgical approaches. A retrospective analytical study including 20 DS with moderate to severe/very severe OSA who had no tolerance to Continuous Positive Airway Pressure (CPAP) was conducted. Regarding the individual skeletal characteristics of each patient various orthognathic surgery techniques were performed. Two in-hospital polysomnographies (PSG) were undertaken for every patient; one before the surgical procedure, while another was performed between 6 and 60 months of the follow-up post-surgery period. Polysomnographic variables were analyzed in this study such as AHI, ODI, and T90, which all showed statistically significant improvement after surgery with a p value in the Wilcoxon test <0,01. These results have remained stable over time, with no recurrences of OSA observed over the five years of follow-up. Thus, orthognathic surgery may be the only viable option for individuals with DS who are unable to tolerate CPAP.

Insights on the Mechanical Properties of SARS-CoV-2 Particles and the Effects of the Photosensitizer Hypericin.

SARS-CoV-2 is a highly pathogenic virus responsible for the COVID-19 disease. It belongs to the Coronaviridae family, characterized by a phospholipid envelope, which is crucial for viral entry and replication in host cells. Hypericin, a lipophilic, naturally occurring photosensitizer, was reported to effectively inactivate enveloped viruses, including SARS-CoV-2, upon light irradiation. In addition to its photodynamic activity, Hyp was found to exert an antiviral action also in the dark. This study explores the mechanical properties of heat-inactivated SARS-CoV-2 viral particles using Atomic Force Microscopy (AFM). Results reveal a flexible structure under external stress, potentially contributing to the virus pathogenicity. Although the fixation protocol causes damage to some particles, correlation with fluorescence demonstrates colocalization of partially degraded virions with their genome. The impact of hypericin on the mechanical properties of the virus was assessed and found particularly relevant in dark conditions. These preliminary results suggest that hypericin can affect the mechanical properties of the viral envelope, an effect that warrants further investigation in the context of antiviral therapies.

Development of two competitive ELISAs based on monoclonal antibodies for the serological detection of Bovine ephemeral fever virus.

Bovine ephemeral fever virus (BEFV) is a member of the genus Ephemerovirus in the family Rhabdoviridae. It is an arthropod-borne virus transmitted by many species of midges and mosquitoes. It can cause severe economic consequences due to losses in milk production and the general condition of cattle and water buffalo. BEF occurs in some tropical, subtropical and warm temperate regions of Africa, Australia, the Middle East and Asia with seasonal outbreaks, but its possible spread to other areas (e.g. Europe) cannot be excluded. Therefore, using and developing rapid diagnostic methods with optimal performance is essential for identifying emerging pathogens and their control. In the present study, we developed two competitive serological ELISAs based on monoclonal antibodies (mAbs), designed by using BEFV inactivated antigen and the BEF recombinant nucleoprotein (N), respectively. A panel of 77 BEF-positive and 338 BEF-negative sera was used to evaluate the two tests. With a diagnostic sensitivity of 97.4 % using the inactivated virus and 98.7 % using the recombinant N, and a diagnostic specificity of 100 % using both antigens, our results suggest that these tests are suitable for the serological diagnosis of BEF.

Revisiting plant cuticle biophysics.

The plant cuticle is located at the interface of the plant with the environment, thus acting as a protective barrier against biotic and abiotic external stress factors, and regulating water loss. Additionally, it modulates mechanical stresses derived from internal tissues and also from the environment. Recent advances in the understanding of the hydric, mechanical, thermal, and, to a lower extent, optical and electric properties of the cuticle, as well as their phenomenological connections and relationships are reviewed. An equilibrium based on the interaction among the different biophysical properties is essential to ensure plant growth and development. The notable variability reported in cuticle geometry, surface topography, and microchemistry affects the analysis of some biophysical properties of the cuticle. This review aimed to provide an updated view of the plant cuticle, understood as a modification of the cell wall, in order to establish the state-of-the-art biophysics of the plant cuticle, and to serve as an inspiration for future research in the field.

Screening system for the detection of unknown or poorly controlled diabetes mellitus in a tertiary hospital: A retrospective study.

RATIONALE: Hospitalization may be a valuable chance for the detection of unknown and uncontrolled diabetes mellitus (DM). There is a screening tool at our hospital: in case of high inpatient plasma glucose level, an A1c value is added if no available from the last 3 months. AIMS AND OBJECTIVES: Our objective was to analyse the population with A1c ≥ 9% detected through this system from 2021 to 2023. MATERIALS AND METHODS: A retrospective study was performed. Three thousand five hundred seventy-two patients were screened. We studied 243 patients with A1c ≥ 9%. RESULTS: Fourty-eight patients (19.8%) had unknown DM. The Endocrinology department was consulted in 39 cases (16%). In most of the cases (51%), there was not a mention in the discharge report, nor changes in the usual treatment (65.4%). Ninety patients (37%) improved A1c. Most of the ones that improved (58.9%) had a correct follow-up, compared with those who did not (23.5%) (p < 0.01). CONCLUSIONS: Measurement of A1c during hospitalization can help us to diagnose unknown or poorly controlled DM. Therapeutic inertia and delayed diagnosis are two problems associated to DM that are confirmed by our data.

Isolation and Molecular Evidence of Tunisian Sheep-like Pestivirus (Pestivirus N) in Persistently Infected Sheep in Northern Italy, 2023.

Over the last few decades, several pestiviruses have been discovered in ruminants, pigs, and, more recently, in non-ungulate hosts. Consequently, the nomenclature and taxonomy of pestiviruses have been updated. The Tunisian sheep-like pestivirus (TSV, Pestivirus N) is an additional ovine pestivirus genetically closely related to classical swine fever virus (CSFV). In this study, during a survey of pestivirus infections in ovine farms in the Lombardy region of Northern Italy, we identified and isolated a pestivirus strain from a sheep that was found to belong to Pestivirus N species based on its genomic nucleotide identity. The sheep itself and its lamb were found to be persistently infected. We performed molecular characterization and phylogenetic analysis of three viral genomic regions (a fragment of 5'-UTR, partial Npro, and the whole E2 region). In conclusion, these results confirmed circulating TSV in Northern Italy after notification in Sicily, Italy, and France. Correlation with Italian, Tunisian, and French strains showed that detection might have resulted from the trading of live animals between countries, which supports the need for health control measures.

IL-17 signaling in primary sclerosing cholangitis patient-derived organoids.

BACKGROUND: The pathogenesis of primary sclerosing cholangitis (PSC) is unclear, although studies implicate IL-17A as an inflammatory mediator in this disease. However, a direct assessment of IL-17 signaling in PSC cholangiocytes is lacking. In this study, we aimed to investigate and characterize the response of PSC extrahepatic cholangiocyte organoids (ECO) to IL-17A stimulation. METHODS: Cholangiocytes obtained from patients with PSC and without PSC by endoscopic retrograde cholangiography were cultured as ECO. The ECO were treated with vehicle or IL-17A and assessed by transcriptomics, secretome analysis, and genome sequencing. RESULTS: Unsupervised clustering of all integrated single-cell RNA sequencing data identified 8 cholangiocyte clusters that did not differ between PSC and non-PSC ECO. However, PSC ECO cells demonstrated a robust response to IL-17 treatment, as noted by an increased number of differentially expressed genes by transcriptomics and more abundant chemokine and cytokine expression and secretion. After rigorous filtering, genome sequencing identified candidate somatic variants shared among PSC ECO from unrelated individuals. However, no candidate rare variants in genes regulating the IL-17 pathway were identified, but rare variants regulating the MAPK signaling pathway were present in all PSC ECO. CONCLUSIONS: PSC and non-PSC patient-derived ECO respond differently to IL-17 stimulation, implicating this pathway in the pathogenesis of PSC.

Clinical Interpretation of Genetic Variants in the Evaluation and Management of Thoracic Aortic Aneurysm and Dissection.

BACKGROUND: We aimed to elucidate clinical implications of genetic variant interpretation in assessing disease severity and progression in thoracic aortic aneurysm and dissection (TAAD) patients. METHODS: Consecutive TAAD patients with aortic root and/or ascending aortic aneurysms seen between 2011 and 2020 were included. Serial echocardiography, family history of TAAD, and management information were retrospectively collected and analyzed. Patients were classified into gene-positive (Gen-P), variants of uncertain significance, and gene-negative (Gen-N) groups. RESULTS: A total of 407 patients were included: mean age 53.7 ± 15.4 years, 64.4% men, and 38% with reported family history of TAAD. Thirty-seven (9.1%) were Gen-P; 147 (36.1%) had a variant of uncertain significance. The maximal aneurysm diameter was 4.78 mm larger in Gen-P than the other groups (P < .001). In 162 unoperated TAAD patients with serial echocardiographic measurements, aneurysms enlarged at a significantly higher rate in the Gen-P (1.36 mm/year, 95% CI: 0.77-1.95) than variants of uncertain significance and Gen-N groups (0.83 mm/year vs 0.89 mm/year, respectively; P < .001). Aneurysms were 20% more likely to require surgical intervention for every millimeter increase in diameter. When considered on an individual basis, the highest growth rates were found in the variants of uncertain significance group. CONCLUSIONS: While aneurysms linked to variants of uncertain significance demonstrate average growth rates comparable to those in Gen-N, close follow-up and genetic counseling in the variants of uncertain significance group are recommended for assessment of pathogenicity on a case-by-case basis. Early familial gene testing in TAAD is important to develop individualized preventive and therapeutic criteria.

Antibacterial Activity of Epigallocatechin-3-gallate (EGCG) Loaded Lipid-chitosan Hybrid Nanoparticle against Planktonic Microorganisms.

Epigallocatechin-3-gallate (EGCG), a polyphenol derived from Green Tea, is one of the sources of natural bioactive compounds which are currently being developed as medicinal ingredients. Besides other biological activities, this natural compound exhibits anti-cariogenic effects. However, EGCG has low physical-chemical stability and poor bioavailability. Thus, the purpose of this study was to develop and characterize lipid-chitosan hybrid nanoparticle with EGCG and to evaluate its in vitro activity against cariogenic planktonic microorganisms. Lipid-chitosan hybrid nanoparticle (LCHNP-EGCG) were prepared by emulsion and sonication method in one step and characterized according to diameter, polydispersity index (PdI), zeta potential (ZP), encapsulation efficiency (EE), mucoadhesion capacity and morphology. Strains of Streptococcus mutans, Streptococcus sobrinus and Lactobacillus casei were treated with LCHNP- EGCG, and minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were evaluated. LCHNP-EGCG exhibited a size of 217.3 ± 5.1 nm with a low polydispersity index (0.17) and positive zeta potential indicating the presence of chitosan on the lipid nanoparticle surface (+33.7 mV). The LCHNP-EGCG showed a spherical morphology, high stability and a mucoadhesive property due to the presence of chitosan coating. In addition, the EGCG encapsulation efficiency was 96%. A reduction of almost 15-fold in the MIC and MBC against the strains was observed when EGCG was encapsulated in LCHNP, indicating the potential of EGCG encapsulation in lipid-polymer hybrid nanoparticles. Taking the results together, the LCHNP-EGCG could be an interesting system to use in dental care due to their nanometric size, mucoadhesive properties high antibacterial activity against relevant planktonic microorganisms.

Using "Adopt a Bacterium" as an e-learning tool for simultaneously teaching microbiology to different health-related university courses.

The COVID-19 pandemic has posed challenges for education, particularly in undergraduate teaching. In this study, we report on the experience of how a private university successfully addressed this challenge through an active methodology applied to a microbiology discipline offered remotely to students from various health-related courses (veterinary, physiotherapy, nursing, biomedicine, and nutrition). Remote teaching was combined with the "Adopt a Bacterium" methodology, implemented for the first time on Google Sites. The distance learning activity notably improved student participation in microbiology discussions, both through word cloud analysis and the richness of discourse measured by the Shannon index. Furthermore, feedback from students about the e-learning approach was highly positive, indicating its effectiveness in motivating and involving students in the learning process. The results also demonstrate that despite being offered simultaneously to students, the methodology allowed for the acquisition of specialized knowledge within each course and sparked student interest in various aspects of microbiology. In conclusion, the remote "Adopt a Bacterium" methodology facilitated knowledge sharing among undergraduate students from different health-related courses and represented a valuable resource in distance microbiology education.

Immunomodulatory potential of rapamycin-loaded mesoporous silica nanoparticles: pore size-dependent drug loading, release, and in vitro cellular responses.

Rapamycin is a potent immunosuppressive drug that has been recently proposed for a wide range of applications beyond its current clinical use. For some of these proposed applications, encapsulation in nanoparticles is key to ensure therapeutic efficacy and safety. In this work, we evaluate the effect of pore size on mesoporous silica nanoparticles (MSN) as rapamycin nanocarriers. The successful preparation of MSN with 4 different pore sizes was confirmed by dynamic light scattering, zeta potential, transmission electron microscopy and N2 adsorption. In these materials, rapamycin loading was pore size-dependent, with smaller pore MSN exhibiting greater loading capacity. Release studies showed sustained drug release from all MSN types, with larger pore MSN presenting faster release kinetics. In vitro experiments using the murine dendritic cell (DC) line model DC2.4 showed that pore size influenced the biological performance of MSN. MSN with smaller pore sizes presented larger nanoparticle uptake by DC2.4 cells, but were also associated with slightly larger cytotoxicity. Further evaluation of DC2.4 cells incubated with rapamycin-loaded MSN also demonstrated a significant effect of MSN pore size on their immunological response. Notably, the combination of rapamycin-loaded MSN with an inflammatory stimulus (lipopolysaccharide, LPS) led to changes in the expression of DC activation markers (CD40 and CD83) and in the production of the proinflammatory cytokine TNF-α compared to LPS-treated DC without nanoparticles. Smaller-pored MSN induced more substantial reductions in CD40 expression while eliciting increased CD83 expression, indicating potential immunomodulatory effects. These findings highlight the critical role of MSN pore size in modulating rapamycin loading, release kinetics, cellular uptake, and subsequent immunomodulatory responses.

Monitoring of Home Respiratory Polygraphy During Mandibular Distraction Osteogenesis Succeeded by LeFort I Osteotomy in Adult Patients With Obstructive Sleep Apnea.

BACKGROUND: Bilateral internal ramus distraction (BIRD) is now part of select protocols for treating obstructive sleep apnea (OSA). Introducing a home monitoring protocol offers a valuable alternative to overnight laboratory polysomnography (PSG). PURPOSE: The purpose of this study was to evaluate and compare OSA parameters obtained via home respiratory polygraphy (HRP) and hospital PSG in a cohort of patients undergoing mandibular distraction for OSA management. STUDY DESIGN, SETTING, SAMPLE: Hospital Universitario La Princesa (Madrid) researchers conducted a prospective cohort study with patients diagnosed with moderate (apnea-hypopnea index (AHI) = 15 to 30) to severe (AHI>30) OSA undergoing BIRD followed by LeFort maxillary osteotomy. Exclusion criteria were as follows: severe systemic diseases, central apneas, smoking, poor dental hygiene, or prior OSA interventions. PREDICTOR VARIABLE: The predictor variable was PSG and HRP techniques. MAIN OUTCOME VARIABLES: The main outcome variable comprises a collection of OSA parameters, including the AHI, oxygen desaturation index (ODI), and time spent below 90% oxygen saturation (T90). These data were measured both before and after distraction, as well as after 6 mm, 9 mm, and 12 mm of mandibular distraction. COVARIATES: The covariates were age, sex, cardiovascular risk parameters, and the Epworth sleepiness scale. ANALYSES: Pearson's correlation analyzed AHI, ODI, and T90 values from PSG and HRP. Wilcoxon Signed Rank-Sum Test compared 2 distraction stages, and the Friedman Test evaluated 3 stages (P < .05). Multiple regression analysis assessed if covariates were independent risk factors for postoperative persistent OSA. RESULTS: The study included 32 patients (25% with moderate and 75% with severe OSA). Final AHI was 10.9 ± 8.9 (events/hour) with HRP, compared to 15.2 ± 13.4 with PSG (r = 0.7, P < .05). ODI was 9.0 ± 8.1 (des/h) with HRP and 8.7 ± 9.5 with PSG (r = 0.85, P < .05). T90 was 1.6 ± 2.2 with HRP and 1.3 ± 3.0 with PSG (r = 0.6, P < .05). Based on HRP data, AHI improved from D1 (34.0 ± 19.5) to D2 (20.8 ± 14.1) and D3 (12.5 ± 10.4) (P < .05). ODI decreased from D1 (26.1 ± 19.0) to D2 (16.0 ± 12.6) and D3 (9.4 ± 8.8) (P < .05). T90 reduced from D1 (8.2 ± 12.6) to D2 (4.1 ± 5.2) and D3 (1.9 ± 2.8) (P < .05). Multiple regression analysis of comorbidities produced nonsignificant results. CONCLUSION AND RELEVANCE: Monitoring BIRD through HRP has demonstrated efficacy in yielding results that align with PSG.

Detection of Influenza D Antibodies in Dogs, Apulia Region, Italy, 2016 and 2023.

Dogs are known to be susceptible to influenza A viruses, although information on influenza D virus (IDV) is limited. We investigated the seroprevalence of IDV in 426 dogs in the Apulia region of Italy during 2016 and 2023. A total of 14 samples were positive for IDV antibodies, suggesting exposure to IDV in dogs.

Molecular and Serological Detection of Bovine Coronaviruses in Marmots (Marmota marmota) in the Alpine Region.

In this study, virological surveillance focused on coronaviruses in marmots in the Alpine region in 2022, captured as part of a population control reduction program in the Livigno area. Seventy-six faecal samples were randomly collected from marmots at the time of capture and release and tested for genome detection of pan-coronavirus, pan-pestivirus, canine distemper virus, and influenza A and D virus. Nine faecal samples were positive in the Pan-CoV RT-PCR, while all were negative for the other viruses. Pan-coronavirus positives were further identified using Illumina's complete genome sequencing, which showed the highest homology with Bovine Coronavirus previously detected in roe deer in the Alps. Blood samples (n.35) were collected randomly from animals at release and tested for bovine coronavirus (BCoV) antibodies using competitive ELISA and VNT. Serological analyses revealed that 8/35 sera were positive for BCoV antibodies in both serological tests. This study provides molecular and serological evidence of the presence of BCoV in an alpine marmot population due to a likely spillover event. Marmots share areas and pastures with roe deer and other wild ruminants, and environmental transmission is a concrete possibility.

Voided urine cytology is a useful tool predicting non-muscle-invasive bladder cancer risk before surgery.

OBJECTIVE: To determine the accuracy of voided urinary cytology (VUC) in predicting of non-muscle-invasive bladder cancer (NMIBC) risk stratification before surgery. METHODS: We prospectively collected data from all patients diagnosed with bladder cancer in our institution over 2 years. We have analyzed VUC accuracy of positive and suspicious VUC in the detection of high-risk tumors and negative and atypical VUC in the detection of low-risk tumors. To test this accuracy, we assessed sensitivity, specificity, positive (PPV) and negative predictive values (NPV), diagnostic odds ratio (DOR), and generated ROC curves (receiver operating characteristic curve). RESULTS: With 224 patients included, the positive VUC subcategory showed a specificity of 92.4% (95%CI: 83.2%-97.5%) and a PPV of 91.4 (95%CI: 81%-97.1%). DOR in this subgroup was 6.81. In the suspicious VUC, specificity was 90.9% (95%CI: 81.3%-96.6%), PPV was 88% (95%CI: 75.7%-95.5%) and DOR was 4.23. Combined analysis of positive and suspicious cytologies for detecting high-risk NMIBC showed a sensitivity of 65% (95%CI: 57.3%-73.2%) and a DOR of 9.51. Negative VUC showed high specificity in detecting low-risk (93.2% [95%CI: 87.9%-96.7%]) and a DOR of 6.90 (95%CI: 3.07-15.46). Atypical VUC was the least accurate and had rather low specificity and predictive values. CONCLUSIONS: VUC appears to be a good, inexpensive and easily available method to determine risk stratification before surgery. This can be useful in daily practice to determine which patients should receive a single instillation of MMC and to prioritize patients more likely to have a high- risk tumor.