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Search Results (1,052)

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13 pages, 1021 KiB  
Article
Changes in the Intracellular Composition of Macro and Microminerals After Cryopreservation of the Rabbit Stem/Progenitor Cells
by Jaromír Vašíček, Andrej Baláži, Mária Tirpáková, Marián Tomka and Peter Chrenek
J. Dev. Biol. 2025, 13(1), 6; https://doi.org/10.3390/jdb13010006 - 21 Feb 2025
Viewed by 169
Abstract
Cryopreservation is a widely used method for the long-term preservation of reproductive or somatic cells. It is known that this storage method may negatively affect cell viability, proliferation, differentiation, etc. However, there is a lack of information about whether cryostorage can alter the [...] Read more.
Cryopreservation is a widely used method for the long-term preservation of reproductive or somatic cells. It is known that this storage method may negatively affect cell viability, proliferation, differentiation, etc. However, there is a lack of information about whether cryostorage can alter the content of intracellular minerals. Therefore, we focused this study on the analysis of the mineral composition of living cells before and after long-term cold storage. Briefly, three different primary cell lines were established from rabbits as follows: endothelial progenitor cells from peripheral blood (EPCs), endothelial progenitor cells from bone marrow (BEPCs), and mesenchymal stem cells from adipose tissue (AT-MSCs), which were cultured until passage 3 prior to cryopreservation in liquid nitrogen. Samples from freshly cultured and frozen–thawed cells were mineralized and analyzed using inductively coupled plasma-optical emission spectroscopy (ICP-OES) for the content of minerals (macro: Ca, Na, K, and Mg, and micro: Zn, Fe, Cu, Al, Co, Mn, Sr, and Ni). After cryopreservation, we found significantly decreased content of K in frozen–thawed EPCs (p < 0.01) and BEPCs (p < 0.0001) and Ca in AT-MSCs (p < 0.05), while Na was increased in frozen–thawed BEPCs (p < 0.05). Concentrations of Fe and Al were reduced significantly in frozen–thawed EPCs (both p < 0.0001) and AT-MSCs (p < 0.001 and p < 0.0001, respectively). On the contrary, Fe and Al were elevated in frozen–thawed BEPCs (p < 0.0001 and p < 0.01, respectively) together with Ni (p < 0.0001). In addition, decreased Zn (p < 0.05) was observed in cryopreserved AT-MSCs. In conclusion, the ICP-OES technique might be used to analyze the basic elemental composition of animal cells in fresh or frozen–thawed conditions. Nevertheless, additional studies are needed to reveal the possible impact of cryopreservation on cell fate by changing the content of intracellular minerals. Full article
13 pages, 3969 KiB  
Review
Gas Plasma Technology and Immunogenic Cell Death: Implications for Chordoma Treatment
by Sander Bekeschus, Karl Roessler, Oliver Kepp and Eric Freund
Cancers 2025, 17(4), 681; https://doi.org/10.3390/cancers17040681 - 18 Feb 2025
Viewed by 336
Abstract
Cancer is the second-leading cause of death in developed societies. Specifically, cancers of the spine and brain come with significant therapeutic challenges. Chordomas are semi-malignant tumors that develop from embryonic residuals at the skull base (clival) or coccyx (sacral). Small tumor fragments can [...] Read more.
Cancer is the second-leading cause of death in developed societies. Specifically, cancers of the spine and brain come with significant therapeutic challenges. Chordomas are semi-malignant tumors that develop from embryonic residuals at the skull base (clival) or coccyx (sacral). Small tumor fragments can remain in the operation cavities during surgical resection, forming new tumor sites. This requires repeated surgeries or the application of proton-beam radiation and chemotherapy, which often do not lead to complete remission of the tumors. Hence, there is a need for novel therapeutic avenues that are not limited to killing visible tumors but can be applied after surgery to decrease chordoma recurrences. Reactive oxygen species (ROS) generated locally via novel medical gas plasma technologies are one potential approach to address this clinical problem. Previously, broad-spectrum free radicals generated by these cold physical plasmas operated at about body temperature were shown to oxidize cancer cells to the disadvantage of their growth and induce immunogenic cancer cell death (ICD), ultimately promoting anticancer immunity. This review outlines the clinical challenges of chordoma therapy, how medical gas plasma technology could serve as an adjuvant treatment modality, and potential immune-related mechanisms of action that could extend the longevity of gas plasma therapy beyond its acute local tissue effects. Full article
(This article belongs to the Special Issue Advances in Spine Oncology: Research and Clinical Studies)
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<p>(<b>a</b>) Sites of chordoma occurrence and metastatic foci as well as local compression of tissue of the neuronal axis. (<b>b</b>) Immuno-oncological dimension of chordomas characterized by elevated M2-macrophages and T-cells, as well as expression of immune checkpoints and cytokine secretion. Figure adapted from [<a href="#B5-cancers-17-00681" class="html-bibr">5</a>].</p>
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<p>(<b>a</b>) Ex vivo tissue exposure to the atmospheric pressure medical gas plasma jet kINPen MED was used to generate various ROS types. The principle is based on argon gas being ignited at the jet head’s center by a high-frequency, high-voltage electrode; ionized argon being driven out into the ambient air; and argon ions generating radicals from ambient air oxygen and nitrogen, effectively generating reactive oxygen and nitrogen species (ROS). (<b>b</b>) Optical emission spectroscopy of the gas plasma shows distinct peaks that can be attributed to different reactive species precursors, such as hydrogel radicals (309 nm) and the second positive system of nitrogen. Figure copyright: the authors.</p>
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<p>Principles of immunogenic cell death (ICD), modified after [<a href="#B81-cancers-17-00681" class="html-bibr">81</a>]. Therapy-induced stress, such as through the local delivery of high levels of ROS via medical gas plasma technology, induced an inflammatory type of cell death in chordoma cells, characterized by DAMP release (e.g., ATP) and exposure of pro-phagocytic signals (e.g., calreticulin, CRT), leading to enhanced phagocytosis by antigen-presenting cells (APCs). Activated by DAMPs and ICD, APC maturation and migration into the draining lymph node lead to improved tumor antigen presentation, e.g., to T-cells (TCs), rendering them into effector cells entering the circulation to target tumor cells systemically and condition the tumor microenvironment (TME).</p>
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<p>(<b>a</b>) Sketch of the two aspects of direct tumor toxicity and anticancer immunity that are needed to balance future effective chordoma therapies. (<b>b</b>) MRI images (T1-weighted with contrast medium) show the typical clinical course of an aggressive chordoma (<b>I</b>) at time of diagnosis, (<b>II</b>) after surgical decompression of the brain stem and important neuronal structures, and (<b>III</b>) after its recurrence as extensive disease. (<b>c</b>) Summary of the proposed chordoma treatment that adds gas plasma oxidation after surgical decompression to minimize the risk of tumor recurrence. Figure copyright is by the authors.</p>
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18 pages, 1488 KiB  
Article
Plasma-Induced Abatement of Tar from Syngas Produced in Municipal Waste Gasification: Thermodynamic Modeling with Experimental Validation
by Mobish A. Shaji, Francis Eboh, Alexander Rabinovich, Liran Dor and Alexander Fridman
Plasma 2025, 8(1), 6; https://doi.org/10.3390/plasma8010006 - 16 Feb 2025
Viewed by 185
Abstract
Municipal waste gasification presents a promising avenue to extract useful energy from waste through syngas. This technology’s application is limited by tar formation (long-chain hydrocarbons), which can decrease energy conversion efficiency and applications of raw syngas. Non-thermal plasma-based tar degradation is a simple [...] Read more.
Municipal waste gasification presents a promising avenue to extract useful energy from waste through syngas. This technology’s application is limited by tar formation (long-chain hydrocarbons), which can decrease energy conversion efficiency and applications of raw syngas. Non-thermal plasma-based tar degradation is a simple and cost-effective alternative to existing thermal and catalytic tar mitigation methods. While plasma stimulates tar reformation reactions like steam reformation, there are thermodynamic energy requirements associated with these endothermic processes. Determining thermodynamic energy requirements and the equilibrium composition of products during tar reformation can aid with the proper optimization of the treatment process. In the present study, thermodynamic modeling and experimental validation are conducted to study energy requirements and product formation during the plasma-assisted steam reformation of tar present in raw syngas with an inlet temperature of 300 °C and 30% moisture content. The thermodynamic study evaluated the effect of adding air into the system (to increase the temperature by oxidizing a portion of raw syngas). Results show that up to 75% of energy requirement can be brought down by adding up to 30% air; experimental validation using gliding arc discharge with 30% air addition agrees with the thermodynamic model finding. The thermodynamic model predicted an increase in H2 and CO concentration with the degradation of tar, but experimental validation reported a reduction in H2 and CO concentration with the degradation of tar, as syngas was consumed to increase the temperature to support oxidation, owing to the low temperature (300 °C) and significant moisture presence (~30%) of raw syngas analyzed in this study. Full article
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<p>Schematic of gliding arc discharge-based biomass tar reformation system.</p>
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<p>Thermodynamic modeling results of tar degradation as a function of specific energy input.</p>
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<p>Thermodynamic modeling results showing influence of air addition on the combined molar concentration of hydrogen and carbon monoxide in syngas as a function of specific energy input.</p>
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<p>Tar degradation observed during experimental analysis and thermodynamic modeling as a function of specific energy input. n = 2.</p>
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<p>Changes to <math display="inline"><semantics> <mrow> <msub> <mrow> <mi mathvariant="normal">H</mi> </mrow> <mrow> <mn>2</mn> </mrow> </msub> <mo> </mo> <mi mathvariant="normal">a</mi> <mi mathvariant="normal">n</mi> <mi mathvariant="normal">d</mi> <mo> </mo> <mi mathvariant="normal">C</mi> <mi mathvariant="normal">O</mi> </mrow> </semantics></math> concentration in syngas during experimental analysis and thermodynamic modeling as a function of specific energy input. n = 2.</p>
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<p>%Tar degradation as a function of energy required relative to the heating value of raw syngas. n = 2.</p>
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12 pages, 578 KiB  
Systematic Review
Hemolytic Anemia Linked to Epstein–Barr Virus Infectious Mononucleosis: A Systematic Review of the Literature
by Dario F. Meloni, Pietro B. Faré, Gregorio P. Milani, Sebastiano A. G. Lava, Mario G. Bianchetti, Samuele Renzi, Massimiliano Bertacchi, Lisa Kottanattu, Gabriel Bronz and Pietro Camozzi
J. Clin. Med. 2025, 14(4), 1283; https://doi.org/10.3390/jcm14041283 - 15 Feb 2025
Viewed by 185
Abstract
Background: In Epstein–Barr virus infectious mononucleosis, hemolytic anemia occasionally occurs. Methods: To characterize hemolytic anemia linked to Epstein–Barr virus infectious mononucleosis, we performed a systematic review (PROSPERO CRD42024597183) in the United States National Library of Medicine, Excerpta Medica, and Web of [...] Read more.
Background: In Epstein–Barr virus infectious mononucleosis, hemolytic anemia occasionally occurs. Methods: To characterize hemolytic anemia linked to Epstein–Barr virus infectious mononucleosis, we performed a systematic review (PROSPERO CRD42024597183) in the United States National Library of Medicine, Excerpta Medica, and Web of Science with no restrictions on language. Only reports published since 1970 were included. Eligible were reports describing hemolytic anemia in subjects with clinical signs and microbiological markers of Epstein–Barr virus mononucleosis. Results: In the literature, we detected 56 reports released between 1973 and 2024, documenting 60 individuals (32 females and 28 males; 27 children and 33 adults) with hemolytic anemia linked to Epstein–Barr virus infectious mononucleosis. The mechanism underlying anemia was categorized as cold-antibody-mediated (N = 31; 52%), warm-antibody-mediated (N = 18, 30%), mixed warm- and cold-antibody-mediated (N = 4; 6.7%), or paroxysmal cold hemoglobinuria (N = 2; 3.3%). The remaining 5 cases (8.3%) remained unclassified. Observation alone was the chosen approach in 23% of cases (N = 14). Steroids (67%; N = 40) and blood transfusions (38%; N = 23) were the most commonly used treatment, while plasma exchange, intravenous polyclonal immunoglobulin, rituximab, and splenectomy were used less frequently. Observation was slightly but significantly (p = 0.032) more common in cases of cold-antibody-mediated anemia compared to all other cases combined. Patients recovered a median of 28 [interquartile range 21–39] days after disease onset. Two patients with warm-antibody-mediated hemolytic anemia died. Conclusions: This literature review points out that Epstein–Barr virus, like Mycoplasma pneumoniae, cytomegalovirus, or severe acute respiratory syndrome coronavirus 2, may act as a trigger for immune-mediated hemolytic anemia. Full article
(This article belongs to the Section Infectious Diseases)
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<p>Hemolytic anemia linked to Epstein–Barr virus infectious mononucleosis. Flowchart of the literature search. No new pertinent reports were identified in the literature search updated on 20 December 2024.</p>
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18 pages, 2377 KiB  
Article
Regulation of NO-Generating System Activity in Cucumber Root Response to Cold
by Małgorzata Reda, Katarzyna Kabała, Jan Stanisławski, Kacper Szczepski and Małgorzata Janicka
Int. J. Mol. Sci. 2025, 26(4), 1599; https://doi.org/10.3390/ijms26041599 - 13 Feb 2025
Viewed by 301
Abstract
Nitric oxide (NO) functions as a signaling molecule in plant adaptation to changing environmental conditions. NO levels were found to increase in plants in response to low temperatures (LTs). However, knowledge of the pathways involved in enhanced NO production under cold stress is [...] Read more.
Nitric oxide (NO) functions as a signaling molecule in plant adaptation to changing environmental conditions. NO levels were found to increase in plants in response to low temperatures (LTs). However, knowledge of the pathways involved in enhanced NO production under cold stress is still limited. For this reason, we aimed to determine the role of different NO sources in NO generation in cucumber roots exposed to 10 °C for short (1 d) and long (6 d) periods. The short-term treatment of seedlings with LT markedly increased plasma membrane-bound nitrate reductase (PM-NR) activity and induced the expression of three genes encoding NR in cucumber (CsNR1-3). On the other hand, long-term exposure was related to both increased cytoplasmic NR (cNR) activity and induced expression of the CsARC gene, encoding the amidoxime-reducing component (ARC) protein. The decrease in nitrite reductase (NiR) activity and the higher NO2/NO3 ratio in the roots of plants exposed to LTs for 1 d suggest that tissue conditions may favor NR-dependent NO production. Regardless of NR stimulation, a significant increase in NOS-like activity was observed in the roots, especially during the long-term treatment of plants with LT. These results indicate that diverse NO-producing routes, both reductive and oxidative, are activated in cucumber tissues at different stages of cold stress. Full article
(This article belongs to the Section Molecular Plant Sciences)
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<p>Bio-imaging of NO production (<b>A</b>) and mean DAF-2D-related fluorescence density (<b>B</b>) in root apical segments of cucumber seedlings not treated (control) and treated with LT for 1 day (LT 1 d) and 6 days (LT 6 d). Some sets of plants were incubated in the presence of an NR inhibitor (0.1 mM sodium tungstate; +WO<sub>4</sub><sup>2−</sup>) or a mammalian NOS inhibitor (0.1 mM AET, +AET). Bio-imaging of NO generation was monitored by labeling with the NO-specific fluorescent dye, DAF-2D, and imaged using fluorescence microscopy. The images shown in (<b>A</b>) are representative for at least 3–5 roots for each treatment from 3 independent replications of the experiment. The asterisks in (<b>B</b>) indicate a difference in comparison to the control (Student’s <span class="html-italic">t</span>-test, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Total activity, actual activity, and enzyme activation state of cNR (<b>A</b>, <b>B</b> and <b>C</b>, respectively) and PM-NR (<b>D</b>, <b>E</b> and <b>F</b>, respectively) in roots of cucumber seedlings not treated (control) and treated with LT for 1 day (LT 1 d) and 6 days (LT 6 d). Further, 326.96 ± 40.1 (<b>A</b>) and 98.30 ± 15.05 (<b>B</b>) nmol NO<sub>2</sub><sup>−</sup> × g FW<sup>−1</sup> × h<sup>−1</sup> were used as 100% for cNR. Additionally, 223.22 ± 59.16 (<b>D</b>) and 155.01 ± 35.01 (<b>E</b>) nmol NO<sub>2</sub><sup>−</sup> × mg<sup>−1</sup> protein × h<sup>−1</sup> were used as 100% for PM-NR. All results are means ± SDs of 3 replications of 4 independent experiments. The asterisks indicate significant differences in comparison to the control (Student’s <span class="html-italic">t</span>-test, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Relative expression of CsNR1-3 genes in roots of cucumber seedlings not treated (control) and treated with LT for 1 day (LT 1 d) and 6 days (LT 6 d). Expression was determined using real-time PCR. All results were normalized to the reference expression of CsTIP41 encoding the TIP41-like protein. The values presented are means ± SDs from 3 replicates of 3 independent experiments. The asterisks indicate significant differences compared to the control (Student’s <span class="html-italic">t</span>-test, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Nitrite reductase (NiR) activity in roots of cucumber seedlings not treated (control) and treated with LT for 1 day (LT 1 d) and 6 days (LT 6 d). The data presented are the means of 3 replications of 5 independent experiments. Error bars represent SDs. Asterisks indicate significant differences compared to the control (Student’s <span class="html-italic">t</span>-test, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>NOS-like activity measured as arginine-dependent NADPH oxidation in extracts prepared from roots of cucumber seedlings not treated (control) and treated with LT for 1 d (LT 1 d) and 6 d (LT 6 d). Data presented are means of 3 replicates of 3 independent experiments, with error bars representing the SD. Asterisks indicate significant differences compared to the control (Student’s <span class="html-italic">t</span>-test, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Relative expression of <span class="html-italic">CsARC</span> in the roots of cucumber seedlings not treated (control) and treated with LT for 1 d (LT 1 d) and 6 d (LT 6 d). Expression was determined by real-time PCR. The results were normalized to the reference expression of <span class="html-italic">CsCACS</span>, which encodes a clathrin adaptor complex subunit. Presented values are means ± SDs of 3 replicates of 3 independent experiments. Asterisks indicate significant differences compared to the control (Student’s <span class="html-italic">t</span>-test, <span class="html-italic">p</span> &lt; 0.05).</p>
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40 pages, 12596 KiB  
Review
A Review on the Additive Manufacturing of W-Cu Composites
by Muhammad Hussain, Bosheng Dong, Zhijun Qiu, Ulf Garbe, Zengxi Pan and Huijun Li
Metals 2025, 15(2), 197; https://doi.org/10.3390/met15020197 - 13 Feb 2025
Viewed by 371
Abstract
In recent years, W-Cu composite systems have become very interesting subjects due to good electrical and thermal conductivity, high-temperature strength, certain plasticity, and excellent radiation resistance. W-Cu composites are a very important class of materials in applications like PFM (plasma facing materials), functional [...] Read more.
In recent years, W-Cu composite systems have become very interesting subjects due to good electrical and thermal conductivity, high-temperature strength, certain plasticity, and excellent radiation resistance. W-Cu composites are a very important class of materials in applications like PFM (plasma facing materials), functional graded materials (FGM), electronic packaging materials, high-voltage electrical contacts, sweating materials, shaped charge liners, electromagnetic gun-rail materials, kinetic energy penetrators, and radiation shielding/protection. There is no possibility of forming a crystalline structure between these two materials. However, due to the unique properties these materials possess, they can be used by preparing them as a composite. Generally, W-Cu composites are prepared via the conventional powder metallurgy routes, i.e., sintering, hot pressing, hot isostatic pressing, isostatic cold pressing, sintering and infiltration, and microwave sintering. However, these processes have certain limitations, like the inability to produce bulk material, they are expensive, and their adoptability is limited. Here, in this review, we will discuss in detail the fabrication routes of additive manufacturing, and its current progress, challenges, trends, and associated properties obtained. We will also explain the challenges for the additive manufacturing of the composite. We will also compare W-Cu composites to other materials that can challenge them in terms of specific applications or service conditions. The solidification mechanism will be explained for W-Cu composites in additive manufacturing. Finally, we will conclude the progress of additive manufacturing of W-Cu composites to date and suggest future recommendations based on the current challenges in additive manufacturing. Full article
(This article belongs to the Section Welding and Joining)
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<p>Phase diagram of W and Cu. Reprinted with permission from Ref. [<a href="#B45-metals-15-00197" class="html-bibr">45</a>].</p>
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<p>Gibbs free energy vs. alloy composition, Reprinted with permission from Ref. [<a href="#B46-metals-15-00197" class="html-bibr">46</a>]. 1985, AIP Publishing.</p>
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<p>Formation enthalpy calculated vs. compositional ratios using MD simulations, reprinted with permission from ref. [<a href="#B48-metals-15-00197" class="html-bibr">48</a>], 2017, Elsevier.</p>
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<p>Surface energy of different materials studied in the research, reprinted from [<a href="#B49-metals-15-00197" class="html-bibr">49</a>].</p>
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<p>Surface energy of different transition metals and W (The dotted line shows results based on FCD-LMTO calculations), reprinted with permission from ref. [<a href="#B50-metals-15-00197" class="html-bibr">50</a>], 2018, Elsevier.</p>
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<p>Effect of nano-sized particles on the formation enthalpy for W50Cu50 system, reprinted from Ref. [<a href="#B47-metals-15-00197" class="html-bibr">47</a>].</p>
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<p>(<b>a</b>) Discontinuous FGM and (<b>b</b>) continuous FGM, (<b>c</b>–<b>e</b>) schematic diagrams showing discontinuous FGMs that contain interfaces with gradual change in composition, grain orientation and volume fractions of two types of second-phase particles, respectively. (<b>f</b>–<b>h</b>) schematic diagrams showing continuous FGMs in absence of interfaces and with gradual change in grain size, fiber orientation and volume fraction of second-phase particles. Reprinted with permission from Ref. [<a href="#B62-metals-15-00197" class="html-bibr">62</a>], 2019, Elsevier.</p>
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<p>Shift (red arrow) in Ductile-to-brittle transition temperature (DBTT) of W tested in cold-rolled, hot-rolled, and annealed conditions in LS and TS orientations. Reprinted with permission from Ref. [<a href="#B65-metals-15-00197" class="html-bibr">65</a>], 2016, Elsevier.</p>
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<p>Effect of Re addition on DBTT in W, reprinted with permission from Ref. [<a href="#B66-metals-15-00197" class="html-bibr">66</a>], 2018, Elsevier.</p>
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<p>Mechanistic models of heat and mass transfer and fluid flow in AM. reprinted with permission from Ref. [<a href="#B70-metals-15-00197" class="html-bibr">70</a>].</p>
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<p>(<b>a</b>) Infiltration method for W-Cu production. (<b>b</b>) W-Cu microstructure obtained, reprinted with permission from Ref. [<a href="#B25-metals-15-00197" class="html-bibr">25</a>], 2018, Elsevier.</p>
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<p>Liquid-phase sintering mechanism, reprinted with permission from Ref. [<a href="#B78-metals-15-00197" class="html-bibr">78</a>], 2009, Springer Nature.</p>
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<p>(<b>a</b>) Vacuum and (<b>b</b>) microwave heating (the intensity of color represents the heat energy), reprinted with permission from Ref. [<a href="#B82-metals-15-00197" class="html-bibr">82</a>], 2024, Wiley.</p>
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<p>Schematic of hot-pressing process, reprinted with permission from Ref. [<a href="#B60-metals-15-00197" class="html-bibr">60</a>], 2008, Elsevier.</p>
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<p>Spark plasma sintering setup and the heating mechanism, reprinted with permission from Ref. [<a href="#B82-metals-15-00197" class="html-bibr">82</a>], 2024, Wiley.</p>
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<p>Schematic of injection molding, reprinted with permission from Ref. [<a href="#B25-metals-15-00197" class="html-bibr">25</a>], 2018, Elsevier.</p>
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<p>Schematic of mechanical alloying: (<b>a</b>) process and (<b>b</b>) mechanism. Reprinted with permission from Ref. [<a href="#B92-metals-15-00197" class="html-bibr">92</a>], 2013, Elsevier.</p>
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<p>A schematic of fabrication of W-Cu composite using laser technology, reprinted with permission from Ref. [<a href="#B93-metals-15-00197" class="html-bibr">93</a>], 2020, Elsevier.</p>
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<p>W-Cu composite prepared by laser additive manufacturing technology with different scanning and laser-power levels, reprinted with permission from Ref. [<a href="#B94-metals-15-00197" class="html-bibr">94</a>], 2018, Elsevier.</p>
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<p>Schematic of electron beam melting, reprinted from Ref. [<a href="#B96-metals-15-00197" class="html-bibr">96</a>].</p>
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<p>SEM surface morphology of the W-(Ni)-Cu composites. (<b>a</b>) W-Cu40 wt.%; (<b>b</b>) W-Cu30 wt.%; (<b>c</b>) W-Cu25 wt.%; and (<b>d</b>) W-Ni5 wt.%–Cu15 wt.%, reprinted with permission from Ref. [<a href="#B42-metals-15-00197" class="html-bibr">42</a>], 2016, Elsevier.</p>
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<p>Surface SEM images of Cu-15W sample before and after HCPEB irradiation. (<b>a</b>) Initial, (<b>b</b>) 5 pulses, (<b>c</b>) 10 pulses, and (<b>d</b>) BSE image of 10-pulsed sample, reprinted with permission from Ref. [<a href="#B43-metals-15-00197" class="html-bibr">43</a>], 2018, Elsevier.</p>
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<p>Cross-sections of three-layer W–Cu cladding on a steel substrate: (<b>a</b>) one layer (75% W), (<b>b</b>) two layers (75% + 95% W), and (<b>c</b>) three layers (75% + 95% + 98% W), reprinted from Ref. [<a href="#B44-metals-15-00197" class="html-bibr">44</a>].</p>
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<p>Typical (<b>a</b>,<b>b</b>) microstructure and (<b>c</b>) XRD of W and Cu system, reprinted with permission from Ref. [<a href="#B19-metals-15-00197" class="html-bibr">19</a>], 2022, Elsevier.</p>
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<p>TEM observations at W/Cu interface: (<b>a</b>) HAADF image; (<b>b</b>) EDX line-scanning profile along the red arrow marked in (<b>a</b>); (<b>c</b>,<b>d</b>) element mapping; (<b>e</b>) HAADF image of the W/Cu interface; and (<b>f</b>) HR-TEM image of the interface, reprinted with permission from Ref. [<a href="#B151-metals-15-00197" class="html-bibr">151</a>], 2022, Elsevier.</p>
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<p>Colling rate on (<b>a</b>) Cu substrate; and (<b>b</b>) Cu, Al, and SS substrate, reprinted with permission from Ref. [<a href="#B172-metals-15-00197" class="html-bibr">172</a>], 1995, Elsevier.</p>
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<p>Schematic of laser welding and molten pool formation, reprinted from Ref. [<a href="#B178-metals-15-00197" class="html-bibr">178</a>].</p>
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<p>Schematic of electron beam melting, reprinted with permission from Ref. [<a href="#B179-metals-15-00197" class="html-bibr">179</a>], 2016, Elsevier.</p>
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<p>(<b>a</b>) Schematic of the welding principle, (<b>b</b>) Material states, fluid flow and four types of forces involved during the welding process, reprinted from Ref. [<a href="#B181-metals-15-00197" class="html-bibr">181</a>].</p>
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<p>The degree of undercooling explained (<b>a</b>) Hypocooling, (<b>b</b>) Critical undercooling, (<b>c</b>) Hypercooling, reprinted with permission from Ref. [<a href="#B185-metals-15-00197" class="html-bibr">185</a>], 2010, Springer.</p>
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<p>Geometrically necessary dislocations (GNDs) and statistically stored dislocations (SSDs), reprinted with permission from Ref. [<a href="#B187-metals-15-00197" class="html-bibr">187</a>], 2019, Elsevier.</p>
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<p>Solidification mechanism of W-Cu system explained in four steps, reprinted with permission from Ref. [<a href="#B19-metals-15-00197" class="html-bibr">19</a>], 2022, Elsevier.</p>
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20 pages, 1470 KiB  
Review
Cold Plasma-Induced Modulation of Protein and Lipid Macromolecules: A Review
by Srutee Rout, Pradeep Kumar Panda, Pranjyan Dash, Prem Prakash Srivastav and Chien-Te Hsieh
Int. J. Mol. Sci. 2025, 26(4), 1564; https://doi.org/10.3390/ijms26041564 - 13 Feb 2025
Viewed by 597
Abstract
Nowadays, the food industry is prioritizing many innovative processing technologies that can produce minimally processed foods with superior and higher quality, lower costs, and faster operations. Among these advancements, cold plasma (CP) processing stands out for its remarkable capabilities in food preservation and [...] Read more.
Nowadays, the food industry is prioritizing many innovative processing technologies that can produce minimally processed foods with superior and higher quality, lower costs, and faster operations. Among these advancements, cold plasma (CP) processing stands out for its remarkable capabilities in food preservation and extending the shelf life. Beyond its established role in microbial inactivation, CP has emerged as a transformative tool for modifying food biomolecules through reactive plasma species, addressing the versatile requirements of food industries for various applications. This review focuses on the interactions between reactive plasma species and essential food macromolecules, including proteins, lipids, and polysaccharides. The novelty lies in its detailed examination of how CP technology triggers structural, functional, and biochemical changes in proteins and lipids and explains the mechanisms involved. It connects fundamental molecular transformations to practical applications, such as enhanced protein functionality, lipid stabilization, and improved oxidative resistance. CP induces alterations in protein structure, especially in amino acid configurations, that can be applicable to the formulation of advanced gel, 3D printing, thermostable emulsions, enhanced solubility, and sensory materials. This review explores the ability of CP to modify protein allergenicity, its different effects on the mechanical and interfacial properties of proteins, and its role in the production of trans-fat-free oils. Despite its potential, a detailed understanding of the mechanism of CP’s interactions with food macromolecules is also discussed. Furthermore, this review addresses key challenges and outlines future research opportunities, positioning CP as a sustainable and adaptable approach for innovating next-generation food systems. Further research is crucial to fully understand the potential of CP for food processing, followed by product development. Full article
(This article belongs to the Section Macromolecules)
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<p>Schematic diagram showing generation of plasma by (<b>a</b>) radiofrequency, (<b>b</b>) dielectric barrier discharge, (<b>c</b>) surface DBD, (<b>d</b>) corona discharge, (<b>e</b>) microwave, (<b>f</b>) plasma jet, and (<b>g</b>) gliding arc discharge—adapted, redrawn, and modified from ref. [<a href="#B32-ijms-26-01564" class="html-bibr">32</a>] with copyright permission from Elsevier publication.</p>
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<p>Schematic diagram modification of zeolite by the DBD in the CO<sub>2</sub> atmosphere—adapted from ref. [<a href="#B55-ijms-26-01564" class="html-bibr">55</a>] with copyright permission from Wiley publication.</p>
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<p>CP technique’s effect on proteins—adapted from ref. [<a href="#B80-ijms-26-01564" class="html-bibr">80</a>] with copyright permission from Elsevier publication.</p>
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20 pages, 1096 KiB  
Review
A Review on the Conservation of South African Indigenous Poultry Breeds: A Focus on Semen Cryopreservation
by Rantloko Rolly Maapola, Jabulani Nkululeko Ngcobo, Khathutshelo Agree Nephawe, Tshimangadzo Lucky Nedambale and Fhulufhelo Vincent Ramukhithi
Animals 2025, 15(4), 529; https://doi.org/10.3390/ani15040529 - 12 Feb 2025
Viewed by 369
Abstract
Understanding the genetic, physiological, and nutritional characteristics of native chickens in South Africa has been significantly hindered by studies over the last ten years. These chickens hold significant economic, social, and cultural importance for South African communities, particularly those marginalized. Despite their reputation [...] Read more.
Understanding the genetic, physiological, and nutritional characteristics of native chickens in South Africa has been significantly hindered by studies over the last ten years. These chickens hold significant economic, social, and cultural importance for South African communities, particularly those marginalized. Despite their reputation for lower egg productivity, they are highly valued for their flavorful meat by consumers. Many local chicken ecotypes and breeds remain undocumented and in danger of going extinct, even though some have been classified. To tackle this issue, the Food and Agriculture Organization has launched an indigenous poultry conservation program. One crucial method employed is assisted reproductive biotechnologies such as cryopreservation, which serves as an ex situ conservation strategy for preserving the germplasm of endangered animals. In avian species, cryopreservation is particularly beneficial for the long-term storage of sperm cells, although it necessitates the use of cryoprotectants to shield sperm cells from cold shock during freezing. However, the use of cryoprotectants can lead to thermal shocks that may damage the sperm cell plasma membrane, potentially reducing viability and fertility. Furthermore, the membranes of avian sperm cells are highly polyunsaturated fatty acids, which can undergo lipid peroxidation (LPO) when reactive oxygen species (ROS) are present. This review focuses on current knowledge and the latest effective strategies for utilizing cryopreservation to conserve semen from indigenous poultry breeds. Full article
(This article belongs to the Section Poultry)
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<p>Schematic representation of conservation strategies. Source: [<a href="#B37-animals-15-00529" class="html-bibr">37</a>].</p>
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<p>(<b>A</b>) Venda; (<b>B</b>) Boschveld; (<b>C</b>) Potchefstroom Koekoek; (<b>D</b>) Naked Neck; and (<b>E</b>) Ovambo. Source: [<a href="#B6-animals-15-00529" class="html-bibr">6</a>].</p>
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15 pages, 3032 KiB  
Article
Antitumor Potential of Different Treatment Approaches Using Cold Atmospheric Pressure Plasma on Oral Squamous Cell Carcinoma Models: In Vitro Study
by Ognjan Pavlović, Miloš Lazarević, Aleksandar Jakovljević, Nikola Škoro, Nevena Puač, Slavko Mojsilović and Maja Miletić
Biomedicines 2025, 13(2), 443; https://doi.org/10.3390/biomedicines13020443 - 11 Feb 2025
Viewed by 601
Abstract
Background/Objectives: Cold atmospheric plasma (CAP) has shown a strong anticancer effect on a variety of tumors, presenting a new approach for the effective treatment of oral squamous cell carcinoma (OSCC), one of the most prevalent malignant neoplasms with a high mortality rate. Here, [...] Read more.
Background/Objectives: Cold atmospheric plasma (CAP) has shown a strong anticancer effect on a variety of tumors, presenting a new approach for the effective treatment of oral squamous cell carcinoma (OSCC), one of the most prevalent malignant neoplasms with a high mortality rate. Here, we aimed to comprehensively investigate the antitumor potential of two approaches of CAP treatment on both two-dimensional and three-dimensional OSCC cell line models, as well as to analyze whether plasma treatment enhances the sensitivity of OSCC to chemotherapy. Methods: An in-house designed plasma needle, with helium as a working gas, was used to treat the SCC-25 cell line directly or indirectly via plasma-treated medium (PTM). The antitumor effect of CAP was assessed by measuring cell viability, apoptosis, adhesion, and migration. In addition, the combined effect of PTM and cisplatin was analyzed in SCC-25 tumor spheroids, as a more complex and reliable in vitro model. Results: Both plasma treatments showed time-dependent antitumor effects affecting their viability, adhesion, and migration. The rate of apoptosis was higher after incubation with PTM and is mediated by the intrinsic pathway. By utilizing the 3D spheroid carcinoma model, we confirmed the antitumor potential of CAP and additionally demonstrated an increased chemosensitivity of PTM-treated carcinoma cells. Conclusions: The results of our study illustrate a promising avenue for the application of CAP as a therapeutic option for OSCC, either as a standalone treatment or in combination with cisplatin. Full article
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<p>Positioning of the plasma needle in (<b>A</b>) direct treatments and (<b>B</b>) treatment of cell culture medium (PTM generation). (<b>C</b>) Photography of the plasma treating cell culture medium.</p>
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<p>MTT assay results. The values are presented as the percentage of viable cells (normalized ratio of treated and control samples values). (<b>A</b>) Direct plasma treatment for 10 s, 30 s, and 60 s; (<b>B</b>) indirect plasma treatment (PTM exposed to CAP for 30 s, 60 s, and 120 s). The control group present the untreated cells grown in standard medium. The data are presented as mean ± SD, and statistical significance is denoted as * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01; **** <span class="html-italic">p</span> &lt; 0.0001.</p>
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<p>Flow cytometric screening of SCC-25 cells. PE Annexin-V staining was used to measure the apoptosis rate immediately after direct CAP treatment and 24 h after the incubation with PTM. (<b>A</b>) Stacked column diagram shows mean values of live vs. early apoptotic vs. late apoptotic/necrotic cell populations after different treatment protocols independently repeated three times. (<b>B</b>–<b>F</b>) The representative density plots for each treatment group are presented: (<b>B</b>) control untreated cells (direct treatment control); (<b>C</b>) cells treated directly with CAP; (<b>D</b>) cells incubated in control medium (indirect treatment control); (<b>E</b>) cells incubated with PTM; (<b>F</b>) cells treated with 100 μM of cisplatin (positive control).</p>
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<p>Caspase activation luminescence-based assay. The activation of different caspase systems (caspase 3/7, caspase 9, caspase 8) was measured 24 h after the incubation of SCC-25 cells with PTM obtained by 120 s exposure to CAP. Control cells were incubated in standard medium; positive control cells were treated with 100 μM of cisplatin. The data are presented as mean ± SD, and statistical significance is denoted as * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Adhesion assay results. The values are presented as percentages of adherent cells (normalized ratio of treated samples values with values of corresponding controls). (<b>A</b>) Direct CAP treatment; (<b>B</b>) indirect treatment with PTM; the data are presented as mean ± SD of three repeated experiments, and statistical significance is denoted as * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001; **** <span class="html-italic">p</span> &lt; 0.0001.</p>
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<p>Wound healing assay results. (<b>A</b>) Direct CAP treatment; (<b>B</b>) indirect treatment with PTM; the data are presented as mean ± SD, and statistical significance is denoted as ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001; **** <span class="html-italic">p</span> &lt; 0.0001.</p>
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<p>Cell titer Glo<sup>®</sup> assay results in 3D spheroid models. The viability was measured after 24 h of incubation with PTM. (<b>A</b>) PTM treated for 120 s, alone or with increasing concentrations of cisplatin (0–100 μM); (<b>B</b>) PTM treated for 180 s, alone or with increasing concentrations of cisplatin (0–100 μM).</p>
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17 pages, 5133 KiB  
Article
Investigating the Synergistic Bactericidal Effects of Cold Plasma and Ultraviolet Radiation on Pseudomonas fragi
by Haidu Yuan, Fei Chen, Jiajia Zhang, Xinglei Guo, Jianhao Zhang and Wenjing Yan
Foods 2025, 14(4), 550; https://doi.org/10.3390/foods14040550 - 7 Feb 2025
Viewed by 435
Abstract
Cold plasma is a novel non-thermal processing technology with broad application prospects in food preservation. When combined with other physical sterilization technologies, it enhances sterilization efficiency and broadens its application scope, providing a safe and effective alternative to traditional sterilization methods. In this [...] Read more.
Cold plasma is a novel non-thermal processing technology with broad application prospects in food preservation. When combined with other physical sterilization technologies, it enhances sterilization efficiency and broadens its application scope, providing a safe and effective alternative to traditional sterilization methods. In this paper, the sterilization effect of surface dielectric barrier discharge (SDBD) plasma combined with 222 nm ultraviolet (UV) irradiation against Pseudomonas fragi (P. fragi) was explored for the first time. The sterilization process parameters of SDBD + UV were optimized using the response surface methodology. And the sterilization mechanism of SDBD + UV was preliminary elucidated. The results indicated that the SDBD + UV treatment was highly effective against P. fragi. It could eliminate 6.35 Log CFU/g of P. fragi within 150 s, establishing optimal sterilization parameters: a radiation distance of 16.4 cm and a saving time (a period of preservation in which the samples were retained in the device after the treatment) of 120 s. Furthermore, the treatment caused significant damage to the cell membrane of P. fragi, leading to membrane perforation and content leakage. It also induced oxidative stress, as evidenced by membrane lipid peroxidation, alterations in intracellular reactive oxygen species (ROS) content, and a decrease in antioxidant enzyme activity. This study provides a theoretical basis for the application of cold plasma combined with 222 nm UV treatment in the meat industry. Full article
(This article belongs to the Special Issue Antimicrobial Strategies in Food Processing, Production and Storage)
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<p>Schematic diagram of the SDBD + UV treatment device.</p>
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<p>Single-factor experiment effects of three independent variables on the antibacterial activity of SDBD + UV: (<b>A</b>) treatment time (irradiation distance: 18 cm, saving time: 90 s); (<b>B</b>) irradiation distance (treatment time: 90 s, saving time: 90 s); (<b>C</b>) saving time (treatment time: 90 s, irradiation distance: 18 cm). Different letters in the figure indicate significant differences between the data (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>(<b>A</b>) Response surface and (<b>B</b>) contour plots showing the interactive effects of irradiation distance and saving time on the reduction in <span class="html-italic">P. fragi</span> (the fixed treatment time: 90 s).</p>
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<p>The bactericidal effects of SDBD, UV, and SDBD + UV treatments on <span class="html-italic">P. fragi</span>. Different letters in the figure indicate significant differences between the data (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The OES diagram of the SDBD, UV, and SDBD + UV treatments, ranging from 200 to 1150 nm.</p>
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<p>SEM images of <span class="html-italic">P. fragi</span> after different treatments: (<b>A</b>) control group; (<b>B</b>) UV treatment; (<b>C</b>) SDBD treatment; (<b>D</b>) SDBD + UV treatment.</p>
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<p>CLSM images of <span class="html-italic">P. fragi</span> after different treatments followed by staining with PI dye: (<b>A</b>) control group; (<b>B</b>) UV treatment; (<b>C</b>) SDBD treatment; (<b>D</b>) SDBD + UV treatment.</p>
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<p>Effects of SDBD + UV treatment on (<b>A</b>) protein content, leakage of (<b>B</b>) protein, (<b>C</b>) nucleic acid, and (<b>D</b>) potassium leakage of <span class="html-italic">P. fragi</span> over the period of 0–150s. Different letters in the figure indicate significant differences between the data (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>(<b>A</b>) ROS content, (<b>B</b>) membrane lipid peroxidation, and (<b>C</b>) SOD and (<b>D</b>) CAT enzyme activity of <span class="html-italic">P. fragi</span> after SDBD + UV treatment over the period of 0–150 s. Different letters in the figure indicate significant differences between the data (<span class="html-italic">p</span> &lt; 0.05).</p>
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12 pages, 501 KiB  
Article
Passion and Health: How Winter Swimming Influences Blood Morphology and Rheology
by Aneta Teległów, Marta Frankiewicz and Jakub Marchewka
Appl. Sci. 2025, 15(3), 1514; https://doi.org/10.3390/app15031514 - 2 Feb 2025
Viewed by 657
Abstract
An important area of health is health promotion. A healthy lifestyle supports health improvement and early prevention of chronic diseases. Stimulation of the body by cold water swimming and swimming in a swimming pool can lead to adaptive changes beneficial for the human [...] Read more.
An important area of health is health promotion. A healthy lifestyle supports health improvement and early prevention of chronic diseases. Stimulation of the body by cold water swimming and swimming in a swimming pool can lead to adaptive changes beneficial for the human cardiovascular system. Within the winter swimming season of 2023/2024, for a period of 5 months, from November to March, once a week, study participants (n = 30; n = 15 females and n = 15 males) from the Krakow Society of Winter Swimmers ‘Kaloryfer’ in Krakow (Poland) practiced winter swimming in cold water (4–5 °C) and swam in the sports pool of the University of Physical Culture in Krakow in water at a temperature of 28 °C. After a full season of winter swimming and swimming pool sessions, both males and females exhibited a tendency towards lower erythrocyte (p = 0.002), leukocyte (p < 0.001), and platelet counts (p < 0.001), as well as an increase in blood plasma viscosity (within normal limits) (p = 0.001), without any changes in blood aggregation or fibrinogen indicators. The remaining morphological indicators and the elongation index demonstrated only limited variation. Winter swimming induces positive changes in blood morphology and rheology. Full article
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<p>CONSORT flow diagram of study participants.</p>
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11 pages, 3338 KiB  
Technical Note
Chemical Analysis of Plasma-Activated Culture Media by Ion Chromatography
by Marcello Locatelli, Miryam Perrucci, Marwa Balaha, Tirtha-Raj Acharya, Nagendra-Kumar Kaushik, Eun-Ha Choi, Monica Rapino and Vittoria Perrotti
Pharmaceuticals 2025, 18(2), 199; https://doi.org/10.3390/ph18020199 - 1 Feb 2025
Viewed by 577
Abstract
Background: Currently, the procedures and methods applied in biological and medical fields for the determination of reactive oxygen and nitrogen species (RONS), primarily rely on spectrophotometric techniques, which involve the use of colorimetric reagents. While these methods are widely accepted, they exhibit [...] Read more.
Background: Currently, the procedures and methods applied in biological and medical fields for the determination of reactive oxygen and nitrogen species (RONS), primarily rely on spectrophotometric techniques, which involve the use of colorimetric reagents. While these methods are widely accepted, they exhibit significant limitations from an analytical standpoint, particularly due to potential inaccuracies, artifacts, and pronounced susceptibility to matrix effects. The purpose of this Technical Note is to demonstrate the application of ion chromatography—a robust and well-established analytical technique—for the quantification of RONS produced in cell culture media through the exposure to cold atmospheric plasma (CAP), an innovative therapeutic approach for cancer treatment, known as CAP indirect treatment. In addition, the present protocol proposes to apply the pharmacokinetics principles to the RONS generated in plasma-treated liquids (PTLs) following CAP exposure. Methods: The strategy involves elucidating the kinetic profiles of certain characteristic species by evaluating their half-life in the specific media used for cell cultures and investigating their “pharmacokinetic” (PK) profile. In this approach the drug dose is represented by the plasma power and the infusion time corresponds to the exposure time of the culture medium to CAP. Volume-dependent results were shown, focusing on nitrites and nitrates activities, justifying cellular inhibition. Results: This methodology enables the correlation of the PTL biological effects on different cell lines with the PK profiles (dose/time) obtained via ion chromatography. Conclusions: In conclusion, being a simple and green method, it could be used as an alternative to toxic reactions and analytical techniques with higher detection limits, while achieving good resolution. Full article
(This article belongs to the Section Pharmacology)
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<p>Chromatogram obtained in analyzing IC media without plasma activation. (<b>A</b>): DMEM 1:1; (<b>B</b>): McCOY 1:1; (<b>C</b>): RPMI 1:1; (<b>D</b>): DMEM 1:1000; (<b>E</b>): McCOY considering retention time (and stdev) of analytes of interest; (<b>F</b>): RPMI considering retention time (and stdev) of analytes of interest; the rectangles represent each analyte= 1: fluoride, 2: chlorite; 3: bromate, 4: chloride, 5: nitrite, 6: bromide, 7: chlorate, 8: nitrate, 9: phosphate, 10: sulfate.</p>
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<p>Representation of the instrument used: 1: mobile phase; 2: pump that picks up and pushes mobile phase; 3: injection valve; 4: autosampler with vials containing samples; 5: stationary phase; 6: conductivity detector; 7: computer with Chromeleon.</p>
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<p>Simplified illustration of plasma-treated liquid (PTL) production using (<b>a</b>) 2 mL and (<b>b</b>) 5 mL of media.</p>
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<p>Schematic representation of the protocol reported and the chromatogram obtained from analysis.</p>
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19 pages, 9595 KiB  
Article
Plasma-Activated Water Improve Wound Healing in Diabetic Rats by Influencing the Inflammatory and Remodelling Phase
by Jovana Rajić, Nevena Grdović, Anđelija Marković, Nikola Škoro, Svetlana Dinić, Aleksandra Uskoković, Jelena Arambašić Jovanović, Marija Đorđević, Ana Sarić, Melita Vidaković, Nevena Puač and Mirjana Mihailović
Int. J. Mol. Sci. 2025, 26(3), 1265; https://doi.org/10.3390/ijms26031265 - 31 Jan 2025
Viewed by 601
Abstract
Diabetic foot ulcers have an enormous impact on patients’ quality of life and represent a major economic burden. The cause is delayed and incomplete wound healing due to hyperglycemia, reduced blood flow, infections, oxidative stress and chronic inflammation. Plasma-activated water (PAW) is emerging [...] Read more.
Diabetic foot ulcers have an enormous impact on patients’ quality of life and represent a major economic burden. The cause is delayed and incomplete wound healing due to hyperglycemia, reduced blood flow, infections, oxidative stress and chronic inflammation. Plasma-activated water (PAW) is emerging as a new therapeutic approach in wound treatment, as it has many of the advantages of cold atmospheric plasma but is easier to apply, thus allowing for widespread use. The aim of this study was to investigate the potential of PAW to improve wound healing in diabetic rats, with a focus on uncovering the underlying mechanisms. Two full-thickness wounds in control and diabetic animals were treated with PAW, and healing was monitored for 15 days at five time points. PAW improved wound healing in diabetic rats and mainly affected the inflammatory phase of wound healing. Application of PAW decreased the number of inflammatory cells, myeloperoxidase (MPO) and N-acetyl-b-D-glycosaminidase (NAG) activity, as well as the mRNA expression of proinflammatory genes in diabetic rats. Ten days after injury, PAW treatment increased collagen deposition in the diabetic animals by almost 10% without affecting collagen mRNA expression, and this is in correlation with a decrease in the Mmp-9/Timp-1 ratio. In conclusion, PAW treatment affects wound healing by reducing the inflammatory response and influencing extracellular matrix turnover, suggesting that it has great potential to accelerate the healing of diabetic wounds. Full article
(This article belongs to the Section Molecular Endocrinology and Metabolism)
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<p><b>The effect of PAW treatment on the progression of wound healing in control and diabetic rats</b>. (<b>A</b>) Blood glucose levels during the treatment period. (<b>B</b>) Representative photographs of the full-thickness skin wounds with or without PAW treatment in control and diabetic rats. (<b>C</b>) The wound closure area (%) during the healing process in control and diabetic rats with or without PAW treatment. C–Control non-diabetic group (<span class="html-italic">n</span> = 3); CP–PAW-treated control non-diabetic group (<span class="html-italic">n</span> = 3); D–diabetic group (<span class="html-italic">n</span> = 3); DP–PAW-treated diabetic group (<span class="html-italic">n</span> = 3). Values are expressed as means ± SEM, and the results of the ordinary two-way ANOVA and Tukey’s post hoc tests are presented in the tables below the graphs: dark grey <span class="html-italic">p</span> ≤ 0.01; blue <span class="html-italic">p</span> ≤ 0.001.</p>
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<p><b>The effect of PAW treatment on the progression of wound healing in control and diabetic rats</b>. (<b>A</b>) Blood glucose levels during the treatment period. (<b>B</b>) Representative photographs of the full-thickness skin wounds with or without PAW treatment in control and diabetic rats. (<b>C</b>) The wound closure area (%) during the healing process in control and diabetic rats with or without PAW treatment. C–Control non-diabetic group (<span class="html-italic">n</span> = 3); CP–PAW-treated control non-diabetic group (<span class="html-italic">n</span> = 3); D–diabetic group (<span class="html-italic">n</span> = 3); DP–PAW-treated diabetic group (<span class="html-italic">n</span> = 3). Values are expressed as means ± SEM, and the results of the ordinary two-way ANOVA and Tukey’s post hoc tests are presented in the tables below the graphs: dark grey <span class="html-italic">p</span> ≤ 0.01; blue <span class="html-italic">p</span> ≤ 0.001.</p>
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<p><b>Histological evaluation of wound healing in control and diabetic rats with or without PAW treatment</b>. (<b>A</b>) Haematoxylin–eosin staining of wounds with or without PAW treatment in control and diabetic rats. The progression of re-epithelialization at day VI is marked by yellow arrows (magnifications 2.5× and 10×), while the structure of granulation tissue is presented on day X after the injury (magnification 10×). (<b>B</b>) A schematic presentation of histologic measurements and formulas used for the calculation of epidermal closure (<b>C</b>), dermal closure (<b>D</b>) and total wound closure (<b>E</b>). C–Control non-diabetic group (<span class="html-italic">n</span> = 3); CP–PAW-treated control non-diabetic group (<span class="html-italic">n</span> = 3); D–diabetic group (<span class="html-italic">n</span> = 3); DP–PAW-treated diabetic group (<span class="html-italic">n</span> = 3). Values are expressed as means ± SEM, and the results of the ordinary two-way ANOVA and Tukey’s post hoc tests are presented in the tables below the graphs: light grey <span class="html-italic">p</span> ≤ 0.05; dark grey <span class="html-italic">p</span> ≤ 0.01; blue <span class="html-italic">p</span> ≤ 0.001.</p>
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<p><b>Histological evaluation of wound healing in control and diabetic rats with or without PAW treatment</b>. (<b>A</b>) Haematoxylin–eosin staining of wounds with or without PAW treatment in control and diabetic rats. The progression of re-epithelialization at day VI is marked by yellow arrows (magnifications 2.5× and 10×), while the structure of granulation tissue is presented on day X after the injury (magnification 10×). (<b>B</b>) A schematic presentation of histologic measurements and formulas used for the calculation of epidermal closure (<b>C</b>), dermal closure (<b>D</b>) and total wound closure (<b>E</b>). C–Control non-diabetic group (<span class="html-italic">n</span> = 3); CP–PAW-treated control non-diabetic group (<span class="html-italic">n</span> = 3); D–diabetic group (<span class="html-italic">n</span> = 3); DP–PAW-treated diabetic group (<span class="html-italic">n</span> = 3). Values are expressed as means ± SEM, and the results of the ordinary two-way ANOVA and Tukey’s post hoc tests are presented in the tables below the graphs: light grey <span class="html-italic">p</span> ≤ 0.05; dark grey <span class="html-italic">p</span> ≤ 0.01; blue <span class="html-italic">p</span> ≤ 0.001.</p>
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<p><b>The effect of PAW treatment on inflammatory response during wound healing in control and diabetic rats</b>. (<b>A</b>) Haematoxylin–eosin staining of granulation tissue of wounds with or without PAW treatment in control and diabetic rats at day VI (magnification 40×). Inflammatory cells are indicated by yellow and fibroblasts by green arrows. (<b>B</b>) The number of inflammatory cells counted per area. (<b>C</b>) Biochemical quantification of myeloperoxidase (MPO) activity as a measure of neutrophil infiltrate. (<b>D</b>) Biochemical quantification of N-acetylglucosaminidase (NAG) as a measure of macrophage infiltration. C–Control non-diabetic group (<span class="html-italic">n</span> = 3); CP–PAW-treated control non-diabetic group (<span class="html-italic">n</span> = 3); D–diabetic group (<span class="html-italic">n</span> = 3); DP–PAW-treated diabetic group (<span class="html-italic">n</span> = 3). Values are expressed as means ± SEM, and the results of the ordinary two-way ANOVA and Tukey’s post hoc tests are presented in the tables below the graphs: light grey <span class="html-italic">p</span> ≤ 0.05; dark grey <span class="html-italic">p</span> ≤ 0.01; blue <span class="html-italic">p</span> ≤ 0.001.</p>
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<p><b>The effect of PAW treatment on inflammatory response during wound healing in control and diabetic rats</b>. (<b>A</b>) Haematoxylin–eosin staining of granulation tissue of wounds with or without PAW treatment in control and diabetic rats at day VI (magnification 40×). Inflammatory cells are indicated by yellow and fibroblasts by green arrows. (<b>B</b>) The number of inflammatory cells counted per area. (<b>C</b>) Biochemical quantification of myeloperoxidase (MPO) activity as a measure of neutrophil infiltrate. (<b>D</b>) Biochemical quantification of N-acetylglucosaminidase (NAG) as a measure of macrophage infiltration. C–Control non-diabetic group (<span class="html-italic">n</span> = 3); CP–PAW-treated control non-diabetic group (<span class="html-italic">n</span> = 3); D–diabetic group (<span class="html-italic">n</span> = 3); DP–PAW-treated diabetic group (<span class="html-italic">n</span> = 3). Values are expressed as means ± SEM, and the results of the ordinary two-way ANOVA and Tukey’s post hoc tests are presented in the tables below the graphs: light grey <span class="html-italic">p</span> ≤ 0.05; dark grey <span class="html-italic">p</span> ≤ 0.01; blue <span class="html-italic">p</span> ≤ 0.001.</p>
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<p><b>The effect of PAW treatment on collagen deposition during wound healing in control and diabetic rats</b>. (<b>A</b>) Masson’s trichrome staining of wounds with or without PAW treatment in control and diabetic rats at days VI and X (magnifications 5× and 20×). (<b>B</b>) ImageJ software quantification of collagen (blue staining) expressed as % area within the sections. C–Control non-diabetic group (<span class="html-italic">n</span> = 3); CP–PAW-treated control non-diabetic group (<span class="html-italic">n</span> = 3); D–diabetic group (<span class="html-italic">n</span> = 3); DP–PAW-treated diabetic group (<span class="html-italic">n</span> = 3). Values are expressed as means of % area ± SEM, and the results of the ordinary two-way ANOVA and Tukey’s post hoc tests are presented in the tables below the graphs: light grey <span class="html-italic">p</span> ≤ 0.05; dark grey <span class="html-italic">p</span> ≤ 0.01; blue <span class="html-italic">p</span> ≤ 0.001.</p>
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<p>The effect of PAW treatment on the mRNA expression profile of selected genes in wound tissue from control and diabetic rats. C–Control non-diabetic group (<span class="html-italic">n</span> = 3); CP–PAW-treated control non-diabetic group (<span class="html-italic">n</span> = 3); D–diabetic group (<span class="html-italic">n</span> = 3); DP–PAW-treated diabetic group (<span class="html-italic">n</span> = 3). mRNA levels are presented relative to <span class="html-italic">Tbp</span> and are expressed as means ± SEM. The results of the ordinary two-way ANOVA and Tukey’s post hoc tests are presented in the tables below the graphs: light grey <span class="html-italic">p</span> ≤ 0.05; dark grey <span class="html-italic">p</span> ≤ 0.01; blue <span class="html-italic">p</span> ≤ 0.001.</p>
Full article ">Figure 5 Cont.
<p>The effect of PAW treatment on the mRNA expression profile of selected genes in wound tissue from control and diabetic rats. C–Control non-diabetic group (<span class="html-italic">n</span> = 3); CP–PAW-treated control non-diabetic group (<span class="html-italic">n</span> = 3); D–diabetic group (<span class="html-italic">n</span> = 3); DP–PAW-treated diabetic group (<span class="html-italic">n</span> = 3). mRNA levels are presented relative to <span class="html-italic">Tbp</span> and are expressed as means ± SEM. The results of the ordinary two-way ANOVA and Tukey’s post hoc tests are presented in the tables below the graphs: light grey <span class="html-italic">p</span> ≤ 0.05; dark grey <span class="html-italic">p</span> ≤ 0.01; blue <span class="html-italic">p</span> ≤ 0.001.</p>
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<p>Schematic diagram of the DBD-type atmospheric pressure plasma jet system used for water treatments.</p>
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16 pages, 839 KiB  
Article
Monitoring of Cadmium, Lead, and Mercury Levels in Seafood Products: A Ten-Year Analysis
by Luisa Garofalo, Marcello Sala, Claudia Focardi, Patrizio Pasqualetti, Daniela Delfino, Francesca D’Onofrio, Barbara Droghei, Francesca Pasquali, Valentina Nicolini, Flavia Silvia Galli, Paola Scaramozzino, Alessandro Ubaldi, Katia Russo and Bruno Neri
Foods 2025, 14(3), 451; https://doi.org/10.3390/foods14030451 - 30 Jan 2025
Viewed by 696
Abstract
The European Food Safety Authority (EFSA) monitors the presence and concentration of contaminants in food to mitigate health risks. EU legislation sets maximum levels of heavy metals in foods, including cadmium (Cd), lead (Pb), and total Hg (THg) in seafood, due to their [...] Read more.
The European Food Safety Authority (EFSA) monitors the presence and concentration of contaminants in food to mitigate health risks. EU legislation sets maximum levels of heavy metals in foods, including cadmium (Cd), lead (Pb), and total Hg (THg) in seafood, due to their toxicity. In the framework of official control, between 2014 and 2023, 5854 seafood samples were collected and 4300 analyses for THg, 3338 for Cd, and 2171 for Pb were performed using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and cold vapor atomic absorption spectrometry (CVAAS). The aim was to assess the proportion of contaminated foods in the dataset, the concentration of contaminants, and the potential health risks associated with their intake. Of the total samples analyzed, 142 (2.43%) were found to be non-compliant (n.c.). Concentrations exceeding the limits for Cd were primarily detected in cephalopods (n = 17), mainly squids. In contrast, Hg levels exceeded the limits in marine fish (n = 118), notably in swordfish (11.30% of n.c. samples among those analyzed for this species), sharks (6.48%), and tuna species (3.11%). Regarding Pb, only a single bivalve sample was found to exceed the maximum limits. A preliminary assessment of weekly exposure to Hg through swordfish consumption raised concerns about the frequent intake of marine top predators, particularly for vulnerable people. Full article
(This article belongs to the Special Issue Recent Advances in the Detection of Food Contaminants and Pollutants)
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Figure 1
<p>Number of chemical analyses conducted for the three heavy metals across various taxonomic groups of seafood. The number of samples analyzed for each taxonomic group is indicated in parentheses within the legend.</p>
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<p>Number of chemical analyses conducted over different years, categorized by the type of sampling plan.</p>
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<p>Percentage distribution of the three outcomes for the n = 9809 chemical analyses carried out for each heavy metal. Outcomes include the following: below the limit of detection (LOD), below the limit of quantification (LOQ), and measurable values. Note: LODs and LOQs were 0.006 mg/kg and 0.020 mg/kg for Cd and Pb and 0.010 mg/kg and 0.030 mg/kg for Hg, respectively.</p>
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<p>Box plots of (<b>a</b>) [Cd] (mg/kg) values of non-compliant samples of the two main cephalopods groups (squid and flying squid). Data for octopus and cuttlefish are not represented. (<b>b</b>) Total [Hg] (mg/kg) values of non-compliant samples for each group of marine fish species (swordfish: n = 59; shark species: n = 16; tuna species: n = 14; others with EU limit of 0.5 mg/kg: n = 26). Data from other species (n = 3) with a [Hg] limit of 1.0 mg/kg are not plotted. The median value is represented by a horizontal line, while X indicates the mean value.</p>
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14 pages, 3605 KiB  
Article
Adjunctive Treatment Effect of Non-Thermal Atmospheric Pressure Plasma in Periodontitis-Induced Rats
by Hee-Young Choi, Hyun-Joo Kim, Ju-Youn Lee and Ji-Young Joo
J. Clin. Med. 2025, 14(3), 896; https://doi.org/10.3390/jcm14030896 - 29 Jan 2025
Viewed by 449
Abstract
Background/Objectives: As non-thermal atmospheric pressure plasma (NTP) is known to have advantages in application in the medical field, we consider its applicability to periodontitis, a representative chronic inflammatory disease. The purpose of this study was to evaluate the effect of NTP in [...] Read more.
Background/Objectives: As non-thermal atmospheric pressure plasma (NTP) is known to have advantages in application in the medical field, we consider its applicability to periodontitis, a representative chronic inflammatory disease. The purpose of this study was to evaluate the effect of NTP in inhibiting the progression of periodontitis in a rat model when additionally used in scaling and root planing (SRP). Methods: To induce experimental periodontitis in 20 rats, ligatures were placed in the maxillary second molar and lipopolysaccharide from Porphyromonas gingivalis was injected around the teeth. Then, NTP treatment was performed for 2 or 5 min, together with scaling and root planing (SRP). To evaluate alveolar bone loss, micro-computed tomography (micro-CT) analysis and hematoxylin–eosin (H-E) staining were performed. Tartrate-resistant acid phosphatase (TRAP) analysis was performed to compare the number of osteoclasts, while immunohistochemistry (IHC) analysis was performed to determine the expression levels of receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG). Enzyme-linked immunosorbent assay (ELISA) analysis was performed for the detection of cytokines (TNF-α, IL-1β, and IL-10) in tissues and sera. Results: When SRP was combined with NTP, alveolar bone loss was decreased, the number of osteoclasts and RANKL expression were decreased, OPG expression was increased, and pro-inflammatory cytokine (TNF-α and IL-1β) levels were significantly decreased. Compared with the NTP treatment for 2 min, when treated for 5 min, less alveolar bone loss, fewer osteoclasts, a lower RANKL expression level, and a higher OPG expression level were observed. Conclusions: This study evaluated the adjunctive treatment effect of NTP in periodontitis-induced rats. Based on the results of this study, we suggest that supplemental NTP treatment may be a good option for non-surgical periodontal treatment; however, further studies are needed to elucidate the mechanism through which NTP suppresses periodontal inflammation. Full article
(This article belongs to the Section Dentistry, Oral Surgery and Oral Medicine)
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Figure 1
<p>Flowchart showing the experimental design. LPS injection and NTP treatment were performed once every 3 days. LPS: lipopolysaccharide; SRP: scaling and root planing; NTP: non-thermal atmospheric pressure plasma.</p>
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<p>Effect of NTP on alveolar bone resorption. Representative micro-CT images of (<b>A</b>) C group, (<b>B</b>) L group, (<b>C</b>) LR group, (<b>D</b>) LRP2 group, and (<b>E</b>) LRP5 group; (<b>F</b>) bar graph showing bone volume per total volume for the 5 groups. Red dashed box: ROI. NTP: non-thermal atmospheric pressure plasma. BV/TV: bone volume/total volume. Data represent mean ± SD (N = 4/group). No statistically significant differences among groups.</p>
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<p>Effect of NTP on alveolar bone resorption. Representative H-E staining images of (<b>A</b>) C group, (<b>B</b>) L group, (<b>C</b>) LR group, (<b>D</b>) LRP2 group, and (<b>E</b>) LRP5 group; (<b>F</b>) distance between CEJ and ABC. NTP: non-thermal atmospheric pressure plasma; PL: periodontal ligament; D: dentin; AB: alveolar bone; CEJ: cemento-enamel junction; ABC: alveolar bone crest. Scale bars = 50 μm. Data represent mean ± SD (N = 4/group). No statistically significant differences among groups.</p>
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<p>Effect of NTP on osteoclast formation. Representative TRAP staining images of (<b>A</b>) C group, (<b>B</b>) L group, (<b>C</b>) LR group, (<b>D</b>) LRP2 group, and (<b>E</b>) LRP5 group; (<b>F</b>) quantification of osteoclast number. NTP: non-thermal atmospheric pressure plasma; AB: alveolar bone; D: dentin; black arrows: multinucleated osteoclasts. Scale bars = 50 μm. Data represent mean ± SD (N = 4/group). *: statistically significant difference (<span class="html-italic">p</span> &lt; 0.01).</p>
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<p>Effect of NTP on the expression of RANKL and OPG. Representative IHC analysis images of (<b>A</b>) C group, (<b>B</b>) L group, (<b>C</b>) LR group, (<b>D</b>) LRP2 group, and (<b>E</b>) LRP5 group on RANKL expression; (<b>F</b>) IHC-stained area (%) on RANKL expression; IHC analysis images of (<b>G</b>) C group, (<b>H</b>) L group, (<b>I</b>) LR group, (<b>J</b>) LRP2 group, and (<b>K</b>) LRP5 group on OPG expression; (<b>L</b>) IHC-stained area (%); (<b>M</b>) RANKL/OPG ratio. NTP: non-thermal atmospheric pressure plasma. Scale bars = 20 μm. Data represent mean ± SD (N = 4/group). *: statistically significant difference (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of NTP on the expression of inflammatory/immune-related cytokines. (<b>A</b>–<b>C</b>) Detection amount of cytokine from tissue; (<b>D</b>–<b>F</b>) detection amount of cytokine from serum. (<b>A</b>,<b>D</b>) TNF-α, (<b>B</b>,<b>E</b>) IL-1β, and (<b>C</b>,<b>F</b>) IL-10. NTP: non-thermal atmospheric pressure plasma. Data represent mean ± SD (N = 4/group). *: statistically significant difference (<span class="html-italic">p</span> &lt; 0.05). **: statistically significant difference (<span class="html-italic">p</span> &lt; 0.01).</p>
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