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17 pages, 6152 KiB  
Article
Loss of CHOP Prevents Joint Degeneration and Pain in a Mouse Model of Pseudoachondroplasia
by Jacqueline T. Hecht, Alka C. Veerisetty, Mohammad G. Hossain, Debabrata Patra, Michele Carrer, Frankie Chiu, Dorde Relic, Paymaan Jafar-nejad and Karen L. Posey
Int. J. Mol. Sci. 2025, 26(1), 16; https://doi.org/10.3390/ijms26010016 - 24 Dec 2024
Abstract
Pseudoachondroplasia (PSACH), a severe dwarfing condition characterized by impaired skeletal growth and early joint degeneration, results from mutations in cartilage oligomeric matrix protein (COMP). These mutations disrupt normal protein folding, leading to the accumulation of misfolded COMP in chondrocytes. The MT-COMP mouse is [...] Read more.
Pseudoachondroplasia (PSACH), a severe dwarfing condition characterized by impaired skeletal growth and early joint degeneration, results from mutations in cartilage oligomeric matrix protein (COMP). These mutations disrupt normal protein folding, leading to the accumulation of misfolded COMP in chondrocytes. The MT-COMP mouse is a murine model of PSACH that expresses D469del human COMP in response to doxycycline and replicates the PSACH chondrocyte and clinical pathology. The basis for the mutant-COMP pathology involves endoplasmic reticulum (ER) stress signaling through the PERK/eIF2α/CHOP pathway. C/EBP homologous protein (CHOP), in conjunction with a TNFα inflammatory process, upregulates mTORC1, hindering autophagy clearance of mutant COMP protein. Life-long joint pain/degeneration diminishes quality of life, and treatments other than joint replacements are urgently needed. To assess whether molecules that reduce CHOP activity should be considered as a potential treatment for PSACH, we evaluated MT-COMP mice with 50% CHOP (MT-COMP/CHOP+/−), antisense oligonucleotide (ASO)-mediated CHOP knockdown, and complete CHOP ablation (MT-COMP/CHOP−/−). While earlier studies demonstrated that loss of CHOP in MT-COMP mice reduced intracellular retention, inflammation, and growth plate chondrocyte death, we now show that it did not normalize limb growth. ASO treatment reduced CHOP mRNA by approximately 60%, as measured by RT-qPCR, but did not improve limb length similar to MT-COMP/CHOP+/−. Interestingly, both 50% genetic reduction and complete loss of CHOP alleviated pain, while total ablation of CHOP in MT-COMP mice was necessary to preserve joint health. These results indicate that (1) CHOP reduction therapy is not an effective strategy for improving limb length and (2) pain and chondrocyte pathology are more responsive to intervention than the prevention of joint damage. Full article
(This article belongs to the Special Issue Advances in Molecular Research of Cartilage: 2nd Edition)
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Figure 1

Figure 1
<p>Effect of loss or reduction of CHOP on limb length and growth plate chondrocytes at 4 weeks. Femurs were collected at 4 to 5 weeks of age. (<b>A</b>) Femur lengths were measured from μCT images and growth plate widths were measured from H&amp;E images. Each group was compared to the age-matched MT-COMP control group. Femoral length in MT-COMP was not improved in the absence of or in diminished CHOP (MT-COMP/CHOP<sup>−/+</sup> and CHOP ASO-treated MT-COMP) but trended towards improvement in the absence of CHOP (MT-COMP/CHOP<sup>−/−</sup>). ASO-treated samples are separated by a vertical line to indicate that ASO-mediated knockdown of CHOP operates through a distinct mechanism compared to the genetic reduction of CHOP levels. Femur lengths were measured in at least 5 male mice, compared using a <span class="html-italic">t</span>-test. (<b>B</b>–<b>G</b>) H&amp;E staining of control (C57BL\6), MT-COMP, MT-COMP/CHOP<sup>−/+</sup> (50% CHOP), MT-COMP/CHOP<sup>−/−</sup> (CHOP absent), CHOP<sup>−/−</sup>, and CHOP ASO-treated MT-COMP growth plates at 4 weeks is shown. (<b>H</b>–<b>M</b>) P-eIF2α immunostaining of growth plates (brown signal) is shown in the lower panel. Representative growth plates are shown from the examination of at least 8 mice of both sexes. Bar = 100 μm * = <span class="html-italic">p</span> &lt; 0.05; *** = <span class="html-italic">p</span> &lt; 0.0005.</p>
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<p>Loss or reduction of CHOP reduces ER retention of mutant COMP. Growth plates from 4-week-old control (C57BL\6), MT-COMP, MT-COMP/CHOP<sup>−/+</sup> MT-COMP/CHOP<sup>−/−</sup>, CHOP<sup>−/−</sup>, and CHOP ASO-treated MT-COMP were immunostained for human-COMP (<b>A</b>–<b>F</b>), IL-6 (<b>G</b>–<b>L</b>), pS6 (<b>M</b>–<b>R</b>), PCNA (<b>S</b>–<b>X</b>) antibodies (brown signal), and apoptosis via terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate–biotin nick-end labeling (TUNEL) (<b>Y</b>–<b>AD</b>) (TUNEL green signal; nuclei are blue). ASO-treated samples are separated by a vertical line to indicate that ASO-mediated knockdown of CHOP operates through a distinct mechanism compared to the genetic reduction of CHOP levels. The human-COMP antibody specifically recognizes human mutant-COMP expressed in MT-COMP mice in response to DOX. Controls (<b>A</b>) and CHOP<sup>−/−</sup> (<b>E</b>) showed no intracellular staining for mutant-COMP compared to untreated MT-COMP growth plate chondrocytes where it was present (<b>B</b>). Representative growth plates are shown from examining at least 8 mice in both sexes. Scale bar = 50 μm.</p>
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<p>MT-COMP/CHOP<sup>−/−</sup> mice do not exhibit joint degeneration at 20 weeks. Four joint health parameters were assessed in control (gray bars), MT-COMP (blue bars), MT-COMP/CHOP<sup>−/+</sup> (light green bars), and MT-COMP/CHOP<sup>−/−</sup> (dark green bars) joints: proteoglycan levels in femoral articular cartilage (<b>A</b>) and tibia articular cartilage (<b>B</b>), synovitis (<b>C</b>), and bone/cartilage damage (<b>D</b>). The sum of the scores for each group is shown in panel (<b>E</b>). These assessments were conducted on a minimum of 10 male mice per group. Statistical analysis was performed using the Kruskal–Wallis test with post-hoc Dunn Test with Holm-adjusted <span class="html-italic">p</span>-values; * indicates <span class="html-italic">p</span> &lt; 0.05; ** indicates <span class="html-italic">p</span> &lt; 0.005; <span class="html-italic">p</span>-values between 0.05–0.1 are listed. All groups were compared to MT-COMP.</p>
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<p>Pain is reduced with the absence or reduction of CHOP in MT-COMP mice. Grooming was used as a proxy for pain assessment by measuring the efficiency of fluorescent dye removal from the fur. The higher score indicates better dye elimination (maximum score = 5). All male mice received DOX from birth until grooming was evaluated at 4, 8, 12, 16, 20, 24, 30, and 36 weeks in control C57BL\6 (Control) and MT-COMP, MT-COMP/CHOP<sup>−/+</sup>, and MT-COMP/CHOP<sup>−/−</sup> mice. All groups were compared to MT-COMP mice. The average grooming scores were analyzed with Kruskal–-Wallis with a post-hoc Dwass–Steel–Critchlow–Fligner (DSCF) significant pairwise test between the control and each group, and significant differences are shown by asterisks. For more information, refer to <a href="#ijms-26-00016-t001" class="html-table">Table 1</a>. These assessments were conducted on a minimum of 10 male mice per group. The standard deviation is shown by error bars. (Abbreviations: weeks = wks). ** <span class="html-italic">p</span> &lt; 0.05; *** <span class="html-italic">p</span> &lt; 0.0005.</p>
Full article ">Figure 5
<p>Articular cartilage chondrocyte ER stress is reduced with the absence or reduction of CHOP in MT-COMP mice. Articular cartilage from control (C57BL\6), MT-COMP, MT-COMP/CHOP<sup>−/+</sup> (reduced CHOP), and MT-COMP/CHOP<sup>−/−</sup> (absent CHOP) groups was immunostained for human-COMP (<b>A</b>–<b>E</b>), P-eIF2α (<b>F</b>–<b>J</b>), IL-6 (<b>K</b>–<b>O</b>), pS6 (<b>P</b>–<b>T</b>), p16INK4a (<b>U</b>–<b>Y</b>) (brown signal) and TUNEL (green signal with blue nuclei) (<b>Z</b>–<b>AD</b>) in 20-week-old mice. Representative growth plates from at least 8 mice (both sexes). Dotted line denotes the top of the articular cartilage; bar = 50 μm.</p>
Full article ">Figure 6
<p>Articular chondrocytes show dampened degradation in the absence or reduction of CHOP. (<b>A</b>) Schematic showing the interaction of molecules examined in articular cartilage from control (C57BL\6), MT-COMP, MT-COMP/CHOP<sup>−/+</sup> (reduced CHOP), MT-COMP/CHOP<sup>−/−</sup> (absent CHOP), and CHOP<sup>−/−</sup> mice were immunostained for IL-10 (<b>B</b>–<b>F</b>), SIRT1 (<b>G</b>–<b>K</b>), TNFα (<b>L</b>–<b>P</b>), and MMP13 (<b>Q</b>–<b>U</b>) antibodies at 20 weeks. Representative growth plates are shown from the examination of at least 8 mice (both sexes). Dotted line denotes the top of the articular cartilage; bar = 50 μm.</p>
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22 pages, 4731 KiB  
Article
Characterization and Molecular Insights of a Chromium-Reducing Bacterium Bacillus tropicus
by Shanjana Rahman Tuli, Md. Firoz Ali, Tabassum Binte Jamal, Md. Abu Sayem Khan, Nigar Fatima, Irfan Ahmed, Masuma Khatun and Shamima Akhtar Sharmin
Microorganisms 2024, 12(12), 2633; https://doi.org/10.3390/microorganisms12122633 - 19 Dec 2024
Viewed by 702
Abstract
Environmental pollution from metal toxicity is a widespread concern. Certain bacteria hold promise for bioremediation via the conversion of toxic chromium compounds into less harmful forms, promoting environmental cleanup. In this study, we report the isolation and detailed characterization of a highly chromium-tolerant [...] Read more.
Environmental pollution from metal toxicity is a widespread concern. Certain bacteria hold promise for bioremediation via the conversion of toxic chromium compounds into less harmful forms, promoting environmental cleanup. In this study, we report the isolation and detailed characterization of a highly chromium-tolerant bacterium, Bacillus tropicus CRB14. The isolate is capable of growing on 5000 mg/L Cr (VI) in an LB (Luria Bertani) agar plate while on 900 mg/L Cr (VI) in LB broth. It shows an 86.57% reduction ability in 96 h of culture. It can also tolerate high levels of As, Cd, Co, Fe, Zn, and Pb. The isolate also shows plant growth-promoting potential as demonstrated by a significant activity of nitrogen fixation, phosphate solubilization, IAA (indole acetic acid), and siderophore production. Whole-genome sequencing revealed that the isolate lacks Cr resistance genes in their plasmids and are located on its chromosome. The presence of the chrA gene points towards Cr(VI) transport, while the absence of ycnD suggests alternative reduction pathways. The genome harbors features like genomic islands and CRISPR-Cas systems, potentially aiding adaptation and defense. Analysis suggests robust metabolic pathways, potentially involved in Cr detoxification. Notably, genes for siderophore and NRP-metallophore production were identified. Whole-genome sequencing data also provides the basis for molecular validation of various genes. Findings from this study highlight the potential application of Bacillus tropicus CRB14 for bioremediation while plant growth promotion can be utilized as an added benefit. Full article
(This article belongs to the Special Issue Biotechnology for Environmental Remediation)
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Figure 1
<p>Graphical representation of the complete workflow for characterizing the chromium-reducing bacterium CRB14, starting with bacterial isolation, growth tolerance, and PGP analyses, and progressing through genome sequencing, functional annotation, and comparative analysis.</p>
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<p>Bar plot depicting the growth of isolate CRB14 at different concentrations of Cr. The cells were cultured on LB broth supplemented with 25, 50, 100 and 200, 300, 400, 500, 600, 700, 800, 900 mg/L Cr (VI). The optical density was measured after incubation for 24 h, 48 h, 72 h, and 96 h at 35 °C.</p>
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<p>Reduction in Cr (VI) by isolate CRB14. The cells were cultured in Luria Bertani broth supplemented with 0, 25, 50, 100, 200, 300, 400, 500, 600, 700, 800, and 900 mg/L Cr (VI). The Cr (VI) reduction activity was measured after incubation for 24 h, 48 h, 72 h, and 96 h at 35 °C.</p>
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<p>Bar plot representing the growth of isolate CRB14 in the presence of various heavy metals.</p>
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<p>Nuclear genome circle diagram of CRB14. From outside to inside, coding genes (positive-sense strand), coding genes (negative-sense strand), tRNA (blue) and rRNA (orange), tmRNA (black), CRISPR (green), Cas cluster (cyan), and GC ratio and GC-skew.</p>
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<p>A maximum-likelihood phylogenetic tree based on 16S rRNA gene sequences of <span class="html-italic">Bacillus tropicus</span> CRB14 and other closely related strains. The isolate of interest is highlighted in red.</p>
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<p>COG classifications of the genome. Each bar corresponds to a specific classification, highlighting these proteins’ diverse roles in metabolism and physiological processes. The abscissa represents the various COG categories, while the ordinate shows the number of genes assigned to each category.</p>
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<p>KEGG classification of the predicted coding sequences. The x-axis denotes the various pathways, and the y-axis indicates the number of genes assigned to each pathway. The bars are color coded according to the six major pathway classes, indicating that the majority of genes are involved in metabolism.</p>
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<p>GO functional classification of CRB14. The x-axis shows the GO categories, and the y-axis represents the −log10 (<span class="html-italic">p</span>-value) for the top 10 terms in biological process (blue), cellular component (yellow), and molecular function (green).</p>
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<p>Schematic diagram of nine secondary metabolite biosynthetic gene clusters in <span class="html-italic">B. tropicus</span> CRB14. Potential secondary metabolite biosynthetic gene clusters were predicted using antiSMASH. Color-coded blocks indicate different gene functions: dark red for core biosynthetic genes, light red for additional biosynthetic genes, blue for transport-related genes, green for regulatory genes, and gray for other genes.</p>
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13 pages, 3351 KiB  
Article
Identification and Characterization of Endophytic Fungus DJE2023 Isolated from Banana (Musa sp. cv. Dajiao) with Potential for Biocontrol of Banana Fusarium Wilt
by Longqi Jin, Rong Huang, Jia Zhang, Zifeng Li, Ruicheng Li, Yunfeng Li, Guanghui Kong, Pinggen Xi, Zide Jiang and Minhui Li
J. Fungi 2024, 10(12), 877; https://doi.org/10.3390/jof10120877 - 17 Dec 2024
Viewed by 363
Abstract
This study characterized an endophytic fungus, DJE2023, isolated from healthy banana sucker of the cultivar (cv.) Dajiao. Its potential as a biocontrol agent against banana Fusarium wilt was assessed, aiming to provide a novel candidate strain for the biological control of the devastating [...] Read more.
This study characterized an endophytic fungus, DJE2023, isolated from healthy banana sucker of the cultivar (cv.) Dajiao. Its potential as a biocontrol agent against banana Fusarium wilt was assessed, aiming to provide a novel candidate strain for the biological control of the devastating disease. The fungus was isolated using standard plant tissue separation techniques and fungal culture methods, followed by identification through morphological comparisons, multi-gene phylogenetic analyses, and molecular detection targeting Fusarium oxysporum f. sp. cubense (Foc) race 1 and race 4. Furthermore, assessments of its characteristics and antagonistic effects were conducted through pathogenicity tests, biological trait investigations, and dual-culture experiments. The results confirmed isolate DJE2023 to be a member of the Fusarium oxysporum species complex but distinct from Foc race 1 or race 4, exhibiting no pathogenicity to banana plantlets of cv. Fenza No.1 or tomato seedlings cv. money maker. Only minute and brown necrotic spots were observed at the rhizomes of banana plantlets of ‘Dajiao’ and ‘Baxijiao’ upon inoculation, contrasting markedly with the extensive necrosis induced by Foc tropical race 4 strain XJZ2 at those of banana cv Baxijiao. Notably, co-inoculation with DJE2023 and XJZ2 revealed a significantly reduced disease severity compared to inoculation with XJZ2 alone. An in vitro plate confrontation assay showed no significant antagonistic effects against Foc, indicating a suppressive effect rather than direct antagonism of DJE2023. Research on the biological characteristics of DJE2023 indicated lactose as the optimal carbon source for its growth, while maltose favored sporulation. The optimal growth temperature for this strain is 28 °C, and its spores can germinate effectively within the range of 25–45 °C and pH 4–10, demonstrating a strong alkali tolerance. Collectively, our findings suggest that DJE2023 exhibits weak or non-pathogenic properties and lacks direct antagonism against Foc, yet imparts a degree of resistance against banana Fusarium wilt. The detailed information provides valuable insight into the potential role of DJE2023 in integrated banana disease control, presenting a promising candidate for biocontrol against banana Fusarium wilt. Full article
(This article belongs to the Special Issue Fusarium spp.: A Trans-Kingdom Fungus)
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Figure 1
<p>Identification of endophytic fungus DJE2023 from banana cv. Dajiao. (<b>a</b>) The colony morphology of DJE2023. (<b>b</b>) Conidia. (<b>c</b>) Chlamydospores. (<b>d</b>) Multigene system phylogenetic tree. (<b>e</b>) Molecular identification using specific primers for <span class="html-italic">Fusarium oxysporum</span> f. sp. <span class="html-italic">cubense</span> (<span class="html-italic">Foc</span>) races 1 and 4. Among them, W106F/W106R are the universal primers for <span class="html-italic">F. oxysporum</span>, W1805F/W1805R are the specific primers for race 1of <span class="html-italic">Foc</span>, and W2987F/W2987R are the specific primers for race 4 of <span class="html-italic">Foc</span>. M: DL2000 Marker; 1,2: DJE-2023; 3: FOC Race 1; 4: FOC Race 4; 5: Water control.</p>
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<p>The pathogenicity test results for the banana endophytic strain DJE2023 and the <span class="html-italic">Fusarium oxysporum</span> f. sp. <span class="html-italic">cubense</span> (<span class="html-italic">Foc</span>) race 4 strain XJZ2. (<b>a</b>) The typical symptoms exhibited in the rhizomes of various banana cultivars. (<b>b</b>) The disease incidence statistics resulting from inoculating strain DJE2023 onto various banana cultivars. (<b>c</b>) The disease incidence statistics resulting from inoculating DJE2023 and XJZ2 onto banana cv. Baxijiao. The inoculation data of 30 plantlets were randomly divided into three groups for statistical analysis and the mean ± S.D. (n = 3). The significant difference (Duncan test, <span class="html-italic">p</span> &lt; 0.05) between two groups is represented by the superscript letters above the error bars.</p>
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<p>The biocontrol effect of DJE2023 on the banana plantlets cv. Baxijiao. (<b>a</b>) XJZ2, DJE2023, XJZ2, and DJE2023 co-infect the roots of banana plantlets; CK is water control. The disease conditions of leaves and roots are observed 45 days later. (<b>b</b>) Disease severity levels of banana roots; each treatment was replicated with 30 banana plantlets. (<b>c</b>) Disease index analyzed by diseased plantlet number of different disease grades. The inoculation data of 30 plantlets were randomly divided into three groups for statistical analysis and the mean ± S.D. (n = 3). The significant difference (Duncan test, <span class="html-italic">p</span> &lt; 0.05) between two groups is represented by the superscript letters above the error bars.</p>
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<p>Effects of carbon source on colony morphology, growth diameter, and sporulation of DJE2023. (<b>a</b>) The effect of carbon sources on the colony morphology of DJE2023. (<b>b</b>) The effect of carbon sources on the growth diameter of DJE2023. (<b>c</b>) The effect of carbon sources on the spore production of DJE2023. All data are presented as the mean ± standard deviation (n = 3). The significant difference (Duncan test, <span class="html-italic">p</span> &lt; 0.05) between two groups is represented by the superscript letters above the error bars.</p>
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<p>Effect of temperature on the morphology and growth diameter of DJE2023 colonies. (<b>a</b>) Colony morphology of DJE2023 under different temperatures. (<b>b</b>) Effect of temperature on the growth diameter of DJE2023. All data are presented as the mean ± standard deviation (n = 3). The significant difference (Duncan test, <span class="html-italic">p</span> &lt; 0.05) between two groups is represented by the superscript letters above the error bars.</p>
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<p>Effect of pH on colony morphology and growth diameter of DJE2023. (<b>a</b>) Colony morphology of DJE2023 under different pH levels. (<b>b</b>) Effect of pH on the growth diameter of DJE2023. All data are presented as the mean ± standard deviation (n = 3). The significant difference (Duncan test, <span class="html-italic">p</span> &lt; 0.05) between two groups is represented by the superscript letters above the error bars.</p>
Full article ">
14 pages, 2051 KiB  
Article
Facultatively Anaerobic Staphylococci Enable Anaerobic Cutibacterium Species to Grow and Form Biofilms Under Aerobic Conditions
by Jeffrey B. Kaplan, Michael Assa, Noor Mruwat, Miloslav Sailer, Suresh Regmi and Khalaf Kridin
Microorganisms 2024, 12(12), 2601; https://doi.org/10.3390/microorganisms12122601 - 16 Dec 2024
Viewed by 459
Abstract
Facultatively anaerobic Staphylococcus spp. and anaerobic Cutibacterium spp. are among the most prominent bacteria on human skin. Although skin microbes generally grow as multispecies biofilms, few studies have investigated the interaction between staphylococci and Cutibacterium spp. in dual-species biofilms. Here, we measured the [...] Read more.
Facultatively anaerobic Staphylococcus spp. and anaerobic Cutibacterium spp. are among the most prominent bacteria on human skin. Although skin microbes generally grow as multispecies biofilms, few studies have investigated the interaction between staphylococci and Cutibacterium spp. in dual-species biofilms. Here, we measured the mono- and dual-species biofilm formation of four staphylococcal species (S. epidermidis, S. hominis, S. capitis, and S. aureus) and two Cutibacterium spp. (C. acnes and C. avidum) cultured in vitro under both aerobic and anaerobic conditions. The biofilms were quantitated by rinsing them to remove planktonic cells, detaching the biofilm bacteria via sonication, and enumerating the cells by dilution plating. When cultured alone, staphylococci formed biofilms under both aerobic and anaerobic conditions, whereas Cutibacterium spp. formed biofilms only under anaerobic conditions. In co-culture, staphylococcal biofilm formation was unaffected by the presence of Cutibacterium spp., regardless of oxygen availability. However, Cutibacterium spp. biofilm formation was significantly enhanced in the presence of staphylococci, enabling robust growth under both anaerobic and aerobic conditions. Fluorescence confocal microscopy of the aerobic dual-species biofilms suggested that staphylococci create anaerobic niches at the base of the biofilm where C. acnes can grow. These findings demonstrate that staphylococci facilitate the colonization of Cutibacterium spp. in oxygen-rich environments, potentially explaining their presence in high numbers on the oxygen-exposed stratum corneum. Full article
(This article belongs to the Collection Feature Paper in Biofilm)
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Figure 1
<p><span class="html-italic">Cutibacterium acnes</span> and <span class="html-italic">Staphylococcus</span> spp. mono- and dual-species biofilms grown in glass tubes under aerobic and anaerobic conditions. (<b>a</b>) Biofilm formation by <span class="html-italic">S. epidermidis</span> strain 5 (<b>left panel</b>) and <span class="html-italic">C. acnes</span> strain HL086PA1 (<b>right panel</b>) cultured alone (mono-culture—black solid and dashed lines) or together (co-culture—red and blue lines) under aerobic and anaerobic conditions. After 72 h, tubes were rinsed to remove planktonic cells, biofilms were detached by sonication, and species were individually enumerated by dilution plating on selective agar. Graphs show mean CFU/tube values for triplicate tubes. Error bars indicate sd. (<b>b</b>) Biofilm formation by <span class="html-italic">C. acnes</span> strain HL086PA1 cultured alone (dashed line) or co-cultured with <span class="html-italic">S. epidermidis</span> strain 5 or <span class="html-italic">S. aureus</span> strain JE2 (solid lines) for 40 h under aerobic conditions. Tubes were processed as in panel (<b>a</b>). Graph shows mean CFU/tube values for triplicate tubes. Error bars indicate sd. (<b>c</b>) Photographs of 72-h-old <span class="html-italic">S. epidermdis</span> strain 5/<span class="html-italic">C. acnes</span> strain HL086PA1 dual-species biofilms after treating them for 15 min with phosphate-buffered saline (PBS), PBS supplemented with dispersin B (80 µg/mL), or PBS supplemented with DNase I (100 µg/mL), and then staining them with crystal violet. Duplicate tubes for each treatment are shown.</p>
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<p>Growth of <span class="html-italic">Cutibacterium</span> spp. in 15 mL conical-bottom polypropylene tubes. Four <span class="html-italic">Cutibacterium</span> spp. (species and strain names indicated at right) were cultured alone anaerobically (solid black line), alone aerobically (dashed black line), or co-cultured aerobically with each staphylococcal species and strain indicated at top (red lines). Tubes were sonicated after 40 h of incubation, and sonicates were diluted and plated on agar plates selective for <span class="html-italic">C. acnes</span>. Graphs show mean <span class="html-italic">C. acnes</span> CFU/tube values for triplicate tubes. Error bars were omitted for clarity.</p>
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<p>Growth of <span class="html-italic">C. acnes</span> strain 2 co-cultured with <span class="html-italic">S. epidermidis</span> strain JK4 in polypropylene tubes under aerobic conditions. After 40 h, tubes were sonicated, and sonicates were diluted and plated on selective agar for <span class="html-italic">C. acnes</span> CFU enumeration. (<b>a</b>) Tubes were incubated under static conditions or under dynamic conditions in a roller apparatus. (<b>b</b>). Media were supplemented with 20 µg/mL erythromycin or no antibiotic. (<b>c</b>) <span class="html-italic">S. epidermidis</span> was heat killed prior to inoculation. Graphs show mean CFU/tube values for triplicate tubes and error bars indicate sd.</p>
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<p>Growth of <span class="html-italic">C. acnes</span> strain 2 co-cultured with <span class="html-italic">S. epidermidis</span> strain JK4 in polystyrene microtiter plate wells. (<b>a</b>) <span class="html-italic">C. acnes</span> strain 2 was cultured alone (dashed line) or co-cultured with <span class="html-italic">S. epidermidis</span> strain JK4 (solid line) in 96-well, non-tissue-culture-treated, round-bottom microtiter plates under aerobic conditions. After 48 h, wells were scraped with a pipette tip, and contents of wells were diluted and plated on selective agar for <span class="html-italic">C. acnes</span> CFU enumeration. Graphs show mean CFU/well values for triplicate wells, and error bars indicate sd. (<b>b</b>) Same as (<b>a</b>), except bacteria were cultured in 24-well, tissue-culture-treated microtiter plate wells. (<b>c</b>) Same as (<b>b</b>), except surfaces of wells were seeded with NHEK cell monolayers.</p>
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<p>Colony biofilm assay. (<b>a</b>) Colony biofilms were cultured on a 25 mm diameter, 0.45 µm pore-size nylon filter placed on the surface of a tryptic soy agar plate. (<b>b</b>) <span class="html-italic">C. acnes</span> strain 2 was inoculated alone or in combination with <span class="html-italic">S. epidermidis</span> strain JK4 (indicated at bottom) onto filters. Plates were incubated anaerobically or aerobically (indicated at top) for 48 h. Filters were then transferred to 5 mL of saline and sonicated. Sonicates were diluted and plated on selective agar to enumerate <span class="html-italic">C. acnes</span> CFUs. Graph shows mean <span class="html-italic">C. acnes</span> CFU/filter values from a total of six filters from three independent experiments for each condition. Error bars indicate sd. Horizontal dashed lines indicate average starting <span class="html-italic">C. acnes</span> inoculum per filter (±1 sd).</p>
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<p>Growth of <span class="html-italic">C. acnes</span> strain 2 co-cultured with <span class="html-italic">S. epidermidis</span> strain JK4 in 15 mL polypropylene tubes under aerobic conditions. Values show mean CFU/tube of each species for triplicate tubes at each time point. Error bars indicate sd.</p>
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<p>Cross-sections of <span class="html-italic">C. acnes</span> strain 2 and <span class="html-italic">S. epidermidis</span> strain 1457/pCM29-GFP biofilms grown on glass coverslips, stained with DAPI, and visualized by fluorescence confocal microscopy. (<b>a</b>) Anaerobic <span class="html-italic">C. acnes</span> mono-species biofilm. (<b>b</b>) Aerobic <span class="html-italic">S. epidermidis</span> mono-species biofilm. (<b>c</b>) Aerobic <span class="html-italic">C. acnes</span>/<span class="html-italic">S. epidermidis</span> dual-species biofilm. Biofilm thickness ≈ 20 µm. Green, GFP; blue, DAPI.</p>
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17 pages, 3081 KiB  
Article
The Induction of Disease Resistance by Scopolamine and the Application of Datura Extract Against Potato (Solanum tuberosum L.) Late Blight
by Zhiming Zhu, Shicheng Liu, Yi Liu, Xinze Zhang, Zhiwen Shi, Shuting Liu, Zhenglin Zhu and Pan Dong
Int. J. Mol. Sci. 2024, 25(24), 13442; https://doi.org/10.3390/ijms252413442 - 15 Dec 2024
Viewed by 429
Abstract
Late blight, caused by Phytophthora infestans, is a devastating disease of potato. Our previous work illustrated that scopolamine, the main bioactive substance of Datura extract, exerts direct inhibitory effects on P. infestans, but it is unclear whether scopolamine and Datura extract [...] Read more.
Late blight, caused by Phytophthora infestans, is a devastating disease of potato. Our previous work illustrated that scopolamine, the main bioactive substance of Datura extract, exerts direct inhibitory effects on P. infestans, but it is unclear whether scopolamine and Datura extract can boost resistance to late blight in potato. In this study, P. infestans is used to infect scopolamine-treated potato pieces and leaves, as well as whole potatoes. We found that scopolamine-treated potato is resistant to P. infestans both in vitro and in vivo. The treatment of 4.5 g/L scopolamine reduces the lesion size of whole potato to 54% compared with the control after 20 d of the infection of P. infestans. The disease-resistant substance detection based on the kit method shows that scopolamine triggers the upregulation of polyphenoloxidase, peroxidase, superoxide dismutase activities, and H2O2 contents in potato tubers, and the decline of phenylalanine ammonia lyase and catalase activity. A total of 1682 significantly differentially expressed genes were detected with or without scopolamine treatment through high-throughput transcriptome sequencing and the DESeq2 software (version 1.24.0), including 705 upregulated and 977 downregulated genes. Scopolamine may affect the genes functioning in the cell wall, membrane and the plant-pathogen interaction. The addition of Datura extract could directly inhibit the mycelial growth of P. infestans on rye plate medium. In addition, P. infestans was found to be resistant to late blight in potato pieces treated with Datura extract. Datura extract can also be utilized in combination with the chemical fungicide Infinito in field experiments to lessen late blight symptoms and enhance potato yield. To our knowledge, this is the first study to detect the induction of disease resistance by scopolamine, and it also explores the feasibility of Datura extract in potato disease resistance. Full article
(This article belongs to the Special Issue Biocontrol of Plant Diseases and Insect Pests)
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<p>Scopolamine induces potatoes to resist late blight. Late blight symptoms of ‘Favorita’ potato leaves (<b>A</b>) and pieces (<b>B</b>) pretreated with different concentrations of scopolamine. (<b>C</b>) Late blight symptoms of ‘Qingshu 9’, ‘Xisen 6’, and ‘Hongmei’ potato pieces pretreated with scopolamine. (<b>D</b>) Symptoms of late blight in whole ‘Marco’ potatoes with or without scopolamine treatment. Proportion of lesion size of ‘Favorita’ potato leaves (<b>E</b>), pieces (<b>F</b>), and whole ‘Marco’ potatoes (<b>G</b>). d: days post inoculation with <span class="html-italic">P. infestans</span>. Tukey’s multiple comparisons test * <span class="html-italic">p</span> &lt; 0.0332, ** <span class="html-italic">p</span> &lt; 0.0021, *** <span class="html-italic">p</span> &lt; 0.0002, **** <span class="html-italic">p</span> &lt; 0.0001. 3 replicates per group.</p>
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<p>Change of disease-resistant substances in potato tubers with or without scopolamine treatment. (<b>A</b>) Phenylalanine ammonia (PAL) activity, (<b>B</b>) polyphenols oxidase (PPO) activity, (<b>C</b>) peroxidase (POD) activity, (<b>D</b>) superoxide dismutase (SOD) activity, (<b>E</b>) catalase (CAT) activity, (<b>F</b>) H<sub>2</sub>O<sub>2</sub> content in control group and scopolamine-treated group. h: hours after scopolamine treatment. d: days post-inoculation with <span class="html-italic">P. infestans</span>. Three replicates per group. Values represent the means ± standard error of 3 independent samples (Tukey’s multiple comparisons test, * <span class="html-italic">p</span> &lt; 0.0332, ** <span class="html-italic">p</span> &lt; 0.0021, *** <span class="html-italic">p</span> &lt; 0.0002).</p>
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<p>Main information of transcriptome sequencing in potato tubers under scopolamine treatment. (<b>A</b>) Distribution of differentially expressed genes. (<b>B</b>) qRT-PCR result. (<b>C</b>) GO annotation analysis diagram. The <span class="html-italic">X</span>–axis represents the number of genes compared to the secondary classification. (<b>D</b>) GO enrichment analysis. The horizontal axis represents the ratio of sample number of genes enriched in the rich factor (GO term) to the background number of annotated gene, and the color of the dot corresponds to different <span class="html-italic">p</span>-adjust ranges. (<b>E</b>) Histogram of KEGG. The <span class="html-italic">X</span>–axis is the number of genes annotated to the pathway. (<b>F</b>) KEGG enrichment analysis. The horizontal axis represents the ratio of rich factor (sample number of genes enriched in this pathway to background number of annotated genes). Three replicates per group.</p>
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<p><span class="html-italic">Datura</span> extract has dual effects to resist potato late blight. (<b>A</b>) Growth state of <span class="html-italic">Phytophthora infestans</span> on the medium supplemented with different concentrations of <span class="html-italic">Datura</span> extract. (<b>B</b>) The growth of <span class="html-italic">Phytophthora infestans</span> on the potato pieces pretreated with different concentrations of scopolamine. (<b>C</b>) The inhibition ratio of <span class="html-italic">Datura</span> extract against <span class="html-italic">Phytophthora infestans</span>. (<b>D</b>) The proportion of lesion size of potato pieces. Three replicates per group. Dunnett’s multiple comparisons test, * <span class="html-italic">p</span> &lt; 0.0332, ** <span class="html-italic">p</span> &lt; 0.0021, *** <span class="html-italic">p</span> &lt; 0.0002.</p>
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<p><span class="html-italic">Datura</span> extract could control late blight of potato and increase potato yield. (<b>A</b>) Late blight status of potato leaves at harvest. (<b>B</b>) Harvested potatoes. (The total length of the sign in the picture is 0.36 m) (<b>C</b>) Disease index of each treatment group. (The five surveys were given on 2, 6, 13, 19 April, and 5 May 2023). (<b>D</b>) Average yield of each treatment group. Treatment method: group 1. Control, treated with water; group 2. Infinito (1.5 mL); group 3. <span class="html-italic">Datura</span> extract (40 g); group 4. Infinito (0.15 mL); group 5. <span class="html-italic">Datura</span> extract (40 g) + Infinito (0.15 mL). (Tukey’s multiple comparisons test, * <span class="html-italic">p</span> &lt; 0.0332, ** <span class="html-italic">p</span> &lt; 0.0021.).</p>
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15 pages, 7940 KiB  
Article
Study on Fatigue Behavior and Fracture Mechanism of LMD Ti-6.5Al-3.5Mo-1.5Zr-0.3Si Alloy Based on Microstructure
by Yuxue Wu, Yongxin Wang, Yunmei Lu and Chenxi Zhao
Materials 2024, 17(24), 6112; https://doi.org/10.3390/ma17246112 - 13 Dec 2024
Viewed by 411
Abstract
This study explores the fatigue behavior and fracture mechanisms of TC11 titanium alloy formed by laser metal deposition (LMD) and subjected to double annealing. The research focuses on how the alloy’s unique microstructure, consisting of alternating equiaxed and columnar crystals, influences its fatigue [...] Read more.
This study explores the fatigue behavior and fracture mechanisms of TC11 titanium alloy formed by laser metal deposition (LMD) and subjected to double annealing. The research focuses on how the alloy’s unique microstructure, consisting of alternating equiaxed and columnar crystals, influences its fatigue performance. The microstructure’s basket-like α’ phase, made up of both plate-shaped and needle-like structures, leads to variations in crack growth behavior, as shown in the relationship between the crack growth rate and the stress intensity. An analysis of slip patterns reveals that equiaxed crystals undergo more frequent deformation, accelerating crack propagation compared to the more evenly distributed deformation in columnar crystals. These findings suggest a new approach for improving the fatigue resistance of 3D-printed titanium alloys by optimizing their microstructure. This study provides valuable insights for enhancing material toughness and extending the lifespan of titanium alloys in applications such as aerospace and biomedical engineering. Full article
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<p>Heat treatment process of deposited specimen.</p>
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<p>(<b>a</b>) Schematic illustration of the deposition direction. Schematic representation of specimens: (<b>b1</b>) Fatigue crack extension specimen, (<b>b2</b>) Fatigue tensile specimen.</p>
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<p>Fitting diagram of S-N curves in two different forming directions of Kt = 1 and Kt = 3.</p>
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<p>T1 sample crack source morphology (<b>a</b>); dimple morphology (<b>a1</b>); stable propagation zone morphology (<b>a2</b>); T2 sample crack source morphology (<b>b</b>); dimple morphology (<b>b1</b>); stable propagation zone morphology (<b>b2</b>).</p>
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<p>Schematic representation of the fatigue tensile fracture location (<b>a</b>); schematic representation of the wire-cut sampling (<b>b</b>); schematic representation of the macroscopic micro-morphology of the sample (<b>c</b>).</p>
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<p>Partial SEM diagrams of T1 (<b>a</b>) and T2 (<b>b</b>) crack propagation path.</p>
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<p>Columnar crystal region Euler diagram (<b>a</b>) and its corresponding orientation difference diagram (<b>c</b>) and equiaxed crystal region Euler diagram (<b>b</b>) and its corresponding orientation difference diagram(<b>d</b>).</p>
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<p>SEM diagrams of grain boundaries and secondary cracks along the crack propagation path. (a,b) Equiaxed and columnar grain boundary morphology (c) Secondary crack.</p>
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<p>Metallographic structure diagram of the boundary area between the equiaxed crystals and columnar crystals (<b>a</b>). 200 times metallographic structure diagram (<b>b</b>). Internal metallographic structure diagram of grains (<b>c</b>).</p>
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<p>Schematic diagram of fatigue crack growth path in L-T direction (<b>a</b>) and T-L direction (<b>b</b>).</p>
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<p>da/dN~ΔK diagram (<b>a</b>) and da/dN~a diagram (<b>b</b>) of L-T and T-L crack propagation specimens.</p>
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<p>Columnar intracrystalline IPF pattern (<b>a</b>). Polar pattern α (<b>a1</b>) β (<b>a2</b>) and equiaxed intracrystalline IPF pattern (<b>b</b>). Polar pattern α (<b>b1</b>) β (<b>b2</b>).</p>
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<p>SF statistics of columnar and equiaxed crystals.</p>
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<p>Columnar crystal phase distribution diagram (<b>a</b>). KAM part hcp crystal bcc crystal orientation diagram (<b>b</b>) and equiaxed crystal phase distribution diagram (<b>c</b>). KAM part hcp crystal bcc crystal orientation diagram (<b>d</b>).</p>
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15 pages, 1694 KiB  
Article
Trichoderma longibrachiatum (T6) Peptaibols Inhibiting the Monilia yunnanensis Growth and Inducing Pear Fruit Resistance in Its Infection
by Hang Lv, Shuwu Zhang, Nan Ma, Solomon Boamah and Bingliang Xu
Antioxidants 2024, 13(12), 1517; https://doi.org/10.3390/antiox13121517 - 12 Dec 2024
Viewed by 268
Abstract
Pear fruit brown rot, caused by Monilia yunnanensis, affects pear fruit yields and quality. The present study determined Trichoderma longibrachiatum T6 (T6) peptaibols as a biological control alternative to synthetic fungicides and assessed its efficacy against M. yunnanensis through dual plate culture [...] Read more.
Pear fruit brown rot, caused by Monilia yunnanensis, affects pear fruit yields and quality. The present study determined Trichoderma longibrachiatum T6 (T6) peptaibols as a biological control alternative to synthetic fungicides and assessed its efficacy against M. yunnanensis through dual plate culture and surface spraying at different concentrations. T6 peptaibols effectively inhibited M. yunnanensis growth, achieving an 85.99% inhibitory rate at 1250 µg/mL after inoculation on PDA medium for 5 days, and 84.57% control efficacy on pear fruit with the same concentration at 6 days. Treatment with T6 peptaibols significantly decreased the average contents of malondialdehyde (MDA) and hydrogen peroxide (H2O2), as well as electrolyte leakage, by 31.99%, 27.93%, and 21.00% from days 1 to 9 post-inoculation, respectively, in comparison to the negative control. Additionally, the average antioxidant enzyme activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD) and polyphenol oxidase (PPO) increased by 86.27%, 56.76%, 25.94%, and 47.88%, respectively; the average defense enzyme activities of phenylalanine ammonia-lyase (PAL), lipoxygenase (LOX), chitinase (CHI), and β-1,3-glucanase (β-Glu) increased by 63.00%, 55.70%, 26.19%, and 16.34%, respectively. Moreover, the expression levels of the antioxidant and defense-related genes (CAT, SOD, POD, PPO, CHI, LOX, PAL, β-Glu) were significantly upregulated by 2.80, 2.81, 3.03, 2.79, 3.37, 2.49, 2.73, and 1.83-folds at 3 days after inoculation compared to the negative control. Thus, T6 peptaibols effectively reduced the pathogen infection through growth inhibition and antioxidant defenses, thereby boosting fruit immunity. Full article
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<p>(<b>I</b>) Effect of T6 peptaibols on the expansional symptoms of brown rot lesions on pear fruits. (<b>A</b>) the lesion of healthy pear fruit treated with sterile water only; (<b>B</b>) the lesion of healthy pear fruit treated with 1% methanol only, (<b>C</b>) the lesion of healthy pear fruit treated with T6 peptaibols only, (<b>D</b>) the lesion of pear fruit treated with <span class="html-italic">M. yunnanensis</span> and sterile water (negative control), (<b>E</b>) the lesion of pear fruit treated with 1% methanol and <span class="html-italic">M. yunnanensis</span> (positive control), (<b>F</b>) the lesion of pear fruit treated with T6 peptaibols and <span class="html-italic">M. yunnanensis</span>, (<b>G</b>) cross-section of healthy pear fruit treated with water only, (<b>H</b>) cross-section of pear fruit treated with 1% methanol only, (<b>I</b>) cross-section of pear fruit treated with T6 peptaibols only, (<b>J</b>) cross-section of pear fruit treated with <span class="html-italic">M. yunnanensis</span> and sterile water (negative control), (<b>K</b>) cross-section of pear fruit treated with 1% methanol and <span class="html-italic">M. yunnanensis</span> (positive control), and (<b>L</b>) cross-section of pear fruit treated with T6 peptaibols and <span class="html-italic">M. yunnanensis</span>. (<b>II</b>) Effect of 1250 µg/mL concentration of T6 peptaibols on lesions expansion in pear fruits after inoculation with <span class="html-italic">M. yunnanensis</span>; Small bars represent the standard errors of the means. Different lowercase letters indicate significant differences at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of T6 peptaibols on (<b>A</b>) MDA content, (<b>B</b>) H<sub>2</sub>O<sub>2</sub> content and (<b>C</b>) Electrolyte leakage of pear fruits at different days. Small bars represent the standard errors of the means. Different lowercase letters indicate significant differences at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of T6 peptaibols treatment on (<b>A</b>) CAT, (<b>B</b>) SOD, (<b>C</b>) POD and (<b>D</b>) PPO antioxidant enzyme activities of pear fruits at different days. Small bars represent the standard errors of the means. Different lowercase letters indicate significant differences at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of T6 peptaibols treatment on (<b>A</b>) PAL, (<b>B</b>) LOX, (<b>C</b>) CHI and (<b>D</b>) β-Glu antioxidant enzyme activities of pear fruits at different days. Small bars represent the standard errors of the means. Different lowercase letters indicate significant differences at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of T6 peptaibols on the relative expression levels of (<b>A</b>) <span class="html-italic">CAT</span>, (<b>B</b>) <span class="html-italic">SOD</span>, (<b>C</b>) <span class="html-italic">POD</span> and (<b>D</b>) <span class="html-italic">PPO</span> antioxidants genes of pear fruits at day 3. Small bars represent the standard errors of the means. Different lowercase letters indicate significant differences at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of T6 peptaibols on the relative expression levels of (<b>A</b>) <span class="html-italic">PAL</span>, (<b>B</b>) <span class="html-italic">LOX</span>, (<b>C</b>) <span class="html-italic">CHI</span> and (<b>D</b>) <span class="html-italic">β-Glu</span> defense related genes of pear fruits at day 3. Small bars represent the standard errors of the means. Different lowercase letters indicate significant differences at <span class="html-italic">p</span> &lt; 0.05.</p>
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15 pages, 288 KiB  
Review
The Double-Edged Sword: Anterior Cruciate Ligament Reconstructions on Adolescent Patients—Growth Plate Surgical Challenges and Future Considerations
by Alexandria Mallinos and Kerwyn Jones
J. Clin. Med. 2024, 13(24), 7522; https://doi.org/10.3390/jcm13247522 - 11 Dec 2024
Viewed by 475
Abstract
The management of anterior cruciate ligament (ACL) injuries in pediatric patients presents unique challenges due to the presence of open growth plates in the proximal tibia and distal femur. Delaying ACL reconstruction until skeletal maturity may protect the physes but increases the risk [...] Read more.
The management of anterior cruciate ligament (ACL) injuries in pediatric patients presents unique challenges due to the presence of open growth plates in the proximal tibia and distal femur. Delaying ACL reconstruction until skeletal maturity may protect the physes but increases the risk of secondary injuries, such as meniscal tears and chondral damage, due to prolonged joint instability. Conversely, early surgical intervention restores knee stability but raises concerns about potential growth disturbances, including leg-length discrepancies and angular deformities. This narrative review examines current approaches to pediatric ACL management, highlighting the risks and benefits of both conservative and surgical treatments. Additionally, it explores the role of finite element modeling (FEM) as an innovative tool for pre-surgical planning. FEM offers a non-invasive method to optimize surgical techniques, minimize iatrogenic damage to growth plates, and improve patient outcomes. Despite its potential, FEM remains underutilized in clinical practice. This review underscores the need to integrate FEM into pediatric ACL care to enhance surgical precision, reduce complications, and improve long-term quality of life for young patients. By synthesizing available evidence, this review aims to provide clinicians with a comprehensive framework for decision-making and identify future directions for research in pediatric ACL reconstruction. Full article
9 pages, 1269 KiB  
Article
Correction of Femoral Torsional Deformities by Rotational Guided Growth
by Michael Zaidman, Naum Simanovsky, Vladimir Goldman and Eden Weisstub
J. Clin. Med. 2024, 13(24), 7514; https://doi.org/10.3390/jcm13247514 - 10 Dec 2024
Viewed by 318
Abstract
Background: Femoral torsional malalignment is a common cause of in-toeing and out-toeing in children, often leading to gait disturbances, functional limitations, and increased risk of falls. Traditionally, osteotomy was the only surgical option for correction. A minimally invasive technique known as rotational [...] Read more.
Background: Femoral torsional malalignment is a common cause of in-toeing and out-toeing in children, often leading to gait disturbances, functional limitations, and increased risk of falls. Traditionally, osteotomy was the only surgical option for correction. A minimally invasive technique known as rotational guided growth (RGG) has recently been introduced to address these malalignments. This study aims to assess the effectiveness of rotational femoral malalignment correction by rotational epiphysiodesis with tension band 8-plates (Orthofix, Verona, Italy). Methods: Eleven patients with in-toeing and out-toeing (19 femurs) were treated using RGG with 8-plates. The 8-plates were applied laterally and medially, with screws placed above and below the growth plate of the distal femur, angled obliquely to the long axis of the bone in opposite directions. Changes in foot progression angle (FPA), femoral version, the alteration in the angle between the 8-plates, and the rate of correction were recorded. Results: All patients reported functional gait improvement. The FPA was corrected from a mean of 32 degrees to 7 degrees, the femoral version improved from a mean of 60 degrees to 22 degrees. The angle between the 8-plates changed from a mean of 75 degrees to 28 degrees, with a correction rate of 4.1 degrees per month. The average time for correction was 11 months. No complications were observed during the treatment. Conclusions: RGG using 8-plates is a novel, minimally invasive surgical technique that effectively corrects rotational femoral deformities and may serve as a preferred alternative to derotational osteotomy in growing patients. Full article
(This article belongs to the Section Orthopedics)
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<p>Intraoperative radiographs of an 8-plate positioned obliquely for RGG, with (<b>A</b>) showing the lateral view and (<b>B</b>) showing the anteroposterior (AP) view.</p>
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<p>Radiographs showing the 8-plates in a horizontal orientation, with (<b>A</b>) representing the lateral view and (<b>B</b>) representing the anteroposterior (AP) view, indicating the end of their correction potential in the RGG process.</p>
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<p>Correction of FPA by RGG: On the <b>left</b> side, the patient exhibits noticeable in-toeing. On the <b>right</b> side, following RGG, the same patient demonstrates a corrected FPA, showing a successful improvement in gait alignment.</p>
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<p>(<b>A</b>,<b>B</b>) depict lateral radiographs of the knees of two different children, each showing an open growth plate and visible holes from previously placed (and now removed) metaphyseal screws. On follow-up, the holes in both cases have shifted further away from the growth plate, indicating that bone growth remains active in each child.</p>
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17 pages, 26738 KiB  
Article
Fatigue Crack Growth Performance of Q370qENH Weathering Bridge Steel and Butt Welds
by Yujie Yu, Xiang Zhang, Chunjian Hu, Liangkun Liu and Haibo Wang
Materials 2024, 17(23), 6015; https://doi.org/10.3390/ma17236015 - 9 Dec 2024
Viewed by 443
Abstract
Weathering steel possesses good atmospheric corrosion resistance and is increasingly applied in highway and railway bridges. The fatigue performance of the weld joint is an important issue in bridge engineering. This study experimentally investigates the microstructural properties and fracture crack growth behaviors of [...] Read more.
Weathering steel possesses good atmospheric corrosion resistance and is increasingly applied in highway and railway bridges. The fatigue performance of the weld joint is an important issue in bridge engineering. This study experimentally investigates the microstructural properties and fracture crack growth behaviors of a Q370qENH bridge weathering steel weld joint. The FCG parameters of the base steel, butt weld, and HAZs, considering the effect of different plate thicknesses and stress ratios, are analyzed. Microstructural features, microhardness, and fatigue fracture surfaces are carefully inspected. The FCG rates of different weld regions in the stable crack growth stage are obtained using integral formulas based on the Paris and Walker law. The test results indicate that the heating and cooling process during the welding of Q370qENH steel creates improved microstructures with refined grain sizes and fewer impurities, thus leading to improved FCG performances in the HAZ and weld regions. The crack growth rate of Q370qENH weld regions increases with the stress ratio, and the influencing extent increasingly ranks as the base steel, HAZ, and the weld. The thick plate has a slightly slower fatigue crack growth rate for the Q370qENH weld joints. The Q370qENH base steel presents the highest fatigue crack growth rate, followed by the heat-treated and HAZ cases, while the weld area exhibits the lowest FCG rate. The Paris law coefficients of different regions of Q370qENH welds are presented. The collected data serve as a valuable reference for future analyses of fatigue crack propagation problems of Q370qENH steel bridge joints. Full article
(This article belongs to the Special Issue Engineering Materials and Structural Integrity)
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<p>Design geometry and the practical product of butt welds.</p>
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<p>The optical microscope scan specimen and the construction.</p>
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<p>Metallographic structure: (<b>a</b>) weld joint region × 50 (Red spots indicate hardness measuring locations); (<b>b</b>) weld × 500; (<b>c</b>) CGHAZ × 500; (<b>d</b>) FGHAZ × 500; (<b>e</b>) base metal × 500.</p>
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<p>Metallographic structure: (<b>a</b>) HV-1000Z Vickers indenter; (<b>b</b>) microhardness results; (<b>c</b>) Vickers indenter imprint.</p>
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<p>CT specimen design: (<b>a</b>) specimen dimensions; (<b>b</b>) specimen figure; (<b>c</b>) specimen locations.</p>
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<p>Fatigue crack growth test setup: (<b>a</b>) test setup; (<b>b</b>) specimen setup; (<b>c</b>) DIC measurement.</p>
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<p>Fatigue crack growth curves: (<b>a</b>) crack propagation trajectory; (<b>b</b>) crack lengths of base metal; (<b>c</b>) crack lengths of HAZ; (<b>d</b>) crack lengths of weld.</p>
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<p>Comparison of FCG rates under different stress ratios: (<b>a</b>) base metal—8 mm thick; (<b>b</b>) base metal—10 mm thick; (<b>c</b>) weld—8 mm thick; (<b>d</b>) HAZ—8 mm thick.</p>
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<p>Comparison of FCG rates under different plate thicknesses: (<b>a</b>) base steel cases; (<b>b</b>) HAZ cases; (<b>c</b>) weld cases.</p>
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<p>Comparison of FCG rates under different stress ratios: (<b>a</b>) 0.1 stress ratio; (<b>b</b>) 0.2 stress ratio; (<b>c</b>) 0.5 stress ratio.</p>
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<p>Comparison of FCG rates between different CT groups: (<b>a</b>) CT specimens from weld joint; (<b>b</b>) CT specimens from heated plates.</p>
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<p>FCG performance comparisons between Q370qENH, 14MNNbq, and Q500D.</p>
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<p>Fracture morphology of 8H0.2-2.</p>
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<p>Fracture surface and microscope morphology of critical regions: (<b>a</b>) weld—8W0.1; (<b>b</b>) HAZ—8H0.1; (<b>c</b>) base steel—8B0.1.</p>
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<p>Fracture surface and microscope morphology of critical regions: (<b>a</b>) weld—8W0.1; (<b>b</b>) HAZ—8H0.1; (<b>c</b>) base steel—8B0.1.</p>
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<p>Fracture surface and microscope morphology of critical regions.</p>
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<p>Fracture surface of 6 mm and 8 mm thick 0.1 stress ratio cases.</p>
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29 pages, 4798 KiB  
Systematic Review
Lytic Spectra of Tailed Bacteriophages: A Systematic Review and Meta-Analysis
by Ivan M. Pchelin, Andrei V. Smolensky, Daniil V. Azarov and Artemiy E. Goncharov
Viruses 2024, 16(12), 1879; https://doi.org/10.3390/v16121879 - 4 Dec 2024
Viewed by 1057
Abstract
As natural predators of bacteria, tailed bacteriophages can be used in biocontrol applications, including antimicrobial therapy. Also, phage lysis is a detrimental factor in technological processes based on bacterial growth and metabolism. The spectrum of bacteria bacteriophages interact with is known as the [...] Read more.
As natural predators of bacteria, tailed bacteriophages can be used in biocontrol applications, including antimicrobial therapy. Also, phage lysis is a detrimental factor in technological processes based on bacterial growth and metabolism. The spectrum of bacteria bacteriophages interact with is known as the host range. Phage science produced a vast amount of host range data. However, there has been no attempt to analyse these data from the viewpoint of modern phage and bacterial taxonomy. Here, we performed a meta-analysis of spotting and plaquing host range data obtained on strains of production host species. The main metric of our study was the host range value calculated as a ratio of lysed strains to the number of tested bacterial strains. We found no boundary between narrow and broad host ranges in tailed phages taken as a whole. Family-level groups of strictly lytic bacteriophages had significantly different median plaquing host range values in the range from 0.18 (Drexlerviridae) to 0.70 (Herelleviridae). In Escherichia coli phages, broad host ranges were associated with decreased efficiency of plating. Bacteriophage morphology, genome size, and the number of tRNA-coding genes in phage genomes did not correlate with host range values. From the perspective of bacterial species, median plaquing host ranges varied from 0.04 in bacteriophages infecting Acinetobacter baumannii to 0.73 in Staphylococcus aureus phages. Taken together, our results imply that taxonomy of bacteriophages and their bacterial hosts can be predictive of intraspecies host ranges. Full article
(This article belongs to the Section Bacterial Viruses)
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<p>PRISMA flowchart of source identification and selection. PRISMA, preferred reporting items for systematic reviews and meta-analyses [<a href="#B33-viruses-16-01879" class="html-bibr">33</a>].</p>
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<p>Overview of the dataset: (<b>a</b>) bacterial host genera arranged by their prevalence (top 22 genera are shown); (<b>b</b>) distribution of bacteriophage genome size and morphology.</p>
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<p>Host range distributions are biased by published phage series and cannot be divided in parts. (<b>a</b>,<b>b</b>) The peaks in the original distributions are formed by data points originating from papers with high numbers of described bacteriophages, uncovering the prevalence of published series of bacteriophages with similar host ranges and a publication bias of large datasets. (<b>c</b>,<b>d</b>) The unbiased forms of the distributions were inferred by random choice of one data point from each paper in 500 replicates. The superimposed visualisations implied uniform distribution of spotting host ranges and a mixture of uniform and triangular distributions for plaquing host ranges. The visualisations are based on the host range data of all selected tailed bacteriophages, including the viruses with unknown taxonomic position within the class <span class="html-italic">Caudoviricetes</span>.</p>
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<p>Genetic diversity of bacteriophages in the sample visualised by phylogenetic network. The groups of phage genomes numbered from #1 to #4 were further analysed by phylogenetic tree construction (<a href="#app1-viruses-16-01879" class="html-app">Figures S1–S4</a>). Subfamily <span class="html-italic">Guernseyvirinae</span> was included in host range data analysis as a family-level group FLG-G. Host ranges of <span class="html-italic">Autographiviridae</span> members were analysed in two independent groups, FLG-A and FLG-AS (<span class="html-italic">Studiervirinae</span>).</p>
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<p>The distributions of bacteriophage host ranges differed between phage family-level groups: (<b>a</b>) Spotting host ranges. (<b>b</b>) Plaquing host ranges. Given the estimated error of repeated host range assessments at 0.1 (<a href="#app1-viruses-16-01879" class="html-app">Data S3</a>), PHR of <span class="html-italic">Straboviridae</span> and FLG-G, and the two host range types in <span class="html-italic">Drexlerviridae</span>, FLG-A and FLG-AS varied across the entire interval of values. (<b>c</b>) Differences between SHR value distributions and (<b>d</b>) differences between PHR value distributions assessed by Mann–Whitney U test. The order of family-level taxonomic groups of phages follows the increase in median spotting host ranges. med, median; <span class="html-italic">n</span>, number of host range data points; <span class="html-italic">nls</span>, number of literature sources; <span class="html-italic">S. thermophilus</span>, <span class="html-italic">Streptococcus thermophilus</span>.</p>
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<p>The distributions of bacteriophage host ranges differed between bacterial species: (<b>a</b>) Spotting host ranges. (<b>b</b>) Plaquing host ranges. Given the estimated error of repeated host range assessments at 0.1 (<a href="#app1-viruses-16-01879" class="html-app">Data S3</a>), SHR of <span class="html-italic">A. baumannii</span> phages, PHR of <span class="html-italic">E. coli</span> phages, and the two host range types of <span class="html-italic">S. enterica</span> phages varied across the entire interval of values. (<b>c</b>) Differences between SHR value distributions and (<b>d</b>) differences between PHR value distributions assessed by Mann–Whitney U test. The order of bacterial species follows the increase in median spotting host ranges. med, median; <span class="html-italic">n</span>, number of host range data points; <span class="html-italic">nls</span>, number of literature sources; <span class="html-italic">S. thermophilus</span>, <span class="html-italic">Streptococcus thermophilus</span>.</p>
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<p>In strictly lytic bacteriophages, broader plaquing host ranges may be associated with decreased proportion of efficiently utilised host strains. The lines depict linear regression analysis results with 95% confidence intervals shaded grey. In <span class="html-italic">E. coli</span> bacteriophages, there is a negative correlation between plaquing host ranges and the efficiency of plating (EOP). In the groups of bacteriophages propagating on <span class="html-italic">S. enterica</span> and all other hosts, the correlation cannot be seen.</p>
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<p>Bacteriophages morphotypes in the coordinates of host range and genome size: (<b>a</b>) Spotting host range. (<b>b</b>) Plaquing host range. There is no correlation between host ranges, morphotypes, and genome size. The visualisations are based on host range data of all selected tailed bacteriophages, including the viruses with unknown taxonomic position within the class <span class="html-italic">Caudoviricetes</span>.</p>
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<p>Sources of heterogeneity identified by factor analysis of mixed data: (<b>a</b>) Spotting host ranges. (<b>b</b>) Plaquing host ranges.</p>
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12 pages, 2767 KiB  
Article
The Impact of Using Laser and Milling Techniques to Create Zirconia Patterns on Streptococcus oralis Biofilm Formation
by Neusa Silva, Joana Marques, João Caramês, Filipe Silva, António Mata and Mariana Brito da Cruz
Ceramics 2024, 7(4), 1855-1866; https://doi.org/10.3390/ceramics7040116 - 3 Dec 2024
Viewed by 509
Abstract
This study aimed to evaluate zirconia dental implant surfaces patterned using Nd:YAG laser or conventional milling techniques against Streptococcus oralis adhesion and biofilm formation. Zirconia dental implant discs were subjected to surface patterning treatments and categorized into four groups: groove texturing by conventional [...] Read more.
This study aimed to evaluate zirconia dental implant surfaces patterned using Nd:YAG laser or conventional milling techniques against Streptococcus oralis adhesion and biofilm formation. Zirconia dental implant discs were subjected to surface patterning treatments and categorized into four groups: groove texturing by conventional milling (GM), pore texturing by conventional milling (PM), groove texturing by Nd:YAG laser (GL), and pore texturing by Nd: YAG laser (PL). Streptococcus oralis CECT 907T was cultivated on enriched blood agar plates and then transferred to a brain–heart infusion modified medium and incubated at 37 °C under anaerobic conditions until reaching the exponential growth phase. The bacterial suspension was then seeded on 24-well plates containing the treated discs. The viability of bacteria within the biofilm was determined based on colony-forming unit (CFU) counts, while the total biofilm was quantified by measuring its biomass. A qualitative analysis was conducted using scanning electron microscopy (SEM) images to evaluate the bacterial morphology. The statistical analysis of multigroup comparisons was performed using Kruskal–Wallis test with post hoc pairwise comparison, as well as Mann Whiney U test, with significance set at p < 0.05. After both 1 h and 24 h of incubation of Streptococcus oralis on the discs, all groups showed similar results, with no statistically significant differences (p > 0.05). A comparison of the Nd: YAG laser-treated surfaces with conventionally milled surfaces, as well as grooves versus pores for CFU counts, also revealed no statistically significant differences (p > 0.05) for both 1 h and 24 h of culture. Biomass quantification at both the 1 h and 24-h time points showed similar results across the groups, without statistical differences. When comparing the conventionally machined surfaces to Nd: YAG laser-treated surfaces in terms of biomass, no significant differences were observed (p > 0.05). Similarly, the comparison between groove-patterned surfaces and pore-patterned surfaces showed no statistically significant difference. The groove and pore patterns on zirconia surfaces with Nd: YAG laser or conventional milling did not change the Streptococcus oralis adhesion and biofilm formation behavior. Additional studies are recommended to expand our knowledge in this area. Full article
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<p>Schematic representation of zirconia disc production followed by randomization, texturing and patterning, sandblasting, and acid-etching. YTZP—yttria-stabilized zirconia powder; GM—groove patterns by conventional milling; PM—pore patterns by conventional milling; GL—groove patterns by laser Nd: YAG; PL—pore patterns by laser Nd: YAG; SBAE—sandblasted and acid-etched.</p>
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<p>Bar charts representing the mean CFU count per milliliter of <span class="html-italic">Streptococcus oralis</span>, presented as mean values, with error bars representing the standard deviation (n = 9). Group comparisons were performed using the Kruskal–Wallis test with post hoc pairwise comparisons. Statistical significance was set at <span class="html-italic">p</span> &lt; 0.05. GM—groove patterns by conventional milling; PM—pore patterns by conventional milling; GL—groove patterns by laser Nd: YAG; PL—pore patterns by laser Nd: YAG.</p>
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<p>Bar charts representing the CFU count per milliliter of <span class="html-italic">Streptococcus oralis</span> on conventional milling versus Nd:YAG laser zirconia-patterned surfaces, presented as mean values, with error bars indicating standard deviation (n = 18). Group comparisons were conducted using the Mann–Whitney U Test. Statistical significance was set at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Bar charts representing the CFU count per milliliter of <span class="html-italic">Streptococcus oralis</span> groove-patterned zirconia discs versus pore-patterned zirconia discs, presented as mean values, with error bars representing the standard deviation (n = 18). Group comparisons were conducted using the Mann–Whitney U Test. Statistical significance was set at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Bar charts illustrating the biomass quantification of <span class="html-italic">Streptococcus oralis</span>, presented as mean values, with error bars indicating standard deviation (n = 9). Group comparisons were conducted using the Kruskal–Wallis test with post hoc pairwise comparisons. Statistical significance was set at <span class="html-italic">p</span> &lt; 0.05. GM—groove patterns by conventional milling; PM—pore patterns by conventional milling; GL—groove patterns by laser Nd: YAG; PL—pore patterns by laser Nd: YAG.</p>
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<p>Bar charts depicting the biomass quantification of <span class="html-italic">Streptococcus oralis</span> on conventional milling versus Nd:YAG laser-patterned zirconia surfaces, presented as mean values, with error bars representing the standard deviation (n = 18). Group comparisons were conducted using the Mann–Whitney U Test. Statistical significance was set at <span class="html-italic">p</span> &lt;0.05.</p>
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<p>Bar charts depicting the biomass quantification of <span class="html-italic">Streptococcus oralis</span> on groove- versus pore-patterned zirconia surfaces, presented as mean values, with error bars representing the standard deviation (n = 18). Group comparisons were conducted using the Mann–Whitney U Test. Statistical significance was set at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>SEM images of <span class="html-italic">Streptococcus oralis</span> cultured on patterned zirconia after 1 h and 24 h incubations periods (n = 3). Images were acquired at 25 kV, ×5000 magnification, with scale bars representing 5 μm. GM—groove patterns by conventional milling; PM—pore patterns by conventional milling; GL—groove patterns by laser Nd: YAG; PL—pore patterns by laser Nd: YAG.</p>
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19 pages, 2022 KiB  
Article
Environmental and Economic LCA Comparison of Flexural Strengthening Solutions for a Reinforced Concrete Beam
by Pedro Frazão Pedroso, João R. Correia, José D. Silvestre, João P. Firmo and Mário Garrido
Materials 2024, 17(23), 5879; https://doi.org/10.3390/ma17235879 - 30 Nov 2024
Viewed by 431
Abstract
The construction sector is one of the largest creators and distributors of wealth, contributing to economic growth worldwide. However, this economic growth comes together with very high environmental impacts. Thus, rehabilitation solutions that can adapt the current building stock to today’s structural requirements [...] Read more.
The construction sector is one of the largest creators and distributors of wealth, contributing to economic growth worldwide. However, this economic growth comes together with very high environmental impacts. Thus, rehabilitation solutions that can adapt the current building stock to today’s structural requirements are needed, increasing structural safety, while avoiding the production of demolition waste and the extraction of virgin raw materials, hence lowering the construction sector’s environmental impacts. Such rehabilitation solutions need to be environmentally and economically sound so that stakeholders can make informed decisions based on their needs and priorities. This paper presents a case study of an existing reinforced concrete beam, whose flexural resistance is increased using four alternative strengthening solutions: concrete jacketing, without and with increasing the cross-section size, and plate bonding, using either carbon fibre-reinforced polymer (CFRP) strips or steel plates. These solutions are studied via an environmental and economic cradle-to-gate life cycle assessment (LCA), resulting in a comprehensive comparison of their environmental and economic impacts, followed by a multicriteria and sensitivity analysis and eco-cost approach to determine the optimal solution. According to the criteria considered in the study, when environmental impacts are more valued, the concrete jacketing solution presents the best results and, when cost is dominant in the decision, the bonding of CFRP strips becomes the optimal solution. Full article
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<p>Illustration of the different strengthening methods: (<b>a</b>) CJE; (<b>b</b>) CJI; (<b>c</b>) ASP; (<b>d</b>) ACF.</p>
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<p>Product stage and construction process (adapted from EN 15804:2012+A2:2019 [<a href="#B31-materials-17-05879" class="html-bibr">31</a>]).</p>
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<p>Method CJE: relative environmental impacts of each material.</p>
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<p>Method CJI: relative environmental impacts of each material.</p>
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<p>Method ASP: relative environmental impacts of each material.</p>
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<p>Method ACF: relative environmental impacts of each material.</p>
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18 pages, 2770 KiB  
Article
Seed Inoculation with Halotolerant Strains Enhance Brassicaceae Seedling Establishment Under Saline Conditions
by Carlos González-Cobo, Glòria Escolà, Roser Tolrà, Mercè Llugany, Charlotte Poschenrieder, Eliana Bianucci and Silvia Busoms
Agriculture 2024, 14(12), 2184; https://doi.org/10.3390/agriculture14122184 - 29 Nov 2024
Viewed by 351
Abstract
Soil salinity inhibits germination and seedling establishment, causing patchy crop stands, uneven growth, and poor yields. This study aims to evaluate the early-stage salinity tolerance of Brassicaceae seeds inoculated with plant growth-promoting bacterial (PGPB) strains (E1 and T7) isolated from saline soils. Non-inoculated [...] Read more.
Soil salinity inhibits germination and seedling establishment, causing patchy crop stands, uneven growth, and poor yields. This study aims to evaluate the early-stage salinity tolerance of Brassicaceae seeds inoculated with plant growth-promoting bacterial (PGPB) strains (E1 and T7) isolated from saline soils. Non-inoculated and inoculated seeds of Lobularia maritima, Sinapis alba, and Brassica napus were cultivated under control and salinity conditions, first in agar plates to assess a germination inhibitory concentration of salt for each species and later in soil irrigated with water containing 0 or 75 mM NaCl. Our results indicate that T7 was the only strain able to increase the germination of L. maritima under saline conditions. However, an increase in shoot biomass, root length, and number of branches was observed in L. maritima and S. alba plants inoculated with T7 and in B. napus with E1. Concomitantly, those seedlings exhibited less oxidative damage and greater capacity to balance plant reactive oxygen species production. This study suggests that inoculation of seeds with halotolerant PGPB strains is a suitable strategy for improving the negative effects of salinity in the early stages. Nonetheless, the observed specific plant–host interaction highlights the need for establishing tailored PGPB–crop associations for specific unfavourable environmental conditions. Full article
(This article belongs to the Section Seed Science and Technology)
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<p>Effects of salt on the germination of Brassicaceae species. Germination rate of <span class="html-italic">L. maritima</span> (maroon), <span class="html-italic">B. napus</span> (green), and <span class="html-italic">S. alba</span> (yellow) sown on MS plates with increasing concentrations of NaCl. Different letters indicate significant differences among NaCl treatments (Duncan test, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effects of PGPB seed inoculation on the germination of Brassicaceae species exposed to salinity. (<b>A</b>) Relative germination rate (Germination<sub>Salt</sub>/Germination<sub>Control</sub>) of <span class="html-italic">L. maritima</span>, <span class="html-italic">B. napus</span>, and <span class="html-italic">S. alba</span> seeds N/I (grey) or inoculated with T7 (yellow) or E1 (blue) sown in MS plates with 0 or 75 mM/150 mM NaCl. Different letters indicate significant differences among inoculation conditions (Duncan test, <span class="html-italic">p</span> &lt; 0.05). (<b>B</b>) Percentage of germinated seeds (light color) and established seedlings (dark color) of <span class="html-italic">L. maritima</span>, <span class="html-italic">B. napus</span>, and <span class="html-italic">S. alba</span> seeds N/I (grey) or inoculated with T7 (yellow) or E1 (blue) sown in sterile substrate irrigated with water containing 0 or 75 mM NaCl.</p>
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<p>Seedling growth of PGPB-inoculated Brassicaceae species exposed to salinity. Mean ± SE of shoot length (cm) of (<b>A</b>) <span class="html-italic">L. maritima</span>, (<b>B</b>) <span class="html-italic">B. napus</span>, and (<b>C</b>) <span class="html-italic">S. alba</span> seedlings N/I (grey) or inoculated with T7 (yellow) or E1 (blue) cultivated in sterile substrate irrigated with water containing 0 (solid lines) or 75 mM NaCl (dashed lines). Asterisks indicate significant differences among inoculation conditions (MANOVA repeated measures, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Root growth and architecture of PGPB-inoculated Brassicaceae species exposed to salinity. Mean ± SE of (<b>A</b>) root length (cm), (<b>B</b>) root volume (cm<sup>3</sup>), (<b>C</b>) number of root branches, and (<b>D</b>) number of root tips of <span class="html-italic">L. maritima</span>, <span class="html-italic">B. napus</span>, and <span class="html-italic">S. alba</span> seedlings N/I (grey) or inoculated with T7 (yellow) or E1 (blue) cultivated in sterile substrate irrigated with water containing 0 or 75 mM NaCl. Different letters indicate significant differences among inoculation conditions (Duncan test, <span class="html-italic">p</span> &lt; 0.05), while different numbers indicate significant differences among salt treatments (<span class="html-italic">t</span>-test, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Salinity tolerance of PGPB-inoculated Brassicaceae species. (<b>A</b>) Mean ± SE of aerial biomass (shoot dry weight, g), (<b>B</b>) representative pictures, (<b>C</b>) salinity tolerance index, and (<b>D</b>) leaf Na<sup>+</sup> concentration (mg/g DW) and Na:K ratio of <span class="html-italic">L. maritima</span>, <span class="html-italic">B. napus</span>, and <span class="html-italic">S. alba</span> seedlings N/I (grey) or inoculated with T7 (yellow) or E1 (blue) cultivated in sterile substrate irrigated with water containing 0 or 75 mM NaCl. Different letters indicate significant differences among inoculation conditions (Duncan test, <span class="html-italic">p</span> &lt; 0.05), while different numbers indicate significant differences among salt treatments (<span class="html-italic">t</span>-test, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Oxidative damage in roots of PGPB-inoculated Brassicaceae species exposed to salinity. Mean ± SE of (<b>A</b>) root hydrogen peroxide concentration (µmol H<sub>2</sub>O<sub>2</sub>/g FW), (<b>B</b>) root lipid peroxidation (µmol TBARs/g FW), and (<b>C</b>) FDA-PI-stained roots (10× magnification) of <span class="html-italic">L. maritima</span>, <span class="html-italic">B. napus</span>, and <span class="html-italic">S. alba</span> seedlings N/I (grey) or inoculated with T7 (yellow) or E1 (blue) cultivated in sterile substrate irrigated with water containing 0 or 75 mM NaCl. Different letters indicate significant differences among inoculation conditions (Duncan test, <span class="html-italic">p</span> &lt; 0.05), while different numbers indicate significant differences between salt treatments (<span class="html-italic">t</span>-test, <span class="html-italic">p</span> &lt; 0.05).</p>
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11 pages, 1823 KiB  
Article
High Revision Rate After Transphyseal ACL Reconstruction in Skeletally Immature Patients
by Benjamin Bartek, Tobias Jung, Theresa Lackner, Imke Schatka, Clemens Gwinner and Thula Walter-Rittel
J. Pers. Med. 2024, 14(12), 1129; https://doi.org/10.3390/jpm14121129 - 29 Nov 2024
Viewed by 376
Abstract
Objectives: There remains considerable debate regarding the optimal management of anterior cruciate ligament (ACL) injuries in skeletally immature patients. This study aims to evaluate the clinical outcomes of transphyseal ACL reconstruction in patients with open growth plates. Methods: This retrospective study included skeletally [...] Read more.
Objectives: There remains considerable debate regarding the optimal management of anterior cruciate ligament (ACL) injuries in skeletally immature patients. This study aims to evaluate the clinical outcomes of transphyseal ACL reconstruction in patients with open growth plates. Methods: This retrospective study included skeletally immature patients with full-thickness ACL tears and confirmed open physis. ACL reconstructions were performed using a four-strand semitendinosus autograft, with an additional gracilis tendon graft if needed. The surgical technique emphasized tibial and femoral physeal-sparing tunnel placement to minimize disruption of the growth plates. Clinical assessment included measurements for limb length discrepancy, knee stability, and growth disturbances. Functional outcomes were evaluated using IKDC 2000, Lysholm, and KOOS scores, while ligament stability was assessed with KT-1000 arthrometer measurements at routine follow-up. Results: A total of 31 consecutive patients (15 females, 16 males; mean age 13.6 ± 1.8 years, range 9–16 years) were included. Mean follow-up was 49 ± 26 months (range 18–93 months). The mean time to return to sports was 8.8 ± 4.4 months. Eight patients (26%) experienced ACL graft rupture and underwent revision ACL reconstruction. One additional patient required partial meniscectomy. The overall revision rate was 29%. The mean subjective IKDC score was 91.8 ± 7.2, with Lysholm and KOOS scores of 96.6 ± 7.9 and 94.2 ± 5.3, respectively. No significant growth disturbances were noted. The mean side-to-side difference in KT-1000 testing was 2.2 ± 1.5 mm. Patients who underwent revision ACL reconstruction showed significantly greater length growth compared with those with intact ACL reconstruction (p = 0.02). Spearman correlation revealed a significant association between length growth and anterior tibial translation (p = 0.02, r = 0.46). Conclusions: Transphyseal ACL reconstruction in skeletally immature patients provides favorable clinical and radiological outcomes, with minimal risk of growth disturbance. Most patients returned to pre-injury levels of athletic activity. However, the high revision rate emphasizes the complexity of managing ACL injuries in this population. Full article
(This article belongs to the Special Issue Personalized Medicine in Orthopaedics, 2nd Edition)
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<p>Example of a 3.0 Tesla knee MRI: (<b>A</b>) coronal PD tse fs, (<b>B</b>) sagittal T2 tse, and (<b>C</b>) conventional X-ray in AP view of 14-year-old male patient with open physis and acute ACL rupture. PD tse fs (proton density-weighted turbo spin echo sequence with fat saturation), T2 tse (T2-weighted turbo spin echo sequence), ap (anterior–posterior); large arrow is ACL rupture; star is bone marrow edema.</p>
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<p>Example of a postoperative 3.0 Tesla knee MRI: (<b>A</b>) sagittal and (<b>B</b>) coronal PD tse fs, and (<b>C</b>) sagittal PD tse of a 14-year-old female patient with open physis and postoperative MRI after ACL reconstruction. PD tse fs (proton density-weighted turbo spin echo sequence with fat saturation), PD tse (proton density-weighted turbo spin echo sequence).</p>
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<p>Visualization of the distribution of IKDC, Lysholm, and KOOS scores, showing predominantly good to very good results. While the IKDC score displays some variability, the Lysholm and KOOS scores are more consistently clustered near the upper range.</p>
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