Confocal Laser Tomography Research Articles

Adaptation mechanism of clam Ruditapes philippinarum under polycyclic aromatic hydrocarbon stresses: Accumulation and excretion

Persistent organic pollutants (POPs) are easily accumulated in organisms due to its high lipophilicity. Bivalves are still one of the marine invertebrates with high species diversity despite experiencing with environmental pollution. However, studies investigating the adaptation mechanisms of bivalves towards POPs have been lacking. In this study, we chose benzo[a]pyrene (B[a]P), a typical POPs, to investigate the accumulation and excretion mechanism of clam Ruditapes philippinarum towards B[a]P. Laser confocal microscopy images of ovaries and testes confirmed significant colocalization between lipid droplets and B[a]P. However, B[a]P in the testes might mainly excrete with the excretory fluid. Furthermore, lipidomics results indicated that the upregulated TGs were TGs with 48–58 total carbon atoms and 6–12 double bonds in acyl side chains in ovaries, while the upregulated TGs mainly distributed in 48–58 total carbon atoms and 4–14 double bonds in acyl side chains in testes. In ovaries, gene expression of lipid metabolism and phospholipid metabolism basically presented an upward trend at Pro and Mat stages, while entirely performed a downregulation trend at Gro stage. Testes had the opposite regulation with that. Additionally, B[a]P prompted the excretion ability of female clams, but exhibited a complex effect in the male. This study provides a scientific basis for bivalve toxicology and healthy aquaculture, which is of great significance for marine ecological environment protection and aquatic product safety of China.

The influence of pore throat heterogeneity and fractal characteristics on reservoir quality: A case study of chang 8 member tight sandstones, Ordos Basin

The Chang 8 member of the Yanchang Formation in the Ordos Basin exhibits extensive development of tight sandstone reservoirs. In comparison to conventional reservoirs, these tight sandstone reservoirs demonstrate smaller pore-throat systems and stronger heterogeneity due to interactions between inherent sedimentary components and various diagenetic processes. To further investigate the influence of microscopic pore throat structure on macroscopic physical properties and oil content within the reservoir, a comprehensive study was conducted utilizing high pressure mercury injection, nuclear magnetic resonance, X-CT scanning, and other pore throat testing methods. This study was complemented by observation techniques such as polarizing microscope, scanning electron microscopy, and laser confocal imaging, to study the pore throat system of reservoirs under the influence of different sedimentary components and diagenesis. By comparison, it is observed that the radius of the primary pore throat system in the tight sandstone reservoir ranges from 0.01 to 1 μm. The structural characteristics of the pore throat system significantly impact the macroscopic physical properties of the reservoir. Due to variations in testing methods, high pressure mercury injection, nuclear magnetic resonance (NMR), and X-ray computed tomography (X-CT) reveal different fractal dimensions of the reservoir's pore throat system. Therefore, it is better to respectively utilize fractal dimension Df2, Df2-NMR, Df2-CT, and pore throat parameters to characterize the microscopic pore throat system with a radius ranging from 0.01 to 1 μm. The decrease in the average pore throat radius and the increased irregularity of pore shape contribute to heightened heterogeneity in the micro-pore throat structure within the reservoir. This negatively impacts its physical properties and oil content. Furthermore, it is important to note that the minimum threshold of pore throat radius plays a crucial role as a primary factor determining the fluid seepage capacity within this reservoir. Moreover, when isolated pores exist, reduced fluid mobility ensues, which further worsens the seepage conditions of the reservoir. The chlorite rims development reservoir shows a low fractal dimension in its micro-pore throats, contributing to superior macroscopic petrophysical properties and oil content. However, compression, siliceous cementation, and calcite cementation negatively impact the average pore throat radius by increasing micropores, inaccessible pores, and narrow throat content. This results in a stronger heterogeneity of the pore throat structure, leading to poor seepage conditions and lower oil content.

Procyanidin B1 Promotes PSMC3-NRF2 Ubiquitination to Induce Ferroptosis in Glioblastoma.

NRF2 signaling is a crucial antioxidant defense mechanism against ferroptosis in tumors, and targeting NRF2 is essential for tumor therapy. However, the effectiveness of NRF2 inhibitors remains unexplored. The active ingredients of traditional Chinese medicine serve as important sources of NRF2 inhibitors. In this study, we established an intracranial glioblastoma (GBM) orthotopic model and observed the effects of procyanidin B1 on tumor growth and ferroptosis. Using protein-small-molecule docking, z-stack assay of laser confocal imaging, surface plasmon resonance assay, immunoprecipitation, mass spectrometry, and western blotting, we detected the binding between procyanidin B1 and NRF2 and the effect of PSMC3 on the ubiquitin-dependent degradation of NRF2 in GBM cells. Our results showed that procyanidin B1 acted as a novel NRF2 inhibitor to suppress GBM cell proliferation and prolonged the survival of GBM-bearing mice; it also mediated the interaction between PSMC3 and NRF2 to promote ubiquitin-dependent protein degradation of NRF2, which induced ferroptosis in GBM cells. In addition, we found that procyanidin B1 enhanced H₂O₂ accumulation by downregulating NRF2 during ferroptosis in GBM cells. The botanical agent procyanidin B1 induced ferroptosis and exerted anti-tumor effects through PSMC3-mediated ubiquitin-dependent degradation of NRF2 proteins, providing a potential drug candidate for adjuvant therapy in patients with GBM.

Epinecidin-1 and lactic acid synergistically inhibit Aeromonas hydrophila through membrane disruption

Epinecidin-1 (Epi-1) is an antimicrobial peptide originated from fish with various pharmacological activities but carries the risk of acquiring resistance with long-term use. In the present study, we use L-lactic acid to enhance the antibacterial activity of synthesized Epi-1 against the aquaculture and food pathogen Aeromonas hydrophila. The results showed that 5.5 mmol/L lactic acid increased the inhibitory and bactericidal activity of 25 μmol/L Epi-1 against two strains of A. hydrophila. The laser confocal images proved that lactic acid pre-treatment improved the attachment efficiency of Epi-1 in A.hydrophila cells. In addition, lactic acid enhanced the damaging effect of Epi-1 on the cell membrane of A. hydrophila, evidenced by releasing more nucleic acids, proteins, and transmembrane pH ingredients decrease and electromotive force dissipation. SEM images showed that compared with the single Epi-1 treatment, the co-treatment of Epi-1 and lactic acid caused more outer membrane vesicles (OMVs) and more severe cell deformation. These findings proved that lactic acid could enhance the efficiency of Epi-1 against A. hydrophila and shed light on new aspects to avoid resistance of pathogens against Epi-1.

The Biological Crystals in Chamid Bivalve Shells: Diversity in Morphology and Crystal Arrangement Pattern

Chamid bivalves are marine organisms that live in high-energy environments and are cemented to hard substrates. To avoid shell damage, the organisms form thick, densely ornamented shells. Shell material consists of aragonite, and the ornamentation may be either aragonitic or calcitic. The latter can be developed as scaly spines, rows of blades, or comarginal, radial arched lamellae. We investigated biological crystal morphology and mode of assembly of Chama arcana and Chama gryphoides shells. Structural characteristics were obtained from electron backscatter diffraction (EBSD) measurements, complemented with laser confocal and BSE imaging. We found a wide range of crystal morphologies and sizes, ranging from irregularly shaped calcite and/or aragonite prisms to tiny and thin aragonite laths. We observed four different modes of crystal assembly patterns: 1. strongly interlocked dendritic calcite units forming the ornamentation blades; 2. aragonite laths arranged to lamellae forming the outer shell layer, the layer adjacent to the calcite; 3. aragonite laths arranged into blocks comprising inner shell layers or aragonitic ornamentations; and 4. shell portions consisting of aragonite prisms, structured in size and crystal orientation, at muscle attachment sites. These four different types of crystal arrangements were observed for the shells of the investigated chamid species; however, they had slightly different strengths of structuring and slight variations in crystal organisation. Additionally, we observed unique microstructural features in Chama shells: We report ornamentation crystals resembling idiomorphic calcite and novel, twinned entities found at the changeover between the aragonitic layers. We highlight and discuss these differences and anomalies in this contribution.

Innovative insights into the preparation of tamarind gum base film loaded with pomelo essential oil via a low-energy emulsion method combined with melatonin: Maximizing the dual functions of tamarind gum

Pomelo essential oil has good antioxidant and antimicrobial properties. The objective of this study was to modify the active packaging loaded with essential oil through low-energy emulsification, in order to achieve both film-forming and emulsifying properties of tamarind gum, and to investigate the surface activity of melatonin in the emulsion. When α:β mixing ratio of 60:40, the resulting film solution exhibited favorable apparent viscosity and reduced emulsion flocculation and precipitation phenomena. Particle size distribution analysis and optical microscopy results demonstrated that the obtained emulsion was uniformly dispersed at an α:β mixing ratio of 60:40, with droplet diameters mainly concentrated within the range of 1–100 μm. After undergoing film-forming treatment, E-3-F (α:β mixing ratio of 60:40) exhibited a smoother and more compact microstructure, leading to significant improvements in mechanical properties and barrier performance. Laser confocal microscopy imaging revealed that the addition of melatonin further reduced interfacial tension between oil and water, while simultaneously weakening hydrophobic interaction forces among oil droplets, thereby effectively inhibiting droplet aggregation behavior. Moreover, incorporation of melatonin also enhanced material compactness and imparted excellent antioxidant effects to the packaging material. Consequently, this study ultimately unveils potential for low-energy emulsification combined with melatonin application in bio-based packaging field while providing novel insights into developing multifunctional bio-based packaging.

Agar-gelatin Maillard conjugates used for Pickering emulsion stabilization

A few protein- and polysaccharide-based particles have shown promising potential as stabilizers in multi-phase food systems. By incorporating polymer-based particles and modifying the wettability of colloidal systems, it is possible to create particle-stabilized emulsions with excellent stability. A Pickering emulsifier (AGMs) with better emulsifying properties was obtained by the Maillard reaction between acid-hydrolysed agar and gelatin. Laser confocal microscopy imaging revealed that AGMs particles can be used as solid emulsifiers to produce a typical O/W Pickering emulsion, with AGMs adsorbing onto the droplet surface to form a dense interfacial layer. Cryo-scanning electron microscopy analysis showed that AGMs self-assembled into a three-dimensional network structure, which prevented droplets aggregation through strong spatial site resistance, contributing to emulsion stabilization. These emulsions exhibited stability within a pH range of 1 to 11, NaCl concentrations not exceeding 300 mM, and at temperatures below 80 °C. The most stable emulsion oil-water ratio was 6:4 at a particle concentration of 0.75 % (w/v). AGMs-stabilized Pickering emulsion was utilized to create a semi-solid mayonnaise as a replacement for hydrogenated oil. Rheological analysis demonstrated that low-fat mayonnaise stabilized with AGMs exhibited similar rheological behavior to traditional mayonnaise, offering new avenues for the application of Pickering emulsions in the food industry.

Effects of soaking glutinous sorghum grains on physicochemical properties of starch

Glutinous sorghum grains were soaked (60–80 °C, 2–8 h) to explore the effects of soaking, an essential step in industrial processing of brewing, on starch. As the soaking temperature increased, the peak viscosity and crystallinity of starch gradually decreased, while the enzymatic hydrolysis rate and storage modulus first increased and then decreased. At 70 °C, the content of amylose, the enzymatic hydrolysis rate of starch, and the final viscosity first increase and then decrease with the increase of soaking time, reaching their maximum at 6 h, increased by 53.1 %, 11.0 %, and 10.4 %, respectively, as compared with the non-soaked sample. At 80 °C (4 h), the laser confocal microscopy images showed a network structure formed between the denatured protein chains and the leached-out amylose chains. The molecular weights of starch before and after soaking were all in the range of 3.82–8.98 × 107 g/mol. Since 70 °C is lower than that of starch gelatinization and protein denaturation, when soaking for 6 h, the enzymatic hydrolysis rate of starch is the highest, and the growth of miscellaneous bacteria is inhibited, which is beneficial for subsequent processing technology. The result provides a theoretical basis for the intelligent control of glutinous sorghum brewing.

Hydrophobic Surface Array Structure Based on Laser-Induced Graphene for Deicing and Anti-Icing Applications.

The exceptional performance of graphene has driven the advancement of its preparation techniques and applications. Laser-induced graphene (LIG), as a novel graphene preparation technique, has been applied in various fields. Graphene periodic structures created by the LIG technique exhibit superhydrophobic characteristics and can be used for deicing and anti-icing applications, which are significantly influenced by the laser parameters. The laser surface treatment process was simulated by a finite element software analysis (COMSOL Multiphysics) to optimize the scanning parameter range, and the linear array surface structure was subsequently fabricated by the LIG technique. The generation of graphene was confirmed by Raman spectroscopy and energy-dispersive X-ray spectroscopy. The periodic linear array structure was observed by scanning electron microscopy (SEM) and confocal laser imaging (CLSM). In addition, CLSM testings, contact angle measurements, and delayed icing experiments were systematically performed to investigate the effect of scanning speed on surface hydrophobicity. The results show that high-quality and uniform graphene can be achieved using the laser scanning speed of 125 mm/s. The periodic linear array structures can obviously increase the contact angle and suppress delayed icing. Furthermore, these structures have the enhanced ability of the electric heating deicing, which can reach 100 °C and 240 °C within 15 s and within 60 s under the DC voltage power supply ranging from 3 to 7 V, respectively. These results indicate that the LIG technique can be developed to provide an efficient, economical, and convenient approach for preparing graphene and that the hydrophobic surface array structure based on LIG has considerable potential for deicing and anti-icing applications.

Machine learning-assisted pattern recognition and imaging of multiplexed cancer cells via a porphyrin-embedded dendrimer array.

Early cancer detection plays a vital role in improving the survival rate of cancer patients, underscoring the importance of developing cancer detection methods. However, it is a great challenge to achieve simple, rapid, and accurate methods for simultaneously discerning various cancers. Herein we developed a 5-element porphyrin-embedded dendrimer-based sensor array, targeting the parallel discrimination of multiple cancers. The porphyrin-embedded dendrimers were modified with various functional groups to generate differentiated interactions with diverse cancer cells, which has been validated by fluorescence responses and laser confocal microscopy imaging. The dual-channel, five-element array, featuring ten signal outputs, achieved 100% accuracy in distinguishing between one human normal cell and six human cancerous cells, as well as in differentiating among mixed cells. Moreover, the screen 6-channel array can accurately distinguish 9 cells from mice and humans in minutes through optimization by multiple machine learning algorithms, including two normal cells and 7 cancerous cells with only 1000 cells, highlighting the significant potential of a porphyrin-embedded dendrimer-based parallel discriminating platform in early cancer diagnosis.

A plasma-3D print combined in vitro platform with implications for reliable materiobiological screening.

Materiobiology is an emerging field focused on the physiochemical properties of biomaterials concerning biological outcomes which includes but is not limited to the biological responses and bioactivity of surface-modified biomaterials. Herein, we report a novel in vitro characterization platform for characterizing nanoparticle surface-modified 3D printed PLA scaffolds. We have introduced innovative design parameters that were practical for ubiquitous in vitro assays like those utilizing 96 and 24-well plates. Subsequently, gold and silica nanoparticles were deposited using two low-temperature plasma-assisted processes namely plasma electroless reduction (PER) and dusty plasma on 3D scaffolds. Materiobiological testing began with nanoparticle surface modification optimization on 96 well plate design 3D scaffolds. We have employed 3D laser confocal imaging and scanning electron microscopy to study the deposition of nanoparticles. It was found that the formation and distribution of the nanoparticles were time-dependent. In vitro assays were performed utilizing an osteosarcoma (MG-63) cell as a model. These cells were grown on both 96 and 24 well plate design 3D scaffolds. Subsequently, we performed different in vitro assays such as cell viability, and fluorescence staining of cytoskeletal actin and DNA incorporation. The actin cytoskeleton staining showed more homogeneity in the cell monolayer growing on the gold nanoparticle-modified 3D scaffolds than the control 3D PLA scaffold. Furthermore, the mineralization and protein adsorption experiments conducted on 96 well plate design scaffolds have shown enhanced mineralization and bovine serum albumin adsorption for the gold nanoparticle-modified scaffolds compared to the control scaffolds. Taken together, this study reports the efficacy of this new in vitro platform in conducting more reliable and efficient materiobiology studies. It is also worth mentioning that this platform has significant futuristic potential for developing as a high throughput screening platform. Such platforms could have a significant impact on the systematic study of biocompatibility and bioactive mechanisms of nanoparticle-modified 3D-printed scaffolds for tissue engineering. It would also provide unique ways to investigate mechanisms of biological responses and subsequent bioactive mechanisms for implantable biomaterials. Moreover, this platform can derive more consistent and reliable in vitro results which can improve the success rate of further in vivo experiments.

Stomatin promotes neutrophil degranulation and vascular leakage in the early stage after severe burn via enhancement of the intracellular binding of neutrophil primary granules to F-actin

BackgroundThe pathophysiology of severe burn injuries in the early stages involves complex emergency responses, inflammatory reactions, immune system activation, and a significant increase in vascular permeability. Neutrophils, crucial innate immune cells, undergo rapid mobilization and intricate pathophysiological changes during this period. However, the dynamic alterations and detailed mechanisms governing their biological behavior remain unclear. Stomatin protein, an essential component of the cell membrane, stabilizes and regulates the membrane and participates in cell signal transduction. Additionally, it exhibits elevated expression in various inflammatory diseases. While Stomatin expression has been observed in the cell and granule membranes of neutrophils, its potential involvement in post-activation functional regulation requires further investigation. MethodsNeutrophils were isolated from human peripheral blood, mouse peripheral blood, and mouse bone marrow using the magnetic bead separation method. Flow cytometry was used to assess neutrophil membrane surface markers, ROS levels, and phagocytic activity. The expression of the Stomatin gene and protein was examined using quantitative real-time polymerase chain reaction and western blotting methods, respectively. Furthermore, the enzyme-linked immunosorbent assay was used to evaluate the expression of neutrophil-derived inflammatory mediators (myeloperoxidase (MPO), neutrophil elastase (NE), and matrix metalloproteinase 9 (MMP9)) in the plasma. Images and videos of vascular leakage in mice were captured using in vivo laser confocal imaging technology, whereas in vitro confocal microscopy was used to study the localization and levels of the cytoskeleton, CD63, and Stomatin protein in neutrophils. ResultsThis study made the following key findings: (1) Early after severe burn, neutrophil dysfunction is present in the peripheral blood characterized by significant bone marrow mobilization, excessive degranulation, and impaired release and chemotaxis of inflammatory mediators (MPO, NE, and MMP9). (2) After burn injury, expression of both the stomatin gene and protein in neutrophils was upregulated. (3) Knockout (KO) of the stomatin gene in mice partially inhibited neutrophil excessive degranulation, potentially achieved via reduced production of primary granules and weakened binding of primary granules to the cell skeleton protein F-actin. (4) In severely burned mice, injury led to notable early-stage vascular leakage and lung damage, whereas Stomatin gene KO significantly ameliorated lung injury and vascular leakage. ConclusionsStomatin promotes neutrophil degranulation in the early stage of severe burn injury via increasing the production of primary granules and enhancing their binding to the cell skeleton protein F-actin in neutrophils. Consequently, this excessive degranulation results in aggravated vascular leakage and lung injury.

Noninvasive imaging of rat-derived microglia and its reactivity to inflammatory molecules via acoustic impedance microscopy.

Microglia, the brain's immune cells, play important roles in neuronal differentiation, survival, and death. The function of microglia is deeply related to the morphologies; however, it is too complex to observe conventionally and identify the condition of living microglia using optical microscopes. Herein, we proposed a new method to observe living cultured microglia and their reactivity to inflammation via the acoustic impedance mode of a scanning acoustic microscope. Primary cultured microglia collected from rat pups exposed to acetamiprid, an insecticide, in utero were observed with both acoustic interface impedance mode (C-mode) and transparent three-dimensional impedance mode (B-mode). We characterized microglia into four types based on the results obtained from acoustic impedance, cytoskeletal information, and laser confocal imaging. Biphasic acoustic observation using B-mode and C-mode gave us information regarding the dynamic morphologies of living microglia treated with adenosine triphosphate (ATP) (600μmol/L), which reflects distress signals from inflamed neurons. Acetamiprid exposure induced microglia response even in the neonatal period. ATP stimulus altered the shape and thickness of microglia with a change in the bulk modulus of the cell. Three-dimensional alteration with ATP stimulus could be observed only after biphasic acoustic observation using B-mode and C-mode. This acoustic observation was consistent with confocal observation using anti-Iba-1 and P2Y12 immunocytochemistry. This study demonstrated the adequacy of using a scanning acoustic microscope in analyzing microglia's shape, motility, and response to inflammation.

Experimental and numerical investigation of damage and mechanical property retention by bio-composite plastic made with flax or pinewood fiber and aged by exposure to ultraviolet light

The objective of this study was to compare two types of natural short fibers, one lignin-rich and one lignin-poor, as reinforcing filler of bio-composite plastic in accelerated aging experiments. Short natural-fiber-reinforced plastic composites of flax fiber or pinewood fiber (30 % by weight) were injected and then exposed to ultraviolet (UV) light with and without moisture for up to 1440 h. Color, roughness, and mechanical properties were then evaluated using colorimetry, confocal laser imaging, tensile tests, and drop-weight tests. Damage to the bio-composites was monitored using the acoustic emission technique during tensile testing. The response during impact tests was simulated using the finite element method. Results showed that flax-fiber-reinforced polypropylene exhibited superior properties before aging, but experienced greater loss of these properties after aging, particularly under moist conditions. The numerical model was found to be in good agreement with the experimental results.

LRRC25 Inhibits IFN-γ Secretion by Microglia to Negatively Regulate Anti-Tuberculosis Immunity in Mice.

Leucine-rich repeat-containing protein-25 (LRRC25) can degrade the ISG15 gene in virus-infected cells and prevent overactivation of the type Ⅰ IFN pathway. However, the role of LRRC25 in bacterial infection is still unclear. In this pursuit, the present study aimed to explore the regulatory role and mechanism of LRRC25 in microglia infected with Mycobacterium tuberculosis in a mouse model. Q-PCR, WB, and cell immunofluorescence were employed to observe the change in LRRC25 in BV2 cells infected by H37Rv. Additionally, siRNA was designed to target the LRRC25 to inhibit its expression in BV2 cells. Flow cytometry and laser confocal imaging were used to observe the infection of BV2 cells after LRRC25 silencing. Q-PCR and ELISA were used to determine the changes in IFN-γ and ISG15 in the culture supernatant of each group. Following H37Rv infection, it was observed that the expression of LRRC25 was upregulated. Upon silencing LRRC25, the proportion of BV2 cells infected by H37Rv decreased significantly. ELISA analysis showed that IFN-γ and ISG15 levels in cell culture supernatant decreased after H37Rv infection, while they significantly increased after LRRC25 silencing. This study provides evidence that LRRC25 is the key negative regulator of microglial anti-Mtb immunity. It exerts its function by degrading free ISG15 and inhibiting the secretion of IFN-γ, thereby improving the anti-Mtb immunity of BV2 cells.

A coumarin based ratiometric fluorescent probe for the detection of Cu2+ and mechanochromism as well as application in living cells and vegetables

In this paper, a novel coumarin-derived fluorescent probe NY was designed and synthesized. NY displayed a significant ratiometric fluorescence response towards Cu2+ in PBS buffer (10 mM, pH = 7.4), with the emission wavelength blue-shifted from 580 to 495 nm, and a fluorescence change from orange to green was evident under a 365 nm UV light. Meanwhile, NY had the advantages of high selectivity, short response time (5 min), low detection limit (1.3 × 10-8 M) and large binding constant (1.45 × 105 M-1) towards Cu2+. The binding mechanism between NY and Cu2+ was elucidated by FT-IR, 1H NMR titration, TOF-MS and Job's plot analysis. In addition, NY was successfully employed in the detection of Cu2+ within environmental water and vegetable samples with satisfactory results. Laser confocal microscopy imaging results showed that NY could easily penetrate HeLa cells membrane to target mitochondria and image Cu2+ in living cells. Furthermore, NY demonstrated mechanochromic properties by exhibiting orange-red fluorescence when subjected to mechanical grinding.

Microdosimetric analysis of monoenergetic electrons and beta-emitting radionuclides based on mesh-type cell models and RBE prediction

AimsMicrodosimetric indicators are commonly used to describe the energy deposition characteristics of ionizing radiation at the subcellular and DNA levels to predict radiation-induced biological effects. Monoenergetic electrons and beta-emitting radionuclides (β nuclides) have a certain range in low-density media, and their microdosimetric indicators in the cell nucleus are of great significance for understanding the characteristics of different types of β nuclides and the evaluation of relative biological effectiveness (RBE). MethodsMicrodosimetry in the nucleus is greatly affected by cell morphology, so in this paper, based on laser confocal tomography images of BEAS-2B and L-O2 cell lines, two types of mesh-type cell models were constructed to restore the morphological features of real cells. The Geant4-DNA toolkit was used to simulate the lineal energy and dose distribution of monoenergetic electrons (0.1 keV-1 MeV) and 5 types of beta nuclides (Cs-137, Cs-134, C-14, I-131, and H-3) in the cell nucleus with different initial energies. The effects of factors such as initial energy, sensitive site (radius = 2 nm, 10 nm, 30 nm, 300 nm), and cell type on microdosimetric quantities in the nucleus and their predictions of nuclide RBE were analysed. ResultsThe results show that for the same type of nuclide, there are different RBE values between BEAS-2B and L-O2 cells, with Cs-134 having a difference of 17.75% and H-3 having a difference of 10.58%. In addition, the mean lineal energy in the nucleus fluctuates with changes in the initial energy and sampling site radius of monoenergetic electrons, especially for low and medium energies where the sampling site radius has a significant effect. Moreover, the proximity functions of monoenergetic electrons with an initial energy below 1 keV are greatly affected by the cell shape, and those above 20 keV are greatly affected by the model volume. ConclusionsThe study confirms the necessity of using mesh cell models for microdosimetric analysis in the cell nucleus and predicting its biological effects by combining mesh models of real human cells to simulate microdosimetric indicators of monoenergetic electrons and beta-emitting nuclides. The estimation of the RBE of β nuclides achieved in this study has reference value for predicting radiobiological damage in different cell types.

Absorption characteristics and the effect on vascular endothelial cell permeability of an anticoagulant peptide

In the former report, the casein peptide TKLTEEEKNR (PfCN) exhibits strong thrombin inhibitory activity in vitro. Its absorption capabilities, however, are unclear. Therefore, we studied its absorption characteristics both in vivo and in vitro. PfCN was carried by cells from the apical chamber to the basolateral chamber via active translocation in Caco-2 cells. Meanwhile, it can also be transported by HUVECs. We found that PfCN can be taken up by HUVECs using confocal laser imaging. PfCN has been proven to have good absorption properties in in vivo experiments. After five minutes of oral treatment, PfCN was identified in the blood, peaking at 82.75 ± 36.52 ng/mL in 30 min. And PfCN vanished from the blood circulation after 120 min. According to in vivo experiments, excessive concentrations of PfCN will alter the permeability of HUVECs. As a result, there is a foundation for PfCN application in the food sector. Meanwhile, we also hope this article can give an idea to the researchers who studying the absorption of functional peptides.

A highly sensitive fluorescent HClO probe for discrimination between cancerous and normal cells/tissues

Elevated level of hypochlorous acid (HClO) is closely associated with cancer development. Identifying HClO level in cancer cells would provide important evidence in either early-stage cancer diagnostics or monitoring of its treatment efficiency. In this work, a new pyronine-based fluorescent probe for rapid and sensitive detection of HClO was developed by condensing meso‑formyl pyronine (PyCHO) with 2-hydrazinopyridine to form meso‑pyridylhydrazone-functionalized pyronine PyHP, PyHP is nonfluorescent due to the excited-state C=N isomerization nonradiative decay, whereas the HClO-triggered formation of meso‑triazolopyridyl pyronine PyTP abolishes the C=N isomerization and thus greatly enhances the fluorescence. With the probe, the cancer cells/tumor were distinguished with high-contrast from normal ones by laser confocal fluorescence imaging, and the tumor-to-normal (T/N) ratios obtained exceed the clinically acceptable threshold of 2.0. Moreover, its capability of in vivo imaging tumor was also demonstrated. These results indicate the potential of PyHP as an effective tool in the early clinical diagnosis of cancers.

Multiple Mesh-type Real Human Cell Models for Dosimetric Application Coupled with Monte Carlo Simulations.

The mesh-type models are superior to voxel models in cellular dose assessment coupled with Monte Carlo codes. The aim of this study was to expand the micron-scale mesh-type models based on the fluorescence tomography of real human cells, and to investigate the feasibility of these models in the application of various irradiation scenarios and Monte Carlo codes. Six different human cell lines, including pulmonary epithelial BEAS-2B, embryonic kidney 293T, hepatocyte L-02, B-lymphoblastoid HMy2.CIR, Gastric mucosal GES-1, and intestine epithelial FHs74Int, were adopted for single mesh-type models reconstruction and optimization based on laser confocal tomography images. Mesh-type models were transformed into the format of polygon mesh and tetrahedral mesh for the GATE and PHITS Monte Carlo codes, respectively. The effect of model reduction was analyzed by dose assessment and geometry consideration. The cytoplasm and nucleus doses were obtained by designating monoenergetic electrons and protons as external irradiation, and S values with different "target-source" combinations were calculated by assigning radioisotopes as internal exposure. Four kinds of Monte Carlo codes were employed, i.e., GATE with "Livermore," "Standard" and "Standard and Geant4-DNA mixed" models for electrons and protons, as well as PHITS with "EGS" mode for electrons and radioisotopes. Multiple mesh-type real human cellular models can be applied to Monte Carlo codes directly without voxelization when combined with certain necessary surface reduction. Relative deviations between different cell types were observed among various irradiation scenarios. The relative deviation of nucleus S value reaches up to 85.65% between L-02 and GES-1 cells by 3H for the "nucleus-nucleus" combination, while that of 293T and FHs74Int nucleus dose for external beams at a 5.12 cm depth of water is 106.99%. Nucleus with smaller volume is far more affected by physical codes. There is a considerable deviation for dose within BEAS-2B at the nanoscale. The multiple mesh-type real cell models were more versatile than voxel models and mathematical models. The present study provided several models which can easily be extended to other cell types and irradiation scenarios for RBE estimations and biological effect predictions, including radiation biological experiments, radiotherapy and radiation protection.