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Regenerative Therapy Jun 2024With the increasing emphasis on the use of nonanimal ingredients in clinical care, studies have proposed the use of TrypLE™ as an alternative to trypsin. However,...
INTRODUCTION
With the increasing emphasis on the use of nonanimal ingredients in clinical care, studies have proposed the use of TrypLE™ as an alternative to trypsin. However, previous research has reported insufficient cell yield and viability when using TrypLE to isolate skin cells compared to the dispase/trypsin-EDTA method. This study aimed to propose an improved method for increasing the yield and viability of cells isolated by TrypLE and to evaluate isolated keratinocytes and melanocytes.
METHODS
Foreskin tissues were isolated to keratinocytes and melanocytes using the trypsin-EDTA protocol and our modified TrypLE protocol. The yield and viability of freshly isolated cells were compared, the epidermal residue after cell suspension filtration was analyzed histologically, and the expression of cytokeratin 14 (CK14) and Melan-A was detected by flow cytometry. After cultivation, keratinocytes and melanocytes were further examined for marker expression and proliferation. A coculture model of melanocytes and HaCaT cells was used to evaluate melanin transfer.
RESULTS
The yield, viability of total cells and expression of the keratinocyte marker CK14 were similar for freshly isolated cells from both protocols. No differences were observed in the histologic analysis of epidermal residues. Moreover, no differences in keratinocyte marker expression or melanocyte melanin transfer function were observed after culture. However, melanocytes generated using the TrypLE protocol exhibited increased Melan-A expression and proliferation in culture.
CONCLUSION
Our TrypLE protocol not only solved the problems of insufficient cell yield and viability in previous studies but also preserved normal cell morphology and function, which enables the clinical treatment of depigmentation diseases.
PubMed: 38948130
DOI: 10.1016/j.reth.2024.05.014 -
ACS Omega Jun 2024This study conducts a systematic investigation of the creation and optimization of a rutin-loaded transethosome intended for topical use. The formulation's...
This study conducts a systematic investigation of the creation and optimization of a rutin-loaded transethosome intended for topical use. The formulation's characteristics were thoroughly assessed for vesicle size (160.45 ± 1.98 nm), polydispersity index (0.235 ± 0.067), and zeta potential (-22.89 mV), with an entrapment efficiency and drug loading of 89.99 ± 1.55% and 8.9 ± 2.11%, respectively, and found to have a spherical shape by the use of transmission electron microscopy. The conversion to a gel suitable for application on the skin was carried out. The drug release form Opt-RUT-TE formulation (73.61 ± 2.55%) was significantly higher than that of release form RUT-suspension (34.52 ± 1.19%). The drug that permeated the skin from Opt-RUT-TEG (935.25 ± 10.49 μg/cm) was significantly higher than the permeability from RUT-Suspension gel (522.57 ± 6.79 μg/cm). Notably, tape stripping analysis revealed that the Opt-RUT-TE gel effectively penetrated the skin layers, with a higher concentration observed in the epidermis-dermis than in the RUT-suspension gel. The transethosomal gel exhibited favorable characteristics, highlighting its capacity to efficiently permeate the skin and suppress the growth of microorganisms, and Opt-RUT-TEG showed a higher microorganism inhibition zone (Gram-positive bacteria) than that of RUT-suspension gel. The investigation highlights the significant therapeutic possibilities of rutin in a transethosomal gel formulation for treating dermatological diseases by improving skin permeability and exhibiting antibacterial effects.
PubMed: 38947795
DOI: 10.1021/acsomega.4c01718 -
Neuropsychiatric Disease and Treatment 2024Absence seizures are classically associated with behavioral arrest and transient deficits in consciousness, yet substantial variability exists in the severity of the... (Review)
Review
Absence seizures are classically associated with behavioral arrest and transient deficits in consciousness, yet substantial variability exists in the severity of the impairment. Despite several decades of research on the topic, the pathophysiology of absence seizures and the mechanisms underlying behavioral impairment remain unclear. Several rationales have been proposed including widespread cortical deactivation, reduced perception of external stimuli, and transient suspension of the default mode network, among others. This review aims to summarize the current knowledge on the neural correlates of impaired consciousness in absence seizures. We review evidence from studies using animal models of absence epilepsy, electroencephalography, functional magnetic resonance imaging, magnetoencephalography, positron emission tomography, and single photon emission computed tomography.
PubMed: 38947367
DOI: 10.2147/NDT.S391052 -
Research Square Jun 2024Organoid cultures offer a powerful technology to investigate many different aspects of development, physiology, and pathology of diverse tissues. Unlike standard tissue...
Organoid cultures offer a powerful technology to investigate many different aspects of development, physiology, and pathology of diverse tissues. Unlike standard tissue culture of primary breast epithelial cells, breast organoids preserve the epithelial lineages and architecture of the normal tissue. However, existing organoid culture methods are tedious, difficult to scale, and do not robustly retain estrogen receptor (ER) expression and responsiveness in long-term culture. Here, we describe a modified culture method to generate and maintain organoids as suspension cultures in reconstituted basement membrane (™Matrigel). This method improves organoid growth and uniformity compared to the conventional Matrigel dome embedding method, while maintaining the fidelity of the three major epithelial lineages. Using this adopted method, we are able to culture and passage purified hormone sensing (HS) cells that retain ER responsiveness upon estrogen stimulation in long-term culture. This culture system presents a valuable platform to study the events involved in initiation and evolution of ER-positive breast cancer.
PubMed: 38947074
DOI: 10.21203/rs.3.rs-4463390/v1 -
Analytical Methods : Advancing Methods... Jul 2024High-frequency pulse lasers, applied in the form of rapid scanning, act upon the surface of aircraft skin paint layers, thereby removing the paint layers, exhibiting...
High-frequency pulse lasers, applied in the form of rapid scanning, act upon the surface of aircraft skin paint layers, thereby removing the paint layers, exhibiting characteristics of efficiency and eco-friendliness. Real-time monitoring of the paint removal effect and prevention of substrate damage necessitates the continuous monitoring of paint removal thickness. Combining Laser-Induced Breakdown Spectroscopy (LIBS) online monitoring technology enables laser-controlled paint removal under multiple effects coupling, meeting the requirements of airworthiness maintenance. This paper, based on a high-frequency nanosecond infrared pulse laser paint removal LIBS monitoring platform, conducts research on laser paint removal thickness LIBS online monitoring of aluminum alloy plates coated with dual-layer paint. Spectra corresponding to the removal thickness of each group are collected and, respectively, paint removal thickness monitoring models based on LIBS spectra are established using the standard curve method and Principal Component Analysis-Support Vector Regression (PCA-SVR) algorithm. When monitoring paint removal thickness using the standard curve method, the intensity of five Ti element characteristic spectral lines selected is correlated with the paint removal thickness, and segmented curve fitting according to the paint layer structure satisfies the segmented curve fitting of topcoat and topcoat + primer. Among them, the average coefficient of the curve fitting of the Ti II 589.088 nm characteristic spectral line is 0.89, and the root mean square error (RMSE) is 12.28 μm. Its performance is superior in the five standard curves; thus, its fitting equation is used as the criterion for paint removal thickness monitoring. To further improve monitoring accuracy, research on paint removal thickness monitoring models based on PCA-SVR is conducted. Compared to the traditional univariate standard curve method, the PCA-SVR model does not require segmented monitoring. After parameter optimization, the average fitting coefficient reaches 0.97, and the RMSE is 2.92 μm. The results indicate that the PCA-SVR-based paint removal thickness monitoring model has higher accuracy, thereby forming the basis for paint removal thickness monitoring. Through comparative research on paint removal thickness monitoring models, two types of paint removal thickness monitoring criteria are obtained, providing model solutions for high-precision monitoring and automation of aircraft skin laser paint removal thickness.
PubMed: 38946640
DOI: 10.1039/d4ay00872c -
Journal of Materials Chemistry. B Jul 2024Micro- and nanoencapsulation of cells has been studied as a strategy to protect cells from environmental stress and promote survival during delivery. Hydrogels used in...
Micro- and nanoencapsulation of cells has been studied as a strategy to protect cells from environmental stress and promote survival during delivery. Hydrogels used in encapsulation can be modified to influence cell behaviors and direct assembly in their surroundings. Here, we report a system that conformally encapsulated stem cells using hyaluronic acid (HA). We successfully modified HA with lipid, thiol, and maleimide pendant groups to facilitate a hydrogel system in which HA was deposited onto cell plasma membranes and subsequently crosslinked through thiol-maleimide click chemistry. We demonstrated conformal encapsulation of both neural stem cells (NSCs) and mesenchymal stromal cells (MSCs), with viability of both cell types greater than 90% after encapsulation. Additional material could be added to the conformal hydrogel through alternating addition of thiol-modified and maleimide-modified HA in a layering process. After encapsulation, we tracked egress and viability of the cells over days and observed differential responses of cell types to conformal hydrogels both according to cell type and the amount of material deposited on the cell surfaces. Through the design of the conformal hydrogels, we showed that multicellular assembly could be created in suspension and that encapsulated cells could be immobilized on surfaces. In conjunction with photolithography, conformal hydrogels enabled rapid assembly of encapsulated cells on hydrogel substrates with resolution at the scale of 100 μm.
PubMed: 38946474
DOI: 10.1039/d4tb00223g -
Magnetic Resonance in Medicine Jun 2024Serine is a major source of one-carbon units needed for the synthesis of nucleotides and the production of intramitochondrial nicotinamide adenine dinucleotide phosphate...
PURPOSE
Serine is a major source of one-carbon units needed for the synthesis of nucleotides and the production of intramitochondrial nicotinamide adenine dinucleotide phosphate (NADPH), and it plays an important role in cancer cell proliferation. The aim of this study was to develop a deuterium (H) MRS imaging method for imaging tumor serine metabolism.
METHODS
Sequential (H) spectra and spectroscopic images were used to monitor the metabolism of [2,3,3-H]serine in patient-derived glioblastoma cells in vitro and in tumors obtained by their orthotopic implantation in mouse brain.
RESULTS
[14,14-H] 5,10-methylene-tetrahydrofolate, [H]glycine, [H]formate, and labeled water were detected in cell suspensions and water labeling in spectroscopic images of tumors. Studies in cells and tumors with variable mitochondrial content and inhibitor studies in cells demonstrated that most of the labeled serine was metabolized in the mitochondria. Water labeling in the cell suspensions was correlated with formate labeling; therefore, water labeling observed in tumors could be used to provide a surrogate measure of flux in the pathway of one-carbon metabolism in vivo.
CONCLUSION
The method has the potential to be used clinically to select patients for treatment with inhibitors of one-carbon metabolism and subsequently to detect their early responses to such treatment.
PubMed: 38946234
DOI: 10.1002/mrm.30198 -
Food Research International (Ottawa,... Aug 2024Fungal contaminations of cereal grains are a profound food-safety and food-security concern worldwide, threatening consumers' and animals' health and causing enormous...
Fungal contaminations of cereal grains are a profound food-safety and food-security concern worldwide, threatening consumers' and animals' health and causing enormous economic burdens. Because far-ultraviolet C (far-UVC) light at 222 nm has recently been shown to be human-safe, we investigated its efficacy as an alternative to thermal, chemical, and conventional 254 nm UVC anti-fungal treatments. Our microplasma-based far-UVC lamp system achieved a 5.21-log reduction in the conidia of Aspergillus flavus suspended in buffer with a dose of 1032.0 mJ/cm, and a 5.11-log reduction of Fusarium graminearum conidia in suspension with a dose of 619.2 mJ/cm. We further observed that far-UVC treatments could induce fungal-cell apoptosis, alter mitochondrial membrane potential, lead to the accumulation of intracellular reactive oxygen species, cause lipid peroxidation, and result in cell-membrane damage. The lamp system also exhibited a potent ability to inhibit the mycelial growth of both A. flavus and F. graminearum. On potato dextrose agar plates, such growth was completely inhibited after doses of 576.0 mJ/cm and 460.8 mJ/cm, respectively. To test our approach's efficacy at decontaminating actual cereal grains, we designed a cubical 3D treatment chamber fitted with six lamps. At a dose of 780.0 mJ/cm on each side, the chamber achieved a 1.88-log reduction of A. flavus on dried yellow corn kernels and a 1.11-log reduction of F. graminearum on wheat grains, without significant moisture loss to either cereal type (p > 0.05). The treatment did not cause significant changes in the propensity of wheat grains to germinate in the week following treatment (p > 0.05). However, it increased the germination propensity of corn kernels by more than 71% in the same timeframe (p < 0.05). Collectively, our results demonstrate that 222 nm far-UVC radiation can effectively inactivate fungal growth in liquid, on solid surfaces, and on cereal grains. If scalable, its emergence as a safe, cost-effective alternative tool for reducing fungi-related post-harvest cereal losses could have important positive implications for the fight against world hunger and food insecurity.
Topics: Fusarium; Aspergillus flavus; Edible Grain; Ultraviolet Rays; Spores, Fungal; Food Contamination; Food Irradiation; Food Microbiology; Reactive Oxygen Species
PubMed: 38945594
DOI: 10.1016/j.foodres.2024.114550 -
Brain Research Jun 2024Prolonged confinement in cramped spaces can lead to derangements in brain function/structure, yet the underlying mechanisms remain unclear. To investigate, we subjected...
Prolonged confinement in cramped spaces can lead to derangements in brain function/structure, yet the underlying mechanisms remain unclear. To investigate, we subjected mice to restraint stress to simulate long-term narrow and enclosed space confinement, assessing their mental state through behavioral tests. Stressed mice showed reduced center travel and dwell time in the Open Field Test and increased immobility in the Tail Suspension Test. We measured lower hippocampal brain-derived neurotrophic factor levels and cortical monoamine neurotransmitters (5-HT and NE) in the stressed group. Further examination of the body's immune levels and serum metabolism revealed immune dysregulation and metabolic imbalance in the stressed group. The results of the metabolic network regulation analysis indicate that the targets affected by these differential metabolites are involved in several metabolic pathways that the metabolites themselves participate in, such as the "long-term depression" and "purine metabolism" pathways. Additionally, these targets are also associated with numerous immune-related pathways, such as the TNF, NF-κB, and IL-17 signaling pathways, and these findings were validated using GEO dataset analysis. Molecular docking results suggest that differential metabolites may regulate specific immune factors such as TNF-α, IL-1β, and IL-6, and these results were confirmed in experiments. Our research findings suggest that long-term exposure to confined and narrow spaces can lead to the development of psychopathologies, possibly mediated by immune system dysregulation and metabolic disruption.
PubMed: 38945470
DOI: 10.1016/j.brainres.2024.149101 -
Journal of Plant Physiology Jun 2024Programmed cell death (PCD) is a genetically regulated process of cell suicide essential for plant development. The 'malate valve' is a mechanism that ensures redox...
Programmed cell death (PCD) is a genetically regulated process of cell suicide essential for plant development. The 'malate valve' is a mechanism that ensures redox balance across different subcellular compartments. In broccoli, the BomMDH1 gene encodes malate dehydrogenase in mitochondria, a critical enzyme in the 'malate circulation' pathway. This study investigates the functional role of BomMDH1 in malate (MA)-induced apoptosis in bright yellow-2 (BY-2) suspension cells. Findings revealed that transgenic cells overexpressing BomMDH1 showed enhanced viability under MA-induced oxidative stress compared to wild-type (WT) cells. Overexpression of BomMDH1 also reduced levels of reactive oxygen species (ROS), hydrogen peroxide (HO), and malondialdehyde (MDA), while increasing the expression of antioxidant enzyme genes such as NtAPX, NtAOX1a, NtSOD, and NtMDHAR. Additionally, treatment with salicylhydroxamic acid (SHAM), a characteristic inhibitor of mitochondrial respiration, further improved the anti-apoptotic activity of BY-2 cells. Overall, these results highlighted the function of the BomMDH1 gene and the potential of SHAM treatment in mitigating oxidative stress in BY-2 suspension cells.
PubMed: 38945071
DOI: 10.1016/j.jplph.2024.154297