RESUMEN

Accelerated material development for refractory ceramics triggers possibilities in context to enhanced energy efficiency for industrial processes. Here, the gathering of comprehensive material data is essential. High temperature-confocal laser scanning microscopy (HT-CLSM) displays a highly suitable in-situ method to study the underlying dissolution kinetics in the slag over time. A major drawback concerns the efficient and accurate processing of the collected image data. Here, we introduce an attention encoder-decoder convolutional neural network enabling the fully automated evaluation of the particle dissolution rate with a precision of 99.1%. The presented approach provides accurate and efficient analysis capabilities with high statistical gain and is highly resilient to image quality changes. The prediction model allows an automated diameter evaluation of the MgO particles' dissolution in the silicate slag for different temperature settings and various HT-CLSM data sets. Moreover, it is not limited to HT-CLSM image data and can be applied to various domains.

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Objective: The objective of this study was to evaluate the effect of blood and artificial salivary contamination of different root-end filling materials on microleakage using a confocal laser scanning microscope and on marginal adaptation using a scanning electron microscope. Materials and Methods: Eighty noncarious single-rooted teeth with mature apices were taken. After retro-cavity preparation, they were randomly assigned into two major groups (n = 40). They were contaminated with blood and artificial saliva, respectively. Each major group was divided randomly into four subgroups (n = 10) and filled as follows: Subgroup A, Biodentine; Subgroup B, bioactive bone cement; Subgroup C, Cention N; and Subgroup D, Bio-C Repair. The samples were sectioned transversely at 1 and 2 mm from the root apex and checked under a confocal laser scanning microscope for microleakage and under an scanning electron microscope for marginal adaptation. The average mean values were calculated. Independent samples t-tests, paired t-tests, and one-way analysis of variance with Tukey's post hoc tests were done to analyze the data. Results: All the tested materials showed marginal gaps and dye leakage. The Bio-C Repair group showed the least mean marginal gap and dye leakage values, followed by bioactive bone cement, Biodentine, and Cention N, respectively, in both blood and artificial saliva contamination. However, the mean marginal gaps and dye leakage between the major groups were statistically insignificant. Conclusion: In an overall comparison, Bio-C Repair was found to be superior in terms of marginal adaptation and sealing ability under the test conditions.

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The excessive production and use of plastics increase the release of micro- and nanoplastics (MNPs) into the environment. In recent years, research has focused on the occurrence of MNPs in air, soil and water. Nevertheless, there is still a lack of knowledge regarding MNPs in plants. To determine the load, translocation of MNPs and their effects on metabolism, pak choi, tomato, radish and asparagus have been exposed with fluorescent-labelled poly(methyl methacrylate) or polystyrene (PS) MNPs. The entry of nanoparticles (NPs) of various sizes (100-500 nm) and surface modifications (unmodified, COOH or NH2) into plants has been demonstrated using confocal laser scanning microscopy (CLSM). The translocalization from root to shoot and the accumulation of NP in the intercellular spaces were regardless of the surface modification. In addition, metabolomics was used to evaluate metabolic changes induced by MNPs in pak choi. Changes in phenolic compounds, phytohormone derivatives and other classes of compounds known to be triggered by various environmental stresses have been identified. The present study demonstrates the uptake and translocalization of MNPs in edible parts of vegetables and may pose a hazard for humans.

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Aerosol whipped cream, widely used in various foods, relies on solid fats rich in saturated fatty acids (SFAs) for stable gas entrapment. In this study, the potential of oleogels as a healthy fat substitute for formulating aerosol whipped cream was studied. The analysis focused on the effects of different types (beeswax [BW], rice bran wax [RW], and carnauba wax) and the quantities of wax on the properties of the aerosol whipped creams. The BW-oleogel-based aerosol whipped cream exhibited the highest foam ability and foam stability. The superior physical properties of the bees wax-oleogel were attributed to the higher overrun and height stability of its whipped cream compared to RW and carnauba oleogels. The 6% BW-whipped cream showed significantly higher overrun and cream stability. The 6% BW oleogel whipped cream contained 4.4 times lower SFAs than the dairy milk fat whipped cream. This study represents the first exploration into the feasibility of formulating oleogel-based aerosol whipped cream using liquid vegetable oil.

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Membrane surface fouling has always been a critical issue for the long-term operation of polymeric membranes. Therefore, it is crucial to develop new approaches to prevent fouling. While developing new approaches, characterization methods are greatly important for understanding the distribution of fouling on the membrane surface. In this work, a cellulose acetate membrane was fouled by the filtration of artificial wastewater based on alginate. The surfaces of fouled membranes were characterized through scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), atomic force microscopy (AFM), and white light interferometry (WLI). The results were then compared in terms of the resolution, accuracy, feasibility, and cost-efficiency.

RESUMEN

Bacillus cereus is a well-known foodborne pathogen that can cause human diseases, including vomiting caused by emetic toxin, cereulide, requiring 105-108 cells per gram to cause the disease. The bacterial cells may be eliminated during processing, but cereulide can survive in most processing techniques due to its resistance to high temperatures, extreme pH and proteolytic enzymes. Herein, we reported dynamic processes of biofilm formation of four different types and cereulide production within the biofilm. Confocal laser scanning microscopy (CLSM) images revealed that biofilms of the four different types reach each stage at different time points. Among the extracellular polymeric substances (EPS) components of the four biofilms formed by the emetic B. cereus F4810/72 strain, proteins account for the majority. In addition, there are significant differences (p < 0.05) in the EPS components at the same stage among biofilms of different types. The time point at which cereulide was first detected in the four types of biofilms was 24 h. In the biofilm of B. cereus formed in ultra-high-temperature (UHT) milk, the first peak of cereulide appeared at 72 h. The cereulide content of the biofilms formed in BHI was mostly higher than that of the biofilms formed in UHT milk. This study contributes to a better understanding of food safety issues in the industry caused by biofilm and cereulide toxin produced by B. cereus.

Asunto(s)

Bacillus cereus , Biopelículas , Depsipéptidos , Microbiología de Alimentos , Bacillus cereus/metabolismo , Bacillus cereus/fisiología , Biopelículas/crecimiento & desarrollo , Depsipéptidos/metabolismo , Microscopía Confocal , Animales , Leche/microbiología , Calor , Matriz Extracelular de Sustancias Poliméricas/metabolismo , Enfermedades Transmitidas por los Alimentos/microbiología , Manipulación de Alimentos/métodos

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PURPOSE OF REVIEW: Quantification of the morphology of osteocyte lacunae has become a powerful tool to investigate bone metabolism, pathologies and aging. This review will provide a brief overview of 2D and 3D imaging methods for the determination of lacunar shape, orientation, density, and volume. Deviations between 2D-based and 3D-based lacunar volume estimations are often not sufficiently addressed and may give rise to contradictory findings. Thus, the systematic error arising from 2D-based estimations of lacunar volume will be discussed, and an alternative calculation proposed. Further, standardized morphological parameters and best practices for sampling and segmentation are suggested. RECENT FINDINGS: We quantified the errors in reported estimation methods of lacunar volume based on 2D cross-sections, which increase with variations in lacunar orientation and histological cutting plane. The estimations of lacunar volume based on common practice in 2D imaging methods resulted in an underestimation of lacunar volume of up to 85% compared to actual lacunar volume in an artificial dataset. For a representative estimation of lacunar size and morphology based on 2D images, at least 400 lacunae should be assessed per sample.

Asunto(s)

Imagenología Tridimensional , Osteocitos , Humanos , Osteocitos/citología , Imagenología Tridimensional/métodos , Densidad Ósea , Huesos/diagnóstico por imagen , Huesos/patología , Procesamiento de Imagen Asistido por Computador/métodos

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Performing accurate Fluorescence Correlation Spectroscopy (FCS) measurements in cells can be challenging due to cellular motion or other intracellular processes. In this respect, it has recently been shown that analysis of FCS data in short temporal segments (segmented FCS) can be very useful to increase the accuracy of FCS measurements inside cells. Here, we demonstrate that segmented FCS can be performed on a commercial laser scanning microscope (LSM), even in the absence of the dedicated FCS module. We show how data can be acquired on a Leica SP8 confocal microscope and then exported and processed with a custom software in MATLAB. The software performs segmentation of the data to extract an average ACF and measure the diffusion coefficient in specific subcellular regions. First of all, we measure the diffusion of fluorophores of different size in solution, to show that good-quality ACFs can be obtained in a commercial LSM. Next, we validate the method by measuring the diffusion coefficient of GFP in the nucleus of HeLa cells, exploiting variations of the intensity to distinguish between nucleoplasm and nucleolus. As expected, the measured diffusion coefficient of GFP is slower in the nucleolus relative to nucleoplasm. Finally, we apply the method to HeLa cells expressing a PARP1 chromobody to measure the diffusion coefficient of PARP1 in different subcellular regions. We find that PARP1 diffusion is slower in the nucleolus compared to the nucleoplasm.

Asunto(s)

Microscopía Confocal , Espectrometría de Fluorescencia , Humanos , Células HeLa , Microscopía Confocal/métodos , Espectrometría de Fluorescencia/métodos , Proteínas Fluorescentes Verdes/metabolismo , Difusión , Núcleo Celular/metabolismo , Programas Informáticos , Poli(ADP-Ribosa) Polimerasa-1/metabolismo

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Aim: The aim of the study was to evaluate and compare the depth of penetration of adhesive resin in resin-modified glass ionomer cement (RMGIC) after different surface pretreatment methods using a confocal laser scanning microscope (CLSM). Methods: Class I cavity preparation was done in 45 freshly extracted premolars and divided into three groups, according to the surface pretreatment of RMGIC which was placed in a thickness of 1 mm in the cavity. In the control group, no pretreatment was done, and other two groups were pretreated with acid etching and air abrasion (AA). The fifth-generation dentin bonding agent mixed with rhodamine B dye 0.1% was applied on RMGIC in all the samples and cured for 15 s. Samples were restored using composite resin and sectioned longitudinally. The depth of penetration of adhesive resin was evaluated using CLSM. Results: Depth of penetration of adhesive resin was highest with AA (153.70 ± 10.23), followed by acid etching (122.71 ± 12.25) and control group (77.12 ± 6.37). Conclusion: Based on the findings of this research, AA enhances the depth of penetration of adhesive resin in the RMGIC surface. Thus, AA before placement of composite resin on the RMGIC can be effective in a clinical scenario.

RESUMEN

This study aimed to assess root canal disinfection through various irrigation protocols, including a novel photothermal system called 'LEAP'. Mandibular premolars were infected with Enterococcus faecalis and divided into five groups for different treatments: Group 1: standard needle irrigation; Group 2: passive ultrasonic irrigation; Group 3: GentleWave; Group 4: LEAP; and Group 5: Group 1 + Group 4. Microbial counts were measured before (S1) and after disinfection (S2) using colony-forming units (CFU) and confocal laser scanning microscopy (CLSM). Results revealed a significant reduction in bacterial counts for all groups (p < 0.05). While the percentage of dead bacteria near the canal wall (0-50 µm) did not differ significantly, at 50-150 µm, LEAP and SNI + LEAP exhibited significantly higher bacterial reduction than other groups (p < 0.05). The findings indicate that LEAP is comparable to existing irrigation devices in the main root canal and notably superior in tubular disinfection.

RESUMEN

OBJECTIVE: Tolterodine tartrate (TOTA) is a first-line therapy to treat overactive urinary bladder (OAB). Oral delivery causes high hepatic clearance, xerostomia, headache, constipation, and blurred vision. We addressed Hansen solubility parameter (HSP) and Design Expert oriented optimized cationic elastic liposomes for transdermal application. METHODS: The experimental solubility was conducted in HSPiP predicted excipients to tailor formulations using surfactants, stearylamine, ethanol, and phosphatidylcholine (PC). These were evaluated for formulation characteristics. The optimized OTEL1 and OTEL1-G (gel) were compared against the drug solution (DS) and liposomes. In vitro and ex vivo studies were accomplished to investigate the insights into the mechanistic understanding of TOTA release and permeation ability. Finally, confocal laser scanning microscopy (CLSM) supported ex vivo results. RESULTS: HSP values of TOTA were closely related to tween-80, stearylamine, and human's skin. The size (153 nm), %EE (87.6%), and PDI (0.25) values of OTEL1 were in good agreement to the predicted values (161 nm, 80.4%, and 0.31) with high desirability (0.963). Spherical and smooth OTEL1 (including OTEL1-G and liposomes) vesicles followed non-Fickian drug release as compared to DS (Fickian) as evidence with n > 0.5 (Korsmeyer and Peppas coefficient). OTEL1 (containing lipid and surfactant as 90 mg and 13.8 mg, respectively) exhibited 2.6 and 1.8-folds higher permeation flux than DS and liposomes, respectively. Biocompatible cationic OTEL1 was safe and non-hemolytic. CONCLUSIONS: OTEL1 was promised as a lead vesicular approach and an alternative to conventional oral therapy to treat OAB in children and advanced age patients.

Asunto(s)

Administración Cutánea , Cationes , Liposomas , Absorción Cutánea , Solubilidad , Tartrato de Tolterodina , Humanos , Animales , Tartrato de Tolterodina/administración & dosificación , Tartrato de Tolterodina/farmacocinética , Cationes/química , Piel/metabolismo , Liberación de Fármacos , Excipientes/química , Masculino , Vejiga Urinaria Hiperactiva/tratamiento farmacológico , Composición de Medicamentos/métodos , Aminas

RESUMEN

In part I, we reported Hansen solubility parameters (HSP, HSPiP program), experimental solubility at varied temperatures for TOTA delivery. Here, we studied dose volume selection, stability, pH, osmolality, dispersion, clarity, and viscosity of the explored combinations (I-VI). Ex vivo permeation and deposition studies were performed to observe relative diffusion rate from the injected site in rat skin. Confocal laser scanning microscopy (CLSM) study was conducted to support ex vivo findings. Moreover, GastroPlus predicted in vivo parameters in humans and the impact of various critical factors on pharmacokinetic parameters (PK). Immediate release product (IR) contained 60% of PEG400 whereas controlled release formulation (CR) contained PEG400 (60%), water (10%) and d-limonene (30%) to deliver 2 mg of TOTA. GastroPlus predicted the plasma drug concentration of weakly basic TOTA as function of pH (from pH 2.0 to 9). The cumulative drug permeation and drug deposition were found to be in the order as B-VI˃ C-VI˃A-VI across rat skin. This finding was further supported with CLSM. Moreover, IR and CR were predicted to achieve Cmax of 0.0038 µg/ mL and 0.00023 µg/mL, respectively, after sub-Q delivery. Added limonene in CR extended the plasma drug concentration over period of 12 h as predicted in GastroPlus. Parameters sensitivity analysis (PSA) assessment predicted that sub-Q blood flow rate is the only factor affecting PK parameters in IR formulation whereas this was insignificant for CR. Thus, sub-Q delivery CR would be promising alternative with ease of delivery to children and aged patient.

Asunto(s)

Absorción Cutánea , Solubilidad , Tartrato de Tolterodina , Animales , Ratas , Humanos , Absorción Cutánea/efectos de los fármacos , Absorción Cutánea/fisiología , Tartrato de Tolterodina/administración & dosificación , Tartrato de Tolterodina/farmacocinética , Termodinámica , Solventes/química , Piel/metabolismo , Concentración de Iones de Hidrógeno , Preparaciones de Acción Retardada/farmacocinética , Preparaciones de Acción Retardada/administración & dosificación , Terpenos/química , Terpenos/administración & dosificación , Terpenos/farmacocinética , Administración Cutánea , Limoneno/administración & dosificación , Limoneno/farmacocinética , Limoneno/química , Masculino , Polietilenglicoles/química , Sistemas de Liberación de Medicamentos/métodos , Química Farmacéutica/métodos , Ciclohexenos/química , Ciclohexenos/farmacocinética , Ciclohexenos/administración & dosificación , Ratas Sprague-Dawley

RESUMEN

BACKGROUND: The present study aimed to evaluate the anti-hypertensive and anti-diabetic activities from biologically active peptides produced by fermented sheep milk with Lacticaseibacillus paracasei M11 (MG027695), as well as to purify and characterize the angiotensin-converting enzyme (ACE) inhibitory and anti-diabetic peptides produced from fermented sheep milk. RESULTS: After 48 h of fermentation at 37 °C, sheep milk demonstrated significant changes in anti-diabetic effects and ACE-I effects, with inhibition percentages observed for ACE inhibition (76.32%), α-amylase (70.13%), α-glucosidase (70.11%) and lipase inhibition (68.22%). The highest level of peptides (9.77 mg mL-1) was produced by optimizing the growth conditions, which included an inoculation rate of 2.5% and a 48 h of incubation period. The comparison of molecular weight distributions among protein fractions was conducted through sodium dodecyl-sulfate polyacrylamide gel electrophoresis analysis, whereas spots were separated using 2D gel electrophoresis according to both the molecular weight and pH. Peptide characterization with ultra-filtration membranes at 3 and 10 kDa allowed the study to assess molecular weight-based separation. Nitric oxide generated by lipopolysaccharide and the secretion of pro-inflammatory cytokines in RAW 264.7 immune cells were both inhibited by sheep milk fermented with M11. Fourier-transform infrared spectroscopy was employed to assess changes in functional groups after fermentation, providing insights into the structural changes occurring during fermentation. CONCLUSION: The present study demonstrates that fermentation with L. paracasei (M11) led to significant changes in fermented sheep milk, enhancing its bioactive properties, notably in terms of ACE inhibition and anti-diabetic activities, and the generation of peptides with bioactive properties has potential health benefits. © 2024 Society of Chemical Industry.

RESUMEN

AIM: The osteogenic potential of new premixed calcium-silicate-containing bioceramic sealers (Ca-Si sealers) was tested with porcine vascular wall-mesenchymal stem cells (pVW-MSCs). METHODOLOGY: Two Ca-Si-containing sealers: Ceraseal (MetaBiomed, Cheong-si, South Korea) and AH Plus Bioceramic (Maruchi, Wonju-si, South Korea), and an epoxy resin sealer (AH Plus; Dentsply, Konstanz, Germany) as a control, were prepared according to the manufacturers' indications. All samples were allowed to set for 100% of their setting time in a sterile humid cabinet at 37°C and 95% relative humidity. pVW-MSC seeding efficiency and osteogenic differentiation were analysed as marker of gene/protein expression for up to 12 days. Mineralization assay and immunofluorescence staining were performed and evaluated over a period of 21 days. Statistical analyses were conducted using one-way analysis of variance (p < .05). Additional samples were prepared and stored under the same conditions and inspected using an environmental scanning electron microscope equipped with an energy dispersive X-ray spectroscopy system. RESULTS: Significantly higher cell seeding efficiency (p < .05) was observed for both Ca-Si sealers from day 8. pVW-MSCs showed a significant shift towards the osteogenic lineage only when seeded in contact with Ca-Si sealers. Gene expression of osteopontin was upregulated significantly. Collagen I and osteocalcin were clearly expressed by cells in contact with Ca-Si sealers. Mineralization granules were observed in Alizarin red assays and confocal laser scanning microscopy analysis of both Ca-Si sealers. No gene expression or granule mineralization were observed on the epoxy resin sealer. CONCLUSIONS: Premixed Ca-Si sealers displayed a higher potential for osteogenic activity on pVW-MSCs. Epoxy resin sealer was unable to induce any osteogenic activity. The properties of both Ca-Si sealers suggest their potential as osteoinductive platforms for vascular MSCs in periapical bone.

Asunto(s)

Compuestos de Calcio , Células Madre Mesenquimatosas , Osteogénesis , Materiales de Obturación del Conducto Radicular , Silicatos , Células Madre Mesenquimatosas/efectos de los fármacos , Compuestos de Calcio/farmacología , Silicatos/farmacología , Animales , Materiales de Obturación del Conducto Radicular/farmacología , Osteogénesis/efectos de los fármacos , Porcinos , Diferenciación Celular/efectos de los fármacos , Cerámica/farmacología , Células Cultivadas , Materiales Biocompatibles/farmacología , Microscopía Electrónica de Rastreo , Ensayo de Materiales

RESUMEN

This study aimed to evaluate the efficacy of different irrigation techniques including standard needle irrigation (SNI), passive ultrasonic irrigation (PUI), EndoActivator (EA), XP-Endo Finisher (XPF), photon-induced photoacoustic streaming (PIPS), and shock wave enhanced emission photo-acoustic streaming (SWEEPS) systems on the penetration of irrigation solutions into dentinal tubules in teeth with calcium hydroxide (CH) applied and oval-shaped canals by confocal laser scanning microscope (CLSM). Ninety mandibular incisor teeth with oval-shaped canals were included in this study. After the preparation of teeth using Resiproc Blue R25 (VDW, Munich, Germany) canals were filled with a paste based on CH. The teeth were stored in 100% humidity at a temperature of 37°C for 14 days. According to the irrigation activation systems, the teeth were divided into six groups (n = 15); SNI, PUI, EA, XPF, PIPS, and SWEEPS. In each group, 3 irrigation/activation cycles of 20 s of irrigation and 20 s of activation were performed with the relevant activation method. The root canals were filled with fluorescein sodium (Sigma, Alldrich Co., St. Louis, MO, USA) and activated with the relevant activation method for 30 s. Specimens were sectioned horizontally to 1 ± 0.1 mm at 2, 5, and 8 mm from the apex and then examined under the CLSM. Maximum penetration depth, maximum penetration area and penetration percentage were measured by using Image J software. Data were analyzed using a two-way analysis of variance (ANOVA) and posthoc Tukey tests (p = .05). No difference was found between the activation systems in terms of maximum penetration depth and maximum penetration area of irrigation solutions in the apical section (p > .05). The penetration percentage of irrigation solutions was higher in PUI and PIPS compared with EA and XPF at the apical section (p < .05). No difference was found between SNI, PUI, PIPS, and SWEEPS in terms of the penetration percentage of irrigation solutions in all sections (p > .05). The penetration percentage of irrigation solutions was higher in the middle and coronal sections compared to the apical section in EA and XPF (p < .05). Tubule penetration of irrigation solutions in SNI was similar between sections. The Tubule penetration area of irrigation solutions in PUI, EA, XPF, PIPS, and SWEEPS was lower in the apical section than in the middle and coronal sections. RESEARCH HIGHLIGHTS: The tubule penetration depth and area of the irrigation solutions were similar between the activation systems tested in the apical sections. The Tubule penetration area of irrigation solutions in middle and coronal sections was higher in PUI and PIPS than in SNI.

Asunto(s)

Hidróxido de Calcio , Dentina , Microscopía Confocal , Irrigantes del Conducto Radicular , Preparación del Conducto Radicular , Irrigación Terapéutica , Microscopía Confocal/métodos , Humanos , Irrigación Terapéutica/métodos , Dentina/efectos de los fármacos , Preparación del Conducto Radicular/métodos , Cavidad Pulpar/efectos de los fármacos , Incisivo

RESUMEN

Mitochondria are the energy factories of a cell, and depending on the metabolic requirements, the mitochondrial morphology, quantity, and membrane potential in a cell change. These changes are frequently assessed using commercially available probes. In this study, we tested the suitability of three commercially available probes-namely 5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolo-carbocyanine iodide (JC-1), MitoTracker Red CMX Rox (CMXRos), and tetramethylrhodamine methyl ester (TMRM)-for assessing the mitochondrial quantity, morphology, and membrane potential in living human mesoangioblasts in 3D with confocal laser scanning microscope (CLSM) and scanning disk confocal microscope (SDCM). Using CLSM, JC-1, and CMXRos-but not TMRM-uncovered considerable background and variation. Using SDCM, the background signal only remained apparent for the JC-1 monomer. Repetitive imaging of CMXRos and JC-1-but not TMRM-demonstrated a 1.5-2-fold variation in signal intensity between cells using CLSM. The use of SDCM drastically reduced this variation. The slope of the relative signal intensity upon repetitive imaging using CLSM was lowest for TMRM (-0.03) and highest for CMXRos (0.16). Upon repetitive imaging using SDCM, the slope varied from 0 (CMXRos) to a maximum of -0.27 (JC-1 C1). Conclusively, our data show that TMRM staining outperformed JC-1 and CMXRos dyes in a (repetitive) 3D analysis of the entire mitochondrial quantity, morphology, and membrane potential in living cells.

Asunto(s)

Imagenología Tridimensional , Microscopía Confocal , Mitocondrias , Humanos , Mitocondrias/metabolismo , Microscopía Confocal/métodos , Imagenología Tridimensional/métodos , Colorantes Fluorescentes/química , Potencial de la Membrana Mitocondrial , Carbocianinas/química , Rodaminas/química

RESUMEN

The label-free imaging of inorganic nanoparticles (NPs) using confocal laser scanning microscopy (CLSM) provides a powerful and versatile tool for studying interactions between NPs and biological systems. Without the need for exogenous labels or markers, it simply benefits from the differential scattering of visible photons between biomaterials and inorganic NPs. Validation experiments conducted on fixed and living cells in real-time, as well as mouse tissue sections following parenteral administration of NPs. Additionally, by incorporating reporter fluorophores and utilizing both reflectance and fluorescence imaging modalities, the method enables high-resolution multiplex imaging of cellular structures and NPs. Different sizes and concentrations of Au NPs are tested as for Ag, Fe3O4, and CeO2 NPs, all with biological interest. Overall, the comprehensive study of NP imaging by confocal microscopy in reflectance mode provides valuable insights and tools for researchers interested in monitoring the nano-bio interactions.

RESUMEN

The stripping reaction of lithium (Li) will greatly impact the cyclability and safety of Li-metal batteries. However, Li pits' nucleation and growth, the origin of uneven stripping, are still poorly understood. In this study, we analyze the nucleation mechanism of Li pits and their morphology evolution with a large population and electrode area (>0.45 cm2). We elucidate the dependence of the pit size and density on the current density and overpotential, which are aligned with classical nucleation theory. With a confocal laser scanning microscope, we reveal the preferential stripping on certain crystal grains and a new stripping mode between pure pitting and stripping without pitting. Descriptors like circularity and the aspect ratio (R) of the pit radius to depth are used to quantify the evolution of Li pits in three dimensions. As the pits grow, growth predominates along the through-planedirection, surpassing the expanding rate in the in-plane direction. After analyzing more than 1000 pits at each condition, we validate that the overpotential is inversely related to the pit radius and exponentially related to the rate of nucleation. With this established nucleation-overpotential relationship, we can better understand and predict the evolution of the surface area and roughness of Li electrodes under different stripping conditions. The knowledge and methodology developed in this work will significantly benefit Li-metal batteries' charging/discharging profile design and the assessment of large-scale Li-metal foils.

RESUMEN

Numerous studies have investigated the effects of stannous ions on specific microbes and their efficacy in reducing dental plaque. Nonetheless, our understanding of their impact on the oral microbiome is still a subject of ongoing exploration. Therefore, this study sought to evaluate the effects of a stannous-containing sodium fluoride dentifrice in comparison to a zinc-containing sodium fluoride dentifrice and a control group on intact, healthy oral biofilms. Utilizing the novel 2bRAD-M approach for species-resolved metagenomics, and FISH/CLSM with probes targeting periodontal and caries associated species alongside Sn2+ and Zn2+ ions, we collected and analyzed in situ biofilms from 15 generally healthy individuals with measurable dental plaque and treated the biofilms with dentifrices to elucidate variations in microbial distribution. Although significant shifts in the microbiome upon treatment were not observed, the use of a stannous-containing sodium fluoride dentifrice primarily led to an increase in health-associated commensal species and decrease in pathogenic species. Notably, FISH/CLSM analysis highlighted a marked reduction in representative species associated with periodontitis and caries following treatment with the use of a stannous-containing sodium fluoride dentifrice, as opposed to a zinc-containing sodium fluoride dentifrice and the control group. Additionally, Sn2+ specific intracellular imaging reflected the colocalization of Sn2+ ions with P. gingivalis but not with other species. In contrast, Zn2+ ions exhibited non-specific binding, thus suggesting that Sn2+ could exhibit selective binding toward pathogenic species. Altogether, our results demonstrate that stannous ions could help to maintain a healthy oral microbiome by preferentially targeting certain pathogenic bacteria to reverse dysbiosis and underscores the importance of the continual usage of such products as a preventive measure for oral diseases and the maintenance of health.

RESUMEN

The antimicrobial activity of Origanum vulgare var. hirtum (O) and Coridothymus capitatus (C) essential oils (EOs) and hydrolates (HYs) of the same botanical species was evaluated on sixteen L. monocytogenes strains from food and clinical origins. The antimicrobial activity was assessed by Minimum Inhibitory Concentration (MIC) determination, viable cell enumeration over time up to 60 min, and evaluation of the cellular damage through Confocal Laser Scanning Microscope (CLSM) analysis. EOs exhibited antimicrobial activity with MIC values ranging from 0.3125 to 10 µL/mL. In contrast, HYs demonstrated antimicrobial effectiveness at higher concentrations (125-500 µL/mL). The effect of HYs was rapid after the contact with the cells, and the cell count reduction over 60 min of HY treatment was about 1.2-1.7 Log CFU/mL. L. monocytogenes cells were stressed by HY treatment, and red cell aggregates were revealed through CLSM observation. Moreover, the combinations of EOs and HYs had an additive antilisterial effect in most cases and allowed the concentration of use to be reduced, while maintaining or improving the antimicrobial effectiveness. The combined use of EOs and HYs can offer novel opportunities for applications, thereby enhancing the antimicrobial effectiveness and diminishing the concentration of use. This provides the added benefit of reducing toxicity and mitigating any undesirable sensory effects.