RESUMO
Tissue-clearing techniques have revolutionized the field of biological imaging by rendering biological specimens transparent and enabling inside optical detection. Light-sheet fluorescence microscopy (LSFM) is a powerful tool for three-dimensional imaging of large biological samples. Combining tissue-clearing techniques with LSFM has advanced the efficient 3D visualization of these samples. A crucial challenge with LSFM is the requirement for the objective to operate within the clearing reagent, which can cause aberrations. To address this issue, we introduce a novel, to our knowledge, approach for the flexible design of the solid immersion refractive meniscus lens (SIMlens), facilitating the use of air objectives with cleared samples. Compared to the previous SIMlens, this method not only eliminates aberrations but also offers customized options for enhancing the numerical aperture and working distance of the objective lens, achieving at least a 10% improvement. We have demonstrated the feasibility of this new method using mouse brain samples.
Assuntos
Lentes , Microscopia de Fluorescência , Animais , Camundongos , Microscopia de Fluorescência/métodos , Encéfalo/diagnóstico por imagem , Imageamento TridimensionalRESUMO
Clarification of the cytotoxic function of T cells is crucial for understanding human immune responses and immunotherapy procedures. Here, we report a high-throughput Bessel oblique plane microscopy (HBOPM) platform capable of 3D live imaging and phenotyping of chimeric antigen receptor (CAR)-modified T-cell cytotoxicity against cancer cells. The HBOPM platform has the following characteristics: an isotropic subcellular resolution of 320 nm, large-scale scouting over 400 interacting cell pairs, long-term observation across 5 hours, and quantitative analysis of the Terabyte-scale 3D, multichannel, time-lapse image datasets. Using this advanced microscopy platform, several key subcellular events in CAR-T cells are captured and comprehensively analyzed; these events include the instantaneous formation of immune synapses and the sustained changes in the microtubing morphology. Furthermore, we identify the actin retrograde flow speed, the actin depletion coefficient, the microtubule polarization and the contact area of the CAR-T/target cell conjugates as essential parameters strongly correlated with CAR-T-cell cytotoxic function. Our approach will be useful for establishing criteria for quantifying T-cell function in individual patients for all T-cell-based immunotherapies.
Assuntos
Imageamento Tridimensional , Imunoterapia Adotiva , Microtúbulos , Receptores de Antígenos Quiméricos , Linfócitos T , Humanos , Receptores de Antígenos Quiméricos/metabolismo , Receptores de Antígenos Quiméricos/imunologia , Imageamento Tridimensional/métodos , Imunoterapia Adotiva/métodos , Linfócitos T/imunologia , Linfócitos T/metabolismo , Microtúbulos/metabolismo , Linhagem Celular Tumoral , Sinapses Imunológicas/imunologia , Sinapses Imunológicas/metabolismo , Citotoxicidade Imunológica , Actinas/metabolismo , Microscopia/métodos , FenótipoRESUMO
Chitosan, as a biomaterial, has increasingly garnered attention. However, its limited solubility in water-only dissolving in certain dilute acidic solutions-substantially restricts its broader application. In this investigation, chitosan underwent a solubilization modification to acquire water solubility, facilitating its dissolution in neutral aqueous mediums. Subsequently, this water-soluble chitosan (WSC) was interlinked with oxidized carboxymethyl cellulose (OCMC), characterized by varied oxidation extents, to synthesize hydrogels. Structural characterization verified the formation of imine bonds resulting from crosslinking interactions between the amino groups of water-soluble chitosan and the aldehyde groups of oxidized carboxymethyl cellulose. Employing performance characterization analysis, it was discerned that an increase in the oxidation level of the oxidized carboxymethyl cellulose corresponded to a denser hydrogel network architecture and the hardness increased from 3.01 N to 6.16 N. Moreover, the capacity of these hydrogels to adsorb methylene blue was meticulously examined. Notably, the hydrogel denoted as WSC/66%OCMC manifested an adsorption capability of 28.08 mg/g for methylene blue. Analytical findings from adsorption kinetics and isotherm studies indicate that the adsorption mechanism of the WSC/66%OCMC hydrogel follows the pseudo-second-order kinetic model and corresponds to the Freundlich isotherm model.
RESUMO
In this study, an innovative ratiometric fluorescence and smartphone-assisted visual sensing platform based on blue-yellow dual-emission carbon dots (BY-CDs) was constructed for the first time to determine brilliant blue. The BY-CDs was synthesized via a facile one-step hydrothermal process involving propyl gallate and o-phenylenediamine. The synthesized BY-CDs exhibit favorable water solubility and exceptional fluorescence stability. Under excitation at 370 nm, BY-CDs show two distinguishable fluorescence emission bands (458 and 558 nm). Upon addition of brilliant blue, the fluorescence intensity at 558 nm exhibited a significant quenching effect attributed to fluorescence resonance energy transfer (FRET), while the fluorescence intensity at 458 nm was basically unchanged. The prepared BY-CDs can effectively serve as a ratiometric nanosensor for determining brilliant blue with the ratio of fluorescence intensities at 458 and 558 nm (F458/F558) as response signal. In addition, the developed ratiometric fluorescence sensor exhibits a noticeable alteration in color from yellow to green under UV light with a wavelength of 365 nm upon addition of varying concentrations of brilliant blue, which provides the possibility of visual detection of brilliant blue by a smartphone application. Finally, the BY-CDs based dual-mode sensing platform successfully detected brilliant blue in actual food samples and achieved a desirable recovery rate. This study highlights the merits of fast, convenient, economical, real-time, visual, high accuracy, excellent precision, good selectivity and high sensitivity for brilliant blue detection, and paves new paths for the monitoring of brilliant blue in real samples.
RESUMO
To understand neural basis of animal behavior, it is necessary to monitor neural activity and behavior in freely moving animal before building relationship between them. Here we use light sheet fluorescence microscope (LSFM) combined with microfluidic chip to simultaneously capture neural activity and body movement in small freely behaving Drosophila larva. We develop a transfer learning based method to simultaneously track the continuously changing body posture and activity of neurons that move together using a sub-region tracking network with a precise landmark estimation network for the inference of target landmark trajectory. Based on the tracking of each labelled neuron, the activity of the neuron indicated by fluorescent intensity is calculated. For each video, annotation of only 20 frames in a video is sufficient to yield human-level accuracy for all other frames. The validity of this method is further confirmed by reproducing the activity pattern of PMSIs (period-positive median segmental interneurons) and larval movement as previously reported. Using this method, we disclosed the correlation between larval movement and left-right asymmetry in activity of a group of unidentified neurons labelled by R52H01-Gal4 and further confirmed the roles of these neurons in bilateral balance of body contraction during larval crawling by genetic inhibition of these neurons. Our method provides a new tool for accurate extraction of neural activities and movement of freely behaving small-size transparent animals.
Assuntos
Larva , Aprendizado de Máquina , Neurônios , Postura , Animais , Larva/fisiologia , Neurônios/fisiologia , Postura/fisiologia , Microscopia de Fluorescência/métodos , Drosophila melanogaster/fisiologia , Drosophila/fisiologia , Movimento/fisiologia , Comportamento Animal/fisiologiaRESUMO
AIM: To investigate the association between central serous chorioretinopathy (CSC) and Helicobacter pylori (Hp) by summarizing all available evidence. METHODS: The Scopus, Embase, EBSCO, PubMed, Web of Science, and Cochrane Library databases for all relevant studies published from inception to October 2022 were searched, and manually searched for relevant reference lists as a supplement. Studies investigating the association between CSC and Hp infection were included. Finally, 8 case-control studies were included in the Meta-analysis after study selection. RESULTS: The results showed no significant correlation between Hp infection and CSC [odds ratio (OR) 1.89, 95% confidential interval (CI) 0.58-6.15, I 2=96%, P=0.29]. After subgroup analysis based on the degree of development of the study (developing/developed countries), it was found that the results of the two subgroups were the same as the whole, and no significant difference between the two subgroups existed. Meta-regression showed that the effect of sample size on heterogeneity among studies was more prominent (P<0.01, adjusted R 2=89.72%), which can explain 89.72% of the sources of heterogeneity. CONCLUSION: This Meta-analysis reveals no significant correlation between Hp infection and CSC, which still warrants further well-designed extensive sample studies to reach a more reliable conclusion and promote a better understanding of the treatment of CSC.
RESUMO
Escherichia coli MLB (MG1655 ΔpflB ΔldhA), which can hardly grow on glucose with little succinate accumulation under anaerobic conditions. Two-stage fermentation is a fermentation in which the first stage is used for cell growth and the second stage is used for product production. The ability of glucose consumption and succinate production of MLB under anaerobic conditions can be improved significantly by using acetate as the solo carbon source under aerobic condition during the two-stage fermentation. Then, the adaptive laboratory evolution (ALE) of growing on acetate was applied here. We assumed that the activities of succinate production related enzymes might be further improved in this study. E. coli MLB46-05 evolved from MLB and it had an improved growth phenotype on acetate. Interestingly, in MLB46-05, the yield and tolerance of succinic acid in the anaerobic condition of two-stage fermentation were improved significantly. According to transcriptome analysis, upregulation of the glyoxylate cycle and the activity of stress regulatory factors are the possible reasons for the elevated yield. And the increased tolerance to acetate made it more tolerant to high concentrations of glucose and succinate. Finally, strain MLB46-05 produced 111 g/L of succinic acid with a product yield of 0.74 g/g glucose. SYNOPSIS.
RESUMO
BACKGROUND: Disinfection byproducts (DBPs) have been shown to impair thyroid function in experimental models. However, epidemiological evidence is scarce. METHODS: This study included 1190 women undergoing assisted reproductive technology (ART) treatment from the Tongji Reproductive and Environmental (TREE) cohort from December 2018 to August 2021. Serum thyrotropin (TSH), free triiodothyronine (FT3), and free thyroxine (FT4) were measured as indicators of thyroid function. FT4/FT3 and TSH/FT4 ratios were calculated as markers of thyroid hormone homeostasis. Dichloroacetic acid (DCAA) and trichloroacetic acid (TCAA), the two most abundant HAAs, in urine were detected to assess individual DBP exposures. RESULTS: After adjusting for relevant covariates, positive associations were observed between urinary TCAA concentrations and serum TSH and TSH/FT4 levels (e.g., percent change = 5.82 %, 95 % CI: 0.70 %, 11.21 % for TSH), whereas inverse associations were found for serum FT3 and FT4 (e.g., percent change = -1.29 %, 95 % CI: -2.49 %, -0.07 % for FT3). There also was a negative association between urinary DCAA concentration and serum FT4/FT3 (percent change = -2.49 %, 95 % CI: -4.71 %, -0.23 %). These associations were further confirmed in the restricted cubic spline and generalized additive models with linear or U-shaped dose-response relationships. CONCLUSION: Urinary HAAs were associated with altered thyroid hormone homeostasis among women undergoing ART treatment.
Assuntos
Glândula Tireoide , Humanos , Feminino , Adulto , Tiroxina/sangue , Tri-Iodotironina/sangue , Tireotropina/sangue , Hormônios Tireóideos/sangue , Testes de Função Tireóidea , Desinfetantes , Acetatos , ChinaRESUMO
This paper presents the formation of a self-healing hydrogel prepared by carboxyethyl modification of chitosan and crosslinking with oxidized sodium alginate. Concurrently, the incorporation of Ca2+ facilitated the formation of "calcium bridges" through intricate coordination with carboxyl moieties, bolstering the attributes of the hydrogel. Various characterization methods, including scanning electron microscopy, texture analysis, and rheological measurements, demonstrated that the introduction of carboxyethyl groups resulted in a more compact hydrogel network structure and improved the hardness and elasticity. The addition of Ca2+ helped to further enhance the mechanical performance of the hydrogel and increase its thermal stability. Then, the adsorption capacity was also investigated, showing adsorption capacities of 46.17 mg/g methylene blue and 46.44 mg/g congo red for carboxyethyl chitosan/oxidized sodium alginate hydrogel, a four-fold increase for congo red versus chitosan/oxidized sodium alginate hydrogel. In addition, the adsorption behavior of CEC/OSA/2%Ca2+ hydrogel can be well described by pseudo-second-order kinetic model and Langmuir adsorption isothermal model. Compared to traditional hydrogels, CEC/OSA/2%Ca2+ hydrogel shows superior mechanical strength, enhanced thermal stability, and improved adsorption capacity, which can effectively adsorb not only methylene blue but also congo red. These advancements demonstrate our hydrogel's innovative properties.
Assuntos
Quitosana , Poluentes Químicos da Água , Quitosana/química , Alginatos/química , Hidrogéis/química , Vermelho Congo , Azul de Metileno/química , Adsorção , Cinética , Poluentes Químicos da Água/química , Concentração de Íons de HidrogênioRESUMO
Solution-processed colloidal quantum dots (CQDs) are promising candidates for broadband photodetectors from visible light to shortwave infrared (SWIR). However, large-size PbS CQDs sensitive to longer SWIR are mainly exposed with nonpolar (100) facets on the surface, which lack robust passivation strategies. Herein, an innovative passivation strategy that employs planar cation, is introduced to enable face-to-face coupling on (100) facets and strengthen halide passivation on (111) facets. The defect density of CQDs film (Eg ≈ 0.74 eV) is reduced from 2.74 × 1015 to 1.04 × 1015 cm-3, coupled with 0.1 eV reduction in the activation energy of defects. The resultant CQDs photodiodes exhibit a low dark current density of 14 nA cm-2 with a high external quantum efficiency (EQE) of 62%, achieving a linear dynamic range of 98 dB, a -3dB bandwidth of 103 kHz and a detectivity of 4.7 × 1011 Jones. The comprehensive performance of the CQDs photodiodes outperforms previously reported CQDs photodiodes operating at >1.6 µm. By monolithically integrated with thin-film transistor (TFT) readout circuit, the broadband CQDs imager covering 0.35-1.8 µm realizes the functions including silicon wafer perspectivity and material discrimination, showing its potential for wide range of applications.
RESUMO
This study successfully grafted caffeic acid and 3,4-dihydroxybenzoic acid into chitosan through a coupling reaction, yielding grafting ratio of 8.93 % for caffeic acid grafted chitosan (CA-GC) and 9.15 % for 3,4-dihydroxybenzoic acid grafted chitosan (DHB-GC) at an optimal concentration of 4 mmol phenolic acids. The characterization of modified chitosans through ultraviolet visible spectrometer (UV-vis), Fourier transform infrared spectrometer (FTIR), proton nuclear magnetic resonance (1H NMR), and x-ray photoelectron spectrometer (XPS) confirmed the successful grafting of phenolic acids. In the subsequent step of emulsion preparation, confocal laser scanning microscope images confirmed the formation of O/W (oil-in-water) emulsions. The phenolic acid-grafted chitosans exhibited better emulsification properties compared to native chitosan, such as reduced droplet size, more uniform emulsion droplet distribution, increased ζ-potential, and enhanced emulsifying activity and stability. Moreover, the modified chitosans demonstrated increased antioxidant activities (evidenced by DPPH and ß-carotene assays) and displayed greater antimicrobial effects against E. coli and S. aureus. Its efficacy in curcumin encapsulation was also notable, with improved encapsulation efficiency, sustained release rates, and enhanced storage and photostability. These findings hint at the potential of modified chitosans as an effective emulsifier.
Assuntos
Quitosana , Curcumina , Hidroxibenzoatos , Quitosana/química , Curcumina/farmacologia , Curcumina/química , Emulsões/química , Escherichia coli , Staphylococcus aureus , Ácidos CafeicosRESUMO
Powdered infant formula (PIF) is prone to Cronobacter sakazakii (C. sakazakii) contamination, which can result in infections that endanger the lives of newborns and infants. Slightly acidic electrolytic water (SAEW) has shown antibacterial effects on a variety of foodborne pathogens and has a wide applicability in the food industry. Here, the antibacterial activity of SAEW against C. sakazakii and its use as a disinfectant on contact surfaces with high infection transmission risk were investigated. The inactivation of SAEW on C. sakazakii was positively correlated to the SAEW concentration and treatment time. The antibacterial effect of SAEW was achieved by decreasing the intracellular adenosine triphosphate (ATP), K+, protein, and DNA contents of C. sakazakii, reducing the intracellular pH (pHin) and destroying the cell morphology, which led to inactivation of C. sakazakii ultimately. To test the applicability of this study, the results showed that approximately 103 CFU/cm2 of C. sakazakii were successfully inactivated on stainless steel and rubber surfaces after a 30 mg/L SAEW treatment for 20 s. These results indicate the antibacterial mechanism and potential application of SAEW against C. sakazakii, as well as a new strategy for the prevention and control of C. sakazakii on stainless steel and rubber surfaces.
RESUMO
BACKGROUND: Vesical Imaging-Reporting and Data System (VI-RADS) is a pathway for the standardized imaging and reporting of bladder cancer staging using multiparametric (mp) MRI. PURPOSE: To investigate additional role of morphological (MOR) measurements to VI-RADS for the detection of muscle-invasive bladder cancer (MIBC) with mpMRI. STUDY TYPE: Retrospective. POPULATION: A total of 198 patients (72 MIBC and 126 NMIBC) underwent bladder mpMRI was included. FIELD STRENGTH/SEQUENCE: 3.0 T/T2-weighted imaging with fast-spin-echo sequence, spin-echo-planar diffusion-weighted imaging and dynamic contrast-enhanced imaging with fast 3D gradient-echo sequence. ASSESSMENT: VI-RADS score and MOR measurement including tumor location, number, stalk, cauliflower-like surface, type of tumor growth, tumor-muscle contact margin (TCM), tumor-longitudinal length (TLL), and tumor cellularity index (TCI) were analyzed by three uroradiologists (3-year, 8-year, and 15-year experience of bladder MRI, respectively) who were blinded to histopathology. STATISTICAL TESTS: Significant MOR measurements associated with MIBC were tested by univariable and multivariable logistic regression (LR) analysis with odds ratio (OR). Area under receiver operating characteristic curve (AUC) with DeLong's test and decision curve analysis (DCA) were used to compared the performance of unadjusted vs. adjusted VI-RADS. A P-value <0.05 was considered statistically significant. RESULTS: TCM (OR 9.98; 95% confidence interval [CI] 4.77-20.8), TCI (OR 5.72; 95% CI 2.37-13.8), and TLL (OR 3.35; 95% CI 1.40-8.03) were independently associated with MIBC at multivariable LR analysis. VI-RADS adjusted by three MORs achieved significantly higher AUC (reader 1 0.908 vs. 0.798; reader 2 0.906 vs. 0.855; reader 3 0.907 vs. 0.831) and better clinical benefits than unadjusted VI-RADS at DCA. Specially in VI-RADS-defined equivocal lesions, MOR-based adjustment resulted in 55.5% (25/45), 70.4% (38/54), and 46.4% (26/56) improvement in accuracy for discriminating MIBC in three readers, respectively. DATA CONCLUSION: MOR measurements improved the performance of VI-RADS in detecting MIBC with mpMRI, especially for equivocal lesions. LEVEL OF EVIDENCE: 3 TECHNICAL EFFICACY: Stage 2.
Assuntos
Imageamento por Ressonância Magnética , Invasividade Neoplásica , Neoplasias da Bexiga Urinária , Humanos , Neoplasias da Bexiga Urinária/diagnóstico por imagem , Neoplasias da Bexiga Urinária/patologia , Masculino , Feminino , Estudos Retrospectivos , Idoso , Pessoa de Meia-Idade , Imageamento por Ressonância Magnética/métodos , Bexiga Urinária/diagnóstico por imagem , Bexiga Urinária/patologia , Estadiamento de Neoplasias , Meios de Contraste , Imageamento por Ressonância Magnética Multiparamétrica/métodos , Idoso de 80 Anos ou mais , Reprodutibilidade dos Testes , Adulto , Curva ROCRESUMO
Prenatal exposures to phthalates and bisphenols have been shown to be linked with adverse birth outcomes. Oxidative stress (OS) is considered a potential mechanism. The objective of this study was to explore the individual and mixtures of prenatal exposures to phthalates and bisphenols in associations with OS biomarkers. We measured eight phthalate metabolites and three bisphenols in the urine samples from 105 pregnant women in Wuhan, China. Urinary 8-hydroxydeoxyguanosine (8-OHdG), 8-isoprostaglandin F2α (8-isoPGF2α), and 4-hydroxy-2-nonenal-mercapturic acid (HNE-MA) were determined as OS biomarkers. The OS biomarkers in associations with the individual chemicals were estimated by linear regression models and restricted cubic spline (RCS) models, and their associations with the chemical mixtures were explored by quantile g-computation (qg-comp) models. In single-pollutant analyses, five phthalate metabolites including monomethyl phthalate (MMP), monoethyl phthalate (MEP), mono-(2-ethylhexyl) phthalate (MEHP), (2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), and mono (2-ethyl-5-oxohexyl) phthalate (MEOHP) were positively associated with urinary 8-OHdG levels (all FDR-adjusted P = 0.06). These associations were further confirmed by the RCS models and were linear (P for overall association ≤ 0.05 and P for non-linear association > 0.05). In mixture analyses, qg-comp models showed that a one-quartile increase in the chemical mixtures of phthalate metabolites and bisphenols was positively associated with urinary levels of 8-OHdG and 8-isoPGF2α, and bisphenol A (BPA) and bisphenol F (BPF) were the most contributing chemicals, respectively. Prenatal exposures to individual phthalates and mixtures of phthalates and bisphenols were associated with higher OS levels.
Assuntos
Compostos Benzidrílicos , Dietilexilftalato/análogos & derivados , Poluentes Ambientais , Fenóis , Ácidos Ftálicos , Efeitos Tardios da Exposição Pré-Natal , Humanos , Feminino , Gravidez , Poluentes Ambientais/análise , Ácidos Ftálicos/metabolismo , Biomarcadores/metabolismo , 8-Hidroxi-2'-Desoxiguanosina , Estresse Oxidativo , Exposição Ambiental/análiseRESUMO
In this paper, the low-velocity impact behavior and damage modes of carbon/glass-hybrid fiber-reinforced magnesium alloy laminates (FMLs-H) and pure carbon-fiber-reinforced magnesium alloy laminates (FMLs-C) are investigated using experimental, theoretical modeling, and numerical simulation methods. Low-velocity impact tests were conducted at incident energies of 20 J, 40 J, and 60 J using a drop-weight impact tester, and the load-displacement curves and energy-time curves of the FMLs were recorded and plotted. The results showed that compared with FMLs-C, the stiffness of FMLs-H was slightly reduced, but the peak load and energy absorption were both greatly improved. Finally, a finite element model based on the Abaqus-VUMAT subroutine was developed to simulate the experimental results, and the damage modes of the metal layer, fiber layer, and interlayer were observed and analyzed. The experimental results are in good agreement with the finite element analysis results. The damage mechanisms of two kinds of FMLs under low-velocity impacts are discussed, providing a reference for the design and application of laminates.
RESUMO
Importance: Numerous studies have documented the association of self-rated health (SRH) with chronic diseases. However, few studies have investigated its association with semen quality. Objective: To examine the association of SRH with semen quality among men undergoing assisted reproductive technology (ART) in China. Design, Setting, and Participants: This cross-sectional study recruited male partners in couples undergoing ART treatment at the Center for Reproductive Medicine, Tongji Hospital, Wuhan, China. A total of 1262 men underwent 2 semen examinations and completed a questionnaire on SRH between December 2018 and January 2020. Data analysis was performed from November 20, 2022, to March 24, 2023. Exposure: SRH, including overall physical and mental health, as well as reproductive-related physical and mental health specifically, were reported at baseline recruitment. Main Outcomes and Measures: Sperm concentration, sperm progressive motility, sperm motility, and sperm count as semen quality parameters. Results: The study included 1262 men with a mean (SD) age of 32.79 (5.25) years and body mass index of 24.37 (3.68). Men with poorer SRH had lower semen quality (eg, sperm concentration among poor vs very good overall physical health: percentage variation, -14.67%; 95% CI, -23.62% to -4.66%). Among 4 components of SRH, a greater reduction in semen quality was estimated for reproductive-related SRH compared with overall SRH, whereas the greatest reduction was observed for reproductive-related physical SRH. In comparison with men with very good reproductive-related physical SRH, men with poor reproductive-related physical SRH had differences of -24.78% (95% CI, -32.71% to -15.93%) and -25.61% (95% CI, -33.95% to -16.22%) in sperm count and concentration, respectively, and regression coefficients of -9.38 (95% CI, -12.01 to -6.76) and -9.24 (95% CI, -11.82 to -6.66) for sperm motility and sperm progressive motility, respectively. Conclusions and Relevance: In this cross-sectional study of Chinese men, poorer SRH was associated with lower semen quality, and reproductive-related physical SRH was the most pronounced indicator. Our findings suggest that SRH, especially reproductive-related physical SRH, was a good indicator of semen quality, which should inform public and clinical regulatory decisions.
Assuntos
Análise do Sêmen , Motilidade dos Espermatozoides , Masculino , Humanos , Adulto , Estudos Transversais , Sêmen , Técnicas de Reprodução AssistidaRESUMO
Rationale: The composition and spatial structure of the lymphoma tumor microenvironment (TME) provide key pathological insights for tumor survival and growth, invasion and metastasis, and resistance to immunotherapy. However, the 3D lymphoma TME has not been well studied owing to the limitations of current imaging techniques. In this work, we take full advantage of a series of new techniques to enable the first 3D TME study in intact lymphoma tissue. Methods: Diverse cell subtypes in lymphoma tissues were tagged using a multiplex immunofluorescence labeling technique. To optically clarify the entire tissue, immunolabeling-enabled three-dimensional imaging of solvent-cleared organs (iDISCO+), clear, unobstructed brain imaging cocktails and computational analysis (CUBIC) and stabilization to harsh conditions via intramolecular epoxide linkages to prevent degradation (SHIELD) were comprehensively compared with the ultimate dimensional imaging of solvent-cleared organs (uDISCO) approach selected for clearing lymphoma tissues. A Bessel-beam light-sheet fluorescence microscope (B-LSFM) was developed to three-dimensionally image the clarified tissues at high speed and high resolution. A customized MATLAB program was used to quantify the number and colocalization of the cell subtypes based on the acquired multichannel 3D images. By combining these cutting-edge methods, we successfully carried out high-efficiency 3D visualization and high-content cellular analyses of the lymphoma TME. Results: Several antibodies, including CD3, CD8, CD20, CD68, CD163, CD14, CD15, FOXP3 and Ki67, were screened for labeling the TME in lymphoma tumors. The 3D imaging results of the TME from three types of lymphoma, reactive lymphocytic hyperplasia (RLN), diffuse large B-cell lymphoma (DLBCL), and angioimmunoblastic T-cell lymphoma (AITL), were quantitatively analyzed, and their cell number, localization, and spatial correlation were comprehensively revealed. Conclusion: We present an advanced imaging-based method for efficient 3D visualization and high-content cellular analysis of the lymphoma TME, rendering it a valuable tool for tumor pathological diagnosis and other clinical research.
Assuntos
Imageamento Tridimensional , Linfoma Difuso de Grandes Células B , Humanos , Imageamento Tridimensional/métodos , Microambiente Tumoral , Microscopia de Fluorescência/métodos , Imunofluorescência , Linfoma Difuso de Grandes Células B/patologia , SolventesRESUMO
Cronobacter sakazakii is an important foodborne pathogen in powder infant formula (PIF). The objective of this study was to evaluate the inactivation effect of Rosa roxburghii Tratt pomace crude extract (RRPCE) on C. sakazakii isolated from PIF and to reveal the mechanism of action. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were used to evaluate the inhibitory activity of RRPCE against C. sakazakii. The inhibitory mechanism was revealed from the perspective of effects of RRPCE on intracellular adenosine 5'-triphosphate (ATP), reactive oxygen species (ROS), membrane potential, protein and nucleic acid leakage, and cell morphology of C. sakazakii. The inactivation effects of RRPCE on C. sakazakii in biofilms on stainless steel, tinplate, glass, silica gel, polyethylene terephthalate, and polystyrene to evaluate its potential as a natural disinfectant. The results showed that the MIC and MBC of RRPCE against C. sakazakii were 7.5 and 15 mg/mL, respectively. After treatments with RRPCE, intracellular ATP content decreased significantly while intracellular ROS level increased significantly (p < 0.05). The cell membrane depolarization, large leakage of proteins and nucleic acids, and severely damaged cell morphology also occurred in C. sakazakii treated with RRPCE. In addition, a 20-minute treatment with 2 MIC (15 mg/mL) of RRPCE could inactivate all C. sakazakii (from 6.10 to 6.40 CFU/mL) in biofilms on all six contact surfaces. Our findings suggest that RRPCE is ideal for the inactivation of C. sakazakii and has the potential to be used as a natural disinfectant for the inactivation of PIF packaging materials and containers.
Assuntos
Cronobacter sakazakii , Cronobacter , Desinfetantes , Rosa , Humanos , Lactente , Fórmulas Infantis , Espécies Reativas de Oxigênio/farmacologia , Trifosfato de Adenosina , Desinfetantes/farmacologia , Microbiologia de AlimentosRESUMO
In this study, chlorogenic acid-chitosan (CA-CS) copolymers were prepared with varying Chitosan (CS): chlorogenic acid (CA)ratios and characterized for their water solubility, antioxidant capacity, and emulsions stability. Results showed that CA-CS samples exhibited up to 90.5 % increase in DPPH scavenging efficiency and 20 % increase in hydroxyl radical scavenging efficiency compared to CS alone. CA-CS copolymers used to stabilize oil in water (O/W) emulsions, which were evaluated for their potential in encapsulating and protecting ß-carotene. Microscopic observations revealed homogeneous spherical droplets in stable emulsions, suggesting effective interfacial structures. The selected CA-CS-stabilized O/W emulsions demonstrated encapsulation efficiencies of 74.8 % and 75.26 % for ß-carotene. The CA-CS stabilized O/W emulsions provided the most effective protection against ß-carotene degradation under UV exposure, retaining over 80 % of ß-carotene content after 12 h of testing. These findings indicate that CA-CS-based O/W emulsions show promise as carriers and protectors for bioactive compounds, due to their improved antioxidant capacity, emulsions stability, and protection against degradation.
Assuntos
Quitosana , Ácido Clorogênico , Ácido Clorogênico/química , Emulsões/química , beta Caroteno/química , Quitosana/química , Antioxidantes/farmacologia , Água/químicaRESUMO
In the paper, Nisin was grafted onto native pectin by the 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC·HCl) method. Structure characterisation showed that the carboxyl group of pectin interacted with the amino group of Nisin and formed an amide bond. The highest grafting ratio of the modified pectin was up to 24.89 %. The emulsifying property of modified pectin, significantly improved, and emulsification performance improved with increasing grafting ratio. Emulsifying activity, emulsion stability, Zeta potential, and droplet morphology data demonstrate a notable enhancement in pectin's emulsifying properties due to Nisin's introduction, with the degree of grafting showing a direct correlation with the improvement observed. Pectin-based emulsion is utilized to load curcumin, enhancing its stability and bioavailability. Research findings highlight that the incorporation of Nisin-modified pectin significantly elevates curcumin encapsulation efficiency, while decelerating its release rate. Moreover, the stability of curcumin loaded in the modified pectin under light exposure, alkaline conditions, and long-term storage is also significantly improved. Ultimately, the bioavailability of curcumin escalates from 0.368 to 0.785.