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Journal of Photochemistry and... Jun 2024Bovine mastitis (BM) represents a significant challenge in the dairy industry. Limitations of conventional treatments have prompted the exploration of alternative...
Bovine mastitis (BM) represents a significant challenge in the dairy industry. Limitations of conventional treatments have prompted the exploration of alternative approaches, such as photodynamic inactivation (PDI). In this study, we developed a PDI protocol to eliminate BM-associated pathogens using porphyrin-doped conjugated polymer nanoparticles (CPN). The PDI-CPN protocol was evaluated in four mastitis isolates of Staphylococcus and in a hyper-biofilm-forming reference strain. The results in planktonic cultures demonstrated that PDI-CPN exhibited a bactericidal profile upon relatively low light doses (∼9.6 J/cm). Furthermore, following a seven-hour incubation period, no evidence of cellular reactivation was observed, indicating a highly efficient post-photodynamic inactivation effect. The successful elimination of bacterial suspensions encouraged us to test the PDI-CPN protocol on mature biofilms. Treatment using moderate light dose (∼64.8 J/cm) reduced biofilm biomass and metabolic activity by up to 74% and 88%, respectively. The impact of PDI-CPN therapy on biofilms was investigated using scanning electron microscopy (SEM), which revealed nearly complete removal of the extracellular matrix and cocci. Moreover, ex vivo studies conducted on bovine udder skin demonstrated the efficacy of the therapy in eliminating bacteria from these scaffolds and its potential as a prophylactic method. Notably, the histological analysis of skin revealed no signs of cellular degeneration, suggesting that the protocol is safe and effective for BM treatment. Overall, this study demonstrates the potential of PDI-CPN in treating and preventing BM pathogens. It also provides insights into the effects of PDI-CPN on bacterial growth, metabolism, and survival over extended periods, aiding the development of effective control strategies and the optimization of future treatments.
PubMed: 38955081
DOI: 10.1016/j.jphotobiol.2024.112971 -
International Journal of Surgery... Jul 2024Carbon nanoparticle suspension injection (CNSI) and indocyanine green (ICG) have both been applied intraoperatively to facilitate lymphatic mapping and postoperatively...
Effect of carbon nanoparticle suspension injection versus indocyanine green tracer in guiding lymph node dissection during radical gastrectomy (FUTURE-01): a randomized clinical trial.
BACKGROUND
Carbon nanoparticle suspension injection (CNSI) and indocyanine green (ICG) have both been applied intraoperatively to facilitate lymphatic mapping and postoperatively to sort lymph nodes (LNs) in gastric cancer patients. However, no study has compared the two tracers in gastric cancer patients.
MATERIALS AND METHODS
This prospective randomized controlled trial was conducted from January 2022 to March 2023. Patients with potentially resectable gastric cancer (cT1-4a N0/+ M0) were randomized to the CNSI or ICG group.
RESULTS
This study enrolled 96 patients. Ninety patients were in the modified intention-to-treat population, including 46 patients (32 males and 14 females; mean [SD] age, 57.4 [9.4] years) in the CNSI group and 44 patients (31 males and 13 females; mean [SD] age, 60.8 [8.8] years) in the ICG group. The mean (SD) number of retrieved LNs was 69.8 (21.9) and 53.6 (17.2) in the CNSI and ICG groups, respectively (P<0.001). The mean (SD) number of retrieved micro-LNs was 19.9 (13.3) and 11.6 (9.9) in the CNSI and ICG groups, respectively (P=0.001). The mean (SD) number of metastatic LNs was 8.1 (11.9) and 5.2 (9.2) in the CNSI and ICG groups, respectively (P=0.19).
CONCLUSIONS
Compared with ICG, CNSI can increase the number of LNs detected, especially micro-LNs. Both tracers have high diagnostic value for detecting metastatic LNs. CNSI-guided lymphography may be a superior method for improving the accuracy of LN dissection.
PubMed: 38954670
DOI: 10.1097/JS9.0000000000001873 -
Analytical Chemistry Jul 2024Glycans on proteins and lipids play important roles in maturation and cellular interactions, contributing to a variety of biological processes. Aberrant glycosylation...
Glycans on proteins and lipids play important roles in maturation and cellular interactions, contributing to a variety of biological processes. Aberrant glycosylation has been associated with various human diseases including cancer; however, elucidating the distribution and heterogeneity of glycans in complex tissue samples remains a major challenge. Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) is routinely used to analyze the spatial distribution of a variety of molecules including N-glycans directly from tissue surfaces. Sialic acids are nine carbon acidic sugars that often exist as the terminal sugars of glycans and are inherently difficult to analyze using MALDI-MSI due to their instability prone to in- and postsource decay. Here, we report on a rapid and robust method for stabilizing sialic acid on N-glycans in FFPE tissue sections. The established method derivatizes and identifies the spatial distribution of α2,3- and α2,6-linked sialic acids through complete methylamidation using methylamine and PyAOP ((7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate). Our in situ approach increases the glycans detected and enhances the coverage of sialylated species. Using this streamlined, sensitive, and robust workflow, we rapidly characterize and spatially localize N-glycans in human tumor tissue sections. Additionally, we demonstrate this method's applicability in imaging mammalian cell suspensions directly on slides, achieving cellular resolution with minimal sample processing and cell numbers. This workflow reveals the cellular locations of distinct N-glycan species, shedding light on the biological and clinical significance of these biomolecules in human diseases.
PubMed: 38953530
DOI: 10.1021/acs.analchem.3c05984 -
Cureus Jun 2024Background MXene is a newly discovered substance consisting of 2D transition metal carbides or nitrides, produced through the disintegration and etching of aluminum...
Background MXene is a newly discovered substance consisting of 2D transition metal carbides or nitrides, produced through the disintegration and etching of aluminum layers. It possesses numerous properties, including a high surface area, conductivity, strength, stiffness, negative zeta potential, and excellent volumetric capacitance. MXene is utilized in detecting anti-cancer medicine, while bismuth vanadate (BiVO) is synthesized to form an optimized material for anti-cancer activity applications. BiVO exhibits visible light absorption, strong chemical stability, and non-toxic properties. However, when loaded onto target stem cells, it can cause skin and respiratory irritation. Aim This study aimed to evaluate the facile fabrication of titanium carbide (TiC)-BiVO nanomaterials coupled with oxides for anti-cancer activity. Moreover, it aimed to create TiC-BiVO nanomaterials in combination with oxides using X-ray diffraction (XRD) and scanning electron microscopy (SEM) to assess their potential as efficient and targeted anti-cancer agents. Methods and materials To prepare the 2D TiC MXene, 2.5 g of titanium aluminum carbide (TiAlC) powder was dissolved in 60 mL of a 40% hydrofluoric acid (HF) solution in a polytetrafluoroethylene(PTFE) container. The etching process was made more efficient and completed in 24 hours by using a magnetic stirring system to keep the mixture stirred and heated continuously. The centrifugation was performed at 4000 rpm for five minutes. Subsequently, deionized water was used to wash the solution many times until its pH reached around 7. The appropriate TiC powder was made by vacuum drying the acquired sediment at 80°C for 24 hours. Monoclinic BiVO samples were synthesized via a hydrothermal method. Typically, 10 mmol of Bi(NO).5HO was dissolved in 100 mL of a 2 mol/L HNO solution and stirred uniformly. Subsequently, 10 mmol of ammonium metavanadate (NHVO) was added to the mixed solution. After being stirred for one hour, the mixture was transferred into a 100 mL sealed Teflon-lined stainless steel autoclave at 180°C for 16 hours. After cooling to room temperature, the sediment was washed three times with deionized water, ethanol, and acetone, respectively. Finally, the suspension was dried at 80°C, followed by calcination at 450°C for three hours to obtain BiVO. TiC-BiVO heterostructures were prepared by surface modification TiCusing BiVO suspensions by a simple, cost-effective approach. Results TiC nanosheets were observed with BiVO particles, and the high crystalline nature of the compound was confirmed after XRD analysis and energy-dispersive spectroscopy (EDS) analysis. The compound was found to be pure without any impurities and exhibited anti-cancer activity. Conclusion The XRD, field emission scanning electron microscopy(FESEM), and EDS investigations provide an in-depth analysis of the structural, morphological, and compositional characteristics of TiC-BiVO sheets. The XRD analysis proves the successful combination of different materials and the presence of crystalline phases. The FESEM imaging technique exposes the shape and arrangement of particles in sheets, while the EDS analysis verifies the elemental composition and uniform distribution. These investigations show that TiC-BiVO composites have been successfully synthesized, indicating their potential for use in anti-cancer applications.
PubMed: 38952587
DOI: 10.7759/cureus.61492 -
Soft Matter Jul 2024We present a simple route to obtain large quantities of suspensions of non-Brownian particles with -responsive surface properties to study the relation between their...
We present a simple route to obtain large quantities of suspensions of non-Brownian particles with -responsive surface properties to study the relation between their flow and interparticle interactions. We perform an alkaline hydrolysis reaction on poly(methyl methacrylate) (PMMA) particles to obtain poly(sodium methacrylate) (PMAA-Na) particles. We characterize the quasi-static macroscopic frictional response of their aqueous suspensions using a rotating drum. The suspensions are frictionless when the particles are dispersed in pure water. We relate this state to the presence of electrosteric repulsion between the charged surfaces of the ionized PMAA-Na particles in water. Then we add monovalent and multivalent ions (Na, Ca, La) and we observe that the suspensions become frictional whatever the valency. For divalent and trivalent ions, the quasi-static avalanche angle at large ionic strength is greater than that of frictional PMMA particles in water, suggesting the presence of adhesion. Finally, a decrease in the pH of the suspending solution leads to a transition between a frictionless plateau and a frictional one. We perform atomic force microscopy (AFM) to relate our macroscopic observations to the surface features of the particles. In particular, we show that the increase in friction in the presence of multivalent ions or under acidic conditions is driven by a nanoscopic phase separation and the bundling of polyelectrolyte chains at the surface of the particle. Our results highlight the importance of surface interactions in the rheology of granular suspensions. Our particles provide a simple, yet flexible platform to study frictional suspension flows.
PubMed: 38952147
DOI: 10.1039/d4sm00381k -
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi =... Jun 2024Objective To investigate the effects of astragaloside IV(AS-IV) on the balance of T helper type 1 (Th1) and Th2 cells in mice with IgA nephropathy (IgAN) and its...
Objective To investigate the effects of astragaloside IV(AS-IV) on the balance of T helper type 1 (Th1) and Th2 cells in mice with IgA nephropathy (IgAN) and its possible mechanism. Methods The IgAN model of BALB/c mice was established. Successfully modeled mice were randomly divided into four groups: model, AS-IV low dose, AS-IV medium dose and AS-IV high dose groups, with 10 mice in each group. Another 10 mice served as the control group. Mice in the low, medium and high dose groups were administered 12.5, 25 and 50 mg/kg AS-IV suspension (prepared in normal saline) by gavage, while the control and model groups were given an equivalent volume of normal saline. The 24-hour urinary protein (24 h UPr) content and urine red blood cell count were measured in each group. The levels of blood urea nitrogen (BUN), serum creatinine (Scr) and albumin (ALB) were determined. Serum interferon γ (IFN-γ), interleukin 4 (IL-4) and IL-10 levels were detected by ELISA. The ratio of Th1/Th2 cells in peripheral blood of mice was detected using flow cytometry. Histopathological changes in the kidney of mice were observed by HE staining. RT-PCR and Western blot were used to detect the mRNA and protein expressions of T cell immunoglobulin and mucin domain gene 1 (TIM-1), Toll-like receptor 4 (TLR4) in mouse kidney tissue. Results Compared with the model group, in weeks 12 and 15, the urine red blood cell count, 24 h UPr, BUN, Scr, levels of IL-4 and IL-10, the proportion of Th2 cells, as well as the mRNA and protein expression levels of TIM-1 and TLR4 were significantly decreased in the low, medium and high dose groups of AS-IV, and the levels of ALB, IFN-γ, the proportion of Th1 cells and Th1/Th2 cell ratio were increased, with the high-dose group showing the best effects. Conclusion AS-IV can inhibit TIM-1 signaling pathway, increase the Th1/Th2 cell ratio, inhibit the inflammatory reaction, and alleviate the renal injury in IgAN mice.
Topics: Animals; Hepatitis A Virus Cellular Receptor 1; Triterpenes; Glomerulonephritis, IGA; Saponins; Th1 Cells; Signal Transduction; Mice, Inbred BALB C; Th2 Cells; Mice; Toll-Like Receptor 4; Interleukin-4; Kidney; Interleukin-10; Interferon-gamma; Male; Female
PubMed: 38952089
DOI: No ID Found -
Methods in Molecular Biology (Clifton,... 2024Mammalian cell lines are one of the best options when it comes to the production of complex proteins requiring specific glycosylation patterns. Plasmid DNA transfection...
Mammalian cell lines are one of the best options when it comes to the production of complex proteins requiring specific glycosylation patterns. Plasmid DNA transfection and stable cell lines are frequently used for recombinant protein production, but they are expensive at large scale or can become time-consuming, respectively. The BacMam baculovirus (BV) is a safe and cost-effective platform to produce recombinant proteins in mammalian cells. The process of generating BacMam BVs is straightforward and similar to the generation of "insect" BVs, with different commercially available platforms. Although there are several protocols that describe recombinant protein expression with the BacMam BV in adherent cell lines, limited information is available on suspension cells. Therefore, it is of relevance to define the conditions to produce recombinant proteins in suspension cell cultures with BacMam BVs that facilitate bioprocess transfer to larger volumes. Here, we describe a method to generate a high titer BacMam BV stock and produce recombinant proteins in suspension HEK293 cells.
Topics: Baculoviridae; Humans; Recombinant Proteins; HEK293 Cells; Animals; Transfection; Genetic Vectors; Cell Culture Techniques; Gene Expression; Glycosylation
PubMed: 38951347
DOI: 10.1007/978-1-0716-3961-0_25 -
Methods in Molecular Biology (Clifton,... 2024Nonviral transfection has been used to express various recombinant proteins, therapeutics, and virus-like particles (VLP) in mammalian and insect cells. Virus-free...
Nonviral transfection has been used to express various recombinant proteins, therapeutics, and virus-like particles (VLP) in mammalian and insect cells. Virus-free methods for protein expression require fewer steps for obtaining protein expression by eliminating virus amplification and measuring the infectivity of the virus. The nonviral method uses a nonlytic plasmid to transfect the gene of interest into the insect cells instead of using baculovirus, a lytic system. In this chapter, we describe one of the transfection methods, which uses polyethyleneimine (PEI) as a DNA delivery material into the insect cells to express the recombinant protein in both adherent and suspension cells.
Topics: Animals; Recombinant Proteins; Transfection; Polyethyleneimine; Plasmids; Insecta; Sf9 Cells; Cell Line; Gene Expression; Spodoptera
PubMed: 38951345
DOI: 10.1007/978-1-0716-3961-0_23 -
Methods in Molecular Biology (Clifton,... 2024This chapter outlines the workflow using the ExpiSf Expression System designed for high-density infection of suspension ExpiSf9 cells. The system utilizes a chemically...
This chapter outlines the workflow using the ExpiSf Expression System designed for high-density infection of suspension ExpiSf9 cells. The system utilizes a chemically defined, serum-free, protein-free, and animal origin free medium, making it suitable for recombinant protein expression experiments. The ExpiSf chemically defined medium allows efficient transfection and baculovirus production directly within the same culture medium. The ExpiSf Expression System Starter Kit provides all necessary components, including cells, culture medium, and reagents needed to infect one (1) liter of cell culture. The system's versatility and animal origin free nature make it a valuable tool for various protein expression studies and biotechnological applications.
Topics: Animals; Workflow; Recombinant Proteins; Baculoviridae; Transfection; Culture Media; Cell Culture Techniques; Cell Line; Gene Expression
PubMed: 38951326
DOI: 10.1007/978-1-0716-3961-0_4 -
ChemPlusChem Jul 2024The adsorption characteristics of novel activated biocarbons prepared from horsetail herb by physical activation (using carbon dioxide) and chemical one (using...
The adsorption characteristics of novel activated biocarbons prepared from horsetail herb by physical activation (using carbon dioxide) and chemical one (using phosphoric(V) acid) in the process of simultaneous proteins immobilization in multicomponent solutions were examined. The carbon materials were characterized in terms of their porous structure, acidic-basic properties, and surface morphology. The binding mechanisms of such proteins as bovine serum albumin (BSA) and lysozyme (LSZ), differing in internal stability, were determined alone and in their blends. This was done based on the comprehensive analysis of the results of adsorption/desorption, surface, electrokinetic and stability measurements. These experiments were carried out over a wide pH range of 3-11. They included the following issues: (1) determination of the protein adsorbed/desorbed amounts on/from a surface of activated biocarbons; (2) study of the kinetics of these processes; (3) examination of the macromolecules impact on the surface charge density and zeta potential of the carbon materials; and (4) determination of the suspension stability and size of aggregates formed in the examined systems. The analysis of the obtained results indicated the differences in the binding mechanism of both proteins that is of key importance for their simultaneous immobilization on activated biocarbons surface in the soil environment.
PubMed: 38951113
DOI: 10.1002/cplu.202400177